Targeted chromosomal genomic alterations in plants using modified single stranded oligonucleotides

FIELD OF THE INVENTION

[0001] The technical field of the invention is oligonucleotide-directed repair or alteration of plant genetic information using novel chemically modified oligonucleotides.

BACKGROUND OF THE INVENTION

[0002] A number of methods have been developed specifically to alter the genomic information of plants. These methods generally include the use of vectors such as, for example, T-DNA, carrying nucleic acid sequences encoding partial or complete portions of a particular protein which is expressed in a cell or tissue to effect the alteration. The expression of the particular protein then results in the desired phenotype. See, for example, U.S. Pat. No. 4,459,355 which describes a method for transforming plants with a DNA vector and U.S. Pat. No. 5,188,642 which describes cloning or expression vectors containing a transgenic DNA sequence which when expressed in plants confers resistance to the herbicide glyphosate. The use of such transgene-containing vectors adds one or more exogenous copies of a gene in a usually random fashion at one or more integration sites of the plant's genome at some variable frequency. The introduced gene may be foreign or may be derived from the host plant. Any gene which was originally present in the genome, which may be, for example, a normal allelic variant, mutated, defective, and/or functional copy of the introduced gene, is retained in the genome of the host plant.

[0003] These methods of gene alteration are problematic in that complications which can compromise the vigor, productivity, yield, etc. of the plant may result. One such problem is that insertion of exogenous nucleic acid at random location(s) in the genome can have deleterious effects. The random nature of this insertion and/or the use of exogenous promoters can also cause the timing, location or strength of expression of the introduced transgene to be inappropriate or unpredictable. Another problem with such systems includes the addition of unnecessary and unwanted genetic material to the genome of the recipient, including, for example, T-DNA ends or other vector remnants, exogenous control sequences required to allow production of the transgene protein, which control sequences may be exogenous or native to the host plant and/or the transgene, and reporter genes or resistance markers. Such remnants and added sequences may have presently unrecognized consequences, for example, involving genetic rearrangements of the recipient genomes. In addition, concerns have been raised with consumption, especially by humans, of plants containing such exogenous genetic material.

[0004] More recently, simpler systems involving poly- or oligo-nucleotides have been described for use in the alteration of genomic DNA. These chimeric RNA-DNA oligonucleotides, requiring contiguous RNA and DNA bases in a double-stranded molecule folded by complementarity into a double hairpin conformation, have been shown to effect single basepair or frameshift alterations, for example, for mutation or repair of plant, animal or fungal genomes. See, for example, WO 99/07865 and U.S. Pat. No. 5,565,350. In the chimeric RNA-DNA oligonucleotide, an uninterrupted stretch of DNA bases within the molecule is required for sequence alteration of the targeted genome while the obligate RNA residues are involved in complex stability. Due to the length, backbone composition, and structural configuration of these chimeric RNA-DNA molecules, they are expensive to synthesize and difficult to purify. Moreover, if the RNA-containing strand of the chimeric RNA-DNA oligonucleotide is designed so as to direct gene alteration, a series of mutagenic reactions resulting in nonspecific base alteration can result. Such a result reduces the utility of such a molecule in methods designed for targeted gene alteration.

[0005] Alternatively, other oligo- or poly-nucleotides have been used which require a triplex forming, usually polypurine or polypyrimidine, structural domain which binds to a DNA helical duplex through Hoogsteen interactions between the major groove of the DNA duplex and the oligonucleotide. Such oligonucleotides may have an additional DNA reactive moiety, such as psoralen, covalently linked to the oligonucleotide. These reactive moieties function as effective intercalation agents, stabilize the formation of a triplex and can be mutagenic. Such agents may be required in order to stabilize the triplex forming domain of the oligonucleotide with the DNA double helix if the Hoogsteen interactions from the oligonucleotide/target base composition are insufficient. See, e.g., U.S. Pat. No. 5,422,251. The utility of these oligonucleotides for directing targeted gene alteration is compromised by a high frequency of nonspecific base changes.

[0006] In more recent work, the domain for altering a genome is linked or tethered to the triplex forming domain of the bi-functional oligonucleotide, adding an additional linking or tethering functional domain to the oligonucleotide. See, e.g., Culver et al., Nature Biotechnology 17: 989-93 (1999). Such chimeric or triplex forming molecules have distinct structural requirements for each of the different domains of the complete poly- or oligo-nucleotide in order to effect the desired genomic alteration in either episomal or chromosomal targets.

[0007] Other genes, e.g. CFTR, have been targeted by homologous recombination using duplex fragments having several hundred basepairs. See, e.g., Kunzelmann et al., Gene Ther. 3:859-867 (1996). Similar efforts to target genes by homologous recombination in plants using large fragments of DNA had some success. See Kempin et al., Nature 389:802-803 (1997). However, the efficiency and reproducibility of the published homologous recombination approach in plants has severely limited the widespread use of this method.

[0008] Earlier experiments to mutagenize an antibiotic resistance indicator gene by homologous recombination used an unmodified DNA oligonucleotide rather than larger fragments of DNA, wherein the oligonucleotide had no functional domains other than a region of complementary sequence to the target. See Campbell et al., New Biologist 1: 223-227 (1989). These experiments required large concentrations of the oligonucleotide, exhibited a very low frequency of episomal modification of a targeted exogenous plasmid gene not normally found in the cell and have not been reproduced. However, as shown in examples herein, we have observed that an unmodified DNA oligonucleotide can convert a base at low frequency which is detectable using the assay systems described herein.

[0009] Oligonucleotides designed for use in the targeted alteration of genetic information are significantly different from oligonucleotides designed for antisense approaches. For example, antisense oligonucleotides are perfectly complementary to and bind an mRNA strand in order to modify expression of a targeted mRNA and are used at high concentration. As a consequence, they are unable to produce a gene conversion event by either mutagenesis or repair of a defect in the chromosomal DNA of a host genome. Furthermore, the backbone chemical composition used in most oligonucleotides designed for use in antisense approaches renders them inactive as substrates for homologous pairing or mismatch repair enzymes and the high concentrations of oligonucleotide required for antisense applications can be toxic with some types of nucleotide modifications. In addition, antisense oligonucleotides must be complementary to the mRNA and therefore, may not be complementary to the other DNA strand or to genomic sequences that span the junction between intron sequence and exon sequence.

[0010] Artificial chromosomes can be useful for the screening purposes identified herein. These molecules are man-made linear or circular DNA molecules constructed from essential cis-acting DNA sequence elements that are responsible for the proper replication and partitioning of natural chromosomes (Murray et al., 1983). The essential elements are: (1) Autonomous Replication Sequences (ARS), (2) Centromeres, and (3) Telomeres.

[0011] Yeast artificial chromosomes (YACs) allow large segments of genomic DNA to be cloned and modified (Burke et al., Science 236:806; Peterson et al., Trends Genet. 13:61 (1997); Choi, et al., Nat. Genet., 4:117-223 (1993), Davies, et al., Biotechnology 11:911-914 (1993), Matsuura, et al., Hum. Mol. Genet., 5:451-459 (1996), Peterson et al., Proc. Natl. Acad. Sci., 93:6605-6609 (1996); and Schedl, et al., Cell, 86:71-82 (1996)). Other vectors also have been developed for the cloning of large segments of genomic DNA, including cosmids, and bacteriophage P1 (Sternberg et al., Proc. Natl. Acad. Sci. U.S.A., 87:103-107 (1990)). YACs have certain advantages over these alternative large capacity cloning vectors (Burke et al., Science, 236:806-812 (1987)). The maximum insert size is 35-30 kb for cosmids, and 100 kb for bacteriophage P1, both of which are much smaller than the maximal insert size for a YAC.

[0012] An alternative to YACs are cloning systems based on the E. coli fertility factor that have been developed to construct large genomic DNA insert libraries. They are bacterial artificial chromosomes (BACs) and P-1 derived artificial chromosomes (PACs) (Mejia et al., Genome Res. 7:179-186 (1997); Shizuya et al., Proc. Natl. Acad. Sci. 89:8794-8797 (1992); Ioannou et al., Nat. Genet., 6:84-89 (1994); Hosoda et al., Nucleic Acids Res. 18:3863 (1990)). BACs are based on the E. coli fertility plasmid (F factor); and PACs are based on the bacteriophage P1. These vectors propagate at a very low copy number (1-2 per cell) enabling genomic inserts up to 300 kb in size to be stably maintained in recombination deficient hosts. The PACs and BACs are circular DNA molecules that are readily isolated from the host genomic background by classical alkaline lysis (Birnboim et al., Nucleic Acids Res. 7:1513-1523 (1979)). In addition, BACs have been developed for transformation of plants with high-molecular weight DNA using the T-DNA system (Hamilton, Gene 24:107-116 (1997); Frary & Hamilton, Transgenic Res. 10: 121-132 (2001)).

[0013] A need exists for simple, inexpensive oligonucleotides capable of producing targeted alteration of genetic material such as those described herein as well as methods to identify optimal oligonucleotides that accurately and efficiently alter target DNA.

SUMMARY OF THE INVENTION

[0014] Novel, modified single-stranded nucleic acid molecules that direct gene alteration in plants are identified and the efficiency of alteration is analyzed both in vitro using a cell-free extract assay and in vivo using a yeast system and a plant system. The alteration in an oligonucleotide of the invention may comprise an insertion, deletion, substitution, as well as any combination of these. Site specific alteration of DNA is not only useful for studying function of proteins in vivo, but it is also useful for creating plants with desired phenotypes, including, for example, environmental stress tolerance, improved nutritional value, herbicide resistance, disease resistance, modified oil production, modified starch production, and altered floral morphology including selective sterility. As described herein, oligonucleotides of the invention target directed specific gene alterations in genomic double-stranded DNA in cells. The target genomic DNA can be nuclear chromosomal DNA as well as plastid or mitochondrial chromosomal DNA. The target DNA can also be a transgene present in the plant cell, including, for example, a previously introduced T-DNA. For screening purposes, the target plant DNA can also be extrachromosomal DNA present in plant or non-plant cells in various forms including, e.g., mammalian artificial chromosomes (MACs), PACs from P-1 vectors, yeast artificial chromosomes (YACs), bacterial artificial chromosomes (BACs), plant artificial chromosomes (PLACs), as well as episomal DNA, including episomal DNA from an exogenous source such as a plasmid or recombinant vector. Many of these artificial chromosome constructs containing plant DNA can be obtained from a variety of sources, including, e.g., the Arabidopsis Biological Resource Center (ABRC) at the Ohio State University, and the Rice Genome Research Program at the MAFF DNA bank in Ibaraki, Japan. The target DNA may be transcriptionally silent or active. In a preferred embodiment, the target DNA to be altered is the non-transcribed strand of a genomic DNA duplex. In a more preferred embodiment, the target DNA to be altered is the non-transcribed strand of a transcribed gene of a genomic DNA duplex.

[0015] The low efficiency of targeted gene alteration obtained using unmodified DNA oligonucleotides is believed to be largely the result of degradation by nucleases present in the reaction mixture or the target cell. Although different modifications are known to have different effects on the nuclease resistance of oligonucleotides or stability of duplexes formed by such oligonucleotides (see, e.g., Koshkin et al., J. Am. Chem. Soc., 120:13252-3), we have found that it is not possible to predict which of any particular known modification would be most useful for any given alteration event, including for the construction of gene alteration oligonucleotides, because of the interaction of different as yet unidentified proteins during the gene alteration event. Herein, a variety of nucleic acid analogs have been developed that increase the nuclease resistance of oligonucleotides that contain them, including, e.g., nucleotides containing phosphorothioate linkages or 2′-O-methyl analogs. We recently discovered that single-stranded DNA oligonucleotides modified to contain 2′-O-methyl RNA nucleotides or phosphorothioate linkages can enable specific alteration of genetic information at a higher level than either unmodified single-stranded DNA or a chimeric RNA/DNA molecule. See, for example, copending applications U.S. application Ser. No. 60/208,538, U.S. application Ser. No. 60/244,989, U.S. application Ser. No. 09/818,875, international application no. PCT/US01/09761 and Gamper et al., Nucleic Acids Research 28: 4332-4339 (2000), the disclosures of which are incorporated herein in their entirety by reference. We also found that additional nucleic acid analogs which increase the nuclease resistance of oligonucleotides that contain them, including, e.g., “locked nucleic acids” or “LNAs”, xylo-LNAs and L-ribo-LNAs; see, for example, Wengel & Nielsen, WO 99/14226; Wengel, WO 00/56748; Wengel, WO 00/66604; and Jakobsen & Koshkin, WO 01/25478 also allow specific targeted alteration of genetic information.

[0016] The assay allows for determining the optimum length of the oligonucleotide, optimum sequence of the oligonucleotide, optimum position of the mismatched base or bases, optimum chemical modification or modifications, optimum strand targeted for identifying and selecting the most efficient oligonucleotide for a particular gene alteration event by comparing to a control oligonucleotide. Control oligonucleotides may include a chimeric RNA-DNA double hairpin oligonucleotide directing the same gene alteration event, an oligonucleotide that matches its target completely, an oligonucleotide in which all linkages are phosphorothiolated, an oligonucleotide fully substituted with 2′-O-methyl analogs or an RNA oligonucleotide. Such control oligonucleotides either fail to direct a targeted alteration or do so at a lower efficiency as compared to the oligonucleotides of the invention. The assay further allows for determining the optimum position of a gene alteration event within an oligonucleotide, optimum concentration of the selected oligonucleotide for maximum alteration efficiency by systematically testing a range of concentrations, as well as optimization of either the source of cell extract by testing different plants or strains, or testing cells derived from different plants or strains, or plant cell lines. Using a series of single-stranded oligonucleotides, comprising all RNA or DNA residues and various mixtures of the two, several new structures are identified as viable molecules in nucleotide conversion to direct or repair a genomic mutagenic event. When extracts from mammalian, plant and fungal cells are used and are analyzed using a genetic readout assay in bacteria, single-stranded oligonucleotides having one of several modifications are found to be more active than a control RNA-DNA double hairpin chimera structure when evaluated using an in vitro gene repair assay. Similar results are also observed in vivo using yeast, mammalian and plant cells. Molecules containing various lengths of modified bases were found to possess greater activity than unmodified single-stranded DNA molecules.

DETAILED DESCRIPTION OF THE INVENTION

[0017] The present invention provides oligonucleotides having chemically modified, nuclease resistant residues, preferably at or near the termini of the oligonucleotides, and methods for their identification and use in targeted alteration of plant genetic material, including gene mutation, targeted gene repair and gene knockout. The oligonucleotides are preferably used for mismatch repair or alteration by changing at least one nucleic acid base, or for frameshift repair or alteration by addition or deletion of at least one nucleic acid base. The oligonucleotides of the invention direct any such alteration, including gene correction, gene repair or gene mutation and can be used, for example, to introduce a polymorphism or haplotype or to eliminate (“knockout”) a particular protein activity. For example, gene alterations that knockout a particular protein activity can be obtained using oligonucleotides designed to convert a codon in the coding region of the protein to a stop codon, thus prematurely terminating translation of the protein. Oligonucleotides that introduce stop codons in the open-reading-frame of the protein are one embodiment of the invention. Generally, oligonucleotides that introduce stop codons early in the open-reading-frame of the protein are preferred. If the open-reading-frame contains more than one methionine, oligonucleotides that introduce stop codons after the second methionine are preferred. Additionally, if the gene exhibits alternative splice sites, oligonucleotides that introduce stop codons in exons after the alternative splice site are preferred. The following table provides examples of codons that can be converted to stop codons by altering a single oligonucleotide. A skilled artisan could readily identify other codons that can be converted to stop codons by altering one, two or three of the base pairs in a given codon. Similarly, a skilled artisan could readily identify codons that can be converted to stop codons by a frameshift mutations that inserts or deletes one or two base pairs in the open-reading-frame. It is also understood that more than one stop codon can be generated in a single open-reading-frame and that these stop codons can be adjacent in the sequence or separated by intervening codons. Where more than one stop codon is introduced into a single open-reading-frame, such alterations can be generated by a single or multiple oligonucleotides and can be generated simultaneously or by sequential mutagenesis of the target nucleic acid.

1CorrespondingOriginal codons*stop codonGGA (glycine), AGA (arginine), CGA (arginine), TTATGA(leucine), TCA (serine), TGT (cysteine), TGG(tryptophan), TGC (cysteine)AAG (lysine), GAG (glutamate), CAG (glutamine), TTGTAG(leucine), TCG (serine), TGG (tryptophan), TAT(cysteine), TAC (tyrosine)AAA (lysine), GAA (glutamate), CAA (glutamine), TTATAA(leucine), TCA (serine), TAT (cysteine), TAC(tyrosine)*The amino acid encoded by the original codon is shown in parentheses and the base targeted for alteration to convert the codon to the corresponding stop codon is underlined and in bold

[0018] The oligonucleotides of the invention are designed as substrates for homologous pairing and repair enzymes and as such have a unique backbone composition that differs from chimeric RNA-DNA double hairpin oligonucleotides, antisense oligonucleotides, and/or other poly- or oligo-nucleotides used for altering genomic DNA, such as triplex forming oligonucleotides. The single-stranded oligo-nucleotides described herein are inexpensive to synthesize and easy to purify. In side-by-side comparisons, an optimized single-stranded oligonucleotide comprising modified residues as described herein is significantly more efficient than a chimeric RNA-DNA double hairpin oligonucleotide in directing a base substitution or frameshift mutation in a cell-free extract assay.

[0019] We have discovered that single-stranded oligonucleotides having a DNA domain surrounding the targeted base, with the domain preferably central to the poly- or oligo-nucleotide, and having at least one modified end, preferably at the 3′ terminal region, are able to alter a target genetic sequence and with an efficiency that is higher than chimeric RNA-DNA double hairpin oligonucleotides disclosed in U.S. Pat. No. 5,565,350. Preferred oligonucleotides of the invention have at least two modified bases on at least one of the termini, preferably the 3′ terminus of the oligonucleotide. Oligonucleotides of the invention can efficiently be used to introduce targeted alterations in a genetic sequence of DNA in the presence of human, animal, plant, fungal (including yeast) proteins and in cells of different types including, for example, plant cells, fungal cells including S. cerevisiae, Ustillago maydis, Candida albicans, and mammalian cells. Particularly preferred are cells and cell extracts derived from plants including, for example, experimental model plants such as Chiamydomonas reinhardtii, Physcomitrella patens, and Arabidopsis thaliana in addition to crop plants such as cauliflower (Brassica oleracea), artichoke (Cynara scolymus), fruits such as apples (Malus, e.g. domesticus), mangoes (Mangifera, e.g. indica), banana (Musa, e.g. acuminata), berries (such as currant, Ribes, e.g. rubrum), kiwifruit (Actinidia, e.g. chinensis), grapes (Vitis, e.g. vinifera), bell peppers (Capsicum, e.g. annuum), cherries (such as the sweet cherry, Prunus, e.g. avium), cucumber (Cucumis, e.g. sativus), melons (Cucumis, e.g. melo), nuts (such as walnut, Juglans, e.g. regia; peanut, Arachis hypogeae), orange (Citrus, e.g. maxima), peach (Prunus, e.g. persica), pear (Pyra, e.g. communis), plum (Prunus, e.g. domestica), strawberry (Fragaria, e.g. moschata or vesca), tomato (Lycopersicon, e.g. esculentum); leaves and forage, such as alfalfa (Medicago, e.g. sativa or truncatula), cabbage (e.g. Brassica oleracea), endive (Cichoreum, e.g. endivia), leek (Allium, e.g. porrum), lettuce (Lactuca, e.g. sativa), spinach (Spinacia, e.g. oleraceae), tobacco (Nicotiana, e.g. tabacum); roots, such as arrowroot (Maranta, e.g. arundinacea), beet (Beta, e.g. vulgaris), carrot (Daucus, e.g. carota), cassava (Manihot, e.g. esculenta), turnip (Brassica, e.g. rapa), radish (Raphanus, e.g. sativus), yam (Dioscorea, e.g. esculenta), sweet potato (Ipomoea batatas); seeds, including oilseeds, such as beans (Phaseolus, e.g. vulgaris), pea (Pisum, e.g. sativum), soybean (Glycine, e.g. max), cowpea (Vigna unguiculata), mothbean (Vigna aconitifolia), wheat (Triticum, e.g. aestivum), sorghum (Sorghum e.g. bicolor), barley (Hordeum, e.g. vulgare), corn (Zea, e.g. mays), rice (Oryza, e.g. sativa), rapeseed (Brassica napus), millet (Panicum sp.), sunflower (Helianthus annuus), oats (Avena sativa), chickpea (Cicer, e.g. arietinum); tubers, such as kohlrabi (Brassica, e.g. oleraceae), potato (Solanum, e.g. tuberosum) and the like; fiber and wood plants, such as flax (Linum e.g. usitatissimum), cotton (Gossypium e.g. hirsutum), pine (Pinus sp.), oak (Quercus sp.), eucalyptus (Eucalyptus sp.), and the like and ornamental plants such as turfgrass (Lolium, e.g. rigidum), petunia (Petunia, e.g. x hybrida), hyacinth (Hyacinthus orientalis), carnation (Dianthus e.g. caryophyllus), delphinium (Delphinium, e.g. ajacis), Job's tears (Coix lacryma-jobi), snapdragon (Antirrhinum majus), poppy (Papaver, e.g. nudicaule), lilac (Syringa, e.g. vulgaris), hydrangea (Hydrangea e.g. macrophylla), roses (including Gallicas, Albas, Damasks, Damask Perpetuals, Centifolias, Chinas, Teas and Hybrid Teas) and ornamental goldenrods (e.g. Solidago spp.). Such plant cells can then be used to regenerate whole plants according to methods described herein or any method known in the art. The DNA domain of the oligonucleotides is preferably fully complementary to one strand of the gene target, except for the mismatch base or bases responsible for the gene alteration event(s). On either side of the preferably central DNA domain, the contiguous bases may be either RNA bases or, preferably, are primarily DNA bases. The central DNA domain is generally at least 8 nucleotides in length. The base(s) targeted for alteration in the most preferred embodiments are at least about 8, 9 or 10 bases from one end of the oligonucleotide.

[0020] According to certain embodiments, one or both of the termini of the oligonucleotides of the present invention comprise phosphorothioate modifications, LNA backbone (including LNA derivatives and analogs) modifications, or 2′-O-methyl base analogs, or any combination of these modifications. Oligonucleotides comprising 2′-O-methyl or LNA analogs are a mixed DNA/RNA polymer. The oligonucleotides of the invention are, however, single-stranded and are not designed to form a stable internal duplex structure within the oligonucleotide. The efficiency of gene alteration is surprisingly increased with oligonucleotides having internal complementary sequence comprising phosphorothioate modified bases as compared to 2′-O-methyl modifications. This result indicates that specific chemical interactions are involved between the converting oligonucleotide and the proteins involved in the conversion. The effect of other such chemical interactions to produce nuclease resistant termini using modifications other than LNA (including LNA derivatives or analogs), phosphorothioate linkages, or 2′-O-methyl analog incorporation into an oligonucleotide can not yet be predicted because the proteins involved in the alteration process and their particular chemical interaction with the oligonucleotide substituents are not yet known and cannot be predicted.

[0021] In the examples, oligonucleotides of defined sequence are provided for alteration of genes in particular plants. Provided the teachings of the instant application, one of skill in the art could readily design oligonucleotides to introduce analogous alterations in homologous genes from any plant. Furthermore, in the tables of these examples, the oligonucleotides of the invention are not limited to the particular sequences disclosed. The oligonucleotides of the invention include extensions of the appropriate sequence of the longer 120 base oligonucleotides which can be added base by base to the smallest disclosed oligonucleotides of 17 bases. Thus the oligonucleotides of the invention include for each correcting change, oligonucleotides of length 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, or 120 with further single-nucleotide additions up to the longest sequence disclosed. In some embodiments, longer nucleic acids of up to 240 bases which comprise the sequences disclosed herein may be used. Moreover, the oligonucleotides of the invention do not require a symmetrical extension on either side of the central DNA domain. Similarly, the oligonucleotides of the invention as disclosed in the various tables for alteration of particular plant genes contain phosphorothioate linkages, 2′-O-methyl analog or LNA (including LNA derivatives and analogs) or any combination of these modifications just as the assay oligonucleotides do.

[0022] The present invention, however, is not limited to oligonucleotides that contain any particular nuclease resistant modification. Oligonucleotides of the invention may be altered with any combination of additional LNAs (including LNA derivatives and analogs), phosphorothioate linkages or 2′-O-methyl analogs to maximize conversion efficiency. For oligonucleotides of the invention that are longer than about 17 to about 25 bases in length, internal as well as terminal region segments of the backbone may be altered. Alternatively, simple fold-back structures at each end of a oligonucleotide or appended end groups may be used in addition to a modified backbone for conferring additional nuclease resistance.

[0023] The different oligonucleotides of the present invention preferably contain more than one of the aforementioned backbone modifications at each end. In some embodiments, the backbone modifications are adjacent to one another. However, the optimal number and placement of backbone modifications for any individual oligonucleotide will vary with the length of the oligonucleotide and the particular type of backbone modification(s) that are used. If constructs of identical sequence having phosphorothioate linkages are compared, 2, 3, 4, 5, or 6 phosphorothioate linkages at each end are preferred. If constructs of identical sequence having 2′-O-methyl base analogs are compared, 1, 2, 3 or 4 analogs are preferred. The optimal number and type of backbone modifications for any particular oligo-nucleotide useful for altering target DNA may be determined empirically by comparing the alteration efficiency of the oligonucleotide comprising any combination of the modifications to a control molecule of comparable sequence using any of the assays described herein. The optimal position(s) for oligonucleotide modifications for a maximally efficient altering oligonucleotide can be determined by testing the various modifications as compared to control molecule of comparable sequence in one of the assays disclosed herein. In such assays, a control molecule includes, e.g., a completely 2′-O-methyl substituted molecule, a completely complementary oligonucleotide, or a chimeric RNA-DNA double hairpin.

[0024] Increasing the number of phosphorothioate linkages, LNAs or 2′-O-methyl bases beyond the preferred number generally decreases the gene repair activity of a 25 nucleotide long oligonucleotide. Based on analysis of the concentration of oligonucleotide present in the extract after different time periods of incubation, it is believed that the terminal modifications impart nuclease resistance to the oligo-nucleotide thereby allowing it to survive within the cellular environment. However, this may not be the only possible mechanism by which such modifications confer greater efficiency of conversion. For example, as disclosed herein, certain modifications to oligonucleotides confer a greater improvement to the efficiency of conversion than other modifications.

[0025] Efficiency of conversion is defined herein as the percentage of recovered substrate molecules that have undergone a conversion event. Depending on the nature of the target genetic material, e.g. the genome of a cell, efficiency could be represented as the proportion of cells or clones containing an extrachromosomal element that exhibit a particular phenotype. Alternatively, representative samples of the target genetic material can be sequenced to determine the percentage that have acquired the desire change. The oligonucleotides of the invention in different embodiments can alter DNA two, three, four, five, six, seven, eight, nine, ten, twelve, fifteen, twenty, thirty, and fifty or more fold more than control oligonucleotides. Such control oligonucleotides are oligonucleotides with fully phosphorothiolated linkages, oligonucleotides that are fully substituted with 2′-O-methyl analogs, a perfectly matched oligonucleotide that is fully complementary to a target sequence or a chimeric DNA-RNA double hairpin oligonucleotide such as disclosed in U.S. Pat. No. 5,565,350.

[0026] In addition, for a given oligonucleotide length, additional modifications interfere with the ability of the oligonucleotide to act in concert with the cellular recombination or repair enzyme machinery which is necessary and required to mediate a targeted substitution, addition or deletion event in DNA. For example, fully phosphorothiolated or fully 2-O-methylated molecules are inefficient in targeted gene alteration.

[0027] The oligonucleotides of the invention as optimized for the purpose of targeted alteration of genetic material, including gene knockout or repair, are different in structure from antisense oligo-nucleotides that may possess a similar mixed chemical composition backbone. The oligonucleotides of the invention differ from such antisense oligonucleotides in chemical composition, structure, sequence, and in their ability to alter genomic DNA. Significantly, antisense oligonucleotides fail to direct targeted gene alteration. The oligonucleotides of the invention may target either strand of DNA and can include any component of the genome including, for example, intron and exon sequences. The preferred embodiment of the invention is a modified oligonucleotide that binds to the non-transcribed strand of a genomic DNA duplex. In other words, the preferred oligonucleotides of the invention target the sense strand of the DNA, i.e. the oligonucleotides of the invention are complementary to the non-transcribed strand of the target duplex DNA. The sequence of the non-transcribed strand of a DNA duplex is found in the mRNA produced from that duplex, given that mRNA uses uracil-containing nucleotides in place of thymine-containing nucleotides.

[0028] Moreover, the initial observation that single-stranded oligonucleotides comprising these modifications and lacking any particular triplex forming domain have reproducibly enhanced gene alteration activity in a variety of assay systems as compared to a chimeric RNA-DNA double-stranded hairpin control or single-stranded oligonucleotides comprising other backbone modifications was surprising. The single-stranded molecules of the invention totally lack the complementary RNA binding structure that stabilizes a normal chimeric double-stranded hairpin of the type disclosed in U.S. Pat. No. 5,565,350 yet is more effective in producing targeted base conversion as compared to such a chimeric RNA-DNA double-stranded hairpin. In addition, the molecules of the invention lack any particular triplex forming domain involved in Hoogsteen interactions with the DNA double helix and required by other known oligonucleotides in other oligonucleotide-dependant gene conversion systems. Although the lack of these functional domains was expected to decrease the efficiency of an alteration in a sequence, just the opposite occurs: the efficiency of sequence alteration using the modified oligonucleotides of the invention is higher than the efficiency of sequence alteration using a chimeric RNA-DNA hairpin targeting the same sequence alteration. Moreover, the efficiency of sequence alteration or gene conversion directed by an unmodified oligonucleotide is many times lower as compared to a control chimeric RNA-DNA molecule or the modified oligonucleotides of the invention targeting the same sequence alteration. Similarly, molecules containing at least 3 2′-O-methyl base analogs are about four to five fold less efficient as compared to an oligonucleotide having the same number of phosphorothioate linkages.

[0029] The oligonucleotides of the present invention for alteration of a single base are about 17 to about 121 nucleotides in length, preferably about 17 to about 74 nucleotides in length. Most preferably, however, the oligonucleotides of the present invention are at least about 25 bases in length, unless there are self-dimerization structures within the oligonucleotide. If the oligonucleotide has such an unfavorable structure, lengths longer than 35 bases are preferred. Oligonucleotides with modified ends both shorter and longer than certain of the exemplified, modified oligonucleotides herein function as gene repair or gene knockout agents and are within the scope of the present invention.

[0030] Once an oligomer is chosen, it can be tested for its tendency to self-dimerize, since self-dimerization may result in reduced efficiency of alteration of genetic information. Checking for self-dimerization tendency can be accomplished manually or, preferably, using a software program. One such program is Oligo Analyzer 2.0, available through Integrated DNA Technologies (Coralville, Iowa 52241) (http://www.idtdna.com); this program is available for use on the world wide web at http://www.idtdna.com/program/oligoanalyzer/oligoanalyzer.asp.

[0031] For each oligonucleotide sequence input into the program, Oligo Analyzer 2.0 reports possible self-dimerized duplex forms, which are usually only partially duplexed, along with the free energy change associated with such self-dimerization. Delta G-values that are negative and large in magnitude, indicating strong self-dimerization potential, are automatically flagged by the software as “bad”. Another software program that analyzes oligomers for pair dimer formation is Primer Select from DNASTAR, Inc., 1228 S. Park St., Madison, Wis. 53715, Phone: (608) 258-7420 (http://www.dnastar.com/products/PrimerSelect.html).

[0032] If the sequence is subject to significant self-dimerization, the addition of further sequence flanking the “repair” nucleotide can improve gene correction frequency.

[0033] Generally, the oligonucleotides of the present invention are identical in sequence to one strand of the target DNA, which can be either strand of the target DNA, with the exception of one or more targeted bases positioned within the DNA domain of the oligonucleotide, and preferably toward the middle between the modified terminal regions. Preferably, the difference in sequence of the oligonucleotide as compared to the targeted genomic DNA is located at about the middle of the oligo-nucleotide sequence. In a preferred embodiment, the oligonucleotides of the invention are complementary to the non-transcribed strand of a duplex. In other words, the preferred oligonucleotides target the sense strand of the DNA, i.e. the oligonucleotides of the invention are preferably complementary to the strand of the target DNA the sequence of which is found in the mRNA.

[0034] The oligonucleotides of the invention can include more than a single base change. In an oligonucleotide that is about a 70-mer, with at least one modified residue incorporated on the ends, as disclosed herein, multiple bases can be simultaneously targeted for change. The target bases may be up to 27 nucleotides apart and may not be changed together in all resultant plasmids in all cases. There is a frequency distribution such that the closer the target bases are to each other in the central DNA domain within the oligonucleotides of the invention, the higher the frequency of change in a given cell. Target bases only two nucleotides apart are changed together in every case that has been analyzed. The farther apart the two target bases are, the less frequent the simultaneous change. Thus, oligonucleotides of the invention may be used to repair or alter multiple bases rather than just one single base. For example, in a 74-mer oligonucleotide having a central base targeted for change, a base change event up to about 27 nucleotides away can also be effected. The positions of the altering bases within the oligonucleotide can be optimized using any one of the assays described herein. Preferably, the altering bases are at least about 8 nucleotides from one end of the oligonucleotide.

[0035] The oligonucleotides of the present invention can be introduced into cells by any suitable means. According to certain preferred embodiments, the modified oligonucleotides may be used alone. Suitable means, however, include the use of polycations, cationic lipids, liposomes, polyethylenimine (PEI), electroporation, biolistics, microinjection and other methods known in the art to facilitate cellular uptake. For plant cells, biolistic or particle bombardment methods are typically used. According to certain preferred embodiments of the present invention, isolated plant cells are treated in culture according to the methods of the invention, to mutate or repair a target gene. Alternatively, plant target DNA may be modified in vitro or in another cell type, including for example, yeast or bacterial cells and then introduced into a plant cell as, for example, a T-DNA. Plant cells thus modified may be used to regenerate the whole organism as, for example, in a plant having a desired targeted genomic change. In other instances, targeted genomic alteration, including repair or mutagenesis, may take place in vivo following direct administration of the modified, single-stranded oligonucleotides of the invention to a subject.

[0036] The single-stranded, modified oligonucleotides of the present invention have numerous applications as gene repair, gene modification, or gene knockout agents. Such oligonucleotides may be advantageously used, for example, to introduce or correct multiple point mutations. Each mutation leads to the addition, deletion or substitution of at least one base pair. The methods of the present invention offer distinct advantages over other methods of altering the genetic makeup of an organism, in that only the individually targeted bases are altered. No additional foreign DNA sequences are added to the genetic complement of the organism. Such agents may, for example, be used to develop plants with improved traits by rationally changing the sequence of selected genes in isolated cells and using these modified cells to regenerate whole plants having the altered gene. See, e.g., U.S. Pat. No. 6,046,380 and U.S. Pat. No. 5,905,185 incorporated herein by reference. Such plants produced using the compositions of the invention lack additional undesirable selectable markers or other foreign DNA sequences. Targeted base pair substitution or frameshift mutations introduced by an oligonucleotide in the presence of a cell-free extract also provides a way to modify the sequence of extrachromosomal elements, including, for example, plasmids, cosmids and artificial chromosomes. The oligonucleotides of the invention also simplify the production of plants having particular modified or inactivated genes. Altered plant model systems such as those produced using the methods and oligonucleotides of the invention are invaluable in determining the function of a gene and in evaluating drugs. The oligonucleotides and methods of the present invention may also be used to introduce molecular markers, including, for example, SNPs, RFLPs, AFLPs and CAPs.

[0037] The purified oligonucleotide compositions may be formulated in accordance with routine procedures depending on the target. For example, purified oligonucleotide can be used directly in a standard reaction mixture to introduce alterations into targeted DNA in vitro or where cells are the target as a composition adapted for bathing cells in culture or for microinjection into cells in culture. The purified oligonucleotide compositions may also be provided on coated microbeads for biolistic delivery into plant cells. Where necessary, the composition may also include a solubilizing agent. Generally, the ingredients will be supplied either separately or mixed together in single-use form, for example, as a dry, lyophilized powder or water-free concentrate. In general, dosage required for efficient targeted gene alteration will range from about 0.001 to 50,000 μg/kg target tissue, preferably between 1 to 250 μg/kg, and most preferably at a concentration of between 30 and 60 micromolar.

[0038] For cell administration, direct injection into the nucleus, biolistic bombardment, electroporation, liposome transfer and calcium phosphate precipitation may be used. In yeast, lithium acetate or spheroplast transformation may also be used. In a preferred method, the administration is performed with a liposomal transfer compound, e.g., DOTAP (Boehringer-Mannheim) or an equivalent such as lipofectin. The amount of the oligonucleotide used is about 500 nanograms in 3 micrograms of DOTAP per 100,000 cells. For electroporation, between 20 and 2000 nanograms of oligonucleotide per million cells to be electroporated is an appropriate range of dosages which can be increased to improve efficiency of genetic alteration upon review of the appropriate sequence according to the methods described herein. For biolistic delivery, microbeads are generally coated with resuspended oligonucleotides, which range of oligonucleotide to microbead concentration can be similarly adjusted to improve efficiency as determined using one of the assay methods described herein, starting with about 0.05 to 1 microgram of oligonucleotide to 25 microgram of 1.0 micrometer gold beads or similar microcarrier.

[0039] Another aspect of the invention is a kit comprising at least one oligonucleotide of the invention. The kit may comprise an additional reagent or article of manufacture. The additional reagent or article of manufacture may comprise a delivery mechanism, cell extract, a cell, or a plasmid, such as one of those disclosed in the Figures herein, for use in an assay of the invention. Alternatively, the invention includes a kit comprising an isogenic set of cells in which each cell in the kit comprises a different altered amino acid for a target protein encoded by a targeted altered gene within the cell produced according to the methods of the invention.

BRIEF DESCRIPTION OF THE DRAWINGS

[0040]
FIG. 1. Flow diagram for the generation of modified single-stranded oligonucleotides. The upper strands of chimeric oligonucleotides I and II are separated into pathways resulting in the generation of single-stranded oligonucleotides that contain (A) 2′-O-methyl RNA nucleotides or (B) phosphorothioate linkages. Fold changes in repair activity for correction of kans in the HUH7 cell-free extract are presented in parenthesis. HUH7 cells are described in Nakabayashi et al., Cancer Research 42: 3858-3863 (1982). Each single-stranded oligonucleotide is 25 bases in length and contains a G residue mismatched to the complementary sequence of the kans gene. The numbers 3, 6, 8, 10, 12 and 12.5 respectively indicate how many phosphorothioate linkages (S) or 2′-O-methyl RNA nucleotides (R) are at each end of the molecule. Hence oligo 12S/25G contains an all phosphorothioate backbone, displayed as a dotted line. Smooth lines indicate DNA residues, wavy lines indicate 2′-O-methyl RNA residues and the carat indicates the mismatched base site (G). FIG. 1(C) provides a schematic plasmid indicating the sequence of the kan chimeric double-stranded hairpin oligonucleotide (left; SEQ ID NO: 2673) and the sequence the tet chimeric double-stranded hairpin oligonucleotide used in other experiments (right; SEQ ID NO: 2674). FIG. 1(D) provides a flow chart of a kan experiment in which a chimeric double-stranded hairpin oligonucleotide (SEQ ID NO: 2673) is used. In FIG. 1(D), the Kan mutant sequence corresponds to SEQ ID NO: 2675 and SEQ ID NO: 2676; the Kan converted sequence corresponds to SEQ ID NO: 2677 and SEQ ID NO: 2678; the mutant sequence in the sequence trace corresponds to SEQ ID NO: 2679 and the converted sequences in the sequence trace correspond to SEQ ID NO: 2680.

[0041]
FIG. 2. Genetic readout system for correction of a point mutation in plasmid pKsm4021. A mutant kanamycin gene harbored in plasmid pKsm4021 is the target for correction by oligonucleotides. The mutant G is converted to a C by the action of the oligo. Corrected plasmids confer resistance to kanamycin in E.coli (DH10B) after electroporation leading to the genetic readout and colony counts. The wild type sequence corresponds to SEQ ID NO: 2681.

[0042]
FIG. 3: Target plasmid and sequence correction of a frameshift mutation by chimeric and single-stranded oligonucleotides. (A) Plasmid pTsΔ208 contains a single base deletion mutation at position 208 rendering it unable to confer tet resistance. The target sequence presented below indicates the insertion of a T directed by the oligonucleotides to re-establish the resistant phenotype. (B) DNA sequence confirming base insertion directed by Tet 3S/25G; the yellow highlight indicates the position of frameshift repair. The wild type sequence corresponds to SEQ ID NO: 2682, the mutant sequence corresponds to SEQ ID NO: 2683 and the converted sequence corresponds to SEQ ID NO: 2684. The control sequence in the sequence trace corresponds to SEQ ID NO: 2685 and the 3S/25A sequence in the sequence trace corresponds to SEQ ID NO: 2686.

[0043]
FIG. 4. DNA sequences of representative kanr colonies. Confirmation of sequence alteration directed by the indicated molecule is presented along with a table outlining codon distribution. Note that 10S/25G and 12S/25G elicit both mixed and unfaithful gene repair. The number of clones sequenced is listed in parentheses next to the designation for the single-stranded oligonucleotide. A plus (+) symbol indicates the codon identified while a figure after the (+) symbol indicates the number of colonies with a particular sequence. TAC/TAG indicates a mixed peak. Representative DNA sequences are presented below the table with yellow highlighting altered residues. The sequences in the sequence traces have been assigned numbers as follows: 3S/25G, 6S/25G and 8S/25G correspond to SEQ ID NO: 2687, 10S/25G corresponds to SEQ ID NO: 2688, 25S/25G on the lower left corresponds to SEQ ID NO: 2689 and 25S/25G on the lower right corresponds to SEQ ID NO: 2690.

[0044]
FIG. 5. Gene correction in HeLa cells. Representative oligonucleotides of the invention are co-transfected with the pCMVneo(−)FIAsH plasmid (shown in FIG. 9) into HeLa cells. Ligand is diffused into cells after co-transfection of plasmid and oligonucleotides. Green fluorescence indicates gene correction of the mutation in the antibiotic resistance gene. Correction of the mutation results in the expression of a fusion protein that carries a marker ligand binding site and when the fusion protein binds the ligand, a green fluorescence is emitted. The ligand is produced by Aurora Biosciences and can readily diffuse into cells enabling a measurement of corrected protein function; the protein must bind the ligand directly to induce fluorescence. Hence cells bearing the corrected plasmid gene appear green while “uncorrected” cells remain colorless.

[0045]
FIG. 6. Z-series imaging of corrected cells. Serial cross-sections of the HeLa cell represented in FIG. 5 are produced by Zeiss 510 LSM confocal microscope revealing that the fusion protein is contained within the cell.

[0046]
FIG. 7. Hygromycin-eGFP target plasmids. (A) Plasmid pAURHYG(ins)GFP contains a single base insertion mutation between nucleotides 136 and 137, at codon 46, of the Hygromycin B coding sequence (cds) which is transcribed from the constitutive ADH1 promoter. The target sequence presented below indicates the deletion of an A and the substitution of a C for a T directed by the oligonucleotides to re-establish the resistant phenotype. In FIG. 7A, the sequence of the normal allele corresponds to SEQ ID NO: 2691, the sequence of the targe/existing mutation corresponds to SEQ ID NO: 2692 and the sequence of the desired alteration corresponds to SEQ ID NO: 2693. (B) Plasmid pAURHYG(rep)GFP contains a base substitution mutation introducing a G at nucleotide 137, at codon 46, of the Hygromycin B coding sequence (cds). The target sequence presented below the diagram indicates the amino acid conservative replacement of G with C, restoring gene function. In FIG. 7B, the sequence of the normal allele correspond to SEQ ID NO: 2691, the sequence of the targe/existing mutation corresponds to SEQ ID NO: 2694 and the sequence of the desired alteration corresponds to SEQ ID NO: 2693.

[0047]
FIG. 8. Oligonucleotides for correction of hygromycin resistance gene. The sequence of the oligonucleotides used in experiments to assay correction of a hygromycin resistance gene are shown. DNA residues are shown in capital letters, RNA residues are shown in lowercase and nucleotides with a phosphorothioate backbone are capitalized and underlined. In FIG. 8, the sequence of HygE3T/25 corresponds to SEQ ID NO: 2695, the sequence of HygE3T/74 corresponds to SEQ ID NO: 2696, the sequence of HygE3T/74a corresponds to SEQ ID NO: 2697, the sequence of HygGG/Rev corresponds to SEQ ID NO: 2698 and the sequence of Kan70T corresponds to SEQ ID NO: 2699.

[0048]
FIG. 9. pAURNeo(−)FIAsH plasmid. This figure describes the plasmid structure, target sequence, oligonucleotides, and the basis for detection of the gene alteration event by fluorescence. In FIG. 9, the sequence of the Neo/kan target mutant corresponds to SEQ ID NO: 2675 and SEQ ID NO: 2676, the converted sequence corresponds to SEQ ID NO: 2677 and SEQ ID NO: 2678 and the FIAsH peptide sequence corresponds to SEQ ID NO: 2700.

[0049]
FIG. 10. pYESHyg(x)eGFP plasmid. This plasmid is a construct similar to the pAURHyg(x)eGFP construct shown in FIG. 7, except the promoter is the inducible GAL1 promoter. This promoter is inducible with galactose, leaky in the presence of raffinose, and repressed in the presence of dextrose.

[0050]
FIG. 11. pBI-HygeGFP plasmid. This plasmid is a construct based on the plasmids pBI101, pBI 101.2, pBI101.3 or pBI 121 available from Clontech in which HygeGFP replaces the beta-glucuronidase gene of the Clontech plasmids. The different Clontech plasmids vary by a reading frame shift relative to the polylinker, or the presence of the Cauliflower mosaic virus promoter.

[0051] The following examples are provided by way of illustration only, and are not intended to limit the scope of the invention disclosed herein.

EXAMPLE 1

Assay Method for Base Alteration and Preferred Oligonucleotide Selection

[0052] In this example, single-stranded and double-hairpin oligonucleotides with chimeric backbones (see FIG. 1 for structures (A and B) and sequences (C and D) of assay oligonucleotides) are used to correct a point mutation in the kanamycin gene of pKsm4021 (FIG. 2) or the tetracycline gene of pTsΔ208 (FIG. 3). All kan oligonucleotides share the same 25 base sequence surrounding the target base identified for change, just as all tet oligonucleotides do. The sequence is given in FIG. 1C and FIG. 1D. Each plasmid contains a functional ampicillin gene. Kanamycin gene function is restored when a G at position 4021 is converted to a C (via a substitution mutation); tetracycline gene function is restored when a deletion at position 208 is replaced by a C (via frameshift mutation). A separate plasmid, pAURNeo(−)FIAsH (FIG. 9), bearing the kans gene is used in the cell culture experiments. This plasmid was constructed by inserting a synthetic expression cassette containing a neomycin phosphotransferasea (kanamycin resistance) gene and an extended reading frame that encodes a receptor for the FIAsH ligand into the pAUR123 shuttle vector (Panvera Corp., Madison, Wis.). The resulting construct replicates in S. cerevisiae at low copy number, confers resistance to aureobasidinA and constitutively expresses either the Neo+/FIAsH fusion product (after alteration) or the truncated Neo−/FIAsH product (before alteration) from the ADH1 promoter. By extending the reading frame of this gene to code for a unique peptide sequence capable of binding a small ligand to form a fluorescent complex, restoration of expression by correction of the stop codon can be detected in real time using confocal microscopy.

[0053] Additional constructs can be made to test additional gene alteration events or for specific use in different expression systems. For example, alternative comparable plant plasmids or integration vectors such as, e.g. those based on T-DNA, can be constructed for stable expression in plant cells according to the disclosures herein. Such constructs would use a plant specific promoter such as, e.g., cauliflower mosaic virus 35S promoter, to replace the promoters directing expression of the neo, hyg or aureobasidinA resistance gene disclosed herein, including for example, in FIGS. 7B, 9 and 10 herein. Moreover, the green fluorescent protein (GFP) sequence used herein may be modified to increase expression in plant cells such as Arabidopsis and the other plants disclosed herein as described in Haseloff et al., Proc. Natl.Acad. Sci. 94(6): 2122-7 (1997), Rouwendal et al. Plant Mol. Biol. 33(6): 989-99 (1997) and Hu et al. FEBS Lett. 369(2-3): 331-4 (1995). Codon usage for optimal expression of GFP in plants results from increasing the frequency of codons with a C or a G in the third position from 32 to about 60%. Specific constructs are disclosed and can be used as follows with such plant specific alterations.

[0054] We also construct three mammalian expression vectors, pHyg(rep)eGFP, pHyg(Δ)eGFP, pHyg(ins)eGFP, that contain a substitution mutation at nucleotide 137 of the hygromycin-B coding sequence. (rep) indicates a T1374→G replacement, (Δ) represents a deletion of the G137 and (ins) represents an A insertion between nucleotides 136 and 137. All point mutations create a nonsense termination codon at residue 46. We use pHYGeGFP plasmid (Invitrogen, CA) DNA as a template to introduce the mutations into the hygromycin-eGFP fusion gene by a two step site-directed mutagenesis PCR protocol. First, we generate overlapping 5′ and a 3′ amplicons surrounding the mutation site by PCR for each of the point mutation sites. A 215 bp 5′ amplicon for the (rep), (Δ) or (ins) was generated by polymerization from oligonucleotide primer HygEGFPf (5′-AATACGACTCACTATAGG-3′; SEQ ID NO: 2701) to primer Hygrepr (5′GACCTATCCACGCCCTCC-3′; SEQ ID NO: 2702), HygΔr (5′-GACTATCCACGCCCTCC-3′; SEQ ID NO: 2703), or Hyginsr (5′-GACATTATCCACGCCCTCC-3′; SEQ ID NO: 2704), respectively. We generate a 300 bp 3′ amplicon for the (rep), (Δ) or (ins) by polymerization from oligonucleotide primers Hygrepf (5′-CTGGGATAGGTCCTGCGG-3′; SEQ ID NO: 2705), HygΔf (5′-CGTGGATAGTCCTGCGG-3′; SEQ ID NO: 2706), Hyginsf (5′-CGTGGATAATGTCCTGCGG-3′; SEQ ID NO: 2707), respectively to primer HygEGFPr (5′-AAATCACGCCATGTAGTG-3′; SEQ ID NO: 2708). We mix 20 ng of each of the resultant 5′ and 3′ overlapping amplicon mutation sets and use the mixture as a template to amplify a 523 bp fragment of the Hygromycin gene spanning the KpnI and RsrII restriction endonuclease sites. We use the Expand PCR system (Roche) to generate all amplicons with 25 cycles of denaturing at 94° C. for 10 seconds, annealing at 55° C. for 20 seconds and elongation at 68° C. for 1 minute. We digest 10 μg of vector pHYGeGFP and 5 μg of the resulting fragments for each mutation with KpnI and RsrII (NEB) and gel purify the fragment for enzymatic ligation. We ligate each mutated insert into pHYGeGFP vector at 3:1 molar ratio using T4 DNA ligase (Roche). We screen clones by restriction digest, confirm the mutation by Sanger dideoxy chain termination sequencing and purify the plasmid using a Qiagen maxiprep kit.

[0055] Oligonucleotide synthesis and cells. Chimeric oligonucleotides and single-stranded oligonucleotides (including those with the indicated modifications) are synthesized using available phosphoramidites on controlled pore glass supports. After deprotection and detachment from the solid support, each oligonucleotide is gel-purified using, for example, procedures such as those described in Gamper et al., Biochem. 39, 5808-5816 (2000) and the concentrations determined spectrophotometrically (33 or 40 μg/ml per A260 unit of single-stranded or hairpin oligomer). HUH7 cells are grown in DMEM, 10% FBS, 2 mM glutamine, 0.5% pen/strep. The E.coli strain, DH10B, is obtained from Life Technologies (Gaithersburg, Md.); DH10B cells contain a mutation in the RECA gene (recA).

[0056] Cell-free extracts. Although this portion of this example is directed to mammalian systems, similar extracts from plants can be prepared as disclosed elsewhere in this application and used as disclosed in this example. We prepare cell-free extracts from HUH7 cells or other mammalian cells, as follows. We employ this protocol with essentially any mammalian cell including, for example, H1299 cells (human epithelial carcinoma, non-small cell lung cancer), C127I (immortal murine mammary epithelial cells), MEF (mouse embryonic fibroblasts), HEC-1-A (human uterine carcinoma), HCT15 (human colon cancer), HCT116 (human colon carcinoma), LoVo (human colon adenocarcinoma), and HeLa (human cervical carcinoma). We harvest approximately 2×108 cells. We then wash the cells immediately in cold hypotonic buffer (20 mM HEPES, pH7.5; 5 mM KCl; 1.5 mM MgCl2; 1 mM DTT) with 250 mM sucrose. We then resuspend the cells in cold hypotonic buffer without sucrose and after 15 minutes we lyse the cells with 25 strokes of a Dounce homogenizer using a tight fitting pestle. We incubate the lysed cells for 60 minutes on ice and centrifuge the sample for 15 minutes at 12000×g. The cytoplasmic fraction is enriched with nuclear proteins due to the extended co-incubation of the fractions following cell breakage. We then immediately aliquote and freeze the supernatant at −80° C. We determine the protein concentration in the extract by the Bradford assay.

[0057] We also perform these experiments with cell-free extracts obtained from fungal cells, including, for example, S. cerevisiae (yeast), Ustilago maydis, and Candida albicans. For example, we grow yeast cells into log phase in 2L YPD medium for 3 days at 30° C. We then centrifuge the cultures at 5000×g, resuspend the pellets in a 10% sucrose, 50 mM Tris, 1 mM EDTA lysis solution and freeze them on dry ice. After thawing, we add KCl, spermidine and lyticase to final concentrations of 0.25 mM, 5 mM and 0.1 mg/ml, respectively. We incubate the suspension on ice for 60 minutes, add PMSF and Triton X100 to final concentrations of 0.1 mM and 0.1% and continue to incubate on ice for 20 minutes. We centrifuge the lysate at 3000×g for 10 minutes to remove larger debris. We then remove the supernatant and clarify it by centrifuging at 30000×g for 15 minutes. We then add glycerol to the clarified extract to a concentration of 10% (v/v) and freeze aliquots at −80° C. We determine the protein concentration of the extract by the Bradford assay.

[0058] Reaction mixtures of 50 μl are used, consisting of 10-30 μg protein of cell-free extract, which can be optionally substituted with purified proteins or enriched fractions, about 1.5 μg chimeric double-hairpin oligonucleotide or 0.55 μg single-stranded molecule (3S/25G or 6S/25G, see FIG. 1), and 1 μg of plasmid DNA (see FIGS. 2 and 3) in a reaction buffer of 20 mM Tris, pH 7.4, 15 mM MgCl2, 0.4 mM DTT, and 1.0 mM ATP. Reactions are initiated with extract and incubated at 30° C. for 45 min. The reaction is stopped by placing the tubes on ice and then immediately deproteinized by two phenol/chloroform (1:1) extractions. Samples are then ethanol precipitated. The nucleic acid is pelleted at 15,000 r.p.m. at 4° C. for 30 min., is washed with 70% ethanol, resuspended in 50 μl H2O, and is stored at −20° C. 5 μl of plasmid from the resuspension (˜100 ng) was transfected in 20 μl of DH10B cells by electroporation (400 V, 300 μF, 4 kΩ) in a Cell-Porator apparatus (Life Technologies). After electroporation, cells are transferred to a 14 ml Falcon snap-cap tube with 2 ml SOC and shaken at 37° C. for 1 h. Enhancement of final kan colony counts is achieved by then adding 3 ml SOC with 10 μg/ml kanamycin and the cell suspension is shaken for a further 2 h at 37° C. Cells are then spun down at 3750×g and the pellet is resuspended in 500 μl SOC. 200 μl is added undiluted to each of two kanamycin (50 μg/ml) agar plates and 200 μl of a 105 dilution is added to an ampicillin (100 μg/ml) plate. After overnight 37° C. incubation, bacterial colonies are counted using an Accucount 1000 (Biologics). Gene conversion effectiveness is measured as the ratio of the average of the kan colonies on both plates per amp colonies multiplied by 10−5 to correct for the amp dilution.

[0059] The following procedure can also be used. 5 μl of resuspended reaction mixtures (total volume 50 μl) are used to transform 20 μl aliquots of electro-competent DH10B bacteria using a Cell-Porator apparatus (Life Technologies). The mixtures are allowed to recover in 1 ml SOC at 37° C. for 1 hour at which time 50 μg/ml kanamycin or 12 μg/ml tetracycline is added for an additional 3 hours. Prior to plating, the bacteria are pelleted and resuspended in 200 μl of SOC. 100 μl aliquots are plated onto kan or tet agar plates and 100 μl of a 1031 4 dilution of the cultures are concurrently plated on agar plates containing 100 μg/ml of ampicillin. Plating is performed in triplicate using sterile Pyrex beads. Colony counts are determined by an Accu-count 1000 plate reader (Biologics). Each plate contains 200-500 ampicillin resistant colonies or 0-500 tetracycline or kanamycin resistant colonies. Resistant colonies are selected for plasmid extraction and DNA sequencing using an ABI Prism kit on an ABI 310 capillary sequencer (PE Biosystems).

[0060] Chimeric single-stranded oligonucleotides. In FIG. 1 the upper strands of chimeric oligonucleotides I and II are separated into pathways resulting in the generation of single-stranded oligo-nucleotides that contain (FIG. 1A) 2′-O-methyl RNA nucleotides or (FIG. 1B) phosphorothioate linkages. Fold changes in repair activity for correction of kans in the HUH7 cell-free extract are presented in parenthesis. Each single-stranded oligonucleotide is 25 bases in length and contains a G residue mismatched to the complementary sequence of the kans gene.

[0061] Molecules bearing 3, 6, 8, 10 and 12 phosphorothioate linkages in the terminal regions at each end of a backbone with a total of 24 linkages (25 bases) are tested in the kans system. Alternatively, molecules bearing 2, 4, 5, 7, 9 and 11 in the terminal regions at each end are tested. The results of one such experiment, presented in Table 1 and FIG. 1B, illustrate an enhancement of correction activity directed by some of these modified structures. In this illustrative example, the most efficient molecules contained 3 or 6 phosphorothioate linkages at each end of the 25-mer; the activities are approximately equal (molecules IX and X with results of 3.09 and 3.7 respectively). A reduction in alteration activity may be observed as the number of modified linkages in the molecule is further increased. Interestingly, a single-strand molecule containing 24 phosphorothioate linkages is minimally active suggesting that this backbone modification when used throughout the molecule supports only a low level of targeted gene repair or alteration. Such a non-altering, completely modified molecule can provide a baseline control for determining efficiency of correction for a specific oligonucleotide molecule of known sequence in defining the optimum oligonucleotide for a particular alteration event.

[0062] The efficiency of gene repair directed by phosphorothioate-modified, single-stranded molecules, in a length dependent fashion, led us to examine the length of the RNA modification used in the original chimera as it relates to correction. Construct III represents the “RNA-containing” strand of chimera I and, as shown in Table 1 and FIG. 2A, it promotes inefficient gene repair. But, as shown in the same figure, reducing the RNA residues on each end from 10 to 3 increases the frequency of repair. At equal levels of modification, however, 25-mers with 2′-O-methyl ribonucleotides were less effective gene repair agents than the same oligomers with phosphorothioate linkages. These results reinforce the fact that an RNA containing oligonucleotide is not as effective in promoting gene repair or alteration as a modified DNA oligonucleotide.

[0063] Repair of the kanamycin mutation requires a G→C exchange. To confirm that the specific desired correction alteration was obtained, colonies selected at random from multiple experiments are processed and the isolated plasmid DNA is sequenced. As seen in FIG. 4, colonies generated through the action of the single-stranded molecules 3S/25G (IX), 6S/25G (X) and 8S/25G (XI) respectively contained plasmid molecules harboring the targeted base correction. While a few colonies appeared on plates derived from reaction mixtures containing 25-mers with 10 or 12 thioate linkages on both ends, the sequences of the plasmid molecules from these colonies contain nonspecific base changes. In these illustrative examples, the second base of the codon is changed (see FIG. 3). These results show that modified single-strands can direct gene repair, but that efficiency and specificity are reduced when the 25-mers contain 10 or more phosphorothioate linkages at each end.

[0064] In FIG. 1, the numbers 3, 6, 8, 10, 12 and 12.5 respectively indicate how many phosphorothioate linkages (S) or 2′-O-methyl RNA nucleotides (R) are at each end of the examplified molecule although other molecules with 2, 4, 5, 7, 9 and 11 modifications at each end can also be tested. Hence oligo 12S/25G represents a 25-mer oligonucleotide which contains 12 phosphorothioate linkages on each side of the central G target mismatch base producing a fully phosphorothioate linked backbone, displayed as a dotted line. The dots are merely representative of a linkage in the figure and do not depict the actual number of linkages of the oligonucleotide. Smooth lines indicate DNA residues, wavy lines indicate 2′-O-methyl RNA residues and the carat indicates the mismatched base site (G).

[0065] Correction of a mutant kanamycin gene in cultured mammalian cells. Although this portion of this example is directed to cultured mammalian cells, comparable methods may be used using cultured plant cells or protoplasts of those cells from the plant species disclosed herein. The experiments are performed using different eukaryotic cells including plant and mammalian cells, including, for example, 293 cells (transformed human primary kidney cells), HeLa cells (human cervical carcinoma), and H1299 (human epithelial carcinoma, non-small cell lung cancer). HeLa cells are grown at 37° C. and 5% CO2 in a humidified incubator to a density of 2×105 cells/ml in an 8 chamber slide (Lab-Tek). After replacing the regular DMEM with Optimem, the cells are co-transfected with 10 μg of plasmid pAURNeo(−) FIAsH and 5 μg of modified single-stranded oligonucleotide (3S/25G) that is previously complexed with 10 μg lipofectamine, according to the manufacturer's directions (Life Technologies). The cells are treated with the liposome-DNA-oligo mix for 6 hrs at 37° C. Treated cells are washed with PBS and fresh DMEM is added. After a 16-18 hr recovery period, the culture is assayed for gene repair. The same oligonucleotide used in the cell-free extract experiments is used to target transfected plasmid bearing the kans gene. Correction of the point mutation in this gene eliminates a stop codon and restores full expression. This expression can be detected by adding a small non-fluorescent ligand that bound to a C-C-R-E-C-C sequence (SEQ ID NO: 2717) in the genetically modified carboxy terminus of the kan protein, to produce a highly fluorescent complex (FIAsH system, Aurora Biosciences Corporation). Following a 60 min incubation at room temperature with the ligand (FIAsH-EDT2), cells expressing full length kan product acquire an intense green fluorescence detectable by fluorescence microscopy using a fluorescein filter set. Similar experiments are performed using the HygeGFP target as described in Example 2 with a variety of mammalian cells, including, for example, COS-1 and COS-7 cells (African green monkey), and CHO-K1 cells (Chinese hamster ovary). The experiments are also performed with PG12 cells (rat pheochromocytoma) and ES cells (human embryonic stem cells).

[0066] Summary of experimental results. Tables 1, 2 and 3 respectively provide data on the efficiency of gene repair directed by single-stranded oligonucleotides. Table 1 presents data using a cell-free extract from human liver cells (HUH7) to catalyze repair of the point mutation in plasmid pkansm4021 (see FIG. 1). Table 2 illustrates that the oligomers are not dependent on MSH2 or MSH3 for optimal gene repair activity. Table 3 illustrates data from the repair of a frameshift mutation (FIG. 3) in the tet gene contained in plasmid pTetΔ208. Table 4 illustrates data from repair of the pkansm4021 point mutation catalyzed by plant cell extracts prepared from canola and musa (banana). Colony numbers are presented as kanr or tetr and fold increases (single strand versus double hairpin) are presented for kanr in Table 1.

[0067]
FIG. 5A is a confocal picture of HeLa cells expressing the corrected fusion protein from an episomal target. Gene repair is accomplished by the action of a modified single-stranded oligonucleotide containing 3 phosphorothioate linkages at each end (3S/25G). FIG. 5B represents a “Z-series” of HeLa cells bearing the corrected fusion gene. This series sections the cells from bottom to top and illustrates that the fluorescent signal is “inside the cells”.

[0068] Results. In summary, we have designed a novel class of single-stranded oligonucleotides with backbone modifications at the termini and demonstrate gene repair/conversion activity in mammalian and plant cell-free extracts. We confirm that the all DNA strand of the RNA-DNA double-stranded double hairpin chimera is the active component in the process of gene repair. In some cases, the relative frequency of repair by the novel oligonucleotides of the invention is elevated approximately 3-4-fold in certain embodiments when compared to frequencies directed by chimeric RNA-DNA double hairpin oligonucleotides.

[0069] This strategy centers around the use of extracts from various sources to correct a mutation in a plasmid using a modified single-stranded or a chimeric RNA-DNA double hairpin oligonucleotide. A mutation is placed inside the coding region of a gene conferring antibiotic resistance in bacteria, here kanamycin or tetracycline. The appearance of resistance is measured by genetic readout in E.coli grown in the presence of the specified antibiotic. The importance of this system is that both phenotypic alteration and genetic inheritance can be measured. Plasmid pKsm4021 contains a mutation (T→G) at residue 4021 rendering it unable to confer antibiotic resistance in E.coli. This point mutation is targeted for repair by oligonucleotides designed to restore kanamycin resistance. To avoid concerns of plasmid contamination skewing the colony counts, the directed correction is from G→C rather than G→T (wild-type). After isolation, the plasmid is electroporated into the DH10B strain of E.coli, which contains inactive RecA protein. The number of kanamycin colonies is counted and normalized by ascertaining the number of ampicillin colonies, a process that controls for the influence of electroporation. The number of colonies generated from three to five independent reactions was averaged and is presented for each experiment. A fold increase number is recorded to aid in comparison.

[0070] The original RNA-DNA double hairpin chimera design, e.g., as disclosed in U.S. Pat. No. 5,565,350, consists of two hybridized regions of a single-stranded oligonucleotide folded into a double hairpin configuration. The double-stranded targeting region is made up of a 5 base pair DNA/DNA segment bracketed by 10 base pair RNA/DNA segments. The central base pair is mismatched to the corresponding base pair in the target gene. When a molecule of this design is used to correct the kans mutation, gene repair is observed (I in FIG. 1A). Chimera II (FIG. 1B) differs partly from chimera I in that only the DNA strand of the double hairpin is mismatched to the target sequence. When this chimera was used to correct the kans mutation, it was twice as active. In the same study, repair function could be further increased by making the targeting region of the chimera a continuous RNA/DNA hybrid.

[0071] Frame shift mutations are repaired. By using plasmid pTsΔ208, described in FIG. 1(C) and FIG. 3, the capacity of the modified single-stranded molecules that showed activity in correcting a point mutation, can be tested for repair of a frameshift. To determine efficiency of correction of the mutation, a chimeric oligonucleotide (Tet I), which is designed to insert a T residue at position 208, is used. A modified single-stranded oligonucleotide (Tet IX) directs the insertion of a T residue at this same site. FIG. 3 illustrates the plasmid and target bases designated for change in the experiments. When all reaction components are present (extract, plasmid, oligomer), tetracycline resistant colonies appear. The colony count increases with the amount of oligonucleotide used up to a point beyond which the count falls off (Table 3). No colonies above background are observed in the absence of either extract or oligonucleotide, nor when a modified single-stranded molecule bearing perfect complementarity is used. FIG. 3 represents the sequence surrounding the target site and shows that a T residue is inserted at the correct site. We have isolated plasmids from fifteen colonies obtained in three independent experiments and each analyzed sequence revealed the same precise nucleotide insertion. These data suggest that the single-stranded molecules used initially for point mutation correction can also repair nucleotide deletions.

[0072] Comparison of phosphorothioate oligonucleotides to 2′-O-methyl substituted oligonucleotides. From a comparison of molecules VII and XI, it is apparent that gene repair is more subject to inhibition by RNA residues than by phosphorothioate linkages. Thus, even though both of these oligonucleotides contain an equal number of modifications to impart nuclease resistance, XI (with 16 phosphorothioate linkages) has good gene repair activity while VII (with 16 2′-O-methyl RNA residues) is inactive. Hence, the original chimeric double hairpin oligonucleotide enabled correction directed, in large part, by the strand containing a large region of contiguous DNA residues.

[0073] Oligonucleotides can target multiple nucleotide alterations within the same template. The ability of individual single-stranded oligonucleotides to correct multiple mutations in a single target template is tested using the plasmid pKsm4021 and the following single-stranded oligonucleotides modified with 3 phosphorothioate linkages at each end (indicated as underlined nucleotides): Oligo1 is a 25-mer with the sequence TTCGATAAGCCTATGCTGACCCGTG (SEQ ID NO: 2709) corrects the original mutation present in the kanamycin resistance gene of pKsm4021 as well as directing another alteration 2 basepairs away in the target sequence (both indicated in boldface); Oligo2 is a 70-mer with the 5′-end sequence TTCGGCTACGACTGGGCACAACAGACAATTGGC (SEQ ID NO: 2710) with the remaining nucleotides being completely complementary to the kanamycin resistance gene and also ending in 3 phosphorothioate linkages at the 3′ end. Oigo2 directs correction of the mutation in pKsm4021 as well as directing another alteration 21 basepairs away in the target sequence (both indicated in boldface).

[0074] We also use additional oligonucleotides to assay the ability of individual oligonucleotides to correct multiple mutations in the pKsM4021 plasmid. These include, for example, a second 25-mer that alters two nucleotides that are three nucleotides apart with the sequence 5′-TTGTGCCCAGTCGTATCCGAATAGC-3′ (SEQ ID NO: 2711); a 70-mer that alters two nucleotides that are 21 nucleotides apart with the sequence 5′-CATCAGAGCAGCCAATTGTCTGTTGTGCCCAGTCGTAGCCGAATAGCCTCTCCACCCAAGCGGCCGGAGA-3′ (SEQ ID NO: 2712); and another 70-mer that alters two nucleotides that are 21 nucleotides apart with the sequence 5′-GCTGACAGCCGGAACACGGCGGCATCAGAGCAGCCAATTGTCTGTTGTGCCCAGTCGTAGCCGMTAGCCT-3′ (SEQ ID NO: 2713). The nucleotides in the oligonucleotides that direct alteration of the target sequence are underlined and in boldface. These oligonucleotides are modified in the same way as the other oligonucleotides of the invention.

[0075] We assay correction of the original mutation in pKsm4021 by monitoring kanamycin resistance (the second alterations which are directed by Oligo2 and Oligo3 are silent with respect to the kanamycin resistance phenotype). In addition, in experiments with Oligo2, we also monitor cleavage of the resulting plasmids using the restriction enzyme Tsp5091 which cuts at a specific site present only when the second alteration has occurred (at ATT in Oligo2). We then sequence these clones to determine whether the additional, silent alteration has also been introduced. The results of an analysis are presented below:

2Oligo 1 (25-mer)Oligo 2 (70-mer)Clones with both sites changed97Clones with a single site changed02Clones that were not changed41

[0076] Nuclease sensitivity of unmodified DNA oligonucleotide. Electrophoretic analysis of nucleic acid recovered from the cell-free extract reactions conducted here confirm that the unmodified single-stranded 25-mer did not survive incubation whereas greater than 90% of the terminally modified oligos did survive (as judged by photo-image analyses of agarose gels).

[0077] Plant extracts direct repair. The modified single-stranded constructs can be tested in plant cell extracts. We have observed gene alteration using extracts from multiple plant sources, including, for example, Arabidopsis, tobacco, banana, maize, soybean, canola, wheat, spinach as well as spinach chloroplast extract or extracts made from other plant cells disclosed herein. We prepare the extracts by grinding plant tissue or cultured cells under liquid nitrogen with a mortar and pestle. We extract 3 ml of the ground plant tissue with 1.5 ml of extraction buffer (20 mM HEPES, pH7.5; 5 mM KCl; 1.5 mM MgCl2; 10 mM DTT; and 10% [v/v] glycerol). Some plant cell-free extracts also include about 1% (w/v) PVP. We then homogenize the samples with 15 strokes of a Dounce homogenizer. Following homogenization, we incubate the samples on ice for 1 hour and centrifuge at 3000×g for 5 minutes to remove plant cell debris. We then determine the protein concentration in the supernatants (extracts) by Bradford assay. We dispense 100 μg (protein) aliquots of the extracts which we freeze in a dry ice-ethanol bath and store at −80° C.

[0078] We describe experiments using two sources here: a dicot (canola) and a monocot (banana, Musa acuminata cv. Rasthali). Each vector directs gene repair of the kanamycin mutation (Table 4); however, the level of correction is elevated 2-3 fold relative to the frequency observed with the chimeric oligonucleotide. These results are similar to those observed in the mammalian system wherein a significant improvement in gene repair occurred when modified single-stranded molecules were used.

[0079] Tables are attached hereto.

3TABLE IGene repair activity is directed by single-stranded oligonucleotides.OligonucleotidePlasmidExtract (ug)kanr coloniesFold increaseIpKSm402110300I↓20418 1.0 ×II↓10537II↓20748 1.78 ×III↓103III↓205 0.01 ×IV↓10112IV↓2096 0.22 ×V↓10217V↓20342 0.81 ×VI↓106VI↓20390.093 ×VII↓100VII↓200 0 ×VIII↓103VIII↓205 0.01 ×IX↓10936IX↓201295 3.09 ×X↓101140X↓201588 3.7 ×XI↓10480XI↓20681 1.6 ×XII↓1018XII↓20250.059 ×XIII↓100XIII↓2040.009 ×—↓200I↓—0

[0080] Plasmid pKSm4021 (1 μg), the indicated oligonucleotide (1.5 μg chimeric oligonucleotide or 0.55 μg single-stranded oligonucleotide; molar ratio of oligo to plasmid of 360 to 1) and either 10 or 20 μg of HUH7 cell-free extract were incubated 45 min at 37° C. Isolated plasmid DNA was electroporated into E. coli (strain DH10B) and the number of kanr colonies counted. The data represent the number of kanamycin resistant colonies per 106 ampicillin resistant colonies generated from the same reaction and is the average of three experiments (standard deviation usually less than +/−15%). Fold increase is defined relative to 418 kanr colonies (second reaction) and in all reactions was calculated using the 20 μg sample.

