INSECTCIDAL PROTEIN COMBINATIONS FOR CONTROLLING FALL ARMYWORM AND EUROPEAN CORN BORER, AND METHODS FOR INSECT RESISTANCE MANAGEMENT

Abstract
The subject invention relates in part to stacking certain Cry genes along with Cry 1Fa to result in products that are more durable and less prone towards insects developing resistance towards the activity of any of the toxins (such as Cry 1Fa) by itself. Embodiments of the Cry 1F stacking partners include Cry2Aa, Cry 1I, and Cry 1E. These stacks can be used to control FAW and/or ECB as described herein.
Description
BACKGROUND

Humans grow corn for food and energy applications. Insects eat and damage corn plants and thereby undermine these human efforts. Agricultural insect pests causing major destruction of corn are the fall armyworm (FAW, Spodoptera frugiperda) and European corn borer (ECB, Ostrinia nubilalis).


Current in-plant transgenic control of these pests is achieved through plant expression of a crystal (Cry) delta endotoxin gene coding for the Cry1Fa protein from Bacillus thuringiensis. Cry1Fa is the protein toxin currently in the Herculex™ brand of Dow AgroSciences transgenic corn seeds (Herculex, Herculex-Extra, and Herculex-RW) that are resistant to FAW and ECB insect pests. This protein works by binding to specific receptor(s) located in the midgut of insects, and forms pores within the gut cells. The formation of these pores prevents insects from regulating osmotic balance which results in their death.


However, some are concerned that insects might be able to develop resistance to the action of Cry1Fa through genetic alterations of the receptors within their gut that bind Cry 1Fa. Insects that produce receptors with a reduced ability to bind Cry1Fa can be resistant to the activity of Cry1Fa, and thus survive on plants that express this protein.


With a single Cry toxin continuously present in the plant during growth conditions, there is concern that insects could develop resistance to the activity of this protein through genetic alterations of the receptor that binds Cry1Fa toxin in the insect gut. Reductions in toxin binding due to these alterations in the receptor would lead to reduced toxicity of the Cry1Fa possibly leading to eventual decreased effectiveness of the protein when expressed in a crop.


BRIEF SUMMARY

The subject invention relates in part to stacking certain Cry genes along with Cry1Fa to result in products that are more durable and less prone towards insects developing resistance towards the activity of any of the toxins (such as Cry1Fa) by itself. Embodiments of the Cry1F stacking partners include Cry2Aa and/or Cry1I—for targeting fall armyworm (FAW; Spodoptera frugiperda), and Cry1E—for targeting European cornborer (ECB; Ostrinia nubilalis).







DETAILED DESCRIPTION

The subject invention includes the use of at least one of the stacking partner toxins with Cry1Fa toxins as a pair. Some preferred pairs for targeting FAW (fall armyworm; Spodoptera frugiperda) are a Cry1Fa protein plus a Cry1Ea protein. Some preferred pairs for targeting ECB (European cornborer; Ostrinia nubilalis) comprise a Cry1Fa protein plus a Cry1I and/or a Cry2Aa protein.


The subject invention also relates in part to triple stacks or “pyramids” of three (or more) toxins, with Cry1Fa and a stacking partner toxin being the base pair. One preferred pyramid provides two modes of action against two pests—the FAW and the ECB. This type of “2 MOA” pyramid includes: Cry1Fa plus Cry2Aa plus Cry lAb (a preferred embodiment for controlling ECB), and Cry1Fa plus Cry1Ea (a preferred embodiment for controlling FAW). By “separate modes of action” it is meant that the proteins do not cause cross-resistance with each other.


In some preferred pyramid embodiments, the selected toxins provide three separate modes of action (active ingredients that do not result in cross-resistance) against ECB. Preferred pyramid combinations are Cry1Fa plus a second IRM toxin plus a third IRM toxin. Examples of such are as follows.


ECB pyramids of the subject invention include a Cry1Fa toxin plus a Cry2Aa toxin as the second IRM toxin and a third IRM toxin selected from the group consisting of Cry1Be, Cry1Ab, DIG-3, and Cry1I toxins.


ECB pyramids of the subject invention include a Cry1Fa toxin plus a Cry1I toxin as the second IRM toxin and a third IRM toxin selected from the group consisting of Cry1Ab, Cry1Be, DIG-3, and Cry2Aa toxins.


These various toxins (and others) are listed in the attached Appendix A. Those GENBANK numbers can also be used to obtain the sequences for any of the genes and proteins disclosed or mentioned herein.


Patents can also be used to obtain relevant sequences. For example, U.S. Pat. No. 5,188,960 and U.S. Pat. No. 5,827,514 describe Cry1Fa core toxin containing proteins suitable for use in carrying out the present invention. U.S. Pat. No. 6,218,188 describes plant-optimized DNA sequences encoding Cry1Fa core toxin-containing proteins that are suitable for use in the present invention. US 2010-00269223 relates to DIG-3 proteins.


The subject invention also relates generally to the use of three insecticidal proteins (Cry proteins in some preferred embodiments) that do not compete with each other/do not result in cross-resistance yet are active against a single target pest.


Plants (and acreage planted with such plants) that produce any of the subject pairs, or three (or more), toxins are included within the scope of the subject invention. Additional toxins/genes can also be added, but these particular triple stacks would, according to the subject invention, advantageously and surprisingly provide three modes of action (non-cross-resistant activity) against FAW and/or ECB. This can help to reduce or eliminate the requirement for refuge acreage (e.g., less than 40%, less than 20%, less than 10%, less than 5%, or even 0% refuge). A field thus planted of over 10 acres is thus included within the subject invention.


The subject polynucleotide(s) are preferably in a genetic construct under control of (operably linked to/comprising) a non-Bacillus-thuringiensis promoter. The subject polynucleotides can comprise plant codon usage for enhanced expression in a plant.


To counter act the ability of insects to develop resistance to Cry1Fa, we identified Cry toxins that non-competitively (with Cry1Fa) bind to FAW and/or ECB gut cell preparations. Cry1Fa does not to displace binding of Cry proteins identified herein in the insect gut of FAW and ECB larvae—Cry proteins such as Cry2Aa. The ability of these Cry toxins to be toxic to FAW and/or ECB larvae, yet not fully interact with the same sites as Cry1Fa, shows that their toxicity will not be affected by insects having developed genetic alterations of their Cry1Fa receptor as a mechanism to become resistant to the toxicity of Cry1Fa.


Thus insects having developed resistance to Cry1Fa through a reduction in the ability of its gut receptors to bind Cry1Fa would still be susceptible to the toxicity of Cry2Aa proteins, for example, which bind alternative sites. We have obtained biochemical data that supports this. Having combinations of these proteins expressed in transgenic plants thus provides a useful and valuable mechanism to reduce the probability for the development of insect resistance in the field and thus lead towards a reduction in the requirement for refugia. Other Cry proteins have been studied for their activity against other major insect pests, both sensitive, and those resistant to Cry1Fa (rFAW and rECB). As shown in Table 1, Cry1I and Cry2Aa are active against both resistant and susceptible ECB larvae. Cry1Ea is active against both resistant and susceptible FAW. Binding data for these Cry toxins against these insect pests can be obtained. In any case, the data herein described below shows the toxins interacting at separate target site(s) within the insect gut compared to Cry1Fa and thus would make excellent stacking partners.


Stacking Cry1Fa expressing crops with one or more additional Cry genes, such as those described herein result in an effective management strategy to prevent the ability of insects to develop tolerance to the activity of transgenic plants expressing these protein toxins. Since we show that these Cry proteins interact at different and/or overlapping sites compared to Cry1Fa, if resistance were to occur through alterations in the affinity of the insect gut receptors that bind to the Cry toxins, the alteration would have to occur in at least two different receptors simultaneously to allow the insects to survive on plants expressing the multiple proteins. The probability of this occurring is extremely remote, thus increasing the durability of the transgenic product to ward of insects being able to develop tolerance to the proteins.


We radio-iodinated trypsin truncated forms of the Cry protein toxins and used radioreceptor binding assay techniques to measure their binding interaction with putative receptor proteins located within the insect gut membranes. The gut membranes were prepared as brush border membrane vesicles (BBMV) by the method of Wolfersberger. Iodination of the toxins were conducted using either iodo beads or iodogen treated tubes from Pierce Chemicals. Specific activity of the radiolabeled toxin was approximately 1-4 μCi/μg protein. Binding studies were carried out essentially by the procedures of Liang.


The data presented herein shows the toxins interacting at separate target site within the insect gut compared to Cry1Fa and thus would make excellent stacking partners.


The subject invention can be used with a variety of plants. Examples include corn (maize), soybeans, and cotton.


