Light chain fibroin and P25 genes of Corcyra cephalonica: molecular cloning, characterization, tissue-specific expression, synchronous developmental and 20-hydroxyecdysone regulation during the last instar larval development

Abstract

The biologically active ecdysteroid hormone, 20-hydroxyecdysone (20E), regulates various processes like molting, metamorphosis and reproduction in insects. However, its role in expression of silk genes is obscure. The silk core in insects is generally constituted of a complex of three proteins namely, H-chain fibroin (H-fibroin), L-chain fibroin (L-fibroin) and P25. In the present study, we report full-length cDNA cloning and characterization of L-fibroin and P25 genes from rice moth, Corcyra cephalonica. Northern analyses demonstrated 1.1 and 1 kb transcripts of L-fibroin and P25 respectively. The tissue expression pattern shows the presence of these transcripts specifically in the salivary gland. These two genes are developmentally regulated at transcriptional level and their maximum expression is observed during the late-last instar larval stage. Semi-quantitative and real-time PCR studies revealed that 20E regulates the expression of these genes in a dose-dependant manner. This study further shows that physiological dose of 20E does not alter the normal expression of these two genes, whereas treatments with higher doses cause a significant decline in the expression. This study clearly suggests the role of 20E in the regulation of L-fibroin and P25 at molecular level.