4TABLE IIModified single-stranded oligomers are not dependent on MSH2or MSH3 for optimal gene repair activity.A.OligonucleotidePlasmidExtractkanr coloniesIX (3S/25G)↓HUH7637X (6S/25G)↓HUH7836IX↓MEF2−/−781X↓MEF2−/−676IX↓MEF3−/−582X↓MEF3−/−530IX↓MEF+/+332X↓MEF+/+497—↓MEF2−/−10—↓MEF3−/−5—↓MEF+/+14

[0081] Chimeric oligonucleotide (1.5 μg) or modified single-stranded oligonucleotide (0.55 μg) was incubated with 1 μg of plasmid pKSm4021 and 20 μg of the indicated extracts. MEF represents mouse embryonic fibroblasts with either MSH2 (2−/−) or MSH3 (3−/−) deleted. MEF+/+ indicates wild-type mouse embryonic fibroblasts. The other reaction components were then added and processed through the bacterial readout system. The data represent the number of kanamycin resistant colonies per 106 ampicillin resistant colonies.

5TABLE IIIFrameshift mutation repair is directed bysingle-stranded oligonucleotidesOligonucleotidePlasmidExtracttetr coloniesTet IX (3S/25A; 0.5 μg)pTSΔ208 (1 μg)—0—↓20 μg0Tet IX (0.5 μg)↓↓48Tet IX (1.5 μg)↓↓130Tet IX (2.0 μg)↓↓68Tet I (chimera; 1.5 μg)↓↓48

[0082] Each reaction mixture contained the indicated amounts of plasmid and oligonucleotide. The extract used for these experiments came from HUH7 cells. The data represent the number of tetracycline resistant colonies per 106 ampicillin resistant colonies generated from the same reaction and is the average of 3 independent experiments. Tet I is a chimeric oligonucleotide and Tet IX is a modified single-stranded oligonucleotide that are designed to insert a T residue at position 208 of pTsΔ208. The oligonucleotides are equivalent to structures I and IX in FIG. 2.

6TABLE IVPlant cell-free extracts support gene repair bysingle-stranded oligonucleotidesOligonucleotidePlasmidExtractkanr coloniesII (chimera)pKSm402l30 μgCanola337IX (3S/25G)↓Canola763X (6S/25G)↓Canola882II↓Musa203IX↓Musa343X↓Musa746—↓Canola0—↓Musa0IX↓—Canola0X↓—Musa0

[0083] Canola or Musa cell-free extracts were tested for gene repair activity on the kanamycin-sensitive gene as previously described in (18). Chimeric oligonucleotide II (1.5 μg) and modified single-stranded oligonucleotides IX and X (0.55 μg) were used to correct pKSm4021. Total number of kanr colonies are present per 107 ampicillin resistant colonies and represent an average of four independent experiments.

7TABLE VGene repair activity in cell-free extracts prepared from yeast(Saccharomyces cerevisiae)Cell-typePlasmidChimeric OligoSS Oligokanr/ampr × 106Wild typepKansm40211 μg0.36Wild type↓1 μg0.81ΔRAD52↓1 μg10.72ΔRAD52↓1 μg17.41ΔPMS1↓1 μg2.02ΔPMS1↓1 μg3.23In this experiment, the kans gene in pKans4021 is corrected by either a chimeric double-hairpin oligonucleotide or a single-stranded oligonucleotide containing three thioate linkages at each end (3S/25G).

EXAMPLE 2

Yeast Cell Targeting Assay Method for Base Alteration and Preferred Oligonucleotide Selection

[0084] In this example, single-stranded oligonucleotides with modified backbones and double-hairpin oligonucleotides with chimeric, RNA-DNA backbones are used to measure gene repair using two episomal targets with a fusion between a hygromycin resistance gene and eGFP as a target for gene repair. These plasmids are pAURHYG(rep)GFP, which contains a point mutation in the hygromycin resistance gene (FIG. 7), pAURHYG(ins)GFP, which contains a single-base insertion in the hygromycin resistance gene (FIG. 7) and pAURHYG(Δ)GFP which has a single base deletion. We also use the plasmid containing a wild-type copy of the hygromycin-eGFP fusion gene, designated pAURHYG(wt)GFP, as a control. These plasmids also contain an aureobasidinA resistance gene. In pAURHYG(rep)GFP, hygromycin resistance gene function and green fluorescence from the eGFP protein are restored when a G at position 137, at codon 46 of the hygromycin B coding sequence, is converted to a C thus removing a premature stop codon in the hygromycin resistance gene coding region. In pAURHYG(ins)GFP, hygromycin resistance gene function and green fluorescence from the eGFP protein are restored when an A inserted between nucleotide positions 136 and 137, at codon 46 of the hygromycin B coding sequence, is deleted and a C is substituted for the T at position 137, thus correcting a frameshift mutation and restoring the reading frame of the hygromycin-eGFP fusion gene.

[0085] We synthesize the set of three yeast expression constructs pAURHYG(rep)eGFP, pAURHYG(Δ)eGFP, pAURHYG(ins)eGFP, that contain a point mutation at nucleotide 137 of the hygromycin-B coding sequence as follows. (rep) indicates a T137→G replacement, (Δ) represents a deletion of the G137 and (ins) represents an A insertion between nucleotides 136 and 137. We construct this set of plasmids by excising the respective expression cassettes by restriction digest from pHyg(x)EGFP and ligation into pAUR123 (Panvera, Calif.). We digest 10 μg pAUR123 vector DNA, as well as, 10 μg of each pHyg(x)EGFP construct with KpnI and SaII (NEB). We gel purify each of the DNA fragments and prepare them for enzymatic ligation. We ligate each mutated insert into pHygEGFP vector at 3:1 molar ratio using T4 DNA ligase (Roche). We screen clones by restriction digest, confirm by Sanger dideoxy chain termination sequencing and purify using a Qiagen maxiprep kit.

[0086] We use this system to assay the ability of five oligonucleotides (shown in FIG. 8) to support correction under a variety of conditions. The oligonucleotides which direct correction of the mutation in pAURHYG(rep)GFP can also direct correction of the mutation in pAURHYG(ins)GFP. Three of the four oligonucleotides (HygE3T/25, HygE3T/74 and HygGG/Rev) share the same 25-base sequence surrounding the base targeted for alteration. HygGG/Rev is an RNA-DNA chimeric double hairpin oligonucleotide of the type described in the prior art. One of these oligonucleotides, HygE3T/74, is a 74-base oligonucleotide with the 25-base sequence centrally positioned. The fourth oligonucleotide, designated HygE3T/74α, is the reverse complement of HygE3T/74. The fifth oligonucleotide, designated Kan70T, is a non-specific, control oligonucleotide which is not complementary to the target sequence. Alternatively, an oligonucleotide of identical sequence but lacking a mismatch to the target or a completely thioate modified oligonucleotide or a completely 2-O-methylated modified oligonucleotide may be used as a control. Alternatively, oligonucleotides containing one, two, three, four, five, six, eight, ten or more LNA modifications on at least one of the two termini (and preferrably the 3′ terminus) may be used in different embodiments.

[0087] Oligonucleotide synthesis and cells. We synthesized and purified the chimeric, double-hairpin oligonucleotides and single-stranded oligonucleotides (including those with the indicated modifications) as described in Example 1. Plasmids used for assay were maintained stably in yeast (Saccharomyces cerevisiae) strain LSY678 MAT α at low copy number under aureobasidin selection. Plasmids and oligonucleotides are introduced into yeast cells by electroporation as follows: to prepare electrocompetent yeast cells, we inoculate 10 ml of YPD media from a single colony and grow the cultures overnight with shaking at 300 rpm at 30° C. We then add 30 ml of fresh YPD media to the overnight cultures and continue shaking at 30° C. until the OD600 was between 0.5 and 1.0 (3-5 hours). We then wash the cells by centrifuging at 4° C. at 3000 rpm for 5 minutes and twice resuspending the cells in 25 ml ice-cold distilled water. We then centrifuge at 4° C. at 3000 rpm for 5 minutes and resuspend in 1 ml ice-cold 1M sorbitol and then finally centrifuge the cells at 4° C. at 5000 rpm for 5 minutes and resuspend the cells in 120 μl 1M sorbitol. To transform electrocompetent cells with plasmids or oligonucleotides, we mix 40 μl of cells with 5 μg of nucleic acid, unless otherwise stated, and incubate on ice for 5 minutes. We then transfer the mixture to a 0.2 cm electroporation cuvette and electroporate with a BIO-RAD Gene Pulser apparatus at 1.5 kV, 25 μF, 200 Ω for one five-second pulse. We then immediately resuspend the cells in 1 ml YPD supplemented with 1M sorbitol and incubate the cultures at 30° C. with shaking at 300 rpm for 6 hours. We then spread 200 μl of this culture on selective plates containing 300 μg/ml hygromycin and spread 200 μl of a 105 dilution of this culture on selective plates containing 500 ng/ml aureobasidinA and/or and incubate at 30° C. for 3 days to allow individual yeast colonies to grow. We then count the colonies on the plates and calculate the gene conversion efficiency by determining the number of hygromycin resistance colonies per 105 aureobasidinA resistant colonies.

[0088] Frameshift mutations are repaired in yeast cells. We test the ability of the oligonucleotides shown in FIG. 8 to correct a frameshift mutation in vivo using LSY678 yeast cells containing the plasmid pAURHYG(ins)GFP. These experiments, presented in Table 6, indicate that these oligonucleotides can support gene correction in yeast cells. These data reinforce the results described in Example 1 indicating that oligonucleotides comprising phosphorothioate linkages facilitate gene correction much more efficiently than control duplex, chimeric RNA-DNA oligonucleotides. This gene correction activity is also specific as transformation of cells with the control oligonucleotide Kan70T produced no hygromycin resistant colonies above background and thus Kan70T did not support gene correction in this system. In addition, we observe that the 74-base oligonucleotide (HygE3T/74) corrects the mutation in pAURHYG(ins)GFP approximately five-fold more efficiently than the 25-base oligonucleotide (HygE3T/25). We also perform control experiments with LSY678 yeast cells containing the plasmid pAURHYG(wt)GFP. With this strain we observed that even without added oligonucleotides, there are too many hygromycin resistant colonies to count.

[0089] We also use additional oligonucleotides to assay the ability of individual oligonucleotides to correct multiple mutations in the pAURHYG(x)eGFP plasmid. These include, for example, one that alters two basepairs that are 3 nucleotides apart is a 74-mer with the sequence 5′-CTCGTGCTTTCAGCTTCGATGTAGGAGGGCGTGGGTACGTCCTGCGGGTAAATAGCTGCGCCGATGGTTTCTAC-3′ (SEQ ID NO: 2714); a 74-mer that alters two basepairs that are 15 nucleotides apart with the sequence 5′-CTCGTGCTTTCAGCTTCGATGTAGGAGGGCGTGGATACGTCCTGCGGGTAAACAGCTGCGCCGATGGTTTCTAC-3′ (SEQ ID NO: 2715); and a 74-mer that alters two basepairs that are 27 nucleotides apart with the sequence 5′-CTCGTGCTTTCAGCTTCGATGTAGGAGGGCGTGGATACGTCCTGCGGGTAAATAGCTGCGCCGACGGTTTCTAC (SEQ ID NO: 2716). The nucleotides in these oligonucleotides that direct alteration of the target sequence are underlined and in boldface. These oligonucleotides are modified in the same ways as the other oligonucleotides of the invention.

[0090] Oligonucleotides targeting the sense strand direct gene correction more efficiently. We compare the ability of single-stranded oligonucleotides to target each of the two strands of the target sequence of both pAURHYG(ins)GFP and pAURHYG(rep)GFP. These experiments, presented in Tables 7 and 8, indicate that an oligonucleotide, HygE3T/74α, with sequence complementary to the sense strand (i.e. the strand of the target sequence that is identical to the mRNA) of the target sequence facilitates gene correction approximately ten-fold more efficiently than an oligonucleotide, HygE3T/74, with sequence complementary to the non-transcribed strand which serves as the template for the synthesis of RNA. As indicated in Table 7, this effect was observed over a range of oligonucleotide concentrations from 0-3.6 μg, although we did observe some variability in the difference between the two oligonucleotides (indicated in Table 7 as a fold difference between HygE3T/74α and HygE3T/74). Furthermore, as shown in Table 8, we observe increased efficiency of correction by HygE3T/74α relative to HygE3T/74 regardless of whether the oligonucleotides were used to correct the base substitution mutation in pAURHYG(rep)GFP or the insertion mutation in pAURHYG(ins)GFP. The data presented in Table 8 further indicate that the single-stranded oligonucleotides correct a base substitution mutation more efficiently than an insertion mutation. However, this last effect was much less pronounced and the oligonucleotides of the invention are clearly able efficiently to correct both types of mutations in yeast cells. In addition, the role of transcription is investigated using plasmids with inducible promoters such as that described in FIG. 10.

[0091] Optimization of oligonucleotide concentration. To determine the optimal concentration of oligonucleotide for the purpose of gene alteration, we test the ability of increasing concentrations of Hyg3T/74α to correct the mutation in pAURHYG(rep)GFP contained in yeast LSY678. We chose this assay system because our previous experiments indicated that it supports the highest level of correction. However, this same approach could be used to determine the optimal concentration of any given oligonucleotide. We test the ability of Hyg3T/74α to correct the mutation in pAURHYG(rep)GFP contained in yeast LSY678 over a range of oligonucleotide concentrations from 0-10.0 μg. As shown in Table 9, we observe that the correction efficiency initially increases with increasing oligonucleotide concentration, but then declines at the highest concentration tested.

[0092] Tables are attached hereto.

8TABLE 6Correction of an insertion mutation in pAURHYG(ins)GFP byHygGG/Rev, HygE3T/25 and HygE3T/74Colonies onColonies onCorrectionOligonucleotide TestedHygromycinAureobasidin (/105)EfficiencyHygGG/Rev31570.02HygE3T/25641470.44HygE3T/742801741.61Kan70T0——

[0093]

9

TABLE 7

An oligonucleotide targeting the sense strand of the target sequence

corrects more efficiently.

Colonies per

hygromycin plate

Amount of Oligonucleotide (μg)
HygE3T/74
HygE3T/74α

0
0
0

0.6
24
128 (8.4x)*

1.2
69
140 (7.5x)*

2.4
62
167 (3.8x)*

3.6
29
367 (15x)*

*The numbers in parentheses represent the fold increase in efficiency for targeting the non-transcribed strand as compared to the other strand of a DNA duplex that encodes a protein.

[0094]

10

TABLE 8

Correction of a base substitution mutation is more efficient than correction

of a frame shift mutation.

Oligonucleotide
Plasmid tested (contained in LSY678)

Tested (5 μg)
pAURHYG(ins)GFP
pAURHYG(rep)GFP

HygE3T/74
72
277

HygE3T/74α
1464
2248

Kan70T
0
0

[0095]

11

TABLE 9

Optimization of oligonucleotide concentration in electroporated yeast cells.

Colonies on
Colonies on
Correction

Amount (μg)
hygromycin
aureobasidin (/105)
efficiency

0
0
67
0

1.0
5
64
0.08

2.5
47
30
1.57

5.0
199
33
6.08

7.5
383
39
9.79

10.0
191
33
5.79

EXAMPLE 3

Cultured Cell Manipulation

[0096] Although disclosure in this example is directed to use of stem cells or human blood cells and microinjection, the microinjection procedures may also be used with cultured plant cells or protoplasts using any plant species, including those disclosed herein. Mononuclear cells are isolated from human umbilical cord blood of normal donors using Ficoll Hypaque (Pharmacia Biotech, Uppsala, Sweden) density centrifugation. CD34+ cells are immunomagnetically purified from mononuclear cells using either the progenitor or Multisort Kits (Miltenyi Biotec, Auburn, Calif.). Lin−CD38− cells are purified from the mononuclear cells using negative selection with StemSep system according to the manufacturer's protocol (Stem Cell Technologies, Vancouver, Calif.). Cells used for microinjection are either freshly isolated or cryopreserved and cultured in Stem Medium (S Medium) for 2 to 5 days prior to microinjection. S Medium contains Iscoves' Modified Dulbecc's Medium without phenol red (IMDM) with 100 μg/ml glutamine/penicillin/streptomycin, 50 mg/ml bovine serum albumin, 50 μg/ml bovine pancreatic insulin, 1 mg/ml human transferrin, and IMDM; Stem Cell Technologies), 40 μg/ml low-density lipoprotein (LDL; Sigma, St. Louis, Mo.), 50 mM HEPEs buffer and 50 μM 2-mercaptoethanol, 20 ng/ml each of thrombopoietin, flt-3 ligand, stem cell factor and human IL-6 (Pepro Tech Inc., Rocky Hill, N.J.). After microinjection, cells are detached and transferred in bulk into wells of 48 well plates for culturing.

[0097] 35 mm dishes are coated overnight at 4° C. with 50 μg/ml Fibronectin (FN) fragment CH-296 (Retronectin; TaKaRa Biomedicals, Panvera, Madison, Wis.) in phosphate buffered saline and washed with IMDM containing glutamine/penicillin/streptomycin. 300 to 2000 cells are added to cloning rings and attached to the plates for 45 minutes at 37° C. prior to microinjection. After incubation, cloning rings are removed and 2 ml of S Medium are added to each dish for microinjection. Pulled injection needles with a range of 0.22 μm to 0.3 μm outer tip diameter are used. Cells are visualized with a microscope equipped with a temperature controlled stage set at 37° C. and injected using an electronically interfaced Eppendorf Micromanipulator and Transjector. Successfully injected cells are intact, alive and remain attached to the plate post injection. Molecules that are flourescently labeled allow determination of the amount of oligonucleotide delivered to the cells.

[0098] For in vitro erythropoiesis from Lin−CD38− cells, the procedure of Malik, 1998 can be used. Cells are cultured in ME Medium for 4 days and then cultured in E Medium for 3 weeks. Erythropoiesis is evident by glycophorin A expression as well as the presence of red color representing the presence of hemoglobin in the cultured cells. The injected cells are able to retain their proliferative capacity and the ability to generate myeloid and erythoid progeny. CD34+ cells can convert a normal A (βA) to sickle T (βS) mutation in the β-globin gene or can be altered using any of the oligonucleotides of the invention herein for correction or alteration of a normal gene to a mutant gene. Alternatively, stem cells can be isolated from blood of humans having genetic disease mutations and the oligonucleotides of the invention can be used to correct a defect or to modify genomes within those cells.

[0099] Alternatively, non-stem cell populations of cultured cells can be manipulated using any method known to those of skill in the art including, for example, the use of polycations, cationic lipids, liposomes, polyethylenimine (PEI), electroporation, biolistics, calcium phosphate precipitation, or any other method known in the art.

[0100] Biolistic delivery of oligonucleotide into plant cells may be accomplished according to the following method. One milliliter of packed cell volume of plant cell suspensions are subcultured onto plates containing solid medium [with Murashige and Skoog salts from Gibco/BRL, 500 mg/liter Mes, 1 mg/liter thiamin, 100 mg/liter myo-inositol, 180 mg/liter KH2PO4, 2.21 mg/liter 2,4-dichlorophenoxyacetic acid (2,4-D), and 30 g/liter sucrose (pH 5.7) and having 8 g/liter agar-agar from Sigma added before autoclaving]. By using a helium-driven particle gun such as that from BioRad and following manufacturers directions, oligonucleotides may be introduced to cells after precipitation onto 1 micrometer or comparable gold microcarriers (Bio-Rad). To precipitate onto microcarriers, 35 microliters of a particle suspension (60 mg of microcarriers per ml of 100% ethanol) is transferred to a 1.5 ml microcentrifuge tube, which is agitated on a vortex mixer. Then 40 microliter of resuspended oligonucleotide (60 ng/microliter water) is added; then 75 microliter of ice-cold 2.5 M CaCl2 is added; then 75 microliter of ice-cold 0.1 M spermidine is added. The tube is mixed vigorously or a vortex mixer for 10 min at room temperature. The particles are allowed to settle for 10 min and are centrifuged at 11,750 g for 30 sec. The supernatant is removed and the particles are resuspended in 50 microliter of 100% ethanol. An aliquot of 10 microliter of the resuspended particles are applied to each macro-projectile which is used to bombard each plate once at 900 psi (1 psi=6.89 kPa) with a gap distance (distance from power source to macroprojectile) of 1 cm and a target distance (distance from microprojectile launch site to target material) of 10 cm.

[0101] An alternative method of delivery can be used as follows. Cultured cells are suspended in liquid N6 medium and then plated on a VWR Scientific glass fiber filter. About 0.4 microgram of oligonucleotide are precipitated with 15 microliter of 2.5 mM CaCl2 and 5 microliter of 0.1 M spermidine onto 25 microgram of 1.0 micrometer gold particles. Microprojectile bombardment is performed by using a Bio-Rad PDS-1000 He particle delivery system or comparable machine following manufacturers instructions. Alterations in oligonucleotide concentrations can be employed to determine the optimum concentration of oligonucleotide according to the procedures described herein for any particular oligonucleotide of the invention.

[0102] Alternatively, the oligonucleotide of the invention may be delivered to a plant cell by electroporation of a protoplast derived from a plant part. The protoplasts may be formed by enzymatic treatment of a plant part, particularly a leaf, according to techniques such as those in Gallois et al., Methods in Molecular Biology 55: 89-107 by Humana Press. Such conditions for electroporation use about 3×105 protoplasts in a total volume of about 0.3 ml with a concentration of oligonucleotide of between 0.6 to 4 microgram per ml.

EXAMPLE 4

Plant Cells

[0103] The oligonucleotides of the invention can also be used to repair or direct a mutagenic event in plants and animal cells. Although little information is available on plant mutations amongst natural cultivars, the oligonucleotides of the invention can be used to produce “knock out” mutations by modification of specific amino acid codons to produce stop codons (e.g., a CAA codon specifying Gln can be modified at a specific site to TAA; a AAG codon specifying Lys can be modified to UAG at a specific site; and a CGA codon for Arg can be modified to a UGA codon at a specific site). Such base pair changes will terminate the reading frame and produce a defective truncated protein, shortened at the site of the stop codon.

[0104] Alternatively, frameshift additions or deletions can be directed into the genome at a specific sequence to interrupt the reading frame and produce a garbled downstream protein. Such stop or frameshift mutations can be introduced to determine the effect of knocking out the protein in either plant or animal cells.

[0105] For introduction of a T-DNA, including the T-DNA in the plasmid of FIG. 11, into a plant cell, Agrobacterium tumefaciens is used. These techniques are routine standard techniques known in the art. For example, one method follows. We transform A. tumefaciens is transformed by electroporation (using a BioRad Gene Pulser™). Competent A. tumefaciens is prepared using a method similar to that of preparing competent E. coli by suspending a freshly grown culture three times in ice-cold water and a final resuspension in 10% glycerol. Electroporation conditions are a 0.2 cm gap cuvette at a setting of 25 μF,200 Ω and2.5 kV.

[0106]

A. tumefaciens
containing a plasmid with a T-DNA is then used to introduce the T-DNA into a plant cell using routine standard techniques known in the art. For example, we transform Arabidopsis by vacuum infiltration or by dipping flowers in an Agrobacterium solution containing a surfactant, e.g. L-77. Seeds are then collected, grown and screened for presence of the T-DNA. Alternatively, Agrobacterium can be used to transform callus tissue and the callus tissue can then be used to regenerate transformed plants.

[0107] All publications and patent applications cited in this specification are herein incorporated by reference as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.

[0108] Notes on the Tables Presented Below:

[0109] Each of the following tables presents, for the specified gene, a plurality of mutations that are known to confer a relevant phenotype and, for each mutation, the oligonucleotides that can be used to correct the respective mutation site-specifically in the genome according to the present invention.

[0110] The left-most column identifies each alteration or mutation and the phenotype that the alteration/mutation confers.

[0111] For most entries, the mutation/alteration is identified at both the nucleic acid and protein level. At the amino acid level, mutations are presented according to the following standard nomenclature. The centered number identifies the position of the mutated codon in the protein sequence; to the left of the number is the wild type residue and to the right of the number is the mutant codon. Terminator codons are shown as “TERM”. At the nucleic acid level, the entire triplet of the wild type and mutated codons is shown.

[0112] The middle column presents, for each mutation, four oligonucleotides capable of repairing the mutation site-specifically in the genome or in cloned DNA including DNA in artificial chromosomes, episomes, plasmids, or other types of vectors. The oligonucleotides of the invention, however, may include any of the oligonucleotides sharing portions of the sequence of the 121 base sequence. Thus, oligonucleotides of the invention for each of the depicted targets may be 18, 19, 20 up to about 121 nucleotides in length. Sequence may be added non-symmetrically.

[0113] All oligonucleotides are presented, per convention, in the 5′ to 3′ orientation. The nucleotide that effects the change in the genome is underlined and presented in bold.

[0114] The first of the four oligonucleotides for each mutation is a 121 nt oligonucleotide centered about the repair/altering nucleotide. The second oligonucleotide, its reverse complement, targets the opposite strand of the DNA duplex for repair/alteration. The third oligonucleotide is the minimal 17 nt domain of the first oligonucleotide, also centered about the repair/alteration nucleotide. The fourth oligonucleotide is the reverse complement of the third, and thus represents the minimal 17 nt domain of the second.

[0115] The third column of each table presents the SEQ ID NO: of the respective repair oligonucleotide.

EXAMPLE 5

Engineering Herbicide Resistant Plants

[0116] Chemical weed control is an important tool of modern agriculture and many herbicides have been developed for this purpose. Their use has resulted in substantial increases in the yields of many crops, including, for example, maize, soybeans, and cotton. Thus while the use of fertilizers and new high-yielding crop varieties have contributed greatly to the “green revolution,” chemical weed control has also been at the forefront of technological achievement.

[0117] Herbicides having broad-spectrum activity are particularly useful because they obviate the need for multiple herbicides targeting different classes of weeds. The problem with such herbicides is that they typically also affect crops which are exposed to the herbicide. One way to overcome this is to generate plants which are resistant to one or more broad-spectrum herbicides. Such herbicide-tolerant plants may reduce the need for tillage to control weeds, thereby effectively reducing soil erosion and can reduce the quantity and number of different herbicides applied in the field.

[0118] Common herbicides used, for example, include those that inhibit the enzyme 5-enolpyruvyl-3-phosphoshikimic acid synthase (EPSPS), for example N-phosphonomethyl-glycine (e.g. glyphosate), those that inhibit acetolactate synthase (ALS) activity, for example the sulfonylureas and related herbicides, and those that inhibit dihydropteroate synthase, for example methyl[(4-amino-phenyl)sulfonyl]carbamate (e.g. Asulam). Herbicide-tolerant plants can be produced by several methods, including, for example, introducing into the genome of the plant the ability to degrade the herbicide, the capacity to produce a higher level of the targeted enzyme, and/or expressing an herbicide-tolerant allele of the enzyme.

[0119] The attached tables disclose exemplary oligonucleotides base sequences which can be used to generate site-specific mutations in plant genes that confer herbicide resistance.

12TABLE 10Genome-Altering Oligos Conferring Glyphosate ResistancePhenotype, Gene,Plant & TargetedSEQ IDAlterationAltering OligosNO:Glyphosate ResistanceAAGCGTCGGAGATTGTACTTCAACCCATTTAGAGAAATCTCCGGTC1EPSPSTTATTAAGCTTCCTGCCTCCAAGTCTCTATCAAATCGGATCCTGCArabidopsis thalianaTTCTCGCTGCTCTGTCTGAGGTATATATCACGly97AlaGTGATATATACCTCAGACAGAGCAGCGAGAAGCAGGATCCGATT2GGC-GCCTGATAGAGACTTGGAGGCAGGAAGCTTAATAAGACCGGAGATTTCTCTAATGGGTTGAAGTACAATCTCCGACGCTTGCTTCCTGCCTCCAAGT3ACTTGGAGGCAGGAAGC4Glyphosate ResistanceAAGCTTCAGAGATTGTGCTTCAACCAATCAGAGAAATCTCGGGTC5EPSPSTCATTAAGCTACCCGCATCCAAATCTCTCTCCAATCGGATCCTCCBrassica napusTTCTTGCCGCTCTATCTGAGGTACATATACTGly93AIaAGTATATGTACCTCAGATAGAGCGGCAAGAAGGAGGATCCGATT6GGA-GCAGGAGAGAGATTTGGATGCGGGTAGCTTAATGAGACCCGAGATTTCTCTGATTGGTTGAAGCACAATCTCTGAAGCTTGCTACCCGCATCCAAAT7ATTIGGATGCGGGTAGC8Glyphosate ResistanceAGCCCAACGAGATTGTGCTGCAACCCATCAAAGATATATCAGGC9EPSPS 1ACTGTTAAATTGCCTGCTTCTAAATCCCTTTCCAATCGTATTCTCCNicotiana tabacumTTCTTGCTGCCCTTTCTAAGGGAAGGACTGTGly95AlaACAGTCCTTCCCTTAGAAAGGGCAGCAAGAAGGAGAATACGATT10GGT-GCTGGAAAGGGATTTAGAAGCAGGCAATTTAACAGTGCCTGATATATCTTTGATGGGTTGCAGCACAATCTCGTIGGGCTATTGCCTGCTTCTAAAT11ATTTAGAAGCAGGCAAT12Glyphosate ResistanceATTGTTTCCTTGGTACGAAATGTCCTCCTGTTCGAATTGTCAGCA13EPSPS 2AGGGAGGCCTTCCCGCAGGGAAGGTAAAGCTCTCTGGATCAATTNicotiana tabacumAGCAGCCAGTACTTGACTGCTCTGCTTATGGCGly62AlaGCCATAAGCAGAGCAGTCAAGTACTGGCTGCTAATTGATCCAGA14GGA-GCAGAGCTTTACCTTCCCTGCGGGAAGGCCTCCCTTGCTGACAATTCGAACAGGAGGACATTTCGTACCAAGGAAACAATCCTTCCCGCAGGGAAGG15CCTTCCCGCGGGAAGG16Glyphosate ResistanceATTGTTTCCTTGGCACTGACTGGCCACCTGTTCGTGTCAATGGAA17EPSPSTCGGAGGGCTACCTGCTGGCAAGGTCAAGCTGTCTGGCTCCATCZea maysAGCAGTCAGTACTTGAGTGCCTTGCTGATGGCGly168AlaGCCATCAGCAAGGCACTCAAGTACTGACTGCTGATGGAGCCAGA18GGT-GCTCAGCTTGACCTTGCCAGCAGGTAGCCCTCCGATTCCATTGACACGAACAGGTGGGCAGTCAGTGCCAAGGAAACAATGCTACCTGCTGGCAAGG19CCTTGCCAGCAGGTAGC20Glyphosate ResistanceACTGTTTCCTTGGCACTGAATGCCCACCTGTTCGTGTCAAGGGA21EPSPSATTGGAGGACTTCCTGCTGGCAAGGTTAAGCTCTCTGGTTCCATCryza sativaCAGCAGTCAGTACTTGAGTGCCTTGCTGATGGCGly115AlaGCCATCAGCAAGGCACTCAAGTACTGACTGCTGATGGAACCAGA22GGT-GCTGAGCTTAACCTTGCCAGCAGGAAGTCCTCCAATTCCCTTGACACGAACAGGTGGGCATTCAGTGCCAAGGAAACAGTACTTCCTGCTGGCAAGG23CCTTGCCAGCAGGAAGT24Glyphosate ResistanceAGCCTTCTGAGATAGTGTTGCAACCCATTAAAGAGATTTCAGGCA25EPSPSCTGTTAAATTGCCTGCCTCTAAATCATTATCTAATAGAATTCTCCTPetunia x hybridaTCTTGCTGCCTTATCTGAAGGMCAACTGTGly93AlaACAGTTGTTCCTTCAGATAAGGCAGCAAGAAGGAGAATTCTATTA26GGC-GCCGATAATGATTTAGAGGCAGGCAATTTAACAGTGCCTGAAATCTCTTTAATGGGTTGCAACACTATCTCAGAAGGCTATTGCCTGCCTCTAAAT27ATTTAGAGGCAGGCAAT28Glyphosate ResistanceAACCCCATGAGATTGTGCTAGNACCCATCAAAGATATATCTGGTA29EPSPSCTGTTAAATTACCCGCTTCGAAATCCCTTTCCAATCGTATTCTCCTLycopersiconTCTTGCTGCCCTTTCTGAGGGAAGGACTGTesculentumACAGTCCTTCCCTCAGAAAGGGCAGCAAGAAGGAGAATACGATT30Gly97AlaGGAAAGGGATTTCGAAGCGGGTAATTTAACAGTACCAGATATATCGGT-GCTTTTGATGGGTNCTAGCACAATCTGATGGGGTTATTACCCGCTTCGAAAT31ATTTCGAAGCGGGTAAT32Glyphosate ResistanceATTGTTTCCTTGGCACTGACTGCCCACCTGTTCGKATCAACGGGA33EPSPSTTGGAGGGCTACCTGCTGGCAAGGTTAAGCTGTCTGGTTCCAITLolium rigidumAGCAGCCAATACTTGAGTTCCTTGCTGATGGCGly107AlaGCCATCAGCAAGGAACTCAAGTATTGGCTGCTGATGGAACCAGA34GGT-GCTCAGCTTAACCTTGCCAGCAGGTAGCCCTCCAATGCCGTTGATCGAACAGGTGGGCAGTCAGTGCCAAGGAAACAATGCTACCTGCTGGCAAGG35CCTTGCCAGCAGGTAGC36

[0120]

13

TABLE 11

Genome-Altering Oligos Conferring Imidazolinone

and Sulfonylurea Herbicide Resistance

Phenotype, Gene,

Plant & Targeted

SEQ ID

Alteration
Altering Oligos
NO:

Sulfonylurea
AGCGGATTAGCCGATGCGTTGTTAGATAGTGTTCCTCTTGTAGCA
37

Resistance
ATCACAGGACAAGTCTCTCGTCGTATGATTGGTACAGATGCGTTT

ALS
CAAGAGACTCCGATTGTTGAGGTAACGCGTT

Arabidopsis thaliana
AACGCGTTACCTCAACAATCGGAGTCTCTTGAAACGCATCTGTAC
38

Pro197Ser
CAATCATACGACGAGAGACTTGTCCTGTGATTGCTACAAGAGGAA

CCT-TCT
CACTATCTAACAACGCATCGGCTAATCCGCT

GACAAGTCTCTCGTCGT
39

ACGACGAGAGACTTGTC
40

Sulfonylurea
AGCGGATTAGCCGATGCGTTGTTAGATAGTGTTCCTCTTGTAGCA
41

Resistance
ATCACAGGACAAGTCCAGCGTCGTATGATTGGTACAGATGCGTTT

ALS
CAAGAGACTCCGATTGTTGAGGTAACGCGTT

Arabidopsis thaliana

AACGCGTTACCTCAACAATCGGAGTCTCTTGAAACGCATCTGTAC
42

Pro197GLN
CAATCATACGACGCTGGACTTGTCCTGTGATTGCTACAAGAGGAA

CCT-CAG
CACTATCTAACAACGCATCGGCTAATCCGCT

ACAAGTCCAGCGTCGTC
43

TACGACGCTGGACTTGT
44

Sulfonylurea
AGCGGATTAGCCGATGCGTTGTTAGATAGTGTTCCTCTTGTAGCA
45

Resistance
ATCACAGGACAAGTCCAACGTCGTATGATTGGTACAGATGCGTTT

ALS
CAAGAGACTCCGATTGTTGAGGTAACGCGTT

Arabidopsis thaliana

AACGCGTTACCTCAACAATCGGAGTCTCTTGAAACGCATCTGTAC
46

Pro197GLN
CAATCATACGACGTTGGACTTGTCCTGTGATTGCTACAAGAGGAA

CCT-CAA
CACTATCTAACAACGCATCGGCTAATCCGCT

ACAAGTCCAACGTCGTA
47

TACGACGTTGGACTTGT
48

Imidazolinone
GACCTTACCTGTTGGATGTGATTTGTCCGCACCAAGAACATGTGT
49

Resistance
TGCCGATGATCCCGAACGGTGGCACTTTCAACGATGTCATAACGG

ALS
AAGGAGATGGCCGGATTAAATACTGAGAGAT

Arabidopsis thaliana

ATCTCTCAGTATTTAATCCGGCCATCTCCTTCCGTTATGACATCGT
50

Ser653Asn
TGAAAGTGCCACCGTTCGGGATCATCGGCAACACATGTTCTTGGT

AGT-AAC
GCGGACAAATCACATCCAACAGGTAAGGTC

GATCCCGAACGGTGGCA
51

TGCCACCGTTCGGGATC
52

Imidazolinone
GACCTTACCTGTTGGATGTGATTTGTCCGCACCAAGAACATGTGT
53

Resistance
TGCCGATGATCCCGAATGGTGGCACTTTCAACGATGTCATAACGG

ALS
AAGGAGATGGCCGGATTAAATACTGAGAGAT

Arabidopsis thaliana

ATCTCTCAGTATTTAATCCGGCCATCTCCTTCCGTTATGACATCGT
54

Ser653Asn
TGAAAGTGCCACCATTCGGGATCATCGGCAACACATGTTCTTGGT

AGT-AAT
GCGGACAAATCACATCCAACAGGTAAGGTC

GATCCCGAATGGTGGCA
55

TGCCACCATTCGGGATC
56

Sulfonylurea
TCCGCGCTCGCCGACGCGCTGCTCGACTCCGTCCCGATGGTCGC
57

Resistance
CATCACGGGCCAGGTCTCCCGCCGCATGATCGGCACCGACGCCT

ALS
TCCAGGAGACGCCCATAGTCGAGGTCACCCGCT

Oryza saliva

AGCGGGTGACCTCGACTATGGGCGTCTCCTGGAAGGCGTCGGTG
58

Pro171Ser
CCGATCATGCGGCGGGAGACCTGGCCCGTGATGGCGACCATCG

CCC-TCC
GGACGGAGTCGAGCAGCGCGTCGGCGAGCGCGGA

GCCAGGTCTCCCGCCGC
59

GCGGCGGGAGACCTGGC
60

Sulfonylurea
CCGCGCTCGCCGACGCGCTGCTCGACTCCGTCCCGATGGTCGCC
61

Resistance
ATCACGGGCCAGGTCCAACGCCGCATGATCGGCACCGACGCCTT

ALS
CCAGGAGACGCCCATAGTCGAGGTCACCCGCTC

Oryza saliva

GAGCGGGTGACCTCGACTATGGGCGTCTCCTGGAAGGCGTCGGT 62

Pro171Gln
GCCGATCATGCGGCGTTGGACCTGGCCCGTGATGGCGACCATCG

CCC-CAA
GGACGGAGTCGAGCAGCGCGTCGGCGAGCGCGG

CCAGGTCCAACGCCGCA
63

TGCGGCGTTGGACCTGG
64

Sulfonylurea
CCGCGCTCGCCGACGCGCTGCTCGACTCCGTCCCGATGGTCGCC
65

Resistance
ATCACGGGCCAGGTCCAGCGCCGCATGATCGGCACCGACGCCTT

ALS
CCAGGAGACGCCCATAGTCGAGGTCACCCGCTC

Oryza saliva

GAGCGGGTGACCTCGACTATGGGCGTCTCCTGGAAGGCGTCGGT
66

Pro171Gln
GCCGATCATGCGGCGCTGGACCTGGCCCGTGATGGCGACCATCG

CCC-CAG
GGACGGAGTCGAGCAGCGCGTCGGCGAGCGCGG

CCAGGTCCAGCGCCGCA
67

TGCGGCGCTGGACCTGG
68

Imidazolinone
GGCCATACTTGTTGGATATCATCGTCCCGCACCAGGAGCATGTGC
69

Resistance
TGCCTATGATCCCAAATGGGGGCGCATTCAAGGACATGATCCTGG

ALS
ATGGTGATGGCAGGACTGTGTATTAATCTAT

Oryza saliva

ATAGATTAATACACAGTCCTGCGATCACCATCCAGGATCATGTCCT
70

Ilee627Asn
TGAATGCGCCCCCATTTGGGATCATAGGCAGCACATGCTCCTGGT

ATT-AAT
GCGGGACGATGATATCCAACAAGTATGGCC

GATCCCAAATGGGGGCG
71

CGCCCCCATTTGGGATC
72

Sulfonylurea
TCCGCGCTCGCCGACGCGCTGCTCGATTCCGTCCCCATGGTCGC
73

Resistance
CATCACGGGACAGGTGTCGCGACGCATGATTGGCACCGACGCCT

ALS
TCCAGGAGACGCCCATCGTCGAGGTCACCCGCT

Zea mays

AGCGGGTGACCTCGACGATGGGCGTCTCCTGGAAGGCGTCGGT
74

Pro165Ser
GCCAATCATGCGTCGCGACACCTGTCCCGTGATGGCGACCATGG

CCG-TCG
GGACGGAATCGAGCAGCGCGTCGGCGAGCGCGGA

GACAGGTGTCGCGACGC
75

GCGTCGCGACACCTGTC
76

Sulfonylurea
CCGCGCTCGCCGACGCGCTGCTCGATTCCGTCCCCATGGTCGCC
77

Resistance
ATCACGGGACAGGTGCAGCGACGCATGATTGGCACCGACGCCTT

ALS
CCAGGAGACGCCCATCGTCGAGGTCACCCGCTC

Zea mays

GAGCGGGTGACCTCGACGATGGGCGTCTCCTGGAAGGCGTCGG
78

Pro165Gln
TGCCAATCATGCGTCGCTGCACCTGTCCCGTGATGGCGACCATG

CCG-CAG
GGGACGGAATCGAGCAGCGCGTCGGCGAGCGCGG

ACAGGTGCAGCGACGCA
79

TGCGTCGCTGCACCTGT
80

Imidazolinone
GGCCGTACCTCTTGGATATAATCGTCCCACACCAGGAGCATGTGT
81

Resistance
TGCCTATGATCCCTAATGGTGGGGCTTTCAAGGATATGATCCTGG

ALS
ATGGTGATGGCAGGACTGTGTACTGATCTAA

Zea mays

TTAGATCAGTACACAGTCCTGCCATCACCATCCAGGATCATATCCT
82

Ser621Asn
TGAAAGCCCCACCATTAGGGATCATAGGCAACACATGCTCCTGGT

AGT-AAT
GTGGGACGATTATATCCAAGAGGTACGGCC

GATCCCTAATGGTGGGG
83

CCCCACCATTAGGGATC
84

Imidazolinone
GGCCGTACCTCTTGGATATAATCGTCCCACACCAGGAGCATGTGT
85

Resistance
TGCCTATGATCCCTAACGGTGGGGCTTTCAAGGATATGATCCTGG

ALS
ATGGTGATGGCAGGACTGTGTACTGATCTAA

Zea mays

TTAGATCAGTACACAGTCCTGCCATCACCATCCAGGATCATATCCT
86

Ser621Asn
TGAAAGCCCCACCGTTAGGGATCATAGGCAACACATGCTCCTGGT

AGT-AAC
GTGGGACGATTATATCCAAGAGGTACGGCC

GATCCCTAACGGTGGGG
87

CCCCACCGTTAGGGATC
88

Sulfonylurea
TCCGCGCTCGCCGACGCCGTCCTCGACTCCATCCCCATGGTGGC
89

Resistance
CATCACGGGGCAGGTCTCGCGCCGCATGATCGGCACGGACGCCT

ALS
TCCAGGAGACGCCCATCGTCGAGGTCACCCGCT

Lolium multiflorum

AGCGGGTGACCTCGACGATGGGCGTCTCCTGGAAGGCGTCCGTG
90

Pro167Ser
CCGATCATGCGGCGCGAGACCTGCCCCGTGATGGCCACCATGG

CCG-TCC
GGATGGAGTVGAGGAGGGCCTCGGCGACCCCCCA

GGCAGGTCTCGCGCCGC
91

GCGGCGCGAGACCTGCC
92

Sulfonylurea
CCGCGCTCGCCGACGCCCTCCTCGACTCCATCCCCATGGTGGCC
93

Resistance
ATCACGGGGCAGGTCCAGCGCCGCATGATCGGCACGGACGCCTT

ALS
CCAGGAGACGCCCATCGTCGAGGTCACCCGCTC

Lolium multiflorum

GAGCGGGTGACCTCGACGATGGGCGTCTCCTGGAAGGCGTCCGT
94

Pro167Gln
GCCGATCATGCGGCGCTGGACCTGCCCCGTGATGGCCACCATGG

CCG-CAG
GGATGGAGTCGAGGAGGGCGTCGGCGAGCGCGG

GCAGGTCCAGCGCCGCA
95

TGCGGCGCTGGACCTGC
96

Imidazolinone
CTGGGCCATACTTGTTGGATATCATCGTCCCTCACCAGGAGCATG
97

Resistance
TGCTGCCTATGATCCCTAACGGTGGTGCTTTCAAGGACATTATCA

ALS
TGGAAGGTGATGGCAGGATTTCGTATTAAAC

Lolium multiflorum

GTTTAATACGAAATCCTGCCATCACCTTCCATGATAATGTCGTTGA
98

Ser623Asn
AAGCACCACCGTTAGGGATCATAGGCAGCACATGCTCCTGGTGA

AGC-AAC
GGGACGATGATATCCAACAAGTATGGCCCAG

GATCCCTAACGGTGGTG
99

CACCACCGTTAGGGATC
100

Sulfonylurea
TCCGCGCTCGCCGACGGTCTCCTCGACTCCATCGCCATGGTCGC
101

Resistance
CATCACGGGCCAGGTCTCACGCCGCATGATCGGCACGGACGCGT

ALS
TCCAGGAGACGCCCATAGTGGAGGTCACGCGCT

Hordeum vulgare

AGCGCGTGACCTCCACTATGGGCGTCTCCTGGAACGCGTCCGTG
102

Pro68Ser
CGGATCATGCGGCGTGAGACCTGGCCCGTGATGGCGACCATGG

CCA-TCA
GGATGGAGTCGAGGAGAGCGTCGGCGAGCGCGGA

GCCAGGTCTCACGCCGC
103

GCGGCGTGAGACCTGGC
104

Sulfonyurea
CCGCGCTCGCCGACGCTCTCCTCGACTCCATCCCCATGGTCGCC
105

Resistance
ATCACGGGCCAGGTCCAACGCCGCATGATCGGCACGGACGCGTT

ALS
CCAGGAGACGCCCATAGTGGAGGTCACGCGCTC

Hordeum vulgare

GAGCGCGTGACCTCCACTATGGGCGTCTCCTGGAACGCGTCCGT
106

Pro68Gln
GCCGATCATGCGGCGTTGGACCTGGCCCGTGATGGCGACCATGG

CCA-CAA
GGATGGAGTCGAGGAGAGCGTCGGCGAGCGCGG

CCAGGTCCAACGCCGCA
107

TGCGGCGTTGGACCTGG
108

Imidazolinone
CCCAGGGCCGTACCTGCTGGATATCATTGTCCCGCATCAGGAGC
109

Resistance
ACGTGCTGCCTATGATCCCAAACGGTGGTGCTTTCAAGGACATGA

ALS
TCATGGAGGGTGATGGCAGGACCTCGTACTGA

Hordeum vulgare

TCAGTACGAGGTCCTGCCATTCACCCTCCATGATCATGTCCTTGAA
110

Ser524Asn
AGCACCACCGTTTGGGATCATAGGCAGCACGTGCTCCTGATGCG

AGC-AAC
GGACAATGATATCCAGCAGGTACGGCCCTGGG

GATCCCAAACGGTGGTG
111

CACCACCGTTTGGGATC
112

Sulfonylurea
AGTGGTCTCGCTGATGCAATGCTCGATAGTATCCCTCTCGTGGCG
113

Resistance
ATCACTGGTCAAGTCTCTCGTCGGATGATCGGTACCGATGCTTTC

ALS
CAGGAAACTCCAATTGTTGAGGTAACAAGGT

Gossypium hirsutum

ACCTTGTTACCTCAACAATTGGAGTTTCCTGGAAAGCATCGGTAC
114

Pro186Ser
CGATCATCCGACGAGAGACTTGACCAGTGATCGCCACGAGAGGG

CCT-TCT
ATACTATCGAGCATTGCATCAGCGAGACCACT

GTCAAGTCTCTCGTCGG
115

CCGACGAGAGACTTGAC
116

Sulfonylurea
GTGGTCTCGCTGATGCAATGGTCGATAGTATCCCTCTCGTGGCGA
117

Resistance
TCACTGGTCAAGTCCAACGTCGGATGATCGGTACCGATGCTTTCC

ALS
AGGAAACTCCAATTGTTGAGGTAACAAGGTC

Gossypium hirsutum

GACCTTGTTACCTCAACAATTGGAGTTICCTGGAAAGCATCGGTA
118

Pro186Gln
CCGATCATCCGACGTTGGACTTGACCAGTGATCGCCACGAGAGG

CCT-CAA
GATACTATCGAGCATTGCATCAGCGAGACCAC

TCAAGTCCAACGTCGGA
119

TCCGACGTTGGACTTGA
120

Sulfonylurea
GTGGTCTCGCTGATGCAATGCTCGATAGTATCCCTCTCGTGGCGA
121

Resistance
TCACIGGTCAAGTCCAGCGTCGGATGATCGGTACCGATGCTTTCC

ALS
AGGAAACTCCAATTGTTGAGGTAACAAGGTC

Gossypium hirsutum

GACCTTGTTACCTCAACAATTGGAGTTTCCTGGAAAGCATCGGTA
122

Pro186Gln
CCGATCATCCGACGCTGGACTTGACCAGTGATCGCCACGAGAGG

CCT-CAG
GATACTATCGAGCATTGCATCAGCGAGACCAC

TCAAGTCCAGCGTCGGA
123

TCCGACGCTGGACTTGA
124

Imidazolinone
GACCTTACTTGTTGGATGTGATTGTCCCACATCAAGAACATGTCCT
125

Resistance
GCCTATGATCCCCAATGGAGGCGCTTTCAAAGATGTGATCACAGA

ALS
GGGTGATGGAAGAACACAATATTGACCTCA

Gossypium hirsutum

TGAGGTCAATATTGTGTTCTTCCATCACCCTCTGTGATCACATCTT
126

Ser642Asn
TGAAAGCGCCTCCATTGGGGATCATAGGCAGGACATGTTCTTGAT

AGT-AAT
GTGGGACAATCACATCCAACAAGTAAGGTC

GATCCCCAATGGAGGCG
127

CGCCTCCATTGGGGATC
128

Sulfonylurea
TCTGGTCTTGCTGATGCACTTCTTGACTCAGTCCCTCTTGTCGCCA
129

Resistance
TTACTGGGCAAGTTTCCCGGCGTATGATTGGTACTGATGCTTTTCA

ALS
AGAGACTCCAATTGTTGAGGTAACTCGAT

Amaranthus

ATCGAGTTACCTCAACAATTGGAGTCTCTTGAAAAGCATCAGTACC
130

retroflexus

AATCATACGCCGGGAAACTTGCCCAGTAATGGCGACAAGAGGGA

Pro192Ser
CTGAGTCAAGAAGTGCATCAGCAAGACCAGA

CCC-TCC
GGCAAGTTTCCCGGCGT
131

ACGCCGGGAAAGTTGCC
132

Sulfonylurea
CTGGTCTTGCTGATGCACTTCTTGACTCAGTCCCTCTTGTCGCCAT
133

Resistance
TACTGGGCAAGTTCAACGGCGTATGATTGGTACTGATGCTTTTCA

ALS
AGAGACTCCAATTGTTGAGGTAACTCGATC

Amaranthus

GATCGAGTTACCTCAACAATTGGAGTCTCTTGAAAAGCATCAGTAC
134

retroflexus

CAATCATACGCCGTTGAACTTGCCCAGTAATGGCGACAAGAGGGA

Pro192Gln
CTGAGTCAAGAAGTGCATCAGCAAGACCAG

CCC-CAA
GCAAGTTCAACGGCGTA
135

TACGCCGTTGAACTTGC
136

Sulfonylurea
CTGGTCTTGCTGATGCACTTCTTGACTCAGTCCCTCTTGTCGCCAT
137

Resistance
TACTGGGCAAGtTCAGCGGCGTATGATTGGTACTGATGCTTTTCA

ALS
AGAGACTCCAATTGTTGAGGTAACTCGATC

Amaranthus

GATCGAGTTACCTCAACAATTGGAGTCTCTTGAAAAGCATCAGTAC
138

retroflexus

CAATCATACGCCGCTGAACTTGCCCAGTAATGGCGACAAGAGGG

Pro192Gln
ACTGAGTCAAGAAGTGCATCAGCAAGACCAG

CCC-CAG
GCAAGTTCAGCGGCGTA
139

TACGCCGCTGAACTTGC
140

Imidazolinone
GACCGTATCTTGCTGGATGTTAATCGTACCACATCAGGAGCATGTGC
141

Resistance
TGCCTAIGATCCCTAACGGTGCCGCCTTCAAGGACACCATAACAG

ALS
AGGGTGATGGAAGAAGGGGTTATTAGTTGGT

Amaranthus

ACCAACTAATAAGCCCTTCTTCCATTCACCCTCTGTTATGGTGTCCT
142

retroflexus

TGAAGGCGGCACCGTTAGGGATCATAGGCAGCACATGCTCCTGA

Ser652Asn
TGTGGTACGATTACATCCAGCAGATACGGTC

AGC-AAC
GATCCCTAACGGTGCCG
143

CGGCACCGTTAGGGATC
144

Sulfonylurea
AGCGGCCTCGCTGACGCGCTACTGGATAGCGTCCCCATTGTTGC
145

Resistance
TATAACAGGTCAAGTGTCACGTAGGATGATAGGTACTGATGCTTTT

ALS 1
CAGGAAACTCCTATTGTITGAGGTAACTAGAT

Nicotiana tabacum

ATCTAGTTACCTCAACAATAGGAGTTTCCTGAAAAGCATCAGTACC
146

Pro194Ser
TATCATCCTACGTGACACTTGACCTGTTATAGCAACAATGGGGAC

CCA-TCA
GCTATCCAGTAGCGCGTCAGCGAGGCCGCT

GTCAAGTGTCACGTAGG
147

CCTACGTGACACTTGAC
148

Sulfonylurea
GCGGCCTCGCTGACGCGCTACTGGATAGCGTCCCCATTGTTGCT
149

Resistance
ATAACAGGTCAAGTGCAACGTAGGATGATAGGTACTGATGCTTTT

ALS 1
CAGGAAACTCCTATTGTTGAGGTAACTAGATC

Nicotiana tabacum

GATCTAGTTACCTCAACAATAGGAGTTTCCTGAAAAGCATCAGTAC
150

Pro194Gln
CTATCATCCTACGTTGCACTTGACCTGTTATAGCAACAATGGGGA

CCA-CAA
CGCTATCCAGTAGCGCGTCAGCGAGGCCGC

TCAAGTGCAACGTAGGA
151

TCCTACGTTGCACTTGA
152

Imidazolinone
GGCCATACTTGTTGGATGTGATTGTACCTCATCAGGAACATGTTTT
153

Resistance
ACCTATGATTCCCAATGGCGGAGCTTTCAAAGATGTGATCACAGA

ALS 1
GGGTGACGGGAGAAGTTCCTATTGAGTTTG

Nicotiana tabacum

CAAACTGAATAGGAACTTCTCCCGTCACCCTCTGTGATCACATCTT
154

Ser650Asn
TGAAAGCTCCGCCATTGGGAATCATAGGTAAAACATGTTCCTGAT

AGT-AAT
GAGGTACAATCACATCCAACAAGTATGGCC

GATTCCCAATGGCGGAG
155

CTCCGCCATTGGGAATC
156

Sulfonylurea
AGTGGCCTCGCGGACGCCCTACTGGATAGCGTCCCCATTGTTGC
157

Resistance
TATAACCGGTCAAGTGTCACGTAGGATGATCGGTACTGATGCTTT

ALS 2
TCAGGAAACTCCGATTGTTGAGGTAACTAGAT

Nicotiana tabacum

ATCTAGTTACCTCAACAATCGGAGTTTCCTGAAAAGCATCAGTACC
158

Pro191Ser
GATCATCCTACGTGACACTTGACCGGTTATAGCAACAATGGGGAC

CCA-TCA
GCTATCCAGTAGGGCGTCCGCGAGGCCACT

GICAAGTGTCACGTAGG
159

CCTACGTGACACTTGAC
160

Sulfonylurea
GTGGCCTCGCGGACGCCCTACTGGATAGCGTCCCCATTGTTGCT
161

Resistance
ATAACCGGTCAAGTGCAACGTAGGATGATCGGTACTGATGCTTTT

ALS 2
CAGGAAACTCCGATTGTTGAGGTAACTAGATC

Nicotiana tabacum

GATCTAGTTACCTCAACAATCGGAGTTTCCTGAAAAGCATCAGTAC
162

Pro191Gln
CGATCATCCTACGTTGCACTTGACCGGTTATAGCAACAATGGGGA

CCA-CAA
CGCTATCCAGTAGGGCGTCCGCGAGGCCAC

TCAAGTGCAACGTAGGA
163

TCCTACGTTGCACTTGA
164

Imidazolinone
GGCCATACTTGTTGGATGTGATTGTACCTCATCAGGAACATGTTCT
165

Resistance
ACCTATGATTCCCAATGGCGGGGCTTTCAAAGATGTGATCACAGA

ALS 2
GGGTGACGGGAGAAGTTCCTATTGACTTTG

Nicotiana tabacum

CAAAGTCAATAGGAACTTCTCCCGTCACCCTCTGTGATCACATCTT
166

Ser647Asn
TGAAAGCCCCGCCATTGGGAATCATAGGTAGAACATGTTCCTGAT

AGT-AAT
GAGGTACAATCACATCCAACAAGTATGGCC

GATTCCCAATGGCGGGG
167

CCCCGCCATTGGGAATC
168

Sulfonylurea
AGTGGTCTTGCTGATGCTTTATTAGACAGTGTTCCAATGGTTGCTA
169

Resistance
TTACTGGTCAAGTTTCCAGGAGAATGATTGGAACAGATGCGTTTC

ALS
AAGAAACCCCTATTGTTGAGGTAACACGTT

Xanthium spp.
AACGTGTTACCTCAACAATAGGGGTTTCTTGAAACGCATCTGTTCC
170

Pro175Ser
AATCATTCTCCTGGAAACTTGACCAGTAATAGCAACCATTGGAACA

CCC-TCC
CTGTCTAATAAAGCATCAGCAAGACCACT

GTCAAGTTTCCAGGAGA
171

TCTCCTGGAAACTTGAC
172

Sulfonylurea
GTGGTCTTGCTGATGCTTTATTAGACAGTGTTCCAATGGTTGCTAT
173

Resistance
TACTGGTCAAGTTCAAAGGAGAATGATTGGAACAGATGCGTTTCA

ALS
AGAAACCCCTATTGTTGAGGTAACACGTTC

Xanthium spp.
GAACGTGTTACCTCAACAATAGGGGTTTCTTGAAACGCATCTGTTC
174

Pro175Gln
CAATCATTCTCCTTTGAACTTGACCAGTAATAGCAACCATTGGAAC

CCC-CAA
ACTGTCTAATAAAGCATCAGCAAGACCAC

TCAAGTTCAAAGGAGAA
175

TTCTCCTTTGAACTTGA
176

Sulfonylurea
GTGGTCTTGCTGATGCTTTATTAGACAGTGTTCCAATGGTTGCTAT
177

Resistance
TACTGGTCAAGTTCAGAGGAGAATGATTGGAACAGATGCGTTTCA

ALS
AGAAACCCCTATTGTTGAGGTAACACGTTC

Xanthium spp.
GAACGTGTTACCTCAACAATAGGGGTTTCTTGAAACGCATCTGTTC 178

Pro175Gln
CAATCATTCTCCTCTGAACTTGACCAGTAATAGCAACCATTGGAAC

CCC-CAG
ACTGTCTAATAAAGCATCAGCAAGACCAC

TCAAGTTCAGAGGAGAA
179

TTCTCCTCTGAACTTGA
180

Imidazolinone
GGGCCTTACTTGTTGGATGTGATCGTGCCCCATCAAGAACATGTG
181

Resistance
TTGCCCATGATCCCGAATGGTGGAGGTTTCATGGATGTGATCACC

ALS
GAAGGCGACGGCAGAATGAAATATTGAGCTT

Xanthium spp.
AAGCTCAATATTTCATTCTGCCGTCGCCTTCGGTGATCACATCCAT
182

Ala631Asn
GAAACCTCCACCATTCGGGATCATGGGCAACACATGTTCTTGATG

GCT-AAT
GGGCACGATCACATCCAACAAGTAAGGCCC

TGATCCCGAATGGTGGA
183

TCCACCATTCGGGATCA
184

Sulfonylurea
TCCGGGTTTGCTGATGCTTTGCTCGATTCCGTTCCACTGGTGGCG
185

Resistance
ATCACGGGGCAGGTGTCGCGGCGAATGATTGGGACGGATGCTTT

ALS
TCAGGAGACTCCTATTGTTGAGGTAACACGGT

Bassia scoparia

ACCGTGTTACCTCAACAATAGGAGTCTCCTGAAAAGCATCCGTCC
186

Pro189Ser
CAATCATTCGCCGCGACACCTGCCCCGTGATCGCCACCAGTGGA

CCG-TCG
ACGGAATCGAGCAAAGCATCAGCAAACCCGGA

GGCAGGTGTCGCGGCGA
187

TCGCCGCGACACCTGCC
188

Sulfonylurea
CCGGGTTTGGTGATGCTTTGCTCGATTCCGTTCCACTGGTGGCGA
189

Resistance
TCACGGGGCAGGTGCAGCGGCGAATGATTGGGACGGATGCTTTT

ALS
CAGGAGACTCCTATTGTTGAGGTAACACGGTC

Bassia scoparia

GACCGTGTTACCTCAACAATAGGAGTCTCCTGAAAAGCATCCGTC
190

Pro189Gln
CCAATCATTCGCCGCTGCACCTGCCCCGTGATCGCCACCAGTGG

CCG-CAG
AACGGAATCGAGCAAAGCATCAGCAAACCCGG

GCAGGTGCAGCGGCGAA
191

TTCGCCGCTGCAGCTGC
192

Imidazolinone
GACCTTACCTGCTTGATGTGATTGTACCTCATCAGGAGCATGTGC
193

Resistance
TGCCTATGATTCCTAATGGTGCAGCCTTCAAGGATATCATTAACGA

ALS
AGGTGATGGAAGAACAAGTTATTGATGTTC

Bassia scoparia

GAACATCAATAACTTGTTCTTCCATCACCTTCGTTAATGATATCCTT
194

Ser649Asn
GAAGGCTGCACCATTAGGAATCATAGGCAGCACATGCTCCTGATG

AGT-AAT
AGGTACAATCACATCAAGCAGGTAAGGTC

GATTCGTAATGGTGCAG
195

CTGCACCATTAGGAATC
196

Sulfonylurea
AGCGGGTTAGCAGACGCGATGCTTGACAGTGTTCCTCTTGTCGCC
197

Resistance
ATTACAGGACAGGTCTCTCGCCGGATGATCGGTACTGACGCCTTC

ALS 1
CAAGAGACACCAATCGTTGAGGTAACGAGGT

Brassica napus

ACCTCGTTACCTCAACGATTGGTGTCTCTTGGAAGGCGTCAGTAC
198

Pro182Ser
CGATCATCCGGCGAGAGACCTGTCCTGTAATGGCGACAAGAGGA

CCT-TCT
ACACTGTCAAGCATCGCGTCTGCTAACCCGCT

GACAGGTCTCTCGCCGG
199

CCGGCGAGAGACCTGTC
200

Sulfonylurea
GCGGGTTAGCAGACGCGATGCTTGACAGTGTTCCTCTTGTCGCCA
201

Resistance
TTACAGGACAGGTCCAACGCCGGATGATCGGTACTGACGCCTTC

ALS 1
CAAGAGACACCAATCGTTGAGGTAACGAGGTC

Brassica napus

GACCTCGTTACCTCAACGATTGGTGTCTCTTGGAAGGCGTCAGTA
202

Pro182Gln
CCGATCATCCGGCGTTGGACCTGTCCTGTAATGGCGACAAGAGG

CCT-CAA
AACACTGTCAAGCATCGCGTCTGCTAACCCGC

ACAGGTCCAACGCCGGA
203

TCCGGCGTTGGACCTGT
204

Sulfonylurea
GCGGGTTAGCAGACGCGATGCTTGACAGTGTTCCTCTTGTCGCCA
205

Resistance
TTACAGGACAGGTCCAGCGCCGGATGATCGGTACTGACGCCTTC

ALS 1
CAAGAGACACCAATCGTTGAGGTAACGAGGTC

Brassica napus

GACCTCGTTACCTCAACGATTGGTGTCTCTTGGAAGGCGTCAGTA
206

Pro182Gln
CCGATCATCCGGCGCTGGACCTGTCCTGTAATGGCGACAAGAGG

CCT-CAG
AACACTGTCAAGCATCGCGTCTGCTAACCCGC

ACAGGTCCAGCGCCGGA
207

TCCGGCGCTGGACCTGT
208

Imidazolinone
GACCATACCTGTTGGATGTGATATGTCCGCACCAAGAACATGTGT
209

Resistance
TACCGATGATCCCAAATGGTGGCACTTTCAAAGATGTAATAACAG

ALS 1
AAGGGGATGGTCGCACTAAGTACTGAGAGAT

Brassica napus

ATCTCTCAGTACTTAGTGCGACCATCCCCTTCTGTTATTACATCTTT
210

Ser638Asn
GAAAGTGCCACCATTTGGGATCATCGGTAACACATGTTCTTGGTG

AGT-AAT
CGGACATATCACATCCAACAGGTATGGTC

GATCCCAAATGGTGGCA
211

TGCCACCATTTGGGATC
212

Sulfonylurea
CAGCGGGTTAGCAGACGCGATGCTTGACAGTGTTCCTCTTGTCGC
213

Resistance
CATTACAGGACAGGTTCCTCGCCGGATGATCGGTACTGACGCCTT

ALS 2
CCAAGAGACACCAATCGTTGAGGTAACGAGG

Brassica napus

CCTCGTTACCTCAACGATTGGTGTCTCTTGGAAGGCGTCAGTACC
214

Pro126Ser
GATCATCCGGCGAGGAACCTGTCCTGTAATGGCGACAAGAGGAA

CCC-TCC
CACTGTCAAGCATCGCGTCTGCTAACCCGCTG

GGACAGGTTCCTCGCCG
215

CGGCGAGGAACCTGTCC
216

Sulfonylurea
AGCGGGTTAGCAGACGCGATGCTTGACAGTGTTCCTCTTGTCGCC
217

Resistance
ATTACAGGACAGGTCACTCGCCGGATGATCGGTACTGACGCCTTC

ALS 2
CAAGAGACACCAATCGTTGAGGTAACGAGGT

Brassica napus

ACCTCGTTACCTCAACGATTGGTGTCTCTTGGAAGGCGTCAGTAC
218

Pro126Gln
CGATCATCCGGCGAGTGACCTGTCCTGTAATGGCGACAAGAGGA

CCC-CAG
ACACTGTCAAGCATCGCGTCTGCTAACCCGCT

GACAGGTCACTCGCCGG
219

CCGGCGAGTGACCTGTC
220

Imidazolinone
GACCATACCTGTTGGATGTGATATGTCCGCACCAAGAACATGTGT
221

Resistance
TACCGATGATCCCAAATGGTGGCACTTTCAAAGATGTAATAACAG

ALS 2
AAGGGGATGGTCGCACTAAGTACTGAGAGAT

Brassica napus

ATCTCTCAGTACTTAGTGCGACCATCCCCTTCTGTTATTACATCTTT
222

Ser582Asn
GAAAGTGCCACCATTTGGGATCATCGGTAACACATGTTCTTGGTG

AGT-AAT
CGGACATATCACATCCAACAGGTATGGTC

GATCCCAAATGGTGGCA
223

TGCCACCATTTGGGATC
224

Sulfonylurea
AGCGGGTTAGCCGACGCGATGCTTGACAGTGTTCCTCTCGTCGC
225

Resistance
CATCACAGGACAGGTCTCTCGCCGGATGATCGGTACTGACGCGT

ALS 3
TCCAAGAGACGCCAATCGTTGAGGTAACGAGGT

Brassica napus

ACCTCGTTACCTCAACGATTGGCGTCTCTTGGAACGCGTCAGTAC
226

Pro179Ser
CGATCATCCGGCGAGAGACCTGTCCTGTGATGGCGACGAGAGGA

CCT-TCT
ACACTGTCAAGCATCGCGTCGGCTAACCCGCT

GACAGGTCTCTCGCCGG
227

CCGGCGAGAGACCTGTC
228

Sulfonylurea
GCGGGTTAGCCGACGCGATGCTTGACAGTGTTCCTCTCGTCGCC
229

Resistance
ATCACAGGACAGGTCCAACGCCGGATGATCGGTACTGACGCGTT

ALS 3
CCAAGAGACGCCAATCGTTGAGGTAACGAGGTC

Brassica napus

GACCTCGTTACCTCAACGATTGGCGTCTCTTGGAACGCGTCAGTA
230

Pro179Gln
CCGATCATCCGGCGTTGGACCTGTCCTGTGATGGCGACGAGAGG

CCT-CAA
AACACTGTCAAGCATCGCGTCGGCTAACCCGC

ACAGGTCCAAee CGCCGGA
231

TCCGGCGTTGGACCTGT
232

Sulfonylurea
GCGGGTTAGCCGACGCGATGCTTGACAGTGTTCCTCTCGTCGCC
233

Resistance
ATCACAGGACAGGTCCAGCGCCGGATGATCGGTACTGACGCGTT

ALS 3
CCAAGAGACGCCAATCGTTGAGGTAACGAGGTC

Brassica napus

GACCTCGTTACCTCAACGATTGGCGTCTCTTGGAACGCGTCAGTA
234

Pro179Gln
CCGATCATCCGGCGCTGGACCTGTCCTGTGATGGCGACGAGAGG

CCT-CAG
AACACTGTCAAGCATCGCGTCGGCTAACCCGC

ACAGGTCCAGCGCCGGA
235

TCCGGCGCTGGACCTGT
236

Imidazolinone
GACCGTACCTGTTGGATGTCATCTGTCCGCACCAAGAACATGTGT
237

Resistance
TACOGATGATCCCAAATGGTGGCACTTTCAAAGATGTAATAACCG

ALS 3
AAGGGGATGGTCGCACTAAGTACTGAGAGAT

Brassica napus

ATCTCTCAGTACTTAGTGCGACCATCCCCTTCGGTTATTACATCTT
238

Ser635Asn
TGAAAGTGCCACCATTTGGGATCATCGGTAACACATGTTCTTGGT

AGT-AAT
GCGGACAGATGACATCCAACAGGTACGGTC

GATCCCAAATGGTGGCA
239

TGCCACCATTTGGGATC
240

Sultonylurea
TCCGCGCTCGCCGACGCGCTGCTCGACTCCGTCCCGATGGTCGC
241

Resistance
CATCACGGGCCAGGTCTCCCGCCGCATGATCGGCACCGACGCCT

ALS
TCCAGGAGACGCCCATAGTCGAGGTCACCCGCT

Oryza sativa

AGCGGGTGACCTCGACTATGGGCGTCTCCTGGAAGGCGTCGGTG
242

Prol7l Ser
CCGATCATGCGGCGGGAGACCTGGCCCGTGATGGCGACCATCG

CCC-TCC
GGACGGAGTCGAGCAGCGCGTCGGCGAGCGCGGA

GCCAGGTCTCCCGCCGC
243

GCGGCGGGAGACCTGGC
244

Sulfonylurea
CCGCGCTCGCCGACGCGCTGCTCGACTCCGTCCCGATGGTCGCC
245

Resistance
ATCACGGGCCAGGTCCAACGCCGCATGATCGGCACCGACGCCTT

ALS
CCAGGAGACGCCCATAGTCGAGGTCACCCGCTC

Oryza sativa

GAGCGGGTGACCTCGACTATGGGCGTCTCCTGGAAGGCGTCGGT
246

Pro171Gln
GCCGATCATGCGGCGTee TGGACCTGGCCCGTGATGGCGACCATCG

CCC-CAA
GGACGGAGTCGAGCAGCGCGTCGGCGAGCGCGG

CCAGGTCCAACGCCGCA
247

TGCGGCGTTGGACCTGG
248

Sulfonylurea
CCGCGCTCGCCGACGCGCTGCTCGACTCCGTCCCGATGGTCGCC
249

Resistance
ATCACGGGCCAGGTCCAGCGCCGCATGATCGGCACCGACGCCTT

ALS
CCAGGAGACGCCCATAGTCGAGGTCACCCGCTC

Oryza sativa

GAGCGGGTGACCTCGACTATGGGCGTCTCCTGGAAGGCGTCGGT 250

Pro171Gln
GCCGATCATGCGGCGCTGGACCTGGCCCGTGATGGGGACCATCG

CCC-CAG
GGACGGAGTCGAGCAGCGCGTCGGCGAGCGCGG

CCAGGTCCAGCGCCGCA
251

TGCGGCGCTGGACCTGG
252

Imidazolinone
GGCCATACTTGTTGGATATCATCGTCCCGCACCAGGAGCATGTGC
253

Resistance
TGCCTATGATCCCAAATGGGGGCGCATTCAAGGACATGATCCTGG

ALS
ATGGTGATGGCAGGACTGTGTATTAATCTAT

Oryza sativa

ATAGATTAATACACAGTCCTGCCATCACCATCCAGGATCATGTCCT
254

Ser627Asn
TGAATGCGCCCCCATTTGGGATCATAGGCAGCACATGCICCTGGI

AGT-AAT
GCGGGACGATGATATCCAACAAGTATGGCC

GATCCCAAATGGGGGCG
255

CGCCCCGATTTGGGATC
256

Sulfonylurea
TCTGCGCTCGCAGACGCGTTGCTCGACTCCGTCCCCATGGTCGC
257

Resistance
CATCACGGGACAGGTGTCGCGACGCATGATTGGCACCGACGCCT

ALS
TTCAGGAGACGCCCATCGTCGAGGTCACCCGCT

Zea mays

AGCGGGTGACCTCGACGATGGGCGTCTCCTGAAAGGCGTCGGTG
258

Pro165Ser
CCAATCATGCGTCGCGACACCTGTCCCGTGATGGCGACCATGGG

CCG-TCG
GACGGAGTCGAGCAACGCGTCTGCGAGCGCAGA

GACAGGTGTCGCGACGC
259

GCGTCGCGACACCTGTC
260

Sulfonylurea
CTGCGCTCGCAGACGCGTTGCTCGACTCCGTCCCCATGGTCGCC
261

Resistance
ATCACGGGACAGGTGCAGCGACGCATGATTGGCACCGACGCCTT

ALS
TCAGGAGACGCCCATCGTCGAGGTCACCCGCTC

Zea mays

GAGCGGGTGACCTCGACGATGGGCGTCTCCTGAAAGGCGTCGGT 262

Pro165Gln
GCCAATCATGCGTCGCTGCACCTGTCCCGTGATGGCGACCATGG

CCG-CAG
GGACGGAGTCGAGCAACGCGTCTGCGAGCGCAG

ACAGGTGCAGCGACGCA
263

TGCGTCGCTGCACCTGT
264

Imidazolinone
GGCCGTACCTCTTGGATATAATCGTCCCGCACCAGGAGCATGTGT
265

Resistance
TGCCTATGATCCCTAATGGTGGGGCTTTCAAGGATATGATCCTGG

ALS
ATGGTGATGGCAGGACTGTGTATTGATCCGT

Zea mays

ACGGATCAATACACAGTCCTGCCATCACCATCCAGGATCATATCC
266

Ser621Asn
TTGAAAGCCCCACCATTAGGGATCATAGGCAACACATGCTCCTGG

AGT-AAT
TGCGGGACGATTATATCCAAGAGGTACGGCC

GATCCCTAATGGTGGGG
267

CCCCACCATTAGGGATC
268

Sulfonylurea
AGTGGTCTCGCTGATGCAATGCTCGATAGTATCCCTCTCGTGGCG
269

Resistance
ATCACTGGICAAGTCTCTCGTCGGATGATCGGTACCGATGCTTTC

ALS
CAGGAAACTCCAATTGTTGAGGTAACAAGGT

Gossypium hirsutum

ACCTTGTTACCTCAACAATTGGAGTTTCCTGGAAAGCATCGGTAC
270

Pro186Ser
CGATCATCCGACGAGAGACTTGACCAGTGATCGCCACGAGAGGG

CCT-TCT
ATACTATGGAGCATTGCATCAGCGAGACCACT

GTCAAGTCTCTCGTCGG
271

CCGACGAGAGACTTGAC
272

Sulfonylurea
GTGGTCTCGCTGATGCAATGCTCGATAGTATCCCTCTCGTGGCGA
273

Resistance
TCACTGGTCAAGTCCAACGTCGGATGATCGGTACCGATGCTTTCC

ALS
AGGAAACTCCAATTGTTGAGGTAACAAGGTC

Gossypium hirsutum

GACCTTGTTACCTTAACAATTGGAGTTTCCTGGAAAGCATCGGTA
274

Pro186Gln
CCGATCATCCGACGTTGGACTTGACCAGTGATCGCCACGAGAGG

CCT-CAA
GATACTATCGAGCATTGCATCAGCGAGACCAC

TCAAGTCCAACGTCGGA
275

TTCCGACGTTGGACTTGA
276

Sulfonylurea
GTGGTCTCGCTGATGCAATGCTCGATAGTATCCCTCTCGTGCCGA
277

Resistance
TCACTGGTCAAGTCCAGCGTCGGATGATCGGTACCGATGCTTTCC

ALS
AGGAAACTCCAATTGTTGAGGTAACAAGGTC

Gossypium hirsutum

GACCTTGTTACCTCAACAATTGGAGTTTCCTGGAAAGCATCGGTA
278

Pro186Gln
CCGATCATCCGACGCTGGACTTGACCAGTGATCGCCACGAGAGG

CCT-CAG
GATACTATCGAGCATTGCATCAGCGAGACCAC

TCAAGTCCAGCGTCGGA
279

TCCGACGCTGGACTTGA
280

Imidazolinone
GACCTTACTTGTTGGATGTGATTGTCCCACATCAAGAACATGTCCT
281

Resistance
GCCTATGATCCCCAATGGAGGGGCTTTCAAAGATGTGATCACAGA

ALS
GGGTGATGGAAGAACACAATATTGACCTCA

Gossypium hirsutum

TGAGGTCAATATTGTGTTCTTCCATCACCCTCTGTGATCACATCTT
282

Ser642Asn
TGAAAGCCCCTCCATTGGGGATCATAGGCAGGACATGTTCTTGAT

AGT-AAT
GTGGGACAATCACATCCAACAAGTAAGGTC

GATCCCCAATGGAGGGG
283

CCCCTCCATee TGGGGATC
284

Sulfonylurea
TCTGGTCTTGCTGATGCACTTCTTGACTCAGTCCCTCTTGTCGCCA
285

Resistance
TTACTGGGCAAGTTTCCCGGCGTATGATTGGTACTGATGCTTTTCA

ALS
AGAGACTCCAATTGTTGAGGTAACTCGAT

Amaranthus powellii

ATCGAGTTACCTCAACAATTGGAGTCTCTTGAAAAGCATCAGTACC
286

Pro192Ser
AATCATACGCCGGGAAACTTGCCCAGTAATGGCGACAAGAGGGA

CCC-TCC
CTGAGTCAAGAAGTGCATCAGCAAGACCAGA

GGCAAGTTTCCCGGCGT
287

ACGCCGGGAAACTTGCC
288

Sulfonymurea
CTGGTCTTGCTGATGCACTTCTTGACTCAGTCCCTCTTGTCGCCAT
289

Resistance
TACTGGGCAAGTTCAACGGCGTATGATTGGTACTGATGCTTTTCA

ALS
AGAGACTCCAATTGTTGAGGTAACTCGATC

Amaranthus powellii

GATCGAGTTACCTCAACAATTGGAGTCTCTTGAAAAGCATCAGTAC
290

Pro192Gln
CAATCATACGCCGTTGAACTTGCCCAGTAATGGCGACAAGAGGGA

CCC-CAA
CTGAGTCAAGAAGTGCATCAGCAAGACCAG

GCAAGTTCAACGGCGTA
291

TACGCCGTTGAACTTGC
292

Sulfonylurea
CTGGTCTTGCTGATGCACTTCTTGACTCAGTCCCTCTTGTCGCCAT
293

Resistance
TACTGGGCAAGTTCAGCGGCGTATGATTGGTACTGATGCTTTTCA

ALS
AGAGACTCCAATTGTTGAGGTAACTCGATC

Amaranthus powellii

GATCGAGTTACCTCAACAATTGGAGTCTCTTGAAAAGCATCAGTAC
294

Pro192Gln
CAATCATACGCCGCTGAACTTGCCCAGTAATGGCGACAAGAGGG

CCC-CAG
ACTGAGTCAAGAAGTGCATCAGCAAGACCAG

GCAAGTTCAGCGGCGTA
295

TACGCCGCTGAACTTGC
296

Imidazolinone
GACCGTATCTGCTGGATGTAATCGTACCACATCAGGAGCATGTGC
297

Resistance
TGCCTATGATCCCTAACGGTGCCGCCTTCAAGGACACCATAACAG

ALS
AGGGTGATGGAAGAAGGGCTTATTAGTTGGT

Amaranthus powellii

ACCAACTAATAAGCCCTTCTTCCATCACCCTCTGTTATGGIGTCCT
298

Ser652Asn
TGAAGGCGGCACCGTTAGGGATCATAGGCAGCACATGCTCCTGA

AGC-AAC
TGTGGTACGATTACATCCAGCAGATACGGTG

GATCCCTAACGGTGCCG
299

CGGCACCGTTAGGGATC
300

[0121]

