Method of screening transgenic plants for anthelmintic activity

1. A method of screening a transgenic plant for anthelmintic activity, comprising contacting a transgenic plant, with a plurality of nematodes under conditions effective to determine whether or not said plant has anthelmintic activity, wherein said transgenic plant contains at least one DNA construct, said construct comprising:

a) a nucleic acid encoding a polypeptide catalyzes the conversion of a substrate to a C16, C18, or C20 monounsaturated fatty acid product selected from the group consisting of:

wherein X is hydrogen, CoA, glycerol, a monoglyceride, a diglyceride, ACP, methyl, Na+, phosphatidylcholine, or phosphatidylethanolamine, wherein both R1 and R2 are hydroxyl, one of R1 and R2 is hydroxyl and the other is hydrogen, or one of R1 and R2 is keto and the other is hydrogen, and wherein R3 is C2, C4, or C6 alkyl; and

wherein X is hydrogen, CoA, glycerol, a monoglyceride, a diglyceride, ACP, methyl, Na+, phosphatidyleholine, or phosphatidylethanolamine, and wherein R3 is C2, C4, or C6 alkyl, wherein said nucleic acid has 95% or greater identity to the nucleotide sequence as set forth in SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:130, SEQ ID NO:133 or SEQ ID NO:129; and

(b) a regulatory element operably linked to said nucleic acid encoding said polypeptide, wherein said regulatory element confers expression in a vegetative tissue of said plant.

2. The method of claim 1 , wherein said nematodes are contacted with one or more roots of said transgenic plant.

3. A method of screening a transgenic plant for anthelmintic activity, comprising contacting tissue from a transgenic plant, with a plurality of nematodes under conditions effective to determine whether or not said plant tissue has anthelmintic activity, wherein said transgenic plant contains at least one DNA construct, said construct comprising:

a) a nucleic acid encoding a polypeptide catalyzes the conversion of a substrate to a C16, C18, or C20 monounsaturated fatty acid product selected from the group consisting of:

wherein X is hydrogen, CoA, glycerol, a monoglyceride, a diglyceride, ACP, methyl, Na+, phosphatidylcholine, or phosphatidylethanolamine, wherein both R1 and R2 are hydroxyl, one of R1 and R2 is hydroxyl and the other is hydrogen, or one of R1 and R2 is keto and the other is hydrogen, and wherein R3 is C2, C4, or C6 alkyl; and

wherein X is hydrogen, CoA, glycerol, a monoglyceride, a diglyceride, ACP, methyl, Na+, phosphatidyleholine, or phosphatidylethanolamine, and wherein R3 is C2, C4, or C6 alkyl, wherein said nucleic acid has 95% or greater identity to the nucleotide sequence as set forth in SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:130, SEQ ID NO:133 or SEQ ID NO:129; and

(b) a regulatory element operably linked to said nucleic acid encoding said polypeptide, wherein said regulatory element confers expression in a vegetative tissue of said plant.

4. The method of claim 3 , wherein said tissue is root tissue.

5. The method of claim 1 or 3 , wherein said nucleic acid comprises the nucleotide sequence as set forth in SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:130, SEQ ID NO:133 or SEQ ID NO:129.

6. The method of claim 1 or 3 , wherein said nucleic acid is selected from the group consisting of SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:130, SEQ ID NO:133 and SEQ ID NO: 129.

7. The method of claim 1 or 3 , wherein said plant is selected from the group consisting of tobacco, tomato, soybean, corn, cotton, rice, wheat, banana, carrot, potato, strawberry and turf grass.

8. The method of claim 1 or 3 , wherein said DNA construct further comprises a DNA encoding a PDAT, DAGAT, lipase, FAD2, or FAD3 polypeptide operably linked to at least one regulatory element that confers expression in vegetative tissue of said plant.

9. The method of claim 1 or 3 , wherein said anthelmintic activity comprises an increase in nematode-killing activity as compared to a corresponding plant lacking said nucleic acid encoding said polypeptide.

10. The method of claim 1 or 3 , wherein said anthelmintic activity comprises a reduction in nematode development as compared to a corresponding plant lacking said nucleic acid encoding said polypeptide.

11. The method of claim 1 or 3 , wherein said anthelmintic activity comprises an increase in nematode sterility as compared to a plant lacking said nucleic acid encoding said polypeptide.

12. The method of claim 1 , wherein said anthelmintic activity comprises a reduction in the ability of a nematode to infect said plant as compared to a plant lacking said nucleic acid encoding said polypeptide.

