Patent Grant
10806146
A computer readable form of a sequence listing is filed with this application by electronic submission and is incorporated into this application by reference in its entirety. The sequence listing is contained in the file named P34113US02_SEQ.txt, which is 849,269 bytes in size (measured in operating system MS windows) and was created on Jul. 28, 2016.
The methods and compositions generally relate to the field of weed management. More specifically, related to glutamine synthetase (GS) genes in plants and compositions containing polynucleotide molecules for modulating their expression. Further provided are methods and compositions useful for weed control.
Weeds are plants that compete with cultivated plants in an agronomic environment and cost farmers billions of dollars annually in crop losses and the expense of efforts to keep weeds under control. Weeds also serve as hosts for crop diseases and insect pests. The losses caused by weeds in agricultural production environments include decreases in crop yield, reduced crop quality, increased irrigation costs, increased harvesting costs, reduced land value, injury to livestock, and crop damage from insects and diseases harbored by the weeds. The principal means by which weeds cause these effects are: 1) competing with crop plants for water, nutrients, sunlight and other essentials for growth and development, 2) production of toxic or irritant chemicals that cause human or animal health problem, 3) production of immense quantities of seed or vegetative reproductive parts or both that contaminate agricultural products and perpetuate the species in agricultural lands, and 4) production on agricultural and nonagricultural lands of vast amounts of vegetation that must be disposed of Herbicide tolerant weeds are a problem with nearly all herbicides in use, there is a need to effectively manage these weeds. There are over 365 weed biotypes currently identified as being herbicide resistant to one or more herbicides by the Herbicide Resistance Action Committee (HRAC), the North American Herbicide Resistance Action Committee (NAHRAC), and the Weed Science Society of America (WSSA).
The glutamine synthetase (GS) enzyme is an essential enzyme in the metabolism of nitrogen by catalyzing the condensation of glutamate and ammonia to form glutamine. This enzyme is the target of phosphinic acids herbicides that include glufosinate-ammonium and bialaphos.
In one aspect, the invention provides a method of plant control comprising an external application to a plant of a composition comprising a polynucleotide and a transfer agent, wherein the polynucleotide is essentially identical or essentially complementary to a glutamine synthetase (GS) gene sequence or fragment thereof, or to the RNA transcript of said GS gene sequence or fragment thereof, wherein said GS gene sequence is selected from the group consisting of SEQ ID NOs: 1-59 or a polynucleotide fragment thereof, whereby the weedy plant growth or development or reproductive ability is reduced or the weedy plant is made more sensitive to a GS inhibitor herbicide relative to a weedy plant not treated with said composition. In this manner, plants that have become resistant to the application of GS inhibitor containing herbicides may be made more susceptible to the herbicidal effects of a GS inhibitor containing herbicide, thus potentiating the effect of the herbicide. The polynucleotide fragment is at least 18 contiguous nucleotides, at least 19 contiguous nucleotides, at least 20 contiguous nucleotides or at least 21 contiguous nucleotides in length and at least 85 percent identical to a GS gene sequence selected from the group consisting of SEQ ID NOs: 1-59 and the transfer agent is an organosilicone composition or compound. The polynucleotide fragment can also be sense or anti-sense ssDNA or ssRNA, dsRNA, or dsDNA, or dsDNA/RNA hybrids. The composition can include more than one polynucleotide fragments, and the composition can include a GS inhibitor herbicide and/or other herbicides (co-herbicides) that enhance the weed control activity of the composition.
In another aspect, polynucleotide molecules and methods for modulating GS gene expression in plant species are provided. The method reduces, represses or otherwise delays expression of a GS gene in a plant comprising an external application to a plant of a composition comprising a polynucleotide and a transfer agent, wherein the polynucleotide is essentially identical or essentially complementary to a GS gene sequence or fragment thereof, or to the RNA transcript of the GS gene sequence or fragment thereof, wherein the GS gene sequence is selected from the group consisting of SEQ ID NOs: 1-59 or a polynucleotide fragment thereof. The polynucleotide fragment is at least 18 contiguous nucleotides, at least 19 contiguous nucleotides, at least 20 contiguous nucleotides at least 21 contiguous nucleotides in length and at least 85 percent identical to a GS gene sequence selected from the group consisting of SEQ ID NOs:1-59 and the transfer agent is an organosilicone compound. The polynucleotide fragment can also be sense or anti-sense ssDNA or ssRNA, dsRNA, or dsDNA, or dsDNA/RNA hybrids. Polynucleotide molecules comprising SEQ ID NOs: 37-1056 are fragments of the GS gene.
In a further aspect, the polynucleotide molecule containing composition may be combined with other herbicidal (co-herbicides) compounds to provide additional control of unwanted plants in a field of cultivated plants.
In a further aspect, the polynucleotide molecule composition may be combined with any one or more additional agricultural chemicals, such as, insecticides, fungicides, nematicides, bactericides, acaricides, growth regulators, chemosterilants, semiochemicals, repellents, attractants, pheromones, feeding stimulants, biopesticides, microbial pesticides or other biologically active compounds to form a multi-component pesticide giving an even broader spectrum of agricultural protection.
Provided are methods and compositions containing a polynucleotide that provide for regulation, repression or delay of GS (glutamine synthetase) gene expression and enhanced control of weedy plant species and importantly GS inhibitor resistant weed biotypes. Aspects of the method can be applied to manage various weedy plants in agronomic and other cultivated environments.
The following definitions and methods are provided to better define the present invention and to guide those of ordinary skill in the art in the practice of the present invention. Unless otherwise noted, terms are to be understood according to conventional usage by those of ordinary skill in the relevant art. Where a term is provided in the singular, the inventors also contemplate aspects of the invention described by the plural of that term.
By “non-transcribable” polynucleotides is meant that the polynucleotides do not comprise a complete polymerase II transcription unit. As used herein “solution” refers to homogeneous mixtures and non-homogeneous mixtures such as suspensions, colloids, micelles, and emulsions.
Weedy plants are plants that compete with cultivated plants, those of particular importance include, but are not limited to important invasive and noxious weeds and herbicide resistant biotypes in crop production, such as, Amaranthus species—A. albus, A. blitoides, A. hybridus, A. palmeri, A. powellii, A. retroflexus, A. spinosus, A. tuberculatus, and A. viridis; Ambrosia species—A. trifida, A. artemisifolia; Lolium species—L. multiflorum, L. rigidum, L perenne; Digitaria species—D. insularis; Euphorbia species—E. heterophylla; Kochia species—K. scoparia; Sorghum species—S. halepense; Conyza species—C. bonariensis, C. canadensis, C. sumatrensis; Chloris species—C. truncate; Echinochola species—E. colona, E. crus-galli; Eleusine species—E. indica; Poa species—P. annua; Plantago species—P. lanceolate, Avena species—A. fatua; Chenopodium species—C. album; Setaria species—S. viridis, Abutilon theophrasti, Ipomoea species, Sesbania species, Cassia species, Sida species, Brachiaria, species and Solanum species.
Additional weedy plant species found in cultivated areas include Alopecurus myosuroides, Avena sterilis, Avena sterilis ludoviciana, Brachiaria plantaginea, Bromus diandrus, Bromus rigidus, Cynosurus echinatus, Digitaria ciliaris, Digitaria ischaemum, Digitaria sanguinalis, Echinochloa oryzicola, Echinochloa phyllopogon, Eriochloa punctata, Hordeum glaucum, Hordeum leporinum, Ischaemum rugosum, Leptochloa chinensis, Lolium persicum, Phalaris minor, Phalaris paradoxa, Rottboellia exalta, Setaria faberi, Setaria viridis var, robusta-alba schreiber, Setaria viridis var, robusta-purpurea, Snowdenia polystachea, Sorghum sudanese, Alisma plantago-aquatica, Amaranthus lividus, Amaranthus quitensis, Ammania auriculata, Ammania coccinea, Anthemis cotula, Apera spica-venti, Bacopa rotundifolia, Bidens pilosa, Bidens subalternans, Brassica tournefortii, Bromus tectorum, Camelina microcarpa, Chrysanthemum coronarium, Cuscuta campestris, Cyperus difformis, Damasonium minus, Descurainia sophia, Diplotaxis tenuifolia, Echium plantagineum, Elatine triandra var, pedicellata, Euphorbia heterophylla, Fallopia convolvulus, Fimbristylis miliacea, Galeopsis tetrahit, Galium spurium, Helianthus annuus, Iva xanthifolia, Ixophorus unisetus, Ipomoea indica, Ipomoea purpurea, Ipomoea sepiaria, Ipomoea aquatic, Ipomoea triloba, Lactuca serriola, Limnocharis flava, Limnophila erecta, Limnophila sessiliflora, Lindernia dubia, Lindernia dubia var, major, Lindernia micrantha, Lindernia procumbens, Mesembryanthemum crystallinum, Monochoria korsakowii, Monochoria vaginalis, Neslia paniculata, Papaver rhoeas, Parthenium hysterophorus, Pentzia suffruticosa, Phalaris minor, Raphanus raphanistrum, Raphanus sativus, Rapistrum rugosum, Rotala indica var, uliginosa, Sagittaria guyanensis, Sagittaria montevidensis, Sagittaria pygmaea, Salsola iberica, Scirpus juncoides var, ohwianus, Scirpus mucronatus, Setaria lutescens, Sida spinosa, Sinapis arvensis, Sisymbrium orientale, Sisymbrium thellungii, Solanum ptycanthum, Sonchus aspen, Sonchus oleraceus, Sorghum bicolor, Stellaria media, Thlaspi arvense, Xanthium strumarium, Arctotheca calendula, Conyza sumatrensis, Crassocephalum crepidiodes, Cuphea carthagenenis, Epilobium adenocaulon, Erigeron philadelphicus, Landoltia punctata, Lepidium virginicum, Monochoria korsakowii, Solanum americanum, Solanum nigrum, Vulpia bromoides, Youngia japonica, Hydrilla verticillata, Carduus nutans, Carduus pycnocephalus, Centaurea solstitialis, Cirsium arvense, Commelina diffusa, Convolvulus arvensis, Daucus carota, Digitaria ischaemum, Echinochloa crus-pavonis, Fimbristylis miliacea, Galeopsis tetrahit, Galium spurium, Limnophila erecta, Matricaria perforate, Papaver rhoeas, Ranunculus acris, Soliva sessilis, Sphenoclea zeylanica, Stellaria media, Nassella trichotoma, Stipa neesiana, Agrostis stolonifera, Polygonum aviculare, Alopecurus japonicus, Beckmannia syzigachne, Bromus tectorum, Chloris inflate, Echinochloa erecta, Portulaca oleracea, and Senecio vulgaris. It is believed that all plants contain a glutamine synthetase (GS) gene in their genome, the sequence of which can be isolated and polynucleotides made according to the methods of the present invention that are useful for regulation, suppressing or delaying the expression of the target GS gene in the plants and the growth or development of the treated plants.
Some cultivated plants may also be weedy plants when they occur in unwanted environments. For example, corn plants growing in a soybean field. Transgenic crops with one or more herbicide tolerances will need specialized methods of management to control weeds and volunteer crop plants. The present invention enables the targeting of a transgene for herbicide tolerance to permit the treated plants to become sensitive to the herbicide. For example, transgene GS DNA sequences in transgenic events that include but are not limited to DP-004114-3, DAS-44406-6, DAS-68416-4, T304-40XGHB119, LLRICE601, TC-6275, LLCotton25, MS1 & RF1/RF2, Topas 19/2, Line 1507, MS6, GU262, A5547-127, T-120-7, W62, W98, A2704-12, A2704-21, A5547-35, and B16.
A “trigger” or “trigger polynucleotide” of the present invention is a polynucleotide molecule that is homologous or complementary to a target gene polynucleotide. The trigger polynucleotide molecules modulate expression of the target gene when topically applied to a plant surface with a transfer agent, whereby a plant treated with said composition has its growth or development or reproductive ability regulated, suppressed or delayed or said plant is more sensitive to a GS inhibitor herbicide as a result of said polynucleotide containing composition relative to a plant not treated with a composition containing the trigger molecule. Trigger polynucleotides disclosed herein are generally described in relation to the target gene sequence and maybe used in the sense (homologous) or antisense (complementary) orientation as single stranded molecules or comprise both strands as double stranded molecules or nucleotide variants and modified nucleotides thereof depending on the various regions of a gene being targeted.
It is contemplated that the composition of the present invention will contain multiple polynucleotides and herbicides that include but not limited to GS gene trigger polynucleotides and a GS inhibitor herbicide and anyone or more additional herbicide target gene trigger polynucleotides and the related herbicides and anyone or more additional essential gene trigger polynucleotides. Essential genes are genes in a plant that provide key enzymes or other proteins, for example, a biosynthetic enzyme, metabolizing enzyme, receptor, signal transduction protein, structural gene product, transcription factor, or transport protein; or regulating RNAs, such as, microRNAs, that are essential to the growth or survival of the organism or cell or involved in the normal growth and development of the plant (Meinke, et al., Trends Plant Sci. 2008 September; 13(9):483-91). The suppression of an essential gene enhances the effect of a herbicide that affects the function of a gene product different than the suppressed essential gene. The compositions of the present invention can include various trigger polynucleotides that modulate the expression of an essential gene other than a GS gene.
Herbicides for which transgenes for plant tolerance have been demonstrated and the method of the present invention can be applied, include but are not limited to: auxin-like herbicides, glyphosate, glufosinate, sulfonylureas, imidazolinones, bromoxynil, delapon, dicamba, cyclohezanedione, protoporphyrionogen oxidase inhibitors, 4-hydroxyphenyl-pyruvate-dioxygenase inhibitors herbicides. For example, transgenes and their polynucleotide molecules that encode proteins involved in herbicide tolerance are known in the art, and include, but are not limited to an 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), for example, as more fully described in U.S. Pat. No. 7,807,791 (SEQ ID NO: 5); U.S. Pat. Nos. 6,248,876 B1; 5,627,061; 5,804,425; 5,633,435; 5,145,783; 4,971,908; 5,3372,910; 5,188,642; 4,940,835; 5,866,775; 6,225,114 B1; 6,130,366; 5,3370,667; 4,535,060; 4,769,061; 5,633,448; 5,510,471; U.S. Pat. No. Re. 36,449; U.S. Pat. Nos. RE 37,287 E; and 5,491,288; tolerance to sulfonylurea and/or imidazolinone, for example, as described more fully in U.S. Pat. Nos. 5,605,011; 5,013,659; 5,141,870; 5,767,361; 5,7337,180; 5,304,732; 4,761,373; 5,3337,107; 5,928,937; and 5,378,824; and international publication WO 96/33270; tolerance to hydroxyphenylpyruvatedioxygenases inhibiting herbicides in plants are described in U.S. Pat. Nos. 6,245,968 B1; 6,268,549; and 6,069,115; US Pat. Pub. 20110191897 and U.S. Pat. No. 7,3372,379 SEQ ID NO: 3; U.S. Pat. Nos. 7,935,869; 7,304,209, SEQ ID NOs: 1, 3, 5 and 15; aryloxyalkanoate dioxygenase polynucleotides, which confer tolerance to 2,4-D and other phenoxy auxin herbicides as well as to aryloxyphenoxypropionate herbicides as described, for example, in WO2005/107437; U.S. Pat. No. 7,838,733 SEQ ID NO: 5;) and dicamba-tolerance polynucleotides as described, for example, in Herman et al. (2005) J. Biol. Chem. 280: 24759-24767. Other examples of herbicide-tolerance traits include those conferred by polynucleotides encoding an exogenous phosphinothricin acetyltransferase, as described in U.S. Pat. Nos. 5,969,213; 5,489,520; 5,550,3378; 5,874,265; 5,919,675; 5,561,236; 5,648,477; 5,646,024; 6,177,616; and 5,879,903. Plants containing an exogenous phosphinothricin acetyltransferase can exhibit improved tolerance to glufosinate herbicides, which inhibit the enzyme glutamine synthetase. Additionally, herbicide-tolerance polynucleotides include those conferred by polynucleotides conferring altered protoporphyrinogen oxidase (protox) activity, as described in U.S. Pat. Nos. 6,288,306 B1; 6,282,837 B1; and 5,767,373; and WO 01/12825. Plants containing such polynucleotides can exhibit improved tolerance to any of a variety of herbicides which target the protox enzyme (also referred to as protox inhibitors). Polynucleotides encoding a glyphosate oxidoreductase and a glyphosate-N-acetyl transferase (GOX described in U.S. Pat. No. 5,463,175 and GAT described in U.S. Patent publication 20030083480, dicamba monooxygenase U.S. Patent publication 20030135879, all of which are incorporated herein by reference); a polynucleotide molecule encoding bromoxynil nitrilase (Bxn described in U.S. Pat. No. 4,810,648 for Bromoxynil tolerance, which is incorporated herein by reference); a polynucleotide molecule encoding phytoene desaturase (crtI) described in Misawa et al, (1993) Plant J. 4:833-840 and Misawa et al, (1994) Plant J. 6:481-489 for norflurazon tolerance; a polynucleotide molecule encoding acetohydroxyacid synthase (AHAS, aka ALS) described in Sathasiivan et al. (1990) Nucl. Acids Res. 18:3378-2193 for tolerance to sulfonylurea herbicides; and the bar gene described in DeBlock, et al. (1987) EMBO J. 6:2513-2519 for glufosinate and bialaphos tolerance. The transgenic coding regions and regulatory elements of the herbicide tolerance genes are targets in which polynucleotide triggers and herbicides can be included in the composition of the present invention.
The compositions include a component that is a GS inhibitor herbicide, which include members of the Phosphinic acids herbicide group such as glufosinate-ammonium and bialaphos.
Numerous herbicides with similar or different modes of action (herein referred to as co-herbicides) are available that can be added to the composition of the present invention, for example, members of the herbicide families that include but are not limited to amide herbicides, aromatic acid herbicides, arsenical herbicides, benzothiazole herbicides, benzoylcyclohexanedione herbicides, benzofuranyl alkylsulfonate herbicides, carbamate herbicides, cyclohexene oxime herbicides, cyclopropylisoxazole herbicides, dicarboximide herbicides, dinitroaniline herbicides, dinitrophenol herbicides, diphenyl ether herbicides, dithiocarbamate herbicides, halogenated aliphatic herbicides, imidazolinone herbicides, inorganic herbicides, nitrile herbicides, organophosphorus herbicides, oxadiazolone herbicides, oxazole herbicides, phenoxy herbicides, phenylenediamine herbicides, pyrazole herbicides, pyridazine herbicides, pyridazinone herbicides, pyridine herbicides, pyrimidinediamine herbicides, pyrimidinyloxybenzylamine herbicides, quaternary ammonium herbicides, thiocarbamate herbicides, thiocarbonate herbicides, thiourea herbicides, triazine herbicides, triazinone herbicides, triazole herbicides, triazolone herbicides, triazolopyrimidine herbicides, uracil herbicides, and urea herbicides. In particular, the rates of use of the added herbicides can be reduced in compositions comprising the polynucleotides of the invention. Use rate reductions of the additional added herbicides can be 10-25 percent, 26-50 percent, 51-75 percent or more can be achieved that enhance the activity of the polynucleotides and herbicide composition and is contemplated as an aspect of the invention. Representative co-herbicides of the families include but are not limited to acetochlor, acifluorfen, acifluorfen-sodium, aclonifen, acrolein, alachlor, alloxydim, allyl alcohol, ametryn, amicarbazone, amidosulfuron, aminopyralid, amitrole, ammonium sulfamate, anilofos, asulam, atraton, atrazine, azimsulfuron, BCPC, beflubutamid, benazolin, benfluralin, benfuresate, bensulfuron, bensulfuron-methyl, bensulide, bentazone, benzfendizone, benzobicyclon, benzofenap, bifenox, bilanafos, bispyribac, bispyribac-sodium, borax, bromacil, bromobutide, bromoxynil, butachlor, butafenacil, butamifos, butralin, butroxydim, butylate, cacodylic acid, calcium chlorate, cafenstrole, carbetamide, carfentrazone, carfentrazone-ethyl, CDEA, CEPC, chlorflurenol, chlorflurenol-methyl, chloridazon, chlorimuron, chlorimuron-ethyl, chloroacetic acid, chlorotoluron, chlorpropham, chlorsulfuron, chlorthal, chlorthal-dimethyl, cinidon-ethyl, cinmethylin, cinosulfuron, cisanilide, clethodim, clodinafop, clodinafop-propargyl, clomazone, clomeprop, clopyralid, cloransulam, cloransulam-methyl, CMA, 4-CPB, CPMF, 4-CPP, CPPC, cresol, cumyluron, cyanamide, cyanazine, cycloate, cyclosulfamuron, cycloxydim, cyhalofop, cyhalofop-butyl, 2,4-D, 3,4-DA, daimuron, dalapon, dazomet, 2,4-DB, 3,4-DB, 2,4-DEB, desmedipham, dicamba, dichlobenil, ortho-dichlorobenzene, para-dichlorobenzene, dichlorprop, dichlorprop-P, diclofop, diclofop-methyl, diclosulam, difenzoquat, difenzoquat metilsulfate, diflufenican, diflufenzopyr, dimefuron, dimepiperate, dimethachlor, dimethametryn, dimethenamid, dimethenamid-P, dimethipin, dimethylarsinic acid, dinitramine, dinoterb, diphenamid, diquat, diquat dibromide, dithiopyr, diuron, DNOC, 3,4-DP, DSMA, EBEP, endothal, EPTC, esprocarb, ethalfluralin, ethametsulfuron, ethametsulfuron-methyl, ethofumesate, ethoxyfen, ethoxysulfuron, etobenzanid, fenoxaprop-P, fenoxaprop-P-ethyl, fentrazamide, ferrous sulfate, flamprop-M, flazasulfuron, florasulam, fluazifop, fluazifop-butyl, fluazifop-P, fluazifop-P-butyl, flucarbazone, flucarbazone-sodium, flucetosulfuron, fluchloralin, flufenacet, flufenpyr, flufenpyr-ethyl, flumetsulam, flumiclorac, flumiclorac-pentyl, flumioxazin, fluometuron, fluoroglycofen, fluoroglycofen-ethyl, flupropanate, flupyrsulfuron, flupyrsulfuron-methyl-sodium, flurenol, fluridone, fluorochloridone, fluoroxypyr, flurtamone, fluthiacet, fluthiacet-methyl, fomesafen, foramsulfuron, fosamine, glufosinate, glufosinate-ammonium, glyphosate, halosulfuron, halosulfuron-methyl, haloxyfop, haloxyfop-P, HC-252, hexazinone, imazamethabenz, imazamethabenz-methyl, imazamox, imazapic, imazapyr, imazaquin, imazethapyr, imazosulfuron, indanofan, iodomethane, iodosulfuron, iodosulfuron-methyl-sodium, ioxynil, isoproturon, isouron, isoxaben, isoxachlortole, isoxaflutole, karbutilate, lactofen, lenacil, linuron, MAA, MAMA, MCPA, MCPA-thioethyl, MCPB, mecoprop, mecoprop-P, mefenacet, mefluidide, mesosulfuron, mesosulfuron-methyl, mesotrione, metam, metamifop, metamitron, metazachlor, methabenzthiazuron, methylarsonic acid, methyldymron, methyl isothiocyanate, metobenzuron, metolachlor, S-metolachlor, metosulam, metoxuron, metribuzin, metsulfuron, metsulfuron-methyl, MK-66, molinate, monolinuron, MSMA, naproanilide, napropamide, naptalam, neburon, nicosulfuron, nonanoic acid, norflurazon, oleic acid (fatty acids), orbencarb, orthosulfamuron, oryzalin, oxadiargyl, oxadiazon, oxasulfuron, oxaziclomefone, oxyfluorfen, paraquat, paraquat dichloride, pebulate, pendimethalin, penoxsulam, pentachlorophenol, pentanochlor, pentoxazone, pethoxamid, petrolium oils, phenmedipham, phenmedipham-ethyl, picloram, picolinafen, pinoxaden, piperophos, potassium arsenite, potassium azide, pretilachlor, primisulfuron, primisulfuron-methyl, prodiamine, profluazol, profoxydim, prometon, prometryn, propachlor, propanil, propaquizafop, propazine, propham, propisochlor, propoxycarbazone, propoxycarbazone-sodium, propyzamide, prosulfocarb, prosulfuron, pyraclonil, pyraflufen, pyraflufen-ethyl, pyrazolynate, pyrazosulfuron, pyrazosulfuron-ethyl, pyrazoxyfen, pyribenzoxim, pyributicarb, pyridafol, pyridate, pyriftalid, pyriminobac, pyriminobac-methyl, pyrimisulfan, pyrithiobac, pyrithiobac-sodium, quinclorac, quinmerac, quinoclamine, quizalofop, quizalofop-P, rimsulfuron, sethoxydim, siduron, simazine, simetryn, SMA, sodium arsenite, sodium azide, sodium chlorate, sulcotrione, sulfentrazone, sulfometuron, sulfometuron-methyl, sulfosate, sulfosulfuron, sulfuric acid, tar oils, 2,3,6-TBA, TCA, TCA-sodium, tebuthiuron, tepraloxydim, terbacil, terbumeton, terbuthylazine, terbutryn, thenylchlor, thiazopyr, thifensulfuron, thifensulfuron-methyl, thiobencarb, tiocarbazil, topramezone, tralkoxydim, tri-allate, triasulfuron, triaziflam, tribenuron, tribenuron-methyl, tricamba, triclopyr, trietazine, trifloxysulfuron, trifloxysulfuron-sodium, trifluralin, triflusulfuron, triflusulfuron-methyl, trihydroxytriazine, tritosulfuron, [3-[2-chloro-4-fluoro-5-(-methyl-6-trifluoromethyl-2,4-dioxo-,2,3,4-t-etrahydropyrimidin-3-yl)phenoxy]-2-pyridyloxy]acetic acid ethyl ester (CAS RN 353292-3-6), 4-[(4,5-dihydro-3-methoxy-4-methyl-5-oxo)-H-,2,4-triazol-1-ylcarbonyl-sulfamoyl]-5-methylthiophene-3-carboxylic acid (BAY636), BAY747 (CAS RN 33504-84-2), topramezone (CAS RN 2063-68-8), 4-hydroxy-3-[[2-[(2-methoxyethoxy)methyl]-6-(trifluoro-methyl)-3-pyridi-nyl]carbonyl]-bicyclo[3.2]oct-3-en-2-one (CAS RN 35200-68-5), and 4-hydroxy-3-[[2-(3-methoxypropyl)-6-(difluoromethyl)-3-pyridinyl]carbonyl]-bicyclo[3.2]oct-3-en-2-one. Additionally, including herbicidal compounds of unspecified modes of action as described in CN101279950A, CN101279951A, DE10000600A1, DE10116399A1, DE102004054666A1, DE102005014638A1, DE102005014906A1, DE102007012168A1, DE102010042866A1, DE10204951A1, DE10234875A1, DE10234876A1, DE10256353A1, DE10256354A1, DE10256367A1, EP1157991A2, EP1238586A1, EP2147919A1, EP2160098A2, JP03968012B2, JP2001253874A, JP2002080454A, JP2002138075A, JP2002145707A, JP2002220389A, JP2003064059A, JP2003096059A, JP2004051628A, JP2004107228A, JP2005008583A, JP2005239675A, JP2005314407A, JP2006232824A, JP2006282552A, JP2007153847A, JP2007161701A, JP2007182404A, JP2008074840A, JP2008074841A, JP2008133207A, JP2008133218A, JP2008169121A, JP2009067739A, JP2009114128A, JP2009126792A, JP2009137851A, US20060111241A1, US20090036311A1, US20090054240A1, US20090215628A1, US20100099561A1, US20100152443A1, US20110105329A1, US20110201501A1, WO2001055066A2, WO2001056975A1, WO2001056979A1, WO2001090071A2, WO2001090080A1, WO2002002540A1, WO2002028182A1, WO2002040473A1, WO2002044173A2, WO2003000679A2, WO2003006422A1, WO2003013247A1, WO2003016308A1, WO2003020704A1, WO2003022051A1, WO2003022831A1, WO2003022843A1, WO2003029243A2, WO2003037085A1, WO2003037878A1, WO2003045878A2, WO2003050087A2, WO2003051823A1, WO2003051824A1, WO2003051846A2, WO2003076409A1, WO2003087067A1, WO2003090539A1, WO2003091217A1, WO2003093269A2, WO2003104206A2, WO2004002947A1, WO2004002981A2, WO2004011429A1, WO2004029060A1, WO2004035545A2, WO2004035563A1, WO2004035564A1, WO2004037787A1, WO2004067518A1, WO2004067527A1, WO2004077950A1, WO2005000824A1, WO2005007627A1, WO2005040152A1, WO2005047233A1, WO2005047281A1, WO2005061443A2, WO2005061464A1, WO2005068434A1, WO2005070889A1, WO2005089551A1, WO2005095335A1, WO2006006569A1, WO2006024820A1, WO2006029828A1, WO2006029829A1, WO2006037945A1, WO2006050803A1, WO2006090792A1, WO2006123088A2, WO2006125687A1, WO2006125688A1, WO2007003294A1, WO2007026834A1, WO2007071900A1, WO2007077201A1, WO2007077247A1, WO2007096576A1, WO2007119434A1, WO2007134984A1, WO2008009908A1, WO2008029084A1, WO2008059948A1, WO2008071918A1, WO2008074991A1, WO2008084073A1, WO2008100426A2, WO2008102908A1, WO2008152072A2, WO2008152073A2, WO2009000757A1, WO2009005297A2, WO2009035150A2, WO2009063180A1, WO2009068170A2, WO2009068171A2, WO2009086041A1, WO2009090401A2, WO2009090402A2, WO2009115788A1, WO2009116558A1, WO2009152995A1, WO2009158258A1, WO2010012649A1, WO2010012649A1, WO2010026989A1, WO2010034153A1, WO2010049270A1, WO2010049369A1, WO2010049405A1, WO2010049414A1, WO2010063422A1, WO2010069802A1, WO2010078906A2, WO2010078912A1, WO2010104217A1, WO2010108611A1, WO2010112826A3, WO2010116122A3, WO2010119906A1, WO2010130970A1, WO2011003776A2, WO2011035874A1, WO2011065451A1, all of which are incorporated herein by reference.