14

TABLE 12

Genome-Altering Oligos Conferring Porphyric Herbicide Resistance

Phenotype, Gene,

Plant & Targeted

SEQ ID

Alteration
Altering Oligos
NO:

Porphyric Herbicide
TCTTGCGCCCTCTTTCTGAATCTGCTGCAAATGCACTCTCAAAACT
301

Resistant
ATATTACCCACCAATGGCAGCAGTATCTATCTCGTACCCGAAAGA

PPO
AGCAATCCGAACAGAATGTTTGATAGATGG

Arabidopsis thaliana

CCATCTATCAAACATTCTGTTCGGATTGCTTCTTTCGGGTACGAGA
302

Val365Met
TAGATACTGCTGCCATTGGTGGGTAATATAGTTTTGAGAGTGCATT

GTT-ATG
TGCAGCAGATTCAGAAAGAGGGCGCAAGA

CCCACCAATGGCAGCAG
303

CTGCTGCCATTGGTGGG
304

Porphyric Herbicide
TATTACGTCCTCTTTCGGTTGCCGCAGCAGATGCACTTTCAAATTT
305

Resistant
CTACTAICCCCCAATGGGAGCAGTCACAATTTCATATCCTCAAGAA

PPO
GCTATTCGTGATGAGCGTCTGGTTGATGG

Nicotiana tabacum

CCATCAACCAGACGCTCATCACGAATAGCTTCTTGAGGATATGAA
306

Val376Met
ATTGTGACTGCTCCCATTGGGGGATAGTAGAAATTTGAAAGTGCA

GTT-ATG
TCTGCTGCGGCAACCGAAAGAGGACGTAATA

TCCCCCAATGGGAGCAG
307

CTGCTCCCATTGGGGGA
308

Porphyric Herbicide
TGTTGCGTCCGCTTTCGTTGGGTGCAGCAGATGCATTGTCAAAAT
309

Resistant
TTTATTATCCTCCGATGGCAGCTGTATCAATTTCATATCCAAAAGA

PPO
CGGAATTCGTGCTGACCGGCTGATTGATGG

Cichorium intybus

CCATCAATCAGCCGGTCAGCACGAATTGCGTCTTTTGGATATGAA
310

Val383Met
ATTGATACAGCTGCCATCGGAGGATAATAAAATTTTGACAATGCAT

GTT-ATG
CTGCTGCACCCAACGAAAGCGGACGCAACA

TCCTCCGATGGCAGCTG
311

CAGCTGCCATCGGAGGA
312

Porphyric Herbicide
TCCTTCGTCCACTTTCAGATGTCGCCGCAGAATCTCTTTCAAAATT
313

Resistant
TCATTATCCACCAATGGCAGCTGTGTCACTTTCCTATCCTAAAGAA

PPO
GCAATTAGATCAGAGTGCTTGATTGACGG

Spinacia oleracea

CCGTCAATCAAGCACTCTGATCTAATTGCTTCTTTAGGATAGGAAA
314

Val390Met
GTGACACAGCTGCCATTGGTGGATAATGAAATTTTGAAAGAGATT

GTT-ATG
CTGCGGCGACATCTGAAAGTGGACGAAGGA

TCCACCAATGGCAGCTG
315

CAGCTGCCATTGGTGGA
316

Porphyric Herbicide
TTTTGCGTCCACTTTCAAGCGATGCTGCAGATGCTCTATCAAGATT
317

Resistant
CTATTATCCACCGATGGCTGCIGTAACTGTTTCGTATCCAAAGGAA

PPO
GCAATTAGAAAAGAATGCTTAATTGATGG

Zea mays

CGATCAATTAAGCATTCTTTTCTAATTGCTTCCTTTGGATACGAAAC
318

Val363Met
AGTTACAGCAGCCATCGGTGGATAATAGAATCTTGATAGAGCATC

GTT-ATG
TGCAGCATCGCTTGAAAGTGGACGCAAAA

TCCACCGATGGCTGCTG
319

CAGCAGCCATCGGTGGA
320

Porphyric Herbicide
TCTTGCGGCCACTTTCAAGTGATGGAGCAGATGCTCTGTCAATATT
321

Resistant
CTATTATCCACCAATGGCTGCTGTAACTGTTTCATATCCAAAAGAA

PPO
GCAATTAGAAAAGAATGCTTAATTGACGG

Oryza sativa

CCGTCAATTAAGCATTCTTTTCTAATTGCTTCTTTTGGATATGAAAC
322

Val364Met
AGTTACAGCAGCCATTGGTGGATAATAGAATATTGACAGAGCATC

GTT-ATG
TGCTGCATCACTTGAAAGTGGCCGCAAGA

TCCACCAATGGCTGCTG
323

CAGCAGCCATTGGTGGA
324

Porphyric Herbicide
CTGGTCAAGGAGCAGGCGCCCGCCGCCGCCGAGGCCCTGGGCT
325

Resistant
CCTTCGACTACCCGCCGATGGGCGCCGTGACGCTGTCGTACCCG

PPO
CTGAGCGCCGTGCGGGAGGAGCGCAAGGCCTCGG

Chlamydomonas

CCGAGGCCTTGCGCTCCTCCCGCACGGCGCTCAGCGGGTACGAC
326

reinhardtii

AGCGTCACGGCGCCCATCGGCGGGTAGTCGAAGGAGCCCAGGG

Val389Met
CCTCGGCGGCGGCGGGCGCCTGCTCCTTGACCAG

GTG-ATG
ACCCGCCGATGGGCGCC
327

GGCGCCCATCGGGGGGT
328

[0122]

15

TABLE 13

Genome-Altering Oligos Conferring Triazine Resistance

Phenotype, Gene,

Plant & Targeted

SEQ ID

Alteration
Altering Oligos
NO:

Triazine Resistant
AAACTTACAACATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT
329

D1 Protein
TTTCCAATATGCTACTTTCAACAATTCTCGTTCTTTACATTTCTTCTT

Arabidopsis thaliana

AGCGGCTTGGCCGGTAGTAGGTATTTG

Ser264Thr
CAAATACCTACTACCGGCCAAGCCGCTAAGAAGAAATGTAAAGAA
330

AGT-ACT
CGAGAATIGTTGAAAGTAGCATATTGGAAAATCAATCGGCCAAAAT

AACCGTGAGCAGCTACAATGTTGTAAGTTT

ATATGCTACTTTCAACA
331

TGTTGAAAGTAGCATAT
332

Triazine Resistant
AAACTTATAACATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT
333

D1Protein
CTTCCAATATGCTACTTTCAACAACTCTCGTTCGTTACACTTCTTCC

Nicotiana tabacum

TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr
CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAGTGTAACGAA
334

AGT-ACT
CGAGAGtTGTIGAAAGTAGCATATTGGAAGATCAAtCGGCCAAAA

TAACCATGAGCGGCTACGATGTTATAAGTTT

ATATGCTACTTTCAACA
335

TGTTGAAAGTAGCATAT
336

Triazine Resistant
AAACTTATAATATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT
337

D1Protein
CTTCCAATATGCTACTTTTAACAACTCTCGCTCTTTACATTTCTTCT

Populus deltoides

TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr
CAGATACCTACTACAGGCCAAGCAGCTAAGAAGAAATGTAAAGAG
338

AGT-ACT
CGAGAGTTGTTAAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCGGCTACGATATTATAAGTTT

ATATGCTACTTTTAACA
339

TGTTAAAAGTAGCATAT
340

Triazine Resistant
AAACTTATAATATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT
341

D1Protein
CTTCCAATATGCTACTTTCAACAACTCTCGTTCGTTACACTTCTTCC

Petunia x hybrida

TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr
CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAGTGTAACGAA
342

AGT-ACT
CGAGAGTTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCGGCTACGATATTATAAGTTT

ATATGCTACTTTCAACA
343

TGTTGAAAGTAGCATAT
344

Triazine Resistant
AAACTTATAAIATCGTAGCTGCTCATGGTTATTTTGGCCGATTGAT
345

D1Protein
CTTCCAATATGCTACTTTCAACAATTCTCGTTCTTTACATTTCTTCC

Magnolia pyramidata

TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr
CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAATGTAAAGAA
346

AGT-ACT
CGAGAATTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCAGCTACGATATTATAAGTTT

ATATGCTACTTTCAACA
347

TGTTGAAAGTAGCATAT
348

Triazine Resistant
AAACCTATAATATTGTAGCAGCTCATGGTTATTTTGGCCGATTGAT
349

D1Protein
CTTCCAATATGCTACTTTCAACAACTCTCGTTCTTTACATTTCTTCC

Medicago sativa

TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr
CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAATGTAAAGAA
350

AGT-ACT
CGAGAGTTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAAGCATGAGCTGCTACAATATTATAGGTTT

ATATGCTACTTTCAACA
351

TGTTGAAA+E,us GTAGCATAT
1352

Triazine Resistant
AAACCTATAATATTGTAGCTGCTCATGGTTATTTGGCCGATTGAT
353

D1Protein
CTTCCAATATGCAACTTTCAACAATTCTCGTTCTTTACATTTCTTCT

Glycine max

TAGCTGCTTGGCCTGTAGTAGGTATTTG

Ser264Thr
CAAATACCTACTACAGGCCAAGCAGCTAAGAAGAAATGTAAAGAA
354

AGT-ACT
CGAGAATTGTTGAAAGTTGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCAGCTACAATATTATAGGTTT

ATATGCAACTTTCAACA
355

TGTTGAAAGTTGCATAT
356

Triazine Resistant
AAACTTACAACATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT
357

D1Protein
CTTCCAATATGCTACTTTCAACAATTCTCGTTCTTTACATTTCTTCT

Brassica napus

TAGCGGCTTGGCCGGTAGTAGGTATTTG

Gly264Thr
CAAATACCTACTACCGGCCAAGCCGCTAAGAAGAAATGTAAAGAA
358

GGT-ACT
CGAGAAITGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCGTGAGCAGCTACAATGTTGTAAGTTT

ATATGCTACTTTCAACA
359

TGTTGAAAGTAGCATAT
360

Triazine Resistant
AAACTTATAATATTGTGGCCGCTCATGGTTATTTTGGCCGATTAAT
361

D1Protein
CTTCCAATATGCTACTTTTAACAACTCTCGTTCTTTACACTTCTTCT

Oryza sativa

TGGCTGCTTGGCCTGTAGTAGGGATTTG

Ser264Thr
CAAATCCCTACTACAGGCCAAGCAGCCAAGAAGAAGTGTAAAGAA
362

AGT-ACT
CGAGAGTTGTTAAAAGTAGCATATTGGAAGATTAATCGGCCAAAAT

AACCATGAGCGGCCACAATATTATAAGTTT

ATATGCTACTTTTAACA
363

TGTTAAAAGTAGCATAT
364

Triazine Resistant
AGACTTATAATATTGTGGCTGCTCACGGTTATTTTGGTCGATTAAT
365

D1Protein
CTTCCAATATGCTACTTTCAACAATTCTCGTTCTTTACACTTCTTCT

Zea mays

TGGCTGCTtGGCCTGTAGTAGGGATCtG

Ser264Thr
CAGATCCCTACTACAGGCCAAGCAGCCAAGAAGAAGTGTAAAGAA
366

AGT-ACT
CGAGAATTGTTGAAAGTAGCATATTGGAAGATTAATCGACCAAAAT

AACCGTGAGCAGCCACAATATTATAAGTCT

ATATGCTACTTTCAACA
367

TGTTGAAAGTAGCATAT
368

Triazine Resistant
AAACTTACAACATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT
369

D1Protein
TTTCCAATATGCTACTTTCAACAATTCTCGTTCTTTACATTTCTTCTT

Arabidopsis thaliana

AGCGGCTTGGCCGGTAGTAGGTATTTG

Ser264Thr
CAAATACCTACTACCGGCCAAGCCGCTAAGAAGAAATGTAAAGAA
370

AGT-ACT
CGAGAATTGITGAAAGTAGCATATTGGAAAATCAATCGGCCAAAAT

AACCGTGAGCAGCTACAATGTTGTAAGTTT

ATATGCTACTTTCAACA
371

TGTTGAAAGTAGCATAT
372

Triazine Resistant
AAACTTATAACATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT
373

D1Protein
CTTCCAATATGCTACTTTCAACAACTCTCGTTCGTTACACTTCTTCC

Nicotiana tabacum

TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr
CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAGTGTAACGAA
374

AGT-ACT
CGAGAGTTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCGGCTACGATGTTATAAGTTT

ATATGCTACTTTCAACA
375

TGTTGAAAGTAGCATAT
376

Triazine Resistant
AAACTTATAATATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT
377

D1Protein
CTTCCAATATGCTACTTTTAACAACTCTCGCTCTTTACATTTCTTCT

Papulus deltoides

TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr
CAGATACCTACTACAGGCCAAGCAGGTAAGAAGAAATGTAAAGAG
378

AGT-AGT
CGAGAGTTGTTAAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCGGCTACGATATTATAAGTTT

ATATGCTACTTTTAACA
379

TGTTAAAAGTAGCATAT
380

Triazine Resistant
AAACTTATAATATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT
381

D1Protein
CTTCCAATATGCTACTTTCAACAACTCTCGTTCGTTACACTTCTTCC

Petunia x hybrida

TAGCTGGTTGGCCTGTAGTAGGTATCTG

Ser264Thr
CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAGTGTAACGAA
382

AGT-ACT
CGAGAGTTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCGGCTACGATATTATAAGTTT

ATATGCTACTTTCAACA
383

TGTTGAAAGTAGCATAT
384

Triazine Resistant
AAACTTATAATATCGTAGCTGCTCATGGTTATTTTGGCCGATTGAT
385

D1Protein
CTTCCAATATGCTACTTTCAACAATTCTCGTTCTTTACATTTCTTCC

Magnolia pyramidata

TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr
CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAATGTAAAGAA
386

AGT-ACT
CGAGAATTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCAGCTACGATATTATAAGTTT

ATATGCTACTTTCAACA
387

TGTTGAAAGTAGCATAT
388

Triazine Resistant
AAACCTATAATATTGTAGCAGCTCATGGTTATTTTGGCCGATTGAT
389

D1Protein
CTTCCAATATGCTACTTTCAACAACTCTCGTTCTTTACATTTGTTCC

Medicago sativa

TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr
CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAATGTAAAGAA
390

AGT-ACT
CGAGAGTTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCTGCTACAATATTATAGGTTT

ATATGCTACTTTCAACA
391

TGTTGAAAGTAGCATAT
392

Triazine Resistant
AAACCTATAATATTGTAGCTGCTCATGGTTATTTTGGCCGATTGAT
393

D1Protein
CTTCCAATATGCAACTTTCAACAATTCTCGTTCTTTACATTTCTTCT

Glycine max

TAGCTGCTTGGCCTGTAGTAGGTATTTG

Ser264Thr
CAAATACCTACTACAGGCCAAGCAGCTAAGAAGAAATGTAAAGAA
394

AGT-ACT
CGAGAATTGTTGAAAGTTGCATATTGGAAGATCAATCGGGCAAAA

TAACCATGAGCAGCTACAATATTATAGGTTT

ATATGCAACTTTCAACA
395

TGTTGAAAGTTGCATAT
396

Triazine Resistant
AAACTTACAACATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT
397

D1Protein
CTTCCAATATGCTACTTTCAACAATTCTCGTTCTTTACATTTCTTCT

Brassica napus

TAGCGGCTTGGCCGGTAGTAGGTATTTG

Gly264Thr
CAAATACCTACTACCGGCCAAGCCGCTAAGAAGAAATGTAAAGAA
398

GGT-ACT
CGAGAATTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCGTGAGCAGCTACAATGTTGTAAGTTT

ATATGCTACTTTCAACA
399

TGTTGAAAGTAGCATAT
400

Triazine Resistant
AAACTTATAATATTGTGGCCGCTCATGGTTATTTTGGCCGATTAAT
401

D1Protein
CTTCCAATATGCTACTTTTAACAACTCTCGTTCTTTACACTTCTTCT

Oryza sativa

TGGCTGCTTGGCCTGTAGTAGGGATTTG

Ser264Ihr
CAAATCCCTACTACAGGCCAAGCAGCCAAGAAGAAGTGTAAAGAA
402

AGT-ACT
CGAGAGTTGTTAAAAGTAGCATATTGGAAGATTAATCGGCCAAAAT

AACCATGAGCGGCCACAATATTATAAGTTT

ATATGCTACTTTTAACA
403

TGTTAAAAGTAGCATAT
404

Triazine Resistant
AGACTTATAATATTGTGGCTGCTCACGGTTATTTTGGTCGATTAAT
405

D1Protein
CTTCCAATATGCTACTTTCAACAATTCTCGTTCTTTACACTTCTTCT

Zea mays

TGGCTGCTTGGCCTGTAGTAGGGATCTG

Ser264Thr
CAGATCCCTACTACAGGCCAAGCAGCCAAGAAGAAGTGTAAAGAA
406

AGT-ACT
CGAGAATTGTTGAAAGTAGCATATTGGAAGATTAATCGACCAAAAT

AACCGTGAGCAGCCACAATATTATAAGTCT

ATATGCTACTTTCAACA
407

TGTTGAAAGTAGCATAT
408

Triazine Resistant
AAACTTACAACATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT
409

D1Protein
TTTCCAATATGCTACTTTCAACAATTCTCGTTCTTTACATTTCTTCTT

Arabidopsis thaliana

AGCGGCTTGGCCGGTAGTAGGTATTTG

Ser264Thr
CAAATACCTACTACCGGCCAAGCCGCTAAGAAGAAATGTAAAGAA
410

AGT-ACT
CGAGAATTGTTGAAAGTAGCATATTGGAAAATCAATCGGCCAAAAT

AACCGTGAGCAGCTACAATGTTGTAAGTTT

ATATGCTACTTTCAACA
411

TGTTGAAAGTAGCATAT
412

Triazine Resistant
AAACCTACAATATTGTGGCTGCTCACGGTTATTTCGGCCGATTGAT
413

D1Protein
CTTCCAGTATGCTACTTTCAACAACTCCCGTTCTTTACATTTCTTCT

Picea abies

TAGCTGCTTGGCCCGTAGCAGGTATCTG

Ser264Thr
CAGATACCTGCTACGGGCCAAGCAGCTAAGAAGAAATGTAAAGAA
414

AGT-ACT
CGGGAGTTGTTGAAAGTAGCATACTGGAAGATCAATCGGCCGAAA

TAACCGTGAGCAGCCACAATATTGTAGGTTT

GTATGCTACTTTCAACA
415

TGTTGAAAGTAGCATAC
416

Triazine Resistant
AAACCTATAATATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT
417

D1Protein
CTTCCAATATGCTACTTTCAACAATTCTCGCTCTTTACATTTCTTCC

Vicia faba

TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr
CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAATGTAAAGAG
418

AGT-ACT
CGAGAATTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCGTGAGCAGCTACAATATTATAGGTTT

ATATGCTACTTTCAACA
419

TGTTGAAAGTAGCATAT
420

Triazine Resistant
AGACTTATAATATTGTGGCTGCTCATGGTTATTTTGGCCGATTAAT
421

D1Protein
CTTCCAATATGCTACTTTCAACAACTCTCGTTCTTTACACTTCTTCT

Hordeum vulgare

TGGCTGCTTGGCCTGTAGTAGGAATCTG

Ser264Thr
CAGATTCCTACTACAGGCCAAGCAGCCAAGAAGAAGTGTAAAGAA
422

AGT-ACT
CGAGAGTTGTTGAAAGTAGCATATTGGAAGATTAATCGGCCAAAA

TAACCATGAGCAGCCACAATATTATAAGTCT

ATATGCTACTTTCAACA
423

TGTTGAAAGTAGCATAT
424

Triazine Resistant
AAACTTATAATATTGTGGCTGCTCATGGTTATTTTGGCCGATTAAT
425

D1Protein
CTTCCAATATGCTACTTTCAACAACTCTCGTTCTTTACACTTCTTCT

Triticum aestivum

TGGCTGCTTGGCCTGTAGTAGGAATCTG

Ser264Thr
CAGATTCCTACTACAGGCCMGCAGCCAAGAAGAAGTGTAAAGAA
426

AGT-ACT
CGAGAGTTGTTGAAAGTAGCATATTGGAAGATTAATCGGCCAAAA

TAACCATGAGCAGCCACAATATTATAAGTTT

ATATGCTACTTTCAACA
427

TGTTGAAAG+E TAGCATAT
428

Triazine Resistant
AAACTTATAATATTGTAGCTGCTCATGGTTATTTTGGCCGATTAATC
429

D1Protein
TTCCAATATGCAACTTTCMCAATTCTCGTTCTTTACATTTCTTCCT

Vigna unguiculata

AGCTGCTTGGCCTGTAGTAGGTATTTG

Ser264Thr
CAAATACCTACTACAGGCCAAGCAGCTAGGAAGAAATGTAAAGAA
430

AGT-ACT
CGAGAATTGTTGAAAGTTGCATATTGGAAGATTAATCGGCCAAAAT

AACCATGAGCAGCTACAATATTATAAGTTT

ATATGCAACTTTCAACA
431

TGTTGAAAGTTGCATAT
432

Triazine Resistant
AAACCTATAATATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT
433

D1Protein
CTTCCAATATGCAACTTTCAACAACTCTCGTTCTTTACACTTCTTCT

Lotus japonicus

TAGCTGCTTGGCCTGTTGTAGGTATCTG

Ser264Thr
CAGATACCTACAACAGGCCAAGCAGCTAAGAAGAAGTGTAAAGAA
434

AGT-ACT
CGAGAGTTGTTGAAAGTTGCATATTGGAAGATCAATCGGCCAAAA

TAACCGTGAGCAGCTACAATATTATAGGTTT

ATATGCAACTTTCAACA
435

TGTTGAAAGTTGCATAT
436

Triazine Resistant
AAACTTACAACATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT
437

D1Protein
CTTCCAATATGCTACTTTCAACAATTCTCGTTCTTTACATTTCTTCT

Sinapis alba

TAGCGGCTTGGCCGGTAGTAGGTATTTG

Ser264Thr
CAAATACCTACTACCGGCCAAGCCGCTAAGAAGAAATGTAAAGAA
438

AGT-ACT
CGAGAATTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCGTGAGCAGCTACAATGTTGTAAGTTT

ATATGCTACTTTCAACA
439

TGTTGAAAGTAGCATAT
440

Triazine Resistant
AAACCTATAATATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT
441

D1Protein
CTTCCAATATGCTACTTTCAACAATTCTCGCTCTTTACATTTCTTCC

Pisum sativum

TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr
CAGATACCTACTACAGGCCAAGCAGCTIAGGAAGAAATGTAAAGAG
442

AGt-ACT
CGAGAATTGTTGAAAGTAGCAtATTGGAAGATCAATCGGCCAAAA

TAACCGTGAGCAGCTACAATATTATAGGTTT

ATATGCTACTTTCAACA
443

TGTTGAAAGTAGCATAT
444

Triazine Resistant
AAACTTATAATATCGTAGGTGCTCATGGTTATTTTGGTCGATTGAT
445

D1Protein
CTTCCAATATGCTACTTTCAACAACTCTCGTTCTTTACACTTCTTCT

Spinacia oleracea

TAGCTGCTTGGCCTGIAGTAGGTATTTG

Ser264Thr
CAAATACCTACTACAGGCCAAGCAGCTAAGAAGAAGTGTAAAGAA
446

AGT-ACT
CGAGAGTTGTTGAAAGTAGCATATTGGAAGATCAATCGACCAAAA

TAACCATGAGCAGGTACGATATTATAAGTTT

ATATGCTACTTTCAACA
447

TGTTGAAAGTAGCATAT
448

Triazine Resistant
AAACTTATAACATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT
449

D1Protein
CTTCCAATATGCTACTTTCAACAACTCTCGTTCGTTACACTTCTTCC

Nicotiana debneyi

TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr
CAGAtACCtACTACAGGCGAAGCAGCtAGGAAGAAGTGTAACGAA
450

AGT-ACT
CGAGAGtTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCGGCTACGATGTTATAAGTTT

ATATGCTACTTTCAACA
451

TGTTGAAAGTAGCATAT
452

Triazine Resistant
AAACTTATAATATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT
453

D1Protein
CTTCCAATATGCTACTTTCAACAACTCTCGTTCGTTACACTTCTTCC

Solanum nigrum

TAGCTGCTTGGCCTGTAGTAGGTATCTG

Ser264Thr
CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAGTGTAACGAA
454

AGT-ACT
CGAGAGTTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

TAACCATGAGCGGCTACGATATTATAAGTTT

ATATGCTACTTTCAACA
455

TGTTGAAAGTAGCATAT
456

Triazine Resistant
AAACTTATAACATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT
457

D1Protein
CTTCCAATATGCTACTTTCAACAACTCTCGTICGTTACACTTCTTCC

Nicotiana

TAGCTGCTTGGCCTGTAGTAGGTATCTG

plumbaginifolia

CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAGTGTAACGAA
458

Ser264Thr
CGAGAGTTGTTGAAAGTAGCATATTGGAAGATCAATCGGCCAAAA

AGT-ACT
TAACCATGAGCGGCTACGATGTTATAAGTTT

ATATGCTACTTTCAACA
459

TGTTGAAAGTAGCATAT
460

EXAMPLE 6

Engineering Male- or Female-Sterile Plants

[0123] Flower development in distantly related dicot plant species is increasingly better understood and appears to be regulated by a family of genes which encode regulatory proteins. These genes include, for example, AGAMOUS (AG), APETALA1 (AP1), and APETALA3 (AP3) and PISTILLATA (PI) in Arabidopsis thaliana, and DEFICIENS A (DEFA), GLOBOSA (GLO), SQUAMOSA (SQUA), and PLENA (PLE) in Antirrhinum majus. Genetic studies have shown that the DEFA, GLO and AP3 genes are essential for petal and stamen development. Sequence analysis of these genes revealed that the gene products contain a conserved MADS box region, a DNA-binding domain. Using these clones as probes, MADS box genes have also been isolated from other species including tomato, tobacco, petunia, Brassica napus, and maize.

[0124] Altering the expression of these genes results in altered floral morphology. For example, mutations in AP3 and PI result in male-sterile flowers because petals develop in place of stamens.

[0125] The attached tables disclose exemplary oligonucleotide base sequences which can be used to generate site-specific mutations that confer altered floral structures in plants.

16TABLE 14Oligonucleotides to produce male-sterile plantsPhenotype, Gene,Plant & TargetedSEQ IDAlterationAltering OligosNO:Male-sterileTTGTCCTCTCCACCAAATCTCTTCAACAAAAAGATTAAACAAAGAG461AP3AGAAGAATATGGCGTGAGGGAAGATCCAGATCAAGAGGATAGAGAArabidopsis thalianaACCAGACAAACAGACAAGTGACGTATTCAAArg3TermTTGAATACGTCACTTGTCTGTTTGTCTGGTTCTCTATCCTCTTGATC462AGA-TGATGGATCTTCCCTCACGCCATATTCTTCTCTCTTIGTTTAATCTTTTTGTTGAAGAGATTTGGTGGAGAGGACAAATATGGCGTGAGGGAAG463CTTCCCTCACGCCATAT464Male-sterileTCTCCACCAAATCTCTTCAACAAAAAGATTAAACAAAGAGAGAAGA465AP3ATATGGCGAGAGGGTAGATCCAGATCAAGAGGATAGAGAACCAGAArabidopsis thalianaCAAACAGACAAGTGACGTATTCAAAGAGAALys5TermTTCTCTTTGAATACGTCACTTGTCTGTTTGTCTGGTTCTCTATCCTC466AAG-TAGTTGATCTGGATCTACCCTCTCGCCATATTCTTCTCTCTTTGTTTAATCTTTTTGTTGAAGAGATTTGGTGGAGACGAGAGGGTAGATCCAG467CTGGATCTACCGTCTCG468Male-sterileCCAAATCTCTTCAACAAAAAGATTAAACAAAGAGAGAAGAATATGG469AP3CGAGAGGGAAGATCTAGATCAAGAGGATAGAGAAGCAGACAAACAArabidopsis thalianaGACAAGTGACGTATTCAAAGAGAAGGAATGGln7TermCATTCCTTCTCTTTGAATACGTCACTTGTCTGTTTGTCTGGTTCTCT470CAG-TAGATCCTCTTGATCTAGATCTTCCCTCTCGCCATATTCTTCTCTCTTTGTTTAATCTTTTTGTTGAAGAGATTTGGGGAAGATCTAGATCAAG471CTTGATCTAGATCTTCC472Male-sterileCTCTTCAACAAAAAGATTAAACAAAGAGAGAAGAATATGGCGAGAG473AP3GGAAGATCCAGATCTAGAGGATAGAGAACCAGACAAACAGAGAAGArabidopsis thalianaTGACGTATTCAAAGAGAAGGAATGGTTTATLys9TermATAAACCATTCGTTCTCTTTGAATACGTCACTTGTCTGTTTGTCTGG474AAG-TAGTTCTCTATCCTCTAGATCTGGATCTTCCCTCTCGCCATATTCTTCTCTCTTTGTTTAATCTTTTTGTTGAAGAGTCCAGATCTAGAGGATA475TATCCTCTAGATCTGGA476Male-sterileAGAGGGAAGATCGAGATGAAGAGGATAGAGAACGAGAGGAACCG477AP3ACAAGTGACGTATTCTTAGAGAAGAAATGGTTTGTTCAAGAAAGCTBrassica oleraceaCACGAGCTTACAGTTTTATGTGATGCTAGGGLys23TermCCCTAGCATCACATAAAACTGTAAGCTCGTGAGCTTTCTTGAACAA478AAG-TAGACCATTTCTTCTCTAAGAATACGTCACTTGTCGGTTGGTCTGGTTCTCTATCCTCTTGATCTGGATCTTCCCTCTCGTATTCTTAGAGAAGA479TCTTCTCTAAGAATACG480Male-sterileGGGAAGATCCAGATCAAGAGGATAGAGAACCAGACCAACCGACAA481AP3GTGACGTATTCTAAGTGAAGAAATGGTTTGTTCAAGAAAGCTCACGBrassica oleraceaAGCTTACAGTTTTATGTGATGCTAGGGTTTArg24TermAAACCCTAGCATCACATAAAACTGTAAGCTCGTGAGCTTTCTTGAA482AGA-TGACAAACCATTTCTTCACTTAGAATACGTCACTTGTGGGTTGGTCTGGTTCTCTATCCTCTTGATCTGGATCTTCCCATTCTAAGTGAAGAAAT483ATTTCTTCACTTAGAAT484Male-sterileAAGATCCAGATCAAGAGGATAGAGAACCAGACCAACCGACAAGTG485AP3ACGTATTCTAAGAGATGAAATGGTTTGTTCAAGAAAGCTCACGAGCBrassica oleraceaTTACAGTTTTATGTGATGCTAGGGTTTCGAArg25TermTCGAAACCCTAGCATCACATAAAACTGTAAGCTCGTGAGCTTTCTT486AGA-TGAGAACAAACCATTTCATCTCTTAGAATACGTCACTTGTCGGTTGGTCTGGTTCTCTATGCTCTTGATCTGGATCTTCTAAGAGATGAAATGGT487ACCATTTCATCTCTTAG488Male-sterileTCAAGAGGATAGAGAACCAGACCAACCGACAAGTGACGTATTCTA489AP3AGAGAAGAAATGGTTAGTTCAAGAAAGCTCACGAGCTTACAGTTTTBrassica oleraceaATGTGATGCTAGGGTTTCGATTATCATGTTLeu28TermAACATGATAATCGAAACCCTAGCATCACATAAAACTGTAAGCTCGT490TTG-TAGGAGCTTTCTTGAACTAACCATTTCTTCTCTTAGAATACGTCACTTGTCGGTTGGTCTGGTTCTCTATCCTCTTGAAAATGGTTAGTTCAAGA491TCTTGAACTAACCATTT492Male-sterileGGCTCGAGGGAAGATCCAGATTAAGAGGATAGAGAACCAAACAAA493AP3CAGGCAGGTCACCTAGTCCAAGAGAAGAAATGGTTTGTTCAAGAABrassica napusAGCACACGAGCTCTCTGTTCTCTGTGATGCTTyr21TermAGCATCACAGAGAACAGAGAGCTCGTGTGCTTTCTTGAACAAACC494TAC-TAGATTTCTTCTCTTGGACTAGGTGACCTGCCTGTTTGTTTGGTTCTCTATCCTCTTAATCTGGATCTTCCCTCGAGCCGTCACCTAGTCCAAGAG495CTCTTGGACTAGGTGAC496Male-sterileCGAGGGAAGATCCAGATTAAGAGGATAGAGAACCAAACAAACAGG497AP3CAGGTCACCTACTCCTAGAGAAGAAATGGTTTGTTCAAGAAAGCACBrassica napusACGAGCTCTCTGTTCTCTGTGATGCTAAAGLys23TermCTTTAGCATCACAGAGAACAGAGAGCTCGTGTGCTTTCTTGAACAA498AAG-TAGACCATTTCTTCTCTAGGAGTAGGTGACCTGCCTGTTTGTTTGGTTCTCTATCCTCTTAATCTGGATCTTCCCTCGCCTACTCCTAGAGAAGA499TCTTCTCTAGGAGTAGG500Male-sterileGGGAAGATCCAGATTAAGAGGATAGAGAACCAAACAAACAGGCAG501AP3GTCACCTACTCCAAGTGAAGAAATGGTTTGTTCAAGAAAGCACACGBrassica napusAGCTCTCTGTTCTCTGTGATGCTAAAGTTTArg24TermAAACTTTAGCATCACAGAGAACAGAGAGCTCGTGTGCTTTCTTGAA502AGA-TGACAAACCATTTGTTCACTTGGAGTAGGTGACCTGCCTGTTTGTTTGGTTCTCTATCCTCTTAATCTGGATCTTCCCACTCCAAGTGAAGAAAT503ATTTCTTCACTTGGAGT504Male-sterileAAGATCCAGATTAAGAGGATAGAGAACCAAACAAACAGGCAGGTC505AP3ACCTACTCCAAGAGATGAAATGGTTTGTTCAAGAAAGCACACGAGBrassica napusCTCTCTGTTCTCTGTGATGCTAAAGTTTCCAArg25TermTGGAAACTTTAGCATCACAGAGAACAGAGAGCTCGTGTGCTTTCTT506AGA-TGAGAACAAACCATTTCATCTCTTGGAGTAGGTGACCTGCCTGTTTGTTTGGTTCTCTATCCTCTTAATCTGGATCTTCCAAGAGATGAAATGGT507ACCATTTCATCTCTTGG508Male-sterileGGAGAGAAAGGAAAGCTGGAAGAAGAAAACAAGAGCAGTAGTGG509DEFATAGTGGTTCGATGGCTTGAGGGAAGATCCAGATTAAGAGGATAGAAntirrhinum majusGAACCAAACAAACAGGCAGGTCACCTACTCCAArg3TermTGGAGTAGGTGACCTGCCTGTTTGTTTGGTTCTCTATCCTCTTAAT510CGA-TGACTGGATCTTCCCTCAAGCCATCGAACCACTACCACTACTGCTCTTGTTTTCTTCTTCCAGCTTTCCTTTCTCTCCCGATGGCTTGAGGGAAG511CTTCCCTCAAGCCATCG512Male-sterileAAAGGAAAGCTGGAAGAAGAAAACAAGAGCAGTAGTGGTAGTGGT513DEFATCCATGGCTCGAGGGTAGATCCAGATTAAGAGGATAGAGAACCAAAntirrhinum majusACAAACAGGCAGGTCACCTACTCCAAGAGAALys5TermTTCTCTTGGAGTAGGTGACCTGCCTGTTTGTTTGGTTCTCTATCCT514AAG-TAGCTTAATCTGGATCTACCCTCGAGCCATCGAACCACTAGCACTACTGCTCTTGTTTTCTTCTTCCAGCTTTCCTTTCTCGAGGGTAGATCCAG515CTGGATCTACCCTCGAG516Male-sterileAAGCTGGAAGAAGAAAACAAGAGCAGTAGTGGTAGTGGTTCGATG517DEFAGCTCGAGGGAAGATCTAGATTAAGAGGATAGAGAACCAAACAAACAntirrhinum majusAGGCAGGTCACCTACTCCAAGAGAAGAAATGGln7TermCATTTCTTCTCTTGGAGTAGGTGACCTGCCTGTTTGTTTGGTTCTC518CAG-TAGTATCCTCTTAATCTAGATCTTCCCTCGAGCCATCGAACCACTACCACTACTGCTCTTGTTTTCTTCTTCCAGCTTGGAAGATCTAGATTAAG519CTTAATCTAGATCTTCC520Male-sterileGAAGAAGAAAACAAGAGCAGTAGTGGTAGTGGTTCGATGGCTCGA521DEFAGGGAAGATCCAGATTTAGAGGATAGAGAACCAAACAAACAGGCAGAntirrhinum majusGTCACCTACTCCAAGAGAAGAAATGGTTTGTLys9TermACAAACCATTTCTTCTCTTGGAGTAGGTGACCTGCCTGTTTGTTTG522AAG-TAGGTTCTCTATCCTCTAAATCTGGATCTTCCCTCGAGCCATCGAACCACTACCACTACTGCTCTTGTTTTCTTCTTCTCCAGATTTAGAGGATA523TATCCTCTAAATCTGGA524Male-sterileTCAGTAATTCTTAAGATCTCAAACTTTGAGCAAAAAGAAAAAAAAAC525AP3TATGGCTCGTGGGTAGATCCAGATCAAGAGAATAGAGAACCAAACNicotiana tabacumAAACAGACAAGTCACTTATTCTAAGAGAALys5TermTTCTCTTAGAATAAGTGACTTGTCTGTTTGTTTGGTTCTCTATTCTC526AAG-TAGTTGATCTGGATCTACCCACGAGCCATAGTTTTTTTTTCTTTTTGCTCAAAGTTTGAGATCTTAAGAATTACTGACTCGTGGGTAGATCCAG527CTGGATCTACCCACGAG528Male-sterileATTCTTAAGATCTCAAACTTTGAGCAAAAAGAAAAAAAAACTATGGC529AP3TCGTGGGAAGATCTAGATCAAGAGAATAGAGAACCAAACAAACAGNicotiana tabacumACAAGTCACTTATTCTAAGAGAAGAAATGGln7TermCATTTCTTCTCTTAGAATAAGTGACTTGTCTGTTTGTTTGGTTCTCT530CAG-TAGATTCTCTTGATCTAGATCTTCCCACGAGCCATAGTTTTTTTTTCTTTTTGCTCAAAGTTTGAGATCTTAAGAATGGAAGATCTAGATCAAG531CTTGATCTAGATCTTCC532Male-sterileAAGATCTCAAACTTTGAGCAAAAAGAAAAAAAAACTATGGCTCGTG533AP3GGAAGATCCAGATCTAGAGAATAGAGAACCAAACAAACAGACAAGNicotiana tabacumTCACTTATTCTAAGAGAAGAAATGGACTTTLys9TermAAAGTCCATTTCTTCTCTTAGAATAAGTGACTTGTCTGTTTGTTTGG534AAG-TAGTTCTCTATTCTCTAGATCTGGATCTTCCCACGAGCCATAGTTTTTTTTTCTTTTTGCTCAAAGTTTGAGATCTTTCCAGATCTAGAGAATA535TATTCTCT+E,un AGATCTGGA536Male-sterileATCTCAAACTTTGAGCAAAAAGAAAAAAAAACTATGGCTCGTGGGA537AP3AGATCCAGATCAAGTGAATAGAGAACCAAACAAACAGACAAGTCANicotiana tabacumCTTATTCTAAGAGAAGAAATGGACTTTTCAArg10TermTGAAAAGTCCATTTCTTCTCTTAGAATAAGTGACTTGTCTGTTTGTT538AGA-TGATGGTTCTCTATTCACTTGATCTGGATCTTCCCACGAGCCATAGTTTTTTTTTCTTTTTGCTCAAAGTTTGAGATAGATCAAGTGAATAGAG539CTCTATTCACTTGATCT540Male-sterileGGCTCGAGGAAAGATCCAGATCAAGAGAATAGAGAACACAACGAA541AP3CAGACAAGTAACTTAGTCAAAACGAAGGGATGGTCTTTTCAAGAAGMedicago sativaGCCAATGAGCTCACTGTTCTTTGTGATGCTTyr21TermAGCATCACAAAGAACAGTGAGCTCATTGGCCTTCTTGAAAAGACCA542TAC-TAGTCCCTTCGTTTTGACTAAGTTACTTGTCTGTTCGTTGTGTTCTCTATTCTCTTGATCTGGATCTTTCCTCGAGCCGTAACTTAGTCAAAACG543CGTTTTGACTAAGTTAC544Male-sterileCTCGAGGAAAGATCCAGATCAAGAGAATAGAGAACACAACGAACA545AP3GACAAGTAACTTACTGAAAACGAAGGGATGGTCTTTTCAAGAAGGMedicago sativaCCAATGAGCTCACTGTTCTTTGTGATGCTAASer22TermTTAGCATCACAAAGAACAGTGAGCTCATTGGCCTTCTTGAAAAGAC546TCA-TGACATCCCTTCGTTTTCAGTAAGTTACTTGTCTGTTCGTTGTGTTCTCTATTCTCTTGATCTGGATCTTTCCTCGAGAACTTACTGAAAACGAA547TTCGTTTT+E,un CAGTAAGTT548Male-sterileCGAGGAAAGATCCAGATCAAGAGAATAGAGAACACAACGAACAGA549AP3CAAGTAACTTACTCATAACGAAGGGATGGTCTTTTCAAGAAGGCCAMedicago sativaATGAGCTCACTGTTCTTTGTGATGCTAAGGLys23TermCCTTAGCATCACAAAGAACAGTGAGCTCATTGGCCTTCTTGAAAAG550AAA-TAAACCATCCCTTCGTTATGAGTAAGTTACTTGTCTGTTCGTTGTGTTCTCTATTCTCTTGATCTGGATCTTTCCTCGCTTACTCATAACGAAGG551CCTTCGTTATGAGTAAG552Male-sterileGGAAAGATCCAGATCAAGAGAATAGAGAACACAACGAACAGACAA553AP3GTAACTTACTCAAAATGAAGGGATGGTCTTTTCAAGAAGGCCAATGMedicago sativaAGCTCACTGTTCTTTGTGATGCTAAGGTTTArg24TermAAACCTTAGCATCACAAAGAACAGTGAGCTCATTGGCCTTCTTGAA554CGA-TGAAAGACCATCCCTTCATTTTGAGTAAGTTACTTGTCTGTTCGTTGTGTTCTCTATTCTCTTGATCTGGATCTTTCCACTCAAAATGAAGGGAT555ATCCCTTCATTTTGAGT556Male-sterileGGCTCGTGGTAAGATCCAGATCAAGAAAATAGAAAACCAAACAAAT557DEF4AGGCAAGTGACTTAGTCAAAGAGAAGAAATGGGCTATTCAAGAAGSolanum tuberosumGCTAATGAACTTACAGTTCTTTGTGATGCTTyr21TermAGCATCACAAAGAACTGTAAGTTCATTAGCCTTCTTGAATAGCCCA558TAT-TAGTTTCTTCTCTTTGACTAAGTCACTTGCCTATTTGTTTGGTTTTCTATTTTCTTGATCTGGATCTTACCACGAGCCGTGACTTAGTCAAAGAG559CTCTTTGACTAAGTCAC560Male-sterileCTCGTGGTAAGATCCAGATCAAGAAAATAGAAAACCAAACAAATAG561DEF4GCAAGTGACTTATTGAAAGAGAAGAAATGGGCTATTCAAGAAGGCSolanum tuberosumTAATGAACTTACAGTTCTTTGTGATGCTAASer22TermTTAGCATCACAAAGAACTGTAAGTTCATTAGCCTTCTTGAATAGCC562TCA-TGACATTTCTTCTCTTTCAATAAGTCACTTGCCTATTTGTTTGGTTTTCTATTTTCTTGATCTGGATCTTACCACGAGGACTTATTGAAAGAGAA563TTCTCTTTCAATAAGTC564Male-sterileCGTGGTAAGATCCAGATCAAGAAAATAGAAAACCAAACAAATAGG565DEF4CAAGTGACTTATTCATAGAGAAGAAATGGGCTATTCAAGAAGGCTASolanum tuberosumATGAACTTACAGTTCTTTGTGATGCTAAAGLys23TermCTTTAGCATCACAAAGAACTGTAAGTTCATTAGCCTTCTTGAATAG566AAG-TAGCCCATTTCTTCTCTATGAATAAGTCACTTGCCTATTTGTTTGGTTTTCTATTTTCTTGATCTGGATCTTACCACGCTTATTCATAGAGAAGA567TCTTCTCTATGAATAAG568Male-sterileGGTAAGATCCAGATCAAGAAAATAGAAAACCAAACAAATAGGCAA569DEF4GTGACTTATTCAAAGTGAAGAAATGGGCTATTCAAGAAGGCTAATGSolanum tuberosumAACTTACAGTTCTTTGTGATGCTAAAGTTTArg24TermAAACTTTAGCATCACAAAGAACTGTAAGTTCATTAGCCTTCTTGAAT570AGA-TGAAGCCCATTTCTTCACTTTGAATAAGTCACTTGCCTATTTGTTTGGTTTTCTATTTTCTTGATCTGGATCTTACCATTCAAAGTGAAGAAAT571ATTTCTTCAGTTTGAAT572Male-sterileGCTAATGAACTTACTGTTCTTTGTGATGCTAAAGTTTCAATTGTTAT573AP3GATTTCTAGTACTTGAAAACTTCATGAGTTTATAAGTCCCTCTATCALycopersiconCGACCAAACAATTGTTCGATCTGTACCesculentumGGTACAGATCGAACAATTGTTTGGTCGTGATAGAGGGACTTATAAA574Gly27TermCTCATGAAGTTTTCAAGTACTAGAAATCATAACAATTGAAACTTTAGGGA-TGACATCACAAAGAACAGTAAGTTCATTAGCCTAGTACTTGAAAACTT575AAGTTTTCAAGTACTAG576Male-sterileAATGAACTTACTGTTCTTTGTGATGCTAAAGTTTCAATTGTTATGAT577AP3TTCTAGTACTGGATAACTTCATGAGTTTATAAGTCCCTCTATCACGALycopersiconCCAAACAATTGTTCGATCTGTACCAGAesculentumTCTGGTACAGATCGAACAATTGTTTGGTCGTGATAGAGGGACTTAT578Lys28TermAAACTCATGAAGTTATCCAGTACTAGAAATCATAACAATTGAAACTTAAA-TAATAGCATCACAAAGAACAGTAAGTTCATTGTACTGGATAACTTCAT579ATGAAGTTATCCAGTAC580Male-sterileACTGTTCTTTGTGATGCTAAAGTTTCAATTGTTATGATTTCTAGTAC581AP3TGGAAAACTTCATTAGTTTATAAGTCCCTCTATCACGACCAAACAATLycopersiconTGTTCGATCTGTACCAGAAGACTATTGesculentumCAATAGTCTTCTGGTACAGATCGAACAATTGTTTGGTCGTGATAGA582Glu31TermGGGACTTATAAACTAATGAAGTTTTCCAGTACTAGAAATCATAACAGAG-TAGATTGAAACTTTAGCATCACAAAGAACAGTAACTTCATTAGTTTATA583TATAAACTAATGAAGTT584Male-sterileATTGTTATGATTTCTAGTACTGGAAAACTTCATGAGTTTATAAGTCC585AP3CTCTATCACGACCTAACAATTGTTCGATCTGTACCAGAAGACTATTLycopersiconGGAGTTGATATTTGGACTACTCACTATGesculentumCATAGTGAGTAGTCCAAATATCAACTCCAATAGTCTTCTGGTACAG586Lys40TermATCGAACAATTGTTAGGTCGTGATAGAGGGACTTATAAACTCATGAAAA-TAAAGTTTTCCAGTACTAGAAATCATAACAATTCACGACCTAACAATTG587CAATTGTTAGGTCGTGA588Male-sterileGGGGCGGGGGAAGATTGAGATAAAGCGGATCGAGAACGCCACCA589AP3ACAGGCAGGTGACCTAGTCCAAGCGCCGGTCGGGGATCATGAAGTriticum aestivumAAGGCGCGGGAGCTCACCGTGCTCTGCGACGCCTyr21TermGGCGTCGCAGAGCACGGTGAGCTCCCGCGCCTTCTTCATGATCC590TAC-TAGCCGACCGGCGCTTGGACTAGGTCACCTGCCTGTTGGTGGCGTTCTCGATCCGCTTTATCTCAATCTTCCCCCGCCCCGTGACCTAGTCCAAGCG591CGCTTGGACTAGGTCAC592Male-sterileCGGGGGAAGATTGAGATAAAGCGGATCGAGAACGCCACCAACAG593AP3GCAGGTGACCTACTCCTAGCGCCGGTCGGGGATCATGAAGAAGGTriticum aestivumCGCGGGAGCTCACCGTGCTCTGCGACGCCCAGGLys23TermCCTGGGCGTCGCAGAGCACGGTGAGCTCCCGCGCCTTCTTCATG594AAG-TAGATCCCCGACCGGCGCTAGGAGTAGGTCACCTGCCTGTTGGTGGCGTTCTCGATCCGCTTTATCTCAATCTTCCCCCGCCTACTCCTAGCGCCGG595CCGGCGCTAGGAGTAGG596Male-sterileTTGAGATAAAGCGGATCGAGAACGCCACCAACAGGCAGGTGACCT597AP3ACTCGAAGCGCCGGTAGGGGATCATGAAGAAGGCGCGGGAGCTCTriticum aestivumACCGTGCTCTGCGACGCCCAGGTCGCCATCATSer26TermATGATGGCGACCTGGGCGTCGCAGAGCACGGTGAGCTCCCGCGC598TCG-TAGCTTCTTCATGATCCCCTACCGGCGCTTGGAGTAGGTCACCTGCCTGTTGGTGGCGTTGTCGATCCGCTTTATCTCAAGCGCCGGTAGGGGATCA599TGATCCCCTACCGGCGC600Male-sterileCGGATCGAGAACGCCACCAACAGGCAGGTGACCTACTCCAAGCG601AP3CCGGTCGGGGATCATGTAGAAGGCGCGGGAGCTCACCGTGCTCTTriticum aestivumGCGACGCCCAGGTCGCCATCATCATGTTCTCCTLys30TermAGGAGAACATGATGATGGCGACCTGGGCGTCGCAGAGCACGGTG602AAG-TAGAGCTCCCGCGCCTTCTACATGATCCCCGACCGGCGCTTGGAGTAGGTCACCTGCCTGTTGGTGGCGTTGTCGATCCGGGATCATGTAGAAGGCG603CGCCTTCTACATGATCC604Male-sterileGGGGCGCGGCAAGATCGAGATCAAGCGGATCGAGAACGCCACCA605Silky1ACCGCCAGGTGACCTAGTCCAAGCGCCGGACGGGGATCATGAAGZea maysAAGGCACGCGAGCTCACCGTGCTCTGCGACGCCTyr21TermGGCGTCGCAGAGCACGGTGAGCTCGCGTGCCTTCTTCATGATCCC606TAG-TAGCGTCCGGCGCTTGGACTAGGTCACCTGGCGGTTGGTGGCGTTCTCGATCGGCTTGATCTCGATCTTGCCGCGCCCCGTGACCTAGTCCAAGCG607CGCTTGGACTAGGTCAC608Male-sterileCGCGGCAAGATCGAGATCAAGCGGATCGAGAACGCCACCAACCG609Silky1CCAGGTGACCTACTCCTAGCGCCGGACGGGGATCATGAAGAAGGZea maysCACGCGAGCTCACCGTGCTCTGCGACGCCCAGGLys23TermCCTGGGCGTCGCAGAGCACGGTGAGCTCGCGTGCCTTCTTCATG610AAG-TAGATCCCCGTCCGGCGCTAGGAGTAGGTCACCTGGCGGTTGGTGGCGTTCTCGATCCGCTTGATCTCGATCTTGCCGCGCCTACTCCTAGCGCCGG611CCGGCGCTAGGAGTAGG612Male-sterileCGGATCGAGAACGCCACCAACCGCCAGGTGACCTACTCCAAGCG613Silky1CCGGACGGGGATCATGTAGAAGGCACGCGAGCTCACCGTGCTCTZea maysGCGACGCCCAGGTCGCCATCATCATGTTCTCCTLys30TermAGGAGAACATGATGATGGCGACCTGGGCGTCGCAGAGCACGGTG614AAG-TAGAGCTCGCGTGCCTTCTACATGATCCCGGTCCGGCGCTTGGAGTAGGTCACCTGGCGGTTGGTGGCGTTCTCGATCCGGGATCATGTAGAAGGCA615TGCCTTCTACATGATCC616Male-sterileATCGAGAACGCCACCAACCGCCAGGTGACGTACTCCAAGCGCCG617Silky1GACGGGGATCATGAAGTAGGCACGCGAGCTCACCGTGCTCTGCGZea maysACGCCCAGGTCGCCATCATCATGTTCTCCTCCALys31TermTGGAGGAGAACATGATGATGGCGACCTGGGCGTCGCAGAGCACG618AAG-TAGGTGAGCTCGCGTGCCTACTTCATGATCCCCGTCCGGCGCTTGGAGTAGGTCACCTGGCGGTTGGTGGCGTTCTCGATTCATGAAGTAGGCACGC619GCGTGCCTACTTCATGA620Male-sterileGCTAGCTGCATTGTCCGGCGAGAGAGATAGCTGCTGCAGGGGGC621AP3GGCCATGGGGAGGGGCTAGATCGAGATCAAGCGGATCGAGAACGOryza sativaCGACCAACAGGCAGGTGACCTACTCGAAGCGCCLys5TermGGCGCTTGGAGTAGGTCACCTGCCTGTTGGTCGCGTTCTCGATCC622AAG-TAGGCTTGATCTCGATCTAGCCCGTCCCCATGGCGGCCCCCTGCAGCAGCTATCTCTCTCGCCGGACAATGCAGCTAGCGGAGGGGCTAGATCGAG623CTCGATCTAGCCCCTCC624Male-sterileTGCATTGTCCGGCGAGAGAGATAGCTGCTGCAGGGGGCGGCCAT625AP3GGGGAGGGGCAAGATCTAGATCAAGCGGATCGAGAACGCGACCAOryza sativaACAGGCAGGTGACCTACTCGAAGCGCCGCACGGGlu7TermCCGTGCGGCGCTTCGAGTAGGTCACCTGCCTGTTGGTCGCGTTCT626GAG-TAGCGATCCGCTTGATCTAGATCTTGCCCCTCCCCATGGCCGCCCCCTGCAGCAGCTATCTCTCTCGCCGGACAATGCAGCAAGATCTAGATCAAG627CTTGATCTAGATCTTGC628Male-sterileGTCCGGCGAGAGAGATAGCTGCTGCAGGGGGCGGCCATGGGGA629AP3GGGGCAAGATCGAGATCTAGCGGATCGAGAACGCGACCAACAGGOryza sativaCAGGTGACCTACTCGAAGCGCCGCACGGGGATCALys9TermTGATCCCCGTGCGGCGCTTCGAGTAGGTCACCTGCCTGTTGGTCG630AAG-TAGCGTTCTCGATCCGCTAGATCTCGATCTTGCCCCTCCCCATGGCCGCCCCCTGCAGCAGCTATCTCTCTCGCCGGACTCGAGATCTAGCGGATC631GATCCGCTAGATCTCGA632Male-sterileGAGAGATAGCTGCTGCAGGGGGCGGCCATGGGGAGGGGCAAGA633AP3TCGAGATCAAGCGGATCTAGAACGCGACCAACAGGCAGGTGACCTOryza sativaACTCGAAGCGCCGCACGGGGATCATGAAGAAGGGlu12TermCCTTCTTCATGATCCCCGTGCGGCGCTTCGAGTAGGTCACCTGCC634GAG-TAGTGTTGGTCGCGTTCTAGATCCGCTTGATCTCGATCTTGCCCCTCCCCATGGCGGCCCCCTGCAGCAGCTATCTCTCAGCGGATCTAGAACGCG635CGCGTTCTAGATCCGCT636

[0126]

17

TABLE 15

Oligonucleotides to produce male-sterile plants

Phenotype, Gene,

Plant & Targeted

SEQ ID

Alteration
Altering Oligos
NO:

Male-sterile
TCTGTACTAATCAAATTTTGCCCTAAACGTTTTTGGCTTTGGAGCA
637

AG
GCAATCACGGCGTAGCAATCGGAGCTAGGAGGAGATTCCTCTCC

Arabidopsis thaliana

CTTGAGGAAATCTGGGAGAGGAAAGATCGAA

Tyr35Term
TTCGATCTTTCCTCTCCCAGATTTCCTCAAGGGAGAGGAATCTCCT
638

TAG-TAG
CCTAGGTCCGATTGCTACGCCGTGATTGCTGCTCCAAAGCCAAAA

ACGTTTAGGGCAAAATTTGATTAGTACAGA

ACGGCGTAGCAATCGGA
639

TCCGATTGCTACGCCGT
640

Male-sterile
CTGTACTAATCAAATTTTGCCCTAAACGTTTTTGGCTTTGGAGCAG
641

AG
CAATCACGGCGTACTAATCGGAGCTAGGAGGAGATTCCTCTCCCT

Arabidopsis thaliana

TGAGGAAATCTGGGAGAGGAAAGATCGAAA

Gln36Term
TTTCGATCTTTCCTCTCCCAGATTTCCTCAAGGGAGAGGAATCTCC
642

CAA-TAA
TCCTAGCTCCGATTAGTACGCCGTGATTGCTGCTCCAAAGCCAAA

AACGTTTAGGGCAAAATTTGATTAGTACAG

CGGCGTACTAATCGGAG
643

CTCCGATTAGTACGCCG
644

Male-sterile
ACTAATCAAATTTTGCCCTAAACGTTTTTGGCTTTGGAGCAGCAAT
645

AG
CACGGCGTACCAATAGGAGCTAGGAGGAGATTCCTCTCCCTTGA

Arabidopsis thaliana

GGAAATCTGGGAGAGGAAAGATCGAAATCAA

Ser37Term
TTGATTTCGATCTTTCCTCTCCCAGATTTCCTCAAGGGAGAGGAAT
646

TCG-TAG
CTCCTCCTAGCTCCTATTGGTACGCCGTGATTGCTGCTCCAAAGC

CAAAAACGTTTAGGGCAAAATTTGATTAGT

GTACCAATAGGAGCTAG
647

CTAGCTCCTATTGGTAC
648

Male-sterile
TAATCAAATTTTGCCCTAAACGTTTTTGGCTTTGGAGCAGCAATCA
649

AG
CGGCGTACCAATCGTAGCTAGGAGGAGATTCCTCTCCCTTGAGGA

Arabidopsis thalana

AATCTGGGAGAGGAAAGATCGAAATCAAAC

Glu38Term
GTTTGATTTCGATCTTTCCTCTCCCAGATTTCCTCAAGGGAGAGGA
650

GAG-TAG
ATCTCCTCCTAGCTACGATTGGTACGCCGTGATTGCTGCTCCAAA

GCCAAAAACGTTTAGGGCAAAATTTGATTA

ACCAATCGTAGCTAGGA
651

TCCTAGCTACGATTGGT
652

Male-sterile
CTCTCCCACTTCTTTTCGGTGGTTTATTCATTTGGTGACGATATCA
653

AG
CAGAAGCAATGGATTAAGGTGGGAGTAGTCACGATGCAGAGAGT

Brassica napus

AGCAAGAAGATAGGTAGAGGGAAGATAGAGA

Glu3Term
TCTCTATCTTCCCTCTACCTATCTTCTTGCTACTCTCTGCATCGTGA
654

GAA-TAA
CTACTCCCACCTTAATCCATTGCTTCTGTGATATCGTCACCAAATG

AATAAACCACCGAAAAGAAGTGGGAGAG

CAATGGATTAAGGTGGG
655

CCCACCTTAATCCATTG
656

Male-sterile
TATTCATTTGGTGACGATATCACAGAAGCAATGGATGAAGGTGGG
657

AG
AGTAGTCACGATGCATAGAGTAGCAAGAAGATAGGTAGAGGGAA

Brassica napus

GATAGAGATAAAGAGGATAGAGAACACAACAA

Glu11Term
TTGTTGTGTTCTCTATCCTCTTTATCTCTATCTTCCCTCTACCTATC
658

GAG-TAG
TTCTTGCTACTCTATGCATCGTGACTACTCCCACCTTCATCCATTG

CTTCTGTGATATCGTCACCAAATGAATA

ACGATGCATAGAGTAGC
659

GCTACTCTATGCATCGT
660

Male-sterile
GGTGACGATATCACAGAAGCAATGGATGAAGGTGGGAGTAGTCA
661

AG
CGATGCAGAGAGTAGCTAGAAGATAGGTAGAGGGAAGATAGAGA

Brassica napus

TAAAGAGGATAGAGAACACAACAAATCGTCAAG

Lys14Term
CTTGACGATTTGTTGTGTTCTCTATCCTCTTTATCTCTATCTTCCCT
662

AAG-TAG
GTACCTATCTTCTAGCTACTCTCTGCATCGTGACTACTCCCACCTT

CATCCATTGCTTCTGTGATATCGTCACC

AGAGTAGCTAGAAGATA
663

TATCTTCTAGCTAGTCT
664

Male-sterile
GACGATATCACAGAAGCAATGGATGAAGGTGGGAGTAGTCACGA
665

AG
TGCAGAGAGTAGCAAGTAGATAGGTAGAGGGAAGATAGAGATAAA

Brassica napus

GAGGATAGAGAACACAACAAATCGTCAAGTAA

Lys15Term
TTACTTGACGATTTGTTGTGTTCTCTATCCTCTTTATCTCTATCTTC
666

AAG-TAG
CCTCTACCTATCTACTTGCTACTCTCTGCATCGTGACTACTCCCAC

CTTCATCCATTGCTTCTGTGATATCGTC

GTAGCAAGTAGATAGGT
667

ACCTATCTACTTGCTAC
668

Male-sterile
CAACCAAAAAACTTAAAAATCTTCTCTTTCCTTTCCTTACAAGGTGA
669

AG
AGTAATGGACTTCTAAAGTGATCTAACCAGAGAGATCTCACCACAA

Lycopersicon

AGGAAACTAGGAAGGGGGAAAATTGAGA

esculentum

TCTCAATTTTCCCCCTTCCTAGTTTCCTTTGTGGTGAGATCTCTCT
670

Glu4Term
GGTTAGATCACTTTAGAAGTCCATTACTTCACCTTGTAAGGAAAGG

CAA-TAA
AAAGAGAAGATTTTTAAGTTTTTTGGTTG

TGGACTTC+E,unc TAAAGTGAT
671

ATCACTTTAGAAGTCCA
672

Male-sterile
AAAATCTTCTCTTTCCTTTCCTTACAAGGTGAAGTAATGGACTTCC
673

AG
AAAGTGATCTAACCTGAGAGATCTCACCACAAAGGAAACTAGGAA

Lycopersicon

GGGGGAAAATTGAGATCAAAAGGATCGAAA

esculentum

TTTCGATCCTTTTGATCTCAATTTTCCCCCTTCCTAGTTTCCTTTGT
674

Arg9Term
GGTGAGATCTCTCAGGTTAGATCACTTTGGAAGTCCATTACTTCAC

AGA-TGA
CTTGTAAGGAAAGGAAAGAGAAGATTTT

ATCTAACCTGAGAGATC
675

GATCTCTCAGGTTAGAT
676

Male-sterile
ATCTTCTCTTTCCTTTCCTTACAAGGTGAAGTAATGGACTTCCAAA
677

AG
GTGATCTAACCAGATAGATCTCACCACAAAGGAAACTAGGAAGGG

Lycopersicon

GGAAAATTGAGATCAAAAGGATCGAAAACA

esculentum

TGTTTTCGATCCTTTTGATCTCAATTTTCCCCCTTCCTAGTTTCCTT
678

Glu10Term
TGTGGTGAGATCTATCTGGTTAGATCACTTTGGAAGTCCATTACTT

GAG-TAG
CACCTTGTAAGGAAAGGAAAGAGAAGAT

TAACCAGATAGATCTCA
679

TGAGATCTATCTGGTTA
680

Male-sterile
CTTTCCTTTCCTTACAAGGTGAAGTAATGGACTTCCAAAGTGATCT
681

AG
AACCAGAGAGATCTGACCACAAAGGAAACTAGGAAGGGGGAAAA

Lycopersicon

TTGAGATCAAAAGGATCGAAAACACGACGAA

esculentum

TTCGTCGTGTTTTCGATCCTTTTGATCTCAATTTTCCCCCTTCCTAG
682

Ser12Term
TTTCCTTTGTGGTCAGATCTCTGTGGTTAGATCACTTTGGAAGTCC

TCA-TGA
ATTACTTCACCTTGTAAGGAAAGGAAAG

AGAGATCTGACCACAAA
683

TTTGTGGTCAGATCTCT
684

Male-sterile
GTACTCTCTATTTTCATCTTCCAACCCTTTCTTTCCTTACCAGGTGA
685

NAG1
AAGTATGGACTTCTAAAGTGATCTAACAAGAGAGATCTCTCCACAA

Nicotiana tabacum

AGGAAACTGGGAAGAGGAAAGATTGAGA

Gln4Term
TCTCAATCTTTCCTCTTCCCAGTTTCCTTTGTGGAGAGATCTCTCTT
686

CAA-TAA
GTTAGATCACTTTAGAAGTCCATACTTTCACCTGGTAAGGAAAGAA

AGGGTTGGAAGATGAAAATAGAGAGTAC

TGGACTTCTAAAGTGAT
687

ATCACTTTAGAAGTCCA
688

Male-sterile
ATCTTCCAACCCTTTCTTTCCTTACCAGGTGAAAGTATGGACTTCC
689

NAG1
AAAGTGATCTAACATGAGAGATCTCTCCACAAAGGAAACTGGGAA

Nicotiana tabacum

GAGGAAAGATTGAGATCAAACGGATCGAAA

Arg9Term
TTTCGATCCGTTTGATCTCAATCTTTCCTCTTCCCAGTTTCCTTTGT
690

AGA-TGA
GGAGAGATCTCTCATGTTAGATCACTTTGGAAGTCCATACTTTCAC

CTGGTAAGGAAAGAAAGGGTTGGAAGAT

ATCTAACATGAGAGATC
691

GATCTCTCATGTTAGAT
692

Male-sterile
TTCCAACCCTTTCTTTCCTTAGCAGGTGAAAGTATGGACTTCCAAA
693

NAG1
GTGATCTAACAAGATAGATCTCTCCACAAAGGAAACTGGGAAGAG

Nicotiana tabacum

GAAAGATTGAGATCAAACGGATCGAAAACA

Glu10Term
TGTTTTCGATCCGTTTGATCTCAATCTTTCCTCTTCCCAGTTTCCTT
694

GAG-TAG
TGTGGAGAGATCTATCTTGTTAGATGACTTTGGAAGTCCATACTTT

CACCTGGTAAGGAAAGAAAGGGTTGGAA

TAACAAGATAGATCTCT
695

AGAGATCTATCTTGTTA
696

Male-sterile
CTTTCCTTACCAGGTGAAAGTATGGACTTCCAAAGTGATCTAACAA
697

NAG1
GAGAGATCTCTCCATAAAGGAAACTGGGAAGAGGAAAGATTGAGA

Nicotiana tabacum

TCAAACGGATCGAAAACACAACGAATCGTC

Gln14Term
GACGATTCGTTGTGTTTTCGATCCGTTTGATCTCAATCTTTCCTCTT
698

CAA-TAA
CCCAGTTTCCTTTATGGAGAGATCTCTCTTGTTAGATCACTTTGGA

AGTCCATACTTTCACCTGGTAAGGAAAG

TCTCTCCATAAAGGAAA
699

TTTCCTTTATGGAGAGA
700

Male-sterile
GCCTATGAAAACAAACCCAACACGGTCCTGGACGCTGATGCCCAA
701

AG
AGAAGATTGGGAAGGTGAAAGATCGAGATCAAGCGGATCGAAAA

Rosa hybrida

CACCACCAATCGTCAAGTCACCTTCTGCAAAA

Gly22Term
TTTTGCAGAAGGTGACTTGACGATTGGTGGTGTTTTCGATCCGCT
702

GGA-TGA
TGATCTGGATCTTTCACCTTCCCAATCTTCTTTGGGCATCAGCGTC

CAGGACCGTGTTGGGTTTGTTTTCATAGGC

TGGGAAGGTGAAAGATC
703

GATCTTTCACCTTCCCA
704

Male-sterile
TATGAAAACAAACCCAACACGGTCCTGGACGCTGATGCCCAAAGA
705

AG
AGATTGGGAAGGGGATAGATCGAGATCAAGCGGATCGAAAACAC

Rosa hybrida

CACCAATCGTCAAGTCACCTTCTGCAAAAGGC

Lys23Term
GCCTTTTGCAGAAGGTGACTTGACGATTGGTGGTGTTTTCGATCC
706

AAG-TAG
GCTTGATCTCGATCTATCCCCTTCCCAATCTTCTTTGGGCATCAGC

GTCCAGGACCGTGTTGGGTTTGTTTTCATA

GAAGGGGATAGATCGAG
707

CTCGATCTATCCCCTTC
708

Male-sterile
AACAAACCCAACACGGTCCTGGACGCTGATGCCCAAAGAAGATTG
709

AG
GGAAGGGGAAAGATCTAGATCAAGCGGATCGAAAACACCACCAA

Rosa hybrida

TCGTCAAGTCACCTTCTGCAAAAGGCGCAATG

Glu25Term
CATTGCGCCTTTTGCAGAAGGTGACTTGACGATTGGTGGTGTTTT
710

GAG-TAG
CGATCCGCTTGATCTAGATCTTTCCCCTTCCCAATCTTCTTTGGGC

ATCAGCGTCCAGGACCGTGTTGGGTTTGTT

GAAAGATCTAGATCAAG
711

CTTGATCTAGATCTTTC
712

Male-sterile
CCCAACACGGTCCTGGACGCTGATGCCCAAAGAAGATTGGGAAG
713

AG
GGGAAAGATCGAGATCTAGCGGATCGAAAACACCACCAATCGTCA

Rosa hybrida

AGTCACCTTCTGCAAAAGGCGCAATGGTTTGC

Lys27
GCAAACCATTGCGCCTTTTGCAGAAGGTGACTTGACGATTGGTGG
714

AAG-TAG
TGTTTTCGATCCGCTAGATCTCGATCTTTCCCCTTCCCAATCTTCT

TTGGGCATCAGCGTCCAGGACCGTGTTGGG

TCGAGATCTAGCGGATC
715

GATCCGCTAGATCTCGA
716

Male-sterile
CAATTGCGTGTTTTTATTTTTTTTGTTTTTGACTAAGTAGAAATGGC
717

far
GTCTCTAAGCGATTAATCGACCGAGGTATCGCGCGAGAGGAAAAT

Antirrhinum majus

CGGGAGAGGAAAGATCGAGATCAAACGGA

Gln7Term
TCCGTTTGATCTCGATCTTTCCTCTCCCGATTTTCCTCTCGGGCGA
718

CAA-TAA
TACCTCGGTCGATTAATCGCTTAGAGACGCCATTTCTACTTAGTCA

AAAAGAAAAAAAATAAAAACAGGCAATTG

TAAGCGATTAATCGACC
719

GGTCGATTAATCGCTTA
720

Male-sterile
GTTTTTATTTTTTTTCTTTTTGACTAAGTAGAAATGGCGTCTCTAAG
721

far
CGATCAATCGACCTAGGTATCGCCCGAGAGGAAAATCGGGAGAG

Antirrhinum majus

GAAAGATCGAGATCAAACGGATCGAAAACA

Glu10Term
TGTTTTCGATCCGTTTGATCTCGATCTTTCCTCTCCCGATTTTCCTC
722

GAG-TAG
TCGGGCGATACCTAGGTCGATTGATCGCTTAGAGACGCCATTTCT

ACTTAGTCAAAAAGAAAAAAAATAAAAAC

AATCGACCTAGGTATCG
723

CGATACCTAGGTCGATT
724

Male-sterile
TTTCTTTTTGACTAAGTAGAAATGGCGTCTCTAAGCGATCAATCGA
725

far
CCGAGGTATCGCCCTAGAGGAAAATCGGGAGAGGAAAGATCGAG

Antirrhinum majus

ATCAAACGGATCGAAAACAAAACAAATCAAC

Glu14Term
GTTGATTTGTTTTGTTTTCGATCCGTTTGATCTCGATCTTTCCTCTC
726

GAG-TAG
CCGATTTTCCTCTAGGGCGATACCTCGGTCGATTGATCGCTTAGA

GACGCCATTTCTACTTAGTCAAAAAGAAA

TATCGCCCTAGAGGAAA
727

TTTCCTCTAGGGCGATA
728

Male-sterile
TTTGACTAAGTAGAAATGGCGTCTCTAAGCGATCAATCGACCGAG
729

far
GTATCGCCCGAGAGGTAAATCGGGAGAGGAAAGATCGAGATCAA

Antirrhinum majus

ACGGATCGAAAACAAAACAAATCAACAGGTTA

Lys16Term
TAACCTGTTGATTTGTTTTGTTTTCGATCCGTTTGATCTCGATCTTT
730

AAA-TAA
CCTCTCCCGATTTACCTCTCGGGCGATACCTCGGTCGATTGATCG

CTTAGAGACGCCATTTCTACTTAGTCAAA

CCGAGAGGTAAATCGGG
731

CCCGATTTACCTCTCGG
732

Male-sterile
TGTCCAAGCATTATCAGTCACCACTCACAAGAATGATTAAGGAAGA
733

AG
AGGAAAGGGTAAGTAGCAAATAAAGGGGATGTTCCAGAATCAAGA

Cucumis sativus

AGAGAAGATGTCAGACTCGCCTCAGAGGAA

Leu21Term
TTCCTCTGAGGCGAGTCTGACATCTTCTCTTCTTGATTCTGGAACA
734

TTG-TAG
TCCCCTTTATTTGCTACTTACCCTTTCCTTCTTCCTTAATCATTCTT

GTGAGTGGTGACTGATAATGCTTGGACA

GGGTAAGTAGCAAATAA
735

TTATTTGCTACTTACCC
736

Male-sterile
TCCAAGCATTATCAGTCACCACTCACAAGAATGATTAAGGAAGAA
737

AG
GGAAAGGGTAAGTTGTAAATAAAGGGGATGTTCCAGAATCAAGAA

Cucumis sativus

GAGAAGATGTCAGACTCGCCTCAGAGGAAGA

Gln22Term
TCTTCCTCTGAGGCGAGTCTGACATCTTCTCTTCTTGATTCTGGAA
738

CAA-TAA
CATCCCCTTTATTTACAACTTACCCTTTCCTTCTTCCTTAATCATTC

TTGTGAGTGGTGACTGATAATGCTTGGA

GTAAGTTGTAAATAAAG
739

CTTTATTTACAACTTAC
740

Male-sterile
CATTATCAGTCACCACTCACAAGAATGATTAAGGAAGAAGGAAAG
741

AG
GGTAAGTTGCAAATATAGGGGATGTTCCAGAATCAAGAAGAGAAG

Cucumis sativus

ATGTCAGACTCGCCTCAGAGGAAGATGGGAA

Lys24Term
TTCCCATCTTCCTCTGAGGCGAGTCTGACATCTTCTCTTCTTGATT
742

AAG-TAG
CTGGAACATCCCCTATATTTGCAACTTACCCTTTCCTTCTTCCTTAA

TCATTCTTGTGAGTGGTGACTGATAATG

TGCAAATATAGGGGATG
743

CATCCCCTATATTTGCA
744

Male-sterile
CCACTCACAAGAATGATTAAGGAAGAAGGAAAGGGTAAGTTGCAA
745

AG
ATAAAGGGGATGTTCTAGAATCAAGAAGAGAAGATGTCAGACTCG

Cucumis sativus

CCTCAGAGGAAGATGGGAAGAGGAAAGATTG

Gln28Term
CAATCTTTCCTCTTCCCATCTTCCTCTGAGGCGAGTCTGACATCTT
746

CAG-TAG
CTCTTCTTGATTCTAGAACATCCCCTTTATTTGCAACTTACCCTTTC

CTTCTTCCTTAATCATTCTTGTGAGTGG

GGATGTTCTAGAATCAA
747

TTGATTCTAGAACATCC
748

Male-sterile
CCACCACCACCACCACCACCACCACCACACCATGCTCAACATGAT
749

AG
GACTGATCTGAGCTGAGGGCCGTCGTCCAAGGTCAAGGAGCAGG

Zea mays

TGGCGGCGGCGCCGACGGGCTCCGGCGACAGG

Cys10Term
CCTGTCGCCGGAGCCCGTCGGCGCCGCCGCCACCTGCTCCTTGA
750

TGC-TGA
CCTTGGACGACGGCCCTCAGCTCAGATCAGTCATCATGTTGAGCA

TGGTGTGGTGGTGGTGGTGGTGGTGGTGGTGG

CTGAGCTGAGGGCCGTC
751

GACGGCCCTCAGCTCAG
752

Male-sterile
ACCACCACCACCACCACCACACCATGCTCAACATGATGACTGATC
753

AG
TGAGCTGCGGGCCGTAGTCCAAGGTCAAGGAGCAGGTGGCGGC

Zea mays

GGCGCCGACGGGCTCCGGCGACAGGCAGGGGCA

Ser13Term
TGCCCCTGCCTGTCGCCGGAGCCCGTCGGCGCCGCCGCCACCT
754

TCG-TAG
GCTCCTTGACCTTGGACTACGGCCCGCAGCTCAGATCAGTCATCA

TGTTGAGCATGGTGTGGTGGTGGTGGTGGTGGT

CGGGCCGTAGTCCAAGG
755

CCTTGGACTACGGCCCG
756

Male-sterile
CACCACCACCACCACACCATGCTCAACATGATGACTGATCTGAGC
757

AG
TGCGGGCCGTCGTCCTAGGTCAAGGAGCAGGTGGCGGCGGCGC

Zea mays

CGACGGGCTCCGGCGACAGGCAGGGGCAGGGGA

Lys15Term
TCCCCTGCCCCTGCCTGTCGCCGGAGCCCGTCGGCGCCGCCGC
758

AAG-TAG
CACCTGCTCCTTGACCTAGGACGACGGCCCGCAGCTCAGATCAG

TCATCATGTTGAGCATGGTGTGGTGGTGGTGGTG

CGTCGTCCTAGGTCAAG
759

CTTGACCTAGGACGACG
760

Male-sterile
CACCACCACACCATGCTCAACATGATGACTGATCTGAGCTGCGGG
761

AG
CCGTCGTCCAAGGTCTAGGAGCAGGTGGCGGCGGCGCCGACGG

Zea mays

GCTCCGGCGACAGGCAGGGGCAGGGGAGAGGCA

Lys17Term
TGCCTCTCCCCTGCCCCTGCCTGTCGCCGGAGCCCGTCGGCGCC
762

AAG-TAG
GCCGCCACCTGCTCCTAGACCTTGGACGACGGCCCGCAGCTCAG

ATCAGTCATCATGTTGAGCATGGTGTGGTGGTG

CCAAGGTCTAGGAGCAG
763

CTGCTCCTAGACCTTGG
764

Male-sterile
TCCTACCTTTTCTCCTTCAGACCTCAAAATCTGTGTGATAGGAACA
765

AG
AGAGCATGCACATCTGAGAAGAGGAGGCTACACCATCCACAGTAA

Zea mays

CAGGCATCATGTCGACCCTGACTTCGGCGG

Arg4Term
CCGCCGAAGTCAGGGTCGACATGATGCCTGTTACTGTGGATGGT
766

CGA-TGA
GTAGCCTCCTCTTCTCAGATGTGCATGCTCTTGTTCCTATCACACA

GATTTTGAGGTCTGAAGGAGAAAAGGTAGGA

TGCACATCTGAGAAGAG
767

CTCTTCTCAGATGTGCA
768

Male-sterile
TACCTTTTCTCCTTCAGACCTCAAAATCTGTGTGATAGGAACAAGA
769

AG
GCATGCACATCCGATAAGAGGAGGCTACACCATCCACAGTAACAG

Zea mays

GCATCATGTCGACCCTGACTTCGGCGGGGC

Glu5Term
GCCCCGCCGAAGTCAGGGTCGACATGATGCCTGTTACTGTGGAT
770

GAA-TAA
GGTGTAGCCTCCTCTTATCGGATGTGCATGCTCTTGTTCCTATCAC

ACAGATTTTGAGGTCTGAAGGAGAAAAGGTA

ACATCCGATAAGAGGAG
771

CTCCTCTTATCGGATGT
772

Male-sterile
CTTTTCTCCTTCAGACCTCAAAATCTGTGTGATAGGAACAAGAGCA
773

AG
TGCACATCCGAGAATAGGAGGCTACACCATCCACAGTAACAGGCA

Zea mays

TCATGTCGACCCTGACTTCGGCGGGGCAGC

Glu6Term
GCTGCCCCGCCGAAGTGAGGGTCGACATGATGCCTGTTACTGTG
774

GAG-TAG
GATGGTGTAGCCTCCTATTCTCGGATGTGCATGCTCTTGTTCCTAT

CACACAGATTTTGAGGTCTGAAGGAGAAAAG

TCCGAGAATAGGAGGCT
775

AGCCTCCTATTCTCGGA
776

Male-sterile
TTCTCCTTCAGACCTCAAAATCTGTGTGATAGGAACAAGAGCATG
777

AG
CACATCCGAGAAGAGTAGGCTACACCATCCACAGTAACAGGCATC

Zea mays

ATGTCGACCCTGACTTCGGCGGGGCAGCAGA

Glu7Term
TCTGCTGCCCCGCCGAAGTCAGGGTCGACATGATGCCTGTTACT
778

GAG-TAG
GTGGATGGTGTAGCCTACTCTTCTCGGATGTGCATGCTCTTGTTC

CTATCACACAGATTTTGAGGTCTGAAGGAGAA

GAGAAGAGTAGGCTACA
779

TGTAGCCTACTCTTCTC
780

Male-sterile
GCTGGGTCAGGATCGTCGGCGGCGGTGGCGGCGGGGAGCAGC
781

AG
GAGAAGATGGGGAGGGGGTAGATCGAGATAAAGCGGATCGAGAA

Oryza sativa

CACGACGAACCGGCAGGTGACCTTCTGCAAGCGCC

Lys5Term
GGCGCTTGCAGAAGGTCACCTGCCGGTTCGTCGTGTTCTCGATC
782

AAG-TAG
CGCTTTATCTCGATCTACCCCCTCCCCATCTTCTCGCTGCTCCCC

GCCGCCACCGCCGCCGACGATCCTGACCCAGC

GGAGGGGGTAGATCGAG
783

CTCGATCTACCCCCTCC
784

Male-sterile
TCAGGATCGTCGGCGGGGGTGGCGGCGGGGAGCAGCGAGAAGA
785

AG
TGGGGAGGGGGAAGATCTAGATAAAGCGGATCGAGAACACGACG

Oryza sativa

AACCGGCAGGTGACCTTCTGCAAGCGCCGCAATG

GTu7Term
CATTGCGGCGCTTGCAGAAGGTCACCTGCCGGTTCGTCGTGTTCT
786

GAG-TAG
CGATCCGCTTTATCTAGATCTTCCCCCTCCCCATCTTCTCGCTGCT

CCCCGCCGCCACCGCCGCCGACGATCCTGA

GGAAGATCTAGATAAAG
787

CTTTATCTAGATCTTCC
788

Male-sterile
TCGTCGGCGGCGGTGGCGGCGGGGAGCAGCGAGAAGATGGGG
789

AG
AGGGGGAAGATCGAGATATAGCGGATCGAGAACACGACGAACCG

Oryza sativa

GCAGGTGACCTTCTGCAAGCGCCGCAATGGCCTCC

Lys9Term
GGAGGCCATTGCGGCGCTTGCAGAAGGTCACCTGCCGGTTCGTC
790

AAG-TAG
GTGTTCTCGATCCGCTATATCTCGATCTTCCCCCTCCCCATCTTCT

CGCTGCTCCCCGCCGCCACCGCCGCCGACGA

TCGAGATATAGCGGATC
791

GATCCGCTATATCTCGA
792

Male-sterile
GCGGTGGCGGCGGGGAGCAGCGAGAAGATGGGGAGGGGGAAG
793

AG
ATCGAGATAAAGCGGATCTAGAACACGACGAACCGGCAGGTGAC

Oryza sativa

CTTCTGCAAGCGCCGCAATGGCCTCCTGAAGAAGG

Glu12Term
CCTTCTTCAGGAGGCCATTGCGGCGCTTGCAGAAGGTCACCTGC
794

GAG-TAG
CGGTTCGTCGTGTTCTAGATCCGCTTTATCTCGATCTTCCCCCTCC

CCATCTTCTCGCTGCTCCCCGCCGCCACCGC

AGCGGATCTAGAACACG
795

CGTGTTCTAGATCCGCT
796

[0127]

18

TABLE 16

Oligonucleotides to produce male-sterile plants

Phenotype, Gene,

Plant & Targeted

SEQ ID

Alteration
Altering Oligos
NO:

Male-sterile
GGGAAGAGGGAAAATAGAAATAAAAAGAATAGAGAACTCAAGCAA
797

P1
TAGACAAGTTACATAGTCAAAGAGAAGAAATGGTATCATCAAAAAA

Cucumis sativus

GCCAAAGAAATTACTGTTCTTTGCGATGCT

Tyr21Term
AGCATCGCAAAGAACAGTAATTTCTTTGGCTTTTTTGATGATACCAT
798

TAT-TAG
TTCTTCTCTTTGACTATGTAACTTGTCTATTGCTTGAGTTCTCTATTC

TTTTTATTTCTATTTTCCCTCTTCCC

GTTACATAGTCAAAGAG
799

CTCTTTGACTATGTAAC
800

Male-sterile
GAAGAGGGAAAATAGAAATAAAAAGAATAGAGAACTCAAGCAATA
801

P1
GACAAGTTACATATTGAAAGAGAAGAAATGGTATCATCAAAAAAGC

Cucumis sativus

CAAAGAAATTACTGTTCTTTGCGATGCTCA

Ser22Term
TGAGCATCGCAAAGAACAGTAATTTCTTTGGCTTTTTTGATGATAC
802

TCA-TGA
CATTTCTTCTCTTTCAATATGTAACTTGTCTATTGCTTGAGTTCTCTA

TTGTTTTTATTTCTATTTTCCCTCTTC

TACATATTGAAAGAGAA
803

TTCTCTTT+E,un CAATATGTA
804

Male-sterile
AGAGGGAAAATAGAAATAAAAAGAATAGAGAACTCAAGCAATAGAC
805

P1
AAGTTAGATATTCATAGAGAAGAAATGGTATCATCAAAAAAGCCAA

Cucumis sativus

AGAAATTACTGTTCTTTGCGATGCTCAAG

Lys23Term
CTTGAGCATCGCAAAGAACAGTAATTTCTTTGGCTTTTTTGATGATA
806

AAG-TAG
CCATTTCTTCTCTATGAATATGTAACTTGTCTATTGCTTGAGTTCTC

TATTCTTTTTATTTCTATTTTCCCTCT

CATATTCATAGAGAAGA
807

TCTTCTCTATGAATATG
808

Male-sterile
GGGAAAATAGAAATAAAAAGAATAGAGAACTCAAGCAATAGACAAG
809

P1
TTACATATTCAAAGTGAAGAAATGGTATCATCAAAAAAGCCAAAGA

Cucumis sativus

AATTACTGTTCTTTGCGATGCTCAAGTTT

Arg24Term
AAACTTGAGCATCGCAAAGAACAGTAATTTCTTTGGCTTTTTTGATG
810

AGA-TGA
ATACCATTTCTTCACTTTGAATATGTAACTTGTCTATTGCTTGAGTT

CTCTATTCTTTTTATTTCTATTTTCCC

ATTCAAAGTGAAGAAAT
811

ATTTCTTCACTTTGAAT
812

Male-sterile
GGGACGTGGGAAGGTTGAGATCAAGAGGATTGAGAACTGAAGTAA
813

P1
CAGGCAGGTGACCTAGTCCAAGAGGAGGAATGGGATTATCAAGAA

Malus domestica

GGCAAAGGAGATCACTGTTCTATGTGATGCT

Tyr21Term
AGCATCACATAGAACAGTGATCTCCTTTGCCTTCTTGATAATCCCA
814

TAG-TAG
TTCCTCCTCTTGGACTAGGTGACCTGCCTGTTACTTGAGTTCTCAA

TCCTCTTGATCTCAACCTTCCCACGTCGC

GTGACCTAGTGCAAGAG
815

CTCTTGGACTAGGTCAC
816

Male-sterile
CGTGGGAAGGTTGAGATCAAGAGGATTGAGAACTCAAGTAACAGG
817

P1
CAGGTGACCTACTCCTAGAGGAGGAATGGGATTATCAAGAAGGCA

Malus domestica

AAGGAGATCACTGTTCTATGTGATGCTAAAG

Lys23Term
CTTTAGCATCACATAGAACAGTGATCTCCTTTGCCTTCTTGATAATC
818

AAG-TAG
CCATTCCTCCTCTAGGAGTAGGTCACCTGCCTGTTACTTGAGTTCT

CAATCCTCTTGATCTCAACCTTCCCACG

CCTACTCCTAGAGGAGG
819

CCTCCTCTAGGAGTAGG
820

Male-sterile
AGGATTGAGAAGTCAAGTAACAGGCAGGTGACCTACTCCAAGAGG
821

P1
AGGAATGGGATTATCTAGAAGGCAAAGGAGATGACTGTTCTATGT

Malus domestica

GATGCTAAAGTATCTCTTATCATTTATTCTA

Lys30Term
TAGAATAAATGATAAGAGATACTTTAGCATCACATAGAACAGTGAT
822

AAG-TAG
CTCCTTTGCCTTCTAGATAATCGCATTCCTCCTCTTGGAGTAGGTC

ACCTGCCTGTTACTTGAGTTCTCAATCCT

GGATTATCTAGAAGGCA
823

TGCCTTCTAGATAATCC
824

Male-sterile
ATTGAGAACTCAAGTAACAGGCAGGTGACCTACTCCAAGAGGAGG
825

P1
AATGGGATTATCAAGTAGGCAAAGGAGATCACTGTTCTATGTGATG

Malus domestica

CTAAAGTATCTCTTATCATTTATTCTAGCT

Lys31Term
AGCTAGAATAAATGATAAGAGATACTTTAGCATCACATAGAACAGT
826

AAG-TAG
GATCTCCTTTGCCTACTTGATAATGCCATTCCTCCTCTTGGAGTAG

GTCACCTGCCTGTTACTTGAGTTCTCAAT

TTATCAAGTAGGCAAAG
827

CTTTGCCTACTTGATAA
828

Male-sterile
CATTTTTACAATAGTTATCTGCAAACAAAAACAAGAGAGAAAAACAA
829

globosa
AAACAAAAAAATGTGAAGAGGAAAAATTGAGATCAAAAGAATTGAG

Antirrhinum majus

AACTCAAGCAACAGGCAGGTTACTTACT

Gly2Term
AGTAAGTAACCTGCCTGTTGCTTGAGTTCTCAATTCTTTTGATCTCA
830

GGA-TGA
ATTTTTCCTCTTCACATTTTTTTGTTTTTGTTTTTCTCTCTTGTTTTTG

TTTGCAGATAACTATTGTAAAAATG

AAAAAATGTGAAGAGGA
831

TCCTCTTCACATTTTTT
832

Male-sterile
TTTTACAATAGTTATCTGCAAACAAAAACAAGAGAGAAAAACAAAAA
833

globosa
CAAAAAAATGGGATGAGGAAAAATTGAGATCAAAAGAATTGAGAAC

Antirrhinum majus

TCAAGCAACAGGCAGGTTACTTACTCAA

Arg3Term
TTGAGTAAGTAACCTGCCTGTTGCTTGAGTTCTCAATTCTTTTGATC
834

AGA-TGA
TCAATTTTTCCTCATCCCATTTTTTTGTTTTTGTTTTTCTCTCTTGTTT

TTGTTTGCAGATAACTATTGTAAAA

AAATGGGATGAGGAAAA
835

TTTTCCTCATCCCATTT
836

Male-sterile
TACAATAGTTATCTGCAAACAAAAACAAGAGAGAAAAACAAAAACA
837

globosa
AAAAAATGGGAAGATGAAAAATTGAGATCAAAAGAATTGAGAACTC

Antirthinum majus

AAGCAACAGGCAGGTTACTTACTCAAAGA

Gly4Term
TCTTTGAGTAAGTAACCTGCCTGTTGCTTGAGTTCTCAATTCTTTTG
838

GGA-TGA
ATCTCAATTTTTCATCTTCCCATTTTTTTGTTTTTGTTTTTCTCTCTTG

TTTTTGTTTGCAGATAACTATTGTA

TGGGAAGATGAAAAATT
839

AATTTTTCATCTTCCCA
840

Male-sterile
AATAGTTATCTGCAAACAAAAACAAGAGAGAAAAACAAAAACAAAA
841

globosa
AAATGGGAAGAGGATAAATTGAGATCAAAAGAATTGAGAACTCAAG

Antirrhinum majus

CAACAGGCAGGTTACTTACTCAAAGAGAA

Lys5Term
TTCTCTTTGAGTAAGTAACCTGCCTGTTGCTTGAGTTCTCAATTGTT
842

AAA-TAA
TTGATCTCAATTTATCCTCTTCCCATTTTTTTGTTTTTGTTTTTCTCT

CTTGTTTTTGTTTGCAGATAACTATT

GAAGAGGATAAATTGAG
843

CTCAATTTATCCTCTTC
844

Male-sterile
GCTGAGCTCTTGCTGCCCTTGGATCTGTTTGGGAGTGGAGAACGC
845

P1
AGTATGGGGCGCGGCTAGATCAAGATCAAGAGGATCGAGAACTCT

Zea mays

ACCAACCGGCAGGTGACCTTCTCCAAGCGCC

Lys5Term
GGCGCTTGGAGAAGGTCACCTGCCGGTTGGTAGAGTTCTCGATCC
846

AAG-TAG
TCTTGATCTTGATCTAGCCGCGCCCCATACTGCGTTCTCCACTCCC

AAACAGATCCAAGGGCAGCAAGAGCTCAGC

GGCGCGGCTAGATGAAG
847

CTTGATCTAGCCGCGCC
848

Male-sterile
CTCTTGCTGCCCTTGGATCTGTTTGGGAGTGGAGAACGCAGTATG
849

P1
GGGCGCGGCAAGATCTAGATCAAGAGGATCGAGAACTCTACCAAC

Zea mays

CGGCAGGTGACCTTCTCCAAGCGCCGGGCCG

Lys7Term
CGGCCCGGCGCTTGGAGAAGGTCACCTGCCGGTTGGTAGAGTTC
850

AAG-TAG
TCGATCCTCTTGATCTAGATCTTGCCGCGCCCCATACTGCGTTCTC

CACTCCCAAACAGATCCAAGGGCAGCAAGAG

GCAAGATCTAGATCAAG
851

CTTGATCTAGATCTTGC
852

Male-sterile
CTCTTGCTGCCCTTGGATCTGTTTGGGAGTGGAGAACGCAGTATG
853

P1
GGGCGCGGCAAGATCTAGATCAAGAGGATCGAGAACTCTACCAAC

Zea mays

CGGCAGGTGACCTTCTCCAAGCGCCGGGCCG

Lys9Term
CGGCCCGGCGCTTGGAGAAGGTCACCTGCCGGTTGGTAGAGTTC
854

AAG-TAG
TCGATCCTCTTGATCTAGATCTTGCCGCGCCCCATACTGCGTTCTC

CACTCCCAAACAGATCCAAGGGCAGCAAGAG

GCAAGATCTAGATCAAG
855

GTTGATCTAGATCTTGC
856

Male-sterile
GATCTGTTTGGGAGTGGAGAACGCAGTATGGGGCGCGGCAAGAT
857

P1
CAAGATCAAGAGGATCTAGAACTCTACCAACCGGCAGGTGACCTT

Zea mays

CTCCAAGCGCCGGGCCGGACTGGTCAAGAAGG

Glu12Term
CCTTCTTGACGAGTCCGGCCCGGCGCTTGGAGAAGGTCACCTGC
858

GAG-TAG
CGGTTGGTAGAGTTCTAGATCCTCTTGATCTTGATCTTGCCGCGCC

CCATACTGCGTTCTCCACTCCCAAACAGATC

AGAGGATCTAGAACTCT
859

AGAGTTCTAGATGCTCT
860

Male-sterile
GCTGAGCTCTTGCTGCCCTTGAATCTGTTAGGGAGTGGAGAACGG
861

P1
AGTATGGGGCGCGGCTAGATCGAGATCAAGAGGATCGAGAACTCT

Zea mays

ACCAACCGGCAGGTGACCTTCTCCAAGCGCC

Lys5Term
GGCGCTTGGAGAAGGTCACCTGCCGGTTGGTAGAGTTCTCGATCC
862

AAG-TAG
TCTTGATCTCGATCTAGCCGCGCCCCATACTCCGTTCTCCACTCCC

TAACAGATTCAAGGGCAGCAAGAGCTCAGC

GGCGCGGCTAGATCGAG
863

CTCGATCTAGCCGCGCC
864

Male-sterile
CTCTTGCTGCCCTTGAATCTGTTAGGGAGTGGAGAACGGAGTATG
865

P1
GGGCGCGGCAAGATCTAGATCAAGAGGATCGAGAACTCTACCAAC

Zea mays

CGGCAGGTGACCTTCTCCAAGCGCCGGGCCG

Glu7Term
CGGCCCGGCGCTTGGAGAAGGTCACCTGCCGGTTGGTAGAGTTC
866

GAG-TAG
TCGATCCTCTTGATCTAGATCTTGCCGCGCCCCATACTCCGTTCTC

CACTCCCTAACAGATTCAAGGGCAGCAAGAG

GCAAGATCTAGATCAAG
867

CTTGATCTAGATCTTGC
868

Male-sterile
CTGCCCTTGAATCTGTTAGGGAGTGGAGAACGGAGTATGGGGCG
869

P1
CGGCAAGATCGAGATCTAGAGGATCGAGAACTCTACCAACCGGCA

Zea mays

GGTGACCTTCTCCAAGCGCCGGGCCGGACTGG

Lys9Term
CCAGTCCGGCCCGGCGCTTGGAGAAGGTCACCTGCCGGTTGGTA
870

AAG-TAG
GAGTTCTCGATCCTCTAGATCTCGATCTTGCCGCGCCCCATACTC

CGTTCTCCACTCCCTAACAGATTCAAGGGCAG

TCGAGATCTAGAGGATC
871

GATCCTCTAGATCTCGA
872

Male-sterile
AATCTGTTAGGGAGTGGAGAACGGAGTATGGGGCGCGGCAAGAT
873

P1
GGAGATGAAGAGGATCTAGAACTCTACCAACCGGCAGGTGACCTT

Zea mays

CTCCAAGCGCCGGGCCGGACTGGTCAAGAAGG

Glu12Term
CCTTCTTGACCAGTCCGGCCCGGCGCTTGGAGAAGGTCACCTGC
874

GAG-TAG
CGGTTGGTAGAGTTCTAGATCCTCTTGATCTCGATCTTGCCGCGC

CCCATACTCCGTTCTCCACTCCCTAACAGATT

AGAGGATCTAGAACTCT
875

AGAGTTCTAGATCCTCT
876

Male-sterile
TTGCTGCTAAGCTAGCTGGAGGAAGGAGGAGGAGGAGGAGGAGG
877

P1
CGGGATGGGGCGCGGG+E,un TAGATCGAGATCAAGAGGATCGAGAACT

Oryza sativa

CCACCAACCGCCAGGTGACCTTCTCCAAGCGCA

Lys5Term
TGCGCTTGGAGAAGGTCACCTGGCGGTTGGTGGAGTTCTCGATCC
878

AAG-TAG
TCTTGATGTCGATGTACCCGCGCCCCATCCCGCCTCCTCCTCCTC

CTCCTCCTTCCTCCAGCTAGCTTAGCAGCAA

GGCGCGGGTAGATCGAG
879

CTCGATCTACCCGCGCC
880

Male-sterile
CTAAGCTAGCTGGAGGAAGGAGGAGGAGGAGGAGGAGGCGGGA
881

P1
TGGGGCGCGGGAAGATCTAGATCAAGAGGATCGAGAACTCCACC

Oryza sativa

AACCGCCAGGTGACCTTCTCCAAGCGCAGGAGCG

Glu7Term
CGCTCCTGCGCTTGGAGAAGGTCACCTGGCGGTTGGTGGAGTTCT
882

GAG-TAG
CGATCCTCTTGATCTAGATCTTCCCGCGCCCCATCCCGCCTCCTC

CTCCTCCTCCTCCTTCCTCCAGCTAGCTTAG

GGAAGATCTAGATCAAG
883

CTTGATCTAGATCTTCC
884

Male-sterile
TAGCTGGAGGAAGGAGGAGGAGGAGGAGGAGGCGGGATGGGGC
885

P1
GCGGGAAGATCGAGATCTAGAGGATCGAGAACTCCACCAACCGC

Oryza sativa

CAGGTGACCTTCTCCAAGCGCAGGAGCGGGATCC

Lys9Term
GGATCCCGCTCCTGCGCTTGGAGAAGGTCACCTGGCGGTTGGTG
886

AAG-TAG
GAGTTCTCGATCCTCTAGATCTCGATCTTCCCGCGCCCCATCCCG

CCTCCTCCTCCTCCTCCTCCTTCCTCCAGCTA

TCGAGATCTAGAGGATC
887

GATCCTCTAGATCTCGA
888

Male-sterile
GAAGGAGGAGGAGGAGGAGGAGGCGGGATGGGGCGCGGGAAG
889

P1
ATCGAGATCAAGAGGATCTAGAACTCCACCAACCGCCAGGTGACC

Oryza sativa

TTCTCCAAGCGCAGGAGCGGGATCCTCAAGAAGG

Glu12Term
CCTTCTTGAGGATCCCGCTCCTGCGCTTGGAGAAGGTCACCTGGC
890

GAG-TAG
GGTTGGTGGAGTTCTAGATCCTCTTGATCTCGATCTTCCCGCGCC

CCATCCCGCCTCCTCCTCCTCCTCCTCCTTC

AGAGGATCTAGAACTCC
891

GGAGTTCTAGATCCTCT
892

EXAMPLE 7

Engineering Plants for Abiotic Stress Tolerance

[0128] Environmental stresses, such as drought, increased soil salinity, soil contamination with heavy meals, and extreme temperature, are major factors limiting plant growth and productivity. The worldwide loss in yield of three major cereal crops, rice, maize, and wheat due to water stress (drought) has been estimated to be over ten billion dollars annually and many currently marginal soils could be brought into cultivation if suitable plant varieties were available.

[0129] Physiological and biochemical responses to high levels of ionic or nonionic solutes and decreased water potential have been studied in a variety of plants. It is known, for example, that increasing levels of alcohol dehydrogenase can confer enhances flooding resistance in plants. There are also several possible mechanisms to enhance plant salt tolerance. For example, one mechanism underlying the adaptation or tolerance of plants to osmotic stresses is the accumulation of compatible, low molecular weight osmolytes such as sugar alcohols, special amino acids, and glycinebetaine. Such accumulation can be engineered, for example, by removing feedback inhibition on 1-pyrroline-t-carboxylate synthetase, which results in accumulation of proline. Additionally, recent experiments suggest that altering the expression or activity of specific sodium or potassium transporters can confer enhanced salt tolerance.

[0130] Plant tolerance of contamination by heavy metals such as lead and aluminum in soils has also been investigated and one mechanism underlying tolerance is the production of dicarboxylic acids such as oxalate and citrate. In addition, individual genes involved in heavy metal sensitivity have been identified.

[0131] The attached tables disclose exemplary oligonucleotide base sequences which can be used to generate site-specific mutations that confer stress tolerance in plants.

19TABLE 17Genome-Altering Oligos Conferring Stress TolerancePhenotype, Gene,Plant & TargetedSEQ IDAlterationAltering OligosNO:Salt ToleranceCGTCTTTTTGTGTGGTAGTTGGATGTGACGGTTGCTCAAATGCTT893P5CSGTGACCGATAGCAGTGCTAGAGATAAGGATTTCAGGAAGCAACTTArabidopsis thalianaAGTGAAACTGTCAAAGCGATGCTGAGGATGAPhe128AlaTCATCCTCAGCATCGCTTTGACAGTTTCACTAAGTTGCTTCCTGAA894TTT-GCTATCCTTATGTCTAGCACTGCTATCGGTCACAAGCATTTGAGCAACCGTCACATCCAACTACCACACAAAAAGACGATAGCAGTGCTAGAGAT895ATCTCTAGCACTGCTAT896Salt ToleranceGAGAGTATGTTTGACCAGCTGGATGTGACGGCTGCTCAGCTGCTG897P5CS 1GTGAATGACAGTAGTGCCAGAGACAAGGAGTTCAGGAAGCAACTTBrassica napusAATGAGACAGTGAAGTCCATGCTTGATTTGAPhe128AlaTCAAATCAAGCATGGACTTCACTGTCTCATTAAGTTGCTTCCTGAA898TTC-GCCCTCCTTGTCTCTGGCACTACTGTCATTCACCAGCAGCTGAGCAGCCGTCACATCCAGCTGGTCAAACATAGTGTCACAGTAGTGCCAGAGAC899GTCTCTGGCACTACTGT900Salt ToleranceGAGACTATGTTTGACCAGATGGATGTGACGGTGGCTCAAATGCTG901P505 2GTGACTGATAGCAGTGTCAGAGATAAGGATTTCAGGAAGCAACTTBrassica napusAGTGAGACAGTCAAAGCTATGCTGAAAATGAPhe129AlaTCATTTTCAGCATAGCTTTGACTGTCTCACTAAGTTGCTTCCTGAA902TTC-GCCATCCTTATCTCTGACACTGCTATCAGTCACCAGCATTTGAGCCACCGTCACATCCATCTGGTCAAACATAGTCTCATAGCAGTGTCAGAGAT903ATCTCTGACACTGCTAT904Salt ToleranceGATATGTTGTTTAACCAACTGGATGTCTCGTCATCTCAACTTCTTG905P5GSTCACCGACAGTGATGCTGAGAACCCAAAGTTCCGGGAGCAACTCAOryza sativaCTGAAACTGTTGAGTCATTATTAGATCTTAPhe128AlaTAAGATCTAATAATGACTCAACAGTTTCAGTGAGTTGCTCCCGGAA906TTT-GCTCTTTGGGTTCTCAGCATCACTGTCGGTGACAAGAAGTTGAGATGACGAGACATCCAGTTGGTTAAACAACATATCACAGTGATGCTGAGAAC907GTTCTCAGCATCACTGT908Salt ToleranceGATATTTTGTTTAGTCAGCTGGATGTGACATCTGCTCAGCTTCTTG909P5CSTTACTGACAATGATGCTAGAGACCAAGATTTTAGAAAGCAACTTTCMedicago sativaTGAAACTGTGAGATCACTTCTAGCACTAAPhe128AlaTTAGTGCTAGAAGTGATCTCACAGTTTCAGAAAGTTGCTTTCTAAA910TTT-GCTATCTTGGTCTCTAGCATCATTGTCAGTAAGAAGAAGCTGAGCAGATGTCACATCCAGCTGACTAAACAAAATATCACAATGATGCTAGAGAC911GTCTCTAGCATCATTGT912Salt ToleranceGATACATTGTTTAGTCAGCTGGATGTGACATCAGCTCAGCTACTC913P5CSGTTACTGATAATGATGCTAGGGATCCAGAATTCAGGAAGCAACTTActinidia deliciosaACTGAAACTGTAGAATCACTATTGAATTTGAPhe128AlaTCAAATTCAATAGTGATTCTACAGTTTCAGTAAGTTGCTTCCTGAAT914TTT-GCTTCTGGATCCCTAGCATCATTATCAGTAACGAGTAGCTGAGCTGATGTCACATCCAGCTGACTAAACAATGTATCATAATGATGCTAGGGAT915ATCCCTAGCATCATTAT916Salt ToleranceGACACACTCTTCAGTCAACTGGATGTGACATCAGCACAGCTTCTT917P5CSGTAACAGATAATGACGCCAGAAGTCCAGAATTTAGAAAACAACTTACichorium intybusCTGAAACAGTCGATTCTTTATTATCTTATAPhe122AlaTATAAGATAATAAAGAATCGACTGTTTCAGTAAGTTGTTTTCTAAAT918TTC-GCCTCTGGACTTCTGGCGTCATTATCTGTTACAAGAAGCTGTGCTGATGTCACATCCAGTTGACTGAAGAGTGTGTCATAATGACGCCAGAAGT919ACTTCTGGCGTCATTAT920Salt ToleranceGATTCTTTGTTCAGTCAGTTGGATGTGACATCAGCTCAGCTTCTGG921P5CSTGACTGATAATGACGCTAGAGATCCAGATTTTAGGAGACAACTCALycopersiconATGACACAGTAAATTCGTTGCTTTCTCTAAesculentumTTAGAGAAAGCAACGAATTTACTGTGTCATTGAGTTGTCTCCTAAA922Phe12BAlaATCTGGATCTCTAGCGTCATTATCAGTCACCAGAAGCTGAGCTGATTT-GCTTGTCACATCCAACTGACTGAACAAAGAATCATAATGACGCTAGAGAT923ATCTCTAGCGTCATTAT924Salt ToleranceGATACCATGTTCAGCCAGCTTGATGTGACTTCTTCCCAACTTCTTG925P5CSTGAATGATGGATTTGCTAGGGATGCTGGCTTCAGAAAACAACTTTVigna unguiculataCGGACACAGTGAACGCGTTATTAGATTTAAPhe162AlaTTAAATCTAATAACGCGTTCACTGTGTCCGAAAGTTGTTTTCTGAA926TTT-GCTGCCAGCATCCCTAGCAAATCCATCATTCACAAGAAGTTGGGAAGAAGTCACATCAAGCTGGCTGAACATGGTATCATGGATTTGCTAGGGAT927ATCCCTAGCAAATCCAT928 Salt ToleranceGACACCTTGTTTAGTCAGTTGGATCTGACTGCTGCTCAGCTGCTT929P5CSGTGACGGACAACGACGCTAGAGATCCAAGTTTTAGAACACAACTAMesembryanthemumACTGAAACAGTGTATCAGTTGTTGGATCTAAcrystallinumTTAGATCCAACAACTGATACACTGTTTCAGTTAGTTGTGTTCTAAA930Phe125AlaACTTGGATCTCTAGCGTCGTTGTCCGTCACAAGCAGCTGAGCAGCTTT-GCTAGTCAGATCCAACTGACTAAACAAGGTGTCACAACGACGCTAGAGAT931ATCTCTAGCGTCGTTGT932Salt ToleranceGACACATTATTTAGCCAGCTGGATGTGACATCAGCTCAGCTTCTT933P5CSGTGACTGATAATGATGCTAGGGATGAAGCTTTCCGAAATCAACTTAVitis viniferaCTCAAACAGTGGATTCATTGTTAGCTTTGAPhe130AlaTCAAAGCTAACAATGAATCCACTGTTTGAGTAAGTTGATTTCGGAA934TTT-GCTAGCTTCATCCCTAGCATCATTATCAGTCACAAGAAGCTGAGCTGATGTCACATCCAGCTGGCTAAATAATGTGTCATAATGATGCTAGGGAT935ATCCCTAGCATCATTAT936Salt ToleranceGATACGCTGTTCACTCAGCTCGATGTGACATCGGCTCAGCTTCTT937P5CSGTGACGGATAACGATGCTCGAGATAAGGATTTCAGGAAGCAGCTTVigna aconitifoliaACTGAGACTGTGAAGTCGCTGTTGGGGCTGAPhe129AlaTCAGCGCCAACAGCGACTTCACAGTCTCAGTAAGCTGCTTCCTGA938TTT-GCTAATCCTTATCTCGAGCATCGTTATCCGTCACAAGAAGCTGAGCCGATGTCACATCGAGCTGAGTGAACAGCGTATCATAACGATGCTCGAGAT939ATCTCGAGCATCGTTAT940Salt ToleranceAGAGATGTTCTTAGTTCCAAAGAAATCTCACCTCTCAGTTTCTCCG941HKT1TCTTCACAACAGTTGTCACGTTTGCAAACTGCGGATTTGTCCCCACArabidopsis thalianaGAATGAGAACATGATCATCTTTCGCAAAASer207ValTTTTGCGAAAGATGATCATGTTCTCATTCGTGGGGACAAATCCGC942TCC-GTCAGTTTGCAAACGTGACAACTGTTGTGAAGACGGAGAAAGTGAGAGGTGAGATTTCTTTGGAACTAAGAACATCTCTCAACAGTTGTCACGTTT943AAACGTGACAACTGTTG944Salt ToleranceCGAATGAGAACATGATCATCTTTCGCAAAAACTCTGGTCTCATCTG945HKT1GCTCCTAATCCCTCTAGTACTGATGGGAAACACTTTGTTCCCTTGCArabidopsis thalianaTTCTTGGTTTTGCTCATATGGGGACTTTAGln237LeuTAAAGTCCCCATATGAGCAAAACCAAGAAGCAAGGGAACAAAGTG946CAA-CTATTTCCCATCAGTACTAGAGGGATTAGGAGCCAGATGAGACCAGAGTTTTTGCGAAAGATGATCATGTTCTCATTCGAATCCCTCTAGTACTGA947TCAGTACTAGAGGGATT948Salt ToleranceAGTCTCTAGAAGGAATGAGTTCGTACGAGAAGTTGGTTGGATCGT949HKT1TGTTTCAAGTGGTGAGTTCGCGACACACCGGAGAAACTATAGTAGArabidopsis thalianaACCTCTCTACACTTTCCCCAGCTATCTTGGTAsn332SerACCAAGATAGCTGGGGAAAGTGTAGAGAGGTCTACTATAGTTTCT950AAT-AGTCCGGTGTGTCGCGAACTCACCACTTGAAACAACGATCCAACCAACTTCTCGTACGAACTCATTCCTTCTAGAGACTAGTGGTGAGTTCGCGAC951GTCGCGAACTCACCACT952Salt ToleranceAGAGATGTGCTAAAGAAGAAAGGTCTCAAAATGGTGACCTTTTCC953HKT1GTCTTCACCACCGTGGTGACCTTTGCCAGTTGTGGGTTTGTCCCGEucalyptusACCAATGAAAACATGATTATCTTCAGCAAAAcamaldulensisTTTTGCTGAAGATAATCATGTTTTCATTGGTCGGGACAAACCCACA954Ser256ValACTGGCAAAGGTCACCACGGTGGTGAAGACGGAAAAGGTCACCATCG-GTGTTTTGAGACCTTTCTTCTTTAGCACATCTCTCCACCGTGGTGACCTTT955AAAGGTCACCACGGTGG956Salt ToleranceCCAATGAAAACATGATTATCTTCAGCAAAAACTCTGGCCTCCTCCT957HKT1GATTCTCATCCCTCTGGCCCTTCTTGGGAACATGCTGTTCCCATCEucalyptusGAGCCTACGTTTGACGCTTTGGCTCATCGGcamaldulensisCCGATGAGCCAAAGCGTCAAACGTAGGCTCGATGGGAACAGCAT958Gln286LeuGTTCCCAAGAAGGGCCAGAGGGATGAGAATCAGGAGGAGGCCACAG-CTGGAGTTTTTGCTGAAGATAATCATGTTTTCATTGGCATCCCTCTGGCCCTTC959GAAGGGCCAGAGGGATG960Salt ToleranceAATCGTTGAATGGACTAAGCTCCTGTGAGAAAATCGTGGGCGCGC961HKT1TGTTTCAGTGCGTGAGCAGCAGACATACCGGCGAGACGGTCGTCEucalyptusGATCTGTCCACAGTTGCTCCCGCCATCTTGGTcamaldulensisACCAAGATGGCGGGAGCAACTGTGGACAGATCGACGACCGTCTC962Asn381SerGCCGGTATGTCTGCTGCTCACGCACTGAAACAGCGCGCCCACGAAAC-AGCTTTTCTCACAGGAGCTTAGTCCATTCAACGATTGTGCGTGAGCAGCAGAC963GTCTGCTG+E,un CTCACGCAC964Salt ToleranceAAAGCTCCACTGAAGAAGAAAGGGATCAACATTGCACTCTTCTCA965HKT1TTCTCGGTCACGGTCGTCTCGTTTGCGAATGTGGGGCTCGTGCCOryza sativaGACAAATGAGAACATGGCAATCTTCTCCAAGASer238ValTCTTGGAGAAGATTGCCATGTTCTCATTTGTCGGCACGAGCCCCA966TCC-GTCCATTCGCAAACGAGACGACCGTGACCGAGAATGAGAAGAGTGCAATGTTGATCCCTTTCTTCTTCAGTGGAGCTTTTCACGGTCGTCTCGTTT967AAACGAGACGACCGTGA968Salt ToleranceCAAATGAGAACATGGCAATCTTCTCCAAGAACCCGGGCCTCCTCC969HKT1TCCTGTTCATCGGCCTGATTGTTGCAGGCAATACACTTTACCCTCTOryza sativaCTTCCTAAGGCTATTGATATGGTTCCTGGGGln268LeuCCCAGGAACCATATCAATAGCCTTAGGAAGAGAGGGTAAAGTGTA970CAG-CTGTTGCCTGCAAGAATCAGGCCGATGAACAGGAGGAGGAGGCCCGGGTTCTTGGAGAAGATTGCCATGTTCTCATTTGCATCGGCCTGATTCTTG971CAAGAATCAGGCCGATG972Salt ToleranceCAGTCTTTGATGGACTCAGCTCTTACCAGAAGATTATCAATGCATT973HKT1GTTCATGGCAGTGAGCGCAAGGCACTCGGGGGAGAACTCCATCGOryza sativaACTGCTCACTCATCGCCCCTGCTGTTCTAGTAsn363SerACTAGAACAGCAGGGGCGATGAGTGAGCAGTCGATGGAGTTCTC974AAC-AGCCCCCGAGTGCCTTGCGCTCACTGCCATGAACAATGCATTGATAATCTTCTGGTAAGAGCTGAGTCCATCAAAGACTGGGCAGTGAGCGCAAGGC975GCCTTGCGCTCACTGCC976Salt ToleranceGTGCCCCACTGAACAAGAAAGGGATCAACATCGTGCTCTTCTCAC977HKT1TATCAGTCACCGTTGTCTCCTGTGCGAATGCAGGACTCGTGCCCATriticum aestivumCAAATGAGAACATGGTCATCTTCTCAAAGAAAla240ValTTCTTTGAGAAGATGACCATGTTCTCATTTGTGGGCACGAGTCCT978GCC-GTCGCATTCGCACAGGAGACAACGGTGAGTGATAGTGAGAAGAGCACGATGTTGATCCCTTTCTTGTTCAGTGGGGCACCACCGTTGTCTCCTGTG979CACAGGAGACAACGGTG980Salt ToleranceCAAATGAGAACATGGTCATCTTCTCAAAGAATTCAGGCCTCTTGTT981HKT1GCTGCTGAGTGGCCTGATGCTCGCAGGCAATACATTGTTCCCTCTTriticum aestivumCTTCCTGAGGCTACTGGTGTGGTTCCTGGGGln270LeuCCCAGGAACCACACCAGTAGCCTCAGGAAGAGAGGGAACAATGT982CAG-CTGATTGCCTGCGAGCATCAGGCCACTCAGCAGCAACAAGAGGCCTGAATTCTTTGAGAAGATGACCATGTTCTCATTTGGAGTGGCCTGATGCTCG983CGAGCATCAGGCCACTC984Salt ToleranceCAGTCTTTGATGGGCTCAGCTCTTATCAGAAGACTGTCAATGCATT985HKT1CTTCATGGTGGTGAGTGCGAGGCACTCAGGGGAGAATTCCATCGTriticum aestivumACTGCTCGCTCATGTCCCCTGCCATTATAGTAsn365SerACTATAATGGCAGGGGACATGAGCGAGCAGTCGATGGAATTCTCC986AAT-AGTCCTGAGTGCCTCGCACTCACCACCATGAAGAATGCATTGACAGTCTTCTGATAAGAGCTGAGCCCATCAAAGACTGGGTGGTGAGTGCGAGGC987GCCTCGCACTCACCACC988Freezing ToleranceTTTTTTTTGTTTTCGTTTTCAAAAAGAAAATCTTTGAATTTTATGGCA989praline oxidaseACCGGTCTTCTCTGAACAAACTTTATCCGGCGATCTTACCGTTTAGprecursorCCGCTTTTAGCCCGGTGGGTCCTCCCAArabidopsis thalianaTGGGAGGACCCACCGGGCTAAAAGCGGGTAAACGGTAAGATCGC990Arg7TermGGGATAAAGTTTGTTCAGAGAAGACGGGTTGCCATAAAATTCAAACGA-TGAGATTTTGTTTTTGAAAACGAAAACAAAAAAAAGTCTTCTCTGAACAAAC991GTTTGTTCAGAGAAGAC992Freezing ToleranceTCAAAAACAAAATCTTTGAATTTTATGGCAACCCGTCTTCTCAGAA993proline oxidaseCAAACTTTATCCGGTGATCTTACCGTTTACCGGCTTTTAGCCCGGTprecursorGGGTCCTCCCACCGTGACTGCTTCCACCGArabidopsis thalianaCGGTGGAAGCAGTCACGGTGGGAGGACCCAGCGGGCTAAAAGC994Arg13TermGGGTAAACGGTAAGATCACCGGATAAAGTTTGTTCTGAGAAGACGCGA-TGAGGTTGCCATAAAATTCAAAGATTTTGTTTTTGATTATCCGGTGATCTTAC995GTAAGATCACCGGATAA996Freezing ToleranceAAAATCTTTGAATTTTATGGCAACCCGTCTTCTCCGAACAAACTTT997praline oxidaseATCCGGCGATCTTAGCGTTTACCCGCTTTTAGCCCGGTGGGTCCTprecursorCCCACCGTGACTGCTTCCACCGCCGTCGTCArabidopsis thalianaGACGACGGCGGTGGAAGCAGTCACGGTGGGAGGACCCACCGGG998Tyr15TermCTAAAAGCGGGTAAACGCTAAGATCGCCGGATAAAGTTTGTTCGGTAG-TAGAGAAGAGGGGTTGCCATAAAATTCAAAGATTTTCGATCTTAGCGTTTACC999GGTAAACGCTAAGATCG1000Freezing ToleranceCTTTGAATTTTATGGCAACCCGTCTTCTCCGAACAAACTTTATCCG1001praline oxidaseGCGATCTTACCGTTAACCCGCTTTTAGCCCGGTGGGTCCTCCCACprecursorCGTGACTGCTTCCACCGCCGTCGTCCCGGAArabidopsis thalianaTCCGGGACGACGGCGGTGGAAGCAGTCACGGTGGGAGGACCCA1002Leu17TermCCGGGCTAAAAGCGGGTTAACGGTAAGATCGGCGGATAAAGTTTTTA-TAAGTTCGGAGAAGACGGGTTGCCATAAAATTCAAAGTTACCGTTAACCCGCTT1003AAGCGGGTTAACGGTAA1004Freezing ToleranceCCGGTGGGTCCTCCCACCGTGACTGCTTCCAGCGCCGTGGTCCC1005proline oxidaseGGAGATTCTCTCCTTTTGACAACAAGCACCGGAACCACCTCTTCAprecursorCCACCCAAAACCCACCGAGCAATCTCACGATGArabidopsis thalianaCATCGTGAGATTGCTCGGTGGGTTTTGGGTGGTGAAGAGGTGGT1006Gly42TermTCCGGTGCTTGTTGTCAAAAGGAGAGAATCTCCGGGACGACGGCGGA-TGAGGTGGAAGCAGTCACGGTGGGAGGACCCACCGGTCTCCTTTTGACAACAA1007TTGTTGTCAAAAGGAGA1008Lead ToleranceACATGAAGCAGTGAAATCTCTGTTTGTATTGAATCTTATTAGTCTCT1009cyclic nucleotide-AAACTATGAATTTCTGACAAGAGAAGTTTGTAAGGTCAGTGTTCCAregulated ion channelGATTTGTCTCATTGAATTCTAAGTCGTGAArabidopsis thalianaTCACGACTTAGAATTCAATGAGACAAATCTGGAACACTGACCTTAC1010Arg4TermAAACTTCTCTTGTCAGAAATTCATAGTTTGAGACTAATAAGATTCAACGA-TGATACAAACAGAGATTTCACTGCTTCATGTTGAATTTCTGACAAGAG1011CTCTTGTCAGAAATTCA1012Lead ToleranceTGAAGCAGTGAAATCTCTGTTTGTATTGAATCTTATTAGTCTCAAA1013cyclic nucleotide-CTATGAATTTCCGATAAGAGAAGTTTGTAAGGTCAGTGTTCCAGATregulated ion channelTTGTCTCATTGAATTCTAAGTCGTGAAGCArabidopsis thalianaGCTTCACGACTTAGAATTCAATGAGACAAATCTGGAACACTGACCT1014Gln5TermTACAAACTTCTCTTATCGGAAATTCATAGTTTGAGACTAATAAGATTCAA-TAACAATACAAACAGAGATTTCACTGCTTCAATTTCCGATAAGAGAAG1015CTTCTCTTATCGGAAAT1016Lead ToleranceAGCAGTGAAATCTCTGTTTGTATTGAATCTTATTAGTCTCAAACTAT1017cyclic nucleotide-GAATTTCCGACAATAGAAGTTTGTAAGGTCAGTGTTCCAGATTTGTregulated ion channelCTCATTGAATTCTAAGTCGTGAAGCTTAArabidopsis thalianaTAAGCTTCACGACTTAGAATTCAATGAGACAAATCTGGAACACTGA1018Glu6TermCCTTACAAACTTCTATTGTCGGAAATTCATAGTTTGAGACTAATAAGAG-TAGGATTCAATACAAACAGAGATTTCACTGCTTCCGACAATAGAAGTTT1019AAACTTCTATTGTCGGA1020Lead ToleranceAGTGAAATCTCTGTTTGTATTGAATCTTATTAGTCTCAAACTATGAA1021cyclic nucleotide-TTTCCGACAAGAGTAGTTTGTAAGGTCAGTGTTCCAGATTTGTCTCregulated ion channelATTGAATTCTAAGTCGTGAAGCTTAATTArabidopsis thalianaAATTAAGCTTCACGACTTAGAATTCAATGAGACAAATCTGGAACAC1022Lys7TermTGACCTTACAAACTACTCTTGTCGGAAATTCATAGTTTGAGACTAAAAG-TAGTAAGATTCAATACAAACAGAGATTTCACTGACAAGAGTAGTTTGTA1023TACAAACTACTCTTGTC1024Lead ToleranceCATTGAATTCTAAGTCGTGAAGCTTAATTCGATTCTTCTTCACTTTC1025cyclic nucleotide-TCGGATCAGGTTTTAAGATTGGAAGTCGGATAAGACTTCCTCCGAregulated ion channelCGTGGAATATTCCGGTAAAAACGAGATTCArabidopsis thalianaGAATCTCGTTTTTACCGGAATATTCCACGTCGGAGGAAGTCTTATC1026Gln12TermCGACTTCCAATCTTAAAACCTGATCCGAGAAAGTGAAGAAGAATCCAA-TAAGAATTAAGCTTCACGACTTAGAATTCAATGTCAGGTTTTAAGATTGG1027CCAATCTTAAAACCTGA1028Lead ToleranceTGGAAGTCAATCCCCCACGTTGAGCAGGTTGATGCATTGGGTAAA1029cyclic nucleotide-GTTATGAATCACCGCTAAGACGAGTTTGTGAGGTTTCAGGATTGGgated calmodulin-AAATCAGAGAGAAGCTCTGAGGGAAATTTTCbinding ion channelGAAAATTTCCCTCAGAGCTTCTCTCTGATTTCCAATCCTGAAACCT1030(CBP4)CACAAACTCGTCTTAGCGGTGATTCATAACTTTAGCCAATGCATCANicotiana TabacumACCTGCTCAACGTGGGGGATTGACTTCCAGln5TermATCACCGCTAAGACGAG1031CAA-TAACTCGTCTTAGCGGTGAT1032Lead ToleranceTCAATCCCCCACGTTGAGCAGGTTGATGCATTGGCTAAAGTTATG1033cyclic nucleotide-AATCACCGCCAAGACTAGTTTGTGAGGTTTCAGGATTGGAAATCAgated calmodulin-GAGAGAAGCTCTGAGGGAAATTTTCATGCTAbinding ion channelTAGCATGAAAATTTCCCTCAGAGCTTCTCTCTGATTTCCAATCCTG1034(CBP4)AAACCTCACAAACTAGTCTTGGCGGTGATTCATAACTTTAGCCAATNicotiana TabacumGCATCAACCTGCTCAACGTGGGGGATTGAGly7TermGCCAAGACTAGTTTGTG1035GAG-TAGCACAAACTAGTCTTGGC1036Lead ToleranceGAGCAGGTTGATGCATTGGCTAAAGTTATGAATCACCGCCAAGAC1037cyclic nucleotide-GAGTTTGTGAGGTTTTAGGATTGGAAATCAGAGAGAAGCTCTGAGgated calmodulin-GGAAATTTTCATGCTAAAGGTGGAGTCCACCbinding ion channelGGTGGACTCCACCTTTAGCATGAAAATTTCCCTCAGAGCTTCTCTC1038(CBP4)TGATTTCCAATCCTAAAACCTCACAAACTCGTCTTGGCGGTGATTCNicotiana TabacumATAACTTTAGCCAATGCATCAACCTGCTCGln12TermTGAGGTTTTAGGATTGG1039CAG-TAGCCAATCCTAAAACCTCA1040Lead ToleranceTGATGCATTGGCTAAAGTTATGAATCACCGCCAAGACGAGTTTGT1041cyclic nucleotide-GAGGTTTCAGGATTGTAAATCAGAGAGAAGCTCTGAGGGAAATTTgated calmodulin-TCATGCTAAAGGTGGAGTCCACCGAAGTAAAbinding ion channelTTTACTTCGGTGGACTCCACCTTTAGCATGAAAATTTCCCTCAGAG1042(CBP4)CTTCTCTCTGATTTACAATCCTGAAACCTCACAAACTCGTCTTGGCNicotiana TabacumGGTGATTCATAACTTTAGCCAATGCATCATrp14TermCAGGATTGTAAATCAGA1043TGG-TGATCTGATTTACAATCCTG1044Lead ToleranceGATGCATTGGCTAAAGTTATGAATCACCGCCAAGACGAGTTTGTG1045cyclic nucleotide-AGGTTTCAGGATTGGTAATCAGAGAGAAGCTGTGAGGGAAATTTTgated calmoduin-CATGCTAAAGGTGGAGTCCACCGAAGTAAAGbinding ion channelCTTTACTTCGGTGGACTCCACCTTTAGCATGAAAATTTCCCTCAGA1046(CBP4)GCTTCTCTCTGATTACCAATCCTGAAACCTCACAAACTCGTCTTGGNicotiana TabacumCGGTGATTCATAACTTTAGCCAATGCATCLys15TermAGGATTGGTAATCAGAG1047AAA-TAACTCTGATTACCAATCCT1048Lead ToleranceCTTGAAGAATTGATCTACCACTCTTAGCTGCTAACTGTTCGCCTGG1049calmoduin bindingTGGAGATAATGATGTAAAGAGAGGACAGATATGTTAGATTTCAGGtransport proteinACTGCAAATCAGAGCAATCTGTTATCTCAGHordeum vulgareCTGAGATAACAGATTGCTCTGATTTGCAGTCCTGAAATGTAACATA1050Glu2TermTCTGTCCTCTCTTTACATCATTATCTCCACCAGGCGAACAGTTAGCGAA-TAAAGCTAAGAGTGGTAGATCAATTCTTCAAGTAATGATGTAAAGAGAG1051CTCTCTTTACATCATTA1052Lead ToleranceGAAGAATTGATCTACCACTCTTAGCTGCTAACTGTTCGCCTGGTG1053calmodulin bindingGAGATAATGATGGAATGAGAGGACAGATATGTTAGATTTCAGGACtransport proteinTGCAAATCAGAGCAATCTGTTATCTCAGAGAHordeum vulgareTCTCTGAGATAACAGATTGCTCTGATTTGCAGTCCTGAAATCTAAC1054Arg3TermATATCTGTCCTCTCATTCCATCATTATCTCCACCAGGCGAACAGTTAGA-TGAAGCAGCTAAGAGTGGTAGATCAATTCTTCTGATGGAATGAGAGGAC1055GTCCTCTCATTCCATCA1056Lead ToleranceGAATTGATCTACCACTCTTAGCTGCTAACTGTTCGCCTGGTGGAG1057calmodulin bindingATAATGATGGAAAGATAGGACAGATATGTTAGATTTCAGGACTGCtransport proteinAAATCAGAGCAATCTGTTATCTCAGAGAACGHordeum vulgareCGTTCTCTGAGATAACAGATTGCTCTGATTTGCAGTCCTGAAATCT1058Glu4TermAACATATCTGTCCTATCTTTCCATCATTATCTCCACCAGGCGAACAGAG-TAGGTTAGCAGCTAAGAGTGGTAGATCAATTCTGGAAAGATAGGACAGA1059TCTGTCCTATCTTTCCA1060Lead ToleranceATCTACCACTCTTAGCTGCTAACTGTTCGCCTGGTGGAGATAATG1061calmodulin bindingATGGAAAGAGAGGACTGATATGTTAGATTTCAGGACTGCAAATCAtransport proteinGAGCAATCTGTTATCTCAGAGAACGCAGTTTHordeum vulgareAAACTGCGTTCTCTGAGATAACAGATTGCTCTGATTTGCAGTCCTG1062Arg6TermAAATCTAACATATCAGTCCTCTCTTTCCATCATTATCTCCACCAGGAGA-TGACGAACAGTTAGCAGCTAAGAGTGGTAGATGAGAGGACTGATATGTT1063AACATATCAGTCCTCTC1064Lead ToleranceCCACTCTTAGCTGCTAACTGTTCGCCTGGTGGAGATAATGATGGA1065calmodulin bindingAAGAGAGGACAGATAGGTTAGATTTCAGGAGTGCAAATCAGAGCAtransport proteinATCTGTTATCTCAGAGAACGCAGTTTCACCAHordeum vulgareTGGTGAAACTGCGTTCTCTGAGATAACAGATTGCTCTGATTTGCA1066Tyr7TermGTCCTGAAATCTAACCTATCTGTCCTCTCTTTCCATCATTATCTCCATAT-TAGCCAGGCGAACAGTTAGCAGCTAAGAGTGGGACAGATAGGTTAGATT1067AATCTAACCTATCTGTC10682,4-DB resistanceATCCTTCTCTGAGAAAAAACAACAGATCCGAATTTTATCTTTAATCA10693-ketoacyl-CoAGCCGGAAAAAATGTAGAAAGCGATCGAGAGACAACGCGTTCTTCTthiolaseTGAGCATCTCCGACCTTCTTCTTCTTCTTArabidopsis thalianaAAGAAGAAGAAGAAGGTCGGAGATGCTCAAGAAGAACGCGTTGT1070Glu2TermCTCTCGATCGCTTTCTACATTTTTTCCGGCTGATTAAAGATAAAATTGAG-TAGCGGATCTGTTGTTTTTTCTCAGAGAAGGATAAAAAATGTAGAAAGCG1071CGCTTTCTACATTTTTT10722,4-DB resistanceCTTCTCTGAGAAAAAACAACAGATCCGAATTTTATCTTTAATCAGC10733-ketoacyl-CoACGGAAAAAATGGAGTAAGCGATCGAGAGACAACGCGTTCTTCTTGthiolaseAGCATCTCCGACCTTCTTCTTCTTCTTCGCArabidopsis thalianaGCGAAGAAGAAGAAGAAGGTCGGAGATGCTCAAGAAGAACGCGT1074Lys3TermTGTCTCTCGATCGCTTACTCCATTTTTTCCGGCTGATTAAAGATAAAAA-TAAAATTCGGATCTGTTGTTTTTTCTCAGAGAAGAAATGGAGTAAGCGATC1075GATCGCTTACTCCATTT10762,4-DB resistanceGAAAAAACAACAGATCCGAATTTTATCTTTAATCAGCCGGAAAAAA10773-ketoacyl-CoATGGAGAAAGCGATCTAGAGACAACGCGTTCTTCTTGAGCATCTCCthiolaseGACCTTCTTCTTCTTCTTCGCACAATTACGArabidopsis thalianaCGTAATTGTGCGAAGAAGAAGAAGAAGGTCGGAGATGCTCAAGA1078Glu6TermAGAACGCGTTGTCTCTAGATCGCTTTCTCCATTTTTTCCGGCTGATGAG-TAGTAAAGATAAAATTCGGATCTGTTGTTTTTTCAAGCGATCTAGAGACAA1079TTGTCTCTAGATCGCTT10802,4-DB resistanceAAAACAACAGATCCGAATTTTATCTTTAATCAGCCGGAAAAAATGG10813-ketoacyl-CoAAGAAAGCGATCGAGTGACAACGCGTTCTTCTTGAGCATCTCCGACthiolaseCTTCTTCTTCTTCTTCGCACAATTACGAGGArabidopsis thalianaCCTCGTAATTGTGGGAAGAAGAAGAAGAAGGTCGGAGATGCTCAA1082Arg7TermGAAGAACGCGTTGTCACTCGATCGCTTTCTCCATTTTTTCCGGCTAGA-TGAGATTAAAGATAAAATTCGGATCTGTTGTTTTCGATCGAGTGACAACGC1083GCGTTGTCACTCGATCG10842,4-DB resistanceACAACAGATCCGAATTTTATCTTTAATCAGCCGGAAAAAATGGAGA10853-ketoacyl-CoAAAGCGATCGAGAGATAACGCGTTCTTCTTGAGCATCTCCGACCTTthiolaseCTTCTTCTTCTTCGCACAATTACGAGGCTTArabidopsis thalianaAAGCCTCGTAATTGTGCGAAGAAGAAGAAGAAGGTCGGAGATGC1086Gln8TermTCAAGAAGAACGCGTTATCTCTCGATCGCTTTCTCCATTTTTTCCGCAA-TAAGCTGATTAAAGATAAAATTCGGATCTGTTGTTCGAGAGATAACGCGTT1087AACGCGTTATCTCTCGA10882,4-DB resistanceGAGAGACAAAGAGTTCTTCTTGAACATCTCCGTCCTTCTTCTTCTT1089glyoxysomal beta-CCTCTCACAGCTTTTAAGGCTCTCTCTCTGCTTCAGCTTGCTTGGCketoacyol-thiolaseTGGGGACAGTGCTGCGTATCAGAGGACCTprecursorAGGTCGTCTGATACGCAGCACTGTCCCCAGCCAAGCAAGCTGAA1090Brassica napusGCAGAGAGAGAGCCTTAAAAGCTGTGAGAGGAAGAAGAAGAAGGGlu26TermACGGAGATGTTCAAGAAGAACTCTTTGTCTCTCGAA-TAAACAGCTTTTAAGGCTCT1091AGAGCCTTAAAAGCTGT10922,4-DB resistanceTTGAACATCTCCGTCCTTCTTCTTCTTCCTCTCACAGCTTTGAAGG1093glyoxysomal beta-CTCTCTCTCTGCTTGAGCTTGCTTGGCTGGGGACAGTGCTGCGTAketoacyol-thiolaseTCAGAGGACCTCTCTCTATGGAGATGATGTprecursorACATCATCTCCATAGAGAGAGGTCCTCTGATACGCAGCACTGTCC1094Brassica napusCCAGCCAAGCAAGCTCAAGCAGAGAGAGAGCCTTCAAAGCTGTGSer32TermAGAGGAAGAAGAAGAAGGACGGAGATGTTCAATCA-TGACTCTGCTTGAGCTTGCT1095AGCAAGCTCAAGCAGAG10962,4-DB resistanceTCTCCGTCCTTCTTCTTCTTCCTCTCACAGCTTTGAAGGCTCTCTC1097glyoxysomal beta-TCTGCTTCAGCTTGATTGGCTGGGGACAGTGCTGCGTATCAGAGketoacyol-thiolaseGACCTCTCTCTATGGAGATGATGTAGTCATTprecursorAATGACTACATCATCTCCATAGAGAGAGGTCCTCTGATACGCAGC1098Brassica napusACTGTCCCCAGCCAATCAAGCTGAAGCAGAGAGAGAGCCTTCAAACys34TermGCTGTGAGAGGAAGAAGAAGAAGGACGGAGATGC-TGATCAGCTTGATTGGCTGG1099CCAGCCAATCAAGCTGA11002,4-DB resistanceTCCGTCCTTCTTCTTGTTCCTCTCACAGCTTTGAAGGCTCTCTCTC1101glyoxysomal beta-TGCTTCAGCTTGCTAGGCTGGGGACAGTGCTGCGTATCAGAGGAketoacyol-thiolaseCCTCTCTCTATGGAGATGATGTAGTCATTGTprecursorACAATGACTACATCATCTCCATAGAGAGAGGTCGTCTGATACGCA1102Brassica napusGCACTGTCCCCAGCCTAGCAAGCTGAAGCAGAGAGAGAGCCTTCLeu35TermAAAGCTGTGAGAGGAAGAAGAAGAAGGACGGATTG-TAGAGCTTGCTAGGCTGGGG1103CCCCAGCCTAGCAAGCT11042,4-DB resistanceTCACAGCTTTGAAGGCTCTCTCTCTGCTTCAGCTTGCTTGGCTGG1105glyoxysomal beta-GGACAGTGCTGCGTAGCAGAGGACCTCTCTCTATGGAGATGATGTketoacyol-thiolaseAGTCATTGTTGCGGCACATAGGACTGCACTAprecursorTAGTGCAGTCCTATGTGCCGCAACAATGACTACATCATCTCCATA1106Brassica napusGAGAGAGGTCGTCTGCTACGCAGCACTGTCCCCAGCCAAGCAAGTyr42TermCTGAAGCAGAGAGAGAGCCTTCAAAGCTGTGATAT-TAGGCTGCGTAGCAGAGGAC1107GTCCTCTGCTACGCAGC11082,4-DB resistanceCAACAGACAGGAAGTGTTGCTCCAGCATCTCCGCCCTTCTAATTC11093-ketoacyl-CoATTCTTCTCACAATTAGee GAGTCCGCTCTTGCCGCATCAGTATGTGCTthiolase BGCAGGGGATAGCGCCGCATATCATAGGGCTMangifera indicaAGCCCTATGATATGCGGCGCTATCCCCTGCAGCACATACTGATGC1110Tyr25TermGGCAAGAGCGGACTCCTAATTGTGAGAAGAAGAATTAGAAGGGCTAC-TAGGGAGATGCTGGAGCAACACTTGCTGTCTGTTGCACAATTAGGAGTCCGC1111GCGGACTCCTAATTGTG11122,4-DB resistanceAACAGACAGCAAGTGTTGCTCCAGCATCTCCGCCCTTCTAATTCTT11133-ketoacyol-CoACTTCTCACAATTACTAGTCCGCTCTTGCCGCATCAGTATGTGCTGCthiolase BAGGGGATAGCGCCGCATATCATAGGGCTTMagnifera indicaAAGCCCTATGATATGCGGCGCTATCCCCTGCAGCACATACTGATG1114Glu26TermCGGCAAGAGCGGACTAGTAATTGTGAGAAGAAGAATTAGAAGGGGAG-TAGCGGAGATGCTGGAGCAACACTTGCTGTCTGTTACAATTACTAGTCCGCT1115AGCGGACTAGTAATTGT11162,4-DB resistanceTCCAGCATCTCCGCCCTTCTAATTCTTCTTCTCACAATTACGAGTC11173-ketoacy\to-CoACGCTCTTGCCGCATGAGTATGTGCTGCAGGGGATAGCGCCGCATthioblase BATCATAGGGCTTCTGTTTATGGAGACGATGTMangifera indicaACATCGTCTCCATAAACAGAAGCCCTATGATATGCGGCGCTATCC1118Ser32TermCCTGCAGCACATACTCATGCGGCAAGAGCGGACTCGTAATTGTGATCA-TGAGAAGAAGAATTAGAAGGGCGGAGATGCTGGATGCCGCATGAGTATGTG1119CACATACTCATGCGGCA11202,4-DB resistanceTCTCCGCCCTTCTAATTCTTCTTCTCACAATTACGAGTCCGCTCTT11213-ketoacyl-CoAGCCGCATCAGTATGAGCTGCAGGGGATAGCGCCGGATATCATAGthiolase BGGCTTCTGTTTATGGAGACGATGTGGTGATTMangifera indicaAATCACCACATCGTCTCCATAAACAGAAGCCCTATGATATGCGGC1122Cys34TermGCTATCCCCTGCAGCTCATACTGATGCGGCAAGAGCGGACTCGTTGT-TGAAATTGTGAGAAGAAGAATTAGAAGGGCGGAGATCAGTATGAGCTGCAGG1123CCTGCAGCTCATACTGA11242,4-DB resistanceTCACAATTACGAGTCCGCTCTTGCCGCATCAGTATGTGCTGCAGG11253-ketoacyl-CoAGGATAGCGCCGCATAGCATAGGGCTTGTGTTTATGGAGACGATGTthiolase BGGTGATTGTGGCAGGTCATCGTACTGCACTTMangifera indicaAAGTGCAGTAGGATGAGCTGCCACAATCACCACATCGTCTCCATA1126Tyr42TermAACAGAAGCCCTATGCTATGCGGCGCTATCCCCTGCAGCACATACTAT-TAGTGATGCGGCAAGAGCGGACTCGTAATTGTGAGCCGCATAGCATAGGGC1127GCCCTATGCTATGCGGC11282,4-DB resistanceGAAGGCGATCAACAGGCAGAGCATTTTGCTACATCATCTCCGGCC11293-ketoacyl-CoATTCTTCTTCCGCTTAGACAAATGAATCTTCGCTCTCTGCATCGGTTthiolaseTGTGCAGCTGGGGATAGTGCTTCGTATCAACucumis sativusTTGATACGAAGCACTATCCCCAGCTGCACAAACCGATGCAGAGAG1130Tyr22TermCGAAGATTCATTTGTCTAAGCGGAAGAAGAAGGCCGGAGATGATGTAG-TAGTAGCAAAATGCTCTGGCTGTTGATCGCCTTCTCCGCTTAGACAAATGA1131TCATTTGTCTAAGCGGA11322,4-DB resistanceATCAACAGGCAGAGCATTTTGCTACATCATCTCCGGCCTTCTTCTT11333-ketoacyl-CoACCGCTTACACAAATTAATCTTCGCTCTCTGCATCGGTTTGTGCAGCthiolaseTGGGGATAGTGCTTCGTATCAAAGGACATCucumis sativusATGTCCTTTGATACGAAGCAGTATCCCCAGCTGCACAAACCGATG1134Glu25TermCAGAGAGCGAAGATTAATTTGTGTAAGCGGAAGAAGAAGGCCGGGAA-TAAAGATGATGTAGCAAAATGCTCTGCCTGTTGATACACAAATTAATCTTCG1135CGAAGATTAATTTGTGT11362,4-DB resistanceGGCAGAGCATTTTGCTACATCATCTCCGGCCTTCTTCTTCCGCTTA11373-ketoacyl-CoACACAAATGAATCTTAGCTCTCTGCATCGGTTTGTGCAGCTGGGGAthiolaseTAGTGCTTCGTATCAAAGGACATCGGTGTTCucumis sativusAACACCGATGTCCTTTGATACGAAGCACTATCCCCAGCTGCACAA1138Ser27TermACCGATGCAGAGAGCTAAGATTCATTTGTGTAAGCGGAAGAAGAATCG-TAGGGCCGGAGATGATGTAGCAAAATGCTCTGCCTGAATCTTAGCTCTCTG1139CAGAGAGCTAAGATTCA11402,4-DB resistanceTGCTACATCATCTCCGGCCTTCTTCTTCCGCTTACACAAATGAATC11413-ketoacyl-CoATTCGCTCTCTGCATAGGTTTGTGCAGCTGGGGATAGTGCTTCGTAthiolaseTCAAAGGACATCGGTGTTTGGAGATGATGTCucumis sativusACATCATCTCCAAACACCGATGTCCTTTGATACGAAGCACTATCCC1142Ser31TermCAGCTGCACAAACCTATGCAGAGAGCGAAGATTCATTTGTGTAAGTCG-TAGCGGAAGAAGAAGGCCGGAGATGATGTAGCACTCTGCATAGGTTTGTG1143CACAAACCTATGCAGAG11442,4-DB resistanceTCATCTCCGGCCTTCTTCTTCCGCTTACACAAATGAATCTTCGCTC11453-ketoacyl-CoATCTGCATCGGTTTGAGCAGCTGGGGATAGTGCTTCGTATCAAAGGthiolaseACATCGGTGTTTGGAGATGATGTCGTGATTCucumis sativusAATCACGACATCATCTCCAAACACCGATGTCCTTTGATACGAAGCA1146Cys33TermCTATCCCCAGCTGCTCAAACCGATGCAGAGAGCGAAGATTCATTTTGT-TGAGTGTAAGCGGAAGAAGAAGGCCGGAGATGATCGGTTTGAGCAGCTGG1147CCAGCTGCTCAAACCGA11482A-DB resistanceGAAGGCAATCAACAGGCAGAGCATTCTGCTACATCATCTCCGGCC11493-ketoacyl-CoATTCATCTTCGGCTTAGACCCATGAATCTTCGCTCTCTGCATCGGTTthiolaseTGTGCAGCTGGGGATAGTGCGTCGTATCAACucurbita sp.TTGATACGACGCACTATCCCCAGCTGCACAAACCGATGCAGAGAG1150Tyr22TermCGAAGATTCATGGCTCTAAGCCGAAGATGAAGGCCGGAGATGATTAT-TAGGTAGCAGAATGCTCTGCCTGTTGATTGCCTTCTCGGCTTAGAGCCATGA1151TCATGGCTCTAAGCCGA11522,4-DB resistanceATCAACAGGCAGAGCATTCTGCTACATCATCTCCGGCCTTCATCTT11533-ketoacyl-CoACGGCTTATAGCCATTAATCTTCGCTCTCTGCATCGGTTTGTGCAGCthiolaseTGGGGATAGTGCGTCGTATCAAAGAACGTCucurbita sp.ACGTTCTTTGATACGACGCACTATCCCCAGCTGCACAAACCGATG1154Glu25TermCAGAGAGCGAAGATTAATGGCTATAAGCCGAAGATGAAGGCCGGGAA-TAAAGATGATGTAGCAGAATGCTCTGCCTGTTGATATAGCCATTAATCTTCG1155CGAAGATTAATGGCTAT11562,4-DB resistanceGGCAGAGCATTCTGCTACATCATCTCCGGCCTTCATCTTCGGCTT11573-ketoacyl-CoAATAGCCATGAATCTTAGCTCTCTGCATCGGTTTGTGCAGCTGGGGthiolaseATAGTGCGTCGTATCAAAGAACGTCGGTGTTCucurbita sp.AACACCGACGTTCTTTGATACGACGCACTATCCCCAGCTGCACAA1158Ser27TermACCGATGCAGAGAGCTAAGATTCATGGCTATAAGCCGAAGATGAATCG-TAGGGCCGGAGATGATGTAGCAGAATGCTCTGCCTGAATCTTAGCTCTCTG1159CAGAGAGCTAAGATTCA11602,4-DB resistanceTGCTACATCATCTCCGGCCTTCATCTTCGGCTTATAGCCATGAATC11613-ketoacyl-CoATTCGCTCTCTGCATAGGTTTGTGCAGCTGGGGATAGTGCGTCGTAthiolaseTCAAAGAACGTCGGTGTTTGGAGATGATGTCucurbita sp.ACATCATCTCCAAACACCGACGTTCTTTGATACGACGCACTATCCC1162Ser31TermCAGCTGCACAAACCTATGCAGAGAGCGAAGATTCATGGCTATAAGTCG-TAGCCGAAGATGAAGGCCGGAGATGATGTAGCACTCTGCATAGGTTTGTG1163CACAAACCTATGCAGAG11642,4-DB resistanceTCATCTCCGGCCTTCATCTTCGGCTTATAGCCATGAATCTTCGCTC11653-ketoacyl-CoATCTGCATCGGTTTGAGCAGCTGGGGATAGTGCGTCGTATCAAAGAthiolaseACGTCGGTGTTTGGAGATGATGTCGTGATACucurbita sp.TATCACGACATCATCTCCAAACACCGACGTTCTTTGATACGACGCA1166Cys33TermCTATCCCCAGCTGCTCAAACCGATGCAGAGAGCGAAGATTCATGGTGT-TGACTATAAGCCGAAGATGAAGGCCGGAGATGATCGGTTTGAGCAGCTGG1167CCAGCTGCTCAAACCGA11682,4 DB resistanceTCATAGTCTCTTTTGCCGCTTGGATTCTTCCAAGGTTAGTGAGCTG1169Pex14CTATGGCAACTCATTAGCAAACGCAACCTCCTTCCGATTTTCCCGCArabidopsis thalianaTCTTGCCGATGAAAATTCCCAGATTCCAGGln5TermCTGGAATCTGGGAATTTTCATCGGCAAGAGCGGGAAAATCGGAA1170CAG-TAGGGAGGTTGCGTTTGCTAATGAGTTGCCATAGCAGCTCACTAACCTTGGAAGAATCCAAGCGGCAAAAGAGACTATGACAACTCATTAGCAAACG1171CGTTTGCTAATGAGTTG11722,4 DB resistanceTAGTCTCTTTTGCCGCTTGGATTCTTCCAAGGTTAGTGAGCTGCTA1173Pex14TGGCAACTCATCAGTAAACGCAACCTCCTTCCGATTTTCCCGCTCTArabidopsis thalianaTGCCGATGAAAATTCCCAGATTGCAGGTTGln6TermAACCTGGAATCTGGGAATTTTCATCGGCAAGAGCGGGAAAATCGG1174CAA-TAAAAGGAGGTTGCGTTTACTGATGAGTTGCCATAGCAGCTCACTAACCTTGGAAGAATCCAAGCGGCAAAAGAGACTACTCATCAGTAAACGCAA1175TTGCGTTTACTGATGAG11762,4 DB resistanceCTTTTGCCGCTTGGATTCTTCCAAGGTTAGTGAGCTGCTATGGCA1177Pex14ACTCATCAGCAAACGTAACCTCCTTCCGATTTTCCCGCTCTTGCCGArabidopsis thalianaATGAAAATTCCGAGATTCCAGGTTCAATTTGln8TermAAATTGAACCTGGAATCTGGGAATTTTCATCGGCAAGAGCGGGAA1178CAA-TAAAATCGGAAGGAGGTTACGTTTGCTGATGAGTTGCCATAGCAGCTCACTAACCTTGGAAGAATCCAAGCGGCAAAAGAGCAAACGTAACCTCCT1179AGGAGGTTACGTTTGCT11802,4 DB resistanceGCTGCTATGGCAACTGATGAGCAAACGCAACCTCCTTCCGATTTT1181Pex14CCCGCTCTTGCCGATTAAAATTCCCAGATTCCAGGTTCAATTTACAArabidopsis thalianaCCTTCTAATCATTATTTCTTAATTTTTCTTGlu19TermAAGAAAAATTAAGAAATAATGATTAGAAGGTGTAAATTGAACCTGG1182GAA-TAAAATCTGGGAATTTTAATCGGCAAGAGCGGGAAAATCGGAAGGAGGTTGCGTTTGCTGATGAGTTGCCATAGCAGCTTGCCGATTAAAATTCC1183GGAATTTTAATCGGCAA11842,4 DB resistanceGCAACTCATCAGCAAACGCAACCTCCTTCCGATTTTCCCGCTCTT1185Pex14GCCGATGAAAATTCCTAGATTCCAGGTTCAATTTACACCTTCTAATArabidopsis thalianaCATTATTTCTTAATTTTTCTTTGGTGGATTGln22TermAATCCACCAAAGAAAAATTAAGAAATAATGATTAGAAGGTGTAAAT1186CAG-TAGTGAACCTGGAATCTAGGAATTTTCATCGGCAAGAGCGGGAAAATCGGAAGGAGGTTGCGTTTGCTGATGAGTTGCAAAATTCCTAGATTCCA1187TGGAATCTAGGAATTTT1188