Genes and toxins useful according to the subject invention include not only the full length sequences disclosed but also fragments of these sequences, variants, mutants, and fusion proteins which retain the characteristic pesticidal activity of the toxins specifically exemplified herein. As used herein, the terms “variants” or “variations” of genes refer to nucleotide sequences which encode the same toxins or which encode equivalent toxins having pesticidal activity. As used herein, the term “equivalent toxins” refers to toxins having the same or essentially the same biological activity against the target pests as the claimed toxins.


As used therein, the boundaries represent approximately 95% (e.g. Cry1Fa's), 78% (e.g. Cry1F's), and 45% (Cry1's) sequence identity, per “Revision of the Nomenclature for the Bacillus thuringiensis Pesticidal Crystal Proteins,” N. Crickmore, D. R. Zeigler, J. Feitelson, E. Schnepf, J. Van Rie, D. Lereclus, J. Baum, and D. H. Dean. Microbiology and Molecular Biology Reviews (1998) Vol 62: 807-813. These cut offs can also be applied to the core proteins only (for Cry1F and Cry1Fa core proteins, for example). See related Crickmore et al. website at lifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/.


Fragments and equivalents which retain the pesticidal activity of the exemplified toxins would be within the scope of the subject invention. Also, because of the redundancy of the genetic code, a variety of different DNA sequences can encode the amino acid sequences disclosed herein. It is well within the skill of a person trained in the art to create these alternative DNA sequences encoding the same, or essentially the same, toxins. These variant DNA sequences are within the scope of the subject invention. As used herein, reference to “essentially the same” sequence refers to sequences which have amino acid substitutions, deletions, additions, or insertions which do not materially affect pesticidal activity. Fragments of genes encoding proteins that retain pesticidal activity are also included in this definition.


A further method for identifying the genes encoding the toxins and gene portions useful according to the subject invention is through the use of oligonucleotide probes. These probes are detectable nucleotide sequences. These sequences may be detectable by virtue of an appropriate label or may be made inherently fluorescent as described in International Application No. WO93/16094. As is well known in the art, if the probe molecule and nucleic acid sample hybridize by forming a strong bond between the two molecules, it can be reasonably assumed that the probe and sample have substantial homology. Preferably, hybridization is conducted under stringent conditions by techniques well-known in the art, as described, for example, in Keller, G. H., M. M. Manak (1987) DNA Probes, Stockton Press, New York, N.Y., pp. 169-170. Some examples of salt concentrations and temperature combinations are as follows (in order of increasing stringency): 2×SSPE or SSC at room temperature; 1×SSPE or SSC at 42° C.; 0.1×SSPE or SSC at 42° C.; 0.1×SSPE or SSC at 65° C. Detection of the probe provides a means for determining in a known manner whether hybridization has occurred. Such a probe analysis provides a rapid method for identifying toxin-encoding genes of the subject invention. The nucleotide segments which are used as probes according to the invention can be synthesized using a DNA synthesizer and standard procedures. These nucleotide sequences can also be used as PCR primers to amplify genes of the subject invention.


Certain proteins of the subject invention have been specifically exemplified herein. Since these proteins are merely exemplary of the proteins of the subject invention, it should be readily apparent that the subject invention comprises variant or equivalent proteins (and nucleotide sequences coding for equivalent proteins) having the same or similar pesticidal activity of the exemplified protein. Equivalent proteins will have amino acid homology with an exemplified protein. This amino acid homology will typically be greater than 75%, preferably be greater than 90%, and most preferably be greater than 95%. The amino acid homology will be highest in critical regions of the protein which account for biological activity or are involved in the determination of three-dimensional configuration which ultimately is responsible for the biological activity. In this regard, certain amino acid substitutions are acceptable and can be expected if these substitutions are in regions which are not critical to activity or are conservative amino acid substitutions which do not affect the three-dimensional configuration of the molecule. For example, amino acids may be placed in the following classes: non-polar, uncharged polar, basic, and acidic. Conservative substitutions whereby an amino acid of one class is replaced with another amino acid of the same type fall within the scope of the subject invention so long as the substitution does not materially alter the biological activity of the compound. Following is a listing of examples of amino acids belonging to each class. In some instances, non-conservative substitutions can also be made. The critical factor is that these substitutions must not significantly detract from the biological activity of the protein.
















Class of Amino Acid
Examples of Amino Acids









Nonpolar
Ala, Val, Leu, Ile, Pro, Met, Phe, Trp



Uncharged Polar
Gly, Ser, Thr, Cys, Tyr, Asn, Gln



Acidic
Asp, Glu



Basic
Lys, Arg, His










Insect Resistance Management (IRM) Strategies. Roush et al., for example, outlines two-toxin strategies, also called “pyramiding” or “stacking,” for management of insecticidal transgenic crops. (The Royal Society. Phil. Trans. R. Soc. Lond. B. (1998) 353, 1777-1786).


On their website, the United States Environmental Protection Agency (epa.gov/oppbppd1/biopesticides/pips/bt_corn_refuge2006.htm) publishes the following requirements for providing non-transgenic (i.e., non-B.t.) refuges (a section of non-Bt crops/corn) for use with transgenic crops producing a single Bt protein active against target pests.

    • “The specific structured requirements for corn borer-protected Bt (Cry1Ab or Cry1F) corn products are as follows:
    • Structured refuges: 20% non-Lepidopteran Bt corn refuge in Corn Belt;
      • 50% non-Lepidopteran Bt refuge in Cotton Belt
    • Blocks
      • Internal (i.e., within the Bt field)
      • External (i.e., separate fields within ½ mile (¼ mile if possible) of the Bt field to maximize random mating)
    • In-field Strips
      • Strips must be at least 4 rows wide (preferably 6 rows) to reduce the effects of larval movement”


In addition, the National Corn Growers Association, on their website (ncga.com/insect-resistance-management-fact-sheet-bt-corn) also provides similar guidance regarding the refuge requirements. For example:

    • “Requirements of the Corn Borer IRM:
      • Plant at least 20% of your corn acres to refuge hybrids
      • In cotton producing regions, refuge must be 50%
      • Must be planted within ½ mile of the refuge hybrids
      • Refuge can be planted as strips within the Bt field; the refuge strips must be at least 4 rows wide
      • Refuge may be treated with conventional pesticides only if economic thresholds are reached for target insect
      • Bt-based sprayable insecticides cannot be used on the refuge corn
      • Appropriate refuge must be planted on every farm with Bt corn”


As stated by Roush et al. (on pages 1780 and 1784 right column, for example), stacking or pyramiding of two different proteins each effective against the target pests and with little or no cross-resistance can allow for use of a smaller refuge. Roush suggests that for a successful stack, a refuge size of less than 10% refuge, can provide comparable resistance management to about 50% refuge for a single (non-pyramided) trait. For currently available pyramided Bt corn products, the U.S. Environmental Protection Agency requires significantly less (generally 5%) structured refuge of non-Bt corn be planted than for single trait products (generally 20%).


There are various ways of providing the IRM effects of a refuge, including various geometric planting patterns in the fields (as mentioned above) and in-bag seed mixtures, as discussed further by Roush et al. (supra), and U.S. Pat. No. 6,551,962.


The above percentages, or similar refuge ratios, can be used for the subject double or triple stacks or pyramids. For triple stacks with three sites of action against a single target pest, a goal would be zero refuge (or less than 5% refuge, for example). This is particularly true for commercial acreage—of over 10 acres for example.


All patents, patent applications, provisional applications, and publications referred to or cited herein are incorporated by reference in their entirety to the extent they are not inconsistent with the explicit teachings of this specification.


Unless specifically indicated or implied, the terms “a”, “an”, and “the” signify “at least one” as used herein.


Following are examples that illustrate procedures for practicing the invention. These examples should not be construed as limiting. All percentages are by weight and all solvent mixture proportions are by volume unless otherwise noted. All temperatures are in degrees Celsius.


REFERENCES



  • Wolfersberger, M. G., (1993), Preparation and Partial Characterization of Amino Acid Transporting Brush Border Membrane Vesicles from the Larval Midgut of the Gypsy Moth (Lymantria Dispar). Arch. Insect Biochem. Physiol. 24: 139-147.

  • Liang, Y., Patel, S. S., and Dean, D. H., (1995), Irreversible Binding Kinetics of Bacillus thuringiensis Cry1A Delta-Endotoxins to Gypsy Moth Brush Border Membrane Vesicles is Directly Correlated to Toxicity. J. Biol. Chem., 270, 24719-24724.



