• Deepa I

Entomopathogenic nematodes (EPN) lead a symbiotic association with specific enterobacteria. The nematodes and its specific symbiont are highly pathogenic to a number of insect pests and are being used as biological control agents, worldwide. While working on EPNs, presence of a number of native nematodes belonging to the family Rhabditidae having ability to enter and kill a large number of insect pests in the laboratory within 24 to 48 h of inoculation similar to the EPNs were noticed. Detailed study carried out in our lab indicated that they are associated with specific enterobacteria, which are responsible for the killing action. The bacteria lead a symbiotic association with the nematodes while being highly pathogenic to insect pests. About 65 isolates of such nematodes collected from Kerala, Karnataka, Tamil Nadu, Andhra Pradesh, Bihar, Orissa, New Delhi and Assam are being maintained alive on Galleria mellonella larvae reared on artificial diet. In the present study we have isolated the symbiotic bacteria from surface sterilized infective juveniles of entomopathogenic nematode isolate KO collected from Kovvur, Andhra Pradesh. DNA extraction of these bacteria were carried out and the 16S rDNA fragment was amplified using universal primer. The PCR product was cloned into pGEM-T® vector. The pGEM-T® cloned PCR products were transformed into E.coli (strain DH5α) cells. Transformants were selected by blue/white screening and confirmed by colony PCR, plasmid DNA isolation and its restriction. The selected clones were sent to Delhi university south campus for sequencing. Oligonucleotide sequences obtained from the sequencing reactions were compared with other available sequences in the NCBI GenBank (www.ncbi.nim.nih.gov/Entrez). The nBLAST search showed that the symbiotic bacteria isolated from entomopathogenic nematode isolate KO has 99 % identity with Achromobacter sp. MT-E3 16S ribosomal RNA gene, partial sequence (Accession No. EU727196). The sequences obtained in this study have been assigned in the NCBI GenBank under the accession number HQ200411.

IndexTerms - Entomopathogenic nematodes, 16S ribosomal RNA gene sequence, Polymerase chain reaction (PCR), E.coli, NCBI, Achromobacter sp.

"MOLECULAR IDENTIFICATION OF A NEW SYMBIOTIC BACTERIA ACHROMOBACTER ASSOCIATED WITH ENTOMOPATHOGENIC NEMATODES", International Journal of Emerging Technologies and Innovative Research (www.jetir.org), ISSN:2349-5162, Vol.6, Issue 6, page no.677-680, June 2019, Available :http://www.jetir.org/papers/JETIR1907F92.pdf

"MOLECULAR IDENTIFICATION OF A NEW SYMBIOTIC BACTERIA ACHROMOBACTER ASSOCIATED WITH ENTOMOPATHOGENIC NEMATODES", International Journal of Emerging Technologies and Innovative Research (www.jetir.org | UGC and issn Approved), ISSN:2349-5162, Vol.6, Issue 6, page no. pp677-680, June 2019, Available at : http://www.jetir.org/papers/JETIR1907F92.pdf