Cloning, Expression and Ecdysone Regulation of Chitinase MsCHT7 from the Oriental Armyworm, Mythimna separata Walker

doi:10.16409/j.cnki.2095-039x.2018.03.004

Abstract

Abstract: Chitinase is a key enzyme in chitin hydrolysis and plays an important role in insect growth and development. In this study, a novel chitinase gene cDNA sequence MsCHT7 (GenBank accession No. MG551526) was obtained from Mythimna separata by high-throughput transcriptome sequencing technology. The full-length cDNA is 3360 bp and encodes an amino acid sequence with 989 residues. The amino acid sequence includes a highly conserved sequence of family 18 glycoside hydrolases, two Glyco_18 catalytic domains and a chitin binding domain ChtBD2. qRT-PCR method was used to reveal stage-and tissue-dependent expression patterns of MsCHT7. The results showed that MsCHT7 expressed specifically at different developmental stages and in different tissues. The expression of MsCHT7 was the highest at prepupal stage and in integuments and higher in last-day pupa and adult stage. The expression levels were higher on the last day than on the first day of each instars. After treatment with 20E (20-hydroxyecdysone), the expression levels of MsCHT7 at different time points showed significant differences. The expression levels of MsCHT7 increased with time and reached the peak at 24 h, then significantly decreased. The MsCHT7 expression varied with four different 20E concentrations. When the concentration was 10 μg/μL, the expression level was the highest. Treatment with different 20E concentrations advanced molting of fifth instar larvae. Because expression of MsCHT7 was regulated by ecdysone, MsCHT7 might play a role in M. separata molting.

Key words: Mythimna separata, chitinase, MsCHT7, expression, ecdysone, regulation

CLC Number:

S435.13

YANG Wenqing, FAN Dong. Cloning, Expression and Ecdysone Regulation of Chitinase MsCHT7 from the Oriental Armyworm, Mythimna separata Walker[J]. journal1, 2018, 34(3): 354-363.

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