BackgroundThe cotton bollworm, Helicoverpa armigera is one of the most important crop pests worldwide. It has developed high levels of resistance to synthetic insecticides, and hence, Bacillus thuringiensis (Bt) formulations are used as a safer pesticide and the Bt genes have been deployed in transgenic crops for controlling this pest. There is an apprehension that H. armigera might develop resistance to transgenic crops in future. Therefore, we studied the role of gut microbes by eliminating them with antibiotics in H. armigera larvae on the toxicity of Bt toxins against this pest.ResultsCommercial formulation of Bt (Biolep®) and the pure Cry1Ab and Cry1Ac toxin proteins were evaluated at ED50, LC50, and LC90 dosages against the H. armigera larvae with and without antibiotics (which removed the gut microbes). Lowest H. armigera larval mortality due to Bt formulation and the Bt toxins Cry1Ab and Cry1Ac was recorded in insects reared on diets with 250 and 500 μg ml−1 diet of each of the four antibiotics (gentamicin, penicillin, rifampicin, and streptomycin), while the highest larval mortality was recorded in insects reared on diets without the antibiotics. Mortality of H. armigera larvae fed on diets with Bt formulation and the δ-endotoxins Cry1Ab and Cry1Ac was inversely proportional to the concentration of antibiotics in the artificial diet. Nearly 30% reduction in larval mortality was observed in H. armigera larvae from F1 to F3 generation when the larvae were reared on diets without antibiotics (with gut microbes) and fed on 0.15% Bt or 12 μg Cry1Ab or Cry1Ac ml−1 diet, indicating development of resistance to Bt in the presence of gut microflora. However, there were no differences in larval mortality due to Bt, Cry1Ab or Cry1Ac across generations in insects when they were reared on diets with 250 μg of each antibiotic ml−1 diet (without gut microflora).ConclusionsThe results suggested that antibiotics which eliminated gut microflora influenced the toxicity of Bt towards H. armigera, and any variation in diversity and abundance of gut microflora will have a major bearing on development of resistance to Bt toxins applied as foliar sprays or deployed in transgenic crops for pest management.
Transgenic crops expressing toxin proteins from Bacillus thuringiensis (Bt) have been deployed on a large scale for management of Helicoverpa armigera. Resistance to Bt toxins has been documented in several papers, and therefore, we examined the role of midgut microflora of H. armigera in its susceptibility to Bt toxins. The susceptibility of H. armigera to Bt toxin Cry1Ac was assessed using Log-dose-Probit analysis, and the microbial communities were identified by 16S rRNA sequencing. The H. armigera populations from nine locations harbored diverse microbial communities, and had some unique bacteria, suggesting a wide geographical variation in microbial community in the midgut of the pod borer larvae. Phylotypes belonging to 32 genera were identified in the H. armigera midgut in field populations from nine locations. Bacteria belonging to Enterobacteriaceae (Order Bacillales) were present in all the populations, and these may be the common members of the H. armigera larval midgut microflora. Presence and/or absence of certain species were linked to H. armigera susceptibility to Bt toxins, but there were no clear trends across locations. Variation in susceptibility of F1 neonates of H. armigera from different locations to the Bt toxin Cry1Ac was found to be 3.4-fold. These findings support the idea that insect migut microflora may influence the biological activity of Bt toxins.
a b s t r a c tStudies on the influence of genotypic resistance on biological activity of a commercial formulation of Bacillus thuringiensis (Bt) and pure Bt toxin Cry1Ac were carried out to develop appropriate strategies for pod borer, Helicoverpa armigera management in chickpea, sorghum, pigeonpea and cotton. The interaction effects of host plant resistance and biological activity of commercial Bt/Cry1Ac were studied by incorporating the lyophilized tissues of chickpea leaves, milk stage sorghum grain, pigeonpea pods and cotton squares into the artificial diet with and without Bt formulation or Cry1Ac. The H. armigera larval weights were significantly lower in insects reared on diets with square powder of the insect -resistant Bt-cotton RCH 2 þ Bt/Cry1Ac and pod powder of insect -resistant pigeonpea genotype, ICPL 332WR þ Bt/ Cry1Ac as compared to the larvae reared on diets with leaf powder of H. armigera susceptible chickpea genotype, ICCC 37 and the standard artificial diet. Pupation and adult emergence were significantly lower in insects reared on diets with tissues of pod borer-resistant genotypes þ Bt/Cry1Ac as compared to insects reared on diets with tissues of the insect susceptible genotypes þ Bt/Cry1Ac. Insects reared on diets containing insect-resistant and -susceptible genotypes of sorghum, pigeonpea and cotton and pod borer-resistant genotype of chickpea (ICC 506EB) þ Bt/Cry1Ac did not lay any eggs. However, eggs were laid by the insects reared on diets containing pod borer-susceptible genotype of chickpea, ICCC 37 and on the standard artificial diet þ Bt/Cry1Ac. The insects reared on diets with sorghum genotype, ICSV 745, and Bt-cotton, RCH 2 without Bt/Cry1Ac also did not lay eggs. The results suggested that Bt/Cry1Ac is more effective for management of H. armigera when deployed in combination with insect-resistant genotypes of cotton, chickpea, pigeonpea and sorghum.
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