EXAMPLE 8

Production of Albino Mutants for the Analysis of Photosynthetic Processes

[0132] Plant productivity is limited by resources available and the ability of plants to harness these resources. The conversion of light to chemical energy, which is then used to synthesize carbohydrates, fatty acids, sugars, amino acids and other compounds, requires a complex system which combines the light harvesting apparatus of pigments and proteins. The value of light energy to the plant can only be realized when it is efficiently converted into chemical energy by photosynthesis and fed into various biochemical processes. Significant effort has therefore been directed at studying photosynthetic processes in plants in order to improve productivity and/or the efficiency of photosynthesis. The analysis of the photosynthetic process is substantially aided by the ability to produce albino plants.

[0133] The attached table discloses exemplary oligonucleotide base sequences which can be used to generate site-specific mutations in genes involved in starch metabolism.

20TABLE 18Oligonucleotides to produce albino plantsPhenotype, Gene,Plant & TargetedSEQ IDAlterationAltering OligosNO:White leavesTTCTTTCCTGTGAAATTATCTGCTCAAATCTTTGGTTCCTGACGGAG1189ImmutansATGGCGGCGATTTGAGGCATCTCCTCTGGTACGTTGACGATTTCAArabidopsis thalianaCGGCCTTTGGTTACTCTTCGACGCTCTAGSer5TermCTAGAGCGTCGAAGAGTAACCAAAGGCCGTGAAATCGTCAACGTA1190TCA-TGACCAGAGGAGATGCCTCAAATCGCCGCCATCTCCGTCAGGAACCAAAGATTTGAGCAGATAATTTCACAGGAAAGAAGGCGATTTGAGGCATCT1191AGATGCCTCAAATCGCC1192White leavesGCTCAAATCTTTGGTTCCTGACGGAGATGGCGGCGATTTCAGGCA1193ImmutansTCTCCTCTGGTACGTAGACGATTTCACGGCCTTTGGTTACTCTTCGArabidopsis thalianaACGCTCTAGAGCCGCCGTTTCGTACAGCTCLeu12TermGAGCTGTACGAAACGGCGGCTCTAGAGCGTCGAAGAGTAACCAAA 1194TTG-TAGGGCCGTGAAATCGTCTACGTACCAGAGGAGATGCCTGAAATCGCCGCCATCTCCGTCAGGAACCAAAGATTTGAGCTGGTACGTAGACGATTT1195AAATCGTCTACGTACCA1196White leavesTTTGGTTCCTGACGGAGATGGCGGCGATTTCAGGCATCTCCTCTG1197ImmutansGTACGTTGACGATTTGACGGCCTTTGGTTACTCTTCGACGCTCTAGArabidopsis thalianaAGCCGCCGTTTCGTACAGCTCCTCTCACCGSer15TermCGGTGAGAGGAGCTGTACGAAACGGCGGCTCTAGAGCGTCGAAG1198TCA-TGAAGTAACCAAAGGCCGTCAAATCGTCAACGTACCAGAGGAGATGCCTGAAATCGCCGCCATCTCCGTCAGGAACCAAAGACGATTTGACGGCCTT1199AAGGCCGTCAAATCGTC1200White leavesGCGGCGATTTCAGGCATCTCCTCTGGTACGTTGACGATTTCACGG1201ImmutansCCTTTGGTTACTCTTTGACGCTCTAGAGCCGCCGTTTCGTACAGCTArabidopsis thalianaCCTCTCACCGATTGCTTCATCATCTTCCTCArg22TermGAGGAAGATGATGAAGCAATCGGTGAGAGGAGCTGTACGAAACG1202CGA-TGAGCGGCTCTAGAGCGTCAAAGAGTAACCAAAGGCCGTGAAATCGTCAACGTACCAGAGGAGATGCCTGAAATCGCCGCTTACTCTTTGACGCTCT1203AGAGCGTCAAAGAGTAA1204White leavesTCAGGCATCTCCTCTGGTACGTTGACGATTTCACGGCCTTTGGTTA1205ImmutansCTCTTCGACGCTCTTGAGCCGCCGTTTCGTACAGCTCCTCTCACCArabidopsis thalianaGATTGCTTCATCATCTTCCTCTCTCTTCTCArg25TermGAGAAGAGAGAGGAAGATGATGAAGCAATCGGTGAGAGGAGCTG1206AGA-TGATACGAAACGGCGGCTCAAGAGCGTCGAAGAGTAACCAAAGGCCGTGAAATCGTCAACGTACCAGAGGAGATGCCTGAGACGCTCTTGAGCCGCC1207GGCGGCTCAAGAGCGTC1208White leavesGATTCTTGTGGGAAGGAAGAAGGATCAAGAATGGCGATTTCGATT1209ImmutansTCTGCTATGAGTTTTTGAACCTCAGTTTCTTCATATTCTTGTTTTAGLycopersiconAGCTAGGAGTTTTGAGAAGTCATCAGTTTesculentumAAACTGATGACTTCTCAAAACTCCTAGCTCTAAAACAAGAATATGA1210Gly11TermAGAAACTGAGGTTCAAAAACTCATAGCAGAAATCGAAATCGCCATTGGA-TGACTTGATCCTTCTTCCTTCCCACAAGAATCTGAGTTTTTGAACCTCA1211TGAGGTTCAAAAACTCA1212White leavesGTGGGAAGGAAGAAGGATCAAGAATGGCGATTTCGATTTCTGCTA1213ImmutansTGAGTTTTGGAACCTGAGTTTCTTCATATTCTTGTTTTAGAGCTAGGLycopersiconAGTTTTGAGAAGTCATCAGTTTTATGCAAesculentumTTGCATAAAACTGATGACTTCTCAAAACTCCTAGCTCTAAAACAAG1214Ser13TermAATATGAAGAAACTCAGGTTCCAAAACTCATAGCAGAAATCGAAATTCA-TGACGCCATTCTTGATCCTTCTTCCTTCCCACTGGAACCTGAGTTTCTT1215AAGAAACTCAGGTTCCA1216White leavesAAGAAGGATCAAGAATGGCGATTTCGATTTCTGCTATGAGTTTTGG1217ImmutansAACCTCAGTTTCTTGATATTCTTGTTTTAGAGCTAGGAGTTTTGAGALycopersiconAGTCATCAGTTTTATGCAATTCCCAGAAesculentumTTCTGGGAATTGCATAAAACTGATGACTTCTCAAAACTCCTAGCTC1218Ser16TermTAAAACAAGAATATCAAGAAACTGAGGTTCCAAAACTCATAGCAGATCA-TGAAATCGAAATCGCCATTCTTGATCCTTCTTAGTTTCTTGATATTCTT1219AAGAATATCAAGAAACT1220White leavesAGGATCAAGAATGGCGATTTCGATTTCTGCTATGAGTTTTGGAACC1221ImmutansTCAGTTTCTTCATAGTCTTGTTTTAGAGCTAGGAGTTTTGAGAAGTCLycopersiconATCAGTTTTATGCAATTCCCAGAACCCAesculentumTGGGTTCTGGGAATTGCATAAAACTGATGACTTCTCAAAACTCCTA1222Tyr17TermGCTCTAAAACAAGACTATGAAGAAACTGAGGTTCCAAAACTCATAGTAT-TAGCAGAAATCGAAATCGCCATTCTTGATCCTTCTTCATAGTCTTGTTT1223AAACAAGACTATGAAGA1224White leavesAAGAATGGCGATTTCGATTTCTGCTATGAGTTTTGGAACCTCAGTT1225ImmutansTCTTCATATTCTTGATTTAGAGCTAGGAGTTTTGAGAAGTCATCAGTLycopersiconTTTATGCAATTCCCAGAACCCATGTCGGesculentumCCGACATGGGTTCTGGGAATTGCATAAAACTGATGACTTCTCAAAA1226Cys19TermCTCCTAGCTCTAAATCAAGAATATGAAGAAACTGAGGTTCCAAAACTGT-TGATCATAGCAGAAATCGAAATCGCCATTCTTTATTCTTGATTTAGAGC1227GCTCTAAATCAAGAATA1228White leavesCGCGTCCGATAAAAAAATCAAGAATGGCGATTTCCATATCTGCTAT1229ImmutansGAGTTTTCGAACTTGAGTTTCTTCTTCATATTCAGCATTTTTGTGCACapsicum annuumATTCCAAGAACCCATTTTGTTTGAATTCSer13TermGAATTCAAACAAAATGGGTTCTTGGAATTGCACAAAAATGCTGAAT1230TCA-TGAATGAAGAAGAAACTCAAGTTCGAAAACTCATAGCAGATATGGAAATCGCCATTCTTGATTTTTTTATCGGACGCGTCGAACTTGAGTTTCTT1231AAGAAACTCAAGTTCGA1232White leavesAAAAATCAAGAATGGCGATTTCCATATCTGCTATGAGTTTTCGAAC1233ImmutansTTCAGTTTCTTCTTGATATTCAGCATTTTTGTGCAATTCCAAGAACCCapsicum annuumCATTTTGTTTGAATTCTCTATTTTCACTSer17TermAGTGAAAATAGAGAATTCAAACAAAATGGGTTCTTGGAATTGCACA1234TCA-TGAAAAATGCTGAATATCAAGAAGAAACTGAAGTTCGAAAACTCATAGCAGATATGGAAATCGCCATTCTTGATTTTTTTCTTCTTGATATTCAG1235CTGAATATCAAGAAGAA1236White leavesCAAGAATGGCGATTTCCATATCTGCTATGAGTTTTCGAACTTCAGT1237ImmutansTTCTTCTTCATATTGAGCATTTTTGTGCAATTCCAAGAACCCATTTTCapsicum annuumGTTTGAATTCTCTATTTTCACTTAGGAASer19TermTTCCTAAGTGAAAATAGAGAATTCAAACAAAATGGGTTCTTGGAAT1238TCA-TGATGCACAAAAATGCTCAATATGAAGAAGAAACTGAAGTTCGAAAACTCATAGCAGATATGGAAATCGCCATTCTTGTTCATATTGAGCATTTT1239AAAATGCTCAATATGAA1240White leavesCGATTTCCATATCTGCTATGAGTTTTCGAACTTCAGTTTCTTCTTCA1241ImmutansTATTCAGCATTTTAGTGCAATTCCAAGAACCCATTTTGTTTGAATTCCapsicum annuumTCTATTTTCACTTAGGAATTCTCATAGLeu21TermCTATGAGAATTCCTAAGTGAAAATAGAGAATTCAAACAAAATGGGT1242TTG-TAGTCTTGGAATTGCACTAAAATGCTGAATATGAAGAAGAAACTGAAGTTCGAAAACTCATAGCAGATATGGAAATCGAGCATTTTAGTGCAATT1243AATTGCACTAAAATGCT1244White leavesTTCCATATCTGCTATGAGTTTTCGAACTTCAGTTTCTTCTTCATATT1245ImmutansCAGCATTTTTGTGAAATTCCAAGAACCCATTTTGTTTGAATTCTCTACapsicum annuumTTTTCACTTAGGAATTCTCATAGAACTCys22TermAGTTCTATGAGAATTCCTAAGTGAAAATAGAGAATTCAAACAAAAT1246TGC-TGAGGGTTCTTGGAATTTCACAAAAATGCTGAATATGAAGAAGAAACTGAAGTTCGAAAACTCATAGCAGATATGGAATTTTTGTGAAATTCCAA1247TTGGAATTTCACAAAAA1248White leavesTTCGGCACGAGGGAGAAGGAGCAGACCGAGGTGGCCGTCGAGG1249ImmutansAGTCCTTCCCCTTCAGGTAGACGGCTCCTCCTGACGAGCCACTGGOryza sativaTCACCGCCGAGGAGAGCTGGGTGGTTAAGCTCGGlu22TermCGAGCTTAACCACCCAGCTCTCCTCGGCGGTGACCAGTGGCTCGT1250GAG-TAGCAGGAGGAGCCGTCTACCTGAAGGGGAAGGACTCCTCGACGGCCACCTCGGTCTGCTCCTTCTCCCTCGTGCCGAACCTTCAGGTAGACGGCT1251AGCCGTCTACCTGAAGG1252White leavesGAGCAGACCGAGGTGGCCGTCGAGGAGTCCTTCCCCTTCAGGGA1253ImmutansGACGGCTCCTCCTGACTAGCCACTGGTCACCGCCGAGGAGAGCTOryza sativaGGGTGGTTAAGCTCGAGCAGTCCGTGAACATTTGlu28TermAAATGTTCACGGACTGCTCGAGCTTAACCACCCAGCTCTCCTCGG1254CAG-TAGCGGTGACCAGTGGCTAGTCAGGAGGAGCCGTCTCCCTGAAGGGGAAGGACTCCTCGACGGCCACCTCGGTCTGCTCCTCCTGACTAGCCACTG1255CAGTGGCTAGTCAGGAG1256White leavesGTCGAGGAGTCCTTCCCCTTCAGGGAGACGGCTCCTCCTGACGA1257ImmutansGCCACTGGTCACCGCCTAGGAGAGCTGGGTGGTTAAGCTCGAGCOryza sativaAGTCCGTGAACATTTTCCTCACGGAGTCAGTCAGlu34TermTGACTGACTCCGTGAGGAAAATGTTCACGGACTGCTCGAGCTTAA1258GAG-TAGCCACCCAGCTCTCCTAGGCGGTGACCAGTGGCTCGTCAGGAGGAGCCGTCTCCCTGAAGGGGAAGGACTCCTCGACTCACCGCCTAGGAGAGC1259GCTCTCCTAGGCGGTGA1260White leavesGAGGAGTCCTTCCCCTTCAGGGAGACGGCTCCTCCTGACGAGCC1261ImmutansACTGGTCACCGCCGAGTAGAGCTGGGTGGTTAAGCTCGAGCAGTOryza sativaCCGTGAACATTTTCCTCACGGAGTCAGTCATCAGlu35TermTGATGACTGACTCCGTGAGGAAAATGTTCACGGACTGCTCGAGCT1262GAG-TAGTAACCACCCAGCTCTACTCGGCGGTGACCAGTGGCTCGTCAGGAGGAGCCGTCTCCCTGAAGGGGAAGGACTCCTCCCGCCGAGTAGAGCTGG1263CCAGCTCTACTCGGCGG1264White leavesCTTCCCCTTCAGGGAGACGGCTCCTCCTGACGAGCCACTGGTCAC1265ImmutansCGCCGAGGAGAGCTGAGTGGTTAAGCTCGAGCAGTCCGTGAACAOryza sativaTTTTCCTCACGGAGTCAGTCATCACGATACTTTrp37TermAAGTATCGTGATGACTGACTCCGTGAGGAAAATGTTCACGGACTG1266TGG-TGACTCGAGCTTAACCACTCAGCTCTCCTCGGCGGTGACCAGTGGCTCGTCAGGAGGAGCCGTCTCCCTGAAGGGGAAGGAGAGCTGAGTGGTTAA1267TTAACCACTCAGCTCTC1268White leavesTCCGGAGGAGGAAGGGGGATTCGACGAGGAGCTCACCCTCGCCG1269ImmutansGCGAGGACGGCGACTGAGTCGTCAGATTCGAGCAGTCCTTCAACTriticum aestivumGTATTCCTCACGGATACTGTCATCTTTATACTCTrp22TermGAGTATAAAGATGACAGTATCCGTGAGGAATACGTTGAAGGACTG1270TGG-TGACTCGAATCTGACGACTCAGTCGCCGTCCTCGCCGGCGAGGGTGAGCTCCTCGTCGAATCCCCCTTCCTCCTCCGGAGGCGACTGAGTCGTCAG1271CTGACGACTCAGTCGCC1272White leavesGAGGAAGGGGGATTCGACGAGGAGCTCACCCTCGCCGGCGAGG1273ImmutansACGGCGACTGGGTCGTCTGATTCGAGCAGTCCTTCAACGTATTCCTriticum aestivumTCACGGATACTGTCATCTTTATACTCGATATTCArg25TermGAATATCGAGTATAAAGATGACAGTATCCGTGAGGAATACGTTGAA1274AGA-TGAGGACTGCTCGAATCAGACGACCCAGTCGCCGTCCTCGCCGGCGAGGGTGAGCTCCTCGTCGAATCCCCCTTCCTCGGGTCGTCTGATTCGAG1275CTCGAATCAGACGACCC1276White leavesGGGGGATTCGACGAGGAGCTCACCCTCGCCGGCGAGGACGGCG1277ImmutansACTGGGTCGTCAGATTCTAGCAGTCCTTCAACGTATTCCTCACGGATriticum aestivumTACTGTCATCTTTATACTCGATATTCTGTATCGlu21TermGATACAGAATATCGAGTATAAAGATGACAGTATCCGTGAGGAATAC1278GAG-TAGGTTGAAGGACTGCTAGAATCTGACGACCCAGTCGCCGTCCTCGCCGGCGAGGGTGAGCTCCTCGTCGAATCCCCCTCAGATTCTAGCAGTCC1279GGACTGCTAGAATCTGA1280White leavesGGATTCGACGAGGAGCTCACCCTCGCCGGCGAGGACGGCGACTG1281ImmutansGGTCGTCAGATTCGAGTAGTCCTTCAACGTATTCCTCACGGATACTTriticum aestivumGTCATCTTTATACTCGATATTCTGTATCGTGGln28TermCACGATACAGAATATCGAGTATAAAGATGACAGTATCCGTGAGGAA1282CAG-TAGTACGTTGAAGGACTACTCGAATCTGACGACCCAGTCGCCGTCCTCGCCGGCGAGGGTGAGCTCCTCGTCGAATCCGATTCGAGTAGTCCTTC1283GAAGGACTACTCGAATC1284White leavesCGAGCAGTCCTTCAACGTATTCCTCACGGATACTGTCATCTTTATA1285ImmutansCTCGATATTCTGTAGCGTGACCGCGACTACGCAAGGTTCTTCGTGTriticum aestivumCTCGAGACCATCGCCAGGGTGCCCTATTTCTyr46TermGAAATAGGGCACCCTGGCGATGGTCTCGAGCACGAAGAACCTTG1286TAT-TAGCGTAGTCGCGGTCACGCTACAGAATATCGAGTATAAAGATGACAGTATCCGTGAGGAATACGTTGAAGGACTGCTCGATTCTGTAGCGTGACCG1287CGGTCACGCTACAGAAT1288

EXAMPLE 9

Altering Amino Acid Content of Plants

[0134] Another aim of biotechnology is to generate plants, especially crop plants, with added value traits. An example of such a trait is improved nutritional quality in food crops. For example, lysine, tryptophan and threonine, which are essential amino acids in the diet of humans and many animals, are limiting nutrients in most cereal crops. Consequently, grain-based diets, such as those based on corn, barley, wheat, rice, maize, millet, sorghum, and the like, must be supplemented with more expensive synthetic amino acids or amino-acid-containing oilseed protein meals. Increasing the lysine content of these grains or of any of the feed component crops would result in significant added value.

[0135] Naturally occurring mutants of plants that have different levels of particular essential amino acids have been identified. However, these mutants are generally not the result of increased free amino acid, but are instead the result of shifts in the overall protein profile of the grain. For example, in maize, reduced levels of lysine-deficient endosperm proteins (prolamines) are complemented by elevated levels of more lysine-rich proteins (albumins, globulins and glutelins). While nutritionally superior, these mutants are associated with reduced yields and poor grain quality, limiting their agronomic usefulness.

[0136] An alternative approach is to generate plants with mutations that render key amino acid biosynthetic enzymes insensitive to feedback inhibition. Many such mutations are known and mutation results in increased free amino acid. The increased production can optionally be coupled to increased expression of an abundant storage protein comprising the chosen amino acid. Alternatively, a normally abundant protein can be engineered to contain more of the target amino acid.

[0137] The attached table discloses exemplary oligonucleotide base sequences which can be used to generate site-specific mutations that remove feedback inhibition in plant amino acid biosynthetic enzymes.

21TABLE 19Genome-Altering Oligos Conferring Amino Acid OverproductionPhenotype, Gene,Plant & TargetedSEQ IDAlterationAltering OligosNO:Met OverproductionTATCCTCCAGGATCTTAAGATTTCCTCCTAATTTCGTCCGTCAGCT1289CGSGAGCATTAAAGCCCATAGAAACTGTAGCAACATCGGTGTTGCACAArabidopsis thalianaGATCGTGGCGGCTAAGTGGTCCAACAACCCArg77HisGGGTTGTTGGACCACTTAGCCGCCACGATCTGTGCAACACCGAT1290CGT-CATGTTGCTACAGTTTCTATGGGCTTTAATGCTCAGCTGACGGACGAAATTAGGAGGAAATCTTAAGATCCTGGAGGATATAAAGCCCATAGAAACT1291AGTTTCTATGGGCTTTA1292Met OverproductionTCTTAAGATTTCCTCCTAATTTCGTCCGTCAGCTGAGCATTAAAGC1293CGSCCGTAGAAACTGTAACAACATCGGTGTTGCACAGATCGTGGCGGArabidopsis thalianaCTAAGTGGTCCAACAACCCATCCTCCGCGTTSer81AsnAACGCGGAGGATGGGTTGTTGGACCACTTAGCCGCCACGATCTG1294AGC-AACTGCAACACCGATGTTGTTACAGTTTCTACGGGCTTTAATGCTCAGCTGACGGACGAAATTAGGAGGAAATCTTAAGAAAACTGTAACAACATCG1295CGATGTTGTTACAGTTT1296Met OverproductionTTTCCTCCTAATTTCGTCCGTCAGCTGAGCATTAAAGCCCGTAGAA1297CGSACTGTAGCAACATCAGTGTTGCACAGATCGTGGCGGCTAAGTGGTArabidopsis thalianaCCAACAACCCATCCTCCGCGTTACCTTCGGGly84SerCCGAAGGTAACGCGGAGGATGGGTTGTTGGACCACTTAGCCGCC1298GGT-AGTACGATCTGTGCAACACTGATGTTGCTACAGTTTCTACGGGCTTTAATGCTCAGCTGACGGACGAAATTAGGAGGAAAGCAACATCAGTGTTGCA1299TGCAACACTGATGTTGC1300Met OverproductionTTCCTCCTAATTTCGTCCGTCAGCTGAGCATTAAAGCCCGTAGAAA1301CGSCTGTAGCAACATCGATGTTGCACAGATCGTGGCGGCTAAGTGGTCArabidopsis thalianaCAACAACCCATCCTCCGCGTTACCTTCGGCGly84AspGCCGAAGGTAACGCGGAGGATGGGTTGTTGGACCACTTAGCCGC1302GGT-GATCACGATCTGTGCAACATCGATGTTGCTACAGTTTCTACGGGCTTTAATGCTCAGCTGACGGACGAAATTAGGAGGAACAACATCGATGTTGCAC1303GTGCAACATCGATGTTG1304Met OverproductionTATCGTCACTCATCCTCCGCTTCCCTCCCAACTTCGTCCGCCAGC1305CGSTCAGCACCAAGGCCCACCGCAACTGCAGCAACATCGGCGTCGCGFragraria vescaCAGATCGTCGCGGCTTCGTGGTCCAACAAAGAArg73HisTCTTTGTTGGACCACGAAGCCGCGACGATCTGCGCGACGCCGAT1306CGC-CACGTTGCTGCAGTTGCGGTGGGCCTTGGTGCTGAGCTGGCGGACGAAGTTGGGAGGGAAGCGGAGGATGAGTGACGATACAAGGCCCACCGCAACT1307AGTTGCGGTGGGCCTTG1308Met OverproductionTCCTCCGCTTCCCTCCCAACTTCGTCCGCCAGCTCAGCACCAAGG1309CGSCCCGCCGCAACTGCAACAACATCGGCGTCGCGCAGATCGTCGCGFragraria vescaGCTTCGTGGTCCAACAAAGACTCCGACCTTTCSer77AsnGAAAGGTCGGAGTCTTTGTTGGACCACGAAGCCGCGACGATCTG1310AGC-AACCGCGACGCCGATGTTGTTGCAGTTGCGGCGGGCCTTGGTGCTGAGCTGGCGGACGAAGTTGGGAGGGAAGCGGAGGACAACTGCAACAACATCG1311CGATGTTGTTGCAGTTG1312Met OverproductionTTCCCTCCCAACTTCGTCCGCCAGCTCAGCACCAAGGCCCGCCG1313CGSCAACTGCAGCAACATCAGCGTCGCGCAGATCGTCGCGGCTTCGTFragraria vescaGGTCCAACAAAGACTCCGACCTTTCGGCGGTGCGly80SerGCACCGCCGAAAGGTCGGAGTCTTTGTTGGACCACGAAGCCGCG1314GGC-AGCACGATCTGCGCGACGCTGATGTTGGTGCAGTTGCGGCGGGCCTTGGTGCTGAGCTGGCGGACGAAGTTGGGAGGGAAGCAACATCAGCGTCGCG1315CGCGACGCTGATGTTGC1316Met OverproductionTCCCTCCCAACTTCGTCCGCCAGCTCAGCACCAAGGCCCGCCGC1317CGSAACTGCAGCAACATCGACGTCGCGCAGATCGTCGCGGCTTCGTGFragraria vescaGTCCAACAAAGACTCCGACCTTTCGGCGGTGCCGly80AspGGCACCGCCGAAAGGTCGGAGTCTTTGTTGGACCACGAAGCCGC1318GGC-GACGACGATCTGCGCGACGTCGATGTTGCTGCAGTTGCGGCGGGCCTTGGTGCTGAGCTGGCGGACGAAGTTGGGAGGGACAACATCGACGTCGCGC1319GCGCGACGTCGATGTTG1320Met OverproductionTCTCCTCCCTCATCCTCCGCTTCCCTCCCAACTTCCAGCGCCAGC1321CGSTAAGCACCAAGGCGAGCCGCAACTGCAGCAACATCGGCGTCGCGGlycine maxCAAATCGTCGCCGCTTCGTGGTCGAACAACAGArg68HisCTGTTGTTCGACCACGAAGCGGCGACGATTTGCGCGACGCCGAT1322CGC-CACGTTGCTGCAGTTGCGGCTCGCCTTGGTGCTTAGCTGGCGCTGGAAGTTGGGAGGGAAGCGGAGGATGAGGGAGGAGACCAAGGCGAGCCGCAAC1323GTTGCGGCTCGCCTTGG1324Met OverproductionTCCTCCGCTTCCCTCCCAACTTCCAGCGCCAGCTAAGCACCAAGG1325CGSCGCGCCGCAACTGCAACAACATCGGCGTCGCGCAAATCGTCGCCGlycine maxGCTTCGTGGTCGAACAACAGCGACAACTCTCCSer72AsnGGAGAGTTGTCGCTGTTGTTCGACCACGAAGCGGCGACGATTTG1326AGC-AACCGCGACGCCGATGTTGTTGCAGTTGCGGCGCGCCTTGGTGCTTAGCTGGCGCTGGAAGTTGGGAGGGAAGCGGAGGACAACTGCAACAACATCG1327CGATGTTGTTGCAGTTG1328Met OverproductionTTCCCTCCCAACTTCCAGCGCCAGCTAAGCACCAAGGCGCGCCG1329CGSCAACTGCAGCAACATCAGCGTCGCGCAAATCGTCGCCGCTTCGTGlycine maxGGTCGAACAACAGCGACAACTCTCCGGCCGCCGGly75SerCGGCGGCCGGAGAGTTGTCGCTGTTGTTCGACCACGAAGCGGCG1330GGC-AGCACGATTTGCGCGACGCTGATGTTGCTGCAGTTGCGGCGCGCCTTGGTGCTTAGCTGGCGCTGGAAGTTGGGAGGGAAGCAACATCAGCGTCGCG1331CGCGACGCTGATGTTGC1332Met OverproductionTCCCTCCCAACTTCCAGCGCCAGCTAAGCACCAAGGCGCGCCGC1333CGSAACTGCAGCAACATCGACGTCGCGCAAATCGTCGCCGCTTCGTGGlycine maxGTCGAACAACAGCGACAACTCTCCGGCCGCCGGGly75AspCCGGCGGCGGGAGAGTTGTCGCTGTTGTTCGACCACGAAGCGGC1334GGC-GACGACGATTTGCGCGACGTCGATGTTGCTGCAGTTGCGGCGCGCCTTGGTGCTTAGCTGGCGCTGGAAGTTGGGAGGGACAACATCGACGTCGCGC1335GCGCGACGTCGATGTTG1336Met OverproductionTGTCTTCTCTGATTTTCAGGTTTCCTCCTAATTTCGTGAGGCAGCT1337CGSAAGCATTAAGGCTCACAGGAATTGCAGCAATATTGGCGTGGCTCASolanum tuberosumAGTTGTGGCGGCTTCCTGGTCTAACAACCAArg70HisTGGTTGTTAGACCAGGAAGCCGCCACAACTTGAGCCACGCCAATA1338AGG-CACTTGCTGCAATTCCTGTGAGCCTTAATGCTTAGCTGCCTCACGAAATTAGGAGGAAACCTGAAAATCAGAGAAGACATAAGGCTCACAGGAATT1339AATTCCTGTGAGCCTTA1340Met OverproductionTTTTCAGGTTTCCTCCTAATTTCGTGAGGCAGCTAAGCATTAAGGC1341CGSTAGGAGGAATTGCAACAATATTGGCGTGGCTCAAGTTGTGGCGGSolanum tuberosumCTTCCTGGTCTAACAACCAAGCCGGTCCTGASer74AsnTCAGGACCGGCTTGGTTGTTAGACCAGGAAGCCGCCACAACTTG1342AGC-AACAGCCACGCCAATATTGTTGCAATTCCTCCTAGCCTTAATGCTTAGCTGCCTCACGAAATTAGGAGGAAACCTGAAAAGAATTGCAACAATATTG1343CAATATTGTTGCAATTC1344Met OverproductionTTTCCTCCTAATTTCGTGAGGCAGCTAAGCATTAAGGCTAGGAGG1345CGSAATTGCAGCAATATTAGCGTGGCTCAAGTTGTGGCGGCTTCCTGGSolanum tuberosumTCTAACAACCAAGCCGGTCCTGAATTCACTCGly77SerGAGTGAATTCAGGACCGGCTTGGTTGTTAGACCAGGAAGCCGCC1346GGC-AGCACAACTTGAGCCACGCTAATATTGCTGCAATTCCTCCTAGCCTTAATGCTTAGCTGCCTCACGAAATTAGGAGGAAAGCAATATTAGCGTGGGT1347AGCCACGCTAATATTGC1348Met OverproductionTTCCTCCTAATTTCGTGAGGCAGCTAAGCATTAAGGCTAGGAGGA1349CGSATTGCAGCAATATTGACGTGGCTCAAGTTGTGGCGGCTTCCTGGTSolanum tuberosumCTAACAACCAAGCCGGTCCTGAATTCACTCCGly77AspGGAGTGAATTCAGGACCGGCTTGGTTGTTAGACCAGGAAGCCGC1350GGC-GACCACAACTTGAGCCACGTCAATATTGCTGCAATTCCTCCTAGCCTTAATGCTTAGCTGCCTCACGAAATTAGGAGGAACAATATTGACGTGGCTC1351GAGCCACGTCAATATTG1352Met OverproductionCTTCCTCTCTTATCCTTCGCTTTCCTCCCAACTTTGTCCGTCAGCT1353CGSCAGCACCAAGGCTCGCCACAACTGCAGCAACATTGGTGTCGCACMesembryanthemumAGGTCGTCGCTGCCTCCTGGTCCAACAACTCcrystallinumGAGTTGTTGGACCAGGAGGCAGCGACGACCTGTGCGACACCAAT1354Arg73HisGTTGCTGCAGTTGTGGCGAGCCTTGGTGCTGAGCTGACGGACAACGC-CACAGTTGGGAGGAAAGCGAAGGATAAGAGAGGAAGGGCTCGCCACAACTGCA1355TGCAGTTGTGGCGAGCC1356Met OverproductionTCCTTCGCTTTCCTCCCAACTTTGTCCGTCAGCTCAGCACCAAGG1357CGSCTCGCCGCAACTGCAACAACATTGGTGTCGCACAGGTCGTCGCTMesembryanthemumGCCTCCTGGTCCAACAACTCCGATGCCGGCGCcrystallinumGCGCCGGCATCGGAGTTGTTGGACCAGGAGGCAGCGACGACCT1358Ser77AsnGTGCGACACCAATGTTGTTGCAGTTGCGGCGAGCCTTGGTGCTGAGC-AACAGCTGACGGACAAAGTTGGGAGGAAAGCGAAGGACAACTGCAACAACATTG1359CAATGTTGTTGCAGTTG1360Met OverproductionTTTCCTCCCAACTTTGTCCGTCAGCTCAGCACCAAGGCTCGCCGC1361CGSAACTGCAGCAACATTAGTGTCGCACAGGTCGTCGCTGCCTCCTGMesembryanthemumGTCCAACAACTCCGATGCCGGCGCCACCTCTTcrystallinumAAGAGGTGGCGCCGGCATCGGAGTTGTTGGACCAGGAGGCAGC1362Gly80SerGACGACCTGTGCGACACTAATGTTGCTGCAGTTGCGGCGAGCCTGGT-AGTTGGTGCTGAGCTGACGGACAAAGTTGGGAGGAAAGCAACATTAGTGTCGCA1363TGCGACACTAATGTTGC1364Met OverproductionTTCCTCCCAACTTTGTCCGTCAGCTCAGCACCAAGGCTCGCCGCA1365CGSACTGCAGCAACATTGATGTCGCACAGGTCGTCGCTGCCTCCTGGTMesembryanthemumCCAACAACTCCGATGCCGGCGCCACCTCTTGcrystallinumCAAGAGGTGGCGCCGGCATCGGAGTTGTTGGACCAGGAGGCAG1366Gly80AspCGACGACCTGTGCGACATCAATGTTGCTGCAGTTGCGGCGAGCCGGT-GATTTGGTGCTGAGCTGACGGACAAAGTTGGGAGGAACAACATTGATGTCGCAC1367GTGCGACATCAATGTTG1368Met OverproductionCCTCTGCTACCATCCTCCGCTTTCCGCCAAACTTTGTCCGCCAGC1369CGSTTAGCACCAAGGCACACGGCAACTGCAGCAACATCGGCGTCGCGZea maysCAGATCGTCGCCGCCGCGTGGTCCGACTGCCCArg41HisGGGCAGTCGGACCACGCGGCGGCGACGATCTGCGCGACGCCGA1370CGC-CACTGTTGCTGCAGTTGCGGTGTGCCTTGGTGCTAAGCTGGCGGACAAAGTTTGGCGGAAAGCGGAGGATGGTAGCAGAGGCAAGGCACACCGCAACT1371AGTTGCGGTGTGCCTTG1372Met OverproductionTCCTCCGCTTTCCGCCAAACTTTGTCCGCCAGCTTAGCACCAAGG1373CGSCACGCCGCAACTGCAACAACATCGGCGTCGCGCAGATCGTCGCCZea maysGCCGCGTGGTCCGACTGCCCCGCCGCTCGCCCSer45AsnGGGCGAGCGGCGGGGCAGTCGGACCACGCGGCGGCGACGATCT1374AGC-AACGCGCGACGCCGATGTTGTTGCAGTTGCGGCGTGCCTTGGTGCTAAGCTGGCGGACAAAGTTTGGCGGAAAGCGGAGGACAACTGCAACAACATCG1375CGATGTTGTTGCAGTTG1376Met OverproductionTTTCCGCCAAACTTTGTCCGCCAGCTTAGCACCAAGGCACGCCGC1377CGSAACTGCAGCAACATCAGCGTCGCGCAGATCGTCGCCGCCGCGTGZea maysGTCCGACTGCCCCGCCGCTCGCCCCCACTTAGGly48SerCTAAGTGGGGGCGAGCGGCGGGGCAGTCGGACCACGCGGCGG1378GGC-AGCCGACGATCTGCGCGACGCTGATGTTGCTGCAGTTGCGGCGTGCCTTGGTGCTAAGCTGGCGGACAAAGTTTGGCGGAAAGCAACATCAGCGTCGCG1379CGCGACGCTGATGTTGC1380Met OverproductionTTCCGCCAAACTTTGTCCGCCAGCTTAGCACCAAGGCACGCCGCA1381CGSACTGCAGCAACATCGACGTCGCGCAGATCGTCGCCGCCGCGTGGZea maysTCCGACTGCCCCGCCGCTCGCCCCCACTTAGGGly48AspCCTAAGTGGGGGCGAGCGGCGGGGCAGTCGGACCACGCGGCG1382GGC-GACGCGACGATCTGCGCGACGTCGATGTTGCTGCAGTTGCGGCGTGCCTTGGTGCTAAGCTGGCGGACAAAGTTTGGCGGAACAACATCGACGTCGCGG1383GCGCGACGTCGATGTTG1384Met OverproductionGTATGAATGATCTGTGGGTGAAACACTGTGGGATTAGTCATACAG1385TSGAAGTTTCAAGGATCGTGGAATGACTGTTTTGGTTAGTCAAGTTAAArabidopsis thalianaTCGTCTGAGAAAGATGAAACGACCTGTGGTLeu205ArgACCACAGGTCGTTTCATCTTTCTCAGACGATTAACTTGACTAACCA1386CTT-CGTAAACAGTCATTCCACGATCCTTGAAACTTCCTGTATGACTAATCCCACAGTGTTTCACCCACAGATCATTCATACCAAGGATCGTGGAATGA1387TCATTCCACGATCCTTG1388Met OverproductionGCATGACTGATTTGTGGGTCAAACACTGTGGGATTAGCCATACTG1389TSGTAGTTTTAAGGATCGTGGGATGACTGTTTTGGTGAGTCAAGTTAASolanum tuberosumTCGCTTGCGGAAAATGCATAAACCGGTTGTLeu198ArgACAACCGGTTTATGCATTTTCCGCAAGCGATTAACTTGACTCACCA1390CTT-CGTAAACAGTCATCCCACGATCCTTAAAACTACCAGTATGGCTAATCCCACAGTGTTTGACCCACAAATCAGTCATGCTAAGGATCGTGGGATGA1391TCATCCCACGATCCTTA1392Lys OverproductionTCATTGGGCACACAGTGAACTGCTTTGGCTCTAGAATCAAAGTGA1393DHPSTAGGCAACACAGGAAACAACTCAACCAGAGAAGCCGTCCACGCAZea maysACAGAACAGGGATTTGCTGTTGGCATGCATGCSer157AsnGCATGCATGCCAACAGCAAATCCCTGTTCTGTTGCGTGGACGGCT1394AGC-AACTCTCTGGTTGAGTTGTTTCCTGTGTTGCCTATCACTTTGATTCTAGAGCCAAAGCAGTTCACTGTGTGCCCAATGACACAGGAAACAACTCAA1395TTGAGTTGTTTCCTGTG1396Lys OverproductionGCTCTAGAATCAAAGTGATAGGCAACACAGGAAGCAACTCAACCA1397DHPSGAGAAGCCGTCCACGAAACAGAACAGGGATTTGCTGTTGGCATGZea maysCATGCGGCTCTCCACATCAATCCTTACTACGGAla166ValCCGTAGTAAGGATTGATGTGGAGAGCCGCATGCATGCCAACAGC1398GCA-GAAAAATCCCTGTTCTGTTTCGTGGACGGCTTCTCTGGTTGAGTTGCTTCCTGTGTTGCCTATCACTTTGATTCTAGAGCCGTCCACGAAACAGAAC1399GTTCTGTTTCGTGGACG1400Lys OverproductionGGCTCTAGAATCAAAGTGATAGGCAACACAGGAAGCAACTCAACC1401DHPSAGAGAAGCCGTCCACACAACAGAACAGGGATTTGCTGTTGGCATZea maysGCATGCGGCTCTCCACATCAATCCTTACTACGAla166ThrCGTAGTAAGGATTGATGTGGAGAGCCGCATGCATGCCAACAGCA1402GCA-ACAAATCCCTGTTCTGTTGTGTGGACGGCTTCTCTGGTTGAGTTGCTTCCTGTGTTGCCTATCACTTTGATTCTAGAGCCCCGTCCACACAACAGAA1403TTCTGTTGTGTGGACGG1404Lys OverproductionTTATTGGGCATACAGTTAACTGCTTTGGCACTAAAATTAAAGTGGT1405DHPSCGGCAACACAGGAAATAACTCAACAAGGGAGGCTATTCACGCAACOryza sativaTGAGCAGGGATTCGCTGTAGGTATGCACGCSer24AsnGCGTGCATACCTACAGCGAATCCCTGCTCAGTTGCGTGAATAGCC1406AGT-AATTCCCTTGTTGAGTTATTTCCTGTGTTGCCGACCACTTTAATTTTAGTGCCAAAGCAGTTAACTGTATGCCCAATAACACAGGAAATAACTCAA1407TTGAGTTATTTCCTGTG1408Lys OverproductionGCACTAAAATTAAAGTGGTCGGCAACACAGGAAGTAACTCAACAA1409DHPSGGGAGGCTATTCACGTAACTGAGCAGGGATTCGCTGTAGGTATGOryza sativaCACGCGGCTCTCCACATCAATCCTTACTACGGAla133ValCCGTAGTAAGGATTGATGTGGAGAGCCGCGTGCATACCTACAGC1410GCA-GTAGAATCCCTGCTCAGTTACGTGAATAGCCTCCCTTGTTGAGTTACTTCCTGTGTTGCCGACCACTTTAATTTTAGTGCTATTCACGTAACTGAGC1411GCTCAGTTACGTGAATA1412Lys OverproductionGGCACTAAAATTAAAGTGGTCGGCAACACAGGAAGTAACTCAACA1413DHPSAGGGAGGCTATTCACACAACTGAGCAGGGATTCGCTGTAGGTATOryza sativaGCACGCGGCTCTCCACATCAATCCTTACTACGAla133ThrCGTAGTAAGGATTGATGTGGAGAGCCGCGTGCATACCTACAGCG1414GCA-ACAAATCCCTGCTCAGTTGTGTGAATAGCCTCCCTTGTTGAGTTACTTCCTGTGTTGCCGACCACTTTAATTTTAGTGCCCTATTCACACAACTGAG1415CTCAGTTGTGTGAATAG1416Lys OverproductionTCATCGGGCATACTGTTAACTGCTTTGGAGCCAACATTAAAGTGAT1417DHPS 1AGGCAACACGGGAAATAACTCAACCAGAGAAGCTGTTCACGCGATriticum aestivumCAGAGCAGGGATTTGCTGTTGGCATGCATGCSer65AsnGCATGCATGCCAACAGCAAATCCCTGCTCTGTCGCGTGAACAGCT1418AGT-AATTCTCTGGTTGAGTTATTTCCCGTGTTGCCTATCACTTTAATGTTGGCTCCAAAGCAGTTAACAGTATGCCCGATGACACGGGAAATAACTCAA1419TTGAGTTATTTCCCGTG1420Lys OverproductionGAGCCAACATTAAAGTGATAGGCAACACGGGAAGTAACTCAACCA1421DHPS 1GAGAAGCTGTTCACGTGACAGAGCAGGGATTTGCTGTTGGCATGTriticum aestivumCATGCAGCTCTTCATGTCAATCCTTACTACGGAla174ValCCGTAGTAAGGATTGACATGAAGAGCTGCATGCATGCCAACAGCA1422GCG-GTGAATCCCTGCTCTGTCACGTGAACAGCTTCTCTGGTTGAGTTACTTCCCGTGTTGCCTATCACTTTAATGTTGGCTCTGTTCACGTGACAGAGC1423GCTCTGTCACGTGAACA1424Lys OverproductionGGAGCCAACATTAAAGTGATAGGCAACACGGGAAGTAACTCAACC1425DHPS 1AGAGAAGCTGTTCACACGACAGAGCAGGGATTTGCTGTTGGCATTriticum aestivumGCATGCAGCTCTTCATGTCAATCCTTACTACGAla174ThrCGTAGTAAGGATTGACATGAAGAGCTGCATGCATGCCAACAGCAA1426GCG-ACGATCCCTGCTCTGTCGTGTGAACAGCTTCTCTGGTTGAGTTACTTCCCGTGTTGCCTATCACTTTAATGTTGGCTCCCTGTTCACAGGACAGAG1427CTCTGTCGTGTGAACAG1428Lys OverproductionTCATCGGGCACACTGTTAACTGCTTTGGAACTAACATTAAAGTGAT1429DHPS 2AGGCAACACGGGAAATAACTCAACTAGAGAAGCGATTCACGCTTCTriticum aestivumAGAGCAGGGATTTGCTGTTGGCATGCATGCSer154AsnGCATGCATGCCAACAGCAAATCCCTGCTCTGAAGCGTGAATCGCT1430AGT-AATTCTCTAGTTGAGTTATTTCCCGTGTTGCCTATCACTTTAATGTTAGTTCCAAAGCAGTTAACAGTGTGCCCGATGACACGGGAAATAACTCAA1431TTGAGTTATTTCCCGTG1432Lys OverproductionGAACTAACATTAAAGTGATAGGCAACACGGGAAGTAACTCAACTA1433DHPS 2GAGAAGCGATTCACGTTTCAGAGCAGGGATTTGCTGTTGGCATGCTriticum aestivumATGCAGCTCTCCATGTCAATCCTTACTATGGAla163ValCCATAGTAAGGATTGACATGGAGAGCTGCATGCATGCCAACAGCA1434GCT-GTTAATCCCTGCTCTGAAACGTGAATCGCTTCTCTAGTTGAGTTACTTCCCGTGTTGCCTATCACTTTAATGTTAGTTCGATTCACGTTTCAGAGC1435GCTCTGAAACGTGAATC1436Lys OverproductionGGAACTAACATTAAAGTGATAGGCAACACGGGAAGTAACTCAACT1437DHPS 2AGAGAAGCGATTCACACTTCAGAGCAGGGATTTGCTGTTGGCATGTriticum aestivumCATGCAGCTCTCCATGTCAATCCTTACTATGAla163ThrCATAGTAAGGATTGACATGGAGAGCTGCATGCATGCCAACAGCAA1438GCT-ACTATCCCTGCTCTGAAGTGTGAATCGCTTCTCTAGTTGAGTTACTTCCCGTGTTGCCTATCACTTTAATGTTAGTTCCCGATTCACACTTCAGAG1439CTCTGAAGTGTGAATCG1440Lys OverproductionCTCATTGGGCATACTGTGAACTGCTTTGGCTCTAGAATTAAAGTGA1441DHPSTAGGCAACACAGGAAATAACTCAACCAGAGAAGCTGTTCACGCAACoix lacryma-jobiCAGAGCAGGGATTTGCTGTTGGCATGCATGSer154AsnCATGCATGCCAACAGCAAATCCCTGCTCTGTTGCGTGAACAGCTT1442AGT-AATCTCTGGTTGAGTTATTTCCTGTGTTGCCTATCACTTTAATTCTAGAGCCAAAGCAGTTCACAGTATGCCCAATGAGCACAGGAAATAACTCAA1443TTGAGTTATTTCCTGTG1444Lys OverproductionGCTCTAGAATTAAAGTGATAGGCAACACAGGAAGTAACTCAACCA1445DHPSGAGAAGCTGTTCACGTAACAGAGCAGGGATTTGCTGTTGGCATGCCoix lacryma-jobiATGCAGCTCTCCACATCAATCCTTACTATGGAla163ValCCATAGTAAGGATTGATGTGGAGAGCTGCATGCATGCCAACAGCA1446GCA-GTAAATCCCTGCTCTGTTACGTGAACAGCTTCTCTGGTTGAGTTACTTCCTGTGTTGCCTATCACTTTAATTCTAGAGCTGTTCACGTAACAGAGC1447GCTCTGTTACGTGAACA1448Lys OverproductionGGCTCTAGAATTAAAGTGATAGGCAACACAGGAAGTAACTCAACC1449DHPSAGAGAAGCTGTTCACACAACAGAGCAGGGATTTGCTGTTGGCATGCoix lacryma-jobiCATGCAGCTCTCCACATCAATCCTTACTATGAla163ThrCATAGTAAGGATTGATGTGGAGAGCTGCATGCATGCCAACAGCAA1450GCA-ACAATCCCTGCTCTGTTGTGTGAACAGCTTCTCTGGTTGAGTTACTTCCTGTGTTGCCTATCACTTTAATTCTAGAGCCCTGTTCACACAACAGAG1451CTCTGTTGTGTGAACAG1452Lys OverproductionTCATTGGTCACACAGTCAATTGTTTTGGAGGGTCCATCAAAGTCAT1453DHPSCGGGAACACTGGAAACAACTCCACAAGGGAAGCAATCCATGCAANicotiana tabacumCTGAACAGGGATTTGCTGTAGGTATGCATGCSer136AsnGCATGCATACCTACAGCAAATCCCTGTTCAGTTGCATGGATTGCTT1454AGC-AACCCCTTGTGGAGTTGTTTCCAGTGTTCCCGATGACTTTGATGGACCCTCCAAAACAATTGACTGTGTGACCAATGACACTGGAAACAACTCCA1455TGGAGTTGTTTCCAGTG1456Lys OverproductionGAGGGTCCATCAAAGTCATCGGGAACACTGGAAGCAACTCCACAA1457DHPSGGGAAGCAATCCATGTAACTGAACAGGGATTTGCTGTAGGTATGCNicotiana tabacumATGCAGCTCTTCACATTAATCCCTACTATGGAla145ValCCATAGTAGGGATTAATGTGAAGAGCTGCATGCATACCTACAGCA1458GCA-GTAAATCCCTGTTCAGTTACATGGATTGCTTCCCTTGTGGAGTTGCTTCCAGTGTTCCCGATGACTTTGATGGACCCTCAATCCATGTAACTGAAC1459GTTCAGTTACATGGATT1460Lys OverproductionGGAGGGTCCATCAAAGTCATCGGGAACACTGGAAGCAACTCCAC1461DHPSAAGGGAAGCAATCCATACAACTGAACAGGGATTTGCTGTAGGTATNicotiana tabacumGCATGCAGCTCTTCACATTAATCCCTACTATGAla145ThrCATAGTAGGGATTAATGTGAAGAGCTGCATGCATACCTACAGCAA1462GCA-ACAATCCCTGTTCAGTTGTATGGATTGCTTCCCTTGTGGAGTTGCTTCCAGTGTTCCCGATGACTTTGATGGACCCTCCCAATCCATACAACTGAA1463TTCAGTTGTATGGATTG1464Lys OverproductionTTATAGGCCATACCGTTAACTGTTTTGGCGGAAGCATCAAAGTCAT1465DHPSTGGAAACACTGGAAACAATTCGACTAGAGAAGCAATCCACGCGACArabidopsis thalianaTGAACAAGGATTCGCGGTTGGAATGCATGCSer142AsnGCATGCATTCCAACCGCGAATCCTTGTTCAGTCGCGTGGATTGCT1466AGC-AACTCTCTAGTCGAATTGTTTCCAGTGTTTCCAATGACTTTGATGCTTCCGCCAAAACAGTTAACGGTATGGCCTATAACACTGGAAACAATTCGA1467TCGAATTGTTTCCAGTG1468Lys OverproductionGCGGAAGCATCAAAGTCATTGGAAACACTGGAAGCAATTCGACTA1469DHPSGAGAAGCAATCCACGTGACTGAACAAGGATTCGCGGTTGGAATGArabidopsis thalianaCATGCTGCTCTTCATATAAACCCTTACTATGGAla151ValCCATAGTAAGGGTTTATATGAAGAGCAGCATGCATTCCAACCGCG1470GCG-GTGAATCCTTGTTCAGTCACGTGGATTGCTTCTCTAGTCGAATTGCTTCCAGTGTTTCCAATGACTTTGATGCTTCCGCAATCCACGTGACTGAAC1471GTTCAGTCACGTGGATT1472Lys OverproductionGGCGGAAGCATCAAAGTCATTGGAAACACTGGAAGCAATTCGACT1473DHPSAGAGAAGCAATCCACACGACTGAACAAGGATTCGCGGTTGGAATArabidopsis thalianaGCATGCTGCTCTTCATATAAACCCTTACTATGAla151ThrCATAGTAAGGGTTTATATGAAGAGCAGCATGCATTCCAACCGCGA1474GCG-ACGATCCTTGTTCAGTCGTGTGGATTGCTTCTCTAGTCGAATTGCTTCCAGTGTTTCCAATGACTTTGATGCTTCCGCCCAATCCACACGACTGAA1475TTCAGTCGTGTGGATTG1476Lys OverproductionTTATTGCTCATACAGTCAACTGTTTTGGTGGGAAAATTAAGGTTAT1477DHPSTGGAAATACTGGAAACAACTCCACCAGGGAAGCAATTCATGCCACGlycine maxTGAGCAGGGTTTTGCTGTTGGAATGCATGCSer103AsnGCATGCATTCCAACAGCAAAACCCTGCTCAGTGGCATGAATTGCT1478AGC-AACTCCCTGGTGGAGTTGTTTCCAGTATTTCCAATAACCTTAATTTTCCCACCAAAACAGTTGACTGTATGAGCAATAATACTGGAAACAACTCCA1479TGGAGTTGTTTCCAGTA1480Lys OverproductionGTGGGAAAATTAAGGTTATTGGAAATACTGGAAGCAACTCCACCA1481DHPSGGGAAGCAATTCATGTCACTGAGCAGGGTTTTGCTGTTGGAATGCGlycine maxATGCTGCCCTTCACATAAACCCTTACTATGGAla112ValCCATAGTAAGGGTTTATGTGAAGGGCAGCATGCATTCCAACAGCA1482GCC-GTCAAACCCTGCTCAGTGACATGAATTGCTTCCCTGGTGGAGTTGCTTCCAGTATTTCCAATAACCTTAATTTTCCCACAATTCATGTCACTGAGC1483GCTCAGTGACATGAATT1484Lys OverproductionGGTGGGAAAATTAAGGTTATTGGAAATACTGGAAGCAACTCCACC1485DHPSAGGGAAGCAATTCATACCACTGAGCAGGGTTTTGCTGTTGGAATGGlycine maxCATGCTGCCCTTCACATAAACCCTTACTATGAla112ThrCATAGTAAGGGTTTATGTGAAGGGCAGCATGGATTCCAACAGCAA1486GCC-ACCAACCCTGCTCAGTGGTATGAATTGCTTCCCTGGTGGAGTTGCTTCCAGTATTTCCAATAACCTTAATTTTCCCACCCAATTCATACCACTGAG1487CTCAGTGGTATGAATTG1488Trp OverproductionCTTGCAGGAGACATATTTCAGATCGTGCTGAGTCAACGTTTTGAG1489ASCGGCGAACATTTGCAAACCCCTTTGAAGTTTATAGAGCACTAAGAArabidopsis thalianaGTTGTGAATCCAAGTCCGTATATGGGTTATTAsp341AsnAATAACCCATATACGGACTTGGATTCACAACTCTTAGTGCTCTATA1490GAG-AACAACTTCAAAGGGGTTTGCAAATGTTCGCCGCTCAAAACGTTGACTCAGCACGATCTGAAATATGTCTCCTGCAAGCATTTGCAAACCCCTTT1491AAAGGGGTTTGCAAATG1492Trp OverproductionGCTGCAGGAGACATATTTCAAATCGTTTTAAGTCAACGCTTTGAGA1493ASGAAGAACATTTGCTAACCCATTTGAAGTGTACAGAGCATTAAGAATNicotiana tabacumTGTGAATCCAAGCCCATATATGACTTACAAsp326AsnTGTAAGTCATATATGGGCTTGGATTCACAATTCTTAATGCTCTGTA1494GAC-AACCACTTCAAATGGGTTAGCAAATGTTCTTCTCTCAAAGCGTTGACTTAAAACGATTTGAAATATGTCTCCTGCAGCCATTTGCTAACCCATTT1495AAATGGGTTAGCAAATG1496Trp OverproductionCTAGCTGGTGACATTTTTCAAGTAGTCTTAAGCCAGCGTTTTGAGA1497ASGGCGTACATTTGCTAACCCCTTTGAGGTGTACCGTGCATTGCGTAOryza sativaTTGTCAATCCTAGTCCTTATATGGCCTATCAsp323AsnGATAGGCCATATAAGGACTAGGATTGACAATACGCAATGCACGGT1498GAC-AACACACCTCAAAGGGGTTAGCAAATGTACGCCTCTCAAAACGCTGGCTTAAGACTACTTGAAAAATGTCACCAGCTAGCATTTGCTAACCCCTTT1499AAAGGGGTTAGCAAATG1500Trp OverproductionCTTGCTGGTGACATATTCCAGATCGTACTAAGTCAGCGTTTTGAAA1501ASGGCGAACGTTCGCAAACCCATTTGAAATCTATAGATCACTGAGGARuta graveolensTTGTTAATCCAAGCCCATATATGACTTATTAsp354AsnAATAAGTCATATATGGGCTTGGATTAACAATCCTCAGTGATCTATA1502GAC-AACGATTTCAAATGGGTTTGCGAACGTTCGCCTTTCAAAACGCTGACTTAGTACGATCTGGAATATGTCACCAGCAAGCGTTCGCAAACCCATTT1503AAATGGGTTTGCGAACG1504Trp OverproductionCTGGCTGGGGACATATTCCAGCTTGTCCTAAGTCAGCGTTTTGAA1505ASCGGCGAACATTTGCAAATCCATTTGAAGTCTACCGAGCATTGAGACatharanthus roseusATTGTCAACCCAAGTCCATATATGACTTATTAsp354AsnAATAAGTCATATATGGACTTGGGTTGACAATTCTCAATGCTCGGTA1506GAT-AATGACTTCAAATGGATTTGCAAATGTTCGCCGTTCAAAACGCTGACTTAGGACAAGCTGGAATATGTCCCCAGCCAGCATTTGCAAATCCATTT1507AAATGGATTTGCAAATG1508

EXAMPLE 10

Production of Modified Starch in Plants

[0138] A principal aim of biotechnology is the improvement of crop plants for food value, agriculture, and to produce a range of plant-derived raw materials. Along with oils, fats and proteins, polysaccharides constitute the main raw materials derived from plants, and apart from cellulose, the storage polymer starch is the most important polysaccharide raw material. Starch is derived from a range of plants, but maize is the most important cultivated plant for the production of starch.