Example 1
Bioassay

The subject Cry proteins were studied for their activity against other major insect pests, both sensitive, and those resistant to Cry1Fa (rFAW and rECB). As shown in Table 1, Cry1I and Cry2Aa are active against both resistant and susceptible ECB larvae. Cry1Ea is active against both resistant and susceptible FAW. (For a general discussion of this pest, see e.g. Tabashnik, PNAS (2008), vol. 105 no. 49, 19029-19030.)













TABLE 1









PROTEIN
FAW
rFAW







Cry1Ea
144
1845



Cry2Aa
1397.9
1936.8



Cry1Fa
22
inactive







PROTEIN
ECB
rECB







Cry1I
79.1
3618



Cry2Aa
122.9
188.4



Cry1Fa
16
inactive










APPENDIX A

List of delta-endotoxins—from Crickmore et al. website (cited in application)


Accession Number is to NCBI entry (if available)

















Name
Acc No.
Authors
Year
Source Strain
Comment







Cry1Aa1
AAA22353
Schnepf et al
1985
Bt kurstaki HD1



Cry1Aa2
AAA22552
Shibano et al
1985
Bt sotto


Cry1Aa3
BAA00257
Shimizu et al
1988
Bt aizawai IPL7


Cry1Aa4
CAA31886
Masson et al
1989
Bt entomocidus


Cry1Aa5
BAA04468
Udayasuriyan et al
1994
Bt Fu-2-7


Cry1Aa6
AAA86265
Masson et al
1994
Bt kurstaki NRD-






12


Cry1Aa7
AAD46139
Osman et al
1999
Bt C12


Cry1Aa8
I26149
Liu
1996

DNA sequence only


Cry1Aa9
BAA77213
Nagamatsu et al
1999
Bt dendrolimus






T84A1


Cry1Aa10
AAD55382
Hou and Chen
1999
Bt kurstaki HD-1-






02


Cry1Aa11
CAA70856
Tounsi et al
1999
Bt kurstaki


Cry1Aa12
AAP80146
Yao et al
2001
Bt Ly30


Cry1Aa13
AAM44305
Zhong et al
2002
Bt sotto


Cry1Aa14
AAP40639
Ren et al
2002
unpublished


Cry1Aa15
AAY66993
Sauka et al
2005
Bt INTA Mol-12


Cry1Ab1
AAA22330
Wabiko et al
1986
Bt berliner 1715


Cry1Ab2
AAA22613
Thorne et al
1986
Bt kurstaki


Cry1Ab3
AAA22561
Geiser et al
1986
Bt kurstaki HD1


Cry1Ab4
BAA00071
Kondo et al
1987
Bt kurstaki HD1


Cry1Ab5
CAA28405
Hofte et al
1986
Bt berliner 1715


Cry1Ab6
AAA22420
Hefford et al
1987
Bt kurstaki NRD-






12


Cry1Ab7
CAA31620
Haider & Ellar
1988
Bt aizawai IC1


Cry1Ab8
AAA22551
Oeda et al
1987
Bt aizawai IPL7


Cry1Ab9
CAA38701
Chak & Jen
1993
Bt aizawai HD133


Cry1Ab10
A29125
Fischhoff et al
1987
Bt kurstaki HD1


Cry1Ab11
I12419
Ely & Tippett
1995
Bt A20
DNA sequence only


Cry1Ab12
AAC64003
Silva-Werneck et al
1998
Bt kurstaki S93


Cry1Ab13
AAN76494
Tan et al
2002
Bt c005


Cry1Ab14
AAG16877
Meza-Basso &
2000
Native Chilean Bt




Theoduloz


Cry1Ab15
AAO13302
Li et al
2001
Bt B-Hm-16


Cry1Ab16
AAK55546
Yu et al
2002
Bt AC-11


Cry1Ab17
AAT46415
Huang et al
2004
Bt WB9


Cry1Ab18
AAQ88259
Stobdan et al
2004
Bt


Cry1Ab19
AAW31761
Zhong et al
2005
Bt X-2


Cry1Ab20
ABB72460
Liu et al
2006
BtC008


Cry1Ab21
ABS18384
Swiecicka et al
2007
Bt IS5056


Cry1Ab22
ABW87320
Wu and Feng
2008
BtS2491Ab


Cry1Ab-
AAK14336
Nagarathinam et al
2001
Bt kunthala RX24
uncertain sequence


like


Cry1Ab-
AAK14337
Nagarathinam et al
2001
Bt kunthala RX28
uncertain sequence


like


Cry1Ab-
AAK14338
Nagarathinam et al
2001
Bt kunthala RX27
uncertain sequence


like


Cry1Ab-
ABG88858
Lin et al
2006
Bt ly4a3
insufficient sequence


like


Cry1Ac1
AAA22331
Adang et al
1985
Bt kurstaki HD73


Cry1Ac2
AAA22338
Von Tersch et al
1991
Bt kenyae


Cry1Ac3
CAA38098
Dardenne et al
1990
Bt BTS89A


Cry1Ac4
AAA73077
Feitelson
1991
Bt kurstaki






PS85A1


Cry1Ac5
AAA22339
Feitelson
1992
Bt kurstaki






PS81GG


Cry1Ac6
AAA86266
Masson et al
1994
Bt kurstaki NRD-






12


Cry1Ac7
AAB46989
Herrera et al
1994
Bt kurstaki HD73


Cry1Ac8
AAC44841
Omolo et al
1997
Bt kurstaki HD73


Cry1Ac9
AAB49768
Gleave et al
1992
Bt DSIR732


Cry1Ac10
CAA05505
Sun
1997
Bt kurstaki YBT-






1520


Cry1Ac11
CAA10270
Makhdoom &
1998




Riazuddin


Cry1Ac12
I12418
Ely & Tippett
1995
Bt A20
DNA sequence only


Cry1Ac13
AAD38701
Qiao et al
1999
Bt kurstaki HD1


Cry1Ac14
AAQ06607
Yao et al
2002
Bt Ly30


Cry1Ac15
AAN07788
Tzeng et al
2001
Bt from Taiwan


Cry1Ac16
AAU87037
Zhao et al
2005
Bt H3


Cry1Ac17
AAX18704
Hire et al
2005
Bt kenyae HD549


Cry1Ac18
AAY88347
Kaur & Allam
2005
Bt SK-729


Cry1Ac19
ABD37053
Gao et al
2005
Bt C-33


Cry1Ac20
ABB89046
Tan et al
2005


Cry1Ac21
AAY66992
Sauka et al
2005
INTA Mol-12


Cry1Ac22
ABZ01836
Zhang & Fang
2008
Bt W015-1


Cry1Ac23
CAQ30431
Kashyap et al
2008
Bt


Cry1Ac24
ABL01535
Arango et al
2008
Bt 146-158-01


Cry1Ac25
FJ513324
Guan Peng et al
2008
Bt Tm37-6
No NCBI link July 09


Cry1Ac26
FJ617446
Guan Peng et al
2009
Bt Tm41-4
No NCBI link July 09


Cry1Ac27
FJ617447
Guan Peng et al
2009
Bt Tm44-1B
No NCBI link July 09


Cry1Ac28
ACM90319
Li et al
2009
Bt Q-12


Cry1Ad1
AAA22340
Feitelson
1993
Bt aizawai PS81I


Cry1Ad2
CAA01880
Anonymous
1995
Bt PS81RR1


Cry1Ae1
AAA22410
Lee & Aronson
1991
Bt alesti


Cry1Af1
AAB82749
Kang et al
1997
Bt NT0423


Cry1Ag1
AAD46137
Mustafa
1999


Cry1Ah1
AAQ14326
Tan et al
2000


Cry1Ah2
ABB76664
Qi et al
2005
Bt alesti


Cry1Ai1
AAO39719
Wang et al
2002


Cry1A-
AAK14339
Nagarathinam et al
2001
Bt kunthala nags3
uncertain sequence


like


Cry1Ba1
CAA29898
Brizzard & Whiteley
1988
Bt thuringiensis






HD2


Cry1Ba2
CAA65003
Soetaert
1996
Bt entomocidus






HD110


Cry1Ba3
AAK63251
Zhang et al
2001


Cry1Ba4
AAK51084
Nathan et al
2001
Bt entomocidus






HD9


Cry1Ba5
ABO20894
Song et al
2007
Bt sfw-12


Cry1Ba6
ABL60921
Martins et al
2006
Bt S601


Cry1Bb1
AAA22344
Donovan et al
1994
Bt EG5847


Cry1Bc1
CAA86568
Bishop et al
1994
Bt morrisoni


Cry1Bd1
AAD10292
Kuo et al
2000
Bt wuhanensis






HD525


Cry1Bd2
AAM93496
Isakova et al
2002
Bt 834


Cry1Be1
AAC32850
Payne et al
1998
Bt PS158C2


Cry1Be2
AAQ52387
Baum et al
2003


Cry1Be3
FJ716102
Xiaodong Sun et al
2009
Bt
No NCBI link July 09


Cry1Bf1
CAC50778
Arnaut et al
2001


Cry1Bf2
AAQ52380
Baum et al
2003


Cry1Bg1
AAO39720
Wang et al
2002


Cry1Ca1
CAA30396
Honee et al
1988
Bt entomocidus






60.5


Cry1Ca2
CAA31951
Sanchis et al
