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Four strains of bacteria were isolated from the diseased caterpillars of leaf roller, Caloptilia theivora infesting the tea. Analysis of the bacteria based on polyphasic approach such as, growth phase, biochemical tests, whole body protein, crystal protein profiles along with bioassay (i.e. LC50 and LT50 values) established them as different strains of Bacillus (Bacillus sp., CT01, CT02, CT03 and CT04) that were close to Bacillus thuringiensis kurstaki (Btk). Biochemical characteristics of CT01 strain showed positive reaction in lysine decarboxylase, ornithin decarboxylase, urease, Voges-Proskaur and oxidase tests and in utilization of trehalose and glucose. However, CT01 differed from Btk in ONPG, urease, nitrate and oxidase tests; showed difference in utilization tests of arabinose, xylose, cellobiose, mellibiose, saccharose and lactose too. Strain CT02 showed difference with Btk in ONPG, urease and nitrate tests, and in utilization tests of citrate, arabinose, xylose, cellobiose, melibiose and lactose. CT03 strain exhibited difference with Btk in urease and esculin hydrolysis tests, and in utilization of citrate, malonate, arabinose, raffinose and lactose. CT04 strain showed difference with Btk in urease and esculin hydrolysis tests and in utilization of citrate, malonate, cellobiose, glucose and lactose. The doubling time was higher for the strains compard to Btk. When protein composition was analyzed by SDS-PAGE, crystals of CT01 showed one major protein band having the molecular weight 51 kDa while Btk showed the band as 52 kDa. CT02 had two protein bands having molecular weight 37 kDa and 31 kDa. A major protein band of 118 kDa was found in CT03 which was absent in all the three strains and Btk. 38 kDa and 29 kDa protein bands were found in CT04 strain. SDS-PAGE profiles of whole cell protein of CT01, CT03 and CT04 strains as well as that of Btk were similar. However, in CT02 an additional protein band of 34 kDa was found. In vitro studies revealed that among the four strains of Bacillus sp., CT04 was more pathogenic compared to the other three strains and Btk. Low LC50 and LT50 values qualify the strain CT04 more promising for biocontrol.
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