[0139] The polysaccharide starch is a polymer made up of glucose molecules. However, starch is not a homogeneous raw material and is, in fact, a highly complex mixture of various types of molecules which differ from each other, for example, in their degree of polymerization and in the degree of branching of the glucose chains. For example, amylose-starch is a basically non-branched polymer made up of α-1,4-glycosidically branched glucose molecules, and amylopectin-starch is a complex mixture of variously branched glucose chains. The branching results from additional α-1,6-glycosidic linkages. In plants from which starch is typically isolated, for example maize or potato, the starch is approximately 25% amylose-starch and 75% amylopectin-starch.

[0140] In maize, various mutants in starch metabolism are known, for example waxy, sugary, shrunken and opaque-2. In addition to producing a modified starch, these mutations greatly improve grain quality in maize, and thus expand the use of maize not only as the food but also for the important industrial materials in food chemistry. It would therefore be advantageous to be able readily to obtain mutants in these genes in particular maize genotypes as well as other plants. Such plants can be obtained, for example, using traditional breeding methods and through specific genetic modification by means of recombinant DNA techniques.

[0141] The attached tables disclose exemplary oligonucleotide base sequences which can be used to generate site-specific mutations in genes involved in starch metabolism.

22TABLE 20Genome-Altering Oligos Conferring Increased StarchPhenotype, Gene,Plant & TargetedSEQ IDAlterationAltering OligosNO:Increased StarchGAACTTGAGACTGAGAAAAGGGATCCAAGGACAGTTGCTTCCATT1509ADPGPPATTCTTGGAGGTGGAAAAGGAACTCGACTCTTTCCTCTCACAAAAArabidopsis thalianaCGCCGCGCCAAGCCTGCCGTTCCTATCGGGGAla99LysCCCCGATAGGAACGGCAGGCTTGGCGCGGCGTTTTGTGAGAGGA1510GCA-AAAAAGAGTCGAGTTCCTTTTCCACCTCCAAGAATAATGGAAGCAACTGTCCTTGGATCCCTTTTCTCAGTCTCAAGTTCGAGGTGGAAAAGGAACT1511AGTTCCTTTTCCACCTC1512Increased StarchCAAAACGCCGCGCCAAGCCTGCCGTTCCTATCGGGGGAGCCTAT1513ADPGPPAGGTTGATAGATGTACTAATGAGCAATTGTATTAACAGCGGAATCAArabidopsis thalianaACAAAGTCTACATACTCACACAATATAACTCPro127LeuGAGTTATATTGTGTGAGTATGTAGACTTTGTTGATTCCGCTGTTAA1514CCA-CTATACAATTGCTCATTAGTACATCTATCAACCTATAGGCTCCCCCGATAGGAACGGCAGGCTTGGCGCGGCGTTTTGAGATGTACTAATGAGCA1515TGCTCATTAGTACATCT1516Increased StarchTCACACAATATAACTCAGCATCATTGAACAGGCATTTAGCCCGTGC1517ADPGPPTTACAACTCCAATAATCTTGGCTTTGGAGATGGCTATGTTGAGGTTArabidopsis thalianaCTTGCGGCCACTCAAACGCCAGGAGAATCGly162AsnGATTCTCCTGGCGTTTGAGTGGCCGCAAGAACCTCAACATAGCCA1518GGA-AATTCTCCAAAGCCAAGATTATTGGAGTTGTAAGCACGGGGTAAATGCCTGTTCAATGATGCTGAGTTATATTGTGTGACTCCAATAATCTTGGCT1519AGCCAAGATTATTGGAG1520Increased StarchTCACACAATATAACTCAGCATCATTGAACAGGCATTTAGCCCGTGC1521ADPGPPTTACAACTCCAATAACCTTGGCTTTGGAGATGGCTATGTTGAGGTTArabidopsis thalianaCTTGCGGCCACTCAAACGCCAGGAGAATCGly162AsnGATTCTCCTGGCGTTTGAGTGGCCGCAAGAACCTCAACATAGCCA1522GGA-AACTCTCCAAAGCCAAGGTTATTGGAGTTGTAAGCACGGGCTAAATGCCTGTTCAATGATGCTGAGTTATATTGTGTGACTCCAATAACCTTGGCT1523AGCCAAGGTTATTGGAG1524Increased StarchGTTTGAGAGAAGAAAGGTAGACCCGCAAAATGTGGCTGCAATCAT1525ADPGPPTCTAGGAGGAGGCAAAGGAGCTAAACTCTTCCCTCTTACAATGAGArabidopsis thalianaAGCCGCAACACCAGCTGTAAATATTCATCTTAsn100LysAAGATGAATATTTACAGCTGGTGTTGCGGCTCTCATTGTAAGAGG1526AAT-AAAGAAGAGTTTAGCTCCTTTGCCTCCTCCTAGAATGATTGCAGCCACATTTTGCGGGTCTACCTTTCTTCTCTCAAACGGAGGCAAAGGAGCTAA1527TTAGCTCCTTTGCCTCC1528Increased StarchCTTGTGTCTTCAAATTATGTTAGGTTCCTGTTGGTGGATGCTACAG1529ADPGPPGCTGATCGATATCCTGATGAGTAACTGTATTAACAGCTGCATCAACArabidopsis thalianaAAGATATTTGTGCTGACACAGTTCAACTCPro128LeuGAGTTGAACTGTGTCAGCACAAATATCTTGTTGATGCAGCTGTTAA1530CCG-CTGTACAGTTACTCATCAGGATATCGATCAGCCTGTAGCATCCACCAACAGGAACCTAACATAATTTGAAGACACAAGCGATATCCTGATGAGTA1531TACTCATCAGGATATCG1532Increased StarchTGACACAGTTCAACTCAGCTTCCCTTAATCGACATTTAGCACGAAC1533ADPGPPTTATTTTGGGAATAATATAAACTTTGGAGGTGGTTTCGTAGAGGTAArabidopsis thalianaCAAACACTATGACAATAATAACTCTCAGCGly163AsnGCTGAGAGTTATTATTGTCATAGTGTTTGTACCTCTACGAAACCAC1534GGC-AATCTCCAAAGTTTATATTATTCCCAAAATAAGTTCGTGCTAAATGTCGATTAAGGGAAGCTGAGTTGAACTGTGTCATGGGAATAATATAAACT1535AGTTTATATTATTCCCA1536Increased StarchTGACACAGTTCAACTCAGCTTCCCTTAATCGACATTTAGCACGAAC1537ADPGPPTTATTTTGGGAATAACATAAACTTTGGAGGTGGTTTCGTAGAGGTAArabidopsis thalianaCAAACACTATGACAATAATAACTCTCAGCGly163AsnGCTGAGAGTTATTATTGTCATAGTGTTTGTACCTCTACGAAACCAC1538GGC-AACCTCCAAAGTTTATGTTATTCCCAAAATAAGTTCGTGCTAAATGTCGATTAAGGGAAGCTGAGTTGAACTGTGTCATGGGAATAACATAAACT1539AGTTTATGTTATTCCCA1540Increased StarchTTGAGGAACAACCAACGGCAGATCCAAAAGCTGTTGCCTCTGTCA1541ADPGPPTTCTAGGTGGTGGTAAAGGAACTCGTCTTTTTCCTCTTACAAGCALycopersiconGAAGAGCTAAACCAGCTGTTCCTATTGGTGGesculentumCCACCAATAGGAACAGCTGGTTTAGCTCTTCTGCTTGTAAGAGGA1542Val94LysAAAAGACGAGTTCCTTTACCACCACCTAGAATGACAGAGGCAACAGTT-AAAGCTTTTGGATCTGCCGTTGGTTGTTCCTCAATGGTGGTAAAGGAACTC1543GAGTTCCTTTACCACCA1544Increased StarchCAAGCAGAAGAGCTAAACCAGCTGTTCCTATTGGTGGTTGTTACC1545ADPGPPGGCTAATTGATGTACAAATGAGTAACTGCATTAACAGTGGCATACLycopersiconGGAAAATTTTCATCTTAACACAGTTCAATTCesculentumGAATTGAACTGTGTTAAGATGAAAATTTTCCGTATGCCACTGTTAA1546Pro122LeuTGCAGTTACTCATTTGTACATCAATTAGCCGGTAACAACCACCAATCCA-CAAAGGAACAGCTGGTTTAGCTCTTCTGGTTGTGATGTACAAATGAGTA1547TACTCATTTGTACATCA1548Increased StarchCACAGTTCAATTCCTTTTCCCTCAATCGTCACCTTGCCCGCACGTA1549ADPGPPTAATTTTGGAAATAATGTGGGTTTTGGAGATGGATTTGTGGAGGTTLycopersiconTTAGCTGCAACCCAGACTCCAGGGGATGCesculentumGCATCCCCTGGAGTCTGGGTTGCAGCTAAAACCTCCACAAATCCA1550Gly158AsnTCTCCAAAACCCACATTATTTCCAAAATTATACGTGCGGGCAAGGTGGA-AATGACGATTGAGGGAAAAGGAATTGAACTGTGTGGAAATAATGTGGGTT1551AACCCACATTATTTCCA1552Increased StarchCACAGTTCAATTCCTTTTCCCTCAATCGTCACCTTGCCCGCACGTA1553ADPGPPTAATTTTGGAAATAACGTGGGTTTTGGAGATGGATTTGTGGAGGTLycopersiconTTTAGCTGCAACCCAGACTCCAGGGGATGCesculentumGCATCCCCTGGAGTCTGGGTTGCAGCTAAAACCTCCACAAATCCA1554Gly158AsnTCTCCAAAACCCACGTTATTTCCAAAATTATACGTGCGGGCAAGGTGGA-AACGACGATTGAGGGAAAAGGAATTGAACTGTGTGGAAATAACGTGGGTT1555AACCCACGTTATTTCCA1556Increased StarchACGTAGATTTGGAAAAAAGAGACCCAAGTACAGTTGTAGCAATTAT1557ADPGPPACTAGGTGGAGGTAAAGGAACTCGTCTCTTCCCTCTCACCAAGCGCicer arietinumACGAGCCAAGCCTGCTGTTCCAATTGGAGGAla101LysCCTCCAATTGGAACAGCAGGCTTGGCTCGTCGCTTGGTGAGAGG1558GCT-AAAGAAGAGACGAGTTCCTTTACCTCCACCTAGTATAATTGCTACAACTGTACTTGGGTCTCTTTTTTCCAAATCTACGTTGGAGGTAAAGGAACTC1559GAGTTCCTTTACCTCCA1560Increased StarchCCAAGCGACGAGCCAAGCCTGCTGTTCCAATTGGAGGTGCTTATA1561ADPGPPGGCTGATAGATGTACTAATGAGTAACTGCATCAATAGTGGGATCACicer arietinumACAAAGTATACATTCTCACTCAATTTAATTCPro129LeuGAATTAAATTGAGTGAGAATGTATACTTTGTTGATCCCACTATTGA 1562CCA-CTATGCAGTTACTCATTAGTACATCTATCAGCCTATAAGCACCTCCAATTGGAACAGCAGGCTTGGCTCGTCGCTTGGAGATGTACTAATGAGTA1563TACTCATTAGTACATCT1564Increased StarchCTCAATTTAATTCAGCCTCACTCAACAGGCATATTGCACGTGCTTA1565ADPGPPTAACTCTGGTACTAATGTCACTTTTGGAGATGGCTATGTTGAGGTTCicer arietinumCTTGCAGCAACTCAAACTCCAGGGGAGCAGly165AsnTGCTCCCGTGGAGTTTGAGTTGCTGCAAGAACCTCAACATAGCCA1566GGA-AATTCTCCAAAAGTGACATTAGTACCAGAGTTATAAGCACGTGCAATATGCCTGTTGAGTGAGGCTGAATTAAATTGAGTGGTACTAATGTCACTT1567AAGTGACATTAGTACCA1568Increased StarchCTCAATTTAATTCAGCCTCACTCAACAGGCATATTGCACGTGCTTA1569ADPGPPTAACTCTGGTACTAACGTCACTTTTGGAGATGGCTATGTTGAGGTTCicer arietinumCTTGCAGCAACTCAAACTCCAGGGGAGCAGly165AsnTGCTCCCCTGGAGTTTGAGTTGCTGCAAGAACCTCAACATAGCCA1570GGA-AACTCTCCAAAAGTGACGTTAGTACCAGAGTTATAAGCACGTGCAATATGCCTGTTGAGTGAGGCTGAATTAAATTGAGTGGTACTAACGTCACTT1571AAGTGACGTTAGTACCA1572Increased StarchATATTGGAGAGGCGTCGGGCAAACCCTAAGAATGTGGCTGCAATC 1573ADPGPPATACTGCCAGGCGGTAAAGGGACACACCTATTCCCTCTCACCAATIpomoea batatasCGAGCTGCAACCCCTGCTGTTCCACTTGGAGAla94LysCTCCAAGTGGAACAGCAGGGGTTGCAGCTCGATTGGTGAGAGGG1574GCA-AAAAATAGGTGTGTCCCTTTACCGCCTGGCAGTATGATTGCAGCCACATTCTTAGGGTTTGCCCGACGCCTCTCCAATATCAGGCGGTAAAGGGACA1575TGTCCCTTTACCGCCTG1576Increased StarchCCAATCGAGCTGCAACCCCTGCTGTTCCACTTGGAGGATGCTATA1577ADPGPPGGTTGATCGACATTCTAATGAGCAACTGCATCAACAGCGGGGTTAIpomoea batatasACAAGATCTTTGTGCTGACCCAGTTCAATTCPro122LeuGAATTGAACTGGGTCAGCACAAAGATCTTGTTAACCCCGCTGTTG1578CCA-CTAATGCAGTTGCTCATTAGAATGTCGATCAACCTATAGCATCCTCCAAGTGGAACAGCAGGGGTTGCAGCTCGATTGGCGACATTCTAATGAGCA1579TGCTCATTAGAATGTCG1580Increased StarchTGACCCAGTTCAATTCAGCTTCTCTTAACCGTCACATTTCCCGTAC1581ADPGPPCGTCTTTGGCAATAATGTGAGCTTCGGAGATGGATTTGTTGAGGTIpomoea batatasGCTGGCTGCAACCCAAACACAAGGGGAAACGly157AsnGTTTCCCCTTGTGTTTGGGTTGCAGCCAGCACCTCAACAAATCCA1582GGT-AATTCTCCGAAGCTCACATTATTGCCAAAGACGGTACGGGAAATGTGACGGTTAAGAGAAGCTGAATTGAACTGGGTCATGGCAATAATGTGAGCT1583AGCTCACATTATTGCCA1584Increased StarchTGACCCAGTTCAATTCAGCTTCTCTTAACCGTCACATTTCCCGTAC1585ADPGPPCGTCTTTGGCAATAACGTGAGCTTCGGAGATGGATTTGTTGAGGTIpomoea batatasGCTGGCTGCAACCCAAACACAAGGGGAAACGly157AsnGTTTCCCCTTGTGTTTGGGTTGCAGCCAGCACCTCAACAAATCCA1586GGT-AACTCTCCGAAGCTCACGTTATTGCCAAAGACGGTACGGGAAATGTGACGGTTAAGAGAAGCTGAATTGAACTGGGTCATGGCAATAACGTGAGCT1587AGCTCACGTTATTGCCA1588Increased StarchCATTCCGGAGGAACTTTGCGGATCCAAATGAGGTTGCTGCTGTTA1589ADPGPPTATTGGGTGGTGGCAAAGGGACTCAACTTTTTCCTCTCACAAGCAOryza sativaCAAGGGCCACGCCTGCTGTTCCTATTGGAGGThr96LysCCTCCAATAGGAACAGCAGGCGTGGCCCTTGTGCTTGTGAGAGG1590ACC-AAAAAAAAGTTGAGTCCCTTTGCCACCACCCAATATAACAGCAGCAACCTCATTTGGATCCGCAAAGTTCCTCCGGAATGTGGTGGCAAAGGGACTC1591GAGTCCCTTTGCCACCA1592Increased StarchCAAGCACAAGGGCCACGCCTGCTGTTCCTATTGGAGGATGCTATA1593ADPGPPGGCTTATCGATATCCTCATGAGCAACTGTTTCAACAGTGGCATAAAOryza sativaCAAGATATTCATAATGACTCAATTCAACTCPro124LeuGAGTTGAATTGAGTCATTATGAATATCTTGTTTATGCCACTGTTGA1594CCC-CTCAACAGTTGCTCATGAGGATATCGATAAGCCTATAGCATCCTCCAATAGGAACAGCAGGCGTGGCCCTTGTGCTTGCGATATCCTCATGAGCA1595TGCTCATGAGGATATCG1596Increased StarchTGACTCAATTCAACTCAGCATCTCTTAATCGTCACATTCATCGTAC1597ADPGPPGTACCTTGGTGGTAATATCAACTTTACTGATGGTTCTGTTGAGGTAOryza sativaTTAGCCGCTACACAAATGCCTGGGGAGGCGly159AsnGCCTCCCCAGGCATTTGTGTAGCGGCTAATACCTCAACAGAACCA1598GGA-AATTCAGTAAAGTTGATATTACCACCAAGGTACGTACGATGAATGTGACGATTAAGAGATGCTGAGTTGAATTGAGTCATGGTGGTAATATCAACT1599AGTTGATATTACCACCA1600Increased StarchTGACTCAATTCAACTCAGCATCTCTTAATCGTCACATTCATCGTAC1601ADPGPPGTACCTTGGTGGTAACATCAACTTTACTGATGGTTCTGTTGAGGTAOryza sativaTTAGCCGCTACACAAATGCCTGGGGAGGCGly159AsnGCCTCCCCAGGCATTTGTGTAGCGGCTAATACCTCAACAGAACCA1602GGA-AACTCAGTAAAGTTGATGTTACCACCAAGGTACGTACGATGAATGTGACGATTAAGAGATGCTGAGTTGAATTGAGTCATGGTGGTAACATCAACT1603AGTTGATGTTACCACCA1604Increased StarchGTCCTTCAGGAGGATTAAGCGATCCGAACGAGGTTGCGGCCGTC1605ADPGPPATACTCGGCGGCGGCAAAGGGACTCAGCTCTTCCCACTCACGAGTriticum aestivumCACAAGGGCCACACCTGCTGTTCCTATTGGAGGThr80LysCCTCCAATAGGAACAGCAGGTGTGGCCCTTGTGCTCGTGAGTGG1606ACC-AAAGAAGAGCTGAGTCCCTTTGCCGCCGCCGAGTATGACGGCCGCAACCTCGTTCGGATCGCTTAATCCTCCTGAAGGACCGGCGGCAAAGGGACTC1607GAGTCCCTTTGCCGCCG1608Increased StarchCGAGCACAAGGGCCACACCTGCTGTTCCTATTGGAGGATGTTACA1609ADPGPPGGCTCATCGACATTCTCATGAGCAACTGCTTCAACAGTGGCATCATriticum aestivumACAAGATATTCGTCATGACCCAGTTCAACTCPro108LeuGAGTTGAACTGGGTCATGACGAATATCTTGTTGATGCCACTGTTG1610CCC-CTCAAGCAGTTGCTCATGAGAATGTCGATGAGCCTGTAACATCCTCCAATAGGAACAGCAGGTGTGGCCCTTGTGCTCGCGACATTCTCATGAGCA1611TGCTCATGAGAATGTCG1612Increased StarchTGACCCAGTTCAACTCGGCCTCCCTTAATCGTCACATTCACCGCA1613ADPGPPCCTACCTCGGCGGGAATATCAATTTCACTGATGGATCCGTTGAGGTriticum aestivumTATTGGCCGCGACGCAAATGCCCGGGGAGGCGly143AsnGCCTCCCCGGGCATTTGCGTCGCGGCCAATACCTCAACGGATCC1614GGA-AATATCAGTGAAATTGATATTCCCGCCGAGGTAGGTGCGGTGAATGTGACGATTAAGGGAGGCCGAGTTGAACTGGGTCACGGCGGGAATATCAATT1615AATTGATATTCCCGCCG1616Increased StarchTGACCCAGTTCAACTCGGCCTCCCTTAATCGTCACATTCACCGCA1617ADPGPPCCTACCTCGGCGGGAACATCAATTTCACTGATGGATCCGTTGAGGTriticum aestivumTATTGGCCGCGACGCAAATGCCCGGGGAGGCGly143AsnGCCTCCCCGGGCATTTGCGTCGCGGCCAATACCTCAACGGATCC1618GGA-AACATCAGTGAAATTGATGTTCCCGCCGAGGTAGGTGCGGTGAATGTGACGATTAAGGGAGGCCGAGTTGAACTGGGTCACGGCGGGAACATCAATT1619AATTGATGTTCCCGCCG1620Increased StarchCCTCCCGAAAGAATTATGCTGATGCAAGCCACGTTTCTGCTGTCA1621ADPGPPTTTTGGGTGGAGGCAAAGGAGTTCAACTCTTTCCTCTGACAAGCAOryza sativaCAAGGGCTACCCCCGCTGTTCCTGTTGGAGGThr95LysCCTCCAACAGGAACAGCGGGGGTAGCCCTTGTGCTTGTCAGAGG1622ACT-AAAAAAGAGTTGAACTCCTTTGCCTCCACCCAAAATGACAGCAGAAACGTGGCTTGCATCAGCATAATTCTTTCGGGAGGTGGAGGCAAAGGAGTTC1623GAACTCCTTTGCCTCCA1624Increased StarchCAAGCACAAGGGCTACCCCCGCTGTTCCTGTTGGAGGATGTTACA1625ADPGPPGGCTTATTGACATCCTTATGAGCAATTGCTTCAATAGCGGAATAAAOryza sativaTAAAATATTTGTGATGACTCAGTTCAATTCPro123LeuGAATTGAACTGAGTCATCACAAATATTTTATTTATTCCGCTATTGAA1626CCT-CTTGCAATTGCTCATAAGGATGTCAATAAGCCTGTAACATCCTCCAACAGGAACAGCGGGGGTAGCCCTTGTGCTTGTGACATCCTTATGAGCA1627TGCTCATAAGGATGTCA1628Increased StarchTGACTCAGTTCAATTCTGCTTCTCTTAATCGCCATATCCATCATACA1629ADPGPPTACCTTGGTGGGAATATCAACTTTACTGATGGGTCTGTGCAGGTAOryza sativaTTGGCTGCTACACAAATGCCTGACGAACCGly158AsnGGTTCGTCAGGCATTTGTGTAGCAGCCAATACCTGCACAGACCCA1630GGG-AATTCAGTAAAGTTGATATTCCCACCAAGGTATGTATGATGGATATGGCGATTAAGAGAAGCAGAATTGAACTGAGTCATGGTGGGAATATCAACT1631AGTTGATATTCCCACCA1632Increased StarchTGACTCAGTTCAATTCTGCTTCTCTTAATCGCCATATCCATCATACA1633ADPGPPTACCTTGGTGGGAACATCAACTTTACTGATGGGTCTGTGCAGGTAOryza sativaTTGGCTGCTACACAAATGCCTGACGAACCGly158AsnGGTTCGTCAGGCATTTGTGTAGCAGCCAATACCTGCACAGACCCA1634GGG-AACTCAGTAAAGTTGATGTTCCCACCAAGGTATGTATGATGGATATGGCGATTAAGAGAAGCAGAATTGAACTGAGTCATGGTGGGAACATCAACT1635AGTTGATGTTCCCACCA1636Increased StarchCCTTCCGCAGGAATTACGCCGATCCGAACGAGGTCGCGGCCGTC1637ADPGPPATACTCGGCGGTGGCAAAGGGACTCAGCTCTTCCCTCTCACAAGTriticum pestivumCACAAGGGCCACACCTGCTGTTCCTATTGGAGGThr99LysCCTCCAATAGGAACAGCAGGTGTGGCCCTTGTGCTTGTGAGAGG1638ACC-AAAGAAGAGCTGAGTCCCTTTGCCACCGCCGAGTATGACGGCCGCGACCTCGTTCGGATCGGCGTAATTCCTGCGGAAGGCGGTGGCAAAGGGACTC1639GAGTCCCTTTGCCACCG1640Increased StarchCAAGCACAAGGGCCACACCTGCTGTTCCTATTGGAGGATGTTACA1641ADPGPPGGCTCATCGATATTCTCATGAGCAACTGCTTCAATAGTGGCATCAATriticum aestivumCAAGATATTCGTCATGACGCAGTTCAACTCPro127LeuGAGTTGAACTGCGTCATGACGAATATCTTGTTGATGCCACTATTGA1642CCC-CTCAGCAGTTGCTCATGAGAATATCGATGAGCCTGTAACATCCTCCAATAGGAACAGCAGGTGTGGCCCTTGTGCTTGCGATATTCTCATGAGCA1643TGCTCATGAGAATATCG1644Increased StarchTGACGCAGTTCAACTCGGCCTCTCTTAATCGTCACATTCACCGCA1645ADPGPPCCTACCTCGGCGGGAATATCAATTTCACTGATGGATCTGTTGAGGTriticum aestivumTATTGGCCGCGACGCAAATGCCCGGGGAGGCGly162AsnGCCTCCCCGGGCATTTGCGTCGCGGCCAATACCTCAACAGATCC1646GGA-AATATCAGTGAAATTGATATTCCCGCCGAGGTAGGTGCGGTGAATGTGACGATTAAGAGAGGCCGAGTTGAACTGCGTCACGGCGGGAATATCAATT1647AATTGATATTCCCGCCG1648Increased StarchTGACGCAGTTCAACTCGGCCTCTCTTAATCGTCACATTCACCGCA1649ADPGPPCCTACCTCGGCGGGAACATCAATTTCACTGATGGATCTGTTGAGGTriticum aestivumTATTGGCCGCGACGCAAATGCCCGGGGAGGCGly162AsnGCCTCCCCGGGCATTTGCGTCGCGGCCAATACCTCAACAGATCC1650GGA-AACATCAGTGAAATTGATGTTCCCGCCGAGGTAGGTGCGGTGAATGTGACGATTAAGAGAGGCCGAGTTGAACTGCGTCACGGCGGGAACATCAATT1651AATTGATGTTCCCGCCG1652Increased StarchCTTTTCGGAGGAATTATGCTGATCCTAATGAAGTCGCTGCCGTCA1653ADPGPPTTTTGGGTGGTGGTAAAGGGACTCAGCTTTTCCCTCTCACAAGCAZea maysCAAGGGCCACCCCTGCTGTTCCTATTGGAGGThr96LysCCTCCAATAGGAACAGCAGGGGTGGCCCTTGTGCTTGTGAGAGG1654ACC-AAAGAAAAGCTGAGTCCCTTTACCACCACCCAAAATGACGGCAGCGAGTTCATTAGGATCAGCATAATTCCTCCGAAAAGTGGTGGTAAAGGGACTC1655GAGTCCCTTTACCACCA1656Increased StarchCAAGCACAAGGGCCACCCCTGCTGTTCCTATTGGAGGATGTTACA1657ADPGPPGGCTTATTGATATCCTCATGAGCAACTGTTTCAACAGTGGCATAAAZea maysCAAGATATTTGTTATGACTCAGTTCAACTCPro124LeuGAGTTGAACTGAGTCATAACAAATATCTTGTTTATGCCACTGTTGA1658CCC-CTCAACAGTTGCTCATGAGGATATCAATAAGCCTGTAACATCCTCCAATAGGAACAGCAGGGGTGGCCCTTGTGCTTGTGATATCCTCATGAGCA1659TGCTCATGAGGATATCA1660Increased StarchTGACTCAGTTCAACTCAGCTTCTCTTAACCGTCACATTCATCGTAC1661ADPGPPCTATCTTGGTGGGAATATCAACTTCACTGATGGATCTGTTGAGGTZea maysGCTGGCTGCAACACAAATGCCTGGGGAGGCGly159AsnGCCTCCCCAGGCATTTGTGTTGCAGCCAGCACCTCAACAGATCCA1662GGG-AATTCAGTGAAGTTGATATTCCCACCAAGATAGGTACGATGAATGTGACGGTTAAGAGAAGCTGAGTTGAACTGAGTCATGGTGGGAATATCAACT1663AGTTGATATTCCCACCA1664Increased StarchTGACTCAGTTCAACTCAGCTTCTCTTAACCGTCACATTCATCGTAC1665ADPGPPCTATCTTGGTGGGAACATCAACTTCACTGATGGATCTGTTGAGGTZea maysGCTGGCTGCAACACAAATGCCTGGGGAGGCGly159AsnGCCTCCCCAGGCATTTGTGTTGCAGCCAGCACCTCAACAGATCCA1666GGG-AACTCAGTGAAGTTGATGTTCCCACCAAGATAGGTACGATGAATGTGACGGTTAAGAGAAGCTGAGTTGAACTGAGTCATGGTGGGAACATCAACT1667AGTTGATGTTCCCACCA1668Increased StarchCTTGAGAGGCAAAAGAAGGGCGATGCAAGGACAGTAGTAGCAAT1669ADPGPPCATTCTAGGAGGGGGAAAGGGAACTCGTCTTTTCCCCCTCACCAASolanum tuberosumACGTCGTGCTAAGCCTGCCGTTCCAATGGGAGAla58LysCTCCCATTGGAACGGCAGGCTTAGCACGACGTTTGGTGAGGGGG1670GCG-AAGAAAAGACGAGTTCCCTTTCCCCCTCCTAGAATGATTGCTACTACTGTCCTTGCATCGCCCTTCTTTTGCCTCTCAAGGAGGGGGAAAGGGAACT1671AGTTCCCTTTCCCCCTC1672Increased StarchCCAAACGTCGTGCTAAGCCTGCCGTTCCAATGGGAGGAGCATATA1673ADPGPPGGCTAATTGATGTACTAATGAGCAACTGTATTAACAGTGGCATCAASolanum tuberosumCAAAGTATACATTCTCACTCAATTCAACTCPro86LeuGAGTTGAATTGAGTGAGAATGTATACTTTGTTGATGCCACTGTTAA1674CCA-CTATACAGTTGCTCATTAGTACATCAATTAGCCTATATGCTCCTCCCATTGGAACGGCAGGCTTAGCACGACGTTTGGTGATGTACTAATGAGCA1675TGCTCATTAGTACATCA1676Increased StarchCTCAATTCAACTCAGCCTCACTTAACAGGCATATAGCTCGTGCTTA1677ADPGPPCAACTTTGGCAATAATGTCACATTCGAGAGTGGCTATGTCGAGGTSolanum tuberosumCTTAGCAGCAACTCAAACACCAGGTGAATTGly122AsnAATTCACCTGGTGTTTGAGTTGCTGCTAAGACCTCGACATAGCCA1678GGG-AATCTCTCGAATGTGACATTATTGCCAAAGTTGTAAGCACGAGCTATATGCCTGTTAAGTGAGGCTGAGTTGAATTGAGTGGCAATAATGTCACAT1679ATGTGACATTATTGCCA1680Increased StarchCTCAATTCAACTCAGCCTCACTTAACAGGCATATAGCTCGTGCTTA1681ADPGPPCAACTTTGGCAATAACGTCACATTCGAGAGTGGCTATGTCGAGGTSolanum tuberosumCTTAGCAGCAACTCAAACACCAGGTGAATTGly122AsnAATTCACCTGGTGTTTGAGTTGCTGCTAAGACCTCGACATAGCCA1682GGG-AACCTCTCGAATGTGACGTTATTGCCAAAGTTGTAAGCACGAGCTATATGCCTGTTAAGTGAGGCTGAGTTGAATTGAGTGGCAATAACGTCACAT1683ATGTGACGTTATTGCCA1684Increased StarchTATTTGAATCTCCAAAAGCTGACCCAAAAAATGTGGCTGCAATTGT1685ADPGPPGCTGGGTGGTGGTAAAGGGACTCGCCTCTTTCCTCTTACTAGCAGBeta vulgarisGAGAGCTAAGCCAGCAGTGCCAATTGGAGGAla98LysCCTCCAATTGGCACTGCTGGCTTAGCTCTCCTGCTAGTAAGAGGA1686GCT-AAAAAGAGGCGAGTCCCTTTACCACCACCCAGCACAATTGCAGCCACATTTTTTGGGTCAGCTTTTGGAGATTCAAATATGGTGGTAAAGGGACTC1687GAGTCCCTTTACCACCA1688Increased StarchTATTTGAATCTCCAAAAGCTGACCCAAAAAATGTGGCTGCAATTGT1689ADPGPPGCTGGGTGGTGGTAACGGGACTCGCCTCTTTCCTCTTACTAGCAGBeta vulgarisGAGAGCTAAGCCAGCAGTGCCAATTGGAGGAla98LysCCTCCAATTGGCACTGCTGGCTTAGCTCTCCTGCTAGTAAGAGGA1690GCT-AACAAGAGGCGAGTCCCGTTACCACCACCCAGCACAATTGCAGCCACATTTTTTGGGTCAGCTTTTGGAGATTCAAATATGGTGGTAACGGGACTC1691GAGTCCCGTTACCACCA1692Increased StarchCTAGCAGGAGAGCTAAGCCAGCAGTGCCAATTGGAGGGTGTTAC1693ADPGPPAGGCTGATTGATGTGCTTATGAGCAACTGCATCAACAGTGGCATTBeta vulgarisAGAAAGATTTTCATTCTTACCCAGTTCAATTCPro126LeuGAATTGAACTGGGTAAGAATGAAAATCTTTCTAATGCCACTGTTGA1694CCT-CTTTGCAGTTGCTCATAAGCACATCAATCAGCCTGTAACACCCTCCAATTGGCACTGCTGGCTTAGCTCTCCTGCTAGTGATGTGCTTATGAGCA1695TGCTCATAAGCACATCA1696Increased StarchCCCAGTTCAATTCGTTTTCGCTTAATCGTCATCTTGCTCGAACCTA1697ADPGPPTAATTTTGGAGATAATGTGAATTTTGGGGATGGCTTTGTGGAGGTTBeta vulgarisTTTGCTGCTACACAAACACCTGGAGAATCGly162AsnGATTCTCCAGGTGTTTGTGTAGCAGCAAAAACCTCCACAAAGCCA1698GGT-AATTCCCCAAAATTCACATTATCTCCAAAATTATAGGTTCGAGCAAGATGACGATTAAGCGAAAACGAATTGAACTGGGTGGAGATAATGTGAATT1699AATTCACATTATCTCCA1700Increased StarchCCCAGTTCAATTCGTTTTCGCTTAATCGTCATCTTGCTCGAACCTA1701ADPGPPTAATTTTGGAGATAACGTGAATTTTGGGGATGGCTTTGTGGAGGTBeta vulgarisTTTTGCTGCTACACAAACACCTGGAGAATCGly162AsnGATTCTCCAGGTGTTTGTGTAGCAGCAAAAACCTCCACAAAGCCA1702GGT-AACTCCCCAAAATTCACGTTATCTCCAAAATTATAGGTTCGAGCAAGATGACGATTAAGCGAAAACGAATTGAACTGGGTGGAGATAACGTGAATT1703AATTCACGTTATCTCCA1704

[0142]

23

TABLE 21

Oligonucleotides to produce plants with waxy starch

Phenotype, Gene,

Plant & Targeted

SEQ ID

Alteration
Altering Oligos
NO:

