Virulence assay and role of Bacillus thuringiensis TS110 as biocontrol agent against the larval stages of rice leaffolder Cnaphalocrocis medinalis .

The bacterial isolate Bacillus thuringiensis TS110 was isolated from the rice field soil of Burdwan district, West Bengal, India. Bioassay test of the bacteria TS110 against 3rd, 4th and 5th instar larvae of Cnaphalocrocis medinalis was carried out. Cut leaf assay, potted plant assay and field assay were done. During filed assay, it has been observed that the LC50 (×107 ) values of TS110 against 3rd, 4th and 5th instar larvae of C. medinalis were 3.77, 5.29, 4.83 and 4.93, 4.42, 4.72 in dry and wet season, respectively. The morphological, biochemical and phylogenetic analysis of the isolate TS110 were done. TS110 was positive for catalase, nitrate reductase, methyl red, voges-proskauer, oxidase, urease, indole, citrate utilization, arginine dihydrolase test, starch, lipid, gelatin, casein, and lecithin hydrolysis test. TS110 showed fermentation test positive for glucose, fructose, mannose, arabinose and trehalose in nutrient broth medium. The antibiotic sensitivity test showed that the bacterial isolate was sensitive to kanamycin (30 µg/disc), nalidixic acid (30 µg/disc), rifampicin (5 μg/disc), doxycycline (30 µg/disc), gatifloxacin (10 µg/disc), vancomycin (30 µg/disc), gentamycin (10 µg/disc), ampicillin (10 µg/disc), ofloxacin (5 μg/disc), levofloxacin (5 μg/disc), streptomycin (10 µg/disc), gentamycin (10 µg/disc). The phylogenetic analysis revealed that TS110 was closely related to different species of B. thuringiensis submitted to the GenBank. On the basis of morpho-physiological and molecular characterization, the bacterial isolate was identified as B. thuringiensis.