草地贪夜蛾热激蛋白Hsc70基因克隆、表达谱及对不同环境胁迫的响应 Click here to download the full text Citation：姚梦霜,孟建玉,杨昌利,张长禹.草地贪夜蛾热激蛋白Hsc70基因克隆、表达谱及对不同环境胁迫的响应.Journal of Plant Protection,2020,47(4):797-806 Hits: Download times: Author Name Affiliation E-mail YAO Mengshuang  Laboratory for Agricultural Pest Management of the Mountainous Region, Institute of Entomology, Guizhou University, Guiyang 550025, Guizhou Province, China    MENG Jianyu  Guizhou Research Institute of Tobacco Sciences, Guiyang 550081, Guizhou Province, China    YANG Changli  Laboratory for Agricultural Pest Management of the Mountainous Region, Institute of Entomology, Guizhou University, Guiyang 550025, Guizhou Province, China    ZHANG Changyu  Laboratory for Agricultural Pest Management of the Mountainous Region, Institute of Entomology, Guizhou University, Guiyang 550025, Guizhou Province, China  zcy1121@aliyun.com  中文摘要:为探讨草地贪夜蛾Spodoptera frugiperda Hsc70基因对A区段紫外辐射（ultraviolet-A radiation，UV-A）及高、低温胁迫的响应，采用逆转录聚合酶链反应（reverse transcription-polymerase chain reaction，RT-PCR）技术克隆Hsc70基因并对其进行序列分析，利用实时荧光定量PCR（real-time quantitative PCR，RT-qPCR）技术分析不同龄期、不同组织及UV-A和高、低温胁迫下的表达量。结果显示，草地贪夜蛾Hsc70基因（GenBank登录号：MT130717）开放阅读框为1 980 bp，编码659个氨基酸，蛋白质相对分子量为73.12 kD，等电点为5.23；其氨基酸末端序列为KDEL，属于内质网型热激蛋白。草地贪夜蛾Hsc70基因与鳞翅目昆虫Hsc70同源性高。草地贪夜蛾Hsc70表达量在2龄幼虫发育阶段和复眼中表达量最高。UV-A、高、低温胁迫对草地贪夜蛾Hsc70基因表达有明显的诱导作用，随着处理时间的延长，Hsc70基因表达量呈先增加后降低的趋势；UV-A胁迫下，草地贪夜蛾雌、雄成虫Hsc70基因表达量分别在120 min和60 min时达到峰值；36℃胁迫下，草地贪夜蛾雌、雄成虫Hsc70基因表达量分别在120 min和90 min时达到峰值；4℃胁迫下，草地贪夜蛾雌、雄成虫Hsc70基因表达量均在30 min时达到峰值。表明草地贪夜蛾Hsc70基因在草地贪夜蛾适应环境胁迫中发挥着作用。 中文关键词:草地贪夜蛾  Hsc70  基因  克隆  表达分析   cDNA cloning and expression profiling of the heat shock prote in Hsc70 gene and its response to different environmental stresses in fall armyworm Spodoptera frugiperda Author Name Affiliation E-mail YAO Mengshuang  Laboratory for Agricultural Pest Management of the Mountainous Region, Institute of Entomology, Guizhou University, Guiyang 550025, Guizhou Province, China    MENG Jianyu  Guizhou Research Institute of Tobacco Sciences, Guiyang 550081, Guizhou Province, China    YANG Changli  Laboratory for Agricultural Pest Management of the Mountainous Region, Institute of Entomology, Guizhou University, Guiyang 550025, Guizhou Province, China    ZHANG Changyu  Laboratory for Agricultural Pest Management of the Mountainous Region, Institute of Entomology, Guizhou University, Guiyang 550025, Guizhou Province, China  zcy1121@aliyun.com  Abstract:To investigate the molecular mechanism of fall armyworm Spodoptera frugiperda in response to UV-A radiation, temperature stresses of high and low, the reverse transcription-polymerase chain reaction (RT-PCR）were used to the cDNA of Hsc70 gene from S. frugiperda, and the cDNA sequence were analyzed. To explore the expression differences of Hsc70 gene from S. frugiperda in different developmental stages, in different tissues and under UV-A, high and low temperature stresses, the expression level of Hsc70 gene were investigated by real-time quantitative PCR (RT-qPCR). The results showed the open reading frame of the Hsc70 gene from S. frugiperda (GenBank accession no.: MT13071) is 1 980 bp, and 659 amino acids of encoding. The relative molecular of its putative protein is 73.12 kD with an isoelectric point (pI) of 5.23. The amino acid terminal sequence is KDEL, indicating that the protein is an endoplasmic heat shock protein. It was found that Hsc70 gene from S. frugiperda shared a high homology with Hsc70 genes from other species of Lepidoptera. Hsc70 gene had the highest expression levels in the 2nd instar larvae and compound eyes of S. frugiperda. UV-A, temperature stresses of high and low induced obviously the Hsc70 gene expression with increasing treatment time. Under UV-A, the expression levels of Hsc70 gene in female and male adults reached the maximum at 120 min and 60 min, respectively. Under 36℃, the expression levels of Hsc70 gene in female adults and male adults reached the peak at 120 min and 90 min, respectively. Under 4℃, the expression levels of Hsc70 gene in both female and male adults were the highest at 30 min. The differential expression of Hsc70 gene under UV-A radiation, temperature stress of high and low indicates that it plays a pertinent role in the molecular mechanism of S. frugiperda in response to environmental stress. keywords:Spodoptera frugiperda  Hsc70  gene  cloning  expression analysis View Full Text  View/Add Comment  Download reader

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