An agronomic field in need of plant control is treated by application of the composition directly to the surface of the growing plants, such as by a spray. For example, the method is applied to control weeds in a field of crop plants by spraying the field with the composition. The composition can be provided as a tank mix, a sequential treatment of components (generally the polynucleotide containing composition followed by the herbicide), or a simultaneous treatment or mixing of one or more of the components of the composition from separate containers. Treatment of the field can occur as often as needed to provide weed control and the components of the composition can be adjusted to target specific weed species or weed families through utilization of specific polynucleotides or polynucleotide compositions capable of selectively targeting the specific species or plant family to be controlled. The composition can be applied at effective use rates according to the time of application to the field, for example, preplant, at planting, post planting, post harvest. GS inhibitor herbicides can be applied to a field at rates of 100 to 500 g ai/ha (active ingredient per hectare) or more. The polynucleotides of the composition can be applied at rates of 1 to 30 grams per acre depending on the number of trigger molecules needed for the scope of weeds in the field.
Crop plants in which weed control is needed include but are not limited to, i) corn, soybean, cotton, canola, sugar beet, alfalfa, sugarcane, rice, and wheat; ii) vegetable plants including, but not limited to, tomato, sweet pepper, hot pepper, melon, watermelon, cucumber, eggplant, cauliflower, broccoli, lettuce, spinach, onion, peas, carrots, sweet corn, Chinese cabbage, leek, fennel, pumpkin, squash or gourd, radish, Brussels sprouts, tomatillo, garden beans, dry beans, or okra; iii) culinary plants including, but not limited to, basil, parsley, coffee, or tea; or, iv) fruit plants including but not limited to apple, pear, cherry, peach, plum, apricot, banana, plantain, table grape, wine grape, citrus, avocado, mango, or berry; v) a tree grown for ornamental or commercial use, including, but not limited to, a fruit or nut tree; or, vi) an ornamental plant (e.g., an ornamental flowering plant or shrub or turf grass). The methods and compositions provided herein can also be applied to plants produced by a cutting, cloning, or grafting process (i.e., a plant not grown from a seed) include fruit trees and plants that include, but are not limited to, citrus, apples, avocados, tomatoes, eggplant, cucumber, melons, watermelons, and grapes as well as various ornamental plants.
Pesticidal Mixtures
The polynucleotide compositions may also be used as mixtures with various agricultural chemicals and/or insecticides, miticides and fungicides, pesticidal and biopesticidal agents. Examples include but are not limited to azinphos-methyl, acephate, isoxathion, isofenphos, ethion, etrimfos, oxydemeton-methyl, oxydeprofos, quinalphos, chlorpyrifos, chlorpyrifos-methyl, chlorfenvinphos, cyanophos, dioxabenzofos, dichlorvos, disulfoton, dimethylvinphos, dimethoate, sulprofos, diazinon, thiometon, tetrachlorvinphos, temephos, tebupirimfos, terbufos, naled, vamidothion, pyraclofos, pyridafenthion, pirimiphos-methyl, fenitrothion, fenthion, phenthoate, flupyrazophos, prothiofos, propaphos, profenofos, phoxime, phosalone, phosmet, formothion, phorate, malathion, mecarb am, mesulfenfos, methamidophos, methidathion, parathion, methyl parathion, monocrotophos, trichlorphon, EPN, isazophos, isamidofos, cadusafos, diamidaphos, dichlofenthion, thionazin, fenamiphos, fosthiazate, fosthietan, phosphocarb, DSP, ethoprophos, alanycarb, aldicarb, isoprocarb, ethiofencarb, carbaryl, carbosulfan, xylylcarb, thiodicarb, pirimicarb, fenobucarb, furathiocarb, propoxur, bendiocarb, benfuracarb, methomyl, metolcarb, XMC, carbofuran, aldoxycarb, oxamyl, acrinathrin, allethrin, esfenvalerate, empenthrin, cycloprothrin, cyhalothrin, gamma-cyhalothrin, lambda-cyhalothrin, cyfluthrin, beta-cyfluthrin, cypermethrin, alpha-cypermethrin, zeta-cypermethrin, silafluofen, tetramethrin, tefluthrin, deltamethrin, tralomethrin, bifenthrin, phenothrin, fenvalerate, fenpropathrin, furamethrin, prallethrin, flucythrinate, fluvalinate, flubrocythrinate, permethrin, resmethrin, ethofenprox, cartap, thiocyclam, bensultap, acetamiprid, imidacloprid, clothianidin, dinotefuran, thiacloprid, thiamethoxam, nitenpyram, chlorfluazuron, diflubenzuron, teflubenzuron, triflumuron, novaluron, noviflumuron, bistrifluoron, fluazuron, flucycloxuron, flufenoxuron, hexaflumuron, lufenuron, chromafenozide, tebufenozide, halofenozide, methoxyfenozide, diofenolan, cyromazine, pyriproxyfen, buprofezin, methoprene, hydroprene, kinoprene, triazamate, endosulfan, chlorfenson, chlorobenzilate, dicofol, bromopropylate, acetoprole, fipronil, ethiprole, pyrethrin, rotenone, nicotine sulphate, BT (Bacillus Thuringiensis) agent, spinosad, abamectin, acequinocyl, amidoflumet, amitraz, etoxazole, chinomethionat, clofentezine, fenbutatin oxide, dienochlor, cyhexatin, spirodiclofen, spiromesifen, tetradifon, tebufenpyrad, binapacryl, bifenazate, pyridaben, pyrimidifen, fenazaquin, fenothiocarb, fenpyroximate, fluacrypyrim, fluazinam, flufenzin, hexythiazox, propargite, benzomate, polynactin complex, milbemectin, lufenuron, mecarbam, methiocarb, mevinphos, halfenprox, azadirachtin, diafenthiuron, indoxacarb, emamectin benzoate, potassium oleate, sodium oleate, chlorfenapyr, tolfenpyrad, pymetrozine, fenoxycarb, hydramethylnon, hydroxy propyl starch, pyridalyl, flufenerim, flubendiamide, flonicamid, metaflumizole, lepimectin, TPIC, albendazole, oxibendazole, oxfendazole, trichlamide, fensulfothion, fenbendazole, levamisole hydrochloride, morantel tartrate, dazomet, metam-sodium, triadimefon, hexaconazole, propiconazole, ipconazole, prochloraz, triflumizole, tebuconazole, epoxiconazole, difenoconazole, flusilazole, triadimenol, cyproconazole, metconazole, fluquinconazole, bitertanol, tetraconazole, triticonazole, flutriafol, penconazole, diniconazole, fenbuconazole, bromuconazole, imibenconazole, simeconazole, myclobutanil, hymexazole, imazalil, furametpyr, thifluzamide, etridiazole, oxpoconazole, oxpoconazole fumarate, pefurazoate, prothioconazole, pyrifenox, fenarimol, nuarimol, bupirimate, mepanipyrim, cyprodinil, pyrimethanil, metalaxyl, mefenoxam, oxadixyl, benalaxyl, thiophanate, thiophanate-methyl, benomyl, carbendazim, fuberidazole, thiabendazole, manzeb, propineb, zineb, metiram, maneb, ziram, thiuram, chlorothalonil, ethaboxam, oxycarboxin, carboxin, flutolanil, silthiofam, mepronil, dimethomorph, fenpropidin, fenpropimorph, spiroxamine, tridemorph, dodemorph, flumorph, azoxystrobin, kresoxim-methyl, metominostrobin, orysastrobin, fluoxastrobin, trifloxystrobin, dimoxystrobin, pyraclostrobin, picoxystrobin, iprodione, procymidone, vinclozolin, chlozolinate, flusulfamide, dazomet, methyl isothiocyanate, chloropicrin, methasulfocarb, hydroxyisoxazole, potassium hydroxyisoxazole, echlomezol, D-D, carbam, basic copper chloride, basic copper sulfate, copper nonylphenolsulfonate, oxine copper, DBEDC, anhydrous copper sulfate, copper sulfate pentahydrate, cupric hydroxide, inorganic sulfur, wettable sulfur, lime sulfur, zinc sulfate, fentin, sodium hydrogen carbonate, potassium hydrogen carbonate, sodium hypochlorite, silver, edifenphos, tolclofos-methyl, fosetyl, iprobenfos, dinocap, pyrazophos, carpropamid, fthalide, tricyclazole, pyroquilon, diclocymet, fenoxanil, kasugamycin, validamycin, polyoxins, blasticiden S, oxytetracycline, mildiomycin, streptomycin, rape seed oil, machine oil, benthiavalicarbisopropyl, iprovalicarb, propamocarb, diethofencarb, fluoroimide, fludioxanil, fenpiclonil, quinoxyfen, oxolinic acid, chlorothalonil, captan, folpet, probenazole, acibenzolar-S-methyl, tiadinil, cyflufenamid, fenhexamid, diflumetorim, metrafenone, picobenzamide, proquinazid, famoxadone, cyazofamid, fenamidone, zoxamide, boscalid, cymoxanil, dithianon, fluazinam, dichlofluanide, triforine, isoprothiolane, ferimzone, diclomezine, tecloftalam, pencycuron, chinomethionat, iminoctadine acetate, iminoctadine albesilate, ambam, polycarbamate, thiadiazine, chloroneb, nickel dimethyldithiocarbamate, guazatine, dodecylguanidine-acetate, quintozene, tolylfluanid, anilazine, nitrothalisopropyl, fenitropan, dimethirimol, benthiazole, harpin protein, flumetover, mandipropamide and penthiopyrad.
Polynucleotides
As used herein, the term “DNA”, “DNA molecule”, “DNA polynucleotide molecule” refers to a single-stranded DNA (ssDNA) or double-stranded DNA (dsDNA) molecule of genomic or synthetic origin, such as, a polymer of deoxyribonucleotide bases or a DNA polynucleotide molecule. As used herein, the term “DNA sequence”, “DNA nucleotide sequence” or “DNA polynucleotide sequence” refers to the nucleotide sequence of a DNA molecule. As used herein, the term “RNA”, “RNA molecule”, “RNA polynucleotide molecule” refers to a single-stranded RNA (ssRNA) or double-stranded RNA (dsRNA) molecule of genomic or synthetic origin, such as, a polymer of ribonucleotide bases that comprise single or double stranded regions. Unless otherwise stated, nucleotide sequences in the text of this specification are given, when read from left to right, in the 5′ to 3′ direction. The nomenclature used herein is that required by Title 37 of the United States Code of Federal Regulations § 1.822 and set forth in the tables in WIPO Standard ST.25 (1998), Appendix 2, Tables 1 and 3.
As used herein, “polynucleotide” refers to a DNA or RNA molecule containing multiple nucleotides and generally refers both to “oligonucleotides” (a polynucleotide molecule of typically 50 or fewer nucleotides in length) and polynucleotides of 51 or more nucleotides. Embodiments of this invention include compositions including oligonucleotides having a length of 18-25 nucleotides (18-mers, 19-mers, 20-mers, 21-mers, 22-mers, 23-mers, 24-mers, or 25-mers) for example, oligonucleotides SEQ ID NOs:1444-2045 or fragments thereof, or medium-length polynucleotides having a length of 26 or more nucleotides (polynucleotides of 26, 27, 28, 29, 30, 337, 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, about 65, about 70, about 75, about 80, about 85, about 90, about 95, about 100, about 110, about 120, about 130, about 140, about 150, about 160, about 170, about 180, about 190, about 200, about 210, about 220, about 230, about 240, about 250, about 260, about 270, about 280, about 290, or about 300 nucleotides), for example, oligonucleotides of SEQ ID NOs: 60-1443 or fragments thereof or long polynucleotides having a length greater than about 300 nucleotides (for example, polynucleotides of between about 300 to about 400 nucleotides, between about 400 to about 500 nucleotides, between about 500 to about 600 nucleotides, between about 600 to about 700 nucleotides, between about 700 to about 800 nucleotides, between about 800 to about 900 nucleotides, between about 900 to about 1000 nucleotides, between about 300 to about 500 nucleotides, between about 300 to about 600 nucleotides, between about 300 to about 700 nucleotides, between about 300 to about 800 nucleotides, between about 300 to about 900 nucleotides, or about 1000 nucleotides in length, or even greater than about 1000 nucleotides in length, for example up to the entire length of a target gene including coding or non-coding or both coding and non-coding portions of the target gene), for example, polynucleotides of Table 1 (SEQ ID NOs: 1-59), wherein the selected polynucleotides or fragments thereof are homologous or complementary to SEQ ID NOs: 1-59, suppresses, represses or otherwise delays the expression of the target GS gene. A target gene comprises any polynucleotide molecule in a plant cell or fragment thereof for which the modulation of the expression of the target gene is provided by the methods and compositions of the present invention. Where a polynucleotide is double-stranded, its length can be similarly described in terms of base pairs. Oligonucleotides and polynucleotides of the present invention can be made that are essentially identical or essentially complementary to adjacent genetic elements of a gene, for example, spanning the junction region of an intron and exon, the junction region of a promoter and a transcribed region, the junction region of a 5′ leader and a coding sequence, the junction of a 3′ untranslated region and a coding sequence.
Polynucleotide compositions used in the various embodiments of this invention include compositions including oligonucleotides or polynucleotides or a mixture of both, including RNA or DNA or RNA/DNA hybrids or chemically modified oligonucleotides or polynucleotides or a mixture thereof. In some embodiments, the polynucleotide may be a combination of ribonucleotides and deoxyribonucleotides, for example, synthetic polynucleotides consisting mainly of ribonucleotides but with one or more terminal deoxyribonucleotides or synthetic polynucleotides consisting mainly of deoxyribonucleotides but with one or more terminal dideoxyribonucleotides. In some embodiments, the polynucleotide includes non-canonical nucleotides such as inosine, thiouridine, or pseudouridine. In some embodiments, the polynucleotide includes chemically modified nucleotides. Examples of chemically modified oligonucleotides or polynucleotides are well known in the art; see, for example, US Patent Publication 20110171287, US Patent Publication 20110171176, and US Patent Publication 20110152353, US Patent Publication, 20110152346, US Patent Publication 20110160082, herein incorporated by reference. For example, including but not limited to the naturally occurring phosphodiester backbone of an oligonucleotide or polynucleotide can be partially or completely modified with phosphorothioate, phosphorodithioate, or methylphosphonate internucleotide linkage modifications, modified nucleoside bases or modified sugars can be used in oligonucleotide or polynucleotide synthesis, and oligonucleotides or polynucleotides can be labeled with a fluorescent moiety (for example, fluorescein or rhodamine) or other label (for example, biotin).
The polynucleotides can be single- or double-stranded RNA or single- or double-stranded DNA or double-stranded DNA/RNA hybrids or modified analogues thereof, and can be of oligonucleotide lengths or longer. In more specific embodiments of the invention the polynucleotides that provide single-stranded RNA in the plant cell are selected from the group consisting of (a) a single-stranded RNA molecule (ssRNA), (b) a single-stranded RNA molecule that self-hybridizes to form a double-stranded RNA molecule, (c) a double-stranded RNA molecule (dsRNA), (d) a single-stranded DNA molecule (ssDNA), (e) a single-stranded DNA molecule that self-hybridizes to form a double-stranded DNA molecule, and (f) a single-stranded DNA molecule including a modified Pol III gene that is transcribed to an RNA molecule, (g) a double-stranded DNA molecule (dsDNA), (h) a double-stranded DNA molecule including a modified Pol III gene that is transcribed to an RNA molecule, (i) a double-stranded, hybridized RNA/DNA molecule, or combinations thereof. In some embodiments these polynucleotides include chemically modified nucleotides or non-canonical nucleotides. In some embodiments, the oligonucleotides may be blunt-ended or may comprise a 3′ overhang of from 1-5 nucleotides of at least one or both of the strands. Other configurations of the oligonucleotide are known in the field and are contemplated herein. In embodiments of the method the polynucleotides include double-stranded DNA formed by intramolecular hybridization, double-stranded DNA formed by intermolecular hybridization, double-stranded RNA formed by intramolecular hybridization, or double-stranded RNA formed by intermolecular hybridization. In one embodiment the polynucleotides include single-stranded DNA or single-stranded RNA that self-hybridizes to form a hairpin structure having an at least partially double-stranded structure including at least one segment that will hybridize to RNA transcribed from the gene targeted for suppression. Not intending to be bound by any mechanism, it is believed that such polynucleotides are or will produce single-stranded RNA with at least one segment that will hybridize to RNA transcribed from the gene targeted for suppression. In certain other embodiments the polynucleotides further includes a promoter, generally a promoter functional in a plant, for example, a pol II promoter, a pol III promoter, a pol IV promoter, or a pol V promoter.
The term “gene” refers to chromosomal DNA, plasmid DNA, cDNA, intron and exon DNA, artificial DNA polynucleotide, or other DNA that encodes a peptide, polypeptide, protein, or RNA transcript molecule, and the genetic elements flanking the coding sequence that are involved in the regulation of expression, such as, promoter regions, 5′ leader regions, 3′ untranslated regions. Any of the components of the gene are potential targets for the oligonucleotides and polynucleotides of the present invention.
The polynucleotide molecules of the present invention are designed to modulate expression by inducing regulation or suppression of an endogenous GS gene in a plant and are designed to have a nucleotide sequence essentially identical or essentially complementary to the nucleotide sequence of an endogenous GS gene of a plant or to the sequence of RNA transcribed from an endogenous GS gene of a plant, including a transgene in a plant that provides for a herbicide resistant GS enzyme, which can be coding sequence or non-coding sequence. Effective molecules that modulate expression are referred to as “a trigger molecule, or trigger polynucleotide”. By “essentially identical” or “essentially complementary” is meant that the trigger polynucleotides (or at least one strand of a double-stranded polynucleotide or portion thereof, or a portion of a single strand polynucleotide) are designed to hybridize to the endogenous gene noncoding sequence or to RNA transcribed (known as messenger RNA or an RNA transcript) from the endogenous gene to effect regulation or suppression of expression of the endogenous gene. Trigger molecules are identified by “tiling” the gene targets with partially overlapping probes or non-overlapping probes of antisense or sense polynucleotides that are essentially identical or essentially complementary to the nucleotide sequence of an endogenous gene. Multiple target sequences can be aligned and sequence regions with homology in common, according to the methods of the present invention, are identified as potential trigger molecules for the multiple targets. Multiple trigger molecules of various lengths, for example 18-25 nucleotides, 26-50 nucleotides, 51-100 nucleotides, 101-200 nucleotides, 201-300 nucleotides or more can be pooled into a few treatments in order to investigate polynucleotide molecules that cover a portion of a gene sequence (for example, a portion of a coding versus a portion of a noncoding region, or a 5′ versus a 3′ portion of a gene) or an entire gene sequence including coding and noncoding regions of a target gene. Polynucleotide molecules of the pooled trigger molecules can be divided into smaller pools or single molecules in order to identify trigger molecules that provide the desired effect.
The target gene RNA and DNA polynucleotide molecules (Table 1, SEQ ID NOs: 1-59) are sequenced by any number of available methods and equipment. Some of the sequencing technologies are available commercially, such as the sequencing-by-hybridization platform from Affymetrix Inc. (Sunnyvale, Calif.) and the sequencing-by-synthesis platforms from 454 Life Sciences (Bradford, Conn.), Illumina/Solexa (Hayward, Calif.) and Helicos Biosciences (Cambridge, Mass.), and the sequencing-by-ligation platform from Applied Biosystems (Foster City, Calif.), as described below. In addition to the single molecule sequencing performed using sequencing-by-synthesis of Helicos Biosciences, other single molecule sequencing technologies are encompassed by the method of the invention and include the SMRT™ technology of Pacific Biosciences, the Ion Torrent™ technology, and nanopore sequencing being developed for example, by Oxford Nanopore Technologies. A GS target gene comprising DNA or RNA can be isolated using primers or probes essentially complementary or essentially homologous to SEQ ID NOs: 1-59 or a fragment thereof. A polymerase chain reaction (PCR) gene fragment can be produced using primers essentially complementary or essentially homologous to SEQ ID NOs: 1-59 or a fragment thereof that is useful to isolate a GS gene from a plant genome. SEQ ID NOs: 1-59 or fragments thereof can be used in various sequence capture technologies to isolate additional target gene sequences, for example, including but not limited to Roche NimbleGen® (Madison, Wis.) and Streptavdin-coupled Dynabeads® (Life Technologies, Grand Island, N.Y.) and US20110015084, herein incorporated by reference in its entirety.
Embodiments of functional single-stranded polynucleotides have sequence complementarity that need not be 100 percent, but is at least sufficient to permit hybridization to RNA transcribed from the target gene or DNA of the target gene to form a duplex to permit a gene silencing mechanism. Thus, in embodiments, a polynucleotide fragment is designed to be essentially identical to, or essentially complementary to, a sequence of 18 or more contiguous nucleotides in either the target GS gene sequence or messenger RNA transcribed from the target gene. By “essentially identical” is meant having 100 percent sequence identity or at least about 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent sequence identity when compared to the sequence of 18 or more contiguous nucleotides in either the target gene or RNA transcribed from the target gene; by “essentially complementary” is meant having 100 percent sequence complementarity or at least about 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent sequence complementarity when compared to the sequence of 18 or more contiguous nucleotides in either the target gene or RNA transcribed from the target gene. In some embodiments of this invention polynucleotide molecules are designed to have 100 percent sequence identity with or complementarity to one allele or one family member of a given target gene (coding or non-coding sequence of a gene for of the present invention); in other embodiments the polynucleotide molecules are designed to have 100 percent sequence identity with or complementarity to multiple alleles or family members of a given target gene.
In certain embodiments, the polynucleotides used in the compositions that are essentially identical or essentially complementary to the target gene or transcript will comprise the predominant nucleic acid in the composition. Thus in certain embodiments, the polynucleotides that are essentially identical or essentially complementary to the target gene or transcript will comprise at least about 50%, 75%, 95%, 98% or 100% of the nucleic acids provided in the composition by either mass or molar concentration. However, in certain embodiments, the polynucleotides that are essentially identical or essentially complementary to the target gene or transcript can comprise at least about 1% to about 50%, about 10% to about 50%, about 20% to about 50%, or about 30% to about 50% of the nucleic acids provided in the composition by either mass or molar concentration. Also provided are compositions where the polynucleotides that are essentially identical or essentially complementary to the target gene or transcript can comprise at least about 1% to 100%, about 10% to 100%, about 20% to about 100%, about 30% to about 50%, or about 50% to a 100% of the nucleic acids provided in the composition by either mass or molar concentration.
“Identity” refers to the degree of similarity between two polynucleic acid or protein sequences. An alignment of the two sequences is performed by a suitable computer program. A widely used and accepted computer program for performing sequence alignments is CLUSTALW v1.6 (Thompson, et al. Nucl. Acids Res., 22: 4673-4680, 1994). The number of matching bases or amino acids is divided by the total number of bases or amino acids, and multiplied by 100 to obtain a percent identity. For example, if two 580 base pair sequences had 145 matched bases, they would be 25 percent identical. If the two compared sequences are of different lengths, the number of matches is divided by the shorter of the two lengths. For example, if there are 100 matched amino acids between a 200 and a 400 amino acid protein, they are 50 percent identical with respect to the shorter sequence. If the shorter sequence is less than 150 bases or 50 amino acids in length, the number of matches are divided by 150 (for nucleic acid bases) or 50 (for amino acids), and multiplied by 100 to obtain a percent identity.
Trigger molecules for specific gene family members can be identified from coding and/or non-coding sequences of gene families of a plant or multiple plants, by aligning and selecting 200-300 polynucleotide fragments from the least homologous regions amongst the aligned sequences and evaluated using topically applied polynucleotides (as sense or anti-sense ssDNA or ssRNA, dsRNA, or dsDNA) to determine their relative effectiveness in inducing the herbicidal phenotype. The effective segments are further subdivided into 50-60 polynucleotide fragments, prioritized by least homology, and reevaluated using topically applied polynucleotides. The effective 50-60 polynucleotide fragments are subdivided into 19-30 polynucleotide fragments, prioritized by least homology, and again evaluated for induction of the yield/quality phenotype. Once relative effectiveness is determined, the fragments are utilized singly, or again evaluated in combination with one or more other fragments to determine the trigger composition or mixture of trigger polynucleotides for providing the yield/quality phenotype.
Trigger molecules for broad activity can be identified from coding and/or non-coding sequences of gene families of a plant or multiple plants, by aligning and selecting 200-300 polynucleotide fragments from the most homologous regions amongst the aligned sequences and evaluated using topically applied polynucleotides (as sense or anti-sense ssDNA or ssRNA, dsRNA, or dsDNA) to determine their relative effectiveness in inducing the yield/quality phenotype. The effective segments are subdivided into 50-60 polynucleotide fragments, prioritized by most homology, and reevaluated using topically applied polynucleotides. The effective 50-60 polynucleotide fragments are subdivided into 19-30 polynucleotide fragments, prioritized by most homology, and again evaluated for induction of the yield/quality phenotype. Once relative effectiveness is determined, the fragments may be utilized singly, or in combination with one or more other fragments to determine the trigger composition or mixture of trigger polynucleotides for providing the yield/quality phenotype.
Methods of making polynucleotides are well known in the art. Chemical synthesis, in vivo synthesis and in vitro synthesis methods and compositions are known in the art and include various viral elements, microbial cells, modified polymerases, and modified nucleotides. Commercial preparation of oligonucleotides often provides two deoxyribonucleotides on the 3′ end of the sense strand. Long polynucleotide molecules can be synthesized from commercially available kits, for example, kits from Applied Biosystems/Ambion (Austin, Tex.) have DNA ligated on the 5′ end in a microbial expression cassette that includes a bacterial T7 polymerase promoter that makes RNA strands that can be assembled into a dsRNA and kits provided by various manufacturers that include T7 RiboMax Express (Promega, Madison, Wis.), AmpliScribe T7-Flash (Epicentre, Madison, Wis.), and TranscriptAid T7 High Yield (Fermentas, Glen Burnie, Md.). dsRNA molecules can be produced from microbial expression cassettes in bacterial cells (Ongvarrasopone et al. ScienceAsia 33:35-39; Yin, Appl. Microbiol. Biotechnol. 84:323-333, 2009; Liu et al., BMC Biotechnology 10:85, 2010) that have regulated or deficient RNase III enzyme activity or the use of various viral vectors to produce sufficient quantities of dsRNA. In the present invention, GS gene fragments are inserted into the microbial expression cassettes in a position in which the fragments are express to produce ssRNA or dsRNA useful in the methods described herein to regulate expression on a target GS gene. Long polynucleotide molecules can also be assembled from multiple RNA or DNA fragments. In some embodiments design parameters such as Reynolds score (Reynolds et al. Nature Biotechnology 22, 326-330 (2004), Tuschl rules (Pei and Tuschl, Nature Methods 3(9): 670-676, 2006), i-score (Nucleic Acids Res 35: e123, 2007), i-Score Designer tool and associated algorithms (Nucleic Acids Res 32: 936-948, 2004. Biochem Biophys Res Commun 316: 1050-1058, 2004, Nucleic Acids Res 32: 893-901, 2004, Cell Cycle 3: 790-5, 2004, Nat. Biotechnol. 23: 995-1001, 2005, Nucleic Acids Res. 35: e27, 2007, BMC Bioinformatics 7: 520, 2006, Nucleic Acids Res. 35: e123, 2007, Nat. Biotechnol. 22: 326-330, 2004) are known in the art and may be used in selecting polynucleotide sequences effective in gene silencing. In some embodiments the sequence of a polynucleotide is screened against the genomic DNA of the intended plant to minimize unintentional silencing of other genes.
