CHARACTERIZATION OF cry GENES IN NATIVE Bacillus thuringiensis ISOLATES

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Date
2000
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University of Agricultural Sciences, Dharwad
Abstract
"The present sUicly was concluded lo isolalc Baciffus thuringiensis from soils o f Westeni Ghat region in Uttara Kannada district o f Karnataka, India, to assess their insecticidal activity, molecular diversity and to identify specific cry genes in them. Out o f the initial 41 isolates, only ten isolates had endospores and crystals. Three isolates i.e., D l , D3 and D 2 I were toxic to diamond back moth {l^lu(ella xylos lella) larvae. Among these isolates D l caused the highest mortality (9 0% ) followed by D3 (8 0% ) and D21 (7 0%) . The isolates varied in sensitivity to antibiotics. All the three isolates, reference strain Bt -4 2 and reference isolate P I were sensitive to tetracycline, chloramphenicol and kanamycin. D l , D3 and D21 were resistant to ampicillin (1 5 0 ppm) and spectinomycin. In addition, D3 was also resistant to nalidixic acid (5 0 ppm). SDS-PAGF'. protein profile indicated the presence o f 130 kD a and 65 kD a bands in D l , D3, Bt -42 and P I . In D21, a 26 kD a protein was present. The genetic distance (% ) based on protein profile indicated the minimum genetic distance o f 16 .6 7% between PI and D l and the maximum genetic distance o f 8 0 .0 0% uciwccn D i and D2 1 . All the isolates had only one plasmid o f about 15 kb. The presence o f c ry genes in the isolates was detected based on the nature o f the PGR amplification products. Wi th c r y specific primers it was found that gene c ry !('. was present in all the three isolates D l , D3 and D21. c ry IA(c) gene was present in PI and D I and the nematode activc c ry gene was detected in P i , D l and D3."
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No. of Reference 56
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