管氏肿腿蜂寄生对黄粉甲硫氧还蛋白基因转录的影响  

Transcriptional Regulation of Tenebrio molitor Thioredoxin Gene by Scleroderma guani Parasitization

作　　者：黄镜梅 刘乃勇[1] 张祖兵[2] 杨斌[1] 朱家颖[1] 

机构地区：[1]西南林业大学云南省森林灾害预警与控制重点实验室,云南昆明650224 [2]云南省热带作物科学研究所,云南景洪666100

出　　处：《西南林业大学学报（自然科学）》2016年第6期112-116,共5页Journal of Southwest Forestry University：Natural Sciences

基　　金：国家自然科学基金项目(31260449)资助;云南省林学一级学科博士点建设项目(林学学位授权学科项目经费)资助;云南省中青年学术与技术带头人后备人才项目(2013HB077)资助;云南省高校森林有害生物科技创新团队项目资助

摘　　要：硫氧还蛋白参与体内必要的抗氧化作用和氧化还原调控过程,在昆虫体内能维持稳定的氧化还原状态,为初步了解硫氧还蛋白在黄粉甲体内的作用以及管氏肿腿蜂寄生对该蛋白基因转录的影响,采用RACE技术,克隆获得黄粉甲硫氧还蛋白基因,并进行多序列比对及荧光定量PCR分析。结果表明:cDNA序列长531 bp,开放阅读框318 bp,其推导的氨基酸序列编码106个氨基酸,3'端非编码区序列为95 bp,5'端非编码区序列为119 bp;预测理论分子量为11.97 kDa,等电点为4.22,含有一个保守的氧化还原活性位点CGPC。黄粉甲硫氧还蛋白基因编码的蛋白序列与其他昆虫的硫氧还蛋白相似性高于60%;硫氧还蛋白基因在黄粉甲各发育阶段中,在蛹期的表达水平最高,在成虫期表达水平最低,在蛹期时的脂肪体中高度表达。被管氏肿腿蜂寄生后,黄粉甲硫氧还蛋白基因转录水平明显受到上调诱导。Thioredoxin proteins maintain the stability of the redox state and are involved in essential antioxida-tion and oxidation reduction process regulation in insects. In order to reveal the functions of the thioredoxin proteins gene and its transcription regulated by Scleroderma guani parasitization, this study cloned the full-length cDNA se-quence of thioredoxin gene from Tenebrio molitor by rapid amplification of cDNA ends strategy. The full-length cDNA sequence of thioredoxin gene was 531 bp, and contained a 318 bp open reading frame encoding 106 amino acid res-idues, with 3′and 5′untranslated regions of 95 bp and 119 bp, respectively. Its predicted molecular weight and isoelectric point were 11. 97 kDa and 4. 22, respectively. Its deduced amino acid sequence contained a highly con-served CGPC active site at the N-terminus. Multiple sequence alignment analysis showed that thioredoxin of T. molitor shared over 60% amino acid identities with those of other insects. qPCR analysis showed that thioredoxin gene was expressed at different developmental stages of T. molitor with the highest expression in the pupa and the lowest in the adult. This gene highly expressed in fat body of pupae. After 6 h parasitization by S. guani, the relative expression of thioredoxin gene was obviously up-regulated.

关 键 词：黄粉甲 硫氧还蛋白 基因克隆 序列分析 管氏肿腿蜂 

分 类 号：S763.4[农业科学—森林保护学]