Methods and compositions for root knot nematode control

Patent metadata

Field	Value
Publication number	US-9624508-B2
Application number	US-201414504330-A
Country	US
Kind code	B2
Filing date	Oct 1, 2014
Priority date	Apr 10, 2008
Publication date	Apr 18, 2017
Grant date	Apr 18, 2017

Abstract

The present invention discloses gene targets, constructs and methods for the genetic control of plant disease caused by nematodes of the genus Meloidogyne (root knot nematodes). The present invention relates to achieving a plant protective effect through the identification of target coding sequences and the use of recombinant DNA technologies for post-transcriptionally repressing or inhibiting expression of the target coding sequences in the cells of plant-parasitic nematodes. The disclosed gene targets show significant conservation at the nucleotide level between orthologs from different Meloidogyne species, facilitating genus-wide targeting by RNA interference.

First claim

What is claimed is:

1. A polynucleotide encoding a double stranded ribonucleotide molecule, the ribonucleotide molecule comprising:
(a) a fragment of at least 23 contiguous nucleotides of SEQ ID NO:40; and
(b) the reverse complement of the fragment of (a), wherein uptake by a plant-parasitic nematode of the ribonucleotide molecule inhibits plant-parasitic nematode infestation.

2. The polynucleotide of claim 1 , defined as operably linked to a heterologous promoter.

3. The polynucleotide of claim 1 , comprising a spacer polynucleotide sequence between the fragment and the reverse complement of the fragment.

4. A transformation vector comprising the polynucleotide of claim 1 .

5. The transformation vector of claim 4 , wherein the polynucleotide is operably linked to a heterologous promoter functional in a plant cell.

6. A double stranded ribonucleotide molecule produced from the expression of a polynucleotide according to claim 1 .

7. The double stranded ribonucleotide molecule of claim 6 , wherein the ribonucleotide molecule inhibits the expression of a target gene in the plant-parasitic nematode.

8. A cell transformed with the polynucleotide of claim 1 .

9. The cell of claim 8 , defined as prokaryotic cell.

10. The cell of claim 8 , defined as a eukaryotic cell.

11. The cell of claim 8 , defined as a plant cell.

12. A plant transformed with the polynucleotide of claim 1 .

13. The plant of claim 12 , further defined as selected from a crop of the group consisting of: corn, wheat, barley, rye, rice, potato, tomato, cucumber, pepper, clover, legume, soybean, pea, alfalfa, sugar cane, sugar beet, tobacco, carrot, cotton, rapeseed (canola), sunflower, safflower, sorghum, strawberry, banana, turf, and an ornamental.

14. A seed of the plant of claim 12 , wherein the seed comprises the polynucleotide.

15. The plant of claim 12 , wherein the polynucleotide is expressed in the plant as a double stranded ribonucleotide molecule.

16. The plant of claim 15 , wherein the uptake by a plant-parasitic nematode of tissue of the plant comprising the double stranded ribonucleotide molecule inhibits plant-parasitic nematode infestation.

17. The plant of claim 16 , wherein the plant-parasitic nematode is a Meloidogyne spp.

18. The plant of claim 16 , wherein the plant-parasitic nematode is Meloidogyne incognita.

19. A commodity product produced from a plant according to claim 12 , wherein the commodity product comprises a detectable amount of the polynucleotide or a ribonucleotide expressed therefrom.

20. A method for controlling a plant-parasitic nematode population comprising providing an agent comprising a double stranded ribonucleotide molecule that functions upon being taken up by the nematode to inhibit a biological function within the nematode, wherein the double stranded ribonucleotide molecule comprises at least 23 contiguous nucleotides of SEQ ID NO:40 and the reverse complement thereof.

21. A method for controlling a plant-parasitic nematode population comprising providing an agent comprising a first polynucleotide sequence comprising at least 23 contiguous nucleotides of SEQ ID NO:40 or complements thereof, wherein the first polynucleotide is hybridized to a second polynucleotide sequence that is complementary to the first polynucleotide sequence.

