Knockdown of β-N-acetylglucosaminidase gene disrupts molting process in Heortia vitessoides Moore.

Abstract

β-N-acetylglucosaminidase (NAG) is a key enzyme in insect chitin metabolism and plays an important role in many physiological activities of insects. The HvNAG1 gene was identified from the Heortia vitessoides Moore (Lepidoptera: Crambidae) cDNA library and its expression patterns were determined using quantitative real-time polymerase chain reaction. The results indicated that HvNAG1 mRNA levels were high in the midgut and before molting, and 20E could induce its expression. Subsequently, the HvNAG1 gene was knocked down via RNA interference to identify its functions. We found that 3 μg of dsNAG1 resulted in optimal interference at 48 and 72 hr after injection, causing a decrease in NAG1 protein content, which resulted in abnormal or lethal phenotypes, and a sharp decrease in the survival rate. These results indicate that HvNAG1 plays a key role in the molting process of H. vitessoides. However, the silencing of HvNAG1 had no significant effect on the chitin metabolism-related genes tested in this study. Our present study provides a reference for further research on the utility of key genes involved in the chitin metabolic pathway in the insect molting process.