温度胁迫下棉花粉蚧热激蛋白基因的表达分析  

Expression analysis of heat shock protein genes in Phenacoccus solenopsis (Hemiptera: Pseudococcidae) under temperature stress

作　　者：陈芳[1] 陆永跃[1] 

机构地区：[1]华南农业大学昆虫学系,广州510642

出　　处：