# US 10,308,954 B2 · Method for the control of nematodes in plants > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,308,954 B2 - Document ID: US10308954B2 - Status: Granted - Kind: B2 - Application number: 14/652,096 - Filed: 2013-12-14 - Published: 2015-12-17 - Granted: 2019-06-04 - Earliest claimed: 2012-12-14 - Assignees: Icar; Department of Biotechnology - Applicants: Icar; Department of Biotechnology - Inventors: Uma Rao; Pradeep Papolu; Nagavara Prasad Gantasala; Divya Kamaraju; Prakash Banakar; Mukesh Kumar - Web page: https://patentgrep.com/patent/US10308954B2 ## CPC - C12N 15/8285: {for nematode resistance} - C07K 14/4354: {from nematodes} - C07K 14/575: Hormones (derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin C07K14/665, e.g. corticotropin C07K14/695) - C12N 15/8218: {Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]} - Y02A 40/146: Genetically Modified [GMO] plants, e.g. transgenic plants - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE ## Abstract 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. ## Claims ### Claim 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. ### Claim 2 (depends on 1) The process as claimed in claim 1, wherein said nematode is Meloidogyne incognita. ### Claim 3 (depends on 1) The process as claimed in claim 1, wherein said plant is from Solanaceous family. ### Claim 4 (depends on 1) The process as claimed in claim 1, wherein said plant is Nicotiana tabaccum or Solanum melongena. ### Claim 5 (depends on 1) 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. ### Claim 6 (depends on 1) 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. ### Claim 7 (depends on 1) The process as claimed in claim 1, wherein said process reduces about 57% in the eggs as compared to the control population. ### Claim 8 (depends on 1) The process as claimed in claim 1, wherein Critical Difference value for the reduction in multiplication index of said nematode is about 1.91. ### Claim 9 (depends on 1) 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. ### Claim 10 (depends on 1) 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. ### Claim 11 (depends on 1) The process as claimed in claim 1, wherein Critical Difference value for the reduction in number of female population is about 1.83. ### Claim 12 (depends on 1) The process as claimed in claim 1, wherein Critical Difference value for the reduction in number of galls is about 1.26. ### Claim 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. ### Claim 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. ### Claim 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.