1989
Bt aizawai 7.29


Cry1Ca3
AAA22343
Feitelson
1993
Bt aizawai PS81I


Cry1Ca4
CAA01886
Van Mellaert et al
1990
Bt entomocidus






HD110


Cry1Ca5
CAA65457
Strizhov
1996
Bt aizawai 7.29


Cry1Ca6
AAF37224
Yu et al
2000
Bt AF-2


Cry1Ca7
AAG50438
Aixing et al
2000
Bt J8


Cry1Ca8
AAM00264
Chen et al
2001
Bt c002


Cry1Ca9
AAL79362
Kao et al
2003
Bt G10-01A


Cry1Ca10
AAN16462
Lin et al
2003
Bt E05-20a


Cry1Ca11
AAX53094
Cai et al
2005
Bt C-33


Cry1Cb1
M97880
Kalman et al
1993
Bt galleriae HD29
DNA sequence only


Cry1Cb2
AAG35409
Song et al
2000
Bt c001


Cry1Cb3
ACD50894
Huang et al
2008
Bt 087


Cry1Cb-
AAX63901
Thammasittirong et
2005
Bt TA476-1
insufficient sequence


like

al


Cry1Da1
CAA38099
Hofte et al
1990
Bt aizawai HD68


Cry1Da2
I76415
Payne & Sick
1997

DNA sequence only


Cry1Db1
CAA80234
Lambert
1993
Bt BTS00349A


Cry1Db2
AAK48937
Li et al
2001
Bt B-Pr-88


Cry1Dc1
ABK35074
Lertwiriyawong et al
2006
Bt JC291


Cry1Ea1
CAA37933
Visser et al
1990
Bt kenyae 4F1


Cry1Ea2
CAA39609
Bosse et al
1990
Bt kenyae


Cry1Ea3
AAA22345
Feitelson
1991
Bt kenyae PS81F


Cry1Ea4
AAD04732
Barboza-Corona et
1998
Bt kenyae LBIT-




al

147


Cry1Ea5
A15535
Botterman et al
1994

DNA sequence only


Cry1Ea6
AAL50330
Sun et al
1999
Bt YBT-032


Cry1Ea7
AAW72936
Huehne et al
2005
Bt JC190


Cry1Ea8
ABX11258
Huang et al
2007
Bt HZM2


Cry1Eb1
AAA22346
Feitelson
1993
Bt aizawai






PS81A2


Cry1Fa1
AAA22348
Chambers et al
1991
Bt aizawai






EG6346


Cry1Fa2
AAA22347
Feitelson
1993
Bt aizawai PS81I


Cry1Fb1
CAA80235
Lambert
1993
Bt BTS00349A


Cry1Fb2
BAA25298
Masuda & Asano
1998
Bt morrisoni






INA67


Cry1Fb3
AAF21767
Song et al
1998
Bt morrisoni


Cry1Fb4
AAC10641
Payne et al
1997


Cry1Fb5
AAO13295
Li et al
2001
Bt B-Pr-88


Cry1Fb6
ACD50892
Huang et al
2008
Bt 012


Cry1Fb7
ACD50893
Huang et al
2008
Bt 087


Cry1Ga1
CAA80233
Lambert
1993
Bt BTS0349A


Cry1Ga2
CAA70506
Shevelev et al
1997
Bt wuhanensis


Cry1Gb1
AAD10291
Kuo & Chak
1999
Bt wuhanensis






HD525


Cry1Gb2
AAO13756
Li et al
2000
Bt B-Pr-88


Cry1Gc
AAQ52381
Baum et al
2003


Cry1Ha1
CAA80236
Lambert
1993
Bt BTS02069AA


Cry1Hb1
AAA79694
Koo et al
1995
Bt morrisoni






BF190


Cry1H-
AAF01213
Srifah et al
1999
Bt JC291
insufficient sequence


like


Cry1Ia1
CAA44633
Tailor et al
1992
Bt kurstaki


Cry1Ia2
AAA22354
Gleave et al
1993
Bt kurstaki


Cry1Ia3
AAC36999
Shin et al
1995
Bt kurstaki HD1


Cry1Ia4
AAB00958
Kostichka et al
1996
Bt AB88


Cry1Ia5
CAA70124
Selvapandiyan
1996
Bt 61


Cry1Ia6
AAC26910
Zhong et al
1998
Bt kurstaki S101


Cry1Ia7
AAM73516
Porcar et al
2000
Bt


Cry1Ia8
AAK66742
Song et al
2001


Cry1Ia9
AAQ08616
Yao et al
2002
Bt Ly30


Cry1Ia10
AAP86782
Espindola et al
2003
Bt thuringiensis


Cry1Ia11
CAC85964
Tounsi et al
2003
Bt kurstaki BNS3


Cry1Ia12
AAV53390
Grossi de Sa et al
2005
Bt


Cry1Ia13
ABF83202
Martins et al
2006
Bt


Cry1Ia14
ACG63871
Liu & Guo
2008
Bt11


Cry1Ia15
FJ617445
Guan Peng et al
2009
Bt E-1B
No NCBI link July







2009


Cry1Ia16
FJ617448
Guan Peng et al
2009
Bt E-1A
No NCBI link July







2009


Cry1Ib1
AAA82114
Shin et al
1995
Bt entomocidus






BP465


Cry1Ib2
ABW88019
Guan et al
2007
Bt PP61


Cry1Ib3
ACD75515
Liu & Guo
2008
Bt GS8


Cry1Ic1
AAC62933
Osman et al
1998
Bt C18


Cry1Ic2
AAE71691
Osman et al
2001


Cry1Id1
AAD44366
Choi
2000


Cry1Ie1
AAG43526
Song et al
2000
Bt BTC007


Cry1If1
AAQ52382
Baum et al
2003


Cry1I-like
AAC31094
Payne et al
1998

insufficient sequence


Cry1I-like
ABG88859
Lin & Fang
2006
Bt ly4a3
insufficient sequence


Cry1Ja1
AAA22341
Donovan
1994
Bt EG5847


Cry1Jb1
AAA98959
Von Tersch &
1994
Bt EG5092




Gonzalez


Cry1Jc1
AAC31092
Payne et al
1998


Cry1Jc2
AAQ52372
Baum et al
2003


Cry1Jd1
CAC50779
Arnaut et al
2001
Bt


Cry1Ka1
AAB00376
Koo et al
1995
Bt morrisoni






BF190


Cry1La1
AAS60191
Je et al
2004
Bt kurstaki K1


Cry1-like
AAC31091
Payne et al
1998

insufficient sequence


Cry2Aa1
AAA22335
Donovan et al
1989
Bt kurstaki


Cry2Aa2
AAA83516
Widner & Whiteley
1989
Bt kurstaki HD1


Cry2Aa3
D86064
Sasaki et al
1997
Bt sotto
DNA sequence only


Cry2Aa4
AAC04867
Misra et al
1998
Bt kenyae HD549


Cry2Aa5
CAA10671
Yu & Pang
1999
Bt SL39


Cry2Aa6
CAA10672
Yu & Pang
1999
Bt YZ71


Cry2Aa7
CAA10670
Yu & Pang
1999
Bt CY29


Cry2Aa8
AAO13734
Wei et al
2000
Bt Dongbei 66


Cry2Aa9
AAO13750
Zhang et al
2000


Cry2Aa10
AAQ04263
Yao et al
2001


Cry2Aa11
AAQ52384
Baum et al
2003


Cry2Aa12
ABI83671
Tan et al
2006
Bt Rpp39


Cry2Aa13
ABL01536
Arango et al
2008
Bt 146-158-01


Cry2Aa14
ACF04939
Hire et al
2008
Bt HD-550


Cry2Ab1
AAA22342
Widner & Whiteley
1989
Bt kurstaki HD1


Cry2Ab2
CAA39075
Dankocsik et al
1990
Bt kurstaki HD1


Cry2Ab3
AAG36762
Chen et al
1999
Bt BTC002


Cry2Ab4
AAO13296
Li et al
2001
Bt B-Pr-88


Cry2Ab5
AAQ04609
Yao et al
2001
Bt ly30


Cry2Ab6
AAP59457
Wang et al
2003
Bt WZ-7


Cry2Ab7
AAZ66347
Udayasuriyan et al
2005
Bt 14-1


Cry2Ab8
ABC95996
Huang et al
2006
Bt WB2


Cry2Ab9
ABC74968
Zhang et al
2005
Bt LLB6


Cry2Ab10
EF157306
Lin et al
2006
Bt LyD


Cry2Ab11
CAM84575
Saleem et al
2007
Bt CMBL-BT1


Cry2Ab12
ABM21764
Lin et al
2007
Bt LyD


Cry2Ab13
ACG76120
Zhu et al
2008
Bt ywc5-4


Cry2Ab14
ACG76121
Zhu et al
2008
Bt Bts


Cry2Ac1
CAA40536
Aronson
1991
Bt shanghai S1


Cry2Ac2
AAG35410
Song et al
2000


Cry2Ac3
AAQ52385
Baum et al
2003


Cry2Ac4
ABC95997
Huang et al
2006
Bt WB9


Cry2Ac5
ABC74969
Zhang et al
2005


Cry2Ac6
ABC74793
Xia et al
2006
Bt wuhanensis


Cry2Ac7
CAL18690
Saleem et al
2008
Bt SBSBT-1


Cry2Ac8
CAM09325
Saleem et al
2007
Bt CMBL-BT1


Cry2Ac9
CAM09326
Saleem et al
2007
Bt CMBL-BT2


Cry2Ac10
ABN15104
Bai et al
2007
Bt QCL-1


Cry2Ac11
CAM83895
Saleem et al
2007
Bt HD29


Cry2Ac12
CAM83896
Saleem et al
2007
Bt CMBL-BT3


Cry2Ad1
AAF09583
Choi et al
1999
Bt BR30


Cry2Ad2
ABC86927
Huang et al
2006
Bt WB10


Cry2Ad3
CAK29504
Saleem et al
2006
Bt 5_2AcT(1)


