Exploration of insecticidal potential of Cry protein purified from Bacillus thuringiensis VIID1

Insect pests are a threat to agriculture as they cause a loss of 15-22% to economically important crops every year. Bacillus thuringiensis produces parasporal crystal inclusions that have insecticidal 'Cry' proteins which are toxic to insect larvae of the order Lepidoptera, Coleoptera and Diptera, etc. In the present study, 40 different soil samples from Amritsar and its surrounding areas were selected for isolation of B. thuringiensis. The rod shaped, gram-positive bacterial isolates were further analyzed for characteristic crystal formation using phase contrast and scanning electron microscopy. 6 Bacillus samples containing cry genes were identified using the universal primers for cry genes, of which one isolate exhibited a protein band of similar to 95 kDa. This protein was purified using a Sephadex G-75 column. The insecticidal assays conducted with purified Cry protein on insect larvae of lepidopteran and dipteran orders viz. Spodoptera litura, Galleria malonella, Bactrocera cucurbitae and Culex pipens revealed considerable detrimental effects. A significant increase in larval mortality was observed for the larvae of all insects in a concentration dependent manner when treated with Cry protein purified from B. thuringenisis VIID1. The purified Cry protein did not have any significant effect on honey bee larvae. (C) 2021 Elsevier B.V. All rights reserved.

Automated summary

Bacillus thuringiensis produces insecticidal proteins that are toxic to insect larvae of economically important crops. In this study, B. thuringiensis was isolated from soil samples in Amritsar, and a protein band of about 95 kDa was identified in one isolate. The purified Cry protein showed significant detrimental effects on lepidopteran and dipteran insect larvae, while having no significant effect on honey bee larvae.