Response of CDA5 in Hyphantria cunea to Bt toxin ingestion and Knockdown in transfected Sf9 cells

Abstract

Chitin deacetylases (CDAs) are critical enzymes required for chitin degradation, which play an important role in modifying the physical properties of insect cuticles and peritrophic matrices. A novel chitin deacetylase gene (hccda5) was identified from Hyphantria cunea. The recombinant expressed HcCDA5 protein, containing a chitin deacetylase domain (CDAD), did not show chitin deacetylase activity, but instead showed a strong chitin‐binding activity. Transcriptional analysis using qRT‐PCR indicated that hccda5 was mainly expressed in the midgut of H. cunea larvae. Feeding H. cunea larvae with Bt (Bacillus thuringiensis) Cry1A protein induced its downregulation. The gene of hccda5 was successfully silenced by different dsRNAs in insect cell line (Spodoptera frugiperda line, Sf9), which provided a tool to study the function of an expressed gene in insect cell line and laid a foundation for RNAi of hccda5 in H. cunea larvae.

Identification and Functional Study of Chitin Metabolism and Detoxification-Related Genes in Glyphodes pyloalis Walker (Lepidoptera: Pyralidae) Based on Transcriptome Analysis

“…The study of CHS has been widely reported, and the expression of Locusta migratoria CHS2 is mainly distributed in the midgut [14], which is consistent with the expression of GpCHSB detected in this study (Table 2, Figure 2). The relatively high expression level of GpCDA5 in the midgut is consistent with HcCDA5 in Hyphantria cunea, which is mainly expressed in the midgut of H. cunea larvae [35]. GpCHT2b and GpCHT3b showed a highly evolutionary relationship with CHT2 and CHT3 of other species (Figure 3), respectively, indicating that they have played a role in chitin degradation in G. pyloalis.…”