Analysis of transcription differences of the heat shock cognate 70 gene in different tissues in response to heat stress in Xestia c-nigrum Linnaeus (Lepidoptera: Noctuidae)
Author of the article:WANG Ling1** YANG Shuai2 ZHU Ming-He1 XU Zhong-Xin1WANG Ling1** YANG Shuai2 ZHU Ming-He1 XU Zh
Author's Workplace：1. College of Agriculture, Northeast Agricultural University, Harbin 150030, China; 2. Virus-Free Seedling Research Institute,Heilongjiang Academy of Agricultural Sciences, Harbin 150086, China
Key Words：Xestia c-nigrum Linnaeus, HSC70, Real-time qPCR, western blotting
Abstract：[Objectives] To investigate the effects of heat on Xe-hsc70 expression in Xestia c-nigrum Linnaeus. [Methods] cDNA and gDNA sequences of the heat shock cognate 70 (70 ku heat shock cognate, HSC70) gene ( hereafter Xe-hsc70) were cloned from fourth instar larvae of X. c-nigrum using the RT-PCR and RACE techniques. The relative expression levels of mRNA and protein in different tissues (malpighian tubules, fat body, midgut, epidermis and salivary glands) under a series temperatures and times were determined by real time quantitative PCR and Western blotting. [Results] The Xe-hsc70 gene contains 8 introns, the longest of which lies at 5′-UTR and contains a HSE-like core sequence (gaatatgCaGAAtgTTCcaGaa). The other introns (with lengths from 86 to 218 bp) are located in the ORF. Xe-HSC70 expression was highest in the fat body and lowest in the salivary glands at 25℃. The relative expression levels of Xe-HSC70 in the midgut, salivary glands and epidermis increased significantly after heat shock, and increased with the duration of heat stress; expression levels first increased and then returned to the control level. There was no significant difference in the mRNA expression levels of Xe-HSC70 in the fat body and malpighian tubules compared with the control. [Conclusion] These findings indicate that different tissues of X. c-nigrum respond differently to heat stress. The high expression of the Xe-hsc70 gene plays an important role in the high temperature tolerance of X. c-nigrum, and provides a foundation for further investigation of thermal stress-avoidance mechanisms of X. c-nigrum at the molecular level