Cry2Ad4
CAM32331
Saleem et al
2007
Bt CMBL-BT2


Cry2Ad5
CAO78739
Saleem et al
2007
Bt HD29


Cry2Ae1
AAQ52362
Baum et al
2003


Cry2Af1
ABO30519
Beard et al
2007
Bt C81


Cry2Ag
ACH91610
Zhu et al
2008
Bt JF19-2


Cry2Ah
EU939453
Zhang et al
2008
Bt
No NCBI link July 09


Cry2Ah2
ACL80665
Zhang et al
2009
Bt BRC-ZQL3


Cry2Ai
FJ788388
Udayasuriyan et al
2009
Bt
No NCBI link July 09


Cry3Aa1
AAA22336
Herrnstadt et al
1987
Bt san diego


Cry3Aa2
AAA22541
Sekar et al
1987
Bt tenebrionis


Cry3Aa3
CAA68482
Hofte et al
1987


Cry3Aa4
AAA22542
McPherson et al
1988
Bt tenebrionis


Cry3Aa5
AAA50255
Donovan et al
1988
Bt morrisoni






EG2158


Cry3Aa6
AAC43266
Adams et al
1994
Bt tenebrionis


Cry3Aa7
CAB41411
Zhang et al
1999
Bt 22


Cry3Aa8
AAS79487
Gao and Cai
2004
Bt YM-03


Cry3Aa9
AAW05659
Bulla and Candas
2004
Bt UTD-001


Cry3Aa10
AAU29411
Chen et al
2004
Bt 886


Cry3Aa11
AAW82872
Kurt et al
2005
Bt tenebrionis






Mm2


Cry3Aa12
ABY49136
Sezen et al
2008
Bt tenebrionis


Cry3Ba1
CAA34983
Sick et al
1990
Bt tolworthi 43F


Cry3Ba2
CAA00645
Peferoen et al
1990
Bt PGSI208


Cry3Bb1
AAA22334
Donovan et al
1992
Bt EG4961


Cry3Bb2
AAA74198
Donovan et al
1995
Bt EG5144


Cry3Bb3
I15475
Peferoen et al
1995

DNA sequence only


Cry3Ca1
CAA42469
Lambert et al
1992
Bt kurstaki






BtI109P


Cry4Aa1
CAA68485
Ward & Ellar
1987
Bt israelensis


Cry4Aa2
BAA00179
Sen et al
1988
Bt israelensis






HD522


Cry4Aa3
CAD30148
Berry et al
2002
Bt israelensis


Cry4A-
AAY96321
Mahalakshmi et al
2005
Bt LDC-9
insufficient sequence


like


Cry4Ba1
CAA30312
Chungjatpornchai et
1988
Bt israelensis




al

4Q2-72


Cry4Ba2
CAA30114
Tungpradubkul et al
1988
Bt israelensis


Cry4Ba3
AAA22337
Yamamoto et al
1988
Bt israelensis


Cry4Ba4
BAA00178
Sen et al
1988
Bt israelensis






HD522


Cry4Ba5
CAD30095
Berry et al
2002
Bt israelensis


Cry4Ba-
ABC47686
Mahalakshmi et al
2005
Bt LDC-9
insufficient sequence


like


Cry4Ca1
EU646202
Shu et al
2008

No NCBI link July 09


Cry4Cb1
FJ403208
Jun & Furong
2008
Bt HS18-1
No NCBI link July 09


Cry4Cb2
FJ597622
Jun & Furong
2008
Bt Ywc2-8
No NCBI link July 09


Cry4Cc1
FJ403207
Jun & Furong
2008
Bt MC28
No NCBI link July 09


Cry5Aa1
AAA67694
Narva et al
1994
Bt darmstadiensis






PS17


Cry5Ab1
AAA67693
Narva et al
1991
Bt darmstadiensis






PS17


Cry5Ac1
I34543
Payne et al
1997

DNA sequence only


Cry5Ad1
ABQ82087
Lenane et al
2007
Bt L366


Cry5Ba1
AAA68598
Foncerrada & Narva
1997
Bt PS86Q3


Cry5Ba2
ABW88932
Guo et al
2008
YBT 1518


Cry6Aa1
AAA22357
Narva et al
1993
Bt PS52A1


Cry6Aa2
AAM46849
Bai et al
2001
YBT 1518


Cry6Aa3
ABH03377
Jia et al
2006
Bt 96418


Cry6Ba1
AAA22358
Narva et al
1991
Bt PS69D1


Cry7Aa1
AAA22351
Lambert et al
1992
Bt galleriae






PGSI245


Cry7Ab1
AAA21120
Narva & Fu
1994
Bt dakota HD511


Cry7Ab2
AAA21121
Narva & Fu
1994
Bt kumamotoensis






867


Cry7Ab3
ABX24522
Song et al
2008
Bt WZ-9


Cry7Ab4
EU380678
Shu et al
2008
Bt
No NCBI link July 09


Cry7Ab5
ABX79555
Aguirre-Arzola et al
2008
Bt monterrey GM-






33


Cry7Ab6
ACI44005
Deng et al
2008
Bt HQ122


Cry7Ab7
FJ940776
Wang et al
2009

No NCBI link Sept 09


Cry7Ab8
GU145299
Feng Jing
2009

No NCBI link Nov 09


Cry7Ba1
ABB70817
Zhang et al
2006
Bt huazhongensis


Cry7Ca1
ABR67863
Gao et al
2007
Bt BTH-13


Cry7Da1
ACQ99547
Yi et al
2009
Bt LH-2


Cry8Aa1
AAA21117
Narva & Fu
1992
Bt kumamotoensis


Cry8Ab1
EU044830
Cheng et al
2007
Bt B-JJX
No NCBI link July 09


Cry8Ba1
AAA21118
Narva & Fu
1993
Bt kumamotoensis


Cry8Bb1
CAD57542
Abad et al
2002


Cry8Bc1
CAD57543
Abad et al
2002


Cry8Ca1
AAA21119
Sato et al.
1995
Bt japonensis






Buibui


Cry8Ca2
AAR98783
Shu et al
2004
Bt HBF-1


Cry8Ca3
EU625349
Du et al
2008
Bt FTL-23
No NCBI link July 09


Cry8Da1
BAC07226
Asano et al
2002
Bt galleriae


Cry8Da2
BD133574
Asano et al
2002
Bt
DNA sequence only


Cry8Da3
BD133575
Asano et al
2002
Bt
DNA sequence only


Cry8Db1
BAF93483
Yamaguchi et al
2007
Bt BBT2-5


Cry8Ea1
AAQ73470
Fuping et al
2003
Bt 185


Cry8Ea2
EU047597
Liu et al
2007
Bt B-DLL
No NCBI link July 09


Cry8Fa1
AAT48690
Shu et al
2004
Bt 185
also AAW81032


Cry8Ga1
AAT46073
Shu et al
2004
Bt HBF-18


Cry8Ga2
ABC42043
Yan et al
2008
Bt 145


Cry8Ga3
FJ198072
Xiaodong et al
2008
Bt FCD114
No NCBI link July 09


Cry8Ha1
EF465532
Fuping et al
2006
Bt 185
No NCBI link July 09


Cry8Ia1
EU381044
Yan et al
2008
Bt su4
No NCBI link July 09


Cry8Ja1
EU625348
Du et al
2008
Bt FPT-2
No NCBI link July 09


Cry8Ka1
FJ422558
Quezado et al
2008

No NCBI link July 09


Cry8Ka2
ACN87262
Noguera & Ibarra
2009
Bt kenyae


Cry8-like
FJ770571
Noguera & Ibarra
2009
Bt canadensis
DNA sequence only


Cry8-like
ABS53003
Mangena et al
2007
Bt


Cry9Aa1
CAA41122
