Identification, Characterization, and Functional Analysis of Chitin Synthase Genes in Glyphodes pyloalis Walker...

Glyphodes pyloalis Walker (G. pyloalis) causes significant damage to mulberry every year, and we currently lack effective and environmentally friendly ways to control the pest. Chitin synthase (CHS) is a critical regulatory enzyme related to chitin biosynthesis, which plays a vital role in the growth and development of insects. The function of CHS in G. pyloalis, however, has not been studied. In this study, two chitin synthase genes (GpCHSA and GpCHSB) were screened from our previously created transcriptome database. The complete coding sequences of the two genes are 5,955 bp and 5,896 bp, respectively. Expression of GpCHSA and GpCHSB could be detected throughout all developmental stages. Relatively high expression levels of GpCHSA occurred in the head and integument and GpCHSB was most highly expressed in the midgut. Moreover, silencing of GpCHSA and GpCHSB using dsRNA reduced expression of downstream chitin metabolism pathway genes and resulted in abnormal development and wings stretching, but did not affect normal pupating of larvae. Furthermore, the inhibitor of chitin synthesis diflubenzuron (DFB) was used to further validate the RNAi result. DFB treatment significantly improved expression of GpCHSA, except GpCHSB, and their downstream genes, and also effected G. Pyloali molting at 48 h (62% mortality rate) and 72 h (90% mortality rate), respectively. These results show that GpCHSA and GpCHSB play critical roles in the development and wing stretching in G. pyloalis adults, indicating that the genes are attractive potential pest control targets.