昆虫学报2016，Vol.59Issue(9)：938-947,10.DOI:10.16380/j.kcxb.2016.09.003
柞蚕蛹解除滞育过程中海藻糖合成酶基因的表达变化
Changes in the expression of trehalose-6-phosphate synthase gene in Antheraea pernyi (Lepidoptera: Saturniidae) during pupal diapause termination
摘要
Abstract
[Aim] This study aims to clone the trehalose-6-phosphate synthase (TPS) gene in Antheraea pernyi,to analyze its tissue distribution,and to profile the expression patterns of the gene during diapause termination so as to provide data support for clarifying the relationship between carbohydrate metabolism and diapause termination in diapause pupae of A.pernyi.[Methods] The TPS gene was cloned from the fat body of A.pernyi larvae using PCR and 3'RACE technology,and the putative amino acid sequence was analyzed by bioinformatics methods.The expression pattern of the gene in different tissues of A.pernyi larvae was analyzed by RT-PCR.The relative expression levels of the gene in haemolymph and fat body in the process of diapause termination of A.pernyi larvae were measured using real-time PCR technology.[Results] The TPS gene of A.pernyi was cloned and named ApTPS.It has an open reading frame (ORF) of 2 487 bp,encoding 828 amino acid residues.The encoded protein is estimated to be 93.19 kD with an isoelectric point (pI) about 4.61,and has no signal peptide and transmembrane region.Subcellular localization analysis indicated that ApTPS targets to cytoplasm.Homology search showed that ApTPS has two conserved domains:TPS (22-497 amino acids) and TPP (532-772 amino acids).Tissue-specific mRNA expression profiling showed that ApTPS was mainly expressed in the fat body of A.pernyi larvae.In the process of diapause termination,the expression levels of ApTPS in both haemolymph and fat body were up-regulated and significantly higher than those in the control groups (P ＜ 0.05).The elevated expression of the gene in the haemolymph,however,occurred later than that in the fat body.[Conclusion] The results suggest that ApTPS participates in the regulation of carbohydrate metabolism and plays a significant role in diapause process of A.pernyi pupae.There is a close relationship between ApTPS expression and the diapause termination of A.pernyi pupae.关键词
柞蚕/滞育/海藻糖合成酶/基因表达/滞育解除Key words
Antheraea pernyi/diapause/trehalose-6-phosphate synthase/gene expression/diapause termination分类
生物科学引用本文复制引用
黄伶,孙良振,王勇,汝玉涛,Muhammad IRFAN,姜义仁,石生林,杨瑞生,李喜升..柞蚕蛹解除滞育过程中海藻糖合成酶基因的表达变化[J].昆虫学报,2016,59(9):938-947,10.基金项目
现代农业产业技术体系建设专项(CARS-22) （CARS-22）
辽宁省农业领域青年科技创新人才培养计划项目(2014010) （2014010）
辽宁省教育厅一般项目(L2014255,L2015488) （L2014255,L2015488）
