Transgenic plant-based methods for plant pests using RNAI

• US 9,957,523 B2
• Filed: 07/07/2014
• Issued: 05/01/2018
• Est. Priority Date: 09/16/2005
• Status: Expired due to Fees

• Current Assignee: deVGen NV (China National Chemical Corporation)
• Original Assignee: deVGen NV (China National Chemical Corporation)
• Inventors: Raemaekers, Romaan, Feldmann, Pascale, Nooren, Irene, Plaetinck, Geert, Pecqueur, Frederic, Van Bleu, Els, Bogaert, Thierry
• Primary Examiner(s): Zheng, Li
• Application Number:
  US14/324,351
• Publication Number:
  US 20150128303A1
• Time in Patent Office: 1,394 Days
• Field of Search: 800279
• CPC Class Codes: A01N 57/10 having phosphorus-to-oxygen bonds or phosphorus-to-sulfur bonds A01N57/02 takes precedence
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/8218 Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]
• CPC Class Codes: C12N 15/8281 for bacterial resistance
• CPC Class Codes: C12N 15/8282 for fungal resistance
• CPC Class Codes: C12N 15/8283 for virus resistance
• CPC Class Codes: C12N 15/8285 for nematode resistance
• CPC Class Codes: C12N 15/8286 for insect resistance
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: Y02A 40/146 Genetically Modified [GMO] plants, e.g. transgenic plants

Abstract

The present invention relates to methods for controlling pest infestation using double stranded RNA molecules. The invention provides methods for making transgenic plants that express the double stranded RNA molecules, as well as pesticidal agents and commodity products produced by the inventive plants.

Claims

1. An isolated double stranded RNA molecule comprising annealed complementary strands, wherein at least one of said strands comprises a polyribonucleotide

fully complementary to at least 21 contiguous nucleotides of a target gene represented by SEQ ID NO:

3 or 168

wherein ingestion of said double stranded RNA molecule by a plant insect pest inhibits the growth of said insect pest.

2. A cell transformed with a polynucleotide or set of polynucleotides encoding the double stranded RNA molecule of claim 1.

3. The cell of claim 2, wherein said cell is a plant cell.

4. A plant transformed with a polynucleotide or set of polynucleotides encoding the double stranded RNA molecule of claim 1.

5. A plant seed comprising the double stranded RNA molecule of claim 1.

6. A product produced from the plant of claim 4, wherein said product comprises the double stranded RNA molecule of claim 1.

7. The product of claim 6, wherein said product is selected from a group consisting of food, feed, fiber, paper, meal, protein, starch, flour, silage, coffee, tea, and oil.

8. A plant comprising the double stranded RNA of claim 1, wherein said target gene is derived from a pest species selected from the group consisting of insects, arachnids, crustaceans, fungi, bacteria, viruses, nematodes, flatworms, roundworms, pinworms, hookworms, tapeworms, trypanosomes, schistosomes, botflies, fleas, ticks, mites, and lice.

9. The plant of claim 8, wherein said polyribonucleotide inhibits an insect pest biological activity.

10. The plant of claim 8, wherein said polyribonucleotide inhibits expression of said target gene.

11. The plant of claim 10, wherein said target gene is an insect gene.

12. The plant of claim 8, wherein said plant is cytoplasmic male sterile.

13. A method for controlling insect pest infestation, comprising providing an insect pest with plant material comprising the double stranded RNA molecule of claim 1, wherein said double stranded RNA molecule inhibits expression of a nucleotide sequence which is an ortholog of Drosophila melanogaster gene CG8055.

14. A pesticide comprising a plant expressing a polynucleotide or set of polynucleotides encoding the double stranded RNA molecule of claim 1.

15. A method for controlling insect pest infestation, comprising:

(a) introducing a polynucleotide or set of polynucleotides into a plant; and

(b) providing said plant, or portion thereof, to said insect pest, wherein said polynucleotide or set of polynucleotides encodes the double stranded RNA molecule of claim 1.

16. A method for controlling insect pest infestation, comprising:

a) searching for a target gene in said insect pest orthologous to the gene represented by SEQ ID NO;

3, or a gene encoding the amino acid sequence represented by SEQ ID NO;

4;

b) introducing a polynucleotide or set of polynucleotides into a plant; and

c) providing said plant, or portion thereof, to said insect pest, wherein said polynucleotide or set of polynucleotides encodes the double stranded RNA molecule of claim 1.

17. A method for improving crop yield, comprising:

a) introducing a polynucleotide or set of polynucleotides into a plant; and

b) cultivating said plant to allow polynucleotide expression, wherein said expression inhibits feeding by an insect pest and loss of yield due to insect pest infestation, and wherein said polynucleotide or set of polynucleotides encodes the double stranded RNA molecule of claim 1.

18. The method of claim 17, wherein said double stranded RNA molecule suppresses a target gene in an insect pest that has ingested a portion of said crop plant, wherein said double stranded RNA molecule inhibits expression of a nucleotide sequence which is an ortholog of Drosophila melanogaster gene CG8055.

19. A method for producing a commodity product, comprising:

a) introducing a polynucleotide or set of polynucleotides into a plant cell;

b) growing said plant cell under conditions suitable for generating a plant; and

c) producing a commodity product from said plant or part thereof, wherein said polynucleotide or set of polynucleotides encodes the double stranded RNA molecule of claim 1.

20. A method according to claim 13, wherein said plant is selected from the group consisting of acacia, alfalfa, apple, apricot, artichoke, ash tree, asparagus, avocado, banana, barley, beans, beet, birch, beech, blackberry, blueberry, broccoli, Brussels sprouts, cabbage, canola, cantaloupe, carrot, cassava, cauliflower, cedar, a cereal, celery, chestnut, cherry, Chinese cabbage, citrus, clementine, clover, coffee, cotton, cowpea, cucumber, cypress, eggplant, elm, endive, eucalyptus, fennel, figs, fir, geranium, grape, grapefruit, groundnuts, ground cherry, gum hemlock, hickory, kale, kiwifruit, kohlrabi, larch, lettuce, leek, lemon, lime, locust, pine, maidenhair, maize, mango, maple, melon, millet, mushroom, mustard, nuts, oak, oats, okra, onion, orange, an ornamental plant or flower or tree, papaya, palm, parsley, parsnip, pea, peach, peanut, pear, peat, pepper, persimmon, pigeon pea, pine, pineapple, plantain, plum, pomegranate, potato, pumpkin, radicchio, radish, rapeseed, raspberry, rice, rye, sorghum, sallow, spinach, spruce, squash, strawberry, sugar beet, sugarcane, sunflower, sweet potato, sweet corn, tangerine, tea, tobacco, tomato, trees, triticale, turf grasses, turnips, a vine, walnut, watercress, watermelon, wheat, yams, yew, and zucchini.

21. A method for treating insect infestation of plants comprising making a cell according to claim 3 and regenerating a plant from said cell.

22. A method for treating nematode infestation of plants comprising making a cell according to claim 3 and regenerating a plant from said cell.

23. An isolated polynucleotide or set of polynucleotides encoding the double stranded RNA molecule of claim 1.