Chilo partellus is an invasive polyphagous pest that has not been effectively managed with chemical pesticides. To select potential dsRNAs for use in an alternate control strategy, it is crucial to identify and evaluate stable reference genes for knockdown expression studies. This study evaluates the expression stability of seven candidate reference genes in C. partellus larvae fed on crude bacterially-expressed dsRNAs and purified dsRNAs at different time intervals, as well as the developmental stages and sexes. The expression stabilities of the reference genes were evaluated with different software programmes, such as BestKeeper, NormFinder, deltaCt, geNorm, and RefFinder. The overall results rank ELF as the most stably expressed reference gene when larvae were fed with crude bacteria-induced dsRNAs and purified dsRNA. However, Tubulin and HSP70 were more stable under different developmental stages and sexes. The expression levels of larvae that were fed crude bacteria-induced dsRNAs of Chitinase and Acetylcholinesterase were normalized with the four most stable reference genes (ELF, HSP70, V-ATPase and Tubulin) and the least stable reference gene (18S and HSP70) based on the geNorm algorithm. The least stable reference gene showed inconsistent knockdown expression, thereby confirming that the validation of a suitable reference gene is crucial to improve assay accuracy for dsRNA-targeted gene selection in C. partellus.

RNAi-based insecticides for crop protection have witnessed rapid improvement over the years. However, their potential to efficiently control maize stem borer (Chilo partellus) pests has remained underexplored. In this study, double-stranded C. partellus chitinase (dsCHI) toxicity was investigated in C. partellus larvae. Furthermore, we developed transgenic maize lines expressing dsRNA targeted against C. partellus chitinase transcripts and performed detached leaf insect feeding bioassays. Our results revealed that C. partellus chitinase transcript expression was significantly downregulated by 57% and 82% in the larvae. Larvae exhibited various phenotypic distortion levels across developmental stages, and 53% mortality occurred in transgenic fed larvae compared to those fed on nontransgenic leaves. In conclusion, we have identified the C. partellus chitinase gene as a potential target for RNAi-mediated control and demonstrated that oral delivery via bacteria and plant-mediated delivery are viable means of achieving C. partellus RNAi-mediated control.

Food production is adversely affected by numerous biotic and abiotic factors that lead to reduction in yield and poor quality of the food products. Use of commercially synthetic pesticides was the most common method for pest control in many agricultural crops during recent decades. These synthetic chemicals have effects on all living organism when they consume such crops treated with pesticides. This research attempted is a green regulation technology as an alternative method to control cabbage moth Plutella xylostella towards the reduction of release of toxic chemical and residue. Virulence studies on different strains (Bb6, Bb11, Bb115, Bb116 and Bb362) of Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales) were evaluated. Various doses at 0 (control) 10(4), 10(5), 10(6), 10(7), 10(8) and 10(9) conidies ml(-1) of five strains were applied topically on the third stage larvae of P. xylostella. Different parameters of larvae were measured in terms of mortality, sporulation rates, the number of pupae that emerged as adult, number of eggs laid between survived adults and the survival rate of larvae were examined at different doses, statistical analysis was performed using Cox-regression. We found that Bb11 strain of Beauveria bassiana (Balsamo) produced highest virulence compared to other strains at 10(9) conidia/ml while, Bb6 strains showed lower virulence effect at 10(9) conidia/ml as compared to control dose. Due to the larvicidal effect of different fungus strains, the percentage weight of female adult decreased significantly as compare to the control.