春尺蠖几丁质酶AcinCht10基因的克隆及在逆境胁迫下的表达.

【Objective】The present paper aimed to explore the molecular mechanism of Apocheima cinerarius response to stress and explore the main associated genes about low temperature and starvation. 【Method】This study cloned and obtained the chitinase gene of A. cinerarius, named AcinCht10（GenBank accession No. : OK504624）, the molecular characteristics and basic physicochemical properties of amino acids of this gene were analyzed, sequence alignment was carried out and phylogenetic tree was constructed. And the expression profile of this gene under stress（low temperature and starvation） was analyzed. 【Result】The predicted molecular formula of AcinCht10 gene was C5017H7689N1349O1490S39, the length of coding protein region was 2967 bp, encoding a total of 988 amino acids, the predicted molecular weight of AcinCht10 was 111. 99 kDa, and the predicted isoelectric point was 6. 25. The hydrophilic coefficient of the protein was-0. 547, and the predicted instability coefficient was 37. 19, indicating that it was stable hydrophilic protein. One signal peptide was found, but no transmembrane structure was found. Two catalytic regions and one chitin binding domain were found. There were 48 serine, 43 threonine and 14 tyrosine phosphoric acid sites, no glycosylation sites were detected. Four phosphoric acid sites were detected. Sequence alignment results showed that the consistency of chitinase gene between AcinCht10 and other Lepidopteran insects was higher than 91%. Phylogenetic results showed that the chitinase genes were grouped into 8 groups（type I-VIII）. AcinCht10 was grouped with Manduca sexta Cht10, Trichoplusia ni Cht10, and Ostrinia furnacalis Cht10, which were also lepidoptera. The results of gene expression analysis showed that AcinCht10 gene expression decreased with the increase of starvation treatment time, and there was no significant difference between adjacent treatments. The expression of AcinCht10 gene was the highest under 25 ℃, while the expression of AcinCht10 gene was at a low level under other temperature stresses with no significant difference. 【Conclusion】AcinCht10 gene was closely related to the response to low temperature and starvation stress. The results are helpful to reveal the molecular function of chitinase gene of A. cinerarius under stress, and lay a foundation for further revealing the molecular mechanism of A. cinerarius response to stress. [ABSTRACT FROM AUTHOR]