Insect inhibitory proteins

Patent metadata

Field	Value
Publication number	US-10626151-B2
Application number	US-201715727883-A
Country	US
Kind code	B2
Filing date	Oct 9, 2017
Priority date	Oct 10, 2016
Publication date	Apr 21, 2020
Grant date	Apr 21, 2020

Abstract

Pesticidal proteins exhibiting toxic activity against Lepidopteran pest species are disclosed, and include, but are not limited to, TIC4472, TIC4472PL, TIC1425, TIC2613, and TIC2613PL. DNA constructs are provided which contain a recombinant nucleic acid sequence encoding one or more of the disclosed pesticidal proteins. Transgenic plants, plant cells, seed, and plant parts resistant to Lepidopteran infestation are provided which contain recombinant nucleic acid sequences encoding the pesticidal proteins of the present invention. Methods for detecting the presence of the recombinant nucleic acid sequences or the proteins of the present invention in a biological sample, and methods of controlling Lepidopteran species pests using any of the TIC4472, TIC4472PL, TIC1425, TIC2613, and TIC2613PL pesticidal proteins are also provided.

First claim

What is claimed is:

1. A recombinant nucleic acid molecule comprising a heterologous promoter operably linked to a polynucleotide segment encoding a pesticidal protein or pesticidal fragment thereof, wherein:
a. said pesticidal protein comprises the amino acid sequence of SEQ ID NO:4, SEQ ID NO:2, or SEQ ID NO:6 and exhibits pesticidal activity; or
b. said pesticidal protein comprises an amino acid sequence having
at least 93%, or 95%, or 98% or 99%, or about 100% amino acid sequence identity to SEQ ID NO:4, SEQ ID NO:2, or SEQ ID NO:6 and exhibits pesticidal activity; or

c. said polynucleotide segment hybridizes under stringent hybridization conditions to a polynucleotide having the nucleotide sequence of SEQ ID NO:3, SEQ ID NO:1, or SEQ ID NO:5 and said encoded pesticidal protein exhibits pesticidal activity.

2. The recombinant nucleic acid molecule of claim 1 , wherein:
a. the recombinant nucleic acid molecule comprises a sequence that functions to express the pesticidal protein in a plant; or
b. the recombinant nucleic acid molecule is expressed in a plant cell to produce a pesticidally effective amount of the pesticidal protein; or
c. the recombinant nucleic acid molecule is in operable linkage with a vector, and said vector is selected from the group consisting of a plasmid, phagemid, bacmid, cosmid, and a bacterial or yeast artificial chromosome.

3. A host cell comprising the recombinant nucleic acid molecule of claim 1 , wherein said host cell is selected from the group consisting of a bacterial and a plant cell.

4. The host cell of claim 3 , wherein the bacterial host cell is from a genus of bacteria selected from the group consisting of: Agrobacterium, Rhizobium, Bacillus, Brevibacillus, Escherichia, Pseudomonas, Klebsiella, Pantoea , and Erwinia.

5. The host cell of claim 4 wherein the Bacillus species is Bacillus cereus or Bacillus thuringiensis , said Brevibacillus is Brevibacillus laterosperous , and said Escherichia is Escherichia coli.

6. The host cell of claim 3 , wherein said plant cell is a dicotyledonous or a monocotyledonous plant cell.

7. The host cell of claim 6 , wherein said plant cell is selected from the group consisting of an alfalfa, banana, barley, bean, broccoli, cabbage, brassica, carrot, cassava, castor, cauliflower, celery, chickpea, Chinese cabbage, citrus, coconut, coffee, corn, clover, cotton, a cucurbit, cucumber, Douglas fir, eggplant, eucalyptus, flax, garlic, grape, hops, leek, lettuce, Loblolly pine, millets, melons, nut, oat, olive, onion, ornamental, palm, pasture grass, pea, peanut, pepper, pigeonpea, pine, potato, poplar, pumpkin, Radiata pine, radish, rapeseed, rice, rootstocks, rye, safflower, shrub, sorghum, Southern pine, soybean, spinach, squash, strawberry, sugar beet, sugarcane, sunflower, sweet corn, sweet gum, sweet potato, switchgrass, tea, tobacco, tomato, triticale, turf grass, watermelon, and wheat plant cell.

8. The recombinant nucleic acid molecule of claim 1 , wherein said protein exhibits activity against a Lepidopteran insect.

9. The recombinant nucleic acid molecule of claim 8 , wherein said insect is selected from the group consisting of: Beet armyworm ( Spodoptera exigua ), Corn earworm ( Helicoverpa zea ), Cotton leaf worm ( Alabama argillacea ), European corn borer ( Ostrinia nubilalis ), Fall armyworm ( Spodoptera frugiperda ), Old World bollworm ( Helicoverpa armigera ), Oriental leaf worm ( Spodoptera litura ), Pink bollworm ( Pectinophora gossypiella ), Cry1Ac resistant Pink bollworm ( Pectinophora gossypiella ), Soybean looper ( Chrysodeixis includens ), Southern armyworm ( Spodoptera eridania ), Southwestern corn borer ( Diatraea grandiosella ), Spotted bollworm ( Earias vittella ), Sugarcane borer ( Diatraea saccharalis ), Tobacco budworm ( Heliothis virescens ), and Velvet bean caterpillar ( Anticarsia gemmatalis ).