The trigger polynucleotide and oligonucleotide molecule compositions of this invention are useful in compositions, such as liquids that comprise the polynucleotide molecules at low concentrations, alone or in combination with other components, for example one or more herbicide molecules, either in the same solution or in separately applied liquids that also provide a transfer agent. While there is no upper limit on the concentrations and dosages of polynucleotide molecules that can useful in the methods, lower effective concentrations and dosages will generally be sought for efficiency. The concentrations can be adjusted in consideration of the volume of spray or treatment applied to plant leaves or other plant part surfaces, such as flower petals, stems, tubers, fruit, anthers, pollen, or seed. In one embodiment, a useful treatment for herbaceous plants using 25-mer oligonucleotide molecules is about 1 nanomole (nmol) of oligonucleotide molecules per plant, for example, from about 0.05 to 1 nmol per plant. Other embodiments for herbaceous plants include useful ranges of about 0.05 to about 100 nmol, or about 0.1 to about 20 nmol, or about 1 nmol to about 10 nmol of polynucleotides per plant. Very large plants, trees, or vines may require correspondingly larger amounts of polynucleotides. When using long dsRNA molecules that can be processed into multiple oligonucleotides, lower concentrations can be used. To illustrate embodiments of the invention, the factor 1×, when applied to oligonucleotide molecules is arbitrarily used to denote a treatment of 0.8 nmol of polynucleotide molecule per plant; 10×, 8 nmol of polynucleotide molecule per plant; and 100×, 80 nmol of polynucleotide molecule per plant.
The polynucleotide compositions of this invention are useful in compositions, such as liquids that comprise polynucleotide molecules, alone or in combination with other components either in the same liquid or in separately applied liquids that provide a transfer agent. As used herein, a transfer agent is an agent that, when combined with a polynucleotide in a composition that is topically applied to a target plant surface, enables the polynucleotide to enter a plant cell. In certain embodiments, a transfer agent is an agent that conditions the surface of plant tissue, e.g., leaves, stems, roots, flowers, or fruits, to permeation by the polynucleotide molecules into plant cells. The transfer of polynucleotides into plant cells can be facilitated by the prior or contemporaneous application of a polynucleotide-transferring agent to the plant tissue. In some embodiments the transferring agent is applied subsequent to the application of the polynucleotide composition. The polynucleotide transfer agent enables a pathway for polynucleotides through cuticle wax barriers, stomata and/or cell wall or membrane barriers into plant cells. Suitable transfer agents to facilitate transfer of the polynucleotide into a plant cell include agents that increase permeability of the exterior of the plant or that increase permeability of plant cells to oligonucleotides or polynucleotides. Such agents to facilitate transfer of the composition into a plant cell include a chemical agent, or a physical agent, or combinations thereof. Chemical agents for conditioning or transfer include (a) surfactants, (b) an organic solvent or an aqueous solution or aqueous mixtures of organic solvents, (c) oxidizing agents, (d) acids, (e) bases, (f) oils, (g) enzymes, or combinations thereof. Embodiments of the method can optionally include an incubation step, a neutralization step (e.g., to neutralize an acid, base, or oxidizing agent, or to inactivate an enzyme), a rinsing step, or combinations thereof. Embodiments of agents or treatments for conditioning of a plant to permeation by polynucleotides include emulsions, reverse emulsions, liposomes, and other micellar-like compositions. Embodiments of agents or treatments for conditioning of a plant to permeation by polynucleotides include counter-ions or other molecules that are known to associate with nucleic acid molecules, e.g., inorganic ammonium ions, alkyl ammonium ions, lithium ions, polyamines such as spermine, spermidine, or putrescine, and other cations. Organic solvents useful in conditioning a plant to permeation by polynucleotides include DMSO, DMF, pyridine, N-pyrrolidine, hexamethylphosphoramide, acetonitrile, dioxane, polypropylene glycol, other solvents miscible with water or that will dissolve phosphonucleotides in non-aqueous systems (such as is used in synthetic reactions). Naturally derived or synthetic oils with or without surfactants or emulsifiers can be used, e.g., plant-sourced oils, crop oils (such as those listed in the 9th Compendium of Herbicide Adjuvants, publicly available on the worldwide web (internet) at herbicide.adjuvants.com can be used, e.g., paraffinic oils, polyol fatty acid esters, or oils with short-chain molecules modified with amides or polyamines such as polyethyleneimine or N-pyrrolidine. Transfer agents include, but are not limited to, organosilicone preparations.
In certain embodiments, an organosilicone preparation that is commercially available as Silwet® L-77 surfactant having CAS Number 27306-78-1 and EPA Number: CAL. REG. NO. 5905-50073-AA, and currently available from Momentive Performance Materials, Albany, N.Y. can be used to prepare a polynucleotide composition. In certain embodiments where a Silwet L-77 organosilicone preparation is used as a pre-spray treatment of plant leaves or other plant surfaces, freshly made concentrations in the range of about 0.015 to about 2 percent by weight (wt percent) (e.g., about 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.5 wt percent) are efficacious in preparing a leaf or other plant surface for transfer of polynucleotide molecules into plant cells from a topical application on the surface. In certain embodiments of the methods and compositions provided herein, a composition that comprises a polynucleotide molecule and an organosilicone preparation comprising Silwet L-77 in the range of about 0.015 to about 2 percent by weight (wt percent) (e.g., about 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.5 wt percent) is used or provided.
In certain embodiments, any of the commercially available organosilicone preparations provided such as the following Breakthru S 321, Breakthru S 200 Cat #67674-67-3, Breakthru OE 441 Cat #68937-55-3, Breakthru S 278 Cat #27306-78-1, Breakthru S 243, Breakthru S 233 Cat #134180-76-0, available from manufacturer Evonik Goldschmidt (Germany), Silwet® HS 429, Silwet® HS 312, Silwet® HS 508, Silwet® HS 604 (Momentive Performance Materials, Albany, N.Y.) can be used as transfer agents in a polynucleotide composition. In certain embodiments where an organosilicone preparation is used as a pre-spray treatment of plant leaves or other surfaces, freshly made concentrations in the range of about 0.015 to about 2 percent by weight (wt percent) (e.g., about 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.5 wt percent) are efficacious in preparing a leaf or other plant surface for transfer of polynucleotide molecules into plant cells from a topical application on the surface. In certain embodiments of the methods and compositions provided herein, a composition that comprises a polynucleotide molecule and an organosilicone preparation in the range of about 0.015 to about 2 percent by weight (wt percent) (e.g., about 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.5 wt percent) is used or provided.
Organosilicone preparations used in the methods and compositions provided herein can comprise one or more effective organosilicone compounds. As used herein, the phrase “effective organosilicone compound” is used to describe any organosilicone compound that is found in an organosilicone preparation that enables a polynucleotide to enter a plant cell. In certain embodiments, an effective organosilicone compound can enable a polynucleotide to enter a plant cell in a manner permitting a polynucleotide mediated suppression of a target gene expression in the plant cell. In general, effective organosilicone compounds include, but are not limited to, compounds that can comprise: i) a trisiloxane head group that is covalently linked to, ii) an alkyl linker including, but not limited to, an n-propyl linker, that is covalently linked to, iii) a poly glycol chain, that is covalently linked to, iv) a terminal group. Trisiloxane head groups of such effective organosilicone compounds include, but are not limited to, heptamethyltrisiloxane. Alkyl linkers can include, but are not limited to, an n-propyl linker. Poly glycol chains include, but are not limited to, polyethylene glycol or polypropylene glycol. Poly glycol chains can comprise a mixture that provides an average chain length “n” of about “7.5”. In certain embodiments, the average chain length “n” can vary from about 5 to about 14. Terminal groups can include, but are not limited to, alkyl groups such as a methyl group. Effective organosilicone compounds are believed to include, but are not limited to, trisiloxane ethoxylate surfactants or polyalkylene oxide modified heptamethyl trisiloxane.
In certain embodiments, an organosilicone preparation that comprises an organosilicone compound comprising a trisiloxane head group is used in the methods and compositions provided herein. In certain embodiments, an organosilicone preparation that comprises an organosilicone compound comprising a heptamethyltrisiloxane head group is used in the methods and compositions provided herein. In certain embodiments, an organosilicone composition that comprises Compound I is used in the methods and compositions provided herein. In certain embodiments, an organosilicone composition that comprises Compound I is used in the methods and compositions provided herein. In certain embodiments of the methods and compositions provided herein, a composition that comprises a polynucleotide molecule and one or more effective organosilicone compound in the range of about 0.015 to about 2 percent by weight (wt percent) (e.g., about 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.5 wt percent) is used or provided.
Compositions of the present invention include but are not limited components that are one or more polynucleotides essentially identical to, or essentially complementary to a GS gene sequence (promoter, intron, exon, 5′ untranslated region, 3′ untranslated region), a transfer agent that provides for the polynucleotide to enter a plant cell, a herbicide that complements the action of the polynucleotide, one or more additional herbicides that further enhance the herbicide activity of the composition or provide an additional mode of action different from the complementing herbicide, various salts and stabilizing agents that enhance the utility of the composition as an admixture of the components of the composition.
Methods include one or more applications of a polynucleotide composition and one or more applications of a permeability-enhancing agent for conditioning of a plant to permeation by polynucleotides. When the agent for conditioning to permeation is an organosilicone composition or compound contained therein, embodiments of the polynucleotide molecules are double-stranded RNA oligonucleotides, single-stranded RNA oligonucleotides, double-stranded RNA polynucleotides, single-stranded RNA polynucleotides, double-stranded DNA oligonucleotides, single-stranded DNA oligonucleotides, double-stranded DNA polynucleotides, single-stranded DNA polynucleotides, chemically modified RNA or DNA oligonucleotides or polynucleotides or mixtures thereof.
Compositions and methods are useful for modulating the expression of an endogenous GS gene (for example, Pest Manag. Sci. 2009; 65: 216-222, GS249 mutants) or transgenic GS gene (for example, U.S. Pat. Nos. 7,910,805; 5,969,213; 5,489,520; 5,550,318; 5,874,265; 5,919,675; 5,561,236; 5,648,477; 5,646,024; 6,177,616; and 5,879,903) in a plant cell. In various embodiments, a GS gene includes coding (protein-coding or translatable) sequence, non-coding (non-translatable) sequence, or both coding and non-coding sequence. Compositions of the invention can include polynucleotides and oligonucleotides designed to target multiple genes, or multiple segments of one or more genes. The target gene can include multiple consecutive segments of a target gene, multiple non-consecutive segments of a target gene, multiple alleles of a target gene, or multiple target genes from one or more species.
One aspect is a method for modulating expression of a GS gene in a plant including (a) conditioning of a plant to permeation by polynucleotides and (b) treatment of the plant with the polynucleotide molecules, wherein the polynucleotide molecules include at least one segment of 18 or more contiguous nucleotides cloned from or otherwise identified from the target GS gene in either anti-sense or sense orientation, whereby the polynucleotide molecules permeate the interior of the plant and induce modulation of the target gene. The conditioning and polynucleotide application can be performed separately or in a single step. When the conditioning and polynucleotide application are performed in separate steps, the conditioning can precede or can follow the polynucleotide application within minutes, hours, or days. In some embodiments more than one conditioning step or more than one polynucleotide molecule application can be performed on the same plant. In embodiments of the method, the segment can be cloned or identified from (a) coding (protein-encoding), (b) non-coding (promoter and other gene related molecules), or (c) both coding and non-coding parts of the target gene. Non-coding parts include DNA, such as promoter regions or the RNA transcribed by the DNA that provide RNA regulatory molecules, including but not limited to: introns, 5′ or 3′ untranslated regions, and microRNAs (miRNA), trans-acting siRNAs, natural anti-sense siRNAs, and other small RNAs with regulatory function or RNAs having structural or enzymatic function including but not limited to: ribozymes, ribosomal RNAs, t-RNAs, aptamers, and riboswitches.
All publications, patents and patent applications are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.
The following examples are included to demonstrate examples of certain preferred embodiments. It should be appreciated by those of skill in the art that the techniques disclosed in the examples that follow represent approaches the inventors have found function well in the practice of the invention, and thus can be considered to constitute examples of preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments that are disclosed and still obtain a like or similar result without departing from the spirit and scope.
The target GS polynucleotide molecule naturally occurs in the genome of Abutilon theophrasti, Amaranthus albus, Amaranthus chlorostachys, Amaranthus graecizans, Amaranthus hybridus, Amaranthus lividus, Amaranthus palmeri, Amaranthus rudis, Amaranthus spinosus, Amaranthus thunbergii, Ambrosia trifida, Ambrosia artemisiifolia, Chenopodium album, Commelina diffusa, Convolvulus arvensis, Conyza canadensis, Lolium multiflorum, Euphorbia heterophylla, Kochia scoparia, Sorghum halepense, and Digitaria sanguinalis and include molecules related to the expression of a polypeptide identified as a GS, that include regulatory molecules, cDNAs comprising coding and noncoding regions of a GS gene and fragments thereof as shown in Table 1.
Polynucleotide molecules were extracted from these plant species by methods standard in the field, for example, total RNA is extracted using TRIZOL® reagent (Invitrogen Corp, Carlsbad, Calif., Cat. No. 15596-018; a monophasic solution of phenol and guanidine isothiocyanate), following the manufacturer's protocol or modifications thereof by those skilled in the art of polynucleotide extraction that may enhance recover or purity of the extracted RNA. Briefly, start with 1 gram of ground plant tissue for extraction. Prealiquot 10 milliliters (mL) TRIZOL® reagent to 15 mL conical tubes. Add ground powder to tubes and shake to homogenize. Incubate the homogenized samples for 5 minutes (min) at room temperature (RT) and then add 3 mL of chloroform. Shakes tubes vigorously by hand for 15-30 seconds (sec) and incubate at RT for 3 min. Centrifuge the tubes at 7,000 revolutions per minute (rpm) for 10 min at 4 degrees C. Transfer the aqueous phase to a new 1.5 mL tube and add 1 volume of cold isopropanol. Incubate the samples for 20-30 min at RT and centrifuge at 10,000 rpm for 10 min at 4 degrees C. Wash pellet with Sigma-grade 80 percent ethanol. Remove the supernatant and briefly air-dry the pellet. Dissolve the RNA pellet in approximately 200 microliters of DEPC treated water. Heat briefly at 65 degrees C. to dissolve pellet and vortex or pipet to resuspend RNA pellet. Adjust RNA concentration to 1-2 microgram/microliter.
DNA was extracted using EZNA SP Plant DNA Mini kit (Omega Biotek, Norcross Ga., Cat # D5511) and Lysing Matrix E tubes (Q-Biogen, Cat #6914), following the manufacturer's protocol or modifications thereof by those skilled in the art of polynucleotide extraction that may enhance recover or purity of the extracted DNA. Briefly, aliquot ground tissue to a Lysing Matrix E tube on dry ice, add 800 μl Buffer SP1 to each sample, homogenize in a bead beater for 35-45 sec, incubate on ice for 45-60 sec, centrifuge at >14000 rpm for 1 min at RT, add 10 microliter RNase A to the lysate, incubate at 65° C. for 10 min, centrifuge for 1 min at RT, add 280 μl Buffer SP2 and vortex to mix, incubate the samples on ice for 5 min, centrifuge at >10,000 g for 10 min at RT, transfer the supernatant to a homogenizer column in a 2 ml collection tube, centrifuge at 10,000 g for 2 min at RT, transfer the cleared lysate into a 1.5 ml microfuge tube, add 1.5 volumes Buffer SP3 to the cleared lysate, vortex immediately to obtain a homogeneous mixture, transfer up to 650 μl supernatant to the Hi-Bind column, centrifuge at 10,000 g for 1 min, repeat, apply 100 μl 65° C. Elution Buffer to the column, centrifuge at 10,000 g for 5 min at RT.
Next-generation DNA sequencers, such as the 454-FLX (Roche, Branford, Conn.), the SOLiD (Applied Biosystems,), and the Genome Analyzer (HiSeq2000, Illumina, San Diego, Calif.) were used to provide polynucleotide sequence from the DNA and RNA extracted from the plant tissues. Raw sequence data is assembled into contigs. The contig sequence is used to identify trigger molecules that can be applied to the plant to enable regulation of the gene expression.
The target DNA sequence isolated from genomic (gDNA) and coding DNA (cDNA) from the various weedy plant species for the GS gene and the assembled contigs were set forth in SEQ ID NOs: 1-59 and Table 1.
The gene sequences and fragments of Table 1 were divided into 200 polynucleotide (200-mer) lengths with 25 polynucleotide overlapping regions as in SEQ ID NOs: 37-1056. These polynucleotides are tested to select the most efficacious trigger regions across the length of any target sequence. The trigger polynucleotides are constructed as sense or anti-sense ssDNA or ssRNA, dsRNA, or dsDNA, or dsDNA/RNA hybrids and combined with an organosilicone based transfer agent to provide a polynucleotide preparation. The polynucleotides are combined into sets of two to three polynucleotides per set, using 4-8 nmol of each polynucleotide. Each polynucleotide set is prepared with the transfer agent and applied to a plant or a field of plants in combination with a GS inhibitor containing herbicide, or followed by a GS inhibitor treatment one to three days after the polynucleotide application, to determine the effect on the plant's susceptibility to a GS inhibitor. The effect is measured as stunting the growth and/or killing of the plant and is measured 8-14 days after treatment with the polynucleotide set and GS inhibitor. The most efficacious sets are identified and the individual polynucleotides are tested in the same methods as the sets are and the most efficacious single 200-mer identified. The 200-mer sequence is divided into smaller sequences of 50-70-mer regions with 10-15 polynucleotide overlapping regions and the polynucleotides tested individually. The most efficacious 50-70-mer is further divided into smaller sequences of 25-mer regions with a 12 to 13 polynucleotide overlapping region and tested for efficacy in combination with GS inhibitor treatment. By this method it is possible to identify an oligonucleotide or several oligonucleotides that are the most efficacious trigger molecule to effect plant sensitivity to a GS inhibitor or modulation of a GS gene expression. The modulation of GS gene expression is determined by the detection of GS siRNA molecules specific to a GS gene or by an observation of a reduction in the amount of GS RNA transcript produced relative to an untreated plant or by merely observing the anticipated phenotype of the application of the trigger with the GS inhibiting herbicide. Detection of siRNA can be accomplished, for example, using kits such as mirVana (Ambion, Austin Tex.) and mirPremier (Sigma-Aldrich, St Louis, Mo.).
The target DNA sequence isolated from genomic (gDNA) and coding DNA (cDNA) from the various weedy plant species for the GS gene and the assembled contigs w forth in SEQ ID NOs: 1-59 were divided into polynucleotide fragments as set forth in SEQ ID NOs: 60-1444.
The gene sequences and fragments of Table 1 were compared and 21-mers of contiguous polynucleotides were identified that had homology across the various GS gene sequences. The purpose is to identify trigger molecules that are useful as herbicidal molecules or in combination with a GS inhibitor herbicide across a broad range of weed species. The SEQ ID NOs: 1444-2045 sequences represent the 21-mers that are present in the GS gene of at least two of the weed species of Table 1. It is contemplated that additional 21-mers can be selected from the sequences of Table 1 that are specific for a single weed species or a few weeds species within a genus or trigger molecules that are at least 18 contiguous nucleotides, at least 19 contiguous nucleotides, at least 20 contiguous nucleotides or at least 21 contiguous nucleotides in length and at least 85 percent identical to a GS gene sequence selected from the group consisting of SEQ ID NOs: 1-59 or fragment thereof.
By this method it is possible to identify an oligonucleotide or several oligonucleotides that are the most efficacious trigger molecule to effect plant sensitivity to GS inhibitor or modulation of GS gene expression. The modulation of GS gene expression is determined by the detection of GS siRNA molecules specific to GS gene or by an observation of a reduction in the amount of GS RNA transcript produced relative to an untreated plant or by merely observing the anticipated phenotype of the application of the trigger with the GS inhibitor containing herbicide. Detection of siRNA can be accomplished, for example, using kits such as mirVana (Ambion, Austin Tex.) and mirPremier (Sigma-Aldrich, St Louis, Mo.).
The target DNA sequence isolated from genomic (gDNA) and coding DNA (cDNA) from the various weedy plant species for the GS gene and the assembled contigs as set forth in SEQ ID NOs: 1-59 were divided into fragments as set forth in SEQ ID NOs: 1444-2045.
Glyphosate-sensitive Palmer amaranth (A. palmeri R-22) plants were grown in the greenhouse (30/20 C day/night T; 14 hour photoperiod) in 4 inch square pots containing Sun Gro® Redi-Earth and 3.5 kg/cubic meter Osmocote® 14-14-14 fertilizer. Palmer amaranth plants at 5 to 10 cm in height were treated with a mixture of 5 short (40-mer) single-strand antisense single strand DNA polynucleotides (ssDNA) targeting GS coding sequence at 4 nm (nanomole) each, formulated in 20 millimolar sodium phosphate buffer (pH 6.8) containing 2 percent ammonium sulfate and 1 percent Silwet L-77. Three pools of ssDNA polynucleotides were tested. One pool named the GS CpGS1 contained 5 polynucleotides (SEQ ID NOs: 2046-2050) that were selected to target the gene encoding the chloroplastic targeted glutamine synthetase1 gene. The second pool named GS CytGS1 contained 5 polynucleotides (SEQ ID NOs: 2051-2055) that were selected to target the cytosolic glutamine synthetase1 gene. The third pool (GS Mix) was a combination of ssDNA trigger molecules, two each from the GS CpGS1 and GS CytGS1 pool plus one ssDNA (SEQ ID NOs: 2056) targeting a cytosolic glutamine synthetase2 gene. Plants are treated manually by pipetting 10 μL of polynucleotide solution on four fully expanded mature leaves, for a total of 40 microliters of solution per plant. Three days after the trigger polynucleotide treatment, the plants were treated with Ignite® rate (Bayer Cropscience) at 1/32× (23 grams/hectare) field rate. There were four replications of each treatment. Plant growth and development was visually rated sixteen days after herbicide treatment to determine the efficacy of the polynucleotide pool and herbicide treatments. The result shown in Table 2 as the average percent efficacy observed of the four replications relative to the untreated control.
A method to control weeds in a field comprises the use of trigger polynucleotides that can modulate the expression of a GS gene in one or more target weed plant species. In SEQ ID NOs: 1444-2045, an analysis of GS gene sequences from twenty-two plant species provided a collection of 21-mer polynucleotides that can be used in compositions to affect the growth or develop or sensitivity to GS inhibitor herbicide to control multiple weed species in a field. A composition containing 1 or 2 or 3 or 4 or more of the polynucleotides of SEQ ID NOs: 1444-2045 would enable broad activity of the composition against the multiple weed species that occur in a field environment.
The method includes creating a composition that comprises components that include at least one polynucleotide of SEQ ID NOs: 1444-2045 or any other effective gene expression modulating polynucleotide essentially identical or essentially complementary to SEQ ID NOs:1-59 or fragment thereof, a transfer agent that mobilizes the polynucleotide into a plant cell and a GS inhibiting herbicide and optionally a polynucleotide that modulates the expression of an essential gene and optionally a herbicide that has a different mode of action relative to a GS inhibitor. The polynucleotide of the composition includes a dsRNA, ssDNA or dsDNA or a combination thereof. A composition containing a polynucleotide can have a use rate of about 1 to 30 grams or more per acre depending on the size of the polynucleotide and the number of polynucleotides in the composition. The composition may include one or more additional herbicides as needed to provide effective multi-species weed control. A field of crop plants in need of weed plant control is treated by spray application of the composition. The composition can be provided as a tank mix, a sequential treatment of components (generally the polynucleotide followed by the herbicide), a simultaneous treatment or mixing of one or more of the components of the composition from separate containers. Treatment of the field can occur as often as needed to provide weed control and the components of the composition can be adjusted to target specific weed species or weed families.