Waxy starch
GAATCCAGGTAAACGGGTAGTTCATAATGGCAACTGTGACTGCTT
1705

GBSS
CTTCTAACTTTGTGTGAAGAACTTCACTTTTCAACAATCATGGTGCT

Arabidopsis thaliana

TCTTCATGCTCTGATGTCGCTCAGATTAC

Ser12Term
GTAATCTGAGCGACATCAGAGCATGAAGAAGCACCATGATTGTTG
1706

TCA-TGA
AAAAGTGAAGTTCTTCACACAAAGTTAGAAGAAGCAGTCACAGTTG

CCATTATGAACTACCCGTTTACCTGGATTC

CTTTGTGTGAAGAACTT
1707

AAGTTCTTCACACAAAG
1708

Waxy starch
ATCCAGGTAAACGGGTAGTTCATAATGGCAACTGTGACTGCTTCTT
1709

GBSS
CTAACTTTGTGTCATGAACTTCACTTTTCAACAATCATGGTGCTTCT

Arabidopsis thaliana

TCATGCTCTGATGTCGCTCAGATTACCT

Arg13Term
AGGTAATCTGAGCGACATCAGAGCATGAAGAAGCACCATGATTGT
1710

AGA-TGA
TGAAAAGTGAAGTTCATGACACAAAGTTAGAAGAAGCAGTCACAGT

TGCCATTATGAACTACCCGTTTACCTGGAT

TTGTGTCATGAACTTCA
1711

TGAAGTTCATGACACAA
1712

Waxy starch
TAAACGGGTAGTTCATAATGGCAACTGTGACTGCTTCTTCTAACTT
1713

GBSS
TGTGTCAAGAACTTGACTTTTCAACAATCATGGTGCTTCTTCATGCT

Arabidopsis thaliana

CTGATGTCGCTCAGATTACCTTAAAAGG

Ser15Term
CCTTTTAAGGTAATCTGAGCGACATCAGAGCATGAAGAAGCACCAT
1714

TCA-TGA
GATTGTTGAAAAGTCAAGTTCTTGACACAAAGTTAGAAGAAGCAGT

CACAGTTGCCATTATGAACTACCCGTTTA

AAGAACTTGACTTTTCA
1715

TGAAAAGTCAAGTTCTT
1716

Waxy starch
TGACTGCTTCTTCTAACTTTGTGTCAAGAACTTGACTTTTCAACAAT
1717

GBSS
CATGGTGCTTCTTGATGCTCTGATGTCGCTCAGATTACCTTAAAAG

Arabidopsis thaliana

GCCAATCCTTGACTCATTGTGGGTTAAG

Ser24Term
CTTAACCCACAATGAGTCAAGGATTGGCCTTTTAAGGTAATCTGAG
1718

TCA-TGA
CGACATCAGAGCATCAAGAAGCACCATGATTGTTGAAAAGTGAAG

TTCTTGACACAAAGTTAGAAGAAGCAGTCA

TGCTTCTTGATGCTCTG
1719

CAGAGCATCAAGAAGCA
1720

Waxy starch
TGCTTCTTCTAACTTTGTGTCAAGAACTTCACTTTTCAACAATCATG
1721

GBSS
GTGCTTCTTCATGATCTGATGTCGCTCAGATTACCTTAAAAGGCCA

Arabidopsis thaliana

ATCCTTGACTCATTGTGGGTTAAGGTCA

Cys25Term
TGACCTTAACCCACAATGAGTCAAGGATTGGCCTTTTAAGGTAATC
1722

TGC-TGA
TGAGCGACATCAGATCATGAAGAAGCACCATGATTGTTGAAAAGT

GAAGTTCTTGACACAAAGTTAGAAGAAGCA

TCTTCATGATCTGATGT
1723

ACATCAGATCATGAAGA
1724

Waxy starch
GTAACAGCTTCACAGTTGGTGTCACATGTCCATGGTGGAGCAACG
1725

GBSS
TCTTCACCGGATACTTAAACAAACTTGGCCCAGGTTGGCCTCAGG

Antirrhinum majus

AACCAGCAATTCACTCACAATGGGTTGAGAT

Lys24Term
ATCTCAAGCCATTGTGAGTGAATTGCTGGTTCGTGAGGCCAACCTG
1726

AAA-TAA
GGCCAAGTTTGTTTAAGTATCGGGTGAAGACGTTGCTCCACCATG

GACATGTGACACCAACTGTGAAGGTGTTAC

CGGATACTTAAACAAAC
1727

GTTTGTTTAAGTATCCG
1728

Waxy starch
CACAGTTGGTGTCACATGTCCATGGTGGAGCAAGGTCTTCACCGG
1729

GBSS
ATAGTAAAACAAACTAGGGCGAGGTTGGCCTCAGGAACCAGCAAT

Antirrhinum majus

TCACTCACAATGGGTTGAGATCAATAAACAT

Leu27Term
ATGTTTATTGATCTCAACCCATTGTGAGTGAATTGCTGGTTCCTGA
1730

TTG-TAG
GGCCAACCTGGGCCTAGTTTGTTTTAGTATCGGGTGAAGACGTTG

CTCCACCATGGACATGTGACACCAACTGTG

AACAAACTAGGCCCAGG
1731

CCTGGGCCTAGTTTGTT
1732

Waxy starch
TTGGTGTCACATGTCCATGGTGGAGCAACGTCTTCACCGGATACT
1733

GBSS
AAAACAAACTTGGCCTAGGTTGGCCTCAGGAACCAGCAATTCACT

Antirrhinum majus

CACAATGGGTTGAGATCAATAAACATGGTTG

Gln29Term
CAACCATGTTTATTGATCTCAACCCATTGTGAGTGAATTGCTGGTT
1734

GAG-TAG
CCTGAGGCCAACCTAGGCCAAGTTTGTTTTAGTATCCGGTGAAGA

CGTTGCTCCACCATGGACATGTGACACCAA

ACTTGGCCTAGGTTGGC
1735

GCCAACCTAGGCCAAGT
1736

Waxy starch
GGTGGAGCAACGTCTTCACCGGATACTAAAACAAACTTGGCCCAG
1737

GBSS
GTTGGCCTCAGGAACTAGCAATTCACTCACAATGGGTTGAGATCA

Antirrhinum majus

ATAAACATGGTTGATAAGCTTCAAATGAGGA

Gln35Term
TCCTCATTTGAAGCTTATCAACCATGTTTATTGATGTCAACCCATTG
1738

GAG-TAG
TGAGTGAATTGCTAGTTCCTGAGGCCAACCTGGGCCAAGTTTGTTT

TAGTATCCGGTGAAGACGTTGCTCCACC

TCAGGAACTAGCAATTC
1739

GAATTGCTAGTTCCTGA
1740

Waxy starch
GGAGCAACGTCTTCACCGGATACTAAAACAAACTTGGCCCAGGTT
1741

GBSS
GGCCTCAGGAACCAGTAATTCACTCACAATGGGTTGAGATCAATAA

Antirrhinum majus

ACATGGTTGATAAGCTTCAAATGAGGAACA

Gln36Term
TGTTCCTCATTTGAAGCTTATCAACCATGTTTATTGATCTCAACCCA
1742

CAA-TAA
TTGTGAGTGAATTACTGGTTCCTGAGGCCAACCTGGGCCAAGTTT

GTTTTAGTATCCGGTGAAGACGTTGCTCC

GGAACCAGTAATTCACT
1743

AGTGAATTACTGGTTCC
1744

Waxy starch
GTGATGGCGACTATAACTGCCTCACACTTTGTTTCTCATGTCTGTG
1745

GBSS
GGGGTGCCACTTCTTGAGAATCAAAAGTGGGGTTGGGTCAATTAG

Ipomoea batatas

CCCTGAGGAGCCAAGCTGTGACTCACAATG

Gly20Term
CATTGTGAGTCACAGCTTGGCTCCTCAGGGCTAATTGACCCAACC
1746

GGA-TGA
CCACTTTTGATTCTCAAGAAGTGGCACCCCCACAGACATGAGAAA

CAAAGTGTGAGGCAGTTATAGTCGCCATCAC

CCACTTCTTGAGAATCA
1747

TGATTCTCAAGAAGTGG
1748

Waxy starch
ATGGCGACTATAACTGCCTCACACTTTGTTTCTCATGTCTGTGGGG
1749

GBSS
GTGCCACTTCTGGATAATCAAAAGTGGGGTTGGGTCAATTAGCCC

Ipomoea batatas

TGAGGAGCCAAGCTGTGACTCACAATGGGT

Glu21Term
ACCCATTGTGAGTCACAGCTTGGCTCCTCAGGGCTAATTGACCCA
1750

GAA-TAA
ACCCCACTTTTGATTATCCAGAAGTGGCACCCCCACAGACATGAG

AAACAAAGTGTGAGGCAGTTATAGTCGCCAT

CTTCTGGATAATCAAAA
1751

TTTTGATTATCCAGAAG
1752

Waxy starch
CGACTATAACTGCCTCACACTTTGTTTCTCATGTCTGTGGGGGTGC
1753

GBSS
CACTTCTGGAGAATGAAAAGTGGGGTTGGGTCAATTAGCCCTGAG

Ipomoea batatas

GAGCCAAGCTGTGACTCACAATGGGTTGAG

Ser22Term
CTCAACCCATTGTGAGTCACAGCTTGGCTCCTCAGGGCTAATTGA
1754

TCA-TGA
CCCAACCCCACTTTTCATTCTCCAGAAGTGGCACCCCCACAGACAT

GAGAAACAAAGTGTGAGGCAGTTATAGTCG

TGGAGAATGAAAAGTGG
1755

CCACTTTTCATTCTCCA
1756

Waxy starch
ACTATAACTGCCTCACACTTTGTTTCTCATGTCTGTGGGGGTGCCA
1757

GBSS
CTTCTGGAGAATCATAAGTGGGGTTGGGTCAATTAGCCCTGAGGA

Ipomoea batatas

GCCAAGCTGTGACTCACAATGGGTTGAGAC

Lys23Term
GTCTCAACCCATTGTGAGTCACAGCTTGGCTCCTCAGGGCTAATT
1758

AAA-TAA
GACCCAACCCCACTTATGATTCTCCAGAAGTGGCACCCCCACAGA

CATGAGAAACAAAGTGTGAGGCAGTTATAGT

GAGAATCATAAGTGGGG
1759

CCCCACTTATGATTCTC
1760

Waxy starch
CCTCACACTTTGTTTCTCATGTCTGTGGGGGTGCCACTTCTGGAGA
1761

G BSS
ATCAAAAGTGGGGTAGGGTCAATTAGCCCTGAGGAGCCAAGCTGT

Ipomoea batatas

GACTCACAATGGGTTGAGACCTGTGAACAA

Leu26Term
TTGTTCACAGGTCTCAACCCATTGTGAGTCACAGCTTGGCTCCTCA
1762

TTG-TAG
GGGCTAATTGACCCTACCCCACTTTTGATTCTCCAGAAGTGGCACC

CCCACAGACATGAGAAACAAAGTGTGAGG

AGTGGGGTAGGGTCAAT
1763

ATTGACCCTACCCCACT
1764

Waxy starch
CATCGGCGATTGTTGCTCCTTACTGCTCTCTCACAGAATGGCAACG
1765

GBSS
GTGACGGGGTCTTAGGTGGTGTCGAGAAGCGCGTGCTTCAATTCC

Astragalus

CAGGGAAGAACAGAAGCCAAAGTGAATTCA

membranaeus

TGAATTCACTTTGGCTTCTGTTCTTCCCTGGGAATTGAAGCACGCG
1766

Tyr8Term
CTTCTCGACACCACCTAAGACCCCGTCACCGTTGCCATTCTGTGA

TAT-TAG
GAGAGCAGTAAGGAGCAACAATCGCCGATG

GGGTCTTAGGTGGTGTC
1767

GACACCACCTAAGACCC
1768

Waxy starch
ATTGTTGCTCCTTACTGCTCTCTCACAGAATGGCAACGGTGACGG
1769

GBSS
GGTCTTATGTGGTGTAGAGAAGCGCGTGCTTCAATTCCCAGGGAA

Astragalus

GAACAGAAGCCAAAGTGAATTCACCTCAGAA

membranaeus

TTCTGAGGTGAATTCACTTTGGCTTCTGTTCTTCCCTGGGAATTGA
1770

Ser11Term
AGCACGCGCTTCTCTACACCACATAAGACCCCGTCACCGTTGCCA

TCG-TAG
TTCTGTGAGAGAGCAGTAAGGAGCAACAAT

TGTGGTGTAGAGAAGCG
1771

CGCTTCTCTACACCACA
1772

Waxy starch
TGTTGCTCCTTACTGCTCTCTCACAGAATGGCAACGGTGACGGGG
1773

GBSS
TCTTATGTGGTGTCGTGAAGCGCGTGCTTCAATTCCCAGGGAAGA

Astragalus

ACAGAAGCCAAAGTGAATTCACCTCAGAAGA

membranaeus

TCTTCTGAGGTGAATTCACTTTGGCTTCTGTTCTTCCCTGGGAATT
1774

Arg12Term
GAAGCACGCGCTTCACGACACCACATAAGACCCCGTCACCGTTGC

AGA-TGA
CATTCTGTGAGAGAGCAGTCAGGAGCAACA

TGGTGTCGTGAAGCGCG
1775

CGCGCTTCACGACACCA
1776

Waxy starch
ACTGCTCTCTCACAGAATGGCAACGGTGACGGGGTCTTATGTGGT
1777

GBSS
GTCGAGAAGCGCGTGATTCAATTCCCAGGGAAGAACAGAAGCCAA

Astragalus

AGTGAATTCACCTCAGAAGATAAATCTGAAT

membranaeus

ATTGAGATTTATCTTCTGAGGTGAATTCACTTTGGCTTCTGTTCTTC
1778

Cys15Term
CCTGGGAATTGAATCACGCGCTTCTCGACACCACATAAGACCCCG

TGC-TGA
TCACCGTTGCCATTCTGTGAGAGAGCAGT

AGCGCGTGATTCAATTC
1779

GAATTGAATCACGCGCT
1780

Waxy starch
CACAGAATGGCAACGGTGACGGGGTCTTATGTGGTGTCGAGAAG
1781

GBSS
CGCGTGGTTCAATTCCTAGGGAAGAACAGAAGCCAAAGTGAATTC

Astragalus

ACCTCAGAAGATAAATCTCAATAGCCAAGCAT

membranaeus

ATGCTTGGCTATTGAGATTTATCTTCTGAGGTGAATTCACTTTGGCT
1782

Gln19Term
TCTGTTCTTCCCTAGGAATTGAAGCACGCGCTTCTCGACACCACAT

CAG-TAG
AAGACCCCGTCACCGTTGCCATTCTGTG

TCAATTCCTAGGGAAGA
1783

TCTTCCCTAGGAATTGA
1784

Waxy starch
TGTAGCTTGGTAGATTCCCCTTTTTGTCGACCACACATCACATGGC
1785

GBSS
AAGCATCACAGCTTGACACCACTTTGTGTCAAGAAGCCAAACTTCA

Solanum tuberosum

CTAGACACCAAATCAACCTTGTCACAGAT

Ser7Term
ATCTGTGACAAGGTTGATTTGGTGTCTAGTGAAGTTTGGCTTCTTG
1786

TCA-TGA
ACACAAAGTGGTGTCAAGCTGTGATGCTTGCCATGTGATGTGTGG

TCTACAAAAAGGGGAATCTACCAAGCTACA

CACAGCTTGACACCACT
1787

AGTGGTGTCAAGCTGTG
1788

Waxy starch
TCCCCTTTTTGTAGACCACACATCACATGGCAAGCATCACAGCTTC
1789

GBSS
ACACCACTTTGTGTGAAGAAGCCAAACTTCACTAGACACCAAATCA

Solanum tuberosum

ACCTTGTCACAGATAGGACTCAGGAACCA

Ser12Term
TGGTTCCTGAGTCCTATCTGTGACAAGGTTGATTTGGTGTCTAGTG
1790

TCA-TGA
AAGTTTGGCTTCTTCACACAAAGTGGTGTGAAGCTGTGATGCTTGC

CATGTGATGTGTGGTCTACAAAAAGGGGA

CTTTGTGTGAAGAAGCC
1791

GGCTTCTTCACACAAAG
1792

Waxy starch
CCCTTTTTGTAGACCACACATCACATGGCAAGCATCACAGCTTCAC
1793

GBSS
ACCACTTTGTGTCATGAAGCCAAACTTCACTAGACACCAAATCAAC

Solanum tuberosum

CTTGTCACAGATAGGACTCAGGAACCATA

Arg13Term
TATGGTTCCTGAGTCCTATCTGTGACAAGGTTGATTTGGTGTCTAG
1794

AGA-TGA
TGAAGTTTGGCTTCATGACACAAAGTGGTGTGAAGCTGTGATGCTT

GCCATGTGATGTGTGGTCTACAAAAAGGG

TTGTGTCATGAAGCCAA
1795

TTGGCTTCATGACACAA
1796

Waxy starch
TTGTAGACCACACATCACATGGCAAGCATCACAGCTTCACACCACT
1797

GBSS
TTGTGTCAAGAAGCTAAACTTCACTAGACACCAAATCAACCTTGTC

Solanum tuberosum

ACAGATAGGACTCAGGAACCATACTCTGA

Gln15Term
TCAGAGTATGGTTCCTGAGTCCTATCTGTGACAAGGTTGATTTGGT
1798

CAA-TAA
GTCTAGTGAAGTTTAGCTTCTTGACACAAAGTGGTGTGAAGCTGTG

ATGCTTGCCATGTGATGTGTGGTCTACAA

CAAGAAGCTAAACTTCA
1799

TGAAGTTTAGCTTCTTG
1800

Waxy starch
CCACACATCACATGGCAAGCATCACAGCTTCACACCACTTTGTGTC
1801

GBSS
AAGAAGCCAAACTTGACTAGACACCAAATCAACCTTGTCACAGATA

Solanum tuberosum

GGACTCAGGAACCATACTCTGACTCACAA

Sen17Term
TTGTGAGTCAGAGTCTGGTTCCTGAGTCCTATCTGTGACAAGGTTG
1802

TCA-TGA
ATTTGGTGTCTAGTCAAGTTTGGCTTGTTGACACAAAGTGGTGTGA

AGCTGTGATGCTTGCCATGTGATGTGTGG

CCAAACTTGACTAGACA
1803

TGTCTAGTCAAGTTTGG
1804

Waxy starch
GTCGATCACTCTTCTCTCACCGCCGAAACAGATTTTGACACAAAAA
1805

GBSS
TGGCAACAATAACGTGATCTTCAATGCCGACGAGAACCGCGTGCT

Pisum sativum

TCAATTACCAAGGAAGATCAGCAGAGTCTA

Gly6Term
TAGACTCTGCTGATCTTCCTTGGTCATTGAAGCACGCGGTTCTCGT
1806

GGA-TGA
CGGCATTGAAGATCACGTTATTGTTGCCATTTTTGTGTCAAAATCT

GTTTCGGCGGTGAGAGAAGAGTGATCGAC

CAATAACGTGATCTTCA
1807

TGAAGATCACGTTATTG
1808

Waxy starch
ACTCTTCTCTCACCGCCGAAACAGATTTTGACACAAAAATGGCAAC
1809

GBSS
AATAACGGGATCTTGAATGCCGACGAGAACCGCGTGCTTCAATTA

Pisum sativum

CCAAGGAAGATCAGCAGAGTCTAAACTGAA

Ser8Term
TTCAGTTTAGACTCTGCTGATCTTCCTTGGTCATTGAAGCACGCGG
1810

TCA-TGA
TTCTCGTCGGCATTCAAGATCCCGTTATTGTTGCCATTTTTGTGTCA

AAATCTGTTTCGGCGGTGAGAGAAGAGT

GGGATCTTGAATGCCGA
1811

TCGGCATTCAAGATCCC
1812

Waxy starch
ACCGCCGAAACAGATTTTGACACAAAAATGGCAACAATAACGGGA
1813

GBSS
TCTTCAATGCCGACGTGAACCGCGTGCTTCAATTACCAAGGAAGA

Pisum sativum

TCAGCAGAGTCTAAACTGAATTTGCCTCAGA

Arg12Term
TCTGAGGCAAATTCAGTTTAGACTCTGCTGATCTTCCTTGGTCATT
1814

AGA-TGA
GAAGCACGCGGTTCACGTCGGCATTGAAGATCCCGTTATTGTTGC

CATTTTTGTGTCAAAATCTGTTTCGGCGGT

TGCCGACGTGAACCGCG
1815

CGCGGTTCACGTCGGCA
1816

Waxy starch
AGATTTTGACACAAAAATGGCAACAATAACGGGATCTTCAATGCCG
1817

GBSS
ACGAGAACCGCGTGATTCAATTACCAAGGAAGATCAGCAGAGTCT

Pisum sativum

AAACTGAATTTGCCTCAGATACACTTCAAT

Cys15Term
ATTGAAGTGTCTCTGAGGCAAATTCAGTTTAGACTCTGCTGATCTT
1818

TGC-TGA
CCTTGGTCATTGAATCACGCGGTTCTCGTCGGCATTGAAGATCCC

GTTATTGTTGCCATTTTTGTGTCAAAATCT

ACCGCGTGATTCAATTA
1819

TAATTGAATCACGCGGT
1820

Waxy starch
CACAAAAATGGCAACAATAACGGGATCTTCAATGCCGACGAGAAC
1821

GBSS
CGCGTGCTTCAATTAGCAAGGAAGATCAGCAGAGTCTAAACTGAA

Pisum sativum

TTTGCCTCAGATACACTTCAATAACAACCAA

Tyr18Term
TTGGTTGTTATTGAAGTGTATCTGAGGCAAATTCAGTTTAGACTCT
1822

TAC-TAG
GCTGATCTTCCTTGCTAATTGAAGCACGCGGTTCTCGTCGGCATTG

AAGATCCCGTTATTGTTGCCATTTTTGTG

TTCAATTAGCAAGGAAG
1823

CTTCCTTGCTAATTGAA
1824

Waxy starch
TCTACACCGGAGAGAGCACCATGGCAACTGTAATAGCTGCACATT
1825

GBSS
TCGTTTCCAGGAGCTGACACTTGAGCATCCATGCATTAGAGACTAA

Manihot esculenta

GGCTAATAATTTGTCTCACACTGGACCCTG

Ser14Term
CAGGGTCCAGTGTGAGACAAATTATTAGCCTTAGTCTCTAATGCAT
1826

TCA-TGA
GGATGCTCAAGTGTCAGCTCCTGGAAACGAAATGTGCAGCTATTA

CAGTTGCCATGGTGCTCTCTCCGGTGTAGA

CAGGAGCTGACACTTGA
1827

TCAAGTGTCAGCTCCTG
1828

Waxy starch
CCGGAGAGAGCACCATGGCAACTGTAATAGCTGCACATTTCGTTT
1829

GBSS
CCAGGAGCTCACACTAGAGCATCCATGCATTAGAGACTAAGGCTA

Manihot esculenta

ATAATTTGTCTCACACTGGACCCTGGACCCA

Leu16Term
TGGGTCCAGGGTCCAGTGTGAGACAAATTATTAGCCTTAGTCTCTA
1830

TTG-TAG
ATGCATGGATGCTCTAGTGTGAGCTCCTGGAAACGAAATGTGCAG

CTATTACAGTTGCCATGGTGCTCTCTCCGG

CTCACACTAGAGCATCC
1831

GGATGCTCTAGTGTGAG
1832

Waxy starch
TGGCAACTGTAATAGCTGCACATTTCGTTTCCAGGAGCTCACACTT
1833

GBSS
GAGCATCCATGCATGAGAGACTAAGGCTAATAATTTGTCTCACACT

Manihot esculenta

GGACCCTGGACCCAAACTATCACTCCCAA

Leu21Term
TTGGGAGTGATAGTTTGGGTCCAGGGTCCAGTGTGAGACAAATTA
1834

TTA-TGA
TTAGCCTTAGTCTCTCATGCATGGATGCTCAAGTGTGAGCTCCTGG

AAACGAAATGTGCAGCTATTACAGTTGCCA

CCATGCATGAGAGACTA
1835

TAGTCTCTCATGCATGG
1836

Waxy starch
GCAACTGTAATAGCTGCACATTTCGTTTCCAGGAGCTCACACTTGA
1837

GBSS
GCATCCATGCATTATAGACTAAGGCTAATAATTTGTCTCACACTGG

Manihot esculenta

ACCCTGGACCCAAACTATCACTCCCAATG

Glu22Term
CATTGGGAGTGATAGTTTGGGTCCAGGGTCCAGTGTGAGACAAAT
1838

GAG-TAG
TATTAGCCTTAGTCTATAATGCATGGATGCTCAAGTGTGAGCTCCT

GGAAACGAAATGTGCAGCTATTACAGTTGC

ATGCATTATAGACTAAG
1839

CTTAGTCTATAATGCAT
1840

Waxy starch
GTCATAGCTGCACATTTCGTTTCCAGGAGCTCACACTTGAGCATCC
1841

GBSS
ATGCATTAGAGACTTAGGCTAATAATTTGTCTCACACTGGACCCTG

Manihot esculenta

GACCCAAACTATCACTCCCAATGGTTTAA

Lys24Term
TTAAACCATTGGGAGTGATAGTTTGGGTCCAGGGTCCAGTGTGAG
1842

AAG-TAG
ACAAATTATTAGCCTAAGTCTCTAATGCATGGATGCTCAAGTGTGA

GCTCCTGGAAACGAAATGTGCAGCTATTAC

TAGAGACTTAGGCTAAT
1843

ATTAGCCTAAGTCTCTA
1844

Waxy starch
ACAACTCCTCCGTCACCGGTATAAGCATGGCAACGGTATCGATGG
1845

GBSS
CATCGTGCGTGGCGTGAAAAGGCGCGTGGAGTACAGAGACAAAA

Phaseolus vulgaris

GTGAAATCTTCGGGTCAGATGAGCCTGAACCG

Ser12Term
CGGTTCAGGCTCATCTGACCCGAAGATTTCACTTTTGTCTCTGTCC
1846

TCA-TGA
TCCACGCGCCTTTTCACGCCACGCACGATGCCATCGATACCGTTG

CCATGCTTATACCGGTGACGGAGGAGTTGT

CGTGGCGTGAAAAGGCG
1847

CGCCTTTTCACGCCACG
1848

Waxy starch
CACCGGTCTAAGCATGGCAACGGTATCGATGGCATCGTGCGTGGC
1849

GBSS
GTCAAAAGGCGCGTGAAGTACAGAGACAAAAGTGAAATCTTCGGG

Phaseolus vulgaris

TCAGATGAGCCTGAACCGTCATGAATTGAAA

Trp16Term
TTTCAATTCATGACGGTTCAGGCTCATCTGACCCGAAGATTTCACT
1850

TGG-TGA
TTTGTCTCTGTACTTCACGCGCCTTTTGACGCCACGCACGATGCCA

TCGATACCGTTGGCATGCTTATACCGGTG

GGCGCGTGAAGTACAGA
1851

TCTGTACTTCACGCGCC
1852

Waxy starch
ATAAGCATGGCAACGGTCTCGATGGCATCGTGCGTGGCGTCAAAA
1853

GBSS
GGCGCGTGGAGTACATAGACAAAAGTGAAATCTTCGGGTCAGATG

Phaseolus vulgaris

AGCCTGAACCGTCATGAATTGAAATACGATG

Glu19Term
CATCGTATTTCAATTCATGACGGTTCAGGCTCATCTGACCCGAAGA
1854

GAG-TAG
TTTCACTTTTGTCTATGTACTCCACGCGCCTTTTGACGCCACGCAC

GATGCCATCGATACCGTTGCCATGCTTAT

GGAGTACATAGACAAAA
1855

TTTTGTCTATGTACTCC
1856

Waxy starch
ATGGCAACGGTATCGATGGCATCGTGCGTGGGGTCAAAAGGCGC
1857

GBSS
GTGGAGTACAGAGACATAAGTGAAATCTTCGGGTCAGATGAGCCT

Phaseolus vulgaris

GAACCGTCATGAATTGAAATACGATGGGTTGA

Lys21Term
TCAACCCATCGTATTTCAATTCATGACGGTTCAGGCTCATCTGACC
1858

AAA-TAA
CGAAGATTTCACTTATGTCTCTGTACTCCACGCGCCTTTTGACGCC

ACGCACGATGCCATCGATACCGTTGCCAT

CAGAGACATAAGTGAAA
1859

TTTCACTTATGTCTCTG
1860

Waxy starch
ACGGTATCGATGGCATCGTGCGTGGCGTCAAAAGGCGCGTGGAG
1861

GBSS
TACAGAGACAAAAGTGTAATCTTCGGGTCAGATGAGCCTGAACCG

Phaseolus vulgaris

TCATGAATTGAAATACGATGGGTTGAGATCTC

Lys23Term
GAGATCTCAACCCATCGTATTTCAATTCATGACGGTTCAGGCTCAT
1862

AAA-TAA
CTGACCCGAAGATTACACTTTTGTCTCTGTACTCCACGCGCCTTTT

GACGCCACGCACGATGCCATCGATACCGT

CAAAAGTGTAATCTTCG
1863

CGAAGATTACACTTTTG
1864

Waxy starch
GCGCCTAGCTCGAAAAGGTCGTCATTGAGAGGCTGCACCAATGG
1865

GBSS
GTTCCATTCCTAATTAGTGTTCTTATCAAACAAACAGTGTTGGTTCA

Triticum aestivum

CTGAAACTGTCGCCTCACATCCAATTCCAG

Tyr7Term
CTGGAATTGGATGTGAGGCGACAGTTTCAGTGAACCAACACTGTT
1866

TAT-TAG
TGTTTGATAAGAACACTAATTAGGAATGGAACCCATTGGTGCAGCC

TCTCAATGACGACCTTTTCGAGCTAGGCGC

CCTAATTAGTGTTCTTA
1867

TAAGAACACTAATTAGG
1868

Waxy starch
CCTAGCTCGAAAAGGTCGTCATTGAGAGGCTGCACCAATGGGTTC
1869

GBSS
CATTCCTAATTATTGATCTTATCAAACAAACAGTGTTGGTTCACTGA

Triticum aestivum

AACTGTCGCCTCACATCCAATTCCAGCAA

Cys8Term
TTGCTGGAATTGGATGTGAGGCGACAGTTTCAGTGAACCAACACT
1870

TGT-TGA
GTTTGTTTGATAAGATCAATAATTAGGAATGGAACCCATTGGTGCA

GCCTCTCAATGACGACCTTTTCGAGCTAGG

AATTATTGATCTTATCA
1871

TGATAAGATCAATAATT
1872

Waxy starch
TCGAAAAGGTCGTCATTGAGAGGCTGCACCAATGGGTTCCATTCC
1873

GBSS
TAATTATTGTTCTTAGCAAACAAACAGTGTTGGTTCACTGAAACTGT

Triticum aestivum

CGCCTCACATCCAATTCCAGCAATCTTGT

Tyr10Term
ACAAGATTGCTGGAATTGGATGTGAGGCGACAGTTTCAGTGAACC
1874

TAT-TAG
AACACTGTTTGTTTGCTAAGAACAATAATTAGGAATGGAACCCATT

GGTGCAGCCTCTCAATGACGACCTTTTCGA

TGTTCTTAGCAAACAAA
1875

TTTGTTTGCTAAGAACA
1876

Waxy starch
CGAAAAGGTCGTCATTGAGAGGCTGCACCAATGGGTTCCATTCCT
1877

GBSS
AATTATTGTTCTTATTAAACAAACAGTGTTGGTTCACTGAAACTGTC

Triticum aestivum

GCCTCACATCCAATTCCAGCAATCTTGTA

Gln11Term
TACAAGATTGCTGGAATTGGATGTGAGGCGACAGTTTCAGTGAAC
1878

CAA-TAA
CAACACTGTTTGTTTAATAAGAACAATAATTAGGAATGGAACCCATT

GGTGCAGCCTCTCAATGACGACCTTTTCG

GTTCTTATTAAACAAAC
1879

GTTTGTTTAATAAGAAC
1880

Waxy starch
AGGCTGCACCAATGGGTTCCATTCCTAATTATTGTTCTTATCAAACA
1881

GBSS
AACAGTGTTGGTTGACTGAAACTGTCGCCTCACATCCAATTCCAGC

Triticum aestivum

AATCTTGTCACAATGAAGTTATGTTCCT

Ser17Term
AGGAACATAACTTCATTGTTACAAGATTGCTGGAATTGGATGTGAG
1882

TCA-TGA
GCGACAGTTTCAGTCAACCAACACTGTTTGTTTGATAAGAACAATA

ATTAGGAATGGAACCCATTGGTGCAGCCT

TGTTGGTTGACTGAAAC
1883

GTTTCAGTCAACCAACA
1884

Waxy starch
CAGCTCGCCACCTCCGGCACCGTCCTCGGCATCACCGACAGGTT
1885

GBSS
CCGGCGTGCAGGTTTCTAGGGCGTGAGGCCCCGGAGCCCGGCG

Triticum aestivum

GATGCGGCTCTCGGCATGAGGACCGTCGGAGCTA

Gln28Term
TAGCTCCGACGGTCCTCATGCCGAGAGCCGCATCCGCCGGGCTC
1886

CAG-TAG
CGGGGCCTCACGCCCTAGAAACCTGCACGCCGGAACCTGTCGGT

GATGCCGAGGACGGTGCCGGAGGTGGCGAGCTG

CAGGTTTCTAGGGCGTG
1887

CACGCCCTAGAAACCTG
1888

Waxy starch
GGTTTCCAGGGCGTGAGGCCCCGGAGCCCGGCGGATGCGGCTCT
1889

GBSS
CGGCATGAGGACCGTCTGAGCTAGCGCCGCCCCAACGCAAAGCC

Triticum aestivum

GGAAAGCGCACCGCGGGACCCGGCGGTGCCTCT

Gly46Term
AGAGGCACCGCCGGGTCCCGCGGTGCGCTTTCCGGCTTTGCGTT
1890

GGA-TGA
GGGGCGGCGCTAGCTCAGACGGTCCTCATGCCGAGAGCCGCATC

CGCCGGGCTCCGGGGCCTCACGCCCTGGAAACC

GGACCGTCTGAGCTAGC
1891

GCTAGCTCAGACGGTCC
1892

Waxy starch
CGGAGCCCGGCGGATGCGGCTCTCGGCATGAGGACCGTCGGAG
1893

GBSS
CTAGCGCCGCCCCAACGTAAAGCCGGAAAGCGCACCGCGGGACC

Triticum aestivum

CGGCGGTGCCTCTCCATGGTGGTGCGCGCCACCG

Gln53Term
CGGTGGCGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCG
1894

CAA-TAA
GTGCGCTTTCCGGCTTTACGTTGGGGCGGCGCTAGCTCCGACGG

TCCTCATGCCGAGAGCCGCATCCGCCGGGCTCCG

CCCCAACGTAAAGCCGG
1895

CCGGCTTTACGTTGGGG
1896

Waxy starch
GCGGATGCGGCTCTCGGCATGAGGACCGTCGGAGCTAGCGCCGC
1897

GBSS
CCCAACGCAAAGCCGGTAAGCGCACCGCGGGACCCGGCGGTGC

Triticum aestivum

CTCTCCATGGTGGTGCGCGCCACCGGCAGCGGCG

Lys56Term
CGCCGCTGCCGGTGGCGCGCACCACCATGGAGAGGCACCGCCG
1898

AAA-TAA
GGTCCCGCGGTGCGCTTACCGGCTTTGCGTTGGGGCGGCGCTAG

CTCCGACGGTCCTCATGCCGAGAGCCGCATCCGC

AAAGCCGGTAAGCGCAC
1899

GTGCGCTTACCGGCTTT
1900

Waxy starch
CTCTCCATGGTGGTGCGCGCCACCGGCAGCGGCGGCATGAACCT
1901

GBSS
CGTGTTCGTCGGCGCCTAGATGGCGCCCTGGACCAAGACCGGCG

Triticum aestivum

GCCTCGGCGACGTCCTCGGGGGCCTCCCCCCAG

Glu85Term
CTGGGGGGAGGCCCCCGAGGACGTCGCCGAGGCCGCCGGTCTT
1902

GAG-TAG
GCTCCAGGGCGCCATCTAGGCGCCGACGAACACGAGGTTCATGC

CGCCGCTGCCGGTGGCGCGCACCACCATGGAGAG

TCGGCGCCTAGATGGCG
1903

CGCCATCTAGGCGCCGA
1904

Waxy starch
GTGGTCTCTCGCTGCAGGTAGCCACACCCTGCGCGCGCGATGGC
1905

GBSS
GGCTCTGGTCACGTCGTAGCTCGCCACCTCCGGCACCGTCCTCG

Triticum aestivum

GCATCACCGACAGGTTCCGGCGTGCAGGTTTTC

Gln8Term
GAAAACCTGCACGCCGGAACCTGTCGGTGATGCCGAGGACGGTG
1906

CAG-TAG
CCGGAGGTGGCGAGCTACGACGTGACCAGAGCCGCCATCGCGC

GCGCAGGGTGTGGCTACCTGCAGCGAGAGACGAC

TCACGTCGTAGCTCGCC
1907

GGCGAGCTACGACGTGA
1908

Waxy starch
CAGCTCGCCACCTCCGGCACCGTCCTCGGCATCACCGACAGGTT
1909

GBSS
CCGGCGTGCAGGTTTTTAGGGTGTGAGGCCCCGGAGCCCGGCAG

Triticum aestivum

ATGCGCCGCTCGGCATGAGGACTACCGGAGCGA

Gln28Term
TCGCTCCGGTCGTCCTCATGCCGAGCGGCGCATCTGCCGGGCTC
1910

GAG-TAG
CGGGGCCTCACACCCTAAAAACCTGCACGCCGGAACCTGTCGGT

GATGCCGAGGACGGTGCCGGAGGTGGCGAGCTG

CAGGTTTTTAGGGTGTG
1911

CACACCCTAAAAACCTG
1912

Waxy starch
CCCCGGAGCCCGGCAGATGCGCCGCTCGGCATGAGGACTACCGG
1913

GBSS
AGCGAGCGCCGCCCCGTAGCAACAAAGCCGGAAAGCGCACCGCG

Triticum aestivum

GGACCCGGCGGTGCCTCTCCATGGTGGTGCGCG

Lys52Term
CGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCGGTGCGC
1914

AAG-TAG
TTTCCGGCTTTGTTGCTACGGGGCGGCGCTCGCTCCGGTAGTCCT

CATGCCGAGCGGCGCATCTGCCGGGCTCCGGGG

CCGCCCCGTAGCAACAA
1915

TTGTTGCTACGGGGCGG
1916

Waxy starch
CGGAGCCCGGCAGATGCGCCGCTCGGCATGAGGACTACCGGAG
1917

GBSS
CGAGCGCCGCCCCGAAGTAACAAAGCCGGAAAGCGCACCGCGG

Triticum aestivum

GACCCGGCGGTGCCTCTCCATGGTGGTGCGCGCCA

Gln53Term
TGGCGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCGGTG
1918

CAA-TAA
CGCTTTCCGGCTTTGTTACTTCGGGGCGGCGCTCGCTCCGGTAGT

CCTCATGCCGAGCGGCGCATCTGCCGGGCTCCG

CCCCGAAGTAACAAAGC
1919

GCTTTGTTACTTCGGGG
1920

Waxy starch
AGCCCGGCAGATGCGCCGCTCGGCATGAGGACTACCGGAGCGAG
1921

GBSS
CGCCGCCCCGAAGCAATAAAGCCGGAAAGCGCACCGCGGGACCC

Triticum aestivum

GGCGGTGCCTCTCCATGGTGGTGCGCGCCACGG

Gln54Term
CCGTGGCGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCG
1922

CAA-TAA
GTGCGCTTTCCGGCTTTATTGCTTCGGGGCGGCGCTCGCTCCGGT

AGTCCTCATGCCGAGCGGCGCATCTGCCGGGCT

CGAAGCAATAAAGCCGG
1923

CCGGCTTTATTGCTTCG
1924

Waxy starch
CAGCTCGCCACCTCCGGCACCGTCCTCGGCATCACCGACAGGTT
1925

GBSS
CCGGCGTGCAGGTTTCTAGGGCGTGAGGCCCCGGAACCCGGCG

Triticum durum

GATGCGGCCCTCGTCATGAGGACTATCGGAGCGA

Gln28Term
TCGCTCCGATAGTCCTCATGACGAGGGCCGCATCCGCCGGGTTC
1926

CAG-TAG
CGGGGCCTCACGCCCTAGAAACCTGCACGCCGGAACCTGTCGGT

GATGCCGAGGACGGTGCCGGAGGTGGCGAGCTG

CAGGTTTCTAGGGCGTG
1927

CACGCCCTAGAAACCTG
1928

Waxy starch
CCCCGGAACCCGGCGGATGCGGCCCTCGTCATGAGGACTATCGG
1929

GBSS
AGCGAGCGCCGCCCCGTAGCAAAGCCGGAAAGCGCACCGCGGG

Triticum durum

AGCCGGCGGTGCCTCTCCATGGTGGTGCGCGCCA

Lys52Term
TGGCGCGCACCACCATGGAGAGGCACCGCCGGCTCCCGCGGTG
1930

AAG-TAG
CGCTTTCCGGCTTTGCTACGGGGCGGCGCTCGCTCCGATAGTCCT

CATGACGAGGGCCGCATCCGCCGGGTTCCGGGG

CCGCCCCGTAGCAAAGC
1931

GCTTTGCTACGGGGCGG
1932

Waxy starch
CGGAACCCGGCGGATGCGGCCCTCGTCATGAGGACTATCGGAGC
1933

GBSS
GAGCGCCGCCCCGAAGTAAAGCCGGAAAGCGCACCGCGGGAGC

Triticum durum CGGCGGTGCCTCTCCATGGTGGTGCGCGCCACGG

Gln53Term
CCGTGGCGCGCACCACCATGGAGAGGCACCGCCGGCTCCCGCG
1934

CAA-TAA
GTGCGCTTTCCGGCTTTACTTCGGGGCGGCGCTCGCTCCGATAGT

CCTCATGACGAGGGCCGCATCCGCCGGGTTCCG

CCCCGAAGTAAAGCCGG
1935

CCGGCTTTACTTCGGGG
1936

Waxy starch
GCGGATGCGGCCCTCGTCATGAGGACTATCGGAGCGAGCGCCGC
1937

GBSS
CCCGAAGCAAAGCCGGTAAGCGCACCGCGGGAGCCGGCGGTGC

Triticum durum

CTCTCCATGGTGGTGCGCGCCACGGGCAGCGGCG

Lys56Term
CGCCGCTGCCCGTGGCGCGCACCACCATGGAGAGGCACCGCCG
1938

AAA-TAA
GCTCCCGCGGTGCGCTTACCGGCTTTGCTTCGGGGCGGGGCTCG

CTCCGATAGTCCTCATGACGAGGGCCGCATCCGC

AAAGCCGGTAAGCGCAC
1939

GTGCGCTTACCGGCTTT
1940

Waxy starch
TATCGGAGCGAGCGCCGCCCCGAAGCAAAGCCGGAAAGCGCACC
1941

GBSS
GCGGGAGCCGGCGGTGACTCTCCATGGTGGTGCGCGCCACGGG

Triticum durum

CAGCGGCGGCATGAACCTCGTGTTCGTCGGCGCC

Cys64Term
GGCGCCGACGAACACGAGGTTCATGCCGCCGCTGCCCGTGGCGC
1942

TGC-TGA
GCACCACCATGGAGAGTCACCGCCGGCTCCCGCGGTGCGCTTTC

CGGCTTTGCTTCGGGGCGGCGCTCGCTCCGATA

CGGCGGTGACTCTCCAT
1943

ATGGAGAGTCACCGCCG
1944

Waxy starch
CAGCTCGCCACCTCCGGCACCGTCCTCGGCATCACCGACAGGTT
1945

GBSS
CCGGCGTGCAGGTTTTTAGGGTGTGAGGCCCCGGAGCCCGGCAG

Triticum turgidum

ATGCGCCGCTCGGCATGAGGACTACCGGAGCGA

Gln28Term
TCGCTCCGGTAGTCCTCATGCCGAGCGGCGCATCTGCGGGGCTC
1946

CAG-TAG
CGGGGCCTCACACCCTAAAAACGTGCACGCCGGAACCTGTCGGT

GATGCCGAGGACGGTGCCGGAGGTGGCGAGCTG

CAGGTTTTTAGGGTGTG
1947

CACACCCTAAAAACCTG
1948

Waxy starch
CCCCGGAGCCCGGCAGATGCGCCGCTCGGCATGAGGACTACCGG
1949

GBSS
AGCGAGCGCCGCCCCGTAGCAACAAAGCCGGAAAGCGCACCGCG

Triticum turgidum

GGACCCGGCGGTGCCTCTCCATGGTGGTGCGCG

Lys52Term
CGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCGGTGCGC
1950

AAG-TAG
TTTCCGGCTTTGTTGCTACGGGGCGGCGCTCGCTCCGGTAGTCCT

CATGCCGAGCGGCGCATCTGCCGGGCTCCGGGG

CCGCCCCGTAGCAACAA
1951

TTGTTGCTACGGGGCGG
1952

Waxy starch
CGGAGCCCGGCAGATGCGCCGCTCGGCATGAGGACTACCGGAG
1953

GBSS
CGAGCGCCGCCCCGAAGTAACAAAGCCGGAAAGCGCACCGCGG

Triticum turgidum

GACCCGGCGGTGCCTCTCCATGGTGGTGCGCGCCA

Gln53Term
TGGCGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCGGTG
1954

CAA-TAA
CGCTTTCCGGCTTTGTTACTTCGGGGCGGCGCTCGCTCCGGTAGT

CCTCATGCCGAGCGGCGCATCTGCCGGGCTCCG

CCCCGAAGTAACAAAGC
1955

GCTTTGTTACTTCGGGG
1956

Waxy starch
AGCCCGGCAGATGCGCCGCTCGGCATGAGGACTACCGGAGCGAG
1957

GBSS
CGCCGCCCCGAAGCAATAAAGCCGGAAAGCGCACCGCGGGACCC

Triticum turgidum

GGCGGTGCCTCTCCATGGTGGTGCGCGCCACGG

Gln54Term
CCGTGGCGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCG
1958

CAA-TAA
GTGCGCTTTCCGGCTTTATTGCTTCGGGGCGGCGCTCGCTCCGGT

AGTCCTCATGCCGAGCGGCGCATCTGCCGGGCT

CGAAGCAATAAAGCCGG
1959

CCGGCTTTATTGCTTCG
1960

Waxy starch
GATGCGCCGCTCGGCATGAGGACTACCGGAGCGAGCGCCGCCCC
1961

GBSS
GAAGCAACAAAGCCGGTAAGCGCACCGCGGGACCCGGCGGTGC

Triticum turgidum

CTCTCCATGGTGGTGCGCGCCACGGGCAGCGCCG

Lys57Term
CGGCGCTGCCCGTGGCGCGCACCACCATGGAGAGGCACCGCCG
1962

AAA-TAA
GGTCCCGCGGTGCGCTTACCGGCTTTGTTGCTTCGGGGCGGCGC

TCGCTCCGGTAGTCCTCATGCCGAGCGGCGCATC

AAAGCCGGTAAGCGCAC
1963

GTGCGCTTACCGGCTTT
1964

Waxy starch
CAGCTCGCCACCTCCGCCACCGTCCTCGGCATCACCGACAGGTTC
1965

GBSS
CGCCATGCAGGTTTCTAGGGCGTGAGGCCCCGGAGCCCGGCAGA

Aegilops speltoides

TGCGCCGCTCGGCATGAGGACTGTCGGAGCGA

Gln28Term
TCGCTCCGACAGTCCTCATGCCGAGCGGCGCATCTGCCGGGCTC
1966

CAG-TAG
CGGGGCCTCACGCCCTAGAAACCTGCATGGCGGAACCTGTCGGT

GATGCCGAGGACGGTGGCGGAGGTGGCGAGCTG

CAGGTTTCTAGGGCGTG
1967

CACGCCCTAGAAACCTG
1968

Waxy starch
GGTTTCCAGGGCGTGAGGCCCCGGAGCCCGGCAGATGCGCCGCT
1969

GBSS
CGGCATGAGGACTGTCTGAGCGAGCGCCGCCCCGAAGCAACAAA

Aegilops speltoides

GCCGGAAAGCGCACCGCGGGACCCGGCGGTGCC

Gly46Term
GGCACCGCCGGGTCCCGCGGTGCGCTTTCCGGCTTTGTTGCTTC
1970

GGA-TGA
GGGGCGGCGCTCGCTCAGACAGTCCTCATGCCGAGCGGCGCATC

TGCCGGGCTCCGGGGCCTCACGCCCTGGAAACC

GGACTGTCTGAGCGAGC
1971

GCTCGCTCAGACAGTCC
1972

Waxy starch
CCCCGGAGCCCGGCAGATGCGCCGCTCGGCATGAGGACTGTCGG
1973

GBSS
AGCGAGCGCCGCCCCGTAGCAACAAAGCCGGAAAGCGCACCGCG

Aegilops speltoides

GGACCCGGCGGTGCCTCTCGATGGTGGTGCGCG

Lys52Term
CGCGCACCACCATCGAGAGGCACCGCCGGGTCCCGCGGTGCGCT
1974

AAG-TAG
TTCCGGCTTTGTTGCTACGGGGCGGCGCTCGCTCCGACAGTCCTC

ATGCCGAGCGGCGCATCTGCCGGGCTCCGGGG

CCGCCCCGTAGCAACAA
1975

TTGTTGCTACGGGGCGG
1976

Waxy starch
CGGAGCCCGGCAGATGCGCCGCTCGGCATGAGGACTGTCGGAG
1977

GBSS
CGAGCGCCGCCCCGAAGTAACAAAGCCGGAAAGCGCACCGCGG

Aegilops speltoides

GACCCGGCGGTGCCTCTCGATGGTGGTGCGCGCCA

Gln53Term
TGGCGCGCACCACCATCGAGAGGCACCGCCGGGTCCCGCGGTG
1978

CAA-TAA
CGCTTTCCGGCTTTGTTACTTCGGGGCGGCGCTCGCTCCGACAGT

CCTCATGCCGAGCGGCGCATCTGCCGGGCTCCG

CCCCGAAGTAACAAAGC
1979

GCTTTGTTACTTCGGGG
1980

Waxy starch
AGCCCGGCAGATGCGCCGCTCGGCATGAGGACTGTCGGAGCGAG
1981

GBSS
CGCCGCCCCGAAGCAATAAAGCCGGAAAGCGCACCGCGGGACCC

Aegilops speltoides

GGCGGTGCCTCTCGATGGTGGTGCGCGCCACCG

Gln54Term
CGGTGGCGCGCACCACCATCGAGAGGCACCGCCGGGTCCCGCG
1982

CAA-TAA
GTGCGCTTTCCGGCTTTATTGCTTCGGGGCGGCGCTCGCTCCGAC

AGTCCTCATGCCGAGCGGCGCATCTGCCGGGCT

CGAAGCAATAAAGCCGG
1983

CCGGCTTTATTGCTTCG
1984

Waxy starch
AGTGCAGAGATCTTCCACAGCAACAGCTAGACAACCACCATGTCG
1985

GBSS
GCTCTCACCACGTCCTAGCTCGCCACCTCGGCCACCGGCTTCGG

Oryza glaberrima

CATCGCTGACAGGTCGGCGCCGTCGTCGCTGC

Gln8Term
GCAGCGACGACGGCGCCGACCTGTCAGCGATGCCGAAGCCGGT
1986

GAG-TAG
GGCCGAGGTGGCGAGCTAGGACGTGGTGAGAGCCGACATGGTG

GTTGTCTAGCTGTTGCTGTGGAAGATCTCTGCACT

CCACGTCCTAGCTCGCC
1987

GGCGAGCTAGGACGTGG
1988

Waxy starch
TCCACAGCAACAGCTAGACAACCACCATGTCGGCTCTCACCACGT
1989

GBSS
CCCAGCTCGCCACCTAGGCCACCGGCTTCGGCATCGCTGACAGG

Oryza glaberrima

TCGGCGCCGTCGTCGCTGCTCCGCCACGGGTT

Ser12Term
AACCCGTGGCGGAGCAGCGACGACGGCGCCGACCTGTCAGCGAT
1990

TCG-TAG
GCCGAAGCCGGTGGCCTAGGTGGCGAGCTGGGACGTGGTGAGA

GCCGACATGGTGGTTGTCTAGCTGTTGCTGTGGA

CGCCACCTAGGCCACCG
1991

CGGTGGCCTAGGTGGCG
1992

Waxy starch
CGGCTCTCACCACGTCCCAGCTCGCCACCTCGGCCACCGGCTTC
1993

GBSS
GGCATCGCTGACAGGTAGGCGCCGTCGTCGCTGCTCCGCCACGG

Oryza glaberrima

GTTCCAGGGCCTCAAGCCCCGCAGCCCCGCCGG

Ser22Term
CCGGCGGGGCTGCGGGGCTTGAGGCCCTGGAACCCGTGGCGGA
1994

TCG-TAG
GCAGCGACGACGGCGCCTACCTGTCAGCGATGCCGAAGCCGGTG

GCCGAGGTGGCGAGCTGGGACGTGGTGAGAGCCG

TGACAGGTAGGCGCCGT
1995

ACGGCGCCTACCTGTCA
1996

Waxy starch
CCACGTCCCAGCTCGCCACCTCGGCCACCGGCTTCGGCATCGCT
1997

GBSS
GACAGGTCGGCGCCGTAGTCGCTGCTCCGCCACGGGTTCCAGGG

Oryza glaberrima

CCTCAAGCCCCGCAGCCCCGCCGGCGGCGACGC

Ser25Term
GCGTCGCCGCCGGCGGGGCTGCGGGGCTTGAGGCCCTGGAACC
1998

TCG-TAG
CGTGGCGGAGCAGCGACTACGGCGCCGACCTGTCAGCGATGCCG

AAGCCGGTGGCCGAGGTGGCGAGCTGGGACGTGG

GGCGCCGTAGTCGCTGC
1999

GCAGCGACTACGGCGCC
2000

Waxy starch
CGTCCCAGCTCGCCACCTCGGCCACCGGCTTCGGCATCGCTGAC
2001

GBSS
AGGTCGGCGCCGTCGTAGCTGCTCCGCCACGGGTTCCAGGGCCT

Oryza glaberrima

CAAGCCCCGCAGCCCCGCCGGCGGCGACGCGAC

Ser26Term
GTCGCGTCGCCGCCGGCGGGGCTGCGGGGCTTGAGGCCCTGGA
2002

TCG-TAG
ACCCGTGGCGGAGCAGCTACGACGGCGCCGACCTGTCAGCGATG

CCGAAGCCGGTGGCCGAGGTGGCGAGCTGGGACG

GCCGTCGTAGCTGCTCC
2003

GGAGCAGCTACGACGGC
2004

Waxy starch
TCCACAGCAAGAGCTAAACAGCCGACCGTGTGCACCACCATGTCG
2005

GBSS
GCTGTCACCACGTCCTAGCTCGCCACCTCGGCCACCGGCTTCGG

Oryza sativa

CATCGCCGACAGGTCGGCGCCGTCGTCGCTGG

Gln8Term
GCAGCGACGACGGCGCCGACCTGTCGGCGATGCCGAAGCCGGT
2006

CAG-TAG
GGCCGAGGTGGCGAGCTAGGACGTGGTGAGAGCCGACATGGTG

GTGCACACGGTCGGCTGTTTAGCTCTTGCTGTGGA

CCACGTCCTAGCTCGCC
2007

GGCGAGCTAGGACGTGG
2008

Waxy starch
CTAAACAGCCGACCGTGTGCACCACCATGTCGGCTCTCACCACGT
2009

GBSS
CCCAGCTCGCCACCTAGGCCACCGGCTTCGGCATCGCCGACAGG

Oryza sativa

TCGGCGCCGTCGTCGCTGCTTCGCCACGGGTT

Ser12Term
AACCCGTGGCGAAGCAGCGACGACGGCGCCGACCTGTCGGCGAT
2010

TCG-TAG
GCCGAAGCCGGTGGCCTAGGTGGCGAGCTGGGACGTGGTGAGA

GCCGACATGGTGGTGCACACGGTCGGCTGTTTAG

CGCCACCTAGGCCACCG
2011

CGGTGGCCTAGGTGGCG
2012

Waxy starch
CGGCTCTCACCACGTCCCAGCTCGCCACCTCGGCCACCGGCTTC
2013

GBSS
GGCATCGCCGACAGGTAGGCGCCGTCGTCGCTGCTTCGCCACGG

Oryza sativa

GTTCCAGGGCCTCAAGCCCCGTAGCCCAGCCGG

Ser22Term
CCGGCTGGGCTACGGGGCTTGAGGCCCTGGAACCCGTGGCGAA
2014

TCG-TAG
GGAGCGACGACGGCGCCTACCTGTCGGCGATGCCGAAGCCGGTG

GCCGAGGTGGCGAGCTGGGACGTGGTGAGAGCCG

CGACAGGTAGGCGCCGT
2015

ACGGCGCCTACCTGTCG
2016

Waxy starch
CCACGTCCCAGCTCGCCACCTCGGCCACCGGCTTCGGCATCGCC
2017

GBSS
GACAGGTCGGCGCCGTAGTCGCTGCTTCGCCACGGGTTCCAGGG

Oryza sativa

CCTCAAGCCCCGTAGCCCAGCCGGCGGGGACGC

Ser25Term
GCGTCCCCGCCGGCTGGGCTACGGGGCTTGAGGCCCTGGAACCC
2018

TCG-TAG
GTGGCGAAGCAGCGACTACGGCGCCGACCTGTCGGCGATGCCGA

AGCCGGTGGCCGAGGTGGCGAGCTGGGACGTGG

GGCGCCGTAGTCGCTGC
2019

GCAGCGACTACGGCGCC
2020

Waxy starch
CGTCCCAGCTCGCCACCTCGGCCACCGGCTTCGGCATCGCCGAC
2021

GBSS
AGGTCGGCGCCGTCGTAGCTGCTTCGCCACGGGTTCCAGGGCCT

Oryza sativa

CAAGCCCCGTAGCCCAGCCGGCGGGGACGCATC

Ser26Term
GATGCGTCCCCGCCGGCTGGGCTACGGGGCTTGAGGCCCTGGAA
2022

TCG-TAG
CCCGTGGCGAAGCAGCTACGACGGCGCCGACCTGTCGGCGATGC

CGAAGCCGGTGGCCGAGGTGGCGAGCTGGGACG

GCCGTCGTAGCTGCTTC
2023

GAAGCAGCTACGACGGC
2024

Waxy starch
GTCTCTCACTGCAGGTAGCCACACCCTGTGCGCGGCGCCATGGC
2025

GBSS
GGCTCTGGCCACGTCCTAGCTCGCCACCTCCGGCACCGTCCTCG

Hordeum vulgare

GCGTCACCGACAGATTCCGGCGTCCAGGTTTTC

Gln8Term
GAAAACCTGGACGCCGGAATCTGTCGGTGACGCCGAGGACGGTG
2026

GAG-TAG
CCGGAGGTGGCGAGCTAGGACGTGGCCAGAGCCGGCATGGCGC

CGCGCACAGGGTGTGGCTACCTGCAGTGAGAGAC

CCACGTCCTAGCTCGCC
2027

GGCGAGCTAGGACGTGG
2028

Waxy starch
ATGGCGGCTCTGGCCACGTCCCAGCTCGCCACGTCCGGCACCGT
2029

GBSS
CCTCGGCGTCACCGACTGATTCCGGCGTCCAGGTTTTGAGGGCCT

Hordeum vulgare

CAGGCCCCGGAACCCGGCGGATGCGGCGCTTG

Arg21Term
CAAGCGCGGCATCCGCCGGGTTCCGGGGCCTGAGGCCGTGAAAA
2030

AGA-TGA
CCTGGACGCCGGAATCAGTCGGTGACGCCGAGGACGGTGCCGG

AGGTGGCGAGCTGGGACGTGGCCAGAGCCGCCAT

TCACCGACTGATTCCGG
2031

CCGGAATCAGTCGGTGA
2032

Waxy starch
CAGCTCGCCACCTCCGGCACCGTCCTCGGCGTCACCGACAGATT
2033

GBSS
CCGGCGTCCAGGTTTTTAGGGCCTCAGGCCCCGGAACCCGGCGG

Hordeum vulgare

ATGCGGCGCTTGGTCTGAGGACTATCGGAGCAA

Gln28Term
TTGCTCCGATAGTCCTCATACCAAGCGCCGCATCCGCCGGGTTCC
2034

CAG-TAG
GGGGCCTGAGGCCCTAAAAACCTGGACGCCGGAATCTGTCGGTG

ACGCCGAGGACGGTGCCGGAGGTGGCGAGCTG

CAGGTTTTTAGGGCCTC
2035

GAGGCCCTAAAAACCTG
2036

Waxy starch
GGTTTTCAGGGCCTCAGGCCGCGGAACCCGGCGGATGCGGCGCT
2037

GBSS
TGGTATGAGGACTATCTGAGCAAGCGCCGCCCCGAAGCAAAGGC

Hordeum vulgare

GGAAAGCGGACCGCGGGAGCCGGCGGTGCCTCT

Gly46Term
AGAGGCACCGCCGGCTCCCGCGGTGCGCTTTCCGGCTTTGCTTC
2038

GGA-TGA
GGGGCGGCGCTTGCTCAGATAGTCCTCATACCAAGCGCCGCATC

CGCCGGGTTCCGGGGCCTGAGGCCCTGAAAACC

GGACTATCTGAGCAAGC
2039

GCTTGCTCAGATAGTCC
2040

Waxy starch
CCCCGGAACCCGGCGGATGCGGCGCTTGGTATGAGGACTATCGG
2041

GBSS
AGCAAGCGCCGCCCCGTAGCAAAGCCGGAAAGCGCACCGCGGG

Hordeum vulgare

AGCCGGCGGTGCCTCTCCGTGGTGGTGAGCGCCA

Lys52Term
TGGCGCTCACCACCACGGAGAGGCACCGCCGGCTCCCGCGGTGC
2042

AAG-TAG
GCTTTGCGGCTTTGCTACGGGGCGGCGCTTGCTCCGATAGTCCTC

ATACCAAGCGCCGCATCCGCCGGGTTCCGGGG

CCGCCCCGTAGCAAAGC
2043

GCTTTGCTACGGGGCGG
2044

Waxy starch
ACGTCTTTTCTCTCTCTCCTACGCAGTGGATTAATCGGCATGGCGG
2045

GBSS
CTCTGGCCACGTCGTAGCTCGTCGCAACGCGGGCCGGCCTGGGC

Zea mays

GTCCCGGACGCGTCCACGTTCCGCCGCGGCG

Gln8Term
CGCCGCGGCGGAACGTGGACGCGTCCGGGACGCCCAGGCCGGC
2046

GAG-TAG
GCGCGTTGCGACGAGCTACGACGTGGCCAGAGCCGCCATGCCGA

TTAATCCACTGCGTAGGAGAGAGAGAAAAGACGT

CCACGTCGTAGCTCGTC
2047

GACGAGCTACGACGTGG
2048

Waxy starch
GTCGCAACGCGCGCCGGCCTGGGCGTCCCGGACGCGTCCACGTT
2049

GBSS
CCGCCGCGGCGCCGCGTAGGGCCTGAGGGGGGCCCGGGCGTCG

Zea mays

GCGGGGGCGGACACGCTCAGCATGCGGACCAGCG

Gln30Term
CGCTGGTCCGCATGCTGAGCGTGTCCGCCGCCGCCGACGCCCGG
2050

CAG-TAG
GCCCCCCTCAGGCCCTACGCGGCGCCGCGGCGGAACGTGGACG

CGTCCGGGACGCCCAGGCCGGCGCGCGTTGCGAC

GCGCCGCGTAGGGCCTG
2051

CAGGCCCTACGCGGCGC
2052

Waxy starch
TCCCGGACGCGTCCACGTTCCGCCGCGGCGCCGCGCAGGGCCT
2053

GBSS
GAGGGGGGCCCGGGCGTAGGCGGCGGCGGACACGCTCAGCATG

Zea mays

CGGACCAGCGCGCGCGCGGCGCCCAGGCACCAGCA

Ser38Term
TGCTGGTGCCTGGGCGCCGCGCGCGCGCTGGTCCGCATGCTGAG
2054

TCG-TAG
CGTGTCCGCCGCCGCCTACGCCCGGGCCCCCCTCAGGCCCTGCG

CGGCGCCGCGGCGGAACGTGGACGCGTCCGGGA

CCGGGCGTAGGCGGCGG
2055

CCGCCGCCTACGCCCGG
2056

Waxy starch
GCGTCGGCGGCGGCGGACACGCTCAGCATGCGGACCAGCGCGC
2057

GBSS
GCGCGGCGCCCAGGCACTAGCAGCAGGCGCGCCGCGGGGGCAG

Zea mays

GTTCCCGTCGCTCGTCGTGTGCGCCAGCGCCGGCA

Ser57Term
TGCCGGCGCTGGCGCACACGACGAGCGACGGGAACCTGCCCCC
2058

GAG-TAG
GCGGCGCGCCTGCTGCTAGTGCCTGGGCGCCGCGCGCGCGCTG

GTCCGCATGCTGAGCGTGTCCGCCGCCGCCGACGC

CCAGGCACTAGCAGCAG
2059

CTGCTGCTAGTGCCTGG
2060

Waxy starch
TCGGCGGCGGCGGACACGCTCAGCATGCGGACCAGCGCGCGCG
2061

GBSS
CGGCGCCCAGGCACCAGTAGCAGGCGCGCCGCGGGGGCAGGTT

Zea mays

CCCGTCGCTCGTCGTGTGCGCCAGCGCCGGCATGA

Gln58Term
TCATGCCGGGGCTGGCGCACACGACGAGCGACGGGAACCTGCCC
2062

CAG-TAG
CCGCGGCGCGCCTGCTACTGGTGCCTGGGCGCCGCGCGCGCGC

TGGTCCGCATGCTGAGCGTGTCCGCCGCCGCCGA

GGCACCAGTAGCAGGCG
2063

CGCCTGCTACTGGTGCC
2064

EXAMPLE 11

Altering Fatty Acid Content of Plants

[0143] Improved means to manipulate fatty acid compositions, from biosynthetic or natural plant sources, are needed. For example, oils containing reduced saturated fatty acids are desired for dietary reasons and oils containing increased saturated fatty acids are also needed as alternatives to current sources of highly saturated oil products, such as tropical oils or chemically hydrogenated oils. It would therefore be advantageous to influence directly the production and composition of fatty acids in crop plants.

[0144] Higher plants synthesize fatty acids, primarily palmitic, stearic and oleic acids, in the plastids (i.e., chloroplasts, proplastids, or other related organelles) as part of the Fatty Acid Synthase (FAS) complex. Fatty acid synthesis is the result of the three enzymatic activities: acyl-ACP elongase, acyl-ACP desaturase and acyl-ACP thioesterases specific for each of palmitoyl-, stearoyl- and oleoyl-ACP.

[0145] A variety of enzymes have been identified that influence the relative levels of saturated vs. unsaturated fatty acids in plants. For example, the enzymes stearoyl-acyl carrier protein (stearoyl-ACP) desaturase, oleoyl desaturase and linoleate desaturase produce unsaturated fatty acids from saturated precursors. Similarly, relative enzymatic activities of the various acyl-ACP thioesterases influences the relative acyl-chain composition of the resultant fatty acids. Consequently a reduction or an increase of the activity of these enzymes can alter the properties of oils produced in a plant. In fact, specific targeting of particular enzymatic activities can results in altered levels of particular fatty acids.

[0146] The attached tables disclose exemplary oligonucleotides base sequences which can be used to generate site-specific mutations in plant genes encoding proteins involved in fatty acid biosynthesis.

24TABLE 22Oligonucleotides to produce plants with reduced palmitatePhenotype, Gene,Plant & TargetedSEQ IDAlterationAltering OligosNO:Reduced palmitateTTTGGTGGCAGTGTCTTTGAACGCTTCATCTCCTCGTCATGGTGGC2065Acyl-ACP-thioesteraseCACCTCTGCTACGTAGTCATTCTTTCCTGTACCATCTTCTTCACTTGArabidopsis thalianaATCCTAATGGAAAAGGCAATAAGATTGGSer8TermCCAATCTTATTGCCTTTTCCATTAGGATCAAGTGAAGAAGATGGTA2066TCG-TAGCAGGAAAGAATGACTACGTCGCAGAGGTGGCCACCATGACGAGGAGATGAAGCGTTCAAAGACACTGCCACCAAATGCTACGTAGTCATTCT2067AGAATGACTACGTAGCA2068Reduced palmitateGGTGGCAGTGTCTTTGAACGCTTCATCTCCTCGTCATGGTGGCCA2069Acyl-ACP-thioesteraseCCTCTGCTACGTCGTGATTCTTTCCTGTACCATCTTCTTCACTTGATArabidopsis thalianaCCTAATGGAAAAGGCAATAAGATTGGGTCSer9TermGACCCAATCTTATTGCCTTTTCCATTAGGATCAAGTGAAGAAGATG2070TCA-TGAGTACAGGAAAGAATCACGACGTAGCAGAGGTGGCCACCATGACGAGGAGATGAAGCGTTCAAAGACACTGCCACCTACGTCGTGATTCTTTC2071GAAAGAATCACGACGTA2072Reduced palmitateATCTCCTCGTCATGGTGGCCACCTCTGCTACGTCGTCATTCTTTCC2073Acyl-ACP-thioesteraseTGTACCATCTTCTTGACTTGATCCTAATGGAAAAGGCAATAAGATTArabidopsis thalianaGGGTCTACGAATCTTGCTGGACTCAATTCSer17TermGAATTGAGTCCAGCAAGATTCGTCGACCCAATCTTATTGCCTTTTC2074TCA-TGACATTAGGATCAAGTCAAGAAGATGGTCCAGGAAAGAATGACGACGTAGCAGAGGTGGCCACCATGACGAGGAGATATCTTCTTGACTTGATC2075GATCAAGTCAAGAAGAT2076Reduced palmitateGTGGCCACCTCTGCTACGTCGTCATTCTTTCCTGTACCATCTTCTT2077Acyl-AGP-thioesteraseCACTTGATCCTAATTGAAAAGGCAATAAGATTGGGTCTACGAATCTArabidopsis thalianaTGCTGGACTCAATTCTGCACCTAACTCTGGly22TermCAGAGTTAGGTGCAGAATTGAGTCCAGCAAGATTCGTCGACCCAA2078GGA-TGATCTTATTGCCTTTTCAATTAGGATCAAGTGAAGAAGATGGTCCAGGAAAGAATGACGACGTAGCAGAGGTGGCCACATCCTAATTGAAAAGGC2079GCCTTTTCAATTAGGAT2080Reduced palmitateGCTTGAATTTGTGATCTGATTGGTTAATTGTGGCCACAATGGTTGC2081Acyl-ACP-thioesteraseTACTGCCGCCACGTGATCATTCTTTCCGTTGACTTCCCCTTCTGGGGarcinia mangostanaGATGCCAAATCGGGCAATCCCGGAAAAGGSer8TermCCTTTTCCGGGATTGCCCGATTTGGCATCCCCAGAAGGGGAAGTC2082TCA-TGAAACGGAAAGAATGATCACGTGGCGGCAGTAGCAACCATTGTGGCCACAATTAACCAATCAGATCACAAATTCAAGCCGCCACGTGATCATTCT2083AGAATGATCACGTGGCG2084Reduced palmitateTGAATTTGTGATCTGATTGGTTAATTGTGGCCACAATGGTTGCTAC2085Acyl-ACP-thioesteraseTGCCGCCACGTCATGATTCTTTCCGTTGACTTCCCCTTCTGGGGATGarcinia mangostanaGCCAAATCGGGCAATCCCGGAAAAGGGTCSer9TermGACCCTTTTCCGGGATTGCCCGATTTGGCATCCCCAGAAGGGGAA2086TCA-TGAGTCAACGGAAAGAATCATGACGTGGCGGCAGTAGCAACCATTGTGGCCACAATTAACCAATCAGATCACAAATTCACACGTCATGATTCTTTC2087GAAAGAATCATGACGTG2088Reduced palmitateCTGATTGGTTAATTGTGGCCACAATGGTTGCTACTGCCGCCACGT2089Acyl-ACP-thioesteraseCATCATTCTTTCCGTAGACTTCCCCTTCTGGGGATGCCAAATCGGGGarcinia mangostanaCAATCCCGGAAAAGGGTCGGTGAGTTTTGGLeu13TermCCAAAACTCACCGACCCTTTTCCGGGATTGCCCGATTTGGCATCC2090TTG-TAGCCAGAAGGGGAAGTCTACGGAAAGAATGATGACGTGGCGGCAGTAGCAACCATTGTGGCCACAATTAACCAATCAGCTTTCCGTAGACTTCCC2091GGGAAGTCTACGGAAAG2092Reduced palmitateATGGTTGCTACTGCCGCCACGTCATCATTCTTTCCGTTGACTTCCC2093Acyl-ACP-thioesteraseCTTCTGGGGATGCCTAATCGGGCAATCCCGGAAAAGGGTCGGTGGarcinia mangostanaAGTTTTGGGTCAATGAAGTCGAAATCCGCGGLys21TermCCGCGGATTTCGACTTCATTGACCCAAAACTCACCGACCCTTTTCC2094AAA-TAAGGGATTGCCCGATTAGGCATCCCCAGAAGGGGAAGTCAACGGAAAGAATGATGACGTGGCGGCAGTCGCAACCATGGGATGCCTAATCGGGC2095GCCCGATTAGGCATCCC2096Reduced palmitateGGGATTTCAGCACGAAATTGAAGTTGTTTTTAAAAACCATGGTTGC2097Acyl-ACP-thioesteraseTACTGCTGTGACATAGGCGTTTTTCCCAGTCACTTCTTCACCTGACGossypium hirsutumTCCTCTGACTCGAAAAACAAGAAGCTCGGSer8TermCCGAGCTTCTTGTTTTTCGAGTCAGAGGAGTCAGGTGAAGAAGTG2098TCG-TAGACTGGGAAAAACGCCTATGTCACAGCAGTAGCAACCATGGTTTTTAAAAACAACTTCAATTTCGTGCTGAAATCCCTGTGACATAGGCGTTTT2099AAAACGCCTATGTCACA2100Reduced palmitateTGTTTTTAAAAACCATGGTTGCTACTGCTGTGACATCGGCGTTTTT2101Acyl-ACP-thioesteraseCCCAGTCACTTCTTGACCTGACTCCTCTGACTCGAAAAACAAGAAGGossypium hirsutumCTCGGAAGCATCAAGTCGAAGCCATCGGTSer16TermACCGATGGCTTCGACTTGATGCTTCCGAGCTTCTTGTTTTTCGAGT2102TCA-TGACAGAGGAGTCAGGTCAAGAAGTGACTGGGAAAAACGCCGATGTCACAGCAGTAGCAACCATGGTTTTTAAAAACACACTTCTTGACCTGACT2103AGTCAGGTCAAGAAGTG2104Reduced palmitateTTGCTACTGCTGTGACATCGGCGTTTTTCCCAGTCACTTCTTCACC2105Acyl-ACP-thioesteraseTGACTCCTCTGACTAGAAAAACAAGAAGCTCGGAAGCATCAAGTCGossypium hirsutumGAAGCCATCGGTTTCTTCTGGAAGTTTGCASer22TermTGCAAACTTCCAGAAGAAACCGATGGCTTCGACTTGATGCTTCCG2106TCG-TAGAGCTTCTTGTTTTTCTAGTCAGAGGAGTCAGGTGAAGAAGTGACTGGGAAAAACGCCGATGTCACAGCAGTCGCAACTCTGACTAGAAAAACA2107TGTTTTTCTAGTCAGAG2108Reduced palmitateGCTACTGCTGTGACATCGGCGTTTTTCCCAGTCACTTCTTCACCTG2109Acyl-ACP-thioesteraseACTCCTCTGACTCGTAAAACAAGAAGCTCGGAAGCATCAAGTCGAGossypium hirsutumAGCCATCGGTTTGTTCTGGAAGTTTGCAAGLys23TermCTTGCAAACTTCCAGAAGAAACCGATGGCTTCGACTTGATGCTTCC2110AAA-TAAGAGCTTCTTGTTTTACGAGTCAGAGGAGTCAGGTGAAGAAGTGACTGGGAAAAACGCCGATGTCACAGCAGTAGCCTGACTCGTAAAACAAG2111CTTGTTTTAGGAGTCAG2112Reduced palmitateCTCCCGCTCGTTGAAAGACAATGGTGGCTACCGCTGCAAGCTCTG2113Acyl-ACP-thioesteraseCATTCTTCCCCGTGTAGTCCCCGGTCACCTCCTCTAGACCAGGAACuphea hookerianaAGCCCGGAAATGGGTCATCGAGCTTCAGCCCSer14TermGGGCTGAAGCTCGATGACCCATTTCCGGGCTTTCCTGGTCTAGAG2114TCG-TAGGAGGTGACCGGGGACTACACGGGGAAGAATGCAGAGCTTGCAGCGGTAGCCACCATTGTCTTTCAACGAGCGGGAGCCCCGTGTAGTCCCCGG2115CCGGGGACTACACGGGG2116Reduced palmitateATGGTGGCTACCGCTGCAAGCTCTGCATTCTTCCCCGTGTCGTCC2117Acyl-ACP-thioesteraseCCGGTCACCTCCTCTTGACCAGGAAAGCCCGGAAATGGGTCATCGCuphea hookerianaAGCTTCAGCCCCATCAAGCCCAAATTTGTCGArg21TermCGACAAATTTGGGCTTGATGGGGCTGAAGCTCGATGACCCATTTC2118AGA-TGACGGGCTTTCCTGGTCAAGAGGAGGTGACCGGGGACGACACGGGGAAGAATGCAGAGCTTGCAGCGGTAGCCACCATCCTCCTCTTGACCAGGA2119TCCTGGTCAAGAGGAGG2120Reduced palmitateGCTACCGCTGCAAGCTCTGCATTCTTCCCCGTGTCGTCCCCGGTC2121Acyl-ACP-thioesteraseACCTCCTCTAGACCATGAAAGCCCGGAAATGGGTCATCGAGCTTCCuphea hookerianaAGCCCCATCAAGCCCAAATTTGTCGCCAATGGly23TermCATTGGCGACAAATTTGGGCTTGATGGGGCTGAAGCTCGATGACC2122GGA-TGACATTTCCGGGCTTTCATGGTCTAGAGGAGGTGACCGGGGACGACACGGGGAAGAATGCAGAGCTTGCAGCGGTAGCCTAGACCATGAAAGCCC2123GGGCTTTCATGGTCTAG2124Reduced palmitateACCGCTGCAAGCTCTGCATTCTTCCCCGTGTCGTCCCCGGTCACC2125Acyl-ACP-thioesteraseTCCTCTAGACCAGGATAGCCCGGAAATGGGTCATGGAGCTTCAGCCuphea hookerianaCCCATCAAGCCCAAATTTGTCGCCAATGGCGLys24TermCGCCATTGGCGACAAATTTGGGCTTGATGGGGCTGAAGCTCGATG2126AAG-TAGACCCATTTCCGGGCTATCCTGGTCTAGAGGAGGTGACCGGGGACGACACGGGGAAGAATGCAGAGCTTGCAGCGGTGACCAGGATAGCCCGGA2127TCCGGGCTATCCTGGTC2128Reduced palmitateGCCACCGCTGCAAGTTCTGCATTCTTCCCCCTGCCGTCCCCGGAC2129Acyl-ACP-thioesteraseACCTCCTCTAGGCCGTGAAAGCTGGGAAATGGGTCATCGAGCTTGCuphea lanceolataAGCCCCCTCAAGCCCAAATTTGTCGCCAATGGly23TermCATTGGCGACAAATTTGGGCTTGAGGGGGCTCAAGCTCGATGACC2130GGA-TGACATTTCCGAGCTTTCACGGCCTAGAGGAGGTGTCCGGGGACGGCAGGGGGAAGAATGCAGAACTTGCAGCGGTGGCCTAGGCCGTGAAAGCTC2131GAGCTTTCACGGCCTAG2132Reduced palmitateACCGCTGCAAGTTCTGCATTCTTCCCCCTGCCGTCCCCGGACACC2133Acyl-ACP-thioesteraseTCCTCTAGGCCGGGATAGCTCGGAAATGGGTCATCGAGCTTGAGCCuphea lanceolataCCCCTCAAGCCCAAATTTGTCGCCAATGCCGLys24TermCGGCATTGGCGACAAATTTGGGCTTGAGGGGGCTCAAGCTCGAT2134AAG-TAGGACCCATTTCCGAGCTATCCCGGCCTAGAGGAGGTGTCCGGGGACGGCAGGGGGAAGAATGCAGAACTTGCAGCGGTGGCCGGGATAGCTCGGA2135TCCGAGCTATCCCGGCC2136Reduced palmitateGCAAGTTCTGCATTCTTCCCCCTGCCGTCCCCGGACACCTCCTCT2137Acyl-ACP-thioesteraseAGGCCGGGAAAGCTCTGAAATGGGTCATCGAGCTTGAGCCCCCTCuphea lanceolataCAAGCCCAAATTTGTCGCCAATGCCGGGTTGAGly26TermTCAACCCGGCATTGGCGACAAATTTGGGGTTGAGGGGGCTCAAGC2138GGA-TGATCGATGACCCATTTCAGAGCTTTCCCGGCCTAGAGGAGGTGTCCGGGGACGGCAGGGGGAAGAATGCAGAACTTGCGAAAGCTCTGAAATGGG2139CCCATTTCAGAGCTTTC2140Reduced palmitateCATTCTTCCCCCTGCCGTCCCCGGACACCTCCTCTAGGCCGGGAA2141Acyl-ACP-thioesteraseAGCTCGGAAATGGGTGATCGAGCTTGAGCCCCCTCAAGCCCAAATCuphea lanceolataTTGTCGCCAATGCCGGGTTGAAGGTTAAGGCSer29TermGCCTTAACCTTCAACCCGGCATTGGCGACAAATTTGGGCTTGAGG2142TCA-TGAGGGCTCAAGCTCGATCACCCATTTCCGAGCTTTCCCGGCCTAGAGGAGGTGTCCGGGGACGGCAGGGGGAAGAATGAAATGGGTGATCGAGCT2143AGCTCGATCACCCATTT2144Reduced palmitateCGTTTAAGTGGATCGGACATTTAAGTGTTTTAATCATGGTAGCTAT2145Acyl-ACP-thioesteraseGAGTGCTACTGCGTAGCTGTTTCCGGTTTCTTCCCCAAAACCTCACHelianthus annuusTCTGGAGCCAAGACATCTGATAAGCTTGGSer9TermCCAAGCTTATCAGATGTCTTGGCTCCAGAGTGAGGTTTTGGGGAA2146TCG-TAGGAAACCGGAAACAGCTACGCAGTCGCACTCATAGCTACCATGATTAAAACACTTAAATGTCCGATCCACTTAAACGTACTGCGTAGCTGTTTC2147GAAACAGCTACGCAGTA2148Reduced palmitateAGTGTTTTAATCATGGTCGCTATGAGTGCTACTGCGTCGCTGTTTC2149Acyl-ACP-thioesteraseCGGTTTCTTCCCCATAACCTCACTCTGGAGCCAAGACATCTGATAAHelianthus annuusGCTTGGAGGTGAACCAGGTAGTGTTGCTGLys17TermCAGCAACACTACCTGGTTCACCTCCAAGCTTATCAGATGTCTTGGC2150AAA-TAATCCAGAGTGAGGTTATGGGGAAGAAACCGGAAACAGCGACGCAGTAGCACTCATAGCTACCATGATTAAAACACTCTTCCCCATAACCTCAC2151GTGAGGTTATGGGGAAG2152Reduced palmitateATGGTAGCTATGAGTGCTACTGCGTCGCTGTTTCCGGTTTCTTCCC2153Acyl-ACP-thioesteraseCAAAACCTCACTCTTGAGCCAAGACATCTGATAAGCTTGGAGGTGHelianthus annuusAACCAGGTAGTGTTGCTGTGCGCGGAATCAGly21TermTGATTCCGCGCACAGCAACACTACCTGGTTCACCTCCAAGCTTATC2154GGA-TGAAGATGTCTTGGCTCAAGAGTGAGGTTTTGGGGAAGAAACCGGAAACAGCGACGCAGTAGCACTCATAGCTACCATCTCACTCTTGAGCCAAG2155CTTGGCTCAAGAGTGAG2156Reduced palmitateGCTATGAGTGCTACTGCGTCGCTGTTTCCGGTTTCTTCCCCAAAAC2157Acyl-ACP-thioesteraseCTCACTCTGGAGCCTAGACATCTGATAAGCTTGGAGGTGAACCAGHelianthus annuusGTAGTGTTGCTGTGCGCGGAATCAAGACAALys23TermTTGTCTTGATTCCGCGCACAGCAACACTACCTGGTTCACCTCCAAG2158AAG-TAGCTTATCAGATGTCTAGGCTCCAGAGTGAGGTTTTGGGGAAGAAACCGGAAACAGCGACGCAGTCGCACTCATAGCCTGGAGCCTAGACATCT2159AGATGTCTAGGCTCCAG2160Reduced palmitateATGGTGGCTGCTGCAGCAAGTTCTGCATGCTTCCCTGTTCCATCC2161Acyl-ACP-thioesteraseCCAGGAGCCTCCCCTTAACCTGGGAAGTTAGGCAACTGGTCATCGCuphea palustrisAGTTTGAGCCCTTCCTTGAAGCCCAAGTCAALys21TermTTGACTTGGGCTTCAAGGAAGGGCTCAAACTCGATGACCAGTTGC2162AAA-TAACTAACTTCCCAGGTTAAGGGGAGGCTCCTGGGGATGGAACAGGGAAGCATGCAGAACTTGCTGCAGCAGCCACCATCCTCCCCTTAACCTGGG2163CCCAGGTTAAGGGGAGG2164Reduced palmitateGCTGCAGCAAGTTCTGCATGCTTCCCTGTTCCATCCCCAGGAGCC2165Acyl-ACP-thioesteraseTCCCCTAAACCTGGGTAGTTAGGCAACTGGTCATCGAGTTTGAGCCuphea palustrisCCTTCCTTGAAGCCCAAGTCAATCCCCAATGLys24TermCATTGGGGATTGACTTGGGCTTCAAGGAAGGGCTCAAACTCGATG2166AAG-TAGACCAGTTGCCTAACTACCCAGGTTTAGGGGAGGCTCCTGGGGATGGAACAGGGAAGCATGCAGAACTTGCTGCAGCAACCTGGGTAGTTAGGC2167GCCTAACTACCCAGGTT2168Reduced palmitateTGCATGCTTCCCTGTTCCATCCCCAGGAGCCTCCCCTAAACCTGG2169Acyl-ACP-thioesteraseGAAGTTAGGCAACTGATCATCGAGTTTGAGCCCTTCCTTGAAGCCCuphea palustrisCAAGTCAATCCCCAATGGCGGATTTCAGGTTTrp28TermAACCTGAAATCCGCCATTGGGGATTGACTTGGGCTTCAAGGAAGG2170TGG-TGAGCTCAAACTCGATGATCAGTTGCCTAACTTCCCAGGTTTAGGGGAGGCTCCTGGGGATGGAACAGGGAAGCATGCAGGCAACTGATCATCGAG2171CTCGATGATCAGTTGCC2172Reduced palmitateCATGCTTCCCTGTTCCATCCCCAGGAGCCTCCCCTAAACCTGGGA2173Acyl-ACP-thioesteraseAGTTAGGCAACTGGTGATCGAGTTTGAGCCCTTCCTTGAAGCCCACuphea palustrisAGTCAATCCCCAATGGCGGATTTCAGGTTAASer29TermTTAACCTGAAATCCGCCATTGGGGATTGACTTGGGCTTCAAGGAA2174TCA-TGAGGGCTCAAACTCGATCACCAGTTGCCTAACTTCCCAGGTTTAGGGGAGGCTCCTGGGGATGGAACAGGGAAGCATGCAACTGGTGATCGAGTT2175AACTCGATCACCAGTTG2176Reduced paimitateATGGTGGCTGCCGCAGCAAGTTCTGCATTCTTCTCCGTTCCAACC2175Acyl-ACP-thioesteraseCCGGGAATCTCCCCTTAACCCGGGAAGTTCGGTAATGGTGGCTTTCuphea hookerianaCAGGTTAAGGCAAACGCCAATGCCCATCCTA Lys21TermTAGGATGGGCATTGGCGTTTGCCTTAACCTGAAAGCCACCATTAC2178AAA-TAACGAACTTCCCGGGTTAAGGGGAGATTCCCGGGGTTGGAACGGAGAAGAATGCAGAACTTGCTGCGGCAGCCACCATTCTCCCCTTAACCCGGG2179CCCGGGTTAAGGGGAGA2180Reduced palmitateGCCGCAGCAAGTTCTGCATTCTTCTCCGTTCCAACCCCGGGAATC2181Acyl-ACP-thioesteraseTCCCCTAAACCCGGGTAGTTCGGTAATGGTGGCTTTCAGGTTAAGCuphea hookerianaGCAAACGCCAATGCCCATCCTAGTCTAAAGTLys24TermACTTTAGACTAGGATGGGCATTGGCGTTTGCCTTAACCTGAAAGC2182AAG-TAGCACCATTACCGAACTACCCGGGTTTAGGGGAGATTCCCGGGGTTGGAACGGAGAAGAATGCAGAACTTGCTGCGGCAACCCGGGTAGTTCGGT2183ACCGAACTACCCGGGTT2184Reduced palmitateTTCTCCGTTCCAACCCCGGGAATCTCCCCTAAACCCGGGAAGTTC2185Acyl-ACP-thioesteraseGGTAATGGTGGCTTTTAGGTTAAGGCAAACGCCAATGCCCATCCTCuphea hookerianaAGTCTAAAGTCTGGCAGCCTCGAGACTGAAGGln31TermCTTCAGTCTCGAGGCTGCCAGACTTTAGACTAGGATGGGCATTGG2186CAG-TAGCGTTTGCCTTAACCTAAAAGCCACCATTACCGAACTTCCCGGGTTTAGGGGAGATTCCCGGGGTTGGAACGGAGAAGTGGCTTTTAGGTTAAG2187CTTAACCTAAAAGCCAC2188Reduced palmitateGTTCCAACCCCGGGAATCTCCCCTAAACCCGGGAAGTTCGGTAAT2189Acyl-ACP-thioesteraseGGTGGCTTTCAGGTTTAGGCAAACGCCAATGCCCATCCTAGTCTACuphea hookerianaAAGTCTGGCAGCCTCGAGACTGAAGATGACALys33TermTGTCATCTTCAGTCTCGAGGCTGCCAGACTTTAGACTAGGATGGG2190AAG-TAGCATTGGCGTTTGCCTAAACCTGAAAGCCACCATTACCGAACTTCCCGGGTTTAGGGGAGATTCCCGGGGTTGGAACTTCAGGTTTAGGCAAAC2191GTTTGCCTAAACCTGAA2192Reduced palmitateATGTTGAAGCTCTCGTGTAATGCGACTGATAAGTTACAGACCCTCT2193Acyl-ACP-thioesteraseTCTCGCATTCTCATTAACCGGATCCGGCACACCGGAGAACCGTCTBrassica rapaCCTCCGTGTCGTGCTCTCATCTGAGGAAACGln21TermGTTTCCTCAGATGAGAGCACGACACGGAGGAGACGGTTCTCCGGT2194CAA-TAAGTGCCGGATCCGGTTAATGAGAATGCGAGAAGAGGGTCTGTAACTTATCAGTCGCATTACACGAGAGCTTCAACATATTCTCATTAACCGGAT2195ATCCGGTTAATGAGAAT2196Reduced palmitateGCGACTGATAAGTTACAGACCCTCTTCTCGCATTCTCATCAACCGG2197Acyl-ACP-thioesteraseATCCGGCACACCGGTGAACCGTCTCCTCCGTGTCGTGCTCTCATCBrassica rapaTGAGGAAACCGGTTCTCGATCCTTTGCGAGArg28TermCTCGCAAAGGATCGAGAACCGGTTTCCTCAGATGAGAGCACGACA2198AGA-TGACGGAGGAGACGGTTCACCGGTGTGCCGGATCCGGTTGATGAGAATGCGAGAAGAGGGTCTGTAACTTATCAGTCGCCACACCGGTGAACCGTC2199GACGGTTCACCGGTGTG2200Reduced palmitateCCCTCTTCTCGCATTCTCATCAACCGGATCCGGCACACCGGAGAA2201Acyl-ACP-thioesteraseCCGTCTCCTCCGTGTAGTGCTCTCATCTGAGGAAACCGGTTCTCGBrassica rapaATCCTTTGCGAGCGATCGTATCTGCTGATCASer24TermTGATCAGCAGATACGATCGCTCGCAAAGGATCGAGAACCGGTTTC2202TCG-TAGCTCAGATGAGAGCACTACACGGAGGAGACGGTTCTCCGGTGTGCCGGATCCGGTTGATGAGAATGCGAGAAGAGGGCTCCGTGTAGTGCTCTC2203GAGAGCACTACACGGAG2204Reduced palmitateCTTCTCGCATTCTCATCAACCGGATCCGGCACACCGGAGAACCGT2205Acyl-ACP-thioesteraseCTCCTCCGTGTCGTGATCTCATCTGAGGAAACCGGTTCTCGATCCBrassica rapaTTTGCGAGCGATCGTATCTGCTGATCAAGGACys25TermTCCTTGATCAGCAGATACGATCGCTCGCAAAGGATCGAGAACCGG2206TGC-TGATTTCCTCAGATGAGATCACGACACGGAGGAGACGGTTCTCCGGTGTGCCGGATCCGGTTGATGAGAATGCGAGAAGGTGTCGTGATCTCATCT2207AGATGAGATCACGACAC2208Reduced palmitateATTCTTCTTCTATAAACCAAAACCTCAGGAACCATAAAAAAAAAAGG2209Acyl-ACP-thioesteraseGCATCAAAAATGTAGAAGCTTTCGTGTAATGTGACTAACAACTTACBrassica napusACACCTTCTCCTTCTTCTCCGATTCCTCLeu2TermGAGGAATCGGAGAAGAAGGAGAAGGTGTGTAAGTTGTTAGTCACA2210TTG-TAGTTACACGAAAGCTTCTACATTTTTGATGCCCTTTTTTTTTTATGGTTCCTGAGGTTTTGGTTTATAGAAGAAGAATAAAAATGTAGAAGCTTT2211AAAGCTTCTACATTTTT2212Reduced palmitateTCTTCTTCTATAAACCAAAACCTCAGGAACCATAAAAAAAAAAGGG2213Acyl-ACP-thioesteraseCATCAAAAATGTTGTAGCTTTCGTGTAATGTGACTAACAACTTACACBrassica napusACCTTCTCCTTCTTCTCCGATTCCTCCCLys3TermGGGAGGAATCGGAGAAGAAGGAGAAGGTGTGTAAGTTGTTAGTCA2214AAG-TAGCATTACACGAAAGCTACAACATTTTTGATGCCCTTTTTTTTTTATGGTTCCTGAGGTTTTGGTTTATAGAAGAAGAAAATGTTGTAGCTTTCG2215CGAAAGCTACAACATTT2216Reduced palmitateCTATAAACCAAAACCTCAGGAACCATAAAAAAAAAAGGGCATCAAA2217Acyl-ACP-thioesteraseAATGTTGAAGCTTTAGTGTAATGTGACTAACAACTTACACACCTTCTBrassica napusCCTTCTTCTCCGATTCCTCCCTTTTCATSer5TermATGAAAAGGGAGGAATCGGAGAAGAAGGAGAAGGTGTGTAAGTT2218TCG-TAGGTTAGTCACATTACACTAAAGCTTCAACATTTTTGATGCCCTTTTTTTTTTATGGTTCCTGAGGTTTTGGTTTATAGGAAGCTTTAGTGTAATG2219CATTACACTAAAGCTTC2220Reduced palmitateAAACCAAAACCTCAGGAACCATAAAAAAAAAAGGGCATCAAAAATG2221Acyl-ACP-thioesteraseTTGAAGCTTTCGTGAAATGTGACTAACAACTTACACACCTTCTCCTTBrassica napusCTTCTCCGATTCCTCCCTTTTCATCCCGCys6TermCGGGATGAAAAGGGAGGAATCGGAGAAGAAGGAGAAGGTGTGTA2222TGT-TGAAGTTGTTAGTCACATTTCACGAAAGCTTCAACATTTTTGATGCCCTTTTTTTTTTATGGTTCCTGAGGTTTTGGTTTCTTTCGTGAAATGTGAC2223GTCACATTTCACGAAAG2224

[0147]

25

TABLE 23

Oligonucleotides to produce plants with increased stearate

Phenotype, Gene,

Plant & Targeted

SEQ ID

Alteration
Altering Oligos
NO:

