水稻二化螟小分子量热激蛋白21.3基因的克隆和表达分析  

Molecular cloning and expression analysis of a small heat shock protein gene 21.3 from Chilo suppressalis

作　　者：宋杰 于同英 崔亚东[2] 陆明星[1] 杜予州[1] SONG Jie;YU Tong-Ying;CUI Ya-Dong;LU Ming-Xing;DU Yu-Zhou(College ofHorticulture and Plant Protection ＆ Institute of Applied Entomology,Yangzhou University,Yangzhou 225009,Jiangsu Province,China;Biology and Food Engineering School,Fuyang Normal University,Fuyang 236032,Anhui Province,China)

机构地区：[1]扬州大学园艺与植物保护学院,扬州大学应用昆虫研究所,江苏扬州225009 [2]阜阳师范学院生命科学学院,安徽阜阳236032

出　　处：《环境昆虫学报》2018年第3期505-513,共9页Journal of Environmental Entomology

基　　金：国家自然科学基金项目(31401733,31371937)

摘　　要：小分子量热激蛋白(sHSPs,small heat shock proteins)在昆虫中分布广泛,能影响昆虫的生长繁殖、参与多种生理过程以及增强昆虫的温度耐受性。水稻二化螟Chilo suppressalis(Walker)是水稻上的主要害虫之一,目前在我国局部稻区危害严重。为了深入了解sHSPs在水稻二化螟生长发育及温度耐受性中的功能,利用RT-PCR和RACE技术克隆出一个二化螟小分子量热激蛋白Cs HSP21.3基因。Cshsp21.3的cDNA序列全长为844 bp,其中开放阅读框(ORF)为555 bp,编码184个氨基酸,理论分子量为21.3 kDa,等电点为5.97;基因组结构分析发现,Cshsp21.3缺失内含子。实时定量PCR的实验结果表明:在脂肪体中Cshsp21.3的表达高于其它不同组织器官;在二化螟的不同发育阶段,3龄幼虫Cshsp21.3的表达水平最高,而且雌雄蛹的表达差异显著;高低温不能显著诱导Cshsp21.3的表达,说明该基因对温度胁迫不敏感。总之,Cshsp21.3与二化螟的温度耐受性之间没有密切的联系,但在二化螟的生长发育方面起着重要的作用。Small heat shock proteins（sHSPs） are widely distributed in insects,which can affect the development,various physiologies and thermotolerance of insect. The Chilo suppressalis,one of the important rice pests,induce the dangerous loss of rice in part of rice cultivation regions now. Therefore,in order to further understand the function of s HSPs in the physiology,development and thermotolerance of Chilo suppressalis,a small heat shock protein gene,Cshsp21. 3,was identified using RT-PCR and RACE. The results exhibited Cshsp21. 3 possessed a complete cDNA sequence of 844 bp, which contained 555 bp open reading frame（ORF）,and encoded 184 amino acids,with 21. 3 kDa theoretical molecular weight and 5. 97 the isoelectric point. Genomic structure analyses demonstrated Cshsp21. 3 lacked introns. Quantitative real-time PCR revealed that the expression of Cshsp21. 3 in fat body was highest,comparing to other tissues or organs. In the different developmental stages,the expression of third instar larvae was the highest,and there were significant differences between male and female pupae. Under different temperatures,the mRNA expression levels of Cshsp21. 3 weren＇t induced by high or low temperatures,which indicated it was not sensitive to temperature stress. In conclusion,the study indicated Cshsp21. 3 possess low relation to the thermotolerance of C. suppressalis,but it played an important role in the physiology and development of C. suppressalis.

关 键 词：sHSPs 二化螟 组织器官 发育阶段 温度 表达 

分 类 号：Q963[生物学—昆虫学] S433.4[农业科学—农业昆虫与害虫防治]