13. The method of claim 3 , wherein said anthelmintic activity comprises a reduction in the ability of a nematode to infect said plant tissue as compared to a plant lacking said nucleic acid encoding said polypeptide.

14. The method of claim 1 , wherein said anthelmintic activity comprises a reduction in the ability of a nematode to reproduce within said plant as compared to a plant lacking said nucleic acid encoding said polypeptide.

15. The method of claim 3 , wherein said anthelmintic activity comprises a reduction in the ability of a nematode to reproduce within said plant tissue as compared to a plant lacking said nucleic acid encoding said polypeptide.

16. The method of claim 1 or 3 , wherein said nematodes are selected from the group consisting of Meloidogyne spp., Heterodera spp., Pratylenchus spp., Paratrichodorus spp., Tylenchorhychys spp., Globodera spp., Rotylenchulus spp., Hoplolaimus spp., Belonolaimus spp., Longidorus spp., Ditylenchus spp., Xiphinema spp., Helicotylenchus spp., Radopholus spp., Hirschmanniella spp., and Trichodorus spp.

17. A method of screening a transgenic plant for anthelmintic activity, comprising contacting a transgenic plant, with a plurality of nematodes under conditions effective to determine whether or not said plant has anthelmintic activity, wherein said transgenic plant contains at least one DNA construct, said construct comprising:

a) a nucleic acid encoding a polypeptide catalyzes the conversion of a substrate to a C16, C18, or C20 monounsaturated fatty acid product selected from the group consisting of:

wherein X is hydrogen, CoA, glycerol, a monoglyceride, a diglyceride, ACP, methyl, Na+, phosphatidylcholine, or phosphatidylethanolamine, wherein both R 1 and R 2 are hydroxyl, one of R1 and R2 is hydroxyl and the other is hydrogen, or one of R1 and R2 is keto and the other is hydrogen, and wherein R3 is C2, C4, or C6 alkyl; and

wherein X is hydrogen, CoA, glycerol, a monoglyceride, a diglyceride, ACP, methyl, Na+, phosphatidylcholine, or phosphatidylethanolamine, and wherein R3 is C2, C4, or C6 alkyl, wherein said nucleic acid is selected from the group consisting of SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:130, SEQ ID NO:133 and SEQ ID NO:129; and

(b) a regulatory element operably linked to said nucleic acid encoding said polypeptide, wherein said regulatory element confers expression in a vegetative tissue of said plant.

18. The method of claim 17 , wherein said nematodes are contacted with one or more roots of said transgenic plant.

19. A method of screening a transgenic plant for anthelmintic activity, comprising contacting tissue from a transgenic plant, with a plurality of nematodes under conditions effective to determine whether or not said plant tissue has anthelmintic activity, wherein said transgenic plant contains at least one DNA construct, said construct comprising:

a) a nucleic acid encoding a polypeptide catalyzes the conversion of a substrate to a C16, C18, or C20 monounsaturated fatty acid product selected from the group consisting of:

wherein X is hydrogen, CoA, glycerol, a monoglyceride, a diglyceride, ACP, methyl, Na+, phosphatidylcholine, or phosphatidylethanolamine, wherein both R1 and R2 are hydroxyl, one of R1 and R2 is hydroxyl and the other is hydrogen, or one of R1 and R2 is keto and the other is hydrogen, and wherein R3 is C2, C4, or C6 alkyl; and

wherein X is hydrogen, CoA, glycerol, a monoglyceride, a diglyceride, ACP, methyl, Na+, phosphatidylcholine, or phosphatidylethanolamine, and wherein R3 is C2, C4, or C6 alkyl, wherein said nucleic acid is selected from the group consisting of SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:130, SEQ ID NO:133 and SEQ ID NO:129; and

(b) a regulatory element operably linked to said nucleic acid encoding said polypeptide, wherein said regulatory element confers expression in a vegetative tissue of said plant.

20. The method of claim 19 , wherein said tissue is root tissue.

21. The method of claim 17 or 19 , wherein said nematodes are selected from the group consisting of Meloidogyne spp., Heterodera spp., Pratylenchus spp., Paratrichodorus spp., Tylenchorhychys spp., Globodera spp., Rotylenchulus spp., Hoplolaimus spp., Belonolaimus spp., Longidorus spp., Ditylenchus spp., Xiphinema spp., Helicotylenchus spp., Radopholus spp., Hirschmanniella spp., and Trichodorus spp.