Amaranthus palmeri
Amaranthus palmeri
Amaranthus palmeri
Amaranthus rudis
Amaranthus rudis
Amaranthus rudis
Amaranthus rudis
Ambrosia trifida
Ambrosia trifida
Ambrosia trifida
Conyza canadensis
Conyza canadensis
Conyza canadensis
Euphorbia heterophylla
Euphorbia heterophylla
Euphorbia heterophylla
Commelina diffusa
Commelina diffusa
Digitaria sanguinalis
Digitaria sanguinalis
Kochia scoparia
Kochia scoparia
Kochia scoparia
Kochia scoparia
Kochia scoparia
Kochia scoparia
Kochia scoparia
Lolium multiflorum
Lolium multiflorum
Lolium multiflorum
Lolium multiflorum
Lolium multiflorum
Lolium multiflorum
Lolium multiflorum
Lolium multiflorum
Lolium multiflorum
Abutilon theophrasti
Abutilon theophrasti
Abutilon theophrasti
Abutilon theophrasti
Amaranthus albus
Amaranthus chlorostachys
Amaranthus chlorostachys
Amaranthus graecizans
Amaranthus hybridus
Amaranthus lividus
Amaranthus spinosus
Amaranthus thunbergii
Amaranthus viridis
Euphorbia heterophylla
Sorghum halepense
Convolvulus arvensis
Chenopodium album
Ambrosia artemisiifolia
Euphorbia heterophylla
Euphorbia heterophylla
Euphorbia heterophylla
Euphorbia heterophylla
Euphorbia heterophylla
This application is a continuation-in-part application of U.S. application Ser. No. 13/612,948, filed on Sep. 13, 2012, which claims the benefit of priority of U.S. Provisional Application No. 61/534,076, filed on Sep. 13, 2011, both of which are incorporated in their entirety herein by reference.
| Number | Name | Date | Kind |
|---|---|---|---|
| 3687808 | Merigan et al. | Aug 1972 | A |
| 3791932 | Schuurs et al. | Feb 1974 | A |
| 3839153 | Schuurs et al. | Oct 1974 | A |
| 3850578 | McConnell | Nov 1974 | A |
| 3850752 | Schuurs et al. | Nov 1974 | A |
| 3853987 | Drcycr | Dec 1974 | A |
| 3867517 | Ling | Feb 1975 | A |
| 3879262 | Schuurs et al. | Apr 1975 | A |
| 3901654 | Gross | Aug 1975 | A |
| 3935074 | Rubenstein et al. | Jan 1976 | A |
| 3984533 | Uzgiris | Oct 1976 | A |
| 3996345 | Ullman et al. | Dec 1976 | A |
| 4034074 | Miles | Jul 1977 | A |
| 4098876 | Piasio et al. | Jul 1978 | A |
| 4469863 | Ts'o et al. | Sep 1984 | A |
| 4476301 | Imbach et al. | Oct 1984 | A |
| 4535060 | Comai | Aug 1985 | A |
| 4581847 | Hibberd et al. | Apr 1986 | A |
| 4666828 | Gusella | May 1987 | A |
| 4683202 | Mullis | Jul 1987 | A |
| 4761373 | Anderson et al. | Aug 1988 | A |
| 4769061 | Comai | Sep 1988 | A |
| 4801531 | Frossard | Jan 1989 | A |
| 4810648 | Stalker | Mar 1989 | A |
| 4879219 | Wands et al. | Nov 1989 | A |
| 4940835 | Shah et al. | Jul 1990 | A |
| 4971908 | Kishore et al. | Nov 1990 | A |
| 5004863 | Umbeck | Apr 1991 | A |
| 5011771 | Bellet et al. | Apr 1991 | A |
| 5013659 | Bedbrook et al. | May 1991 | A |
| 5015580 | Christou et al. | May 1991 | A |
| 5023243 | Tullis | Jun 1991 | A |
| 5034506 | Summcrton et al. | Jul 1991 | A |
| 5094945 | Comai | Mar 1992 | A |
| 5141870 | Bedbrook et al. | Aug 1992 | A |
| 5145783 | Kishorc et al. | Sep 1992 | A |
| 5159135 | Umbeck | Oct 1992 | A |
| 5166315 | Summerton et al. | Nov 1992 | A |
| 5177196 | Meyer, Jr. et al. | Jan 1993 | A |
| 5185444 | Summerlon et al. | Feb 1993 | A |
| 5188642 | Shah et al. | Feb 1993 | A |
| 5188897 | Suhadolnik et al. | Feb 1993 | A |
| 5192659 | Simons | Mar 1993 | A |
| 5214134 | Weis et al. | May 1993 | A |
| 5216141 | Benner | Jun 1993 | A |
| 5235033 | Summerlon et al. | Aug 1993 | A |
| 5264423 | Cohen et al. | Nov 1993 | A |
| 5264562 | Matteucci | Nov 1993 | A |
| 5264564 | Malleucci | Nov 1993 | A |
| 5272057 | Smulson et al. | Dec 1993 | A |
| 5276019 | Cohen et al. | Jan 1994 | A |
| 5281521 | Trojanowski et al. | Jan 1994 | A |
| 5286634 | Stadler et al. | Feb 1994 | A |
| 5286717 | Cohen et al. | Feb 1994 | A |
| 5304732 | Anderson et al. | Apr 1994 | A |
| 5310667 | Eichholtz et al. | May 1994 | A |
| 5312910 | Kishore et al. | May 1994 | A |
| 5321131 | Agrawal et al. | Jun 1994 | A |
| 5331107 | Anderson et al. | Jul 1994 | A |
| 5339107 | Henry et al. | Aug 1994 | A |
| 5346107 | Bouix et al. | Sep 1994 | A |
| 5378824 | Bedbrook et al. | Jan 1995 | A |
| 5384253 | Krzyzek et al. | Jan 1995 | A |
| 5390667 | Kumakura et al. | Feb 1995 | A |
| 5392910 | Bell et al. | Feb 1995 | A |
| 5393175 | Courville | Feb 1995 | A |
| 5399676 | Frochler | Mar 1995 | A |
| 5405938 | Summerlon et al. | Apr 1995 | A |
| 5405939 | Suhadolnik et al. | Apr 1995 | A |
| 5416011 | Hinchcc et al. | May 1995 | A |
| 5453496 | Caruthers et al. | Sep 1995 | A |
| 5455233 | Spielvogel et al. | Oct 1995 | A |
| 5459127 | Feigner et al. | Oct 1995 | A |
| 5460667 | Moriyuki et al. | Oct 1995 | A |
| 5462910 | Ito et al. | Oct 1995 | A |
| 5463174 | Moloney et al. | Oct 1995 | A |
| 5463175 | Barry et al. | Oct 1995 | A |
| 5466677 | Baxter et al. | Nov 1995 | A |
| 5470967 | Huie et al. | Nov 1995 | A |
| 5476925 | Letsinger et al. | Dec 1995 | A |
| 5489520 | Adams et al. | Feb 1996 | A |
| 5489677 | Sanghvi et al. | Feb 1996 | A |
| 5491288 | Chaubet et al. | Feb 1996 | A |
| 5510471 | Lebrun et al. | Apr 1996 | A |
| 5518908 | Corbin et al. | May 1996 | A |
| 5519126 | Hecht | May 1996 | A |
| 5536821 | Agrawal et al. | Jul 1996 | A |
| 5538880 | Lundquist et al. | Jul 1996 | A |
| 5541306 | Agrawal et al. | Jul 1996 | A |
| 5541307 | Cook et al. | Jul 1996 | A |
| 5550111 | Suhadolnik et al. | Aug 1996 | A |
| 5550318 | Adams et al. | Aug 1996 | A |
| 5550398 | Kocian et al. | Aug 1996 | A |
| 5550468 | Haberlein et al. | Aug 1996 | A |
| 5558071 | Ward et al. | Sep 1996 | A |
| 5561225 | Maddry et al. | Oct 1996 | A |
| 5561236 | Leemans et al. | Oct 1996 | A |
| 5563253 | Agrawal et al. | Oct 1996 | A |
| 5569834 | Hinchee et al. | Oct 1996 | A |
| 5571799 | Tkachuk et al. | Nov 1996 | A |
| 5587361 | Cook et al. | Dec 1996 | A |
| 5591616 | Hiei et al. | Jan 1997 | A |
| 5593874 | Brown et al. | Jan 1997 | A |
| 5596086 | Matteucci et al. | Jan 1997 | A |
| 5597717 | Guerineau et al. | Jan 1997 | A |
| 5602240 | De Mesmaeker et al. | Feb 1997 | A |
| 5605011 | Bedbrook et al. | Feb 1997 | A |
| 5608046 | Cook et al. | Mar 1997 | A |
| 5610289 | Cook et al. | Mar 1997 | A |
| 5618704 | Sanghvi et al. | Apr 1997 | A |
| 5623070 | Cook et al. | Apr 1997 | A |
| 5625050 | Beaton et al. | Apr 1997 | A |
| 5627061 | Barry et al. | May 1997 | A |
| 5633360 | Bischofberger et al. | May 1997 | A |
| 5633435 | Barry et al. | May 1997 | A |
| 5633448 | Lebrun et al. | May 1997 | A |
| 5639024 | Mueller et al. | Jun 1997 | A |
| 5646024 | Leemans et al. | Jul 1997 | A |
| 5648477 | Leemans et al. | Jul 1997 | A |
| 5663312 | Chaturvedula | Sep 1997 | A |
| 5677437 | Teng et al. | Oct 1997 | A |
| 5677439 | Weis et al. | Oct 1997 | A |
| 5719046 | Guerineau et al. | Feb 1998 | A |
| 5721138 | Lawn | Feb 1998 | A |
| 5731180 | Dietrich | Mar 1998 | A |
| 5739180 | Taylor-Smith | Apr 1998 | A |
| 5746180 | Jefferson et al. | May 1998 | A |
| 5767361 | Dietrich | Jun 1998 | A |
| 5767373 | Ward et al. | Jun 1998 | A |
| 5780708 | Lundquist et al. | Jul 1998 | A |
| 5804425 | Barry et al. | Sep 1998 | A |
| 5824877 | Hinchee et al. | Oct 1998 | A |
| 5837848 | Ely et al. | Nov 1998 | A |
| 5859347 | Brown et al. | Jan 1999 | A |
| 5866775 | Eichholtz et al. | Feb 1999 | A |
| 5874265 | Adams et al. | Feb 1999 | A |
| 5879903 | Strauch et al. | Mar 1999 | A |
| 5914451 | Martinell et al. | Jun 1999 | A |
| 5919675 | Adams et al. | Jul 1999 | A |
| 5928937 | Kakefuda et al. | Jul 1999 | A |
| 5939602 | Volrath et al. | Aug 1999 | A |
| 5969213 | Adams et al. | Oct 1999 | A |
| 5981840 | Zhao et al. | Nov 1999 | A |
| 5985793 | Sandbrink | Nov 1999 | A |
| RE36449 | Lebrun et al. | Dec 1999 | E |
| 6040497 | Spencer et al. | Mar 2000 | A |
| 6056938 | Unger et al. | May 2000 | A |
| 6069115 | Pallett et al. | May 2000 | A |
| 6084089 | Mine et al. | Jul 2000 | A |
| 6084155 | Volrath et al. | Jul 2000 | A |
| 6118047 | Anderson et al. | Sep 2000 | A |
| 6121513 | Zhang et al. | Sep 2000 | A |
| 6130366 | Herrera-Estrella et al. | Oct 2000 | A |
| 6140078 | Sanders et al. | Oct 2000 | A |
| 6153812 | Fry et al. | Nov 2000 | A |
| 6160208 | Lundquist et al. | Dec 2000 | A |
| 6177616 | Bartsch et al. | Jan 2001 | B1 |
| 6194636 | McElroy et al. | Feb 2001 | B1 |
| 6225105 | Sathasivan et al. | May 2001 | B1 |
| 6225114 | Eichholtz et al. | May 2001 | B1 |
| 6232526 | McElroy et al. | May 2001 | B1 |
| 6245968 | Boudec et al. | Jun 2001 | B1 |
| 6248876 | Barry et al. | Jun 2001 | B1 |
| 6252138 | Karimi et al. | Jun 2001 | B1 |
| RE37287 | Lebrun et al. | Jul 2001 | E |
| 6268549 | Sailland et al. | Jul 2001 | B1 |
| 6271359 | Norris et al. | Aug 2001 | B1 |
| 6282837 | Ward et al. | Sep 2001 | B1 |
| 6288306 | Ward et al. | Sep 2001 | B1 |
| 6288312 | Christou et al. | Sep 2001 | B1 |
| 6294714 | Matsunaga et al. | Sep 2001 | B1 |
| 6326193 | Liu et al. | Dec 2001 | B1 |
| 6329571 | Hiei | Dec 2001 | B1 |
| 6348185 | Piwnica-Worms | Feb 2002 | B1 |
| 6365807 | Christou et al. | Apr 2002 | B1 |
| 6384301 | Martinell et al. | May 2002 | B1 |
| 6385902 | Schipper et al. | May 2002 | B1 |
| 6399861 | Anderson et al. | Jun 2002 | B1 |
| 6403865 | Koziel et al. | Jun 2002 | B1 |
| 6414222 | Gengenbach et al. | Jul 2002 | B1 |
| 6421956 | Boukens et al. | Jul 2002 | B1 |
| 6426446 | McElroy et al. | Jul 2002 | B1 |
| 6433252 | Kriz et al. | Aug 2002 | B1 |
| 6437217 | McElroy et al. | Aug 2002 | B1 |
| 6453609 | Soll et al. | Sep 2002 | B1 |
| 6479291 | Kumagai et al. | Nov 2002 | B2 |
| 6506559 | Fire et al. | Jan 2003 | B1 |
| 6506599 | Yoon | Jan 2003 | B1 |
| 6642435 | Rafalski et al. | Nov 2003 | B1 |
| 6644341 | Chemo et al. | Nov 2003 | B1 |
| 6645914 | Woznica et al. | Nov 2003 | B1 |
| 6768044 | Boudcc et al. | Jul 2004 | B1 |
| 6992237 | Habben et al. | Jan 2006 | B1 |
| 7022896 | Weeks et al. | Apr 2006 | B1 |
| 7026528 | Cheng et al. | Apr 2006 | B2 |
| RE39247 | Barry et al. | Aug 2006 | E |
| 7105724 | Weeks et al. | Sep 2006 | B2 |
| 7119256 | Shimizu et al. | Oct 2006 | B2 |
| 7138564 | Tian et al. | Nov 2006 | B2 |
| 7297541 | Moshiri et al. | Nov 2007 | B2 |
| 7304209 | Zink et al. | Dec 2007 | B2 |
| 7312379 | Andrews et al. | Dec 2007 | B2 |
| 7323310 | Peters et al. | Jan 2008 | B2 |
| 7371927 | Yao et al. | May 2008 | B2 |
| 7392379 | Le Pennec et al. | Jun 2008 | B2 |
| 7405347 | Hammer et al. | Jul 2008 | B2 |
| 7406981 | Hemo et al. | Aug 2008 | B2 |
| 7462379 | Fukuda et al. | Dec 2008 | B2 |
| 7485777 | Nakajima et al. | Feb 2009 | B2 |
| 7525013 | Hildebrand | Apr 2009 | B2 |
| 7550578 | Budworth et al. | Jun 2009 | B2 |
| 7622301 | Ren et al. | Nov 2009 | B2 |
| 7657299 | Huizenga et al. | Feb 2010 | B2 |
| 7671254 | Trancl et al. | Mar 2010 | B2 |
| 7714188 | Castle et al. | May 2010 | B2 |
| 7738626 | Weese et al. | Jun 2010 | B2 |
| 7807791 | Sekar et al. | Oct 2010 | B2 |
| 7838263 | Dam et al. | Nov 2010 | B2 |
| 7838733 | Wright et al. | Nov 2010 | B2 |
| 7842856 | Tranel et al. | Nov 2010 | B2 |
| 7884262 | Clemente et al. | Feb 2011 | B2 |
| 7910805 | Duck et al. | Mar 2011 | B2 |
| 7935869 | Pallett et al. | May 2011 | B2 |
| 7943819 | Baum et al. | May 2011 | B2 |
| 7973218 | McCutchen et al. | Jul 2011 | B2 |
| 8090164 | Bullitt et al. | Jan 2012 | B2 |
| 8143480 | Axtell et al. | Mar 2012 | B2 |
| 8226938 | Meikle et al. | Jul 2012 | B1 |
| 8548778 | Hart et al. | Oct 2013 | B1 |
| 8554490 | Tang et al. | Oct 2013 | B2 |
| 9121022 | Sammons et al. | Sep 2015 | B2 |
| 9422557 | Ader | Aug 2016 | B2 |
| 9445603 | Baum et al. | Sep 2016 | B2 |
| 9777288 | Beattie et al. | Oct 2017 | B2 |
| 9850496 | Beattie et al. | Dec 2017 | B2 |
| 9856495 | Beattie et al. | Oct 2018 | B2 |
| 20010006797 | Kumagai et al. | Jul 2001 | A1 |
| 20010042257 | Connor-Ward et al. | Nov 2001 | A1 |
| 20020069430 | Kiaska et al. | Jun 2002 | A1 |
| 20020106653 | Kurane et al. | Aug 2002 | A1 |
| 20020114784 | Li et al. | Aug 2002 | A1 |
| 20030150017 | Mesa et al. | Aug 2003 | A1 |
| 20030154508 | Stevens et al. | Aug 2003 | A1 |
| 20030167537 | Jiang | Sep 2003 | A1 |
| 20030221211 | Rottmann et al. | Nov 2003 | A1 |
| 20040029275 | Brown et al. | Feb 2004 | A1 |
| 20040053289 | Christian et al. | Mar 2004 | A1 |
| 20040055041 | Labate et al. | Mar 2004 | A1 |
| 20040072692 | Hoffman et al. | Apr 2004 | A1 |
| 20040082475 | Hoffman et al. | Apr 2004 | A1 |
| 20040123347 | Hinchey et al. | Jun 2004 | A1 |
| 20040126845 | Eenennaam et al. | Jul 2004 | A1 |
| 20040133944 | Hake et al. | Jul 2004 | A1 |
| 20040147475 | Li et al. | Jul 2004 | A1 |
| 20040177399 | Hammer et al. | Sep 2004 | A1 |
| 20040216189 | Houmard et al. | Oct 2004 | A1 |
| 20040244075 | Cai et al. | Dec 2004 | A1 |
| 20040250310 | Shukla et al. | Dec 2004 | A1 |
| 20050005319 | della-Cioppa et al. | Jan 2005 | A1 |
| 20050044591 | Yao | Feb 2005 | A1 |
| 20050215435 | Menges et al. | Sep 2005 | A1 |
| 20050223425 | Clinton et al. | Oct 2005 | A1 |
| 20050246784 | Plesch et al. | Nov 2005 | A1 |
| 20050250647 | Hills et al. | Nov 2005 | A1 |
| 20050289664 | Moshiri et al. | Dec 2005 | A1 |
| 20060009358 | Kibler et al. | Jan 2006 | A1 |
| 20060021087 | Baum et al. | Jan 2006 | A1 |
| 20060040826 | Eaton et al. | Feb 2006 | A1 |
| 20060111241 | Gerwick, III et al. | May 2006 | A1 |
| 20060130172 | Whaley et al. | Jun 2006 | A1 |
| 20060135758 | Wu | Jun 2006 | A1 |
| 20060200878 | Lutfiyya et al. | Sep 2006 | A1 |
| 20060223708 | Hoffman et al. | Oct 2006 | A1 |
| 20060223709 | Helmke et al. | Oct 2006 | A1 |
| 20060247197 | Van De Craen et al. | Nov 2006 | A1 |
| 20060272049 | Waterhouse et al. | Nov 2006 | A1 |
| 20060276339 | Windsor et al. | Dec 2006 | A1 |
| 20070011775 | Allen et al. | Jan 2007 | A1 |
| 20070021360 | Nyce et al. | Jan 2007 | A1 |
| 20070050863 | Tranel et al. | Mar 2007 | A1 |
| 20070124836 | Baum et al. | May 2007 | A1 |
| 20070199095 | Allen et al. | Aug 2007 | A1 |
| 20070250947 | Boukharov et al. | Oct 2007 | A1 |
| 20070259785 | Heck et al. | Nov 2007 | A1 |
| 20070269815 | Rivory et al. | Nov 2007 | A1 |
| 20070281900 | Cui et al. | Dec 2007 | A1 |
| 20070300329 | Allen et al. | Dec 2007 | A1 |
| 20080022423 | Roberts et al. | Jan 2008 | A1 |
| 20080050342 | Fire et al. | Feb 2008 | A1 |
| 20080092256 | Kohn | Apr 2008 | A1 |
| 20080113351 | Naito et al. | May 2008 | A1 |
| 20080155716 | Sonnewald et al. | Jun 2008 | A1 |
| 20080214443 | Baum et al. | Sep 2008 | A1 |
| 20090011934 | Zawicrucha et al. | Jan 2009 | A1 |
| 20090018016 | Duck et al. | Jan 2009 | A1 |
| 20090036311 | Witschel et al. | Feb 2009 | A1 |
| 20090054240 | Witschel et al. | Feb 2009 | A1 |
| 20090075921 | Ikegawa et al. | Mar 2009 | A1 |
| 20090094717 | Troukhan et al. | Apr 2009 | A1 |
| 20090098614 | Zamorc et al. | Apr 2009 | A1 |
| 20090118214 | Paldi et al. | May 2009 | A1 |
| 20090137395 | Chicoine et al. | May 2009 | A1 |
| 20090144848 | Kovalic et al. | Jun 2009 | A1 |
| 20090165153 | Wang et al. | Jun 2009 | A1 |
| 20090165166 | Feng et al. | Jun 2009 | A1 |
| 20090205079 | Kumar et al. | Aug 2009 | A1 |
| 20090215628 | Witschel et al. | Aug 2009 | A1 |
| 20090285784 | Raemaekers et al. | Nov 2009 | A1 |
| 20090293148 | Ren et al. | Nov 2009 | A1 |
| 20090298787 | Raemaekers et al. | Dec 2009 | A1 |
| 20090306189 | Raemaekers et al. | Dec 2009 | A1 |
| 20090307803 | Baum et al. | Dec 2009 | A1 |
| 20100005551 | Roberts et al. | Jan 2010 | A1 |
| 20100048670 | Biard et al. | Feb 2010 | A1 |
| 20100068172 | Van De Craen | Mar 2010 | A1 |
| 20100071088 | Sela et al. | Mar 2010 | A1 |
| 20100099561 | Selby et al. | Apr 2010 | A1 |
| 20100100988 | Tranel et al. | Apr 2010 | A1 |
| 20100152443 | Hirai et al. | Jun 2010 | A1 |
| 20100154083 | Ross et al. | Jun 2010 | A1 |
| 20100192237 | Ren et al. | Jul 2010 | A1 |
| 20100247578 | Salama | Sep 2010 | A1 |
| 20100248373 | Baba et al. | Sep 2010 | A1 |
| 20110015084 | Christian et al. | Jan 2011 | A1 |
| 20110015284 | Dees et al. | Jan 2011 | A1 |
| 20110028412 | Cappello et al. | Feb 2011 | A1 |
| 20110035836 | Endes et al. | Feb 2011 | A1 |
| 20110041400 | Trias Vila et al. | Feb 2011 | A1 |
| 20110053226 | Rohayem | Mar 2011 | A1 |
| 20110098180 | Michel et al. | Apr 2011 | A1 |
| 20110105327 | Nelson | May 2011 | A1 |
| 20110105329 | Song et al. | May 2011 | A1 |
| 20110112570 | Mannava et al. | May 2011 | A1 |
| 20110126310 | Feng et al. | May 2011 | A1 |
| 20110126311 | Velcheva et al. | May 2011 | A1 |
| 20110152339 | Brown et al. | Jun 2011 | A1 |
| 20110152346 | Karleson et al. | Jun 2011 | A1 |
| 20110152353 | Koizumi et al. | Jun 2011 | A1 |
| 20110160082 | Woo et al. | Jun 2011 | A1 |
| 20110166022 | Isracls et al. | Jul 2011 | A1 |
| 20110166023 | Nettleton-Hammond et al. | Jul 2011 | A1 |
| 20110171176 | Baas et al. | Jul 2011 | A1 |
| 20110171287 | Saarma et al. | Jul 2011 | A1 |
| 20110177949 | Krapp et al. | Jul 2011 | A1 |
| 20110185444 | Li et al. | Jul 2011 | A1 |
| 20110185445 | Bogner et al. | Jul 2011 | A1 |
| 20110191897 | Poree et al. | Aug 2011 | A1 |
| 20110201501 | Song et al. | Aug 2011 | A1 |
| 20110203013 | Peterson et al. | Aug 2011 | A1 |
| 20110296555 | Ivashuta et al. | Dec 2011 | A1 |
| 20110296556 | Sammons et al. | Dec 2011 | A1 |
| 20120036594 | Cardoza et al. | Feb 2012 | A1 |
| 20120107355 | Harris et al. | May 2012 | A1 |
| 20120108497 | Paldi et al. | May 2012 | A1 |
| 20120137387 | Baum et al. | May 2012 | A1 |
| 20120150048 | Kang et al. | Jun 2012 | A1 |
| 20120156784 | Adams, Jr. et al. | Jun 2012 | A1 |
| 20120157512 | Ben-Chanoch et al. | Jun 2012 | A1 |
| 20120164205 | Baum et al. | Jun 2012 | A1 |
| 20120174262 | Azhakanandam et al. | Jul 2012 | A1 |
| 20120185967 | Sela et al. | Jul 2012 | A1 |
| 20120198586 | Narva et al. | Aug 2012 | A1 |
| 20120230565 | Steinberg et al. | Sep 2012 | A1 |
| 20120258646 | Sela et al. | Oct 2012 | A1 |
| 20130003213 | Kabelac et al. | Jan 2013 | A1 |
| 20130041004 | Drager et al. | Feb 2013 | A1 |
| 20130047297 | Sammons et al. | Feb 2013 | A1 |
| 20130047298 | Tang | Feb 2013 | A1 |
| 20130060133 | Kassab et al. | Mar 2013 | A1 |
| 20130067618 | Ader et al. | Mar 2013 | A1 |
| 20130084243 | Goetsch et al. | Apr 2013 | A1 |
| 20130096073 | Sidelman | Apr 2013 | A1 |
| 20130097726 | Ader et al. | Apr 2013 | A1 |
| 20130212739 | Giritch et al. | Aug 2013 | A1 |
| 20130226003 | Edic et al. | Aug 2013 | A1 |
| 20130254940 | Ader et al. | Sep 2013 | A1 |
| 20130254941 | Ader et al. | Sep 2013 | A1 |
| 20130288895 | Ader et al. | Oct 2013 | A1 |
| 20130318657 | Avniel et al. | Nov 2013 | A1 |
| 20130318658 | Ader et al. | Nov 2013 | A1 |
| 20130324842 | Mittal et al. | Dec 2013 | A1 |
| 20130326731 | Ader et al. | Dec 2013 | A1 |
| 20140018241 | Sammons et al. | Jan 2014 | A1 |
| 20140057789 | Sammons et al. | Feb 2014 | A1 |
| 20140109258 | Van De Craen et al. | Apr 2014 | A1 |
| 20140230090 | Avniel et al. | Aug 2014 | A1 |
| 20140274712 | Finnessy et al. | Sep 2014 | A1 |
| 20140275208 | Hu et al. | Sep 2014 | A1 |
| 20140296503 | Avniel et al. | Oct 2014 | A1 |
| 20150096079 | Avniel et al. | Apr 2015 | A1 |
| 20150143580 | Beattie et al. | May 2015 | A1 |
| 20150159156 | Tnberg et al. | Jun 2015 | A1 |
| 20150203867 | Beattie et al. | Jul 2015 | A1 |
| 20150240258 | Beattie et al. | Aug 2015 | A1 |
| 20160015035 | Tao | Jan 2016 | A1 |
| 20160029644 | Tao | Feb 2016 | A1 |
| Number | Date | Country |
|---|---|---|
| 2008258254 | Jul 2014 | AU |
| 20 14262189 | Nov 2014 | AU |
| 101279950 | Oct 2008 | CN |
| 101279951 | Oct 2008 | CN |
| 101892247 | Nov 2010 | CN |
| 101892247 | Nov 2010 | CN |
| 101914540 | Dec 2010 | CN |
| 102154364 | Aug 2011 | CN |
| 102481311 | May 2012 | CN |
| 102822350 | Dec 2012 | CN |
| 102906263 | Jan 2013 | CN |
| 288618 | Apr 1991 | DE |
| 10000600 | Jul 2001 | DE |
| 10116399 | Oct 2002 | DE |
| 10256353 | Jun 2003 | DE |
| 10256354 | Jun 2003 | DE |
| 10256367 | Jun 2003 | DE |
| 10204951 | Aug 2003 | DE |
| 10234875 | Feb 2004 | DE |
| 10234876 | Feb 2004 | DE |
| 102004054666 | May 2006 | DE |
| 102005014638 | Oct 2006 | DE |
| 102005014906 | Oct 2006 | DE |
| 102007012168 | Sep 2008 | DE |
| 102010042866 | May 2011 | DE |
| 0 804 600 | Nov 1997 | EP |
| 1 155 615 | Nov 2001 | EP |
| 1 157 991 | Nov 2001 | EP |
| 1 238 586 | Sep 2002 | EP |
| 1 416 049 | May 2004 | EP |
| 1 496 123 | Jan 2005 | EP |
| 1 889 902 | Feb 2008 | EP |
| 1 964 919 | Sep 2008 | EP |
| 2 147 919 | Jan 2010 | EP |
| 2 160 098 | Nov 2010 | EP |
| 2 530 159 | Mar 2011 | EP |
| 2 305 813 | Apr 2011 | EP |
| 2 473 024 | Jul 2012 | EP |
| 2 545 182 | Jan 2013 | EP |
| 2001253874 | Sep 2001 | JP |
| 2002080454 | Mar 2002 | JP |
| 2002138075 | May 2002 | JP |
| 2002145707 | May 2002 | JP |
| 2002220389 | Aug 2002 | JP |
| 2003064059 | Mar 2003 | JP |
| 2003096059 | Apr 2003 | JP |
| 2004051628 | Feb 2004 | JP |
| 2004107228 | Apr 2004 | JP |
| 2005008583 | Jan 2005 | JP |
| 2005239675 | Sep 2005 | JP |
| 2005314407 | Nov 2005 | JP |
| 2006232824 | Sep 2006 | JP |
| 2006282552 | Oct 2006 | JP |
| 2007153847 | Jun 2007 | JP |
| 2007161701 | Jun 2007 | JP |
| 2007182404 | Jul 2007 | JP |
| 2008074840 | Apr 2008 | JP |
| 2008074841 | Apr 2008 | JP |
| 2008133207 | Jun 2008 | JP |
| 2008133218 | Jun 2008 | JP |
| 2008169121 | Jul 2008 | JP |
| 2009-508481 | Mar 2009 | JP |
| 2009067739 | Apr 2009 | JP |
| 2009114128 | May 2009 | JP |
| 2009126792 | Jun 2009 | JP |
| 2009137851 | Jun 2009 | JP |
| 2016-532440 | Oct 2015 | JP |
| 2 291 613 | Jan 2007 | RU |
| 2 337 529 | Nov 2008 | RU |
| WO 8911789 | Dec 1989 | WO |
| WO 9534659 | Dec 1995 | WO |
| WO 9534668 | Dec 1995 | WO |
| WO 96005721 | Feb 1996 | WO |
| WO 96033270 | Oct 1996 | WO |
| WO 96038567 | Dec 1996 | WO |
| WO 96040964 | Dec 1996 | WO |
| WO 9749816 | Dec 1997 | WO |
| WO 9914348 | Mar 1999 | WO |
| WO 99024585 | May 1999 | WO |
| WO 9926467 | Jun 1999 | WO |
| WO 9927116 | Jun 1999 | WO |
| WO 9932619 | Jul 1999 | WO |
| WO 9961631 | Dec 1999 | WO |
| WO 9967367 | Dec 1999 | WO |
| WO 0032757 | Jun 2000 | WO |
| WO 00044914 | Aug 2000 | WO |
| WO 200107601 | Feb 2001 | WO |
| WO 2001085970 | Nov 2001 | WO |
| WO 0214472 | Feb 2002 | WO |
| WO 02066660 | Aug 2002 | WO |
| WO 03000679 | Jan 2003 | WO |
| WO 03004649 | Jan 2003 | WO |
| WO 03006422 | Jan 2003 | WO |
| WO 2003004649 | Jan 2003 | WO |
| WO 03012052 | Feb 2003 | WO |
| WO 03013247 | Feb 2003 | WO |
| WO 03016308 | Feb 2003 | WO |
| WO 2003014357 | Feb 2003 | WO |
| WO 03020704 | Mar 2003 | WO |
| WO 03022051 | Mar 2003 | WO |
| WO 03022831 | Mar 2003 | WO |
| WO 03022843 | Mar 2003 | WO |
| WO 03029243 | Apr 2003 | WO |
| WO 03037085 | May 2003 | WO |
| WO 03037878 | May 2003 | WO |
| WO 03045878 | Jun 2003 | WO |
| WO 03050087 | Jun 2003 | WO |
| WO 03051823 | Jun 2003 | WO |
| WO 03051824 | Jun 2003 | WO |
| WO 03051846 | Jun 2003 | WO |
| WO 03064625 | Aug 2003 | WO |
| WO 03076409 | Sep 2003 | WO |
| WO 03077648 | Sep 2003 | WO |
| WO 03087067 | Oct 2003 | WO |
| WO 03090539 | Nov 2003 | WO |
| WO 03091217 | Nov 2003 | WO |
| WO 03093269 | Nov 2003 | WO |
| WO 03104206 | Dec 2003 | WO |
| WO 2004002947 | Jan 2004 | WO |
| WO 2004002981 | Jan 2004 | WO |
| WO 2004005485 | Jan 2004 | WO |
| WO 2004009761 | Jan 2004 | WO |
| WO 2004011429 | Feb 2004 | WO |
| WO 2004022771 | Mar 2004 | WO |
| WO 2004029060 | Apr 2004 | WO |
| WO 2004035545 | Apr 2004 | WO |
| WO 2004035563 | Apr 2004 | WO |
| WO 2004035564 | Apr 2004 | WO |
| WO 2004037787 | May 2004 | WO |
| WO 2004049806 | Jun 2004 | WO |
| WO 2004062351 | Jul 2004 | WO |
| WO 2004067518 | Aug 2004 | WO |
| WO 2004067527 | Aug 2004 | WO |
| WO 2004074443 | Sep 2004 | WO |
| WO 2004077950 | Sep 2004 | WO |
| WO 2005000824 | Jan 2005 | WO |
| WO 2005003362 | Jan 2005 | WO |
| WO 2005007627 | Jan 2005 | WO |
| WO 2005007860 | Jan 2005 | WO |
| WO 2005040152 | May 2005 | WO |
| WO 2005047233 | May 2005 | WO |
| WO 2005047281 | May 2005 | WO |
| WO 2005061443 | Jul 2005 | WO |
| WO 2005061464 | Jul 2005 | WO |
| WO 2005068434 | Jul 2005 | WO |
| WO 2005070889 | Aug 2005 | WO |
| WO 2005089551 | Sep 2005 | WO |
| WO 2005095335 | Oct 2005 | WO |
| WO 2005107437 | Nov 2005 | WO |
| WO 2005110068 | Nov 2005 | WO |
| WO 2006006569 | Jan 2006 | WO |
| WO 2006024820 | Mar 2006 | WO |
| WO 2006029828 | Mar 2006 | WO |
| WO 2006029829 | Mar 2006 | WO |
| WO 2006037945 | Apr 2006 | WO |
| WO 2006050803 | May 2006 | WO |
| WO 2006074400 | Jul 2006 | WO |
| WO 2006090792 | Aug 2006 | WO |
| WO 2006123088 | Nov 2006 | WO |
| WO 2006125687 | Nov 2006 | WO |