10. A plant or part thereof comprising the recombinant nucleic acid molecule of claim 1 , wherein said plant is resistant to insect infestation.

11. The plant or part thereof of claim 10 , wherein said plant is a monocot plant or a dicot plant.

12. The plant of claim 10 , wherein the plant is selected from the group consisting of an alfalfa, banana, barley, bean, broccoli, cabbage, brassica, carrot, cassava, castor, cauliflower, celery, chickpea, Chinese cabbage, citrus, coconut, coffee, corn, clover, cotton, a cucurbit, cucumber, Douglas fir, eggplant, eucalyptus, flax, garlic, grape, hops, leek, lettuce, Loblolly pine, millets, melons, nut, oat, olive, onion, ornamental, palm, pasture grass, pea, peanut, pepper, pigeon pea, pine, potato, poplar, pumpkin, Radiata pine, radish, rapeseed, rice, rootstocks, rye, safflower, shrub, sorghum, Southern pine, soybean, spinach, squash, strawberry, sugar beet, sugarcane, sunflower, sweet corn, sweet gum, sweet potato, switchgrass, tea, tobacco, tomato, triticale, turf grass, watermelon, and wheat.

13. A seed of the plant of claim 10 , wherein said seed comprises said recombinant nucleic acid molecule.

14. An insect inhibitory composition comprising the recombinant nucleic acid molecule of claim 1 .

15. The insect inhibitory composition of claim 14 , further comprising a nucleotide sequence encoding at least one other pesticidal agent that is different from said pesticidal protein.

16. The insect inhibitory composition of claim 15 , wherein said at least one other pesticidal agent is selected from the group consisting of an insect inhibitory protein, an insect inhibitory dsRNA molecule, and an ancillary protein.

17. The insect inhibitory composition of claim 16 , wherein said at least one other pesticidal agent exhibits activity against one or more pest species of the orders Lepidoptera, Coleoptera, or Hemiptera.

18. The insect inhibitory composition of claim 17 , wherein said at least one other pesticidal protein is selected from the group consisting of a Cry1A, Cry1Ab, Cry1Ac, Cry1A.105, Cry1Ae, Cry1B, Cry1C, Cry1C variants, Cry1D, Cry1E, Cry1F, Cry1A/F chimeras, Cry1G, Cry1H, Cry1I, Cry1J, Cry1K, Cry1L, Cry2A, Cry2Ab, Cry2Ae, Cry3, Cry3A variants, Cry3B, Cry4B, Cry6, Cry7, Cry8, Cry9, Cry15, Cry34, Cry35, Cry43A, Cry43B, Cry51Aa1, ET29, ET33, ET34, ET35, ET66, ET70, TIC400, TIC407, TIC417, TIC431, TIC800, TIC807, TIC834, TIC853, TIC900, TIC901, TIC1201, TIC1415, TIC2160, TIC3131, TIC836, TIC860, TIC867, TIC869, TIC1100, VIP3A, VIP3B, VIP3Ab, AXMI-AXMI-, AXMI-88, AXMI-97, AXMI-102, AXMI-112, AXMI-117, AXMI-100, AXMI-115, AXMI-113, and AXMI-005, AXMI134, AXMI-150, AXMI-171, AXMI-184, AXMI-196, AXMI-204, AXMI-207, AXMI-209, AXMI-205, AXMI-218, AXMI-220, AXMI-221z, AXMI-222z, AXMI-223z, AXMI-224z and AXMI-225z, AXMI-238, AXMI-270, AXMI-279, AXMI-345, AXMI-335, AXMI-R1 and variants thereof, IP3 and variants thereof, DIG-3, DIG-5, DIG-10, DIG-657 and a DIG-11 protein.

19. The insect inhibitory composition of claim 14 , defined as comprising a plant cell that expresses said recombinant nucleic acid molecule.

20. A commodity product produced from the plant or part thereof of claim 10 , wherein the commodity product comprises a detectable amount of said recombinant nucleic acid molecule or said pesticidal protein.

21. The commodity product of claim 20 , selected from the group consisting of commodity corn bagged by a grain handler, corn flakes, corn cakes, corn flour, corn meal, corn syrup, corn oil, corn silage, corn starch, corn cereal, and the like, and corresponding soybean, rice, wheat, sorghum, pigeon pea, peanut, fruit, melon, and vegetable commodity products

Assignees

• Monsanto Technology Llc

Inventors

• Bowen David J
• Chay Catherine A
• Howe Arlene R
• Kesanapalli Uma

Classifications

• C07K14/325 Primary

  Bacillus thuringiensis crystal peptides, i.e. delta-endotoxins · CPC title

• G01N2333/43552

  from insects · CPC title

• A01N37/46

  N-acyl derivatives · CPC title

• C12N15/8286

  for insect resistance · CPC title

• G01N33/5308

  for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites · CPC title