Shevelev et al
1991
Bt galleriae


Cry9Aa2
CAA41425
Gleave et al
1992
Bt DSIR517


Cry9Aa3
GQ249293
Su et al
2009
Bt SC5(D2)
No NCBI link July 09


Cry9Aa4
GQ249294
Su et al
2009
Bt T03C001
No NCBI link July 09


Cry9Aa
AAQ52376
Baum et al
2003

incomplete sequence


like


Cry9Ba1
CAA52927
Shevelev et al
1993
Bt galleriae


Cry9Bb1
AAV28716
Silva-Werneck et al
2004
Bt japonensis


Cry9Ca1
CAA85764
Lambert et al
1996
Bt tolworthi


Cry9Ca2
AAQ52375
Baum et al
2003


Cry9Da1
BAA19948
Asano
1997
Bt japonensis






N141


Cry9Da2
AAB97923
Wasano & Ohba
1998
Bt japonensis


Cry9Da3
GQ249295
Su et al
2009
Bt T03B001
No NCBI link July 09


Cry9Da4
GQ249297
Su et al
2009
Bt T03B001
No NCBI link July 09


Cry9Db1
AAX78439
Flannagan & Abad
2005
Bt kurstaki






DP1019


Cry9Ea1
BAA34908
Midoh & Oyama
1998
Bt aizawai SSK-






10


Cry9Ea2
AAO12908
Li et al
2001
Bt B-Hm-16


Cry9Ea3
ABM21765
Lin et al
2006
Bt lyA


Cry9Ea4
ACE88267
Zhu et al
2008
Bt ywc5-4


Cry9Ea5
ACF04743
Zhu et al
2008
Bts


Cry9Ea6
ACG63872
Liu & Guo
2008
Bt 11


Cry9Ea7
FJ380927
Sun et al
2008

No NCBI link July 09


Cry9Ea8
GQ249292
Su et al
2009
GQ249292
No NCBI link July 09


Cry9Eb1
CAC50780
Arnaut et al
2001


Cry9Eb2
GQ249298
Su et al
2009
Bt T03B001
No NCBI link July 09


Cry9Ec1
AAC63366
Wasano et al
2003
Bt galleriae


Cry9Ed1
AAX78440
Flannagan & Abad
2005
Bt kurstaki






DP1019


Cry9Ee1
GQ249296
Su et al
2009
Bt T03B001
No NCBI link Aug 09


Cry9-like
AAC63366
Wasano et al
1998
Bt galleriae
insufficient sequence


Cry10Aa1
AAA22614
Thorne et al
1986
Bt israelensis


Cry10Aa2
E00614
Aran & Toomasu
1996
Bt israelensis
DNA sequence only






ONR-60A


Cry10Aa3
CAD30098
Berry et al
2002
Bt israelensis


Cry10A-
DQ167578
Mahalakshmi et al
2006
Bt LDC-9
incomplete sequence


like


Cry11Aa1
AAA22352
Donovan et al
1988
Bt israelensis


Cry11Aa2
AAA22611
Adams et al
1989
Bt israelensis


Cry11Aa3
CAD30081
Berry et al
2002
Bt israelensis


Cry11Aa-
DQ166531
Mahalakshmi et al
2007
Bt LDC-9
incomplete sequence


like


Cry11Ba1
CAA60504
Delecluse et al
1995
Bt jegathesan 367


Cry11Bb1
AAC97162
Orduz et al
1998
Bt medellin


Cry12Aa1
AAA22355
Narva et al
1991
Bt PS33F2


Cry13Aa1
AAA22356
Narva et al
1992
Bt PS63B


Cry14Aa1
AAA21516
Narva et al
1994
Bt sotto PS80JJ1


Cry15Aa1
AAA22333
Brown & Whiteley
1992
Bt thompsoni


Cry16Aa1
CAA63860
Barloy et al
1996
Cb malaysia CH18


Cry17Aa1
CAA67841
Barloy et al
1998
Cb malaysia CH18


Cry18Aa1
CAA67506
Zhang et al
1997
Paenibacillus






popilliae


Cry18Ba1
AAF89667
Patel et al
1999
Paenibacillus






popilliae


Cry18Ca1
AAF89668
Patel et al
1999
Paenibacillus






popilliae


Cry19Aa1
CAA68875
Rosso & Delecluse
1996
Bt jegathesan 367


Cry19Ba1
BAA32397
Hwang et al
1998
Bt higo


Cry20Aa1
AAB93476
Lee & Gill
1997
Bt fukuokaensis


Cry20Ba1
ACS93601
Noguera & Ibarra
2009
Bt higo LBIT-976


Cry20-like
GQ144333
Yi et al
2009
Bt Y-5
DNA sequence only


Cry21Aa1
I32932
Payne et al
1996

DNA sequence only


Cry21Aa2
I66477
Feitelson
1997

DNA sequence only


Cry21Ba1
BAC06484
Sato & Asano
2002
Bt roskildiensis


Cry22Aa1
I34547
Payne et al
1997

DNA sequence only


Cry22Aa2
CAD43579
Isaac et al
2002
Bt


Cry22Aa3
ACD93211
Du et al
2008
Bt FZ-4


Cry22Ab1
AAK50456
Baum et al
2000
Bt EG4140


Cry22Ab2
CAD43577
Isaac et al
2002
Bt


Cry22Ba1
CAD43578
Isaac et al
2002
Bt


Cry23Aa1
AAF76375
Donovan et al
2000
Bt
Binary with Cry37Aa1


Cry24Aa1
AAC61891
Kawalek and Gill
1998
Bt jegathesan


Cry24Ba1
BAD32657
Ohgushi et al
2004
Bt sotto


Cry24Ca1
CAJ43600
Beron & Salerno
2005
Bt FCC-41


Cry25Aa1
AAC61892
Kawalek and Gill
1998
Bt jegathesan


Cry26Aa1
AAD25075
Wojciechowska et
1999
Bt finitimus B-




al

1166


Cry27Aa1
BAA82796
Saitoh
1999
Bt higo


Cry28Aa1
AAD24189
Wojciechowska et al
1999
Bt finitimus B-






1161


Cry28Aa2
AAG00235
Moore and Debro
2000
Bt finitimus


Cry29Aa1
CAC80985
Delecluse et al
2000
Bt medellin


Cry30Aa1
CAC80986
Delecluse et al
2000
Bt medellin


Cry30Ba1
BAD00052
Ito et al
2003
Bt entomocidus


Cry30Ca1
BAD67157
Ohgushi et al
2004
Bt sotto


Cry30Ca2
ACU24781
Sun and Park
2009
Bt jegathesan 367


Cry30Da1
EF095955
Shu et al
2006
Bt Y41
No NCBI link July09


Cry30Db1
BAE80088
Kishida et al
2006
Bt aizawai BUN1-






14


Cry30Ea1
ACC95445
Fang et al
2007
Bt S2160-1


Cry30Ea2
FJ499389
Jun et al
2008
Bt Ywc2-8
No NCBI link July09


Cry30Fa1
ACI22625
Tan et al
2008
Bt MC28


Cry30Ga1
ACG60020
Zhu et al
2008
Bt HS18-1


Cry31Aa1
BAB11757
Saitoh & Mizuki
2000
Bt 84-HS-1-11


Cry31Aa2
AAL87458
Jung and Cote
2000
Bt M15


Cry31Aa3
BAE79808
Uemori et al
2006
Bt B0195


Cry31Aa4
BAF32571
Yasutake et al
2006
Bt 79-25


Cry31Aa5
BAF32572
Yasutake et al
2006
Bt 92-10


Cry31Ab1
BAE79809
Uemori et al