22. The method of claim 21 , wherein the nematode is Meloidogyne spp.

23. The method of claim 21 , wherein the nematode is Meloidogyne incognita.

24. A method for controlling a plant-parasitic nematode population comprising providing in the host plant of a plant-parasitic nematode a transformed plant cell expressing a polynucleotide molecule according to claim 1 , wherein the polynucleotide is expressed to produce a double stranded ribonucleic acid that functions upon being taken up by the plant-parasitic nematode to inhibit the expression of a target sequence within the nematode and results in decreased growth or reproduction of the nematode or nematode population, relative to growth or reproduction on a host plant lacking the transformed plant cell.

25. The method of claim 24 , wherein the plant-parasitic nematode population exhibits decreased growth following infection of the host plant.

26. The method of claim 24 , wherein the target sequence encodes a protein, the predicted function of which is selected from the group consisting of: egg production and reproduction.

27. The method of claim 24 , wherein said nematode comprises Meloidogyne spp.

28. The method of claim 24 , wherein the nematode is Meloidogyne incognita.

29. The method of claim 24 , wherein the polynucleotide functions upon being taken up by the plant-parasitic nematode to suppress expression of a gene that performs a function essential for egg production.

30. A method for reducing the number of root knot nematode (RKN) feeding sites established in the root tissue of a host plant, comprising providing in the host plant of a Meloidogyne spp. a transformed plant cell expressing a polynucleotide according to claim 1 , wherein the polynucleotide is expressed to produce a double stranded ribonucleic acid that functions upon being taken up by the Meloidogyne spp. to inhibit the expression of a target sequence within said nematode and results in a decrease in the number of feeding sites established, relative to the number of feeding sites established on a host lacking the transformed plant cell.

31. A method of controlling plant nematode pest infestation in a plant comprising providing in the diet of a plant nematode pest a dsRNA comprising the polynucleotide of claim 1 .

32. The method of claim 31 , wherein said diet comprises a plant cell transformed to express said polynucleotide.

33. A method for improving the yield of a crop produced from a crop plant subjected to plant-parasitic nematode infection, said method comprising the steps of
a) introducing a polynucleotide according to claim 1 into said crop plant;
h) cultivating the crop plant to allow expression of said polynucleotide;
wherein expression of the polynucleotide inhibits plant-parasitic nematode infection, growth, reproduction, or loss of yield due to plant-parasitic nematode infection.

34. The method of claim 33 , wherein the crop plant is selected from the group consisting of: corn, wheat, barley, rye, rice, potato, tomato, cucumber, pepper, clover, legume, soybean, pea, alfalfa, sugar cane, sugar beet, tobacco, carrot, cotton, rapeseed (canola), sunflower, safflower, sorghum, strawberry, banana, turf, and an ornamental.

35. The method of claim 33 , wherein expression of the polynucleotide produces an RNA molecule that suppresses at least a first target gene in a plant-parasitic nematode that has contacted a portion of said crop plant, wherein the target gene performs at least one essential function selected from the group consisting of egg production and reproduction.

36. The method of claim 35 , wherein the plant-parasitic nematode is a Tylenchid nematode.

37. The method of claim 36 , wherein the plant-parasitic nematode is a Meloidogyne spp.

38. The method of claim 37 , wherein the plant-parasitic nematode is Meloidogyne incognita.

39. A method for impr

Assignees

• Monsanto Technology Llc

Inventors

• Bradley John D
• Chiapelli Brandi J
• Gao Bingli
• Mccarter James P
• Gasper Michelle L
• Williams Deryck Jeremy

Classifications

• C12N15/8218

  Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS] · CPC title

• A01N57/16

  containing heterocyclic radicals · CPC title

• C12N15/8285 Primary

  for nematode resistance · CPC title

• A01N65/00

  Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof (containing compounds of determined constitution A01N27/00 - A01N59/00) · CPC title

• C12N2310/14

  interfering nucleic acids [NA] · CPC title