Analysis Of Chitin Degradation-related Genes Of Lymantria Dispar Based On High-throughput Sequencing

Lymantria dispar is an important agricultural pests and widely distributed in the world, the chitin metabolism is essential in process of L.dispar growth and development.However, the study of L.dispar genetic information has been relatively scarce.The high-throughput sequencing technology was used to sequence the L.dispar transcriptome and find out genes about enzymes associated with the chitin degradation, then we study the expression and function of they genes.The main work is as follows:(1) This study we anatomy L.dispar and get their skin and intestinal tissue, then extracted its RNA. A total of 36,882 unigenes of this species were obtained by sequencing the transcriptome, then these unigenes by BLAST alignments SWISS-PROT, TREMBL, CDD, PFAM, NR and KOG databases annotated.(2) This study we find seven chitin degradation-related genes from L.dispar transcriptome database, respectively chitinase LdChtⅢ, chitin deacetylases LdCda1, LdCda2, LdCda3 and LdCda.4, β-N-acetylglueosaminidaseLdNag2 and LdNag3, then we design specific primers, then we study these genes relative expression at different developmental stages of L.dispar by use real-time PCR With laboratory has obtained β-actin sequences as reference genes.(3) We using RNAi method further study the functions of these genes, the sixth-instar larva,prepupa and pupa as study.The result shows:different genes have different relativ experssion, chitinase and β -N-acetylglueosaminidase have highly relative expression in the sixth instar larvae and pupae, we believe that these two enzyme degradation chitin in larva period so that L.dispar could develop to the pupal. chitin deacetylase gene showing irregular expression,which maybe chitin deacetylase involved in various periods of L.dispar chitin degradation activity.The relative expression of various genes is a dynamic process,the target gene in each of larvae have different expression.This study we research L.dispar transcriptome and the chitin degradation on molecular biology level. Our study provides a solid foundation for future L.dispar research, have a very important academic significance.