2006
Bt B0195


Cry31Ab2
BAF32570
Yasutake et al
2006
Bt 31-5


Cry31Ac1
BAF34368
Yasutake et al
2006
Bt 87-29


Cry32Aa1
AAG36711
Balasubramanian et
2001
Bt yunnanensis




al


Cry32Ba1
BAB78601
Takebe et al
2001
Bt


Cry32Ca1
BAB78602
Takebe et al
2001
Bt


Cry32Da1
BAB78603
Takebe et al
2001
Bt


Cry33Aa1
AAL26871
Kim et al
2001
Bt dakota


Cry34Aa1
AAG50341
Ellis et al
2001
Bt PS80JJ1
Binary with Cry35Aa1


Cry34Aa2
AAK64560
Rupar et al
2001
Bt EG5899
Binary with Cry35Aa2


Cry34Aa3
AAT29032
Schnepf et al
2004
Bt PS69Q
Binary with Cry35Aa3


Cry34Aa4
AAT29030
Schnepf et al
2004
Bt PS185GG
Binary with Cry35Aa4


Cry34Ab1
AAG41671
Moellenbeck et al
2001
Bt PS149B1
Binary with Cry35Ab1


Cry34Ac1
AAG50118
Ellis et al
2001
Bt PS167H2
Binary with Cry35Ac1


Cry34Ac2
AAK64562
Rupar et al
2001
Bt EG9444
Binary with Cry35Ab2


Cry34Ac3
AAT29029
Schnepf et al
2004
Bt KR1369
Binary with Cry35Ab3


Cry34Ba1
AAK64565
Rupar et al
2001
Bt EG4851
Binary with Cry35Ba1


Cry34Ba2
AAT29033
Schnepf et al
2004
Bt PS201L3
Binary with Cry35Ba2


Cry34Ba3
AAT29031
Schnepf et al
2004
Bt PS201HH2
Binary with Cry35Ba3


Cry35Aa1
AAG50342
Ellis et al
2001
Bt PS80111
Binary with Cry34Aa1


Cry35Aa2
AAK64561
Rupar et al
2001
Bt EG5899
Binary with Cry34Aa2


Cry35Aa3
AAT29028
Schnepf et al
2004
Bt PS69Q
Binary with Cry34Aa3


Cry35Aa4
AAT29025
Schnepf et al
2004
Bt PS185GG
Binary with Cry34Aa4


Cry35Ab1
AAG41672
Moellenbeck et al
2001
Bt PS149B1
Binary with Cry34Ab1


Cry35Ab2
AAK64563
Rupar et al
2001
Bt EG9444
Binary with Cry34Ac2


Cry35Ab3
AY536891
AAT29024
2004
Bt KR1369
Binary with Cry34Ab3


Cry35Ac1
AAG50117
Ellis et al
2001
Bt PS167H2
Binary with Cry34Ac1


Cry35Ba1
AAK64566
Rupar et al
2001
Bt EG4851
Binary with Cry34Ba1


Cry35Ba2
AAT29027
Schnepf et al
2004
Bt PS201L3
Binary with Cry34Ba2


Cry35Ba3
AAT29026
Schnepf et al
2004
Bt PS201HH2
Binary with Cry34Ba3


Cry36Aa1
AAK64558
Rupar et al
2001
Bt


Cry37Aa1
AAF76376
Donovan et al
2000
Bt
Binary with Cry23Aa


Cry38Aa1
AAK64559
Rupar et al
2000
Bt


Cry39Aa1
BAB72016
Ito et al
2001
Bt aizawai


Cry40Aa1
BAB72018
Ito et al
2001
Bt aizawai


Cry40Ba1
BAC77648
Ito et al
2003
Bun1-14


Cry40Ca1
EU381045
Shu et al
2008
Bt Y41
No NCBI link July09


Cry40Da1
ACF15199
Zhang et al
2008
Bt S2096-2


Cry41Aa1
BAD35157
Yamashita et al
2003
Bt A1462


Cry41Ab1
BAD35163
Yamashita et al
2003
Bt A1462


Cry42Aa1
BAD35166
Yamashita et al
2003
Bt A1462


Cry43Aa1
BAD15301
Yokoyama and
2003

P. lentimorbus





Tanaka


semadara



Cry43Aa2
BAD95474
Nozawa
2004

P. popilliae








popilliae



Cry43Ba1
BAD15303
Yokoyama and
2003

P. lentimorbus





Tanaka


semadara



Cry43-like
BAD15305
Yokoyama and
2003

P. lentimorbus





Tanaka


semadara



Cry44Aa
BAD08532
Ito et al
2004
Bt entomocidus






INA288


Cry45Aa
BAD22577
Okumura et al
2004
Bt 89-T-34-22


Cry46Aa
BAC79010
Ito et al
2004
Bt dakota


Cry46Aa2
BAG68906
Ishikawa et al
2008
Bt A1470


Cry46Ab
BAD35170
Yamagiwa et al
2004
Bt


Cry47Aa
AAY24695
Kongsuwan et al
2005
Bt CAA890


Cry48Aa
CAJ18351
Jones and Berry
2005
Bs IAB59
binary with 49Aa


Cry48Aa2
CAJ86545
Jones and Berry
2006
Bs 47-6B
binary with 49Aa2


Cry48Aa3
CAJ86546
Jones and Berry
2006
Bs NHA15b
binary with 49Aa3


Cry48Ab
CAJ86548
Jones and Berry
2006
Bs LP1G
binary with 49Ab1


Cry48Ab2
CAJ86549
Jones and Berry
2006
Bs 2173
binary with 49Aa4


Cry49Aa
CAH56541
Jones and Berry
2005
Bs IAB59
binary with 48Aa


Cry49Aa2
CAJ86541
Jones and Berry
2006
Bs 47-6B
binary with 48Aa2


Cry49Aa3
CAJ86543
Jones and Berry
2006
BsNHA15b
binary with 48Aa3


Cry49Aa4
CAJ86544
Jones and Berry
2006
Bs 2173
binary with 48Ab2


Cry49Ab1
CAJ86542
Jones and Berry
2006
Bs LP1G
binary with 48Ab1


Cry50Aa1
BAE86999
Ohgushi et al
2006
Bt sotto


Cry51Aa1
ABI14444
Meng et al
2006
Bt F14-1


Cry52Aa1
EF613489
Song et al
2007
Bt Y41
No NCBI link July09


Cry52Ba1
FJ361760
Jun et al
2008
Bt BM59-2
No NCBI link July09


Cry53Aa1
EF633476
Song et al
2007
Bt Y41
No NCBI link July09


Cry53Ab1
FJ361759
Jun et al
2008
Bt MC28
No NCBI link July09


Cry54Aa1
ACA52194
Tan et al
2009
Bt MC28


Cry55Aa1
ABW88931
Guo et al
2008
YBT 1518


Cry55Aa2
AAE33526
Bradfisch et al
2000
BT Y41


Cry56Aa1
FJ597621
Jun & Furong
2008
Bt Ywc2-8
No NCBI link July09


Cry56Aa2
GQ483512
Guan Peng et al
2009
Bt G7-1
No NCBI link Aug09


Cry57Aa1
ANC87261
Noguera & Ibarra
2009
Bt kim


Cry58Aa1
ANC87260
Noguera & Ibarra
2009
Bt entomocidus


Cry59Aa1
ACR43758
Noguera & Ibarra
2009
Bt kim LBIT-980




























Vip3Aa1
Vip3Aa
AAC37036
Estruch et al
1996
PNAS 93,
AB88








5389-5394


Vip3Aa2
Vip3Ab
AAC37037
Estruch et al
1996
PNAS 93,
AB424







5389-5394


Vip3Aa3
Vip3Ac

Estruch et al
2000
U.S. Pat. No.







