Method for the control of nematodes in plants
The present invention relates to the field of double-stranded RNA (ds RNA) mediated gene silencing of root knot nematodes. The invention particularly provides an effective method for reducing the number of reproducing population and the number of progenies per individual of the root knot nematodes. The present invention also relates to host delivered dsRNA for controlling infection of root knot nematodes.
1. A process for controlling the multiplication index of a root knot nematode by reducing the number of reproducing population and number of progeny per individual, said process essentially comprises steps of transforming said host plant cell with a nucleic acid, which express to produce a transcript that forms a double stranded RNA having a SEQ ID No. 1 and the complement thereof and consequently facilitates the inhibition of the expression of mRNA of a target gene or a segment thereof present in said nematode species, wherein said target gene is FMRFamide-like peptides 14 gene or flp-14 gene of said nematode and wherein said process causes reduction of about 78% of the multiplication index of said nematode in the transformed host plant cell as compared to the control population of host cell.
2. The process as claimed in claim 1 , wherein said nematode is Meloidogyne incognita.
3. The process as claimed in claim 1 , wherein said plant is from Solanaceous family.
4. The process as claimed in claim 1 , wherein said plant is Nicotiana tabaccum or Solanum melongena.
5. The process as claimed in claim 1 , wherein said process reduces about 41% of the female population laying eggs as compared to the control population.
6. The process as claimed in claim 1 , wherein said process reduces about 48% in the eggs per egg mass as compared to the control population.
7. The process as claimed in claim 1 , wherein said process reduces about 57% in the eggs as compared to the control population.
8. The process as claimed in claim 1 , wherein Critical Difference value for the reduction in multiplication index of said nematode is about 1.91.
9. The process as claimed in claim 1 , wherein Critical Difference value for the reduction in number of egg per egg mass of said nematode is about 1.27.
10. The process as claimed in claim 1 , wherein Critical Difference value for the reduction in number of egg mass per plant of said nematode is about 1.65.
11. The process as claimed in claim 1 , wherein Critical Difference value for the reduction in number of female population is about 1.83.
12. The process as claimed in claim 1 , wherein Critical Difference value for the reduction in number of galls is about 1.26.
13. A method of transferring a RNAi vector for gene silencing of flp-14 gene of root knot nematodes in host plants for control of root-knot nematodes, said method comprising steps of:
(a) preparing the RNAi vector construct comprising SEQ ID No. 1 and the complement thereof;
(b) transferring the vector of step (a) into an Agrobacterium strain;
(c) infecting the host explants with Agrobacterium of step (b); and
(d) obtaining the transformed host plants.
14. A method of expressing dsRNA transcript for RNAi or gene-silencing of flp-14 gene for controlling root-knot nematodes infection, said method comprising steps of:
(a) preparing an expression vector comprising SEQ ID No.1 and the complement thereof;
(b) transferring the vector of step (a) in a host plant;
(c) obtaining the expression of in vitro and in vivo transcript of dsRNA.
15. Transgenic plants expressing double stranded RNA having SEQ ID No.1 and the complement thereof for controlling infection of root-knot nematodes by RNAi or gene silencing of flp-14 gene of root-knot nematodes.