Increased stearate
GGGAGAGCTCTAGCTCTGTAGAAAAGAAGGATTCATTCATCATATC
2225

stearoyl-ACP
CAGAAATGGCTCTATAGTTTAACCCTTTGGTGGCATCTCAGCCTTA

desaturase
CAAATTCCCTTCCTCGACTCGTCCGCCAA

Arabidopsis thaliana

TTGGCGGACGAGTCGAGGAAGGGAATTTGTAAGGCTGAGATGCC
2226

Lys4 Term
ACCAAAGGGTTAAACTATAGAGCCATTTCTGGATATGATGAATGAA

AAG-TAG
TCCTTCTTTTCTACAGAGCTAGAGCTCTCCC

TGGCTCTATAGTTTAAC
2227

GTTAAACTATAGAGCCA
2228

Increased stearate
CTCTGTAGAAAAGAAGGATTCATTCATCATATCCAGAAATGGCTCT
2229

stearoyl-ACP
AAAGTTTAACCCTTAGGTGGCATCTCAGCCTTACAAATTCCCTTCC

desaturase
TCGACTCGTCCGCCAACTCCTCTTTCAG

Arabidopsis thaliana

CTGAAAGAAGGAGTTGGCGGACGAGTCGAGGAAGGGAATTTGTA
2230

Leu8 Term
AGGCTGAGATGCCACCTAAGGGTTAAACTTTAGAGCCATTTCTGG

TTG-TAG
ATATGATGAATGAATCCTTCTTTTCTACAGAG

TAACCCTTAGGTGGCAT
2231

ATGCCACCTAAGGGTTA
2232

Increased stearate
AGAAGGATTCATTCATCATATCCAGAAATGGCTCTAAAGTTTAACC
2233

stearoyl-ACP
CTTTGGTGGCATCTTAGCCTTACAAATTCCCTTCCTCGACTCGTCC

desaturase
GCCAACTCCTTCTTTCAGATCTCCCAAGT

Arabidopsis thaliana

ACTTGGGAGATCTGAAAGAAGGAGTTGGCGGACGAGTCGAGGAA
2234

Gln12 Term
GGGAATTTGTAAGGCTAAGATGCCACCAAAGGGTTAAACTTTAGA

CAG-TAG
GCCATTTCTGGATATGATGAATGAATCCTTCT

TGGCATCTTAGCCTTAC
2235

GTAAGGCTAAGATGCCA
2236

Increased stearate
TCATTCATCATATCCAGAAATGGCTCTAAAGTTTAACCCTTTGGTG
2237

stearoyl-ACP
GCATCTCAGCCTTAGAAATTCCCTTCCTCGACTCGTCCGCCAACTC

desaturase
CTTCTTTCAGATCTCCCAAGTTCCTCTGC

Arabidopsis thaliana

GCAGAGGAACTTGGGAGATCTGAAAGAAGGAGTTGGCGGACGAG
2238

Phe14 Term
TCGAGGAAGGGAATTTCTAAGGCTGAGATGCCACCAAAGGGTTAA

TAC-TAG
ACTTTAGAGCCATTTCTGGATATGATGAATGA

CAGCCTTAGAAATTCCC
2239

GGGAATTTCTAAGGCTG
2240

Increased stearate
GAGAGCTCGCTCGTGTCTGAAAGAACATCAAACCTCGTATCAAAAA
2241

stearoyl-ACP
AAAGAAAATGGCATAGAAGCTTAACCCTTTGGCATCTCAGCCTTAC

desaturase
AAACTCCCTTCCTCGGCTCGTCCGCCAAT

Brassica napus

ATTGGCGGACGAGCCGAGGAAGGGAGTTTGTAAGGCTGAGATGC
2242

Leu3 Term
CAAAGGGTTAAGCTTCTATGCCATTTTCTTTTTTTTGATACGAGGTT

TTG-TAG
TGATGTTCTTTCAGACACGAGCGAGCTCTC

AATGGCATAGAAGCTTA
2243

TAAGCTTCTATGCCATT
2244

Increased stearate
GAGCTCGCTCGTGTCTGAAAGAACATCAAACCTCGTATCAAAAAAA
2245

stearoyl-ACP
AGAAAATGGCATTGTAGCTTAACCCTTTGGCATCTCAGCCTTACAA

desaturase
ACTCCCTTCCTCGGCTCGTCCGCCAATCT

Brassica napus

AGATTGGCGGACGAGCCGAGGAAGGGAGTTTGTAAGGCTGAGAT
2246

Lys4 Term
GCCAAAGGGTTAAGCTACAATGCCATTTTCTTTTTTTTGATACGAG

AAG-TAG
GTTTGATGTTCTTTCAGACACGAGCGAGCTC

TGGCATTGTAGCTTAAC
2247

GTTAAGCTACAATGCCA
2248

Increased stearate
TCTGAAAGAACATCAAACCTCGTATCAAAAAAAAGAAAATGGCATT
2249

stearoyl-ACP
GAAGCTTAACCCTTAGGCATCTCAGCCTTACAAACTCCCTTCCTCG

desaturase
GCTCGTCCGCCAATCTCTACTCTCAGATC

Brassica napus

GATCTGAGAGTAGAGATTGGCGGACGAGCCGAGGAAGGGAGTTT
2250

Leu8 Term
GTAAGGCTGAGATGCCTAAGGGTTAAGCTTCAATGCCATTTTCTTT

TTG-TAG
TTTTTGATACGAGGTTTGATGTTCTTTCAGA

TAACCCTTAGGCATCTC
2251

GAGATGCCTAAGGGTTA
2252

Increased stearate
AACATCAAACCTCGTATCAAAAAAAAGAAAATGGCATTGAAGCTTA
2253

stearoyl-ACP
ACCCTTTGGCATCTTAGCCTTACAAACTCCCTTCCTCGGCTCGTCC

desaturase
GCCAATCTCTACTCTCAGATCTCCCAAGT

Brassica napus

ACTTGGGAGATCTGAGAGTAGAGATTGGCGGACGAGCCGAGGAA
2254

Gln11 Term
GGGAGTTTGTAAGGCTAAGATGCCAAAGGGTTAAGCTTCAATGCC

CAG-TAG
ATTTTCTTTTTTTTGATACGAGGTTTGATGTT

TGGCATCTTAGCCTTAC
2255

GTAAGGCTAAGATGCCA
2256

Increased stearate
AACCAAAAGAAAAGGTAAGAAAAAAAACAATGGCTCTCAAGCTCA
2257

stearoyl-ACP
ATCCTTTCCTTTCTTAAACCCAAAAGTTACCTTCTTTCGCTCTTCCA

desaturase
CCAATGGCCAGTACCAGATCTCCTAAGT

Ricinus communis

ACTTAGGAGATCTGGTACTGGCCATTGGTGGAAGAGCGAAAGAAG
2258

Gln27 Term
GTAACTTTTGGGTTTAAGAAAGGATTGAGCTTGAGAGCCAT

CAA-TAA
TGTTTTTTTTCTTACCTTTTTCTTTTGGTT

TCCTTTCTTAAACCCAA
2259

TTGGGTTTAAGAAAGGA
2260

Increased stearate
AAGAAAAAGGTAAGAAAAAAAACAATGGCTCTCAAGCTCAATCCTT
2261

stearoyl-ACP
TCCTTTCTCAAACCTAAAAGTTACCTTCTTTCGCTCTTCCACCAATG

desaturase
GCCAGTACCAGATCTCCTAAGTTCTACA

Ricinus communis

TGTAGAACTTAGGAGATCTGGTACTGGCCATTGGTGGAAGAGCGA
2262

Gln29 Term
AAGAAGGTAACTTTTAGGTTTGAGAAAGGAAAGGATTGAGCTTGA

CAA-TAA
GAGCCATTGTTTTTTTTCTTACCTTTTTCTT

CTCAAACCTAAAAGTTA
2263

TAACTTTTAGGTTTGAG
2264

Increased stearate
AAAAAGGTAAGAAAAAAAACAATGGCTCTCAAGCTCAATCCTTTCC
2265

stearoyl-ACP
TTTCTCAAACCCAATAGTTACCTTCTTTCGCTCTTCCACCAATGGCC

desaturase
AGTACCAGATCTCCTAAGTTCTACATGG

Ricinus communis

CCATGTAGAACTTAGGAGATCTGGTACTGGCCATTGGTGGAAGAG
2266

Lys30 TermCGAAAGAAGGTAACTATTGGGTTTGAGAAAGGAAAGGATTGAGCT

AAG-TAG
TGAGAGCCATTGTTTTTTTTCTTACCTTTTT

AAACCCAATAGTTACCT
2267

AGGTAACTATTGGGTTT
2268

Increased stearate
TCTCAAACCCAAAAGTTACCTTCTTTCGCTCTTCCACCAATGGCCA
2269

stearoyl-ACP
GTACCAGATCTCCTTAGTTCTACATGGCCTCTACCCTCAAGTCTGG

desaturase
TTCTAAGGAAGTTGAGAATCTCAAGAAGC

Ricinus communis

GCTTCTTGAGATTCTCAACTTCCTTAGAACCAGACTTGAGGGTAGA
2270

Lys46 Term
GGCCATGTAGAACTAAGGAGATCTGGTACTGGCCATTGGTGGAG

AAG-TAG
AGCGAAAGAAGGTAACTTTTGGGTTTGAGA

GATCTCCTTAGTTCTAC
2271

GTAGAACTAAGGAGATC
2272

Increased stearate
TCTTCTGATTCATTTAATCTTTACTCATCAATGGCTCTGAGACTGAA
2273

stearoyl-ACP
CCCTATCCCCACCTAAACCTTCTCCCTCCCCCAAATGGCCAGTCTC

desaturase
AGATCTCCCAGGTTCCGCATGGCCTCTA

Glycine max

TAGAGGCCATGCGGAACCTGGGAGATCTGAGACTGGCCATTTGG
2274

Gln11 Term
GGGAGGGAGAAGGTTTAGGTGGGGATAGGGTTCAGTCTCAGAGC

CAA-TAA
CATTGATGAGTAAAGATTAAATGAATCAGAAGA

TCCCCACCTAAACCTTC
2275

GAAGGTTTAGGTGGGGA
2276

Increased stearate
CTTTACTCATCAATGGCTCTGAGACTGAACCCTATCCCCACCCAAA
2277

stearoyl-ACP
CCTTCTCCCTCCCCTAAATGGCCAGTCTCAGATCTCCCAGGTTCC

desaturase
GCATGGCCTCTACCCTCCGCTCCGGTTCCA

Glycine max

TGGAACCGGAGCGGAGGGTAGAGGCCATGCGGAACCTGGGAGAT
2278

Gln17 Term
CTGAGACTGGCCATTTAGGGGAGGGAGAAGGTTTGGGTGGGGAT

CAA-TAA
AGGGTTCAGTCTCAGAGCCATTGATGAGTAAAG

CCCTCCCCTAAATGGCC
2279

GGCCATTTAGGGGAGGG
2280

Increased stearate
GCTCTGAGACTGAACCCTATCCCCACCCAAACCTTCTCCCTCCCC
2281

stearoyl-ACP
CAAATGGCCAGTCTCTGATCTCCCAGGTTCCGCATGGCCTCTACC

desaturase
CTCCGCTCCGGTTCCAAAGAGGTTGAAAATA

Glycine max

TATTTTCAACCTCTTTGGAACCGGAGCGGAGGGTAGAGGCCATGC
2282

Arg22 Term
GGAACCTGGGAGATCAGAGACTGGCCATTTGGGGGAGGGAGAAG

AGA-TGA
GTTTGGGTGGGGATAGGGTTCAGTCTCAGAGC

CCAGTCTCTGATCTCCC
2283

GGGAGATCAGAGACTGG
2284

Increased stearate
CAAATGGCCAGTCTCAGATCTCCCAGGTTCCGCATGGCCTCTACC
2285

stearoyl-ACP
CTCCGCTCCGGTTCCTAAGAGGTTGAAAATATTAAGAAGCCATTCA

desaturase
CTCCTCCCAGAGAAGTGCATGTTCAAGTAA

Glycine max

TTACTTGAACATGCACTTCTCTGGGAGGAGTGAATGGCTTCTTAAT
2286

Lys37 Term
ATTTTCAACCTCTTAGGAACCGGAGCGGAGGGTAGAGGCCATGCG

AAA-TAA
GAACCTGGGAGATCTGAGACTGGCCATTTG

CCGGTTCCTAAGAGGTT
2287

AACCTCTTAGGAACCGG
2288

Increased stearate
CAACAAGCACACACAAGAACAACATCAACAATGGCGATTCGCATC
2289

stearoyl-ACP
AATACGGCGACGTTTTAATCAGACCTGTACCGTTCATTCGCGTTTC

desaturase
CTCAACCGAAACCTCTCAGATCTCCCAAAT

Helianthus annuus

ATTTGGGAGATCTGAGAGGTTTCGGTTGAGGAAACGCGAATGAAC
2290

Gln11 Term
GGTACAGGTCTGATTAAAACGTCGCCGTATTGATGCGAATCGCCA

CAA-TAA
TTGTTGATGTTGTTCTTGTGTGTGCTTGTTG

CGACGTTTTAATCAGAC
2291

GTCTGATTAAAACGTCG
2292

Increased stearate
AAGCACACACAAGAAGCAACATCAACAATGGCGATTCGCATCAATAC
2293

stearoyl-ACP
GGCGACGTTTCAATGAGACCTGTACCGTTCATTCGCGTTTCCTCAA

desaturase
CCGAAACCTCTCAGATCTCCCAAATTCGC

Helianthus annuus

GCGAATTTGGGAGATCTGAGAGGTTTCGGTTGAGGAAACGCGAAT
2294

Ser12 Term
GAACGGTACAGGTCTCATTGAAACGTCGCCGTATTGATGCGAATC

TCA-TGA
GCCATTGTTGATGTTGTTCTTGTGTGTGCTT

GTTTCAATGAGACCTGT
2295

ACAGGTCTCATTGAAAC
2296

Increased stearate
AAGAACAACATCAACAATGGCGATTCGCATCAATACGGCGACGTTT
2297

stearoyl-ACP
CAATCAGACCTGTAGCGTTCATTCGCGTTTCCTCAACCGAAACCTC

desaturase
TCAGATCTCCCAAATTCGCCATGGCTTCC

Helianthus annuus

GGAAGCCATGGCGAATTTGGGAGATCTGAGAGGTTTCGGTTGAGG
2298

Tyr15 Term
AAACGCGAATGAACGCTACAGGTCTGATTGAAACGTCGCCGTATT

TAC-TAG
GATGCGAATCGCCATTGTTGATGTTGTTCTT

GACCTGTAGCGTTCATT
2299

AATGAACGCTACAGGTC
2300

Increased stearate
CAACATCAACAATGGCGATTCGCATCAATACGGCGACGTTTCAATC
2301

stearoyl-ACP
AGACCTGTACCGTTGATTCGCGTTTCCTCAACCGAAACCTCTCAGA

desaturase
TCTCCCAAATTCGCCATGGCTTCCACCAT

Helianthus annuus

ATGGTGGAAGCCATGGCGAATTTGGGAGATCTGAGAGGTTTCGGT
2302

Ser17 Term
TGAGGAAACGCGAATCAACGGTACAGGTCTGATTGAAACGTCGCC

TCA-TGA
GTATTGATGCGAATCGCCATTGTTGATGTTG

GTACCGTTGATTCGCGT
2303

ACGCGAATCAACGGTAC
2304

Increased stearate
ACACACAACACACACTCAATCACACACACATCATCATCTTCTTCATC
2305

stearoyl-ACP
AACGATGGCGCTTTGAATGAGTCCGGTGACGCTTCAACGGGAGAT

desaturase
ATATCCTTCATACACTTTTCATCAATCGA

Helianthus annuus

TCGATTGATGAAAAGTGTATGAAGGATATATCTCCCGTTGAAGCGT
2306

Arg4 Term
CACCGGACTCATTCAAAGCGCCATCGTTGATGAAGAAGATGATGA

CGA-TGA
TGTGTGTGTGATTGAGTGTGTGTTGTGTGT

TGGCGCTTTGAATGAGT
2307

ACTCATTCAAAGCGCCA
2308

Increased stearate
ACACACACATCATCATCTTCTTCATCAACGATGGCGCTTCGAATGA
2309

stearoyl-ACP
GTCCGGTGACGCTTTAACGGGAGATATATCCTTCATACACTTTTCA

desaturase
TCAATCGAAAAATCTCAGATCTCCTAAAT

Helianthus annuus

ATTTAGGAGATCTGAGATTTTTCGATTGATGAAAAGTGTATGAAGG
2310

Gln11 Term
ATATATCTCCCGTTAAAGCGTCACCGGACTCATTCGAAGCGCCATC

CAA-TAA
GTTGATGAAGAAGATGATGATGTGTGTGT

TGACGCTTTAACGGGAG
2311

CTCCCGTTAAAGCGTCA
2312

Increased stearate
ACATCATCATCTTCTTCATCAACGATGGCGCTTCGAATGAGTCCGG
2313

stearoyl-ACP
TGACGCTTCAACGGTAGATATATCCTTCATACACTTTTCATCAATCG

desaturase
AAAAATCTCAGATCTCCTAAATTCGCGA

Helianthus annuus

TCGCGAATTTAGGAGATCTGAGATTTTTCGATTGATGAAAAGTGTA
2314

Glu13 Term
TGAAGGATATATCTACCGTTGAAGCGTCACCGGACTCATTCGAAG

GAG-TAG
CGCCATCGTTGATGAAGAAGATGATGATGT

TTCAACGGTAGATATAT
2315

ATATATCTACCGTTGAA
2316

Increased stearate
ATCTTCTTCATCAACGATGGCGCTTCGAATGAGTCCGGTGACGCTT
2317

stearoyl-ACP
CAACGGGAGATATAGCCTTCATACACTTTTCATCAATCGAAAAATC

desaturase
TCAGATCTCCTAAATTCGCGATGGCTTCC

Helianthus annuus

GGAAGCCATCGCGAATTTAGGAGATCTGAGATTTTTCGATTGATGA
2318

Tyr15 Term
AAAGTGTATGAAGGCTATATCTCCCGTTGAAGCGTCACCGGACTC

TAT-TAG
ATTCGAAGCGCCATCGTTGATGAAGAAGAT

GAGATATAGCCTTCATA
2319

TATGAAGGCTATATCTC
2320

Increased stearate
AACTCAGCCAGCTTGCCCCCAAACAACAGCGCAGAAAAACCTTCA
2321

stearoyl-ACP
ACAACAATGGCTCTCTAGCTCAACCCAGTCACCACCTTCCCTTCAA

desaturase
CACGCTCCCTCAACAACTTCTCCTCCAGAT

Linum usitatissimum

ATCTGGAGGAGAAGTTGTTGAGGGAGCGTGTTGAAGGGAAGGTG
2322

Lys4 Term
GTGACTGGGTTGAGCTAGAGAGCCATTGTTGTTGAAGGTTTTTCT

AAG-TAG
GCGCTGTTGTTTGGGGGCAAGCTGGCTGAGTT

TGGCTCTCTAGCTCAAC
2323

GTTGAGCTAGAGAGCCA
2324

Increased stearate
GCGCAGAAAAACCTTCAACAACAATGGCTCTCAAGCTCAACCCAG
2325

stearoyl-ACP
TCACCACCTTCCCTTGAACACGCTCCCTCAACAACTTCTCCTCCAG

desaturase
ATCTCCTCGCACCTTTCTCATGGCTGCTTC

Linum usitatissimum

GAAGCAGCCATGAGAAAGGTGCGAGGAGATCTGGAGGAGAAGTT
2326

Ser13 Term
GTTGAGGGAGCGTGTTCAAGGGAAGGTGGTGACTGGGTTGAGCT

TCA-TGA
TGAGAGCCATTGTTGTTGAAGGTTTTTCTGCGC

CTTCCCTTGAACACGCT
2327

AGCGTGTTCAAGGGAAG
2328

Increased stearate
CTCAAGCTCAACCCAGTCACCACCTTCCCTTCAACACGCTCCCTCA
2329

stearoyl-ACP
ACAACTTCTCCTCCTGATCTCCTCGCACCTTTCTCATGGCTGCTTC

desaturase
CACTTTCAATTCCACCTCCACCAAGTAAG

Linum usitatissimum

CTTACTTGGTGGAGGTGGAATTGAAAGTGGAAGCAGCCATGAGAA
2330

Arg23 Term
AGGTGCGAGGAGATCAGGAGGAGAAGTTGTTGAGGGAGCGTGTT

AGA-TGA
GAAGGGAAGGTGGTGACTGGGTTGAGCTTGAG

TCTCCTCCTGATCTCCT
2331

AGGAGATCAGGAGGAGA
2332

Increased stearate
TCCTCCAGATCTCCTCGCACCTTTCTCATGGCTGCTTCCACTTTCA
2333

stearoyl-ACP
ATTCCACCTCCACCTAGTAAGCATCTCCTCCTCCTCGGAATCTCCG

desaturase
CCGATTTCTTTTAAGCGATTGATCGTAGA

Linum usitatissimum

TCTACGATCAATCGCTTAAAAGAAATCGGCGGAGATTCCGAGGAG
2334

Lys411 Term
GAGGAGATGCTTACTAGGTGGAGGTGGAATTGAAAGTGGAAGCA

AAG-TAG
GCCATGAGAAAGGTGCGAGGAGATCTGGAGGA

CCTCCACCTAGTAAGCA
2335

TGCTTACTAGGTGGAGG
2336

Increased stearate
ATGGCACTGAAACTTTGCTTTCCACCCCACAAGATGCCTTCCTTCC
2337

stearoyl-ACP
CCGATGCTCGTATCTGATCTCACAGGGTTTTCATGGCTTCAACTAT

desaturase
TCATTCTCCTTCTATGGAGGTCGGAAAAG

Olea europaeap

CTTTCCGACCTCCATAGAAGGAGAATGAATAGTTGAAGCCATGAA
2338

Arg21 Term
AACCCTGTGAGATCAGATACGAGCATCGGGGAAGGAAGGCATCTT

AGA-TGA
GTGGGGTGGAAAGCAAAGTTTCAGTGCCAT

CTCGTATCTGATCTCAC
2339

GTGAGATCAGATACGAG
2340

Increased stearate
CCCACAAGATGCCTTCCTTCCCCGATGCTCGTATCAGATCTCACAG
2341

stearoyl-ACP
GGTTTTCATGGCTTGAACTATTCATTCTCCTTCTATGGAGGTCGGA

desaturase
AAAGTTAAAAAGCCTTTCACGCCTCCACG

Olea europaeap

CGTGGAGGCGTGAAAGGCTTTTTAACTTTTCCGACCTCCATAGAA
2342

Ser29 Term
GGAGAATGAATAGTTCAAGCCATGAAAACCCTGTGAGATCTGATAC

TCA-TGA
GAGCATCGGGGAAGGAAGGCATCTTGTGGG

CATGGCTTGAACTATTC
2343

GAATAGTTCAAGCCATG
2344

Increased stearate
GATGCTCGTATCAGATCTCACAGGGTTTTCATGGCTTCAACTATTC
2345

stearoyl-ACP
ATTCTCCTTCTATGTAGGTCGGAAAAGTTAAAAAGCCTTTCACGCC

desaturase
TCCACGAGAGGTACATGTTCAAGTAACCC

Olea europaeap

GGGTTACTTGAACATGTACCTCTCGTGGAGGCGTGAAAGGCTTTT
2346

Glu37 Term
TAACTTTTCCGACCTACATGAAGGAGAATGAATAGTTGAAGCCAT

GAG-TAG
GAAAACCCTGTGAGATCTGATACGAGCATC

CTTCTATGTAGGTCGGA
2347

TCCGACCTACATAGAAG
2348

Increased stearate
CGTATCAGATCTCACAGGGTTTTCATGGCTTCAACTATTCATTCTC
2349

stearoyl-ACP
CTTCTATGGAGGTCTGAAAAGTTAAAAAGCCTTTCACGCCTCCACG

desaturase
AGAGGTACATGTTCAAGTAACCCATTCCT

Olea europaeap

AGGAATGGGTTACTTGAACATGTACCTCTCGTGGAGGCGTGAAAG
2350

Gly39 Term
GCTTTTTAACTTTTCAGACCTCCATAGAAGGAGAATGAATAGTTGA

GGA-TGA
AGCCATGAAAACCCTGTGAGATCTGATACG

TGGAGGTCTGAAAAGTT
2351

AACTTTTCAGACCTCCA
2352

Increased stearate
TTCTCGTTTTTGTCGTCCCCTCTGCTCTCTCTCTCTATCAGGCACG
2353

stearoyl-ACP
GAGAAATGGCACTGTAACTCAGTCCAGTCATGTTTCAATCTCAGAA

desaturase
GCTTCCATTTCTTGCCTCCTATCCGCCTT

Persea americana

AAGGCGGATAGGAGGCAAGAAATGGAAGCTTCTGAGATTGAAACA
2354

Lys4 Term
TGACTGGACTGAGTTACAGTGCCATTTCTCCGTGCCTGATAGAGA

AAA-TAA
GAGAGAGCAGAGGGGACGACAAAAACGAGAA

TGGCACTGTAACTCAGT
2355

ACTGAGTTACAGTGCCA
2356

Increased stearate
CTGCTCTCTCTCTCTATCAGGCACGGAGAAATGGCACTGAAACTCA
2357

stearoyl-ACP
GTCCAGTCATGTTTTAATCTCAGAAGCTTCCATTTCTTGCCTCCTAT

desaturase
CCGCCTTCCAATCTCAGATCTCCGAGGG

Persea americana

CCCTCGGAGATCTGAGATTGGAAGGCGGATAGGAGGCAAGAAAT
2358

Gln11 Term
GGAAGCTTCTGAGATTAAAACATGACTGGACTGAGTTTCAGTGCC

CAA-TAA
ATTTCTCCGTGCCTGATAGAGAGAGAGAGCAG

TCATGTTTTAATCTCAG
2359

CTGAGATTAAAACATGA
2360

Increased stearate
TCTCTCTCTATCAGGCACGGAGAAATGGCACTGAAACTCAGTCCA
2361

stearoyl-ACP
GTCATGTTTCAATCTTAGAAGCTTCCATTTCTTGCCTCCTATCCGCC

desaturase
TTCCAATCTCAGATCTCCGAGGGTTTTCA

Persea americana

TGAAAACCCTCGGAGATCTGAGATTGGAAGGCGGATAGGAGGCAA
2362

Gln13 Term
GAAATGGAAGCTTCTAAGATTGAAACATGACTGGACTGAGTTTCAG

CAG-TAG
TGCCATTTCTCCGTGCCTGATAGAGAGAGA

TTCAATCTTAGAAGCTT
2363

AAGCTTCTAAGATTGAA
2364

Increased stearate
CTCTCTATCAGGCACGGAGAAATGGCACTGAAACTCAGTCCAGTC
2365

stearoyl-ACP
ATGTTTCAATCTCAGTAGCTTCCATTTCTTGCCTCCTATCCGCCTTC

desaturase
CAATCTCAGATCTCCGAGGGTTTTCATGG

Persea americana

CCATGAAAACCCTCGGAGATCTGAGATTGGAAGGCGGATAGGAG
2366

Lys14 Term
GCAAGAAATGGAAGCTACTGAGATTGAAACATGACTGGACTGAGT

AAG-TAG
TTCAGTGCCATTTCTCCGTGCCTGATAGAGAG

AATCTCAGTAGCTTCCA
2367

TGGAAGCTACTGAGATT
2368

Increased stearate
CCCCGAGATCTCGCTGCCGCTGCTCATGGCGTTCGCGGCGTCCC
2369

stearoyl-ACP
ACACCGCATCGCCGTAGTCCTGCGGCGGCGTGGCGCAGAGGAG

desaturase
GAGCAATGGGATGTCGAAGATGGTGGCCATGGCC

Oryza sativa

GGCCATGGCCACCATCTTCGACATCCCATTGCTCCTCCTCTGCGC
2370

Tyr12 Term
CACGCCGCCGCAGGACTACGGCGATGCGGTGTGGGACGCCGCG

TAC-TAG
AACGCCATGAGCAGCGGCAGCGAGATCTCGGGG

TCGCCGTAGTCCTGCGG
2371

CCGCAGGACTACGGCGA
2372

Increased stearate
CTGCTCATGGCGTTCGCGGCGTCCCACACCGCATCGCCGTACTCC
2373

stearoyl-ACP
TGCGGCGGCGTGGCGTAGAGGAGGAGCAATGGGATGTCGAAGAT

desaturase
GGTGGCCATGGCCTCCACCATCAACAGGGTCA

Oryza sativa

TGACCCTGTTGATGGTGGAGGCCATGGCCACCATCTTCGACATCC
2374

Gln19 Term
CATTGCTCCTCCTCTACGCCACGCCGCCGCAGGAGTACGGCGAT

CAG-TAG
GCGGTGTGGGACGCCGCGAACGCCATGAGCAG

GCGTGGCGTAGAGGAGG
2375

CCTCCTCTACGCCACGC
2376

Increased stearate
CCCACACCGCATCGCCGTACTCCTGCGGCGGCGTGGCGCAGAGG
2377

stearoyl-ACP
AGGAGCAATGGGATGTAGAAGATGGTGGCCATGGCCTCCACCAT

desaturase
CAACAGGGTCAAGACTGCTAAGAAGCCCTACAC

Oryza sativa

GTGTAGGGCTTCTTAGCAGTCTTGACCCTGTTGATGGTGGAGGCC
2378

Ser26 Term
ATGGCCACCATCTTCTACATCCCATTGCTCCTCCTCTGCGCCACGC

TCG-TAG
CGCCGCAGGAGTACGGCGATGCGGTGTGGG

TGGGATGTAGAAGATGG
2379

CCATCTTCTACATCCCA
2380

Increased stearate
CACACCGCATCGCCGTACTCCTGCGGCGGCGTGGCGCAGAGGAG
2381

stearoyl-ACP
GAGCAATGGGATGTCGTAGATGGTGGCCATGGCCTCCACCATCAA

desaturase
CAGGGTCAAGACTGCTAAGAAGCCCTACACTC

Oryza sativa

GAGTGTAGGGCTTCTTAGCAGTCTTGACCCTGTTGATGGTGGAGG
2382

Lys27 Term
CCATGGCCACCATCTACGACATCCCATTGCTCCTCCTCTGCGCCA

AAG-TAG
CGCCGCCGCAGGAGTACGGCGATGCGGTGTG

GGATGTCGTAGATGGTG
2383

CACCATCTACGACATCC
2384

Increased stearate
TTCTCTCTCTAGGTTGAGCGGTTACCAACAGAAGCACTTAGGAGA
2385

stearoyl-ACP
GAGAAGCAATGGCGTAGAAGCTTCACCACACGGCCTTCAATCCTT

desaturase
CCATGGCGGTTACCTCTTCGGGACTTCCTCG

Simmondsia chinensis

CGAGGAAGTCCCGAAGAGGTAACCGCCATGGAAGGATTGAAGGC
2386

Leu3 Term
CGTGTGGTGAAGCTTCTACGCCATTGCTTCTCTCTCCTAAGTGCTT

TTG-TAG
CTGTTGGTAACCGCTCAACCTAGAGAGAGAA

AATGGCGTAGAAGCTTC
2387

GAAGCTTCTACGCCATT
2388

Increased stearate
CTCTCTCTAGGTTGAGCGGTTACCAACAGAAGCACTTAGGAGAGA
2389

stearoyl-ACP
GAGCAATGGCGTTGTAGCTTCACCACACGGCCTTCAATCCTTCC

desaturase
ATGGCGGTTACCTCTTCGGGACTTCCTCGAT

Simmondsia chinensis

ATCGAGGAAGTCCCGAAGAGGTAACCGCCATGGAAGGATTGAAG
2390

Lys4 Term
GCCGTGTGGTGAAGCTACAACGCCATTGCTTCTCTCTCCTAAGTG

AAG-TAG
CTTCTGTTGGTAACCGCTCAACCTAGAGAGAG

TGGCGTTGTAGCTTCAC
2391

GTGAAGCTACAACGCCA
2392

Increased stearate
AAGCAATGGCGTTGAAGCTTCACCACACGGCCTTCAATCCTTCCAT
2393

stearoyl-ACP
GGCGGTTACCTCTTAGGGACTTCCTCGATCGTATCACCTCAGATCT

desaturase
CACCGCGTTTTCATGGCTTCTTCTACAAT

Simmondsia chinensis

ATTGTAGAAGAAGCCATGAAAACGCGGTGAGATCTGAGGTGATAC
2394

Ser19 Term
GATCGAGGAAGTCCCTAAGAGGTAACCGCCATGGAAGGATTGAAG

TCG-TAG
GCCGTGTGGTGAAGCTTCAACGCCATTGCTT

TACCTCTTAGGGACTTC
2395

GAAGTCCCTAAGAGGTA
2396

Increased stearate
GCAATGGCGTTGAAGCTTCACCACACGGCCTTCAATCCTTCCATG
2397

stearoyl-ACP
GCGGTTACCTCTTCGTGACTTCCTCGATCGTATCACCTCAGATCTC

desaturase
ACCGCGTTTTCATGGCTTCTTCTACAATTG

Simmondsia chinensis

CAATTGTAGAAGAAGCCATGAAAACGCGGTGAGATCTGAGGTGAT
2398

Gly20 Term
ACGATCGAGGAAGTCACGAAGAGGTAACCGCCATGGAAGGATTG

GGA-TGA
AAGGCCGTGTGGTGAAGCTTCAACGCCATTGC

CCTCTTCGTGACTTCCT
2399

AGGAAGTCACGAAGAGG
2400

Increased stearate
TGGCTCTGAATCTCAACCCCGTTTCCACACCATTTCAGTGTCGTCG
2401

stearoyl-ACP
ATTGCCGTCTTTCTGACCTCGTCAAACGCCTTCTCGCAGATCTCCC

desaturase
AAATTCTTCATGGCTTCCACTCTCAGCAG

Spinacia oleracea

CTGCTGAGAGTGGAAGCCATGAAGAATTTGGGAGATCTGCGAGAA
2402

Ser21 Term
GGCGTTTGACGAGGTCAGAAAGACGGCAATCGACGACACTGAAAT

TCA-TGA
GGTGTGGAAACGGGGTTGAGATTCAGAGCCA

GTCTTTCTGACCTCGTC
2403

GACGAGGTCAGAAAGAC
2404

Increased stearate
AATCTCAACCCCGTTTCCACACCATTTCAGTGTCGTCGATTGCCGT
2405

stearoyl-ACP
CTTTCTCACCTCGTTAAACGCCTTCTCGCAGATCTCCCAAATTCTT

desaturase
CATGGCTTCCACTCTCAGCAGCTCTTCTC

Spinacia oleracea

GAGAAGAGCTGCTGAGAGTGGAAGCCATGAAGAATTTGGGAGATC
2406

Gln24 Term
TGCGAGAAGGCGTTTAACGAGGTGAGAAAGACGGCAATCGACGA

CAA-TAA
CACTGAAATGGTGTGGAAACGGGGTTGAGATT

CACCTCGTTAAACGCCT
2407

AGGCGTTTAACGAGGTG
2408

Increased stearate
TCCACACCATTTCAGTGTCGTCGATTGCCGTCTTTCTCACCTCGTC
2409

stearoyl-ACP
AAACGCCTTCTCGCTGATCTCCCAAATTCTTCATGGCTTCCACTCT

desaturase
CAGCAGCTCTTCTCCTAAGGAAGCGGAAA

Spinacia oleracea

TTTCCGCTTCCTTAGGAGAAGAGCTGCTGAGAGTGGAAGCCATGA
2410

Arg29 Term
AGAATTTGGGAGATCAGCGAGAAGGCGTTTGACGAGGTGAGAAA

AGA-TGA
GACGGCAATCGACGACACTGAAATGGTGTGGA

CTTCTCGCTGATCTCCC
2411

GGGAGATCAGCGAGAAG
2412

Increased stearate
TTTCAGTGTCGTCGATTGCCGTCTTTCTCACCTCGTCAAACGCCTT
2413

stearoyl-ACP
CTCGCAGATCTCCCTAATTCTTCATGGCTTCCACTCTCAGCAGCTC

desaturase
TTCTCCTAAGGAAGCGGAAAGCCTGAAGA

Spinacia oleracea

TCTTCAGGCTTTCCGCTTCCTTAGGAGAAGAGCTGCTGAGAGTGG
2414

Lys32 Term
AAGCCATGAAGAATTAGGGAGATCTGCGAGAAGGCGTTTGACGAG

AAA-TAA
GTGAGAAAGACGGCAATCGACGACACTGAAA

GATCTCCCTAATTCTTC
2415

GAAGAATTAGGGAGATC
2416

Increased stearate
AAATAGTCGAGGTGAAAAACAGAGCATCAACAATGGCACTGAATAT
2417

stearoyl-ACP
CAATGGGGTGTCGTGAAAATCTCACAAAATGTTACCATTTCCTTGT

desaturase
TCTTCAGCCAGATCTGAGCGAGTTTTCAT

Solanum tuberosum

ATGAAAACTCGCTCAGATCTGGCTGAAGAACAAGGAAATGGTAAC
2418

Leu10 Term
ATTTTGTGAGATTTTCACGACACCCCATTGATATTCAGTGCCATTGT

TTA-TGA
TGATGCTCTGTTTTTCACCTCGACTATTT

GGTGTCGTGAAAATCTC
2419

GAGATTTTCACGACACC
2420

Increased stearate
ATAGTCGAGGTGAAAACAGAGCATCAACAATGGCACTGAATATCA
2421

stearoyl-ACP
ATGGGGTGTCGTTATAATCTCACAAAATGTTACCATTTCCTTGTTCT

desaturase
TCAGCCAGATCTGAGCGAGTTTTCATGG

Solanum tuberosum

CCATGAAAACTCGCTCAGATCTGGCTGAAGAACAAGGAAATGGTA
2422

Lys11 Term
ACATTTTGTGAGATTATAACGACACCCCATTGATATTCAGTGCCATT

AAA-TAA
GTTGATGCTCTGTTTTTCACCTCGACTAT

TGTCGTTATAATCTCAC
2423

GTGAGATTATAACGACA
2424

Increased stearate
GTGAAAAACAGAGCATCAACAATGGCACTGAATATCAATGGGGTG
2425

stearoyl-ACP
TCGTTAAAATCTCACTAAATGTTACCATTTCCTTGTTCTTCAGCCAG

desaturase
ATCTGAGCGAGTTTTCATGGCTTCAACCA

Solanum tuberosum

TGGTTGAAGCCATGAAAACTCGCTCAGATCTGGCTGAAGAACAAG
2426

Lys14 Term
GAAATGGTAACATTTAGTGAGATTTTAACGACACCCCATTGATATT

AAA-TAA
CAGTGCCATTGTTGATGCTCTGTTTTTCAC

AATCTCACTAAATGTTA
2427

TAACATTTAGTGAGATT
2428

Increased stearate
ACAGAGCATCAACAATGGCACTGAATATCAATGGGGTGTCGTTAAA
2429

stearoyl-ACP
ATCTCACAAAATGTGACCATTTCCTTGTTCTTCAGCCAGATCTGAG

desaturase
CGAGTTTTCATGGCTTCAACCATTCATCG

Solanum tuberosum

CGATGAATGGTTGAAGCCATGAAAACTCGCTCAGATCTGGCTGAA
2430

Leu16 Term
GAACAAGGAAATGGTCACATTTTGTGAGATTTTAACGACACCCCAT

TTA-TGA
TGATATTCAGTGCCATTGTTGATGCTCTGT

CAAAATGTGACCATTTC
2431

GAAATGGTCACATTTTG
2432

Increased stearate
TGGCTCTGAGGCTGAACCCTAACCCTTCACAGAAGCTCTTTCTCTC
2433

stearoyl-ACP
TCCTTCTTCATCATGATCTTCTTCTTCTTCATCGTTCTCGCTTCCTC

desaturase
AAATGGCTAGCCTCAGATCTCCAAGGTT

Arachis hypogaea

AACCTTGGAGATCTGAGGCTAGCCATTTGAGGAAGCGAGAACGAT
2434

Ser21 Term
GAAGAAGAAGAAGATCATGATGAAGAAGGAGAGAGAAAGAGCTTC

TCA-TGA
TGTGAAGGGTTAGGGTTCAGCCTCAGAGCCA

TTCATCATGATCTTCTT
2435

AAGAAGATCATGATGAA
2436

Increased stearate
ACCCTAACCCTTCACAGAAGCTCTTTCTCTCTCCTTCTTCATCATCA
2437

stearoyl-ACP
TCTTCTTCTTCTTGATCGTTCTCGCTTCCTCAAATGGCTAGCCTCA

desaturase
GTCTCCAAGGTTCCGCATGGCCTCCAC

Arachis hypogaea

GTGGAGGCCATGCGGAACCTTGGAGATCTGAGGCTAGCCATTTGA
2438

Ser26 Term
GGAAGCGAGAACGATCAAGAAGAAGAAGATGATGATGAAGAAGGA

TCA-TGA
GAGAGAAAGAGCTTCTGTGAAGGGTTAGGGT

TTCTTCTTGATCGTTCT
2439

AGAACGATCAAGAAGAA
2440

Increased stearate
CTAACCCTTCACAGAAGCTCTTTCTCTCTCCTTCTTCATCATCATCT
2441

stearoyl-ACP
TCTTCTTCTTCATAGTTCTCGCTTCCTCAAATGGCTAGCCTCAGAT

desaturase
CTCCAAGGTTCCGCATGGCCTCCACCCT

Arachis hypogaea

AGGGTGGAGGCCATGCGGAACCTTGGAGATCTGAGGCTAGCCAT
2442

Ser27 Term
TTGAGGAAGCGAGAACTATGAAGAAGAAGAAGATGATGATGAAGA

TCG-TAG
AGGAGAGAGAAAGAGCTTCTGTGAAGGGTTAG

TTCTTCATAGTTCTCGC
2443

GCGAGAACTATGAAGAA
2444

Increased stearate
CTTCACAGAAGCTCTTTCTCTCTCCTTCTTCATCATCATCTTCTTCT
2445

stearoyl-ACP
TCTTCATCGTTCTAGCTTCCTCAAATGGCTAGCCTCAGATCTCCAA

desaturase
GGTTCCGCATGGCCTCCACCCTCCGCAC

Arachis hypogaea

GTGCGGAGGGTGGAGGCCATGCGGAACCTTGGAGATCTGAGGCT
2446

Ser29 Term
AGCCATTTGAGGAAGCTAGAACGATGAAGAAGAAGAAGATGATGA

TCG-TAG
TGAAGAAGGAGAGAGAAAGAGCTTCTGTGAAG

ATCGTTCTAGCTTCCTC
2447

GAGGAAGCTAGAACGAT
2448

Increased stearate
AAAGTTAAAAGCCGTCCAAAACCCAAACCAGGAAAGGCAAACGAA
2449

stearoyl-ACP
AAGAAAAAATGGCTTAGAATTTTAATGCCATCGCCTCGAAATCTCA

desaturase
GAAGCTCCCTTGCTTTGCTCTTCCACCAAA

Gossypium hirsutum

TTTGGTGGAAGAGCAAAGCAAGGGAGCTTCTGAGATTTCGAGGCG
2450

Leu3 Term
ATGGCATTAAAATTCTAAGCCATTTTTTCTTTTCGTTTGCCTTTCCT

TTG-TAG
GGTTTGGGTTTTGGACGGCTTTTAACTTT

AATGGCTTAGAATTTTA
2451

TAAAATTCTAAGCCATT
2452

Increased stearate
CCCAAACCAGGAAAGGCAAACGAAAAGAAAAAATGGCTTTGAATTT
2453

stearoyl-ACP
TAATGCCATCGCCTAGAAATCTCAGAAGCTCCCTTGCTTTGCTCTT

desaturase
CCACCAAAGGCCACCCTTAGATCTCCCAA

Gossypium hirsutum

TTGGGAGATCTAAGGGTGGCCTTTGGTGGAAGAGCAAAGCAAGG
2454

Ser1-Term
GAGCTTCTGAGATTTCTAGGCGATGGCATTAAAATTCAAAGCCATT

TCG-TAG
TTTTCTTTTCGTTTGCCTTTCCTGGTTTGGG

CATCGCCTAGAAATCTC
2455

GAGATTTCTAGGCGATG
2456

Increased stearate
CAAACCAGGAAAGGCAAACGAAAAGAAAAAATGGCTTTGAATTTTA
2457

stearoyl-ACP
ATGCCATCGCCTCGTAATCTCAGAAGCTCCCTTGCTTTGCTCTTCC

desaturase
ACCAAAGGCCACCCTTAGATCTCCCAAGT

Gossypium hirsutum

ACTTGGGAGATCTAAGGGTGGCCTTTGGTGGAAGAGCAAAGCAAG
2458

Lys11 Term
GGAGCTTCTGAGATTACGAGGCGATGGCATTAAAATTCAAAGCCAA

AAA-TAA
TTTTTTCTTTTCGTTTGCCTTTCCTGGTTTG

TCGCCTCGTAATCTCAG
2459

CTGAGATTACGAGGCGA
2460

Increased stearate
AGGAAAGGCAAACGAAAAGAAAAAATGGCTTTGAATTTTAATGCCA
2461

stearoyl-ACP
TCGCCTCGAAATCTTAGAAGCTCCCTTGCTTTGCTCTTCCACCAAA

desaturase
GGCCACCCTTAGATCTCCCAAGTTTTCCA

Gossypium hirsutum

TGGAAAACTTGGGAGATCTAAGGGTGGCCTTTGGTGGAAGAGCAA
2462

Gln13 Term
AGCAAGGGAGCTTCTAAGATTTCGAGGCGATGGCATTAAAATTCA

CAG-TAG
AAGCCATTTTTTCTTTTCGTTTGCCTTTCCT

CGAAATCTTAGAAGCTC
2463

GAGCTTCTAAGATTTCG
2464

[0148]

26

TABLE 24

Oligonucleotides to produce plants with reduced linolenic acid

Phenotype, Gene,

Plant & Targeted

SEQ ID

Alteration
Altering Oligos
NO:

Reducing linolenic acid
AATAGAACGACAGAGACTTTTTCCTCTTTTCTTCTTGGGAAGAGGC
2465

omega-3 fatty acid
TCCAATGGCGAGCTAGGTTTTATCAGAATGTGGTTTTAGACCTCTC

desaturase
CCCAGATTCTACCCTAAACACACAACCTC

Arabidopsis thaliana

GAGGTTGTGTGTTTAGGGTAGAATCTGGGGAGAGGTCTAAAACCA
2466

Ser4 Term
CATTCTGATAAAACCTAGCTCGCCATTGGAGCCTCTTCCCAAGAAG

TCG-TAG
AAAAGAGGAAAAAGTCTCTGTCGTTCTATT

GGCGAGCTTGGTTTTAT
2467

ATAAAACCAAGCTCGCC
2468

Reducing linolenic acid
ACGACAGAGACTTTTTCCTCTTTTCTTCTTGGGAAGAGGCTCCAAT
2469

omega-3 fatty acid
GGCGAGCTCGGTTTGATCAGAATGTGGTTTTAGACCTCTCCCCAG

desaturase
ATTCTACCCTAAACACACAACCTCTTTTGC

Arabidopsis thaliana

GCAAAAGAGGTTGTGTGTTTAGGGTAGAATCTGGGGAGAGGTCTA
2470

Leu6 Term
AAACCACATTCTGATCAAACCGAGCTCGCCATTGGAGCCTCTTCCC

TTA-TGA
AAGAAGAAAAGAGGAAAAAGTCTCTGTCGT

CTCGGTTTGATCAGAAT
2471

ATTCTGATCAAACCGAG
2472

Reducing linolenic acid
ACAGAGACTTTTTCCTCTTTTCTTCTTGGGAAGAGGCTCCAATGGC
2473

omega-3 fatty acid
GAGCTCGGTTTTATGAGAATGTGGTTTTAGACCTCTCCCCAGATTC

desaturase
TACCCTAAACACACAACCTCTTTTGCCTC

Arabidopsis thaliana

GAGGCAAAAGAGGTTGTGTGTTTAGGGTAGAATCTGGGGAGAGGT
2474

Ser7 Term
CTAAAACCACATTCTCATAAAACCGAGCTCGCCATTGGAGCCTCTT

TCA-TGA
CCAAGAAGAAAAGAGGAAAAAGTCTCTGT

GGTTTTATGAGAATGTG
2475

CACATTCTCATAAAACC
2476

Reducing linolenic acid
AGAGACTTTTTCCTCTTTTCTTCTTGGGAAGAGGCTCCAATGGCGA
2477

omega-3 fatty acid
GCTCGGTTTTATCATAATGTGGTTTTAGACCTCTCCCCAGATTCTA

desaturase
CCCTAAACACACAACCTCTTTTGCCTCTA

Arabidopsis thaliana

TAGAGGCAAAAGAGGTTGTGTGTTTAGGGTAGAATCTGGGGAGAG
2478

Glu8 Term
GTCTAAAACCACATTATGATAAAACCGAGCTCGCCATTGGAGCCTC

GAA-TAA
TTCCCAAGAAGAAAAGAGGAAAAAGTCTCT

TTTTATCATAATGTGGT
2479

ACCACATTATGATAAAA
2480

Reducing linolenic acid
TCATCATCTTCTTCTTCTGGGGAGAGAGAGAGAGCAAAAGAGCTC
2481

omega-3 fatty acid
TAGCAATGGCGAACTAGGTCTTATCCGAATGTGGCATAAGACCTC

desaturase
TCCCCAGAATCTACACCACACCCAGATCCAC

Brassica juncea

GTGGATCTGGGTGTGGTGTAGATTCTGGGGAGAGGTCTTATGCCA
2482

Leu4 Term
CATTCGGATAAGACCTAGTTCGCCATTGCTAGAGCTCTTTTGCTCT

TTG-TAG
CTCTCTCTCCCCAGAAGAAGAAGATGATGA

GGCGAACTAGGTCTTAT
2483

ATAAGACCTAGTTCGCC
2484

Reducing linolenic acid
TCTTCTTCTTCTGGGGAGAGAGAGAGAGCAAAAGAGCTCTAGCAA
2485

omega-3 fatty acid
TGGCGAACTTGGTCTGATCCGAATGTGGCATAAGACCTCTCCCCA

desaturase
GAATCTACACCACACCCAGATCCACTTTCCT

Brassica juncea

AGGAAAGTGGATCTGGGTGTGGTGTAGATTCTGGGGAGAGGTCTT
2486

Leu6 Term
ATGCCACATTCGGATCAGACCAAGTTCGCCATTGCTAGAGCTCTTT

TTA-TGA
TGCTCTCTCTCTCTCCCCAGAAGAAGAAGA

CTTGGTCTGATCCGAAT
2487

ATTCGGATCAGACCAAG
2488

Reducing linolenic acid
TTCTTCTGGGGAGAGAGAGAGAGCAAAAGAGCTCTAGCAATGGCG
2489

omega-3 fatty acid
AACTTGGTCTTATCCTAATGTGGCATAAGACCTCTCCCCAGAATCT

desaturase
ACACCACACCCAGATCCACTTTCCTCTCCA

Brassica juncea

TGGAGAGGAAAGTGGATCTGGGTGTGGTGTAGATTCTGGGGAGA
2490

Glu8 Term
GGTCTTATGCCACATTAGGATAAGACCAAGTTCGCCATTGCTAGA

GAA-TAA
GCTCTTTTGCTCTCTCTCTCTCCCCAGAAGAA

TCTTATCCTAATGTGGC
2491

GCCACATTAGGATAAGA
2492

Reducing linolenic acid
CTGGGGAGAGAGAGAGAGCAAAAGAGCTCTAGCAATGGCGAACT
2493

omega-3 fatty acid
TGGTCTTATCCGAATGAGGCATAAGACCTCTCCCCAGAATCTACAC

desaturase
CACACCCAGATCCACTTTCCTCTCCAACACC

Brassica juncea

GGTGTTGGAGAGGAAAGTGGATCTGGGTGTGGTGTAGATTCTGG
2494

Cys9 Term
GGAGAGGTCTTATGCCTCATTCGGATAAGACCAAGTTCGCCATTG

TGT-TGA
CTAGAGCTCTTTTGCTCTCTCTCTCTCCCCAG

TCCGAATGAGGCATAAG
2495

CTTATGCCTCATTCGGA
2496

Reducing linolenic acid
ATAACAGAATTGCTGAATTCTTGCATTTTTAGCTTCTGGGTTTTCAA
2497

omega-3 fatty acid
TGGCTGCTGGTTGAGTATTATCAGAATGTGGTTTAAGGCCTCTCCC

desaturase
AAGAATCTACTCACGACCCAGAATTGGT

Ricinus communis

ACCAATTCTGGGTCGTGAGTAGATTCTTGGGAGAGGCCTTAAACC
2498

Trp5 Term
ACATTCTGATAATACTCAACCAGCAGCCATTGAAAACCCAGAAGCT

TGG-TGA
AAAAATGCAAGAATTCAGCAATTCTGTTAT

GCTGGTTGAGTATTATC
2499

GATAATACTCAACCAGC
2500

Reducing linolenic acid
AGAATTGCTGAATTCTTGCATTTTTAGCTTCTGGGTTTTCAATGGCT
2501

omega-3 fatty acid
GCTGGTTGGGTATGATCAGAATGTGGTTTAAGGCCTCTCCCAAGA

desaturase
ATCTACTCACGACCCAGAATTGGTTTTAC

Ricinus communis

GTAAAACCAATTCTGGGTCGTGAGTAGATTCTTGGGAGAGGCCTT
2502

Leu7 Term
AAACCACATTCTGATCATACCCAACCAGCAGCCATTGAAAACCCAG

TTA-TGA
AAGCTAAAAATGCAAGAATTCAGCAATTCT

TTGGGTATGATCAGAAT
2503

ATTCTGATCATACCCAA
2504

Reducing linolenic acid
ATTGCTGAATTCTTGCATTTTTAGCTTCTGGGTTTTCAATGGCTGCT
2505

omega-3 fatty acid
GGTTGGGTATTATGAGAATGTGGTTTAAGGCCTCTCCCAAGAATCT

desaturase
ACTCACGACCCAGAATTGGTTTTACATC

Ricinus communis

GATGTAAAACCAATTCTGGGTCGTGAGTAGATTCTTGGGAGAGGC
2506

Ser8 Term
CTTAAACCACATTCTCATAATACCCAACCAGCAGCCATTGAAAACC

TCA-TGA
CAGAAGCTAAAAATGCAAGAATTCAGCAAT

GGTATTATGAGAATGTG
2507

CACATTCTCATAATACC
2508

Reducing linolenic acid
TGCTGAATTCTTGCATTTTTAGCTTCTGGGTTTTCAATGGCTGCTG
2509

omega-3 fatty acid
GTTGGGTATTATCATAATGTGGTTTAAGGCCTCTCCCAAGAATCTA

desaturase
CTCACGACCCAGAATTGGTTTTACATCGA

Ricinus communis

TCGATGTAAAACCAATTCTGGGTCGTGAGTAGATTCTTGFGGAGAG
2510

Glu9 Term
CGCCTTAAACCACATTATGATAATACCCAACCAGCAGCCATTGAAAA

GAA-TAA
CCCAGAAGCTAAAAATGCAAGAATTCAGCA

TATTATCATAATGTGGT
2511

ACCACATTATGATAATA
2512

Reducing linolenic acid
GCAAGTTGGTTTTATCAGAATGTGGTCTTAGACCACTCCCAAGAA
2513

omega-3 fatty acid
TCTACCCTAAGCCCTGAACTGGGGCAGCCACTTCTGCCTCCTCTC

desaturase
ACATTAAGTTGAGAATTTCACGTACAGATC

Nicotiana tabacum

GATCTGTACGTGAAATTCTCAACTTAATGTGAGAGGAGGCAGAAGT
2514

Arg22 Term
GGCTGCCCCAGTTCAGGGCTTAGGGTAGFATTCTTGGGAGTGGTCT

AGA-TGA
AAGACCACATTCTGATAAAACCCAACTTGC

CTAAGCCCTGAACTGGG
2515

CCCAGTTCAGGGCTTAG
2516

Reducing linolenic acid
CTCCCAAGAATCTACCCTAAGCCCAGAACTGGGGCAGCCACTTCT
2517

omega-3 fatty acid
GCCTCCTCTCACATTTAGTTGAGAATTTCACGTACAGATCTGAGTG

desaturase
GTTCTGCAATTTCTTTGTCTAATACTAAT

Nicotiana tabacum

TATTAGTATTAGACAAAGAAATTGCAGAACCACTCAGATCTGTACG
2518

Lys34 Term
TGAAATTCTCAACTAAATGTGAGAGGAGGCAGAAGTGGCTGCCCC

AAG-TAG
AGTTCTGGGCTTAGGGTAGATTCTTGGGAG

CTCACATTTAGTTGAGA
2519

TCTCAACTAAATGTGAG
2520

Reducing linolenic acid
CAAGAATCTACCCTAAGCCCAGAACTGGGGCAGCCACTTCTGCCT
2521

omega-3 fatty acid
CCTCTCACATTAAGTAGAGAATTTCACGTACAGATCTGAGTGGTTC

desaturase
TGCAATTTCTTTGTCTAATACTAATAAAGA

Nicotiana tabacum

TCTTTATTAGTATTAGACAAAGAAATTGCAGAACCACTCAGATCTGT
2522

Leu35 Term
ACGTGAAATTCTCTACTTAATGTGAGAGGAGGCAGAAGTGGCTGC

TTG-TAG
CCCAGTTCTGGGCTTAGGGTAGATTCTTG

CATTAAGTAGAGAATTT
2523

AAATTCTCTACTTAATG
2524

Reducing linolenic acid
AGAATCTACCCTAAGCCCAGAACTGGGGCAGCCACTTCTGCCTCC
2525

omega-3 fatty acid
TCTCACATTAAGTTGTGAATTTCACGTACAGATCTGAGTGGTTCTG

desaturase
CAATTTCTTTGTCTAATACTAATAAAGAGA

Nicotiana tabacum

TCTCTTTATTAGTATTAGACAAAGAAATTGCAGAACCACTCAGATCT
2526

Arg36 Term
GTACGTGAAATTCACAACTTAATGTGAGAGGAGGCAGAAGTGGCT

AGA-TGA
GCCCCAGTTCTGGGCTTAGGGTAGATTCT

TTAAGTTGTGAATTTCA
2527

TGAAATTCACAACTTAA
2528

Reducing linolenic acid
GCGAGTTGGGTTTTATCAGAATGTGGTCTGAGGCCACTCCCGAGG
2529

omega-3 fatty acid
GTCTATCCTAAGCCATGAACTGGCCACCCTTTGTTGAATTCCAATC

desaturase
CCACAAAGCTGAGATTTTCAAGAACAGATC

Sesamum indicum

GATCTGTTCTTGAAAATCTCAGCTTTGTGGGATTGGAATTCAACAA
2530

Arg22 Term
AGGGTGGCCAGTTCATGGCTTAGGATAGACCCTCGGGAGTGGCC

AGA-TGA
TCAGACCACATTCTGATAAAACCCAACTCGC

CTAAGCCATGAACTGGC
2531

GCCAGTTCATGGCTTAG
2532

Reducing linolenic acid
CAGAATGTGGTCTGAGGCCACTCCCGAGGGTCTATCCTAAGCCAA
2533

omega-3 fatty acid
GAACTGGCCACCCTTAGTTGAATTCCAATCCCACAAAGCTGAGATT

desaturase
TTCAAGAACAGATCTTGGAAATGGTTCTTC

Sesamum indicum

GAAGAACCATTTCCAAGATCTGTTCTTGAAAATCTCAGCTTTGTGG
2534

Leu27 Term
GATTGGAATTCAACTAAGGGTGGCCAGTTCTTGGCTTAGGATAGA

TTG-TAG
CCCTCGGGAGTGGCCTCAGACCACATTCTG

CCACCCTTAGTTGAATT
2535

AATTCAACTAAGGGTGG
2536

Reducing linolenic acid
AATGTGGTCTGAGGCCACTCCCGAGGGTCTATCCTAAGCCAAGAA
2537

omega-3 fatty acid
CTGGCCACCCTTTGTAGAATTCCAATCCCACAAAGCTGAGATTTTC

desaturase
AAGAACAGATCTTGGAAATGGTTCTTCATT

Sesamum indicum

AATGAAGAACCATTTCCAAGATCTGTTCTTGAAAATCTCAGCTTTGT
2538

Leu28 Term
GGGATTGGAATTCTACAAAGGGTGGCCAGTTCTTGGCTTAGGATA

TTG-TAG
GACCCTCGGGAGTGGCCTCAGACCACATT

CCCTTTGTAGAATTCCA
2539

TGGAATTCTACAAAGGG
2540

Reducing linolenic acid
CTCCCGAGGGTCTATCCTAAGCCAAGAACTGGCCACCCTTTGTTG
2541

omega-3 fatty acid
AATTCCAATCCCACATAGCTGAGATTTTCAAGAACAGATCTTGGAA

desaturase
ATGGTTCTTCATTCTGTTTGTCGAGTGGGA

Sesamum indicum

TCCCACTCGACAAACAGAATGAAGAACCATTTCCAAGATCTGTTCT
2542

Lys34 Term
TGAAAATCTCAGCTATGTGGGATTGGAATTCAACAAAGGGTGGCC

AAG-TAG
AGTTCTTGGCTTAGGATAGACCCTCGGGAG

ATCCCACATAGCTGAGA
2543

TCTCAGCTATGTGGGAT
2544

Reducing linolenic acid
CATCAGAGCGGCGATACCTAAGCATTGCTGGGTTAAGAATCCATG
2545

omega-3 fatty acid
GAAGTCTATGAGTTAGGTCGTCAGAGAGCTAGCCATCGTGTTCGC

desaturase
ACTAGCTGCTGGAGCTGCTTACCTCAACAAT

Brassica napus

ATTGTTGAGGTAAGCAGCTCCAGCAGCTAGTGCGAACACGATGGC
2546

Tyr3 Term
TAGCTCTCTGACGACCTAACTCATAGACTTCCATGGATTCTTAACC

TAC-TAG
CAGCAATGCTTAGGTATCGCCGCTCTGATG

ATGAGTTAGGTCGTCAG
2547

CTGACGACCTAACTCAT
2548

Reducing linolenic acid
GCGGCGATACCTAAGCATTGCTGGGTTAAGAATCCATGGAAGTCT
2549

omega-3 fatty acid
ATGAGTTACGTCGTCTGAGAGCTAGCCATCGTGTTCGCACTAGCT

desaturase
GCTGGAGCTGCTTACCTCAACAATTGGCTTG

Brassica napus

CAAGCCAATTGTTGAGGTAAGCAGCTCCAGCAGCTAGTGCGAACA
2550

Arg6 Term
CGATGGCTAGCTCTCAGACGACGTAACTCATAGACTTCCATGGAT

AGA-TGA
CTTAACCCAGCAATGCTTAGGTATCGCCGC

ACGTCGTCTGAGAGCTA
2551

TAGCTCTCAGACGACGT
2552

Reducing linolenic acid
GCGATACCTAAGCATTGCTGGGTTAAGAATCCATGGAAGTCTATGA
2553

omega-3 fatty acid
GTTACGTCGTCAGATAGCTAGCCATCGTGTTCGCACTAGCTGCTG

desaturase
GAGCTGCTTACCTCAACAATTGGCTTGTTT

Brassica napus

AAACAAGCCAATTGTTGAGGTAAGCAGCTCCAGCAGCTAGTGCGA
2554

Glu7 Term
ACACGATGGCTAGCTATCTGACGACGTAACTCATAGACTTCCATG

GAG-TAG
GATTCTTAACCCAGCAATGCTTAGGTATCGC

TCGTCAGATAGCTAGCC
2555

GGCTAGCTATCTGACGA
2556

Reducing linolenic acid
CCATGGAAGTCTATGAGTTACGTCGTCAGAGAGCTAGCCATCGTG
2557

omega-3 fatty acid
TTCGCACTAGCTGCTTGAGCTGCTTACCTCAACAATTGGCTTGTTT

desaturase
GGCCTCTCTATTGGATTGCTCAAGGAACCA

Brassica napus

TGGTTCCTTGAGCAATCCAATAGAGAGGCCAAACAAGCCAATTGTT
2558

Gly17 Term
GAGGTAAGCAGCTCAAGCAGCTAGTGCGAACACGATGGCTAGCT

GGA-TGA
CTCTGACGACGTAACTCATAGACTTCCATGG

TAGCTGCTTGAGCTGCT
2559

AGCAGCTCAAGCAGCTA
2560

Reducing linolenic acid
GCAAGTTGGGTTCTATCAGAATGTGGTCTTAGACCACTACCAAGAA
2561

omega-3 fatty acid
TATACCCAAAGCCCTGAATAGGGTCTTCTTCCGTTTGCGCCACCAA

desaturase
TTTAAATCTGAGAAGAATTTCACCTTCAC

Solanum tuberosum

GTGAAGGTGAAATTCTTCTCAGATTTAAATTGGTGGCGCAAACGGA
2562

Arg22 Term
AGAAGACCCTATTCAGGGCTTTGGGTATATTCTTGGTAGTGGTCTA

AGA-TGA
AGACCACATTCTGATAGAACCCAACTTGC

CAAAGCCCTGAATAGGG
2563

CCCTATTCAGGGCTTTG
2564

Reducing linolenic acid
TGGTCTTAGACCACTACCAAGAATATACCCAAAGCCCAGAATAGG
2565

omega-3 fatty acid
GTCTTCTTCCGTTTGAGCCACCAATTTAAATCTGAGAAGAATTTCA

desaturase
CCTTCACCTATACGAACAGATCGGAATTGT

Solanum tuberosum

ACAATTCCGATCTGTTCGTATAGGTGAAGGTGAAATTCTTCTCAGA
2566

Cys29 Term
TTTAAATTGGTGGCTCAAACGGAAGAAGACCCTATTCTGGGCTTTG

TGC-TGA
GGTATATTCTTGGTAGTGGTCTAAGACCA

TCCGTTTGAGCCACCAA
2567

TTGGTGGCTCAAACGGA
2568

Reducing linolenic acid
CACTACCAAGAATATACCCAAAGCCCAGAATAGGGTCTTCTTCCGT
2569

omega-3 fatty acid
TTGCGCCACCAATTGAAATCTGAGAAGAATTTCACCTTCACCTATA

desaturase
CGAACAGATCGGAATTGTTGGGCATTGAG

Solanum tuberosum

CTCAATGCCCAACAATTCCGATCTGTTCGTATAGGTGAAGGTGAAA
2570

Leu33 Term
TTCTTCTCAGATTTCAATTGGTGGCGCAAACGGAAGAAGACCCTAT

TTA-TGA
TCTGGGTTTGGGTATATTCTTGGTAGTG

CACCAATTGAAATCTGA
2571

TCAGATTTCAATTGGTG
2572

Reducing linolenic acid
AGAATATACCCAAAGCCCAGAATAGGGTCTTCTTCCGTTTGCGCCA
2573

omega-3 fatty acid
CCAATTTAAATCTGTGAAGAATTTCACCTTCACCTATACGAACAGAT

desaturase
CGGAATTGTTGGGCATTGAGGGTAAGTG

Solanum tuberosum

CACTTACCCTCAATGCCCAACAATTCCGATCTGTTCGTATAGGTGA
2574

Arg36 Term
AGGTGAAATTCTTCACAGATTTAAATTGGTGGCGCAAACGGAAGAA

AGA-TGA
GACCCTATTCTGGGCTTTGGGTATATTCT

TAAATCTGTGAAGAATT
2575

AATTCTTCACAGATTTA
2576

Reducing linolenic acid
CTCTTTATTATCCTCCTCTTCTTTGTTTTTTTTGAGTTCTGAGTCACC
2577

omega-3 fatty acid
TATGGCAAGTTGAGTGATTTCAGAATGTGGGCTAAGGCCACTTCC

desaturase
AAGAATCTATGCCAGGCCCAGAAGTGGA

Petroselinum crispum

TCCACTTCTGGGCCTGGCATAGATTCTTGGAAGTGGCCTTAGCCC
2578

Trp4 Term
ACATTCTGAAATCACTCAACTTGCCATAGGTGACTCAGAACTCAAA

TGG-TGA
AAAAACAAAGAAGAGGAGGATAATAAAGAG

GCAAGTTGAGTGATTTC
2579

GAAATCACTCAACTTGC
2580

Reducing linolenic acid
TATCCTCCTCTTCTTTGTTTTTTTTGAGTTCTGAGTCACCTATGGCA
2581

omega-3 fatty acid
AGTTGGGTGATTTGAGAATGTGGGCTAAGGCCACTTCCAAGAATC

desaturase
TATGCCAGGCCCAGAAGTGGAGCTTCATG

Petroselinum crispum

CATGAAGCTCCACTTCTGGGCCTGGCATAGATTCTTGGAAGTGGC
2582

Ser7 Term
CTTAGCCCACATTCTCAAATCACCCAACTTGCCATAGGTGACTCAG

TCA-TGA
AACTCAAAAAAAACAAAGAAGAGGAGGATA

GGTGATTTGAGAATGTG
2583

CACATTCTCAAATCACC
2584

Reducing linolenic acid
TCCTCCTCTTCTTTGTTTTTTTTGAGTTCTGAGTCACCTATGGCAAG
2585

omega-3 fatty acid
TTGGGTGATTTCATAATGTGGGCTAAGGCCACTTCCAAGAATCTAT

desaturase
GCCAGGCCCAGAAGTGGAGCTTCATGTT

Petroselinum crispum

AACATGAAGCTCCACTTCTGGGCCTGGCATAGATTCTTGGAAGTG
2586

Glu8 Term
GCCTTAGCCCACATTATGAAATCACCCAACTTGCCATAGGTGACTC

GAA-TAA
AGAACTCAAAAAAAACAAAGAAGAGGAGGA

TGATTTCATAATGTGGG
2587

CCCACATTATGAAATCA
2588

Reducing linolenic acid
CTCTTCTTTGTTTTTTTTGAGTTCTGAGTCACCTATGGCAAGTTGGG
2589

omega-3 fatty acid
TGATTTCAGAATGAGGGCTAAGGCCACTTCCAAGAATCTATGCCA

desaturase
GGCCCAGAAGTGGAGCTTCATGTTTCAAC

Petroselinum crispum

GTTGAAACATGAAGCTCCACTTCTGGGCCTGGCATAGATTCTTGG
2590

Cys9 Term
AAGTGGCCTTAGCCCTCATTCTGAAATCACCCAACTTGCCATAGGT

TGT-TGA
GACTCAGAACTCAAAAAAAACAAAGAAGAG

TCAGAATGAGGGCTAAG
2591

CTTAGCCCTCATTCTGA
2592

Reducing linolenic acid
ATGAAGCAGCAACAGTACAAAGACACCCCAATTCTAAATGGCGTTA
2593

omega-3 fatty acid
ATGGTTTTCATGCTTAAGAAGAAGAAGAAGAAGAGGATTTCGACTT

desaturase
AAGCAATCCTCCTCCATTCAATATTGGTC

Vernicia fordii

GACCAATATTGAATGGAGGAGGATTGCTTAAGTCGAAATCCTCTTC
2594

Lys21 Term
TTCTTCTTCTTCTTAAGCATGAAAACCATTAACGCCATTTAGAATTG

AAA-TAA
GGGTGTCTTTGTACTGTTGCTGCTTCAT

TTCATGCTTAAGAAGAA
2595

TTCTTCTTAAGCATGAA
2596

Reducing linolenic acid
AAGCAGCAACAGTACAAAGACACCCCAATTCTAAATGGCGTTAATG
2597

omega-3 fatty acid
GTTTTCATGCTAAATAAGAAGAAGAAGAAGAGGATTTCGACTTAAG

desaturase
CAATCCTCCTCCATTCAATATTGGTCAGA

Vernicia fordii

TCTGACCAATATTGAATGGAGGAGGATTGCTTAAGTCGAAATCCTC
2598

Glu22 Term
TTCTTCTTCTTCTTATTTAGCATGAAAACCATTAACGCCATTTAGAA

GAA-TAA
TTGGGGTGTCTTTGTACTGTTGCTGCTT

ATGCTAAATAAGAAGAA
2599

TTCTTCTTATTTAGCAT
2600

Reducing linolenic acid
CAGCAACAGTACAAAGACACCCCAATTCTAAATGGCGTTAATGGTT
2601

omega-3 fatty acid
TTCATGCTAAAGAATAAGAAGAAGAAGAGGATTTCGACTTAAGCAA

desaturase
TCCTCCTCCATTCAATATTGGTCAGATCC

Vernicia fordii

GGATCTGACCAATATTGAATGGAGGAGGATTGCTTAAGTCGAAATC
2602

Glu23 Term
CTCTTCTTCTTCTTATTCTTTAGCATGAAAACCATTAACGCCATTTA

GAA-TAA
GAATTGGGGTGTCTTTGTACTGTTGCTG

CTAAAGAATAAGAAGAA
2603

TTCTTCTTATTCTTTAG
2604

Reducing linolenic acid
CAGCAACAGTACAAAGACACCCCAATTCTAAATGGCGTTAATGGTT
2605

omega-3 fatty acid
TTCATGCTAAAGAATAAGAAGAAGAAGAGGATTTCGACTTAAGCAA

desaturase
TCCTCCTCCATTCAATATTGGTCAGATCC

Vernicia fordii

GGATCTGACCAATATTGAATGGAGGAGGATTGCTTAAGTCGAAATC
2606

Glu24 Term
CTCTTCTTCTTCTTATTCTTTAGCATGAAAACCATTAACGCCATTTA

GAA-TAA
GAATTGGGGTGTCTTTGTACTGTTGCTG

CTAAAGAATAAGAAGAA
2607

TTCTTCTTATTCTTTAG
2608

Reducing linolenic acid
GGTCCAAGCACAGCCTCTACAACATGTTGGTAATGGTGCAGGGAA
2609

omega-3 fatty acid
AGAAGATCAAGCTTAGTTTGATCCAAGTGCTCCACCACCCTTCAAG

desaturase
ATTGCAAATATCAGAGCAGCAATTCCAAAA

Glycine max

TTTTGGAATTGCTGCTCTGATATTTGCAATCTTGAAGGGTGGTGGA
2610

Tyr21 Term
GCACTTGGATCAAACTAAGCTTGATCTTCTTTCCCTGCACCATTAC

TAT-TAG
CAACATGTTGTAGAGGCTGTGCTTGGACC

CAAGCTTAGTTTGATCC
2611

GGATCAAACTAAGCCTG
2612

Reducing linolenic acid
GGTAATGGTGCAGGGAAAGAAGATCAAGCTTATTTTGATCCAAGT
2613

omega-3 fatty acid
GCTCCACCACCCTTCTAGATTGCAAATATCAGAGCAGCAATTCCAA

desaturase
AACATTGCTGGGAGAAGAACACATTGAGAT

Glycine max

ATCTCAATGTGTTCTTCTCCCAGCAATGTTTTGGAATTGCTGCTCT
2614

Lys31 Term
GATATTTGCAATCTAGAAGGGTGGTGGAGCACTTGGATCAAAATAA

AAG-TAG
GCTTGATCTTCTTTCCCTGCACCATTACC

CACCCTTCTAGATTGCA
2615

TGCAATCTAGAAGGGTG
2616

Reducing linolenic acid
AAAGAAGATCAAGCTTATTTTGATCCAAGTGCTCCACCACCCTTCA
2617

omega-3 fatty acid
AGATTGCAAATATCTGAGCAGCAATTCCAAAACATTGCTGGGAGAA

desaturase
GAACACATTGAGATCTCTGAGTTATGTTC

Glycine max

GAACATAACTCAGAGATCTCAATGTGTTCTTCTCCCAGCAATGTTTT
2618

Arg36 Term
GGAATTGCTGCTCAGATATTTGCAATCTTGAAGGGTGGTGGAGCA

AGA-TGA
CTTGGATCAAAATAAGCTTGATCTTCTTT

CAAATATCTGAGCAGCA
2619

TGCTGCTCAGATATTTG
2620

Reducing linolenic acid
TATTTTGATCCAAGTGCTCCACCACCCTTCAAGATTGCAAATATCA
2621

omega-3 fatty acid
GAGCAGCAATTCCATAACATTGCTGGGAGAAGAACACATTGAGAT

desaturase
CTCTGAGTTATGTTCTGAGGGATGTGTTGG

Glycine max

CCAACACATCCCTCAGAACATAACTCAGAGATCTCAATGTGTTCTT
2622

Leu41 Term
CTCCCAGCAATGTTATGGAATTGCTGCTCTGATATTTGCAATCTTG

AAA-TAA
AAGGGTGGTGGAGCACTTGGATCAAAATA

CAATTCCATAACATTGC
2623

GCAATGTTATGGAATTG
2624

Reducing linolenic acid
CATCCACCCGCACCCGCACCCGCCCCGCTGACGGCGGCAATGGC
2625

omega-3 fatty acid
CCGGCTCGTGCTCTCCTAGTGCTCGGGCCTCGCGCCCGTCCGCC

desaturase
GCCTGCGCGCCGGCCGGGGCGCCATTGCGGCGC

Zea mays

GCGCCGCAATGGCGCCCCGGCCGGCGCGCAGGCGGCGGACGG
2626

Glu8 Term
GCGCGAGGCCCGAGCACTAGGAGAGCACGAGCCGGGCCATTGC

GAG-TAG
CGCCGTCAGCGGGGCGGGTGCGGGTGCGGGTGGATG

TGCTCTCCTAGTGCTCG
2627

CGAGCACTAGGAGAGCA
2628

Reducing linolenic acid
ACCCGCACCCGCACCCGCCCCGCTGACGGCGGCAATGGCCCGG
2629

omega-3 fatty acid
CTCGTGCTCTCCGAGTGATCGGGCCTCGCGCCCGTCCGCCGCCT

desaturase
GCGCGCCGGCCGGGGCGCCATTGCGGCGCGGTCA

Zea mays

TGACCGCGCCGCAATGGCGCCCCGGCCGGCGCGCAGGCGGCGG
2630

Cys9 Term
ACGGGCGCGAGGCCCGATCACTCGGAGAGCACGAGCCGGGCCA

TGC-TGA
TTGCCGCCGTCAGCGGGGCGGGTGCGGGTGCGGGT

TCCGAGTGATCGGGCCT
2631

AGGCCCGATCACTCGGA
2632

Reducing linolenic acid
CCGCACCCGCACCCGCCCCGCTGACGGCGGCAATGGCCCGGCT
2633

omega-3 fatty acid
CGTGCTCTCCGAGTGCTAGGGCCTCGCGCCCGTCCGCCGCCTGC

desaturase
GCGCCGGCCGGGGCGCCATTGCGGCGCGGTCACC

Zea mays

GGTGACCGCGCCGCAATGGCGCCCCGGCCGGCGCGCAGGCGGC
2634

Ser10 Term
GGACGGGCGCGAGGCCCTAGCACTCGGAGAGCACGAGCCGGGC

TCG-TAG
CATTGCCGCCGTCAGCGGGGCGGGTGCGGGTGCGG

CGAGTGCTAGGGCCTCG
2635

CGAGGCCCTAGCACTCG
2636

Reducing linolenic acid
GCTCGGGCCTCGCGCCCGTCCGCCGCCTGCGCGCCGGCCGGGG
2637

omega-3 fatty acid
CGCCATTGCGGCGCGGTGACCCCCCGCGCTCTCCGCGGCGCCG

desaturase
CGCCGTCGTCCCGCGTCCGCGTCCATCCACCGCGA

Zea mays

TCGCGGTGGATGGACGCGGACGCGGGACGACGGCGCGGCGCCG
2638

Ser29 Term
CGGAGAGCGCGGGGGGTCACCGCGCCGCAATGGCGCCCCGGCC

TCA-TGA
GGCGCGCAGGCGGCGGACGGGCGCGAGGCCCGAGC

GGCGCGGTGACCCCCCG
2639

CGGGGGGTCACCGCGCC
2640

Reducing linolenic acid
CCCCCTCCCCCACGCACACGCACAGATCCATCCGCGGCCATGGC
2641

omega-3 fatty acid
CCCCGCAATGAGGCCGTAGCAGGAGGCGAGCTGCAAGGCCACCG

desaturase
AGGACCACCGCTCCGAGTTCGACGCCGCCAAGC

Triticum aestivum

GCTTGGCGGCGTCGAACTCGGAGCGGTGGTCCTCGGTGGCCTTG
2642

Glu8 Term
CAGCTCGCCTCCTGCTACGGCCTCATTGCGGGGGCCATGGCCGC

GAG-TAG
GGATGGATCTGTGCGTGTGCGTGGGGGAGGGGG

TGAGGCCGTAGCAGGAG
2643

CTCCTGCTACGGCCTCA
2644

Reducing linolenic acid
CCTCCCCCACGCACACGCACAGATCCATCCGCGGCCATGGCCCC
2645

omega-3 fatty acid
CGCAATGAGGCCGGAGTAGGAGGCGAGCTGCAAGGCCACCGAG

desaturase
GACCACCGCTCCGAGTTCGACGCCGCCAAGCCGC

Triticum aestivum

GCGGCTTGGCGGCGTCGAACTCGGAGCGGTGGTCCTCGGTGGCC
2646

Gln9 Term
TTGCAGCTCGCCTCCTACTCCGGCCTCATTGCGGGGGCCATGGC

CAG-TAG
CGCGGATGGATCTGTGCGTGTGCGTGGGGGAGG

GGCCGGAGTAGGAGGCG
2647

CGCCTCCTACTCCGGCC
2648

Reducing linolenic acid
CCCCCACGCACACGCACAGATCCATCCGCGGCCATGGCCCCCGC
2649

omega-3 fatty acid
AATGAGGCCGGAGCAGTAGGCGAGCTGCAAGGCCACCGAGGACC

desaturase
ACCGCTCCGAGTTCGACGCCGCCAAGCCGCCGC

Triticum aestivum

GCGGCGGCTTGGCGGCGTCGAACTCGGAGCGGTGGTCCTCGGT
2650

Glu10 Term
GGCCTTGCAGCTCGCCTACTGCTCCGGCCTCATTGCGGGGGCCA

GAG-TAG
TGGCCGCGGATGGATCTGTGCGTGTGCGTGGGGG

CGGAGCAGTAGGCGAGC
2651

GCTCGCCTACTGCTCCG
2652

Reducing linolenic acid
ACGCACAGATCCATCCGCGGCCATGGCCCCCGCAATGAGGCCGG
2653

omega-3 fatty acid
AGCAGGAGGCGAGCTGAAAGGCCACCGAGGACCACCGCTCCGA

desaturase
GTTCGACGCCGCCAAGCCGCCGCCCTTCCGCATC

Triticum aestivum

GATGCGGAAGGGCGGCGGCTTGGCGGCGTCGAACTCGGAGCGG
2654

Cys13 TermTGGTCCTCGGTGGCCTTTCAGCTCGCCTCCTGCTCCGGCCTCATT

TGC-TGA
GCGGGGGCCATGGCCGCGGATGGATCTGTGCGT

GCGAGCTGAAAGGCCAC
2655

GTGGCCTTTCAGCTCGC
2656

Reducing linolenic acid
CTTCACAAATCACAAATCGGAATCAGATCCACCACGACACCCCGG
2657

omega-3 fatty acid
CGGCAATGGCGGCGTAGGCGACCCAGGAGGCCGACTGCAAGGC

desaturase
TTCCGAGGACGCCCGTCTCTTCTTCGACGCCGC

Oryza sativa

GCGGCGTCGAAGAAGAGACGGGCGTCCTCGGAAGCCTTGCAGTC
2658

Ser4 Term
GGCCTCCTGGGTCGCCTACGCCGCCATTGCCGCCGGGGTGTCGT

TCG-TAG
GGTGGATCTGATTCCGATTTGTGATTTGTGAAG

GGCGGCGTAGGCGACCC
2659

GGGTCGCCTACGCCGCC
2660

Reducing linolenic acid
ATCACAAATCGGAATCAGATCCACCACGACACCCCGGCGGCAATG
2661

omega-3 fatty acid
GCGGCGTCGGCGACCTAGGAGGCCGACTGCAAGGCTTCCGAGGA

desaturase
CGCCCGTCTCTTCTTCGACGCCGCCAAGCCCC

Oryza sativa

GGGGCTTGGCGGCGTCGAAGAAGAGACGGGCGTCCTCGGAAGC
2662

Gln7 Term
CTTGCAGTCGGCCTCCTAGGTCGCCGACGCCGCCATTGCCGCCG

CAG-TAG
GGGTGTCGTGGTGGATCTGATTCCGATTTGTGAT

CGGCGACCTAGGAGGCC
2663

GGCCTCCTAGGTCGCCG
2664

Reducing linolenic acid
ACAAATCGGAATCAGATCCACCACGACACCCCGGCGGCAATGGC
2665

omega-3 fatty acid
GGCGTCGGCGACCCAGTAGGCCGACTGCAAGGCTTCCGAGGACG

desaturase
CCCGTCTCTTCTTCGACGCCGCCAAGCCCCCGC

Oryza sativa

GCGGGGGCTTGGCGGCGTCGAAGAAGAGACGGGCGTCCTCGGA
2666

Glu8 Term
AGCCTTGCAGTCGGCCTACTGGGTCGCCGACGCCGCCATTGCCG

GAG-TAG
CCGGGGTGTCGTGGTGGATCTGATTCCGATTTGT

CGACCCAGTAGGCCGAC
2667

GTCGGCCTACTGGGTCG
2668

Reducing linolenic acid
TCAGATCCACCACGACACCCCGGCGGCAATGGCGGCGTCGGCGA
2669

omega-3 fatty acid
CCCAGGAGGCCGACTGAAAGGCTTCCGAGGACGCCCGTCTCTTC

desaturase
TTCGACGCCGCCAAGCCCCCGCCCTTCCGCATC

Oryza sativa

GATGCGGAAGGGCGGGGGCTTGGCGGCGTCGAAGAAGAGACGG
2670

Cys10 Term
GCGTCCTCGGAAGCCTTTCAGTCGGCCTCCTGGGTCGCCGACGC

TGC-TGA
CGCCATTGCCGCCGGGGTGTCGTGGTGGATCTGA

GCCGACTGAAAGGCTTC
2671

	Number	Date	Country
	60244989	Oct 2000	US
	60208538	Jun 2000	US

	Number	Date	Country
Parent	PCT/US01/17672	Jun 2001	US
Child	10307005	Nov 2002	US

	Number	Date	Country
Parent	09818875	Mar 2001	US
Child	PCT/US01/17672	Jun 2001	US

Targeted chromosomal genomic alterations in plants using modified single stranded oligonucleotides

Information

Publication Number

Date Filed

Date Published

Inventors

CPC

US Classifications

International Classifications

Abstract

Description

Claims

Provisional Applications (2)

Continuations (1)

Continuation in Parts (1)