6,137,033







October 2000


Vip3Aa4
PS36A Sup
AAR81079
Feitelson et al
1998
U.S. Pat. No.
Bt PS36A
WO9818932(A2, A3)







6,656,908

7 May 1998







December 2003


Vip3Aa5
PS81F Sup
AAR81080
Feitelson et al
1998
U.S. Pat. No.
Bt PS81F
WO9818932(A2, A3)







6,656,908

7 May 1998







December 2003


Vip3Aa6
Jav90 Sup
AAR81081
Feitelson et al
1998
U.S. Pat. No.
Bt
WO9818932(A2, A3)







6,656,908

7 May 1998







December 2003


Vip3Aa7
Vip83
AAK95326
Cai et al
2001
unpublished
Bt YBT-833


Vip3Aa8
Vip3A
AAK97481
Loguercio et al
2001
unpublished
Bt HD125


Vip3Aa9
VipS
CAA76665
Selvapandiyan
2001
unpublished
Bt A13





et al


Vip3Aa10
Vip3V
AAN60738
Doss et al
2002
Protein Expr.
Bt







Purif. 26, 82-88


Vip3Aa11
Vip3A
AAR36859
Liu et al
2003
unpublished
Bt C9


Vip3Aa12
Vip3A-WB5
AAM22456
Wu and Guan
2003
unpublished
Bt


Vip3Aa13
Vip3A
AAL69542
Chen et al
2002
Sheng Wu
Bt S184







Gong Cheng







Xue Bao 18,







687-692


Vip3Aa14
Vip
AAQ12340
Polumetla et al
2003
unpublished
Bt tolworthi


Vip3Aa15
Vip3A
AAP51131
Wu et al
2004
unpublished
Bt WB50


Vip3Aa16
Vip3LB
AAW65132
Mesrati et al
2005
FEMS Micro
Bt







Lett 244,







353-358


Vip3Aa17
Jav90

Feitelson et al
1999
U.S. Pat. No.
Javelin 1990
WO9957282(A2, A3)







6,603,063

11 Nov. 1999







August 2003


Vip3Aa18

AAX49395
Cai and Xiao
2005
unpublished
Bt 9816C


Vip3Aa19
Vip3ALD
DQ241674
Liu et al
2006
unpublished
Bt AL


Vip3Aa19
Vip3A-1
DQ539887
Hart et al
2006
unpublished


Vip3Aa20
Vip3A-2
DQ539888
Hart et al
2006
unpublished


Vip3Aa21
Vip
ABD84410
Panbangred
2006
unpublished
Bt aizawai


Vip3Aa22
Vip3A-LS1
AAY41427
Lu et al
2005
unpublished
Bt LS1


Vip3Aa23
Vip3A-LS8
AAY43428
Lu et al
2005
unpublished
Bt LS8


Vip3Aa24

BI 880913
Song et al
2007
unpublished
Bt WZ-7


Vip3Aa25

EF608501
Hsieh et al
2007
unpublished


Vip3Aa26

EU294496
Shen and Guo
2007
unpublished
Bt TF9


Vip3Aa27

EU332167
Shen and Guo
2007
unpublished
Bt 16


Vip3Aa28

FJ494817
Xiumei Yu
2008
unpublished
Bt JF23-8


Vip3Aa29

FJ626674
Xieumei et al
2009
unpublished
Bt JF21-1


Vip3Aa30

FJ626675
Xieumei et al
2009
unpublished
MD2-1


Vip3Aa31

FJ626676
Xieumei et al
2009
unpublished
JF21-1


Vip3Aa32

FJ626677
Xieumei et al
2009
unpublished
MD2-1


.



.


Vip3Ab1
Vip3B
AAR40284
Feitelson et al
1999
U.S. Pat. No.
Bt KB59A4-6
WO9957282(A2, A3)







6,603,063

11 Nov. 1999







August 2003


Vip3Ab2
Vip3D
AAY88247
Feng and Shen
2006
unpublished
Bt


.



.


Vip3Ac1
PS49C

Narva et al
.
US application







20040128716


.



.


Vip3Ad1
PS158C2

Narva et al
.
US application







20040128716


Vip3Ad2
ISP3B
CAI43276
Van Rie et al
2005
unpublished
Bt


.



.


Vip3Ae1
ISP3C
CAI43277
Van Rie et al
2005
unpublished
Bt


.



.


Vip3Af1
ISP3A
CAI43275
Van Rie et al
2005
unpublished
Bt


Vip3Af2
Vip3C
ADN08753
Syngenta
.
WO 03/075655


.



.


Vip3Ag1
Vip3B
ADN08758
Syngenta
.
WO 02/078437


Vip3Ag2

FJ556803
Audtho et al
2008

Bt


.



.


Vip3Ah1
Vip3S
DQ832323
Li and Shen
2006
unpublished
Bt


.


Vip3Ba1

AAV70653
Rang et al
2004
unpublished


.


Vip3Bb1
Vip3Z
ADN08760
Syngenta
.
WO 03/075655


Vip3Bb2

EF439819
Akhurst et al
2007








Claims
  • 1. A transgenic plant comprising DNA encoding a Cry1Fa insecticidal protein and DNA encoding a second insecticidal protein selected from the group consisting of a Cry2Aa insecticidal protein, a Cry1I insecticidal protein, and a Cry1E insecticidal protein.
  • 2. (canceled)
  • 3. The transgenic plant of claim 1, said plant further comprising DNA encoding a third insecticidal protein, said third protein being selected from the group consisting of Cry1Be, Cry1Ab, and DIG-3.
  • 4. The transgenic plant of claim 3, wherein said third protein is selected from the group consisting of Cry1Be, said plant further comprising DNA encoding fourth insecticidal protein selected from the group consisting of Cry1Ca, Cry1Da, and Vip3Ab.
  • 5. The transgenic plant of claim 1, wherein said second protein is a Cry1E insecticidal protein, and said plant further comprising DNA encoding a third insecticidal protein, said third protein being selected from the group consisting of Cry1Be, Cry1Ca, Cry1Da, and Vip3Ab.
  • 6. The transgenic plant of claim 5, wherein said third protein is selected from the group consisting of Cry1Be, said plant further comprising DNA encoding fourth insecticidal protein selected from the group consisting of Cry1Ab, Cry1I, Cry2A, and DIG-3.
  • 7. Seed of a plant according to claim 1, wherein said seed comprises said DNA.
  • 8. A field of plants comprising non-Bt refuge plants and a plurality of plants according to claim 1, wherein said refuge plants comprise less than 40% of all crop plants in said field.
  • 9. (canceled)
  • 10. (canceled)
  • 11. (canceled)
  • 12. (canceled)
  • 13. (canceled)
  • 14. A mixture of seeds comprising refuge seeds from non-Bt refuge plants, and a plurality of seeds of claim 7, wherein said refuge seeds comprise less than 40% of all the seeds in the mixture.
  • 15. (canceled)
  • 16. (canceled)
  • 17. (canceled)
  • 18. (canceled)
  • 19. A method of managing development of resistance to a Cry protein by an insect, said method comprising planting seeds to produce a field of plants of claim 8.
  • 20. (canceled)
  • 21. The plant of claim 1, wherein said plant is selected from the group consisting of corn, soybeans, and cotton.
  • 22. (canceled)
  • 23. A method of controlling an insect by contacting said insect with a Cry1F insecticidal protein and a second insecticidal protein selected from the group consisting of a Cry2Aa insecticidal protein, a Cry1I insecticidal protein, and a Cry1E insecticidal protein.
  • 24. An isolated polynucleotide that encodes a Cry1F insecticidal protein in combination with a second insecticidal protein selected from the group consisting of a Cry2Aa insecticidal protein, and a Cry1I insecticidal protein, a Cry1E insecticidal protein.
  • 25. A method of making a Cry1F insecticidal protein in combination with a second insecticidal protein selected from the group consisting of a Cry2Aa insecticidal protein, a Cry1I insecticidal protein, a Cry1E insecticidal protein.
  • 26. (canceled)
  • 27. (canceled)
  • 28. (canceled)
  • 29. A plant cell of a plant of claim 1, wherein said Cry1Fa insecticidal protein is at least 99% identical with SEQ ID NO:1, said Cry2Aa insecticidal protein is at least 99% identical with SEQ ID NO:2, said Cry1I insecticidal protein is at least 99% identical with SEQ ID NO:3.
  • 30. (canceled)
  • 31. Seed of a plant according to claim 3, wherein said seed comprises said DNA.
  • 32. Seed of a plant according to claim 4, wherein said seed comprises said DNA.
  • 33. Seed of a plant according to claim 5, wherein said seed comprises said DNA.
  • 34. Seed of a plant according to claim 6, wherein said seed comprises said DNA.
  • 35. A mixture of seeds comprising refuge seeds from non-Bt refuge plants, and a plurality of seeds of claim 31, wherein said refuge seeds comprise less than 40% of all the seeds in the mixture.
  • 36. A mixture of seeds comprising refuge seeds from non-Bt refuge plants, and a plurality of seeds of claim 32, wherein said refuge seeds comprise less than 40% of all the seeds in the mixture.
  • 37. A mixture of seeds comprising refuge seeds from non-Bt refuge plants, and a plurality of seeds of claim 33, wherein said refuge seeds comprise less than 40% of all the seeds in the mixture.
  • 38. A mixture of seeds comprising refuge seeds from non-Bt refuge plants, and a plurality of seeds of claim 34, wherein said refuge seeds comprise less than 40% of all the seeds in the mixture.
  • 39. The method of claim 23, wherein the insect is selected from the group consisting of a European cornborer and a fall armyworm.
PCT Information
Filing Document Filing Date Country Kind 371c Date
PCT/US10/60812 12/16/2010 WO 00 8/30/2012
Provisional Applications (1)
Number Date Country
61284278 Dec 2009 US