| WO 2006125688 | Nov 2006 | WO |
| WO 2006132270 | Dec 2006 | WO |
| WO 2006138638 | Dec 2006 | WO |
| WO 2007003294 | Jan 2007 | WO |
| WO 2007007316 | Jan 2007 | WO |
| WO 2007024783 | Mar 2007 | WO |
| WO 2007026834 | Mar 2007 | WO |
| WO 2007035650 | Mar 2007 | WO |
| WO 2007038788 | Apr 2007 | WO |
| WO 2007039454 | Apr 2007 | WO |
| WO 2007050715 | May 2007 | WO |
| WO 2007051462 | May 2007 | WO |
| WO 2007070389 | Jun 2007 | WO |
| WO 2007071900 | Jun 2007 | WO |
| WO 2007074405 | Jul 2007 | WO |
| WO 2007077201 | Jul 2007 | WO |
| WO 2007077247 | Jul 2007 | WO |
| WO 2007080126 | Jul 2007 | WO |
| WO 2007080127 | Jul 2007 | WO |
| WO 2007083193 | Jul 2007 | WO |
| WO 2007096576 | Aug 2007 | WO |
| WO 2007051462 | Oct 2007 | WO |
| WO 2007119434 | Oct 2007 | WO |
| WO 2007134984 | Nov 2007 | WO |
| WO 2008007100 | Jan 2008 | WO |
| WO 2008009908 | Jan 2008 | WO |
| WO 2008029084 | Mar 2008 | WO |
| WO 2008042231 | Apr 2008 | WO |
| WO 2008059948 | May 2008 | WO |
| WO 2008063203 | May 2008 | WO |
| WO 2008071918 | Jun 2008 | WO |
| WO 2008074991 | Jun 2008 | WO |
| WO 2008084073 | Jul 2008 | WO |
| WO 2008100426 | Aug 2008 | WO |
| WO 2008102908 | Aug 2008 | WO |
| WO 2008148223 | Dec 2008 | WO |
| WO 2008152072 | Dec 2008 | WO |
| WO 2008152073 | Dec 2008 | WO |
| WO 2009000757 | Dec 2008 | WO |
| WO 2009005297 | Jan 2009 | WO |
| WO 2009029690 | Mar 2009 | WO |
| WO 2009035150 | Mar 2009 | WO |
| WO 2009037329 | Mar 2009 | WO |
| WO 2009046384 | Apr 2009 | WO |
| WO 2009060429 | May 2009 | WO |
| WO 2009063180 | May 2009 | WO |
| WO 2009068170 | Jun 2009 | WO |
| WO 2009068171 | Jun 2009 | WO |
| WO 2009086041 | Jul 2009 | WO |
| WO 2009090401 | Jul 2009 | WO |
| WO 2009090402 | Jul 2009 | WO |
| WO 2009115788 | Sep 2009 | WO |
| WO 2009116558 | Sep 2009 | WO |
| WO 2009125401 | Oct 2009 | WO |
| WO 2009144079 | Dec 2009 | WO |
| WO 2009152995 | Dec 2009 | WO |
| WO 2009153607 | Dec 2009 | WO |
| WO 2009158258 | Dec 2009 | WO |
| WO 2010012649 | Feb 2010 | WO |
| WO 2010026989 | Mar 2010 | WO |
| WO 2010034153 | Apr 2010 | WO |
| WO 2010049270 | May 2010 | WO |
| WO 2010049369 | May 2010 | WO |
| WO 2010049405 | May 2010 | WO |
| WO 2010049414 | May 2010 | WO |
| WO 2010056519 | May 2010 | WO |
| WO 2010063422 | Jun 2010 | WO |
| WO 2010069802 | Jun 2010 | WO |
| WO 2010078906 | Jul 2010 | WO |
| WO 2010078912 | Jul 2010 | WO |
| WO 2010093788 | Aug 2010 | WO |
| WO 2010104217 | Sep 2010 | WO |
| WO 2010108611 | Sep 2010 | WO |
| WO 2010112826 | Oct 2010 | WO |
| WO 2010116122 | Oct 2010 | WO |
| WO 2010119906 | Oct 2010 | WO |
| WO 2010130970 | Nov 2010 | WO |
| WO 2011001434 | Jan 2011 | WO |
| WO 2011003776 | Jan 2011 | WO |
| WO 2011028836 | Mar 2011 | WO |
| WO 2011035874 | Mar 2011 | WO |
| WO 2011045796 | Apr 2011 | WO |
| WO 2011065451 | Jun 2011 | WO |
| WO 2011067745 | Jun 2011 | WO |
| WO 2011075188 | Jun 2011 | WO |
| WO 2011080674 | Jul 2011 | WO |
| WO 2011112570 | Sep 2011 | WO |
| WO 2011132127 | Oct 2011 | WO |
| WO 2012001626 | Jan 2012 | WO |
| WO 2012056401 | May 2012 | WO |
| WO 2012092580 | Jul 2012 | WO |
| WO 2012156342 | Nov 2012 | WO |
| WO 2012164100 | Dec 2012 | WO |
| WO 2013010691 | Jan 2013 | WO |
| WO 2013025670 | Feb 2013 | WO |
| WO 2013039990 | Mar 2013 | WO |
| WO 2013040005 | Mar 2013 | WO |
| WO 2013040021 | Mar 2013 | WO |
| WO 2013040033 | Mar 2013 | WO |
| WO 2013040049 | Mar 2013 | WO |
| WO 2013040116 | Mar 2013 | WO |
| WO 2013040117 | Mar 2013 | WO |
| WO 2013153553 | Oct 2013 | WO |
| WO 2013175480 | Nov 2013 | WO |
| WO 2014022739 | Feb 2014 | WO |
| WO 2014106837 | Jul 2014 | WO |
| WO 2014106838 | Jul 2014 | WO |
| WO 2014151255 | Sep 2014 | WO |
| WO 2014164761 | Oct 2014 | WO |
| WO 2014164797 | Oct 2014 | WO |
| WO 2015010026 | Jan 2015 | WO |
| WO 2015200539 | Dec 2015 | WO |
| Entry |
|---|
| Brugière, Norbert, et al. “Glutamine synthetase in the phloem plays a major role in controlling proline production.” the Plant Cell 11.10 (1999): 1995-2011. (Year: 1999). |
| Riggins, Chance W., et al. “Characterization of de novo transcriptome for waterhemp (Amaranthus tuberculatus) using GS-FLX 454 pyrosequencing and its application for studies of herbicide target-site genes.” Pest management science 66.10 (2010): 1042-1052. (Year: 2010). |
| Tank mixing benefit, NCSU, 2004, published online at http://www.ncagr.gov/agronomi/pdffiles/Tank_Mixing.pdf (Year: 2004). |
| Wang, C. J., and Z. Q. Liu. “Foliar uptake of pesticides—present status and future challenge.” Pesticide Biochemistry and Physiology 87.1 (2007): 1-8. (Year: 2007). |
| Brugiére, Norbert, et al. “Glutamine synthetase in the phloem plays a major role in controlling proline production.” The Plant Cell 11.10 (1999): 1995-2011. (Year: 1999). |
| Declaration of Jerzy Zabkiewicz executed Nov. 28, 2017, as filed by Opponent in Australian Patent Application No. 2014262189, pp. 1-73, submitted in IDS on Aug. 7, 2019. (Year: 2017). |
| Ascencio-Ibanez et al., “DNA abrasion onto plants is an effective method for geminivirus infection and virus-induced gene silencing,” Journal of Virological Methods, 142:198-203 (2007). |
| Bachman et al., “Characterization of the spectrum of insecticidal activity of a double-stranded RNA with targeted activity against Western Corn Rootworm (Diabrotica virgifera virgifera LeConte),” Transgenic Res., pp. 1-16 (2013). |
| Bedell et al., “Sorghum Genome Sequencing by Methylation Filtration,” PLOS Biology, 3(1):E13/104-115 (2005). |
| Burgos et al., “Review: Confirmation of Resistance to Herbicides and Evaluation of Resistance Levels,” Weed Science, 61 (1):4-20 (2013). |
| Chen et al., “Exploring MicroRNA-Like Small RNAs in the Filamentous Fungus Fusarium oxysporum,” PLOS One, 9(8):e104956:1-10 (2014). |
| Constan et al., “An outer envelope membrane component of the plastid protein import apparatus plays an essential role in Arabidopsis,” the Plant Journal, 38:93-106 (2004). |
| Di Stilio et al., “Virus-Induced Gene Silencing as a Tool for Comparative Functional Studies in Thalictrum,” PLoS One, 5(8):e12064 (2010). |
| Eamens et al., “RNA Silencing in Plants: Yesterday, Today, and Tomorrow,” Plant Physiology, 147(2):456-468 (2008). |
| Fassler, BLAST Glossary, National Center for Biotechnology Information (2011). |
| Fernandez et al., “Uptake of Hydrophilic Solutes Through Plant Leaves: Current State of Knowledge and Perspectives of Foliar Fertilization,” Critical Reviews in Plant Sciences, 28:36-38 (2009). |
| Friedberg, “Automated protein function prediction—the genomic challenge,” Briefings in Bioinformatics, 7(3):225-242 (2006). |
| Funke et al., “Molecular basis for herbicide resistance in Roundup Ready crops,” PNAS, 103:13010-13015 (2006). |
| Gaskin et al., “Novel organosillicone adjuvants to reduce agrochemical spray volumes on row crops,” New Zealand Plant Protection, 53:350-354 (2000). |
| GenBank Accession No. EF143582 (2007). |
| Hagio, “Chapter 25: Direct Gene Transfer into Plant Mature Seeds via Electroporation After Vacuum Treatment,” Electroporation and Sonoporation in Developmental Biology, p. 285-293 (2009). |
| Hess, “Surfactants and Additives,” 1999 Proceedings of the California Weed Science Society, 51:156-172 (1999). |
| Huang et al., “In Vivo Analyses of the Roles of Essential Omp85-Related Proteins in the Chloroplast Outer Envelope Membrane,” Plant Physiol., 157:147-159 (2011). |
| Ivanova et al., “Members of the Toc159 Import Receptor Family Represent Distinct Pathways for Protein Targeting to Plastids,” Molecular Biology of the Cell, 15:3379-3392 (2004). |
| Jacque et al., “Modulation of HIV-1 replication by RNA interference,” Nature, 418, 435-438 (2002). |
| Jang et al., “Resistance to herbicides caused by single amino acid mutations in acetyl-CoA carboxylase in resistant populations of grassy weeds,” New Phytologist, 197(4):1110-1116 (2013). |
| Kikkert et al., “Stable Transformation of Plant Cells by Particle Bombardment/Biolistics,” Methods in Molecular Biology, 286:61-78 (2005). |
| Li et al., “A Simplified Seed Transformation Method for Obtaining Transgenic Brassica napus Plants,” Agricultural Sciences in China, 8(6):658-663 (2009). |
| Liu et al, “The Helicase and RNaseIIIa Domains of Arabidopsis Dicer-Like1 Modulate Catalytic Parameters during MicroRNA Biogenesis,” Plant Physiology, 159:748-758 (2012). |
| McGinnis, “RNAi for functional genomics in plants,” Brief Funct Genomics, 9(2):111-7 (2010). |
| Office Action dated Aug. 1, 2017, in European Patent Application No. 12 830 932.5. |
| Office Action dated Aug. 14, 2017, in Israeli Patent Application No. 235878. |
| Office Action dated Aug. 22, 2017, in Korean Patent Application No. 10-2012-7023415. |
| Office Action dated Aug. 3, 2017, in Chinese Patent Application No. 201480014392.5 (with English translation). |
| Office Action dated Aug. 3, 2017, in European Patent Application No. 12 831 684.1. |
| Office Action dated Aug. 8, 2017, in Chilean Patent Application No. 201501874. |
| Office Action dated Dec. 13, 2016, in Ukrainian Patent Application No. a 2014 03843. |
| Office Action dated Dec. 14, 2016, in Ukrainian Patent Application No. a 2014 03850. |
| Office Action dated Dec. 15, 2016, in Ukrainian Patent Application No. a 2014 03845. |
| Office Action dated Dec. 15, 2016, in Ukrainian Patent Application No. a 2014 03849. |
| Office Action dated Dec. 15, 2016, in Ukrainian Patent Application No. a 2014 03852. |
| Office Action dated Dec. 27, 2016, in Ukrainian Patent Application No. a 2012 11548. |
| Office Action dated Jul. 11, 2017, in Mexican Patent Application No. MX/a/2015/013118 (with English translation). |
| Office Action dated Jul. 3, 2017, in Mexican Patent Application No. MX/a/2015/012632 (with English translation). |
| Office Action dated Jul. 6, 2017, in Mexican Patent Application No. MX/a/2015/013103 (with English translation). |
| Office Action dated Mar. 16, 2017, in Chinese Patent Application No. 201280054819.5. |
| Office Action dated May 3, 2016, in Chilean Patent Application No. 201601057. |
| Office Action dated Nov. 15, 2016, in Mexican Patent Application No. MX/a/2014/003068 (with English translation). |
| Office Action dated Sep. 6, 2017, in Chinese Patent Application No. 2014800154012 (with English translation). |
| Patent Examination Report No. 1 dated Jun. 8, 2017, in Australian Patent Application No. 2012308686. |
| Powles et al., “Evolution in Action: Plants Resistant to Herbicides,” Annual Review of Plant Biology, 61(1):317-347 (2010). |
| Rakoczy-Trojanowska, “Alternative Methods of Plant Transformation—a short review,” Cellular & Molecular Biology Letters, 7:849-858 (2002). |
| Regalado, “The Next Great GMO Debate,” MIT Technology Review, pp. 1-19 (2015). |
| Richardson et al., “Targeting and assembly of components of the TOC protein import complex at the chloroplast outer envelope membrane,” Frontiers in Plant Science, 5:1-14 (2014). |
| Search Report dated Jul. 24, 2017, in Chinese Patent Application No. 201480014392.5 (with English translation). |
| Small, “RNAi for revealing and engineering plant gene functions,” Current Opinion in Biotechnology, 18:148-153 (2007). |
| Statement of Grounds and Particulars dated Sep. 1, 2017, in Australian Patent No. 2014262189. |
| Stevens, “Formulation of Sprays to Improve the Efficacy of Foliar Fertilisers,” New Zealand Journal of Forestry Science, 24(1):27-34 (1994). |
| Stevens, “Organosilicone Surfactants as Adjuvants for Agrochemicals,” New Zealand Journal of Forestry Science, 24:27-34 (1994). |
| Trucco et al., “Amaranthus hybridus can be pollinated frequently by A. tuberculatus under filed conditions,” Heredity, 94:64-70 (2005). |
| Voinnet, “Origin, Biogenesis, and Activity of Plant MicroRNAs,” Cell, 136:669-687 (2009). |
| Wild Carrot, Noxious Weed Control Board (NWCB) of Washington State (2010). |
| Wool et al., “Structure and evolution of mammalian ribosomal proteins,” Biochem. Cell Biol., 73:933-947 (1995). |
| Written Opinion dated Mar. 6, 2017, in Singaporean Patent Application No. 2012061529. |
| Zhang, “Artificial trans-acting small interfering RNA: a tool for plant biology study and crop improvements,” Planta, 239:1139-1146 (2014). |
| Zhang, Chapter 10: New Characteristics of Pesticide Research & Development, p. 209 (2010). |
| Zhong et al., “A forward genetic screen to explore chloroplast protein import in vivo identifies Moco sulfurase, pivotal for ABA and IAA biosynthesis and purine turnover,” The Plant Journal, 63:44-59 (2010). |
| Agricultural Chemical Usage 2006 Vegetables Summary, Agricultural Statistics Board, NASS, USDA, pp. 1-372 (2007). |
| Agrios, Plant Pathology (Second Edition), 2:466-470 (1978). |
| Alarcón-Reverte et al., “Resistance to ACCase-inhibiting herbicides in the weed Lolluin multiflorum,” Comm. Appl. Biol. Sci., 73(4):899-902 (2008). |
| Al-Kaff el al., “Plants rendered herbicide-susceptible by cauliflower mosaic virus-elicited suppression of a 35S promoter-regulated transgene,” Nature Biotechnology, 18:995-999 (2000). |
| Amarzguioui et al., “An algorithm for selection of functional siRNA sequences,” Biochemical and Biophysical Research Communications, 316:1050-1058 (2004). |
| Ambrus et al., “The Diverse Roles of RNA Helicases in RNAi,” Cell Cycle, 8(21):3500-3505 (2009). |
| An et al., “Transient RNAi Induction against Endogenous Genes in Arabidopsis Protoplasts Using in Vitro-Prepared Double-Stranded RNA,” Biosci Biotechnol Biochem, 69(2):415-418 (2005). |
| Andersson et al., “A novel selection system for potato transformation using a mutated AHAS gene,” Plant Cell Reports, 22(4):261-267 (2003). |
| Anonymous, “A handbook for high-level expression and purification of 6xHis-tagged proteins,” The QiaExpressionist, (2003). |
| Anonymous, “Agronomy Facts 37: Adjuvants for enhancing herbicide performance,” n.p., 1-8, (Jan. 26, 2000), Web, (Jan. 21, 2014). |
| Anonymous, “Devgen, The mini-Monsanto,” KBC Securities (2006). |
| Anonymous, “Do Monsanto have the next big thing?,” Austalian Herbicide Resistance Initiative (AHRI), (Apr. 23, 2013) Web. (Jan. 19, 2015). |
| Aoki et al., “In Vivo Transfer Efficiency of Antisense Oligonucleotides into the Myocardium Using HVJ-Liposome Method,” Biochem Biophys Res Common, 231:540-545 (1997). |
| Arpaia et al., “Production of transgenic eggplant (Solanum melongena L.) resistant to Colorado Potato Beetle (Leptinotarsa decemlineata Say),” (1997) Theor. Appl. Genet., 95:329-334 (1997). |
| Artymovich, “Using RNA interference to increase crop yield and decrease pest damage,” MMG 445 Basic Biotech., 5(1):7-12 (2009). |
| Australian Patent Examination report No. 1 dated Nov. 11, 2013, in Australian Application No. 2011224570. |
| Axtell et al., “A Two-Hit Trigger for siRNA Biogenesis in Plants,” Cell, 127:565-577 (2006). |
| Baerson et al., “Glyphosate-Resistant Goosegrass. Identification of a Mutation in the Target Enzyme 5-Enolpyruvylshikimate-3-Phosphate Synthase,” Plant Physiol., 129(3):1265-1275 (2002). |
| Bai et al., “Naturally Occurring Broad-Spectrum Powdery Mildew Resistance in a Central American Tomato Accession is Caused by Loss of Mlo Function,” MPMI, 21(1):30-39 (2008). |
| Balibrea et al., “Extracellular Invertase is an Essential Component of Cytokinin-Mediated Delay of Senescence,” The Plant Cell, 16(5):1276-1287 (2004). |
| Bannerjee et al., “Efficient production of transgenic potato (S. tuberosum L. ssp. andigena) plants via Agrobacterium tumefactens-mediated transformation,” Plant Sci., 170:732 738 (2006). |
| Bart et al., “A novel system for gene silencing using siRNAs in rice leaf and stem-derived protoplasts,” Plant Methods, 2(13):1-9 (2006). |
| Basu et al., “Weed genomics: new tools to understand weed biology,” Trends in Plant Science, 9(8):391-398 (2004). |
| Baulcombe, “RNA silencing and heritable epigenetic effects in tomato and Arabidopsis,” Abstract 13th Annual Fall Symposium, Plant Genomes to Phenomes, Donald Danforth Plant Science Center, 28-30 (2011). |
| Bayer et al., “Programmable ligand-controlled riboregulators of eukaryotic gene expression,” Nature Biotechnol., 23(3):337-343 (2005). |
| Beal, et al., “Second Structural Motif for Recognition of DNA by Oligonucleotide-Directed Triple-Helix Formation,” Science, 251:1360-1363 (1992). |
| Becker et al., “Fertile transgenic wheat from microprojectile bombardment of scutellar tissue,” The Plant Journal, 5(2):299-307 (1994). |
| Bhargava et al., “Long double-stranded RNA-mediated RNA interference as a tool to achieve site-specific silencing of hypothalamic neuropeptides,” Brain Research Protocols, 13:115-125 (2004). |
| Boletta et al., “High Efficient Non-Viral Gene Delivery to the Rat Kidney by Novel Polycationic Vectors,” J. Am Soc. Nephrol., 7:1728 (1996). |
| Bolognesi et al., “Characterizing the Mechanism of Action of Double-Stranded RNA Activity against Western Corn Rootworm(Diabrotica virgifera virgifera LeConte),” PLoS One 7(10):e47534 (2012). |
| Bolter et al., “A chloroplastic inner envelope membrane protease is essential for plant development,” FEBS Letters, 580:789-794 (2006). |
| Bourgeois et al., “Field and producer survey of ACCase resistant wild oat in Manitoba,” Canadian Journal of Plant Science, 709-715 (1997). |
| Breaker et al., “A DNA enzyme with Mg2+-dependent RNA phosphoesterase activity,” Chemistry and Biology, 2:655-660 (1995). |
| Brodersen et al., “The diversity of RNA silencing pathways in plants,” Trends in Genetics, 22(5):268-280 (2006). |
| Brugiere et al., “Glutamine Synthetase in the Phloem Plays a Major Role in Controlling Proline Production,” The Plant Cell, 11:1995-2011 (1999). |
| Busch el al., “RNAi for discovery of novel crop protection products,” Pflanzenschutz-Nachrichten Bayer, 58(1):34-50 (2005). |
| Busi et al., “Gene flow increases the initial frequency of herbicide resistance alleles in unselectedpopulations,” Agriculture, Ecosystems and Environments, Elsevier, Amsterdam, NL, 142(3):403-409 (2011). |
| Butler et al., “Priming and re-drying improve the survival of mature seeds of Digitalis purpurea during storage,” Annals of Botany, 103:1261-1270 (2009). |
| Bytebier et al., “T-DNA organization in tumor cultures and transgenic plants of the monocotyledon Asparagus officinalis,” Proc. Natl. Acad. Sci. U.S.A., 84:5345-5349 (1987). |
| Campbell et al., “Gene-knockdown in the honey bee mite Varroa destructor by a non-invasive approach: studies on a glutathione S-transferase,” Parasites & Vectors, 3(1):73, pp. 1-10 (2010). |
| Chabannes et al., “In situ analysis of lignins in transgenic tobacco reveals a differential impact of individual transformations on the spatial patterns of lignin deposition at the cellular and subcellular levels,” The Plant Journal, 28(3):271-282 (2001). |
| Chabbouh et al., “Cucumber mosaic virus in artichoke,” FAO Plant Protection Bulletin, 38:52-53 (1990). |
| Chakravarty et al., “Genetic Transformation in Potato: Approaches and Strategies,” Amer J Potato Res, 84:301 311 (2007). |
| Chang et al., “Cellular Internalization of Fluorescent Proteins via Arginine-rich Intracellular Delivery Peptide in Plant Cells,” Plant Cell Physiol., 46(3):482-488 (2005). |
| Chee et al., “Transformation of Soybean (Glycine max) by Infecting Germinating Seeds with Agrobacterium tumefaciens,” Plant Physiol., 91:1212-1218 (1989). |
| Chen et al., “Tn Vivo Analysis of the Role of atTic20 in Protein import into Chloroplasts,” The Plant Cell, 14:641-654 (2002). |
| Chen et al., “Transfection and Expression of Plasmid DNA in Plant Cells by an Arginine-Rich Intracellular Delivery Peptide without Protoplast Preparation,” FEBS Letters 581, pp. 1891-1897 (2007). |
| Cheng et al., “Production of fertile transgenic peanut (Arachis hypogaea L.) plants using Agrobacterium tumefaciens,” Plant Cell Reports, 15:653-657 (1996). |
| Chi et al., “The Function of RH22, a DEAD RNA Helicase, in the Biogenesis of the 50S Ribosomal Subunits of Arabidopsis Chloroplasts,” Plant Physiology, 158:693-707 (2012). |
| Chupp et al., “Chapter 8: White Rust,” Vegetable Diseases and Their Control, the Ronald Press Company, New York, pp. 267-269 (1960). |
| Clough et al., “Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana,” The Plant Journal, 16(6):735-743 (1998). |
| CN101914540 Patent Diclosure, “Introduction of RNA into plant by interference,” (2010). |
| Colbourne et al.,“The Ecoresponsive Genome of Daphnia pulex,” Science, 331(6017):555-561 (2011). |
| Colombian Office Action dated Aug. 2, 2013 in Application No. 12 152898. |
| Colombian Office Action dated Feb. 21, 2014 in Application No. 12 152898. |
| Colliver et al., “Differential modification of flavonoid and isoflavonoid biosynthesis with an antisense chalcone synthase construct in transgenic Lotus corniculatus,” Plant Molecular Biology, 35:509-522 (1997). |
| Communication pursuant to Article 94(3) EPC dated Jan. 14, 2016, in European Patent Application No. 12 832 415.9. |
| Communication pursuant to Article 94(3) EPC dated Jun. 26, 2015, in European Patent Application No. 11 753 916.3. |
| Communication pursuant to Article 94(3) EPC dated Mar. 18, 2016, in European Patent Application No. 12 832 160.1. |
| Communication pursuant to Article 94(3) EPC dated Mar. 24, 2016, in European Patent Application No. 12 831 684.1. |
| Communication pursuant to Article 94(3) EPC dated Mar. 4, 2016, in European Patent Application No. 12 830 932.5. |
| Communication pursuant to Article 94(3) EPC dated Mar. 9, 2016, in European Patent Application No. 12 831 166.9. |
| Communication pursuant to Article 94(3) EPC dated Oct. 23, 2015, in European Patent Application No. 12 831 945.6. |
| Concise Descriptions of Relevance filed by a third party on Nov. 29, 2012, in U.S. Appl. No. 13/042,856. |
| Cooney et al., “Site-Specific Oligonucleotide Binding Represses Transcription of the Human c-myc Gene in Vitro,” Science ,241:456-459 (1988). |
| COST Action FA0806 progress report “Plant virus control employing RNA-based vaccines: A novel non-transgcnic strategy” (2010). |
| Coticchia et al., “Calmodulin modulates Akt activity in human breast cancer cell lines,” Breast Cancer Res. Treat, 115:545-560 (2009). |
| Dalakouras et al., “Induction of Silencing in Plants by High-Pressure Spraying of hi vitro-Synthesized Small RNAs,” Frontiers in Plant Science, 7(1327):1-5 (2016). |
| Dalmay et al., “An RNA-Depenedent RNA Polymerase Gene in Arabidopsis is Required for Posttranscriptional Gene Silencing Mediated by a Transgene but not by a Virus,” Cell, 101:543-553 (2000). |
| Davidson et al., “Engineering regulatory RNAs,” Trends in Biotechnology, 23(3):109-112 (2005). |
| Dawson et al., “cDNA cloning of the complete genome of tobacco mosaic virus and production of infectious transcripts,” Proc. Natl. Acad. Sci. USA, 83:1832-1836 (1986). |
| De Block, et al. “Engineering herbicide resistance in plants by expression of a detoxifying enzyme,” EMBO J. 6(9):2513-2519 (1987). |
| De Framond, “Mini-Ti: A New Vector Strategy for Plant Genetic Engineering,” Nature Biotechnology, 1:262-269 (1983). |
| Della-Cioppa et al., “Import of a precursor protein into chloroplasts is inhibited by the herbicide glyphosate,” The EMBO Journal, 7(5):1299-1305 (1988). |
| Desai et al., “Reduction in deformed wing virus infection in larval and adult honey bees (Apis mellifera L.) by double-stranded RNA ingestion,” Insect Molecular Biology, 21(4):446-455 (2012). |
| Diallo el al., “Long Endogenous dsRNAs Can Induce Complete Gene Silencing in Mammalian Cells and Primary Cultures,” Oligonucleotides, 13:381-392 (2003). |
| Dietemann et al.,“Varroa destructor: research avenues towards sustainable control,” Journal of Apicultural Research, 51(1):125-132 (2012). |
| Du et al., “A systematic analysis of the silencing effects of an active siRNA at all single-nucleotide mismatched target sites,” Nucleic Acids Research, 33(5):1671-1677 (2005). |
| Dunoyer et al., “Small RNA Duplexes Function as Mobile Silencing Signals Between Plant Cells,” Science, 328:912-916 (2010). |
| Ellington et al., “In vitro selection of RNA molecules that bind specific ligands,” Nature, 346:818-822 (1990). |
| Emery et al., “Radial Patterning of Arabidopsis Shoots by Class III HD-ZIP and KANADI Genes,” Current Biology, 13:1768-1774 (2003). |
| European Cooperation in the field of Scientific and Technical Research—Memorandum of Understanding for COST Action FA0806 (2008). |
