Screening of indigenous Bacillus thuringiensis for dipteran active cry gene profiles and potential toxicity against melon fruit fly, Zeugodacus cucurbitae (Coquillett)

Abstract

Abstract Background Melon fruit fly, Zeugodacus cucurbitae Coquillett (Diptera: Tephritidae), is a devastating polyphagous pest attacking large number of fruits and vegetables causing huge economic yield losses across the world. Management of this notorious pest is very challenging as the larvae feed inside the fruit. Hence, the present research study aimed to screen the indigenous Bacillus thuringiensis Berliner (Bacillales: Bacillaceae) strains causing toxicity to larvae and to identify the insecticidal toxicity-related genes present in respective strains. In the present study, 50 indigenous B. thuringiensis (Bt) strains along with one reference strain were screened against second-instar larvae Z. cucurbitae. All the strains were analyzed for presence of 21 dipteran active cry genes. Results Mortality in Z. cucurbitae larvae due to Bt strains ranged from 16 to 92%. PCR results revealed that each strain tested positive for a minimum of three cry genes to maximum of nine cry genes. Among the cry genes, cry1A, cry2A, cry1C, cry19, cry11 and cry70 were detected in high frequency of 100, 88, 84, 74, 58 and 54%, respectively. Bioassay studies revealed that ten out of fifty strains displayed more than 50% mortality. Hence, these ten strains, along with the reference strain, were further tested for mortality for the calculation of the median lethal concentration (LC50). The LC50 values ranged between 38.48 and 105.18 μg/ml. The lowest LC50 found for the strain NBAIR Bt107 was 38.48 μg/ml and was on par with the reference strain (Bti 4Q1) (31.3 μg/ml). Conclusion Indigenous Bt strains displayed a toxicity against the larvae of Z. cucurbitae. The probable dipteran active cry genes responsible for toxicity were interpreted. Thus, the Cry toxins from Bt can play a very important role in the management of Z. cucurbitae.

Keywords

• Zeugodacus cucurbitae
• Bacillus thuringiensis
• Bioassay
• Crystal proteins
• Median lethal concentration