Molecular Characterization of Steinernema masoodi, S. seemae and Other Indian Isolates of Steinernema spp.

Molecular characterization, studies on Steinernema masoodi, S. seemae, S. carpocapsae (Jammu strain) S. carpocapsae (Lucknow strain) have been carried out through analysis of 18S ITS ribosomal RNA genes. The RFLP analysis was carried out, each PCR product were digested with one of the series of 17 restriction enzymes. The digested DNAs loaded on 1.5% agarose gel, separated by electrophoresis at 130 V for 3 h, stained with ethidium bromide, visualised and photographed under UV light. For S. masoodi and S. seemae DNA sequence didn't show any homology with any DNA sequence of other EPN data base and 12% agarose gel exhibiting ITS band amplicule of both the species different from S. abbasi, S. tami and S. carpocapsae and other species in a public domain and it confirms that both are new species at molecular level The dendogram showing phylogenic position of Lucknow strain of S. carpocapsae has generated by alignment of ribosomal gene sequences to other strains of Steinernema indicated that Jammu strain is close to S. carpocapsaeAccn No.AY 171262, DG302092, AY171283, AY17032 while Lucknow strain comes close to strain A11-AY23 while it is different from the closely linked group S. backanense Acc. No. AY487918 and S. sasonense Acc. No. AY487919.Pair wise matrix of ribosomal gene sequence of the different isolates showing the phylogenetic position of strain which indicate that both the strains can be separated by divergence position. The DNA sequences of ~350 bases thus obtained were compared with the sequences present in the public domain (Gene Bank) for homology studies by BLASTN program. A dendrogram and distance matrix was prepared by using the Clustal W program of DNASTAR software (Lasergene, USA) which shows the phylogenetic position, in case of S. carpocapsae.