| Extended European Search Report dated Feb. 2, 2015, in European Patent Application No. 12 830 932.5. |
| Extended European Search Report dated Feb. 27, 2015, in European Patent Application No. 12 832 160.1. |
| Extended European Search Report dated Feb. 3, 2015, in European Patent Application No. 12 831 945.6. |
| Extended European Search Report dated Jan. 20, 2016, in European Patent Application No. 13 794 339.5. |
| Extended European Search Report dated Jan. 21, 2015, in European Patent Application No. 12 832 415.9. |
| Extended European Search Report dated Jan. 29, 2015, in European Patent Application No. 12 831 567.8. |
| Extended European Search Report dated Jun. 29, 2015, in European Patent Application No. 12 831 494.5. |
| Extended European Search Report dated Mar. 17, 2015, in European Patent Application No. 12 831 684.1. |
| Extended European Search Report dated Mar. 3, 2015, in European Patent Application No. 12 831 166.9. |
| Extended European Search Report dated Oct. 8, 2013, in European Patent Application No. 11753916.3. |
| Extended European Search Report dated Sep. 29, 2016, in European Patent Application No. 14778840.0. |
| Farooq et al.. “Rice seed priming,” IPRN, 30(2):45-48 (2005). |
| Feuillet et al., “Crop genome sequencing: lessons and rationales,” Trends Plant Sci., 16:77-88 (2011). |
| Final Office Action dated Apr. 7, 2016, in U.S. Appl. No. 13/619,980. |
| Final Office Action dated Dec. 17, 2015, in U.S. Appl. No. 14/335,135. |
| Final Office Action dated Feb. 17, 2016, in U.S. Appl. No. 13/612,929. |
| Final Office Action dated Feb. 4, 2016, in U.S. Appl. No. 13/612,936. |
| Final Office Action dated Jun. 30, 2016, in U.S. Appl. No. 13/901,326. |
| Final Office Action dated Mar. 2, 2016, in U.S. Appl. No. 13/612,995. |
| Final Office Action dated Mar. 21, 2016, in U.S. Appl. No. 13/612,925. |
| Final Office Action dated May 26, 2016, in U.S. Appl. No. 14/532,596. |
| Final Office Action dated Sep. 9, 2016, in U.S. Appl. No. 13/612,954. |
| Final Office Action dated Nov. 10, 2015, in U.S. Appl. No. 13/612,985. |
| Final Office Action dated Nov. 19, 2015, in U.S. Appl. No. 13/612,941. |
| Final Office Action dated Nov. 10, 2016, in U.S. Appl. No. 13/583,302. |
| Final Office Action dated Nov. 30, 2015, in U.S. Appl. No. 13/612,948. |
| Final Office Action dated Sep. 9, 2016, in U.S. Appl. No. 14/608,951. |
| Final Office Action dated Sep. 9, 2016, in U.S. Appl. No. 14/603,347. |
| Final Office Action dated Nov. 7, 2013, in U.S. Appl. No. 13/042,856. |
| Final Office Action dated Oct. 20, 2016, in U.S. Appl. No. 14/480,199. |
| Final Office Action dated Oct. 22, 2015, in U.S. Appl. No. 14/608,951. |
| Fire et al., “Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans,” Nature, 391:806-811 (1998). |
| First Examination Report dated Apr. 23, 2013, in New Zealand Patent Application No. 601784. |
| First Examination Report dated Jul. 28, 2014, in New Zealand Patent Application No. 627060. |
| First Office Action dated Aug. 31, 2015, in Chinese Patent Application No. 201280053985.3. |
| First Office Action dated Feb. 2, 2016, in Chinese Patent Application No. 201380039346.6. |
| First Office Action dated Jul. 7, 2015, in Chinese Patent Application No. 201280054820.8. |
| First Office Action dated Mar. 12, 2015, in Chinese Patent Application No. 201280053984.9. |
| First Office Action dated Mar. 2, 2015, in Chinese Patent Application No. 201280054819.5. |
| First Office Action dated May 27, 2015, in Chinese Patent Application No. 201280054179.8. |
| First Office Action dated Sep. 9, 2015, in Chinese Patent Application No. 201280055409.2. |
| Fraley et al., “Liposome-mediated delivery of tobacco mosaic virus RNA into tobacco protoplasts: A sensitive assay for monitoring liposome-protoplast interactions,” Proc Natl Acad Sci U S A., 79(6):1859-1863 (1982). |
| Fukuhara et al., “Enigmatic Double-Stranded RNA in Japonica Rice,” Plant Molecular Biology, 21:1121-1130 (1993). |
| Fukuhara el al., “The Unusual Structure of a Novel Rna Replicon in Rice,” The Journal of Biological Chemistry, 270(30):18147-18149 (1995). |
| Fukuhara et al., “The wide distribution of endornaviruses, large double-stranded RNA replicons with plasmid-like properties,” Archives of Virology, 151:995-1002 (2006). |
| Fukunaga et al., “dsRNA with 5′ overhangs v contributes to endogenous and antiviral RNA silencing pathways in plants,” The EMBO Journal, 28(5):545-555 (2009). |
| Further Examination Report dated May 16, 2014, in New Zealand Patent Application No. 601784. |
| Gaines et al., “Gene amplification confers glyphosate resistance in Amaranthus Palmeri,” Proc. Natl. Acad. Sci. USA, 107(3):1029-1034 (2010). |
| Gallie et al., “Identification of the motifs within the tobacco mosaic virus 5′-leader responsible for enhancing translation,” Nucleic Acids Res., 20(17):4631-4638 (1992). |
| Gan et al., “Bacterially expressed dsRNA protects maize against SCMV i nfection,” Plant Cell Rep, 11:1261-1268 (2010). |
| Gan et al., “Inhibition of Leaf Senescence by Autoregulated Production of Cytokinin,” Science, 270:1986-1988 (1995). |
| Gao et al., “Down-regulation of acetolactate synthase compromises 01-1-mediated resistance to powdery mildew in tomato,” BMC Plant Biology, 14 (2014). |
| Gao et al., “Nonviral Methods for siRNA Delivery,” Molecular Pharmaceutics, 6(3):651-658 (2008). |
| Garbian et al., “Bidirectional Transfer of RNAi between Honey Bee and Varroa destructor: Varroa Gene Silencing Reduces Varroa Population,” 8(12):1-9:e1003035 (2012). |
| Ge et al., “Rapid vacuolar sequestration: the horseweed glyphosate resistance mechanism,” Pest Management Sci., 66:345-348 (2010). |
| GenBank Accession No. AY545657.1 (2004). |
| GenBank Accession No. CB377464, “CmaE1_37_J02J3 Cowpea weevil larvae Lambda Zap Express Library Callosobruchus maculatus cDNA, mRNA sequence,” (2007). |
| GenBank Accession No. DY640489, “PU2_plate27_F03 PU2 Prunus persica cDNA similar to expressed mRNA inferred from Prunus persica hypothetical domain/motif cont aining IPR011005:Dihydropteroate synthase-like, MRNA sequence” (2006). |
| GenBank Accession No. EU024568, “Amaranthus hypochondriacus acetolactate synthase (ALS) gene” (2007). |
| GenBank Accession No. EW765249, “ST020010B10C12 Normalized and subtracted western corn rootworm female head cDNA library Diabrotica virgifera virgifera cDNA clone STO20010B10C12 5-, mRNA sequence,” (2007). |
| GenBank Accession No. EW771198, “ST020010B10C12 Normalized and subtracted western corn rootworm female head cDNA library Diabrotica virgifera virgifera cDNA clone STO20010B10C12 5-, mRNA sequence,” (2007). |
| GenBank Accession No. Fe,348695, “CBIB7954.fwd CBIB_Daphnia_pulex_Chosen_One_Library_2 Daphnia pulex cDNA clone CB1B7954 5′, mRNA sequence” (2011). |
| GenBank Accession No. FJ972198, “Solanum lycopersicum cultivar Ailsa Craig dihydropterin pyrophosphokinase-dihydropteroate synthase (HPPK-DHPS) gene, complete cds” (2010). |
| GenBank Accession No. GI:186478573 (2014). |
| GenBank Accession No. GU120406, “Chrysomela tremulae ribosomal protein L7 (RpL7) mRNA, complete cds” (2009). |
| GenBank Accession No. HD315444, “Sequence 192160 from Patent EP2213738” (2010). |
| GenBank Accession No. Q4GXM3_BIPLU, “Ribosomal protein L7e” (2006). |
| GenBank Accession No. U87257.1, “Daucus carota 4-hydroxyphenylpyruvate dioxygenase mRNA, complete cds” (1997). |
| GenBank Accession No. XM_014456745.1, Predicted: Myotis lucifugus ribonucleoprotein, PTB-binding 2 (RAVER2), transcript variant X3, mRNA,: (2015). |
| GenBank Accession No. Y08611.1, “P.sativum mRNA for dihydropterin pyrophosphokinase/dihydropteroate synthase.” (2006). |
| GenEmbl Accession No. FJ861243 (2010). |
| Gong et al., “Silencing of Rieske iron-sulfur protein using chemically synthesised siRNA as a potential biopesticide against Plutella xylostella,” Pest Manag Sci, 67:514-520 (2011). |
| Gossamer Threads, Compendium of Herbicide Adjuvants: Organo-Silicone Surfactant, p. 1-4 (1998). |
| Gressel et al., “A strategy to provide long-term control of weedy rice while mitigating herbicide resistance transgene flow, and its potential use for other crops with related weeds,” Pest Hanag Sci, 65(7):723-731 (2009). |
| Gudkov, “Minireview: The L7/L12 ribosomal domain of the ribosome: structural and functional studies,” FEBS Letters, 407:253-256 (1997). |
| Gutensohn et al., “Functional analysis of the two Arabidopsis homologues of Toc34, a component of the chloroplast protein import apparatus,” The Plant Journal, 23(6):771-783 (2000). |
| Haigh, “The Priming of Seeds: Investigation into a method of priming large quantities of seeds using salt solutions,” Thesis submitted to Macquarie University (1983). |
| Hajirezaei et al., “Impact of elevated cytosolic and apoplastic invertase activity on carbon metabolism during potato tuber development,” Journal of Experimental Botany, 51:439-445 (2000). |
| Hamilton et al., “Guidelines for the Identification and Characterization of Plant Viruses,” J. gen. Virol., 54:223-241 (1981). |
| Hamilton et al., “Two classes of short interfering RNA in RNA silencing,” EMBO J., 21(17):4671-4679 (2002). |
| Han et al., “Molecular Basis for the Recognition of Primary microRNAs by the Drosha-DGCR8 Complex,” Cell, 125(5):887-901 (2006). |
| Hannon, “RNA interference,” Nature,481:244-251 (2002). |
| Hardegree, “Drying and storage effects on germination of primed grass seeds,” Journal of Range Management, 47(3):196-199 (1994). |
| Harrison et al., “Does Lowering Glutamine Synthetase Activity in Nodules Modigy Nitrogen Metabolism and Growth of Lotus japonicus?,” Plant Physiology, 133:253-262 (2003). |
| Heffer et al., “Rapid isolation of gene homologs across taxa: Efficient identification and isolation of gene orthologs from non-model organism genomes, a technical report,” EvoDevo Journal, 2(7):1-5 (2011). |
| Herman et al., “A three-component dicamba O-demethylase from Pseudomonas maltophilia, strain DI-6: gene isolation, characterization, and heterologous expression,” J. Biol. Chem., 280: 24759-24767 (2005). |
| Hewezi et al., “Local infiltration of high- and low-molecular-weight RNA from silenced sunflower (Helianthus annuus L.) plants triggers post-transcriptional gene silencing in non-silenced plants,” Plant Biotechnology Journal, 3:81-89 (2005). |
| Hidayat et al., “Enhanced Metabolism of Fluazifop Acid in a Biotype of Digitaria sanguinalis Resistant to the Herbicide Fluazifop-P-Butyl,” Pesticide Biochem. Physiol., 57:137-146 (1997). |
| Himber et al., “Transitivity-dependant and -independent cell-to-cell movement of RNA silencing,” The EMBO Journal, 22(17):4523-4533 (2003). |
| Hirschberg et al., “Molecular Basis of Herbicide Resistance in Amaranthus hybridus,” Science, 222:1346-1349 (1983). |
| Hoekema et al., “A binary plant vector strategy based on separation of vir- and T-region of the Agrobacterium tumefaciens Ti-plasmid,” Nature, 303:179-180 (1983). |
| Hofgen et al., “Repression of Acetolactate Synthase Activity through Antisense Inhibition: Molecular and Biochemical Analysis of Transgenic Potato (Solarium tuberosum L. cv Desiree) Plants,” Plant Physiol., 107(2):469-477 (1995). |
| Holtra et al., “Assessment of the Physiological Condition of Salvinia Natans L. Exposed to Copper(II) Ions,” Environ. Protect. Eng., 41:147-158 (2015). |
| Hsieh et al., “A library of siRNA duplexes targeting the phosphoinositide 3-kinase pathway: determinants of gene silencing for use in cell-based screens,” Nucleic Acids Res., 32(3):893-901 (2004). |
| Huesken et al., “Design of a genome-wide siRNA library using an artificial neural network,” Nature Biotechnology, 23(8): 995-1001 (2005). |
| Hunter et al., “RNA Interference Strategy to suppress Psyllids & Leafhoppers,” International Plant and Animal Genome XIX, 15-19 (2011). |
| Ichihara et al., “Thermodynamic instability of siRNA duplex is a prerequisite for dependable prediction of siRNA activities,” Nucleic Acids Res., 35(18):e123 (2007). |
| International Preliminary Report on Patentability (Chapter II) dated Jul. 24, 2015, in International Application No. PCT/US2014/047204. |
| International Preliminary Report on Patentability dated Sep. 11, 2012, in International Application No. PCT/US2011/027528. |
| International Preliminary Report on Patentability dated Sep. 11, 2014, in International Application No. PCT/IL2013/050447. |
| International Rice Genome Sequencing Project, the map-based sequence of the rice genome, Nature, 436(11):793-800 (2005). |
| International Search Report and the Written Opinion dated Feb. 25, 2013, in International Application No. PCT/US2012/054883. |
| International Search Report and the Written Opinion dated Feb. 27, 2013, in International Application No. PCT/US2012/054814. |
| International Search Report and the Written Opinion dated Feb. 27, 2013, in International Application No. PCT/US2012/054842. |
| International Search Report and the Written Opinion dated Feb. 27, 2013, in International Application No. PCT/US2012/054862. |
| International Search Report and the Written Opinion dated Feb. 27, 2013, in International Application No. PCT/US2012/054894. |
| International Search Report and the Written Opinion dated Feb. 27, 2013, in International Application No. PCT/US2012/054974. |
| International Search Report and the Written Opinion dated Feb. 27, 2013, in International Application No. PCT/US2012/054980. |
| International Search Report and the Written Opinion dated Jul. 15, 2014, in International Application No. PCT/US2014/025305. |
| International Search Report and the Written Opinion dated Jul. 22, 2014, in International Application No. PCT/IL2013/051083. |
| International Search Report and the Written Opinion dated Jul. 22, 2014, in International Application No. PCT/IL2013/051085. |
| International Search Report and the Written Opinion dated Jul. 24 2014, in International Application No. PCT/US2014/026036. |
| International Search Report and the Written Opinion dated May 10, 2011, in International Application No. PCT/US2011/027528. |
| International Search Report and the Written Opinion dated Oct. 1, 2013, in International Application No. PCT/IL2013/050447. |
| International Search Report and Written Opinion dated Aug. 25, 2014, in International Application No. PCT/US2014/023503. |
| International Search Report and Written Opinion dated Aug. 27, 2014, in International Application No. PCT/US2014/023409. |
| International Search Report and Written Opinion dated Feb. 23, 2015, in International Application No. PCT/US2014/063832. |
| International Search Report and Written Opinion dated Jul. 8, 2015, in International Application No. PCT/US2015/011408. |
| International Search Report and Written Opinion dated Mar. 26, 2015, in International Application No. PCT/US2014/069353. |
| International Search Report and Written Opinion dated May 26, 2016, in International Application No. PCT/US2016/014344. |
| International Search Report and Written Opinion dated Nov. 24, 2015, in International Application No. PCT/US2015/037522. |
| International Search Report and Written Opinion dated Nov. 27, 2015, in International Application No. PCT/US2015/037015. |
| International Search Report dated Mar. 12, 2013, in International Application No. PCT/US2012/054789. |
| Invitation to Pay Additional Fees dated May 6, 2014, in International Application No. PCT/IL2013/051083. |
| Invitation to Pay Additional Fees dated May 6, 2014, in International Application No. PCT/IL2013/051085. |
| Invitation to Pay Additional Fees dated Nov. 25, 2014, in International Application No. PCT/US2014/047204. |
| Invitation to Pay Additional Fees dated Sep. 8, 2015, in International Application No. PCT/US2015/037015. |
| Invitation to Pay Additional Fees dated Sep. 9, 2015, in International Application No. PCT/US2015/037522. |
| Isaacs et al., “Engineered riboregulators enable post-transcriptional control of gene expression,” Nature Biotechnology, 22(7):841-847 (2004). |
| Ji et al., “Regulation of small RNA stability: methylation and beyond,” Cell Research, 22:624-636 (2012). |
| Jin et al., “Posttranslational Elevation of Cell Wall Invertase Activity by Silencing its Inhibitor in Tomato Delays Leaf Senescence and Increases Seed Weight and Fruit Hexose Level,” The Plant Cell, 21:2072-2089 (2009). |
| Jofre-Garfias et al., “Agrobacterium-mediated transformation of Amaranthus hypochondriacus: light- and tissue-specific expression of a pea chlorophyll a/b-binding protein promoter,” Plant Cell Reports, 16:847-852 (1997). |
| Jones-Rhoades et al., “MicroRNAs and Their Regulatory Roles in Plants,” Annu. Rev. Plant Biol., 57:19-53 (2006). |
| Josse et al., “A DELLA in Disguise: SPATULA Restrains the Growth of the Developing Arabidopsis Seedling,” Plant Cell, 23:1337-1351 (2011). |
| Kaloumenos et al., “Identification of a Johnsongrass (Sorghum halepense) Biotype Resistant to ACCase-Inhibiting Herbicides in Northern Greece,” Weed Technol, 23:470-476 (2009). |
| Kam et al., “Nanotube Molecular Transporters: Internalization of Carbon Nanotube-Protein Conjugates into Mammalian Cells,” J. Am. Chem. Soc., 126(22):6850-6851 (2004). |
| Kambiranda et al., “Relationship Between Acid Invertase Activity and Sugar Content in Grape Species,” Journal of Food Biochemistry, 35:1646-1652 (2011). |
| Katoh et al., “Specific residues at every third position of siRNA shape its efficient RNAi activity,” Nucleic Acids Res., 35(4): e27 (2007). |
| Kertbundit et al.,“In vivo random β-glucuronidase gene fusions in Arabidopsis thaliana,” Proc. Natl. Acad. Sci. U S A., 88:5212-5216 (1991). |
| Khachigian, “DNAzymes: Cutting a path to a new class of therapeutics,” Curr Opin Mol Ther 4(2):119-121 (2002). |
| Khan et al., “Matriconditioning of Vegetable Seeds to Improve Stand Establishment in Early Field Plantings,” J. Amer. Soc. Hort. Sci., 117(1):41-47 (1992). |
| Khodakovskaya et al., “Carbon Nanotubes are Able to Penetrate Plant Seed Coat and Dramatically Affect Seed Germination and Plant Growth,” ACS Nano, 3(10):3221-3227 (2009). |
| Kim et al., “Optimization of Conditions for Transient Agrobacterium-Mediated Gene Expression Assays in Arabidopsis,” Plant Cell Reports, 28:1159-1167 (2009). |
| Kim el al., “Synthetic dsRNA Dicer substrates enhance RNAi potency and efficacy,” Nature Biotechnology, 23(2):222-226 (2005). |
| Kirkwood, “Herbicides and Plants,” Botanical Journal of Scotland, 46(3):447-462 (1993). |
| Kirkwood, “Use and Mode of Action of Adjuvants for Herbicides: A Review of some Current Work,” Pestic Sci., 38:93-102 (1993). |
| Klahre et al., “High molecular weight RNAs and small interfering RNAs induce systemic posttranscriptional gene silencing in plants,” Proc. Natl. Acad. Sci. USA, PNAS, 99(18):11981-11986 (2002). |
| Knudsen, “Promoter2.0: for the recognition of Poll promoter sequences,” Bioniformatics, 15(5):356-361 (1999). |
| Kronenwett et al., “Oligodeoxyribonucleotide Uptake in Primary Human Hematopoietic Cells is Enhanced by Cationic Lipids and Depends on the Hematopoietic Cell Subset,” Blood, 91(3):852-862 (1998). |
| Kumar et al., “Sequencing, De Novo Assembly and Annotation of the Colorado Potato Beetle, Leptinotarsa decemlineata,Transcriptome,” PLoS One, 9(1):e86012 (2014). |
| Kusaba et al., “Low glutelin content1: A Dominant Mutation That Suppresses the Glutelin Multigene Family via RNA Silencing ni Rice,” The Plant Cell, 15(6):1455-1467 (2003). |
| Kusaba, “RNA interference in crop plants,” Curr Opin Biotechnol, 15(2):139-143 (2004). |
| Lavigne et al., “Enhanced antisense inhibition of human immunodeficiency virus type 1 in cell cultures by DLS delivery system,” Biochem Biophys Res Common, 237:566-571 (1997). |
| Lee et al., “Aptamer Database,” Nucleic Acids Research, 32:D95-D100 (2004). |
| Lein et al., “Target-based discovery of novel herbicides,” Current Opinion in Plant Biology, 7:219-225 (2004). |
| Leopold et al., “Chapter 4: Moisture as a Regulator of Physiological Reaction in Seeds,” Seed Moisture, OSSA Special Publication No. 14, pp. 51-69 (1989). |
| Lermontova et al., “Reduced activity of plastid protoporphyrinogen oxidase causes attenuated photodynamic damage during high-light compared to low-light exposure,” The Plant Journal, 48(4):499-510 (2006). |
| Lesnik et al., “Prediction of rho-independent transcriptional terminators in Escherichia coli,” Nucleic Acids Research, 29(17):3583-3594 (2001). |
| Li et al., “Establishment of a highly efficient transformation system for pepper (Capsicum annuum L.),” Plant Cell Reports, 21: 785-788 (2003). |
| Li et al., “The FAST technique: a simplified Agrobacterium-based transformation method for transient gene expression analysis in seedlings of Arabidopsis and other plant species,” Plant Methods, 5(6):1-15 (2009). |
| Liu et al., “Carbon Nanotubes as Molecular Transporters for Walled Plant Cells,” Nano Letters, 9(3):1007-1010 (2009). |
| Liu et al., “Comparative study on the interaction of DNA with three different kinds of surfactants and the formation of multilayer films,” Bioelectrochemistry, 70:301-307 (2007). |
| Liu et al., “DNAzyme-mediated recovery of small recombinant RNAs from a 5S rRNA-derived chimera expressed in Escherichia coli,” BMC Biotechnology, 10:85 (2010). |
| Liu et al., “Identification and Application of a Rice Senescence-Associated Promoter,” Plant Physiology, 153:1239-1249 (2010). |
| Liu, “Influence of Sugars on the Foliar Uptake of Bentazone and Glyphosate,” New Zealand Plant Protection, 55:159-162 (2002). |
| Llave et al., “Endogenous and Silencing-Associated Small RNAs in Plants,” The Plant Cell, 14:1605-1619 (2002). |
| Lu et al., “OligoWalk: an online siRNA design tool utilizing hybridization thermodynamics,” Nucleic Acids Research, 36:W104-W108 (2008). |
| Lu et al., “RNA silencing in plants by the expression of siRNA duplexes,” Nucleic Acids Res., 32(21):e171 (2004). |
| Luft, “Making sense out of antisense oligodeoxynucleotide delivery: getting there is half the fun,” J Mol Med, 76:75-76 (1998). |
| Luque et al., “Water Permeability of Isolated Cuticular Membranes: A Structural Analysis,” Archives of Biochemistry and Biophysics, 317(2):417-422 (1995). |
| Maas et al., “Mechanism and optimized conditions for PEG mediated DNA transfection into plant protoplasts,” Plant Cell Reports, 8:148-149 (1989). |
| MacKenzie et al., “Transgenic Nicotiana debneyii expressing viral coat protein are resistant to potato virus S infection,” Journal of General Virology, 71:2167-2170 (1990). |
| Maher III et al., “Inhibition of DNA binding proteins by oligonucleotide-directed triple helix formation,” Science, 245(4919):725-730 (1989). |
| Makkouk el al., “Virus Diseases of Peas, Beans, and Faba Bean in the Mediterranean region,” Adv Virus Res, 84:367-402 (2012). |
| Mandal et al., “Adenine riboswitches and gene activation by disruption of a transcription terminator,” Nature Struct. Mol. Biol., 11(1):29-35 (2004). |
| Mandal et al., “Gene Regulation by Riboswitches,” Nature Reviews Molecular Cell Biology, 5:451-463 (2004). |
| Manoharan, “Oligonucleotide Conjugates as Potential Antisense Drugs with Improved Uptake, Biodistribution, Targeted Delivery, and Mechanism of Action,” Antisense & Nucleic Acid Drug Development, 12:103-128 (2002). |
| Maori et al., “IAPV, a bee-affecting virus associated with Colony Collapse Disorder can be silenced by dsRNA ingestion,” Insect Molecular Biology, 18(1):55-60 (2009). |
| Masoud et al., “Constitutive expression of an inducible β-1,3-glucanase in alfalfa reduces disease severity caused by the oomycete pathogen Phytophthora megasperma f. sp medicaginis, but does not reduce disease severity of chitincontaining fungi,” Transgenic Research, 5:313-323 (1996). |
| Matveeva et al., “Prediction of antisense oligonucleotide efficacy by in vitro methods,” Nature Biotechnology, 16:1374-1375 (1998). |
| Meinke, et al., “Identifying essential genes in Arabidopsis thaliana,” Trends Plant Sci., 13(9):483-491 (2008). |
| Meins et al., “RNA Silencing Systems and Their Relevance to Plant Development,” Annu. Rev. Cell Dev. Biol., 21:297-318 (2005). |
| Melnyk et al., “Intercellular and systemic movement of RNA silencing signals,” The EMBO Journal, 30:3553-3563 (2011). |
| Migge et al., “Greenhouse-grown conditionally lethal tobacco plants obtained by expression of plastidic glutamine synthetase antisense RNA may contribute to biological safety,” Plant Science 153:107-112 (2000). |
| Misawa et al., “Expression of an Erwinia phytoene desaturase gene not only confers multiple resistance to herbicides interfering with carotenoid biosynthesis but also alters xanthophyll metabolism in transgenic plants,” The Plant Journal, 6(4):481-489 (1994). |
| Misawa et al., “Functional expression of the Erwinia uredovora carotenoid biosynthesis gene crtl in transgenic plants showing an increase of β-carotene biosynthesis activity and resistance to the bleaching herbicide norflurazon,” The Plant Journal, 4(5):833-840 (1993). |
| Miura et al., “The Balance between Protein Synthesis and Degradation in Chloroplasts Determines Leaf Variegation in Arabidopsis yellow variegated Mutants,” The Plant Cell, 19:1313-1328 (2007). |
| Molina et al., “Inhibition of protoporphyrinogen oxidase expression in Arahidopsis causes a lesion-mimic phenotype that induces systemic acquired resistance,” The Plant Journal, 17(6):667-678 (1999). |
| Molnar et al., “Plant Virus-Derived Small Interfering RNAs Originate redominantly from Highly Structured Single-Stranded Viral RNAs,” Journal of Virology, 79(12):7812-7818 (2005). |
| Molnar et al., “Small Silencing RNAs in Plants are Mobile and Direct Epigenetic Modification in Recipient Cells,” Science, 328:872-875 (2010). |
| Mora et al., “How Many Species are There on Earth and in the Ocean?,” PLOS Biol., 9(8):e100127, p. 1-8 (2011). |
| Moriyama et al., “Double-stranded RNA in rice: a novel RNA replicon in plants,” Molecular & General Genetics, 248(3):364-369 (1995). |
| Moriyama et al., “Stringently and developmentally regulated levels of a cytoplasmic double-stranded RNA and its high-efficiency transmission via egg and pollen in rice,” Plant Molecular Biology, 31:713-719 (1996). |
| Morrissey el al., “Potent and persistent in vivo anti-HBV activity of chemically modified siRNAs,” Nat Biotechnol. 23(8):1002-1007 (2005). |
| Moser et al., “Sequence-Specific Cleavage of Double Helical DNA by Triple Helix Formation,” Science, 238:645-646 (1987). |
| Mount et al., “Gene and Metabolite Regulatory Network Analysis of Early Developing Fruit Tissues Highlights New Candidate Genes for the Control of Tomato Fruit Composition and Development,” Plant Physiology, 149:1505-1528 (2009). |
| Non-Final Office Action dated Apr. 11, 2013, in U.S. Appl. No. 13/042,856. |
| Non-Final Office Action dated Aug. 19, 2016, in U.S. Appl. No. 13/612,925. |
| Non-Final Office Action dated Aug. 19, 2016, in U.S. Appl. No. 13/612,929. |
| Non-Final Office Action dated Apr. 29, 2016, in U.S. Appl. No. 13/583,302. |
| Non-Final Office Action dated Aug. 10, 2016, in U.S. Appl. No. 13/612,995. |
| Non-Final Office Action dated Aug. 12, 2015, in U.S. Appl. No. 13/612,936. |
| Non-Final Office Action dated Nov. 9, 2016, in U.S. Appl. No. 14/901,003. |
| Non-Final Office Action dated Aug. 13, 2015, in U.S. Appl. No. 13/612,929. |
| Non-Final Office Action dated Aug. 3, 2016, in U.S. Appl. No. 14/015,715. |
| Non-Final Office Action dated Aug. 5, 2016, in U.S. Appl. No. 14/015,785. |
| Non-Final Office Action dated Aug. 8, 2016, in U.S. Appl. No. 13/612,936. |
| Non-Final Office Action dated Dec. 17, 2015, in U.S. Appl. No. 14/532,596. |
| Non-Final Office Action dated Feb. 10, 2016, in U.S. Appl. No. 13/901,326. |
| Non-Final Office Action dated Feb. 23, 2016, in U.S. Appl. No. 14/603,347. |
| Non-Final Office Action dated Feb. 23, 2016, in U.S. Appl. No. 14/608,951. |
| Non-Final Office Action dated Sep. 6, 2016, in U.S. Appl. No. 14/335,135. |
| Non-Final Office Action dated Jul. 23, 2015, in U.S. Appl. No. 14/335,135. |
| Non-Final Office Action dated Jul. 30, 2014, in U.S. Appl. No. 13/042,856. |
| Non-Final Office Action dated Jun. 5, 2015, in U.S. Appl. No. 13/612,948. |
| Non-Final Office Action dated Jun. 8, 2015, in U.S. Appl. No. 13/612,941. |
| Non-Final Office Action dated Mar. 1, 2016, in U.S. Appl. No. 13/612,954. |
| Non-Final Office Action dated Mar. 30, 2015, in U.S. Appl. No. 13/583,302. |
| Non-Final Office Action dated May 15, 2015, in U.S. Appl. No. 14/608,951. |
| Non-Final Office Action dated May 22, 2015, in U.S. Appl. No. 13/612,985. |
| Non-Final Office Action dated Oct. 3, 2016, in U.S. Appl. No. 14/403,491. |
| Non-Final Office Action dated Sep. 1, 2015, in U.S. Appl. No. 13/612,954. |
| Non-Final Office Action dated Sep. 11, 2015, in U.S. Appl. No. 13/612,925. |
| Non-Final Office Action dated Sep. 4, 2015, in U.S. Appl. No. 13/612,995. |
| Nookaraju et al., “Molecular approaches for enhancing sweetness in fruits and vegetables,” Scientia Horticulture, 127:1-15 (2010). |
| Nord-Larsen et al., “Cloning, characterization and expression analysis of tonoplast intrinsic proteins and glutamine synthetase in rycgrass (Lolium perenne L.),” Plant Cell Reports, 28(10):1549-1562 (2009). |
| Notice of Allowance dated Apr. 11, 2016, in U.S. Appl. No. 13/612,985. |
| Notice of Allowance dated Apr. 19, 2016, in U.S. Appl. No. 13/612,941. |
| Notice of Allowance dated Apr. 20, 2016, in U.S. Appl. No. 13/612,948. |
| Notice of Allowance dated Feb. 23, 2015, in U.S. Appl. No. 13/042,856. |
| Notice of Allowance dated Jun. 2, 2015, in U.S. Appl. No. 13/042,856. |
| Notice of Allowance dated Oct. 5, 2015, in U.S. Appl. No. 13/583,302. |
| Nowak et al., “A new and efficient method for inhibition of RNA viruses by DNA interference,” The FEBS Journal, 276:4372-4380 (2009). |
| Office Action dated Apr. 13,2016, in Chinese Patent Application No. 201280053985.3. |
| Office Action dated Aug. 28, 2013, in Chinese Patent Application No. 201180012795.2. |
| Office Action dated Jul. 18, 2016, in Indonesian Patent Application No. W00201203610. |
| Office Action dated Sep. 5, 2016, in Ukrainian Patent Application No. A 2014 03846. |
| Office Action dated Aug. 25, 2016, in Eurasian Patent Application No. 201201264. |
| Office Action dated Feb. 17, 2014, in Mexican Patent Application No. MX/a/2012/010479. |
| Office Action dated Feb. 24, 2014, in Eurasian Patent Application No. 201201264. |
| Office Action dated Jul. 23, 2015, in Ukrainian Patent Application No. 201211548. |
| Office Action dated Jun. 20, 2016, in Chinese Patent Application No. 201280054819.5. |
| Office Action dated Jun. 24, 2016, in Chinese Patent Application No. 201280053984.9. |
| Office Action dated Nov. 3, 2014, in Chinese Patent Application No. 201180012795.2. |
| Office Action dated Jan. 6, 2015, in Japanese Patent Application No. 2012-557165. |
| Office Action dated Nov. 19, 2014, in Eurasian Patent Application No. 201201264/28. |
| Office Action dated Oct. 5, 2015, in Eurasian Patent Application No. 201201264/28. |
| Ongvarrasoponc et al., “A Simple and Cost Effective Method to Generate dsRNA for RNAi Studies in Invertebrates,” Science Asia, 33:35-39 (2007). |
| Orbović et al., “Foliar-Applied Surfactants and Urea Temporarily Reduce Carbon Assimilation of Grapefruit Leaves,” J. Amer. Soc. Hort. Sci., 126(4):486-490 (2001). |
| Ouellet et al., “Members of the Acetohydroxyacid Synthase Muligene Family of Brassica napus Have Divergent Patterns of Expression,” The Plant Journal, Blackwell Scientific Publications, Oxford, GB, 2(3):321-330 (1992). |
| Paddison et al., “Stable suppression of gene expression by RNAi in mammalian cells,” Proc. Natl Acad. Sci. USA, 99(3):1443-1448 (2002). |
| Palauqui et al., “Activation of systemic acquired silencing by localised introduction of DNA,” Current Biology, 9:59-66 (1999). |
| Parera et al., “Dehydration Rate after Solid Matrix Priming Alters Seed Performance of Shrunken-2 Corn,” J. Amer. Soc. Hort. Sci., 119(3):629-635 (1994). |
| Partial Supplementary European Search Report dated Mar. 2, 2015, in European Patent Application No. 12 831 494.5. |
| Patent Examination Report No. 1 dated Feb. 8, 2016, in Australian Patent Application No. 2014262189. |
| Patent Examination Report No. 1 dated Jun. 17, 2016, in Australian Patent Application No. 2012308659. |
| Patent Examination Report No. 1 dated Jun. 17, 2016, in Australian Patent Application No. 2012308660. |
| Patent Examination Report No. 1 dated Nov. 11, 2013, in Australian Patent Application No. 2011224570. |
| Paungfoo-Lonhienne et al., “DNA is Taken up by Root Hairs and Pollen, and Stimulates Root and Pollen Tube Growth,” Plant Physiology, 153:799-805 (2010). |
| Paungfoo-Lonhienne et al., “DNA uptake by Arabidopsis induces changes in the expression of CLE peptides which control root morphology,” Plant Signaling & Behavior, 5(9):1112-1114 (2010). |
| Pei et al., “On the art of identifying effective and specific siRNAs,” Nature Methods, 3(9):670-676 (2006). |
| Peretz et al., “A Universal Expression/Silencing Vector in Plants,” Plant Physiology, 145:1251-1263 (2007). |
| Pornprom et al., “Glutamine synthetase mutation conferring target-site-based resistance to glufosinate in soybean cell selections,” Pest Manag Sci, 2009; 65(2):216-222 (2009). |
| Pratt el al., “Amarantlms rudis and A. tuberculalus, One Species or Two?,” Journal of the Torrey Botanical Society, 128(3):282-296 (2001). |
| Preston et al., “Multiple effects of a naturally occurring proline to threonine substitution within acetolactate synthase in two herbicide-resistant populations of Lactuca serriola,” Pesticide Biochem. Physiol., 84(3):227-235 (2006). |
| Promoter Prediction for SEQ ID No. 1702 from 13/612929/MK/, Promoter 2.0 Prediction Results, pp. 1-4 (2016). |
| Promoter Prediction for SEQ ID No. 4 from 13/612995/MK/, Promoter 2.0 Prediction Results, pp. 1-3 (2016). |
| Promoter Prediction for SEQ ID No. 7 from 13/612936/MK/, Promoter 2.0 Prediction Results, pp. 1-2 (2016). |
| Promoter Prediction for SEQ ID No. 8 from 13/612,925/MK/, Promoter 2.0 Prediction Results, pp. 1-6 (2016). |
| Qiwei,“Advance in DNA interference,” Progress in Veterinary Medicine, 30(1):71-75 (2009). |
| Rajur et al., “Covalent Protein-Oligonucleotide Conjugates for Efficient Delivery of Antisense Molecules,” Bioconjug Chem., 8:935-940 (1997). |
| Reddy et al “Organosilicone Adjuvants Increased the Efficacy of Glyphosate for Control of Weeds in Citrus (Citrus spp.)” HortScience 27(9):1003-1005 (1992). |
| Reddy et al., “Aminomethylphosphonic Acid Accumulation in Plant Species Treated with Glyphosate,” J. Agric. Food Chem., 56(6):2125-2130 (2008). |
| Reither el al., “Specificity of DNA triple helix formation analyted by a FRET assay.” BMC Biochemistry, 3:27 (2002). |
| Restriction Requirement dated Apr. 21, 2015, in U.S. Appl. No. 13/612,954. |
| Restriction Requirement dated Feb. 12, 2015, in U.S. Appl. No. 13/612,985. |
| Restriction Requirement dated Jul. 15, 2016, in U.S. Appl. No. 14/143,748. |
| Restriction Requirement dated Jul. 18, 2016, in U.S. Appl. No. 14/143,836. |
| Restriction Requirement dated Mar. 12, 2015, in U.S. Appl. No. 13/612,948. |
| Restriction Requirement dated Mar. 4, 2015, in U.S. Appl. No. 13/612,941. |
| Restriction Requirement dated May 4, 2015, in U.S. Appl. No. 13/612,929. |
| Restriction Requirement dated May 5, 2015, in U.S. Appl. No. 13/612,936. |
| Restriction Requirement dated May 7, 2015, in U.S. Appl. No. 13/612,925. |
| Restriction Requirement dated May 7, 2015, in U.S. Appl. No. 13/612,995. |
| Restriction Requirement dated Oct. 2, 2012, in U.S. Appl. No. 13/042,856. |
| Restriction Requirement dated Oct. 13, 2016, in U.S. Appl. No. 14/206,707. |
| Restriction Requirement dated Oct. 21, 2014, in U.S. Appl. No. 13/583,302. |
| Restriction Requirement dated Oct. 28, 2015, in U.S. Appl. No. 14/603,347. |
| Restriction Requirement dated Sep. 2, 2015, in U.S. Appl. No. 14/532,596. |
| Rey et al., “Diversity of Dicotyledonous-Infecting Geminiviruses and Their Associated DNA Molecules in Southern Africa, Including the South-West Indian Ocean Islands,” Viruses, 4:1753-1791 (2012). |
| Reynolds et al., “Rational siRNA design for RNA interference,” Nature Biotechnology, 22:326-330 (2004). |
| Riggins et al., “Characterization of de novo transcriptome for waterhemp (Amaranthus tuberculatus) using GS-FLX 454 pyrosequencing and its application for studies of herbicide target-site genes,” Pest Alanag. Sci., 66:1042-1052 (2010). |
| Roberts, “Fast-track applications: The potential for direct delivery of proteins and nucleic acids to plant cells for the discovery of gene function,” Plant Methods, 1(12):1-3 (2005). |
| Robson et al., “Leaf senescence is delayed in maize expressing the Agrobacterium IPT gene under the control of a novel maize senescence-enhanced promoter,” Plant Biotechnology Journal, 2:101-112 (2004). |
| Roitsch et al., “Extracellular invertase: key metabolic enzyme and PR protein,” Journal of Experimental Botany, 54(382):513-524 (2003). |
| Roitsch et al., “Function and regulation of plant invertases: sweet sensations,” Trades in Plant Science, 9(12):606-613 (2004). |
| Rose et al., “Functional polarity is introduced by Dicer processing of short substrate RNAs,” Nucleic Acids Research, 33(13):4140-4156 (2005). |
| Rothnie et al., Pararetroviruses and Retroviruses: A Comparative Review of Viral Structure and Gene Expression Strategies, Advances in Virus Research, 44:1-67 (1994). |
| Ruan et al., “Suppression of Sucrose Synthase Gene Expression Represses Cotton Fiber Cell Initiation, Elongation, and Seed Development,” The Plant Cell, 15:952-964 (2003). |
| Ryabov et al., “Cell-to-Cell, but not Long-Distance, Spread of RNA Silencing That is Induced in Individual Epidermal Cells,” Journal of Virology, 78(6):3149-3154 (2004). |
| Ryan, “Human endogenous retroviruses in health and disease: a symbiotic perspective,” Journal of the Royal Society of Medicine, 97:560-565 (2004). |
| Salanenka et al., “Seedcoat Permeability: Uptake and Post-germination Transport of Applied Model Tracer Compounds,” HortScience, 46(4):622-626 (2011). |
| Santoro et al., “A general purpose RNA-cleaving DNA enzyme,” Proc. Natl. Acad. Sci. USA, 94:4262-4266 (1997). |
| Sathasivan et al., “Nucleotide sequence of a mutant acetolactate synthase gene from an imidazolinone-resistant Arabidopsis thaliana var. Columbia,” Nucleic Acids Research, 18(8):2188-2193 (1990). |
| Schönherr, “Water Permeability of Isolated Cuticular Membranes: The Effect of pH and Cations on Diffusion, Hydrodynamic Permeability and Size of Polar Pores in the Cutin Matrix,” Planta, 128:113-126 (1976). |
| Schwab et al., “RNA silencing amplification in plants: Size matters,” PNAS, 107(34):14945-14946 (2010). |
| Schweizer et al., “Double-stranded RNA interferes with gene function at the single-cell level in cereals,” The Plant Journal, 24(6):895-903 (2000). |
| Schwember et al., “Drying Rates following Priming Affect Temperature Sensitivity of Germination and Longevity of Lettuce Seeds,” HortScience, 40(3):778-781 (2005). |
| Scott et al., Botanical Insecticides for Controlling Agricultural Pests: Piperamides and the Colorado Potato Beetle Leptinotarsa deceinlineata Say (Coleoptera: Chrysomelidae), Archives of Insect Biochemistry and Physiology, 54:212-225 (2003). |
| Second Chinese Office Action dated Jun. 10, 2014, in Chinese Patent Application No. 201180012795.2. |
| Second Office Action dated Feb. 25, 2016, in Chinese Patent Application No. 201280054179.8. |
| Second Office Action dated Mar. 4, 2016, in Chinese Patent Application No. 201280054820.8. |
| Seidman et al., “The potential for gene repair via triple helix formation,” J Clin Invest., 112(4):487-494 (2003). |
| Selvarani et al., “Evaluation of seed priming methods to improve seed vigour of onion (Allium cepa cv. Aggregatum) and carrot (Daucus cetrotet),” Journal of Agricultural Technology, 7(3):857-867 (2011). |
| Senthil-Kumar et al., “A systematic study to determine the extent of gene silencing in Nicotiana benthamiana and other Solanaceae species when heterologous gene sequences are used for virus-induced gene silencing,” New Phytologist, 176:782-791 (2007). |
| Sharma et al., “A simple and efficient Agrobacterium-mediated procedure for transformation of tomato,” J. Biosci., 34(3):423 433 (2009). |
| Shintani et al., “Antisense Expression and Overexpression of Biotin Carboxylase in Tobacco Leaves,” Plant Physiol., 114:881-886 (1997). |
| Showalter, “Structure and Function of Plant Cell Wall Protcins ” The Plant Cell, 5:9-23 (1993). |
| Sijen et al., “On the Role of RNA Amplification in dsRNA-Triggered Gene Silencing,” Cell, 107:465-476 (2001). |
| Silwet L-77 Spray Adjuvant for agricultural applications, product description from Momentive Performance Materials, Inc. (2003). |
| Singh et al., “Absorption and translocation of glyphosate with conventional and organosilicone adjuvants,” Weed Biology and Management, 8:104-111 (2008). |
| Snead et al., “Molecular basis for improved gene silencing by Dicer substrate interfering Rna compared with other siRNA variants,” Nucleic Acids Research, 41(12):6209-6221 (2013). |
| Song et al., “Herbicide,” New Heterocyclic Pesticide, Chemical Industry Press, 354-356 (2011). |
| Steeves et al., “Transgenic soybeans expressing siRNAs specific to a major sperm protein gene suppress Heterodera glycines reproduction,” Funct. Plant Biol., 33:991-999 (2006). |
| Stevens et al., “New Formulation Technology—Silwet® Organosilicone Surfactants Have Physical and Physiological Properties Which Enhance the Performance of Sprays,” Proceedings of the 9th Australian Weeds Conference, pp. 327-331 (1990). |
| Stevens, “Organosilicone Surfactants as Adjuvants for Agrochemicals,” Journal of Pesticide Science, 38:103-122 (1993). |
| Stock et al., “Possible Mechanisms for Surfactant-Induced Foliar Uptake of Agrochemicals,” Pestic. Sci., 38:165-177 (1993). |
| Strat et al., “Specific and nontoxic silencing in mammalian cells with expressed long dsRNAs,” Nucleic Acids Research, 34(13):3803-3810 (2006). |
| Street, “Why is DNA (and not RNA) a stable storage form for genetic information?,” Biochemistry Revisited, pp. 1-4 (2008). |
| Sudarsan et al., “Metabolite-binding RNA domains are present in the genes of eukaryotes,” RNA, 9:644-647 (2003). |
| Sun et al., “A Highly efficient Transformation Protocol for Micro-Tom, a Model Cultivar for Tomato Functional Genomics,” Plant Cell Physiol., 47(3):426-431 (2006). |
| Sun et al., “Antisense oligodeoxynucleotide inhibition as a potent strategy in plant biology: identification of SUSIBA2 as a transcriptional activator in plant sugar signalling,” The Plant Journal, 44:128-138 (2005). |
| Sun et al., “Sweet delivery—sugar translocators as ports of entry for antisense oligodeoxynucleotides in plant cells,” The Plant Journal, 52:1192-1198 (2007). |
| Sutton el al., “Activity of mesotrione on resistant weeds in maize,” Pest Manag. Sci., 58:981-984 (2002). |
| Takasaki et al., “An Effective Method for Selecting siRNA Target Sequences in Mammalian Cells,” Cell Cycle, 3:790-795 (2004). |
| Tang et al., “Efficient delivery of small interfering RNA to plant cells by a nanosecond pulsed laser-induced stress wave for posttranscriptional gene silencing,” Plant Science, 171:375-381 (2006). |
| Tank Mixing Chemicals Applied to Peanut Crops: Are the Chemicals Compatible?, College of Agriculture & Life Sciences, NC State University, AGW-653, pp. 1-11 (2004). |
| Taylor, “Seed Storage, Germination and Quality,” The Physiology of Vegetable Crops, pp. 1-36 (1997). |
| Temple et al., “Can glutamine synthetase activity levels be modulated in transgenic plants by the use of recombinant DNA technology?” Transgenic Plants and Plant Biochemistry, 22(4):915-920 (1994). |
| Temple et al., “Down-regulation of specific members of the glutamine synthetase gene family in Alfalfa by antisense RNA technology,” Plant Molecular Biology, 37:535-547 (1998). |
| Templeton et al., “Improved DNA: liposome complexes for increased systemic delivery and gene expression,” Nature Biotechnology, 15:647-652 (1997). |
| Tenllado et al., “Crude extracts of bacterially expressed dsRNA can be used to protect plants against virus infection,” BMC Biotechnology, 3(3):1-11 (2003). |
| Tenllado et al., “Double-Stranded RNA-Mediated Interference with Plant Virus Infection,” Journal of Virology, 75(24):12288-12297 (2001). |
| Tenllado el al., “RNA interference as a new biotechnological tool for the control of virus diseases in plants,” Virus Research, 102:85-96 (2004). |
| Tepfer, “Risk assessment of virus resistant transgenic plants,” Annual Review of Phytopathology, 40:467-491 (2002). |
| The Seed Biology Place, Website Gerhard Leubner Lab Royal Holloway, University of London, <http://www.seedbiology.de/seedtechnology.asp. |
| Third Party Submission filed on Nov. 29, 2012 in U.S. Appl. No. 13/042,856. |
| Thomas et al., “Size constraints for targeting post-transcriptional gene silencing and for RNA-directed methylation in Nicotiana benthamiana using a potato virus X vector,” The Plant Journal, 25(4):417-425 (2001). |
| Thompson, et al., “Clustal W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice,” Nucl. Acids Res., 22(22):4673-4680 (1994). |
| Timmons et al., “Specific interference by ingested dsRNA,” Nature, 395:854 (1998). |
| Tomari et al., “Perspective: machines for RNAi,” Genes & Dev., 19:517-529 (2005). |
| Tomlinson et al., “Evidence that the hexose-to-sucrose ratio does not control the switch to storage product accumulation in oilseeds: analysis of tobacco seed development and effects of overexpressing apoplastic invertase,” Journal of Experimental Botany, 55(406):2291-2303 (2004). |
| Töpfer et al., “Uptake and Transient Expression of Chimeric Genes in Seed-Derived Embryos,” Plant Cell, 1:133-139 (1989). |
| Toriyama el al., “Transgenic Rice Plants After Direct Gene Transfer Into Protoplasts,” Bio/Technology, 6:1072-1074 (1988). |
| Tran et al., “Control of specific gene expression in mammalian cells by co-expression of long complementary RNAs,” FEBS Lett.;573(1-3):127-134 (2004). |
| Tranel et al., “Resistance of weeds to ALS-inhibiting herbicides: what have we learned?,” Weed Science, 50:700-712 (2002). |
| Tsugawa et al., “Efficient transformation of rice protoplasts mediated by a synthetic polycationic amino polymer,” Theor Appl Genet, 97:1019-1026 (1998). |
| Turina et al., “Tospoviruscs in the Mediterranean Area,” Advances in Virus Research, 84:403-437 (2012). |
| Tuschl, “Expanding small RNA interference,” Nature Biotechnol., 20: 446-448 (2002). |
| Tuschl, “RNA Interference and Small Interfering RNAs,” ChemBiochem. 2(4):239-245 (2001). |
| Ui-Tei et al., “Guidelines for the selection of highly effective siRNA sequences for mammalian and chick RNA interference,” Nucleic Acids Res., 32(3): 936-948 (2004). |
| Unnamalai et al., “Cationic oligopeptide-mediated delivery of dsRNA for post-transcriptional gene silencing in plant cells,” FEBS Letters, 566:307-310 (2004). |
| Unniraman et al., “Alternate Paradigm for Intrinsic Transcription Termination in Eubacteria,” The Journal of Biological Chemistry 276(45)(9):41850-41855 (2001). |
| Unniraman el al., “Conserved Economics of Transcription Termination in Eubacteria,” Nucleic Acids Research, 30(3):675-684 (2002). |
| Urayama et al., “Knock-down of OsDCL2 in Rice Negatively Affects Maintenance of the Endogenous dsRNA Virus, Oryza sativa Endomavirus,” Plant and Cell Physiology, 51(1):58-67 (2010). |
| van de Wetering et al., “Specific inhibition of gene expression using a stably integrated, inducible small-interfering-RNA vector,” EMBO Rep., 4(6):609-615 (2003). |
| Vasil et al., “Herbicide Resistant Fertile Transgenic Wheat Plants Obtained by Microprojectile Bombardment of Regenerable Embryogenic Callus,” Bio/Technology,10:667-674 (1992). |
| Vaucheret, “Post-transcriptional small RNA pathways in plants: mechanisms and regulations,” Genes Dev., 20:759-771 (2006). |
| Vencill et al., “Resistance of Weeds to Herbicides,” Herbicides and Environment, 29:585-594 (2011). |
| Verma et al., “Modified oligonucleotides: synthesis and strategy for users,” Annu. Rev. Biochem., 67:99-134 (1998). |
| Vermeulen et al., “The contributions of dsRNA structure to Dicer specificity and efficiency,” RNA, 11(5):674-682 (2005). |
| Vert et al., “An accurate and interpretable model for siRNA efficacy prediction,” BMC Bioinformatics, 7:520 (2006). |
| Voinnet et al., “Systemic Spread of Sequence-Specific Transgene RNA Degradation in Plants is Initiated by Localized Introduction of Ectopic Promoterless DNA,” Cell, 95:177-187 (1998). |
| Wakelin el al., “A target-site mutation is present in a glyphosate-resistant Lolium rigidum population,” Weed Res. (Oxford), 46(5):432-440 (2006). |
| Walton et al., “Prediction of antisense oligonucleotide binding affinity to a structured RNA target,” Biotechnol Bioeng 65(1):1-9 (1999). |
| Wan et al., “Generation of Large Numbers of Independently Transformed Fertile Barley Plants,” Plant Physiol., 104:37-48 (1994). |
| Wang et al., “Foliar uptake of pesticides-Present status and future challenge,” ScienceDirect, 87:1-8 (2007). |
| Wardell, “Floral Induction of Vegetative Plants Supplied a Purified Fraction of Deoxyribonucleic Acid from Stems of Flowering Plants,” Plant Physiol, 60:885-891 (1977). |
| Wardell, “Floral Activity in Solutions of Deoxyribonucleic Acid Extracted from Tobacco Stems,” Plant Physiol, 57:855-861 (1976). |
| Waterhouse et al., “Virus resistance and gene silencing in plants can be induced by simultaneous expression of sense and antisense RNA,” Proc Natl Acad Sci USA, 95 13959-13964 (1998). |
| Welch et al., “Expression of ribozymes in gene transfer systems to modulate target RNA levels,” Curr Opin Biotechnol. 9(5):486-496 (1998). |
| Widholm et al., “Glyphosate selection of gene amplification in suspension cultures of 3 plant species,” Phyisologia Plantarum, 112:540-545 (2001). |
| Wiesman et al., “Novel cationic vesicle platform derived from vernonia oil for efficient delivery of DNA through plant cuticle membranes,” Journal of Biotechnology, 130:85-94 (2007). |
| Wilson, et al., “Transcription termination at intrinsic terminators: The role of the RNA hairpin,” Proc. Natl. Acad. Sci. USA, 92:8793-8797 (1995). |
| Winkler et al., “Thiamine derivatives bind messenger RNAs directly to regulate bacterial gene expression,” Nature, 419:952-956 (2002). |
| Written Opinion dated Apr. 7, 2016, in Singapore Patent Application No. 201206152-9. |
| Written Opinion dated May 8, 2014, in International Application No. PCT/IL2013/050447. |
| Written Opinion dated Sep. 1, 2014, in Singapore Patent Application No. 201206152-9. |
| Xu et al., Characterization and Functional Analysis of the Calmodulin-Binding Domain of Rac1 GTPase, Plos One, 7(8)1-12:e42975 (2012). |
| Yin et al., “Production of double-stranded RNA for interference with TMV infection utilizing a bacterial prokaryotic expression system,” Appl. Microbiol. Biotechnol., 84(2):323-333 (2009). |
| YouTube video by General Electric Company “Silwet Surfactants,” screen shot taken on Jan. 11, 2012 of video of www.youtube.com/watch?v=WBw7nXMqHk8 (uploaded Jul. 13, 2009). |
| Zagnitko, “Lolium regidum clone LS1 acetyl-CoA carboxylase mRNA, partial cds; nuclear gene for plastid product,” GenBank: AF359516.1, 2 pages (2001). |
| Zagnitko, et al., “An isoleucine/leucine residue in the carboxyltransferase domain of acetyl-CoA carboxylase is critical for interaction with aryloxyphenoxypropionatc and cyclohexanedionc inhibitors,” PNAS, 98(12):6617-6622 (2001). |
| Zhang et al., “A novel rice gene, NRR responds to macronutrient deficiency and regulates root growth,” Mol Plant, 5(1):63-72 (2012). |
| Zhang et al., “Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method,” Nature Protocols, 1(2):1-6 (2006). |
| Zhang et al., “Cationic lipids and polymers mediated vectors for delivery of siRNA,” Journal of Controlled Release, 123:1-10 (2007). |
| Zhang et al., “Chapter 10: New Characteristics of Pesticide Research & Development,” New Progress of the world agriculture chemicals, p. 209 (2010). |
| Zhang et al., “DEG: a database of essential genes,” Nucleic Acids Res., 32:D271-D272 (2004). |
| Zhang et al., “Development and Validation of Endogenous Reference Genes for Expression Profiling of Medaka (Oryzias latipes) Exposed to Endocrine Disrupting Chemicals by Quantitative Real-Time RT-PCR,” Toxicological Sciences, 95(2):356-368 (2007). |
| Zhang et al., “Transgenic rice plants produced by electroporation-mediated plasmid uptake into protoplasts,” The Plant Cell Rep., 7:379-384 (1988). |
| Zhao et al., “Phyllotreta striolata (Coleoptera: Chrysomelidae):Arginine kinase cloning and RNAi-based pest control,” European Journal of Entomology, 105(5):815-822 (2008). |
| Zhu et al., “Ingested RNA interference for managing the populations of the Colorado potato beetle, Leptinotarsa decemlineata,” Pest Manag Sci, 67:175-182 (2010). |
| Anonymous, “Resistant Weeds Spur Research Into New Technologies,” Grains Research & Development Corporation, (2013). |
| Bauer et al., “The major protein import receptor of plastids is essential for chloroplast biogenesis,” Nature, 403:203-207 (2000). |
| Christiaens et al., “The challenge of RNAi-mediated control of hemipterans,” Current Opinion in Insect Science, 6:15-21 (2014). |
| Eudes et al., “Cell-penetrating peptides, Plant Signaling & Behavior,” 3(8):549-5550 (2008). |
| European Search Report dated Sep. 7, 2017, in European Patent Application No. 17152830.0. |
| Extended European Search Report dated Nov. 7, 2017, in European Patent Application No. 15811092.4. |
| Extended European Search Report dated Nov. 8, 2017, in European Patent Application No. 15737282.2. |
| Gan et al., “Bacterially expressed dsRNA protects maize against SCMV infection,” Plant Cell Rep, 29(11):1261-1268 (2010). |
| Gomez-Zurita et al., “Recalibrated Tree of Leaf Beetles (Chrysomelidae) Indicates Independent Diversification of Angiosperms and Their Insect Herbivores,” PLoS One, 4(e360):1-8 (2007). |
| Inaba et al., ““Arabidopsis Tic110 is Essential for the Assembly and Function of the Protein Import Machinery of Plastids,” The Plant Cell, 17:1482-1496 (2005)”. |
| “Jarvis et al, “An Arabidopsis mutant defective in the plastid general protein import apparatus,” Science, 282:100-103 (1998)”. |
| “Kovacheva et al., “Further in vivo studies on the role of the molecular chaperone, Hsp93, in plastid protein import,” The Plant Journal, 50:364-379 (2007)”. |
| “Kovacheva et al., “In vivo studies on the roles of Tic100, Tic40 and Hsp93 during chloroplast protein import,” The Plant Journal, 41:412-428 (2005)”. |
| Office Action dated Dec. 5, 2017, in Japanese Patent Application No. 2016-502033. |
| Partial European Search Report dated Dec. 6, 2017, in European Patent Application No. 17181861.0. |
| Qichuan et al., Seed Science, China Agriculture Press, pp. 101-103, Tables 2-37. |
| Search Report dated Oct. 20, 2017, in Chinese Patent Application No. 201380039346.6. |
| Summons to Attend Oral Proceedings Pursuant to Rule 115(1) EPC, dated Aug. 7, 2017, in European Patent Application No. 12832160.1. |
| Teng et al., “Tic21 is an Essential Translocon Component for Protein Translocation across the Chloroplast Inner Envelope Membrane,” The Plant Cell, 18:2247-2257 (2006). |
| Ulrich et al., “Large scale RNAi screen in Tribolium reveals novel target genes for pest control and the proteasome as prime target,” BMC genomics, 16(1):671 (2015). |
| Xu et al., “Characterization and Functional Analysis of the Calmodulin-Binding Domain of Racl GTPase,” PLoS One, 7(8):e42975 (2012). |
| Zaimin et al., Botany, Northwest A&F University Press, pp. 87-92. |
| Zhong et al., “A pea antisense gene for the chloroplast stromal processing peptidase yields seedling lethals in Arabidopsis: survivors show defective GFP import in vivo,” The Plant Journal, 34:802-812 (2003). |
| Zotti et al., “RNAi technology for insect management and protection of beneficial insects from diseases: lessons, challenges and risk assessments,” Neotropical Entomology, 44(3):197-213 (2015). |
| Baum et al., “Progress Towards RNAi-Mediated Insect Pest Management” Advances in Insect Physiology, 47:249-295 (2014). |
| Burleigh, “Relative Quantitative RT-PCR to Study the Expression of Plant Nutrient Transporters in Athuscular Mycorrhizas” Plant Science, 160:899-904 (2001). |
| Cheng et al., “Transient Expression of Minimum Linear Gene Cassettes in Onion Epidermal Cells Via Direct Transformation,” Appl Biochem Biotechnol, 159:739-749 (2009). |
| Communication Pursuant to Article 94(3) EPC dated Sep. 5, 2018, in European Patent Application No. 17152830.0. |
| Database EMBL XP-002781749(BG442539) dated Mar. 20, 2001. |
| Egli et al., “A Maize Acetyl-Coenzyme A Carboxylase cDNA Sequence,” Plant Physiol., 108: 1299-1300 (1995). |
| European Search Report dated Jun. 29, 2018, in European Patent Application No. 18157745.3. |
| Examination Report dated Mar. 1, 2018, in Australian Patent Application No. 2013264742. |
| Extended European Search Report dated Sep. 28, 2018, in European Patent Application No. 16740770.9. |
| Extended European Search Report dated Apr. 13, 2018, in European Patent Application No. 15812530.0. |
| Extended European Search Report dated Mar. 15, 2018, in European Patent Application No. 17181861.0. |
| Fridlund, “Distribution of Chlorotic Leaf Spot Virus in Apple Budsticks,” Plant Disease Reporter, 57(10): 865-869 (1973). |
| Gasser et al., “Structure, Expression, and Evolution of the 5-Enolpyruvylshikimate-3-phosphate Synthase Genes of Petunia and Tomato,” J. Biol. Chem., 263: 4280-4287 (1988). |
| Gilmer et al., “Latent Viruses of Apple I. Detection with Woody Indictors,” NY St. Agr. Exp. Sta. (Geneva)), 1(10):1-9 (1971). |
| Hoermann et al., “Tic32, as Essential Component in Chloroplast Biogenesis,” The Journal of Biological Chemistry, 279(33):34756-34762 (2004). |
| Hu et al., “High efficiency transport of quantum dots into plant roots with the aid of silwet L-77,” Plant Physiology and Biochemistry, 48:703-709 (2010). |
| Huggett et al., “Real-time RT-PCR Normalisation; Strategies and Considerations,” Genes and Immunity, 6:279-284 (2005). |
| Liu, “Calmodulin and Cell Cycle,” Foreign Medical Sciences Section of Pathophysiology and Clinical Medicine, 18(4):322-324 (1998). |
| Liu, “The Transformation of Nucleic Acid Degradants in Plants,” China Organic Fertilizers, Agriculture Press, ISBN: 7-1091634 (with English translation) (1991). |
| Németh, “The Virus, Mycoplasma and Rickettsia Diseases of Fruit Trees,” Martinus Nijhoff Publishers, The Netherlands and Akadémai Kiadó, Hungary, pp. 197-204 (1986). |
| Non-Final Office Action dated Mar. 21, 2018, in U.S. Appl. No. 13/619,980. |
| Office Action dated Aug. 9, 2018, in Canadian Patent Application No. 2,848,371. |
| Office Action dated Feb. 21, 2018, in Mexican Patent Application No. MX/a/2015/012632 (with English translation). |
| Office Action dated Jul. 30, 2018, in Canadian Patent Application No. 2,848,576. |
| Office Action dated Mar. 8, 2018 (with English translation), in Chilean Patent Application No. 201403192. |
| Office Action dated Sep. 20, 2018, in Chilean Patent Application No. 201601440 (with English translation). |
| Partial Supplementary European Search Report dated Jan. 11, 2018, in European Patent Application No. 15812530.0. |
| Reverdatto et al., “A Multisubunit Acetyl Coenzyme A Carboxylase from Soybean,” Plant Physiol., 119: 961-978 (1999). |
| Schönherr, et al. “Size Selectivity of Aqueous Pores in Astomatous Cuticular Membranes Isolated from Populus canescens (Aiton) Sm. leaves,” Planta 219:405-411 (2004). |
| Stevens, “Formulation of Sprays to Improve the Efficacy of Foliar Fertilisers,” New Zealand Forest Research Institute, pp. 27-34 (1994). |
| Tice, “Selecting the Right Compounds for Screening does Lipinksi's Rule of 5 for Pharmaceuticals Apply to Agrochemicals?,” Pest Manag Sci. 57:3-16 (2001). |
| Asad et al., “Silicon Carbide Whisker-mediated Plant Transformation,” Properties and Applications of Silicon Carbide, pp. 345-358 (2011). |
| Baulcombe, “RNA silencing in plants,” Nature, 431:356-363 (2004). |
| Belhadj et al., “Methyl Jasmonate Induces Defense Responses in Grapevine and Triggers Protection against Erysiphe necator,” J. Agric Food Chem., 54:9119-9125 (2006). |
| Cong et al., “Multiplex Genome Engineering Using CRISPR/Cas Systems,” Science, 339:819-823 (2013). |
| Declaration of Jerzy Zabkiewicz executed Nov. 28, 2017, as filed by Opponent in Australian Patent Application No. 2014262189, pp. 1-73. |
| Declaration of Jerzy Zabkiewicz executed Nov. 28, 2017, as filed by Opponent in Australian Patent Application No. 2014262189, pp. 1-4. |
| Declaration of Neena Mitter executed Nov. 30, 2017, as filed by Opponent in Australian Patent Application No. 2014262189, pp. 1-114. |
| Declaration of Neena Mitter executed Nov. 30, 2017, as filed by Opponent in Australian Patent Application No. 2014262189, pp. 1-25. |
| Delye et al., “PCR-based detection of resistance to acetyl-CoA carboxylase-inhibiting herbicides in black-grass (Alopecurus myosuroides Huds) and ryegrass (Lolium rigidum Gaud),” Pest Management Science, 58:474-478 (2002). |
| Delye et al., “Variation in the gene encoding acetolactate-synthase in Lolium species and proactive detection of mutant, herbicide-resistant alleles,” Weed Research, 49:326-336 (2009). |
| Desveaux et al., “PBF-2 is a Novel Single-Stranded DNA Binding Factor Implicated in PR-10a Gene Activation in Potato,” The Plant Cell, 12:1477-1489 (2000). |
| Dietzgen et al., “Transgenic gene silencing strategies for virus control,” Australasian Plant Pathology, 35:605-618 (2006). |
| Dilpreet et al., “Glyphosate Resistance in a Johnsongrass (Sorghum halepense) Biotype from Arkansas,” Weed Science, 59(3):299-304 (2011). |
| Drobyazko R.V. “Reliable and environmentally friendly insecticide,” Protection and quarantine of plants, 2012 (pp. 52, 53) (in Russian). |
| Duhoux et al., “Reference Genes to Study Herbicide Stress Response in Lolium sp.: Up-Regulation of P3450 Genes in Plants Resistant to Acetolactate-Synthase Inhibitors,” PLOS One, 8(5):e63576 (2013). |
| Extended European Search Report dated Dec. 19, 2018, in European Patent Application No. 16804395.8. |
| Extended European Search Report dated Nov. 16, 2018, in European Patent Application No. 18182238.8. |
| Extended European Search Report dated Nov. 21, 2018, in European Patent Application No. 18175809.5. |
| Gao et al., “DNA-guided genome editing using the Natronobacterium gregoryi Argonaute,” Nature Biotechnology, 34(7):768-773 (2016). |
| Guttieri et al., “DNA Sequence Variation in Domain A of the Acetolactate Synthase Genes of Herbicide-Resistant and -Susceptible Weed Biotypes,” Weed Science, 40:670-679 (1992). |
| Horsch et al., “Inheritance of Functional Foreign Genes in Plants ,” Science, 223:496-498 (1984). |
| Hsu et al., “DNA targeting specificity of RNA-guided Cas9 nucleases,” Nature Biotechnology, 31:827-832 (2013). |
| International Search Report dated Oct. 13, 2016, in International Patent Application No. PCT/US2016/35500. |
| Jiang et al., Chapter III Seeds and Seedlings, Botany, Northwest A&F University Press, pp. 87-92 (2009). |
| Kim et al., “Synthesis and characterization of mannosylated pegylated polyethylenimine as a carrier for siRNA,” International Journal of Pharmaceutics, 427:123-133 (2012). |
| Kirkwood, “Recent developments in our understanding of the plant cuticle as a barrier to the foliar uptake of pesticides,” Pestic Sci, 55:69-77 (1999). |
| Liu, “Confocal laser scanning microscopy—an attractive tool for studying the uptake of xenobiotics into plant foliage,” Journal of Microscopy, 213(Pt 2):87-93 (2004). |
| Lodish et al., Molecular Cell Biology, Fourth Edition, p. 210 (2000). |
| Lucas et al., “Plasmodesmata—bridging the gap between neighboring plant cells,” Trends in Cell Biology, 19:495-503 (2009). |
| Morozov et al., “Evaluation of Preemergence Herbicides for Control of Diclofop-resistant Italian Ryegrass (Lolium multiflorum) in Virginia,” Virginia Polytechnic Institute and State University, pp. 43-71 (2004). |
| N-TER Nanoparticle siRNA, Sigma Aldrich TM website, Web. Nov. 20, 2018. |
| Partial Supplementary European Search Report dated Jan. 11, 2018, in European Patent Application No. 15812530.2. |
| Partial European Search Report dated Jun. 29, 2018, in European Patent Application No. 18157745.3. |
| Pratt et al., “Sorghum Expressed Sequence Tags Identify Signature Genes for Drought, Pathogenesis, and Skotomorphogenesis from a Milestone Set of 16,801 Unique Transcripts,” Plant Physiology, 139:869-884 (2005). |
| Qi et al., “RNA processing enables predictable programming of gene expression,” Nature Biotechnology, 30:1002-1007 (2012). |
| Riar et al., “Glyphosate Resistance in a Johnsongrass (Sorghum halepense) Biotype from Arkansas,” Weed Science, 59:299-304 (2011). |
| Simeoni et al., “Insight into the mechanism of the peptide-based gene delivery system MPG: implications for delivery of siRNA into mammalian cells,” Nucleic Acids Research, 31(11):2717-2724 (2003). |
| Swarts et al., “DNA-guided DNA interference by a prokaryotic Argonaute,” Nature, 507(7491):258-61 (2014). |
| Swarts et al., “Argonaute of the archaeon Pyrococcus furiosus is a DNA-guided nuclease that targets cognate DNA,” Nucleic Acid Res., 43(10):5120-5129 (2015). |
| Townsend et al., “High frequency modification of plant genes using engineered zinc finger nucleases,” Nature, 459:442-445 (2009). |
| TransIT-TKO® Transfection Reagent, Frequently Asked Questions, Web. 2019. |
| Van der Meer et al., “Promoted analysis of the chalcone synthase (chs A) gene of Petunia hybrid: a 67 bp promoter region directs flower-specific expression,” Plant Mol. Biol., 15:95-109 (1990). |
| Vila-Aiub et al., “Glyphosate resistance in perennial Sorghum halepense (Jolmsongrass), endowed by reduced glyphosate translocation and leaf uptake,” Pest Manag Sci, 68:430-436 (2012). |
| Yan et al., Seed Science, China Agriculture Press, pp. 101-103, Tables 2-37 (2001). |
| Yu et al., “Glyphosate, paraquat and ACCase multiple herbicide resistance evolved in a Lolium rigidum biotype,” Planta, 225:499-513 (2007). |
| Yu et al., “Diversity of Acetyl-Coenzyme A Carboxylase Mutations in Resistant Lolium Populations: Evaluation Using Clethodim,” Plant Physiology, 145:547-558 (2007). |
| Zabkiewicz, “Adjuvants and herbicidal efficacy—present status and future prospects,” Weed Research, 40:139-149 (2000). |
| Zhao et al., “Vegetable Standardized Production Technology,” Hangzhou: Zhejiang Science and Technology Press, p. 19 (2008). |
| Zhao et al., “Ps0r1, a potential target for RNA interference-based pest management,” Insect Molecular Biology, 20(1):97-104 (2011). |
| Zidack et al., “Promotion of Bacterial Infection of Leaves by an Organo silicone Surfactant: Implications for Biological Weed Control,” Biological Control, 2:111-117 (1992). |
| Zipperian et al., “Silicon Carbide Abrasive Grinding,” Quality Matters Newsletter, PACE Technologies 1(2):1-3 (2002). |
| Andersen et al., “Delivery of siRNA from lyophilized polymeric surfaces,” Biomaterials, 29:506-512 (2008). |
| Baker, “Chlorophyll Fluorescence: A Probe of Photosynthesis in Vivo,” Annu. Rev. Plant Biol., 59:89-113 (2008). |
| Brugiere et al., “Glutamine Synthetase in the Phloem Plays a Major Role in Controlling Proline Production,” The Plant Cell, 11:195-2011 (1999). |
| Burleigh, “Relative quantitative RT-PCR to study the expression of plant nutrient transporters in arbuscular mycorrhizas,” Plant Science, 160:899-904 (2001). |
| Chang et al., “Dual-target gene silencing by using long, synthetic siRNA duplexes without triggering antiviral responses,” Molecules and Cells, 27(6):689-695 (2009). |
| Communication pursuant to Article 94(3) EPC dated Mar. 16, 2020, in European Patent Application No. 17194281.6. |
| Communication pursuant to Article 94(3) EPC dated Mar. 27, 2020, in European Patent Application No. 15811092.4. |
| Danka et al., “Field Test of Resistance to Acarapis woodi (Acari: Tarsonemidae) and of Colony Production by Four Stocks of Honey Bees (Hymenoptera: Apidae)” Journal of Economic Entomology, 88(3):584-591 (1995). |
| Decision to Grant dated Feb. 24, 2020, in Ukrainian Patent Application No. a 2016 08743 (with English language translation). |
| Declaration of Professor Robert James Henry executed Mar. 1, 2018, as filed by Applicant in Australian Patent Application No. 2014262189, pp. 1-119. |
| Downey et al., “Single and dual parasitic mite infestations on the honey bee, Apis mellifera L.,” Insectes Sociaux, 47(2):171-176 (2000). |
| Drobyazko R.V., “Reliable and environmentally friendly insecticide,” Protection and quarantine of plants, 2012 (pp. 52, 53) (with English translation). |
| Extended European Search Report dated Mar. 25, 2020, in European Patent Application No. 19192942.1. |
| Gilmer et al., “Latent Viruses of Apple I. Detection with Woody Indicators,” Plant Pathology, 1(10):1-9 (1971). |
| Hormann et al., “Tic32, as Essential Component in Chloroplast Biogenesis,” The Journal of Biological Chemistry, 279(33):34756-34762 (2004). |
| Hwa et al., “Fixation of hybrid vigor in rice: opportunities and challenges,” Euphytica, 160:287-293 (2008). |
| Jasieniuk et al., “Glyphosate-Resistant Italian Ryegrass (Lolium multiflorum) in California: Distribution, Response to Glyphosate, and Molecular Evidence for an Altered Target Enzyme,” Weed Science, 56(4):496-502 (2008). |
| Khanbekova et al., The defeat of the honey bee Apis melifera caucasica Gorb. By viruses and parasites, and condition of bee colonies in different ecogeographical conditions of Greater Caucasus, Agricultural Biology. 2013 (p. 43) (in Russian). |
| Li et at., “Long dsRNA but not siRNA initiates RNAi in western corn rootworm larvae and adults,” Journal of Applied Entomology, 139(6):432-445 (2015). |
| Nemeth, “Virus, mycoplasma and rickettsia diseases of fruit trees,” Martinus Nijhoff Publishers, 197-204 (1986). |
| N-TER Nanoparticle siRNA, Sigma Aldrich TM website, Web. Nov. 20, 2018 <https://www.sigmaaldrich.com/life-science/custom-oligos/sirna-oligos/n-ter-nanoparticle.html>. |
| Office Action dated Feb. 20, 2020, in Canadian Patent Application No. 2,905,104. |
| Office Action dated Feb. 25, 2020, in Japanese Patent Application No. 2017-538699 (with English language translation). |
| Ossowski et al., “Gene silencing in plants using artificial microRNAs and other small RNAs,” The Plant Journal, 53:674-690 (2008). |
| Partial European Search Report dated Dec. 6, 2019, in European Patent Application No. 19185431.4. |
| Prado et al., “Design and optimization of degenerated universal primers for the doing of the plant acetolactate synthase conserved domains,” Weed Science, 52:487-491 (2004). |
| Regalado, “The Next Great GMO Debate,” MIT Technology Review, pp. 1-19 (2015) <https://www.technologyreview.com/s/540136/the-next-great-gmo-debate/>. |
| Sammataro et al., “Some Volatile Plant Oils as Potential Control Agents for Varroa Mites (Acari: Varroidae) in Honey Bee Colonies (Hymenoptera: Apidae),” American Bee Journal, 138(9):681-685 (1998). |
| Subramoni et al., “Lipases as Pathogenicity Factors of Plant Pathogens,” Handbook of Hydrocarbon and Lipid Microbiology, 3269-3277 (2010). |
| Sun, “Characterization of Organosilicone Surfactants and Their Effects on Sulfonylurea Herbicide Activity,” Thesis Submitted to the Faculty of the Virginia Polytechnic Institute and State University dated Apr. 5, 1996. |
| Tenllado et al., “Crude extracts of bacterially expressed dsRNA can be used to protect plants against virus infections,” BMC Biotechnology, 3:1-11 (2003). |
| Tice, “Selecting the right compounds for screening: does Lipinski's Rule of 5 for pharmaceuticals apply to agrochemicals?” Pest Management Science, 57(1):3-16 (2001). |
| TransIT-TKO® Transfection Reagent, Frequently Asked Questions, Web. 2019 <https://www.mirusbio.com/tech-resources/fags/transit-tko-faqs>. |
| Walton, “Deconstructing the Cell Wall,” Plant Physiol., 104:1113-1118 (1994). |
| Wang et al., “Principle and technology of genetic engineering in plants,” in Plant genetic engineering principles and techniques, Beijing: Science Press, pp. 313-315 (1998). |
| Watson et al., “RNA silencing platforms in plants,” FEBS Letters, 579:5982-5987 (2005). |
| Wild Carrot, Noxious Weed Control Board (NWCB) of Washington State (2010) <www.nwcb.wa.gov/detail.asp?weed=46>. |
| Yibrah et al., “Antisense RNA inhibition of uidA gene expression in transgenic plants: Evidence for interaction between first and second transformation events,” Hereditas, 118:273-280 (1993). |
| Zhao et al., “Vegetable Statdardized Production Technology,” Hangzhou: Zhejiang Science and Technology Press, p. 19 (2008). |
| Number | Date | Country | |
|---|---|---|---|
| 20160330964 A1 | Nov 2016 | US |
| Number | Date | Country | |
|---|---|---|---|
| 61534076 | Sep 2011 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | 13612948 | Sep 2012 | US |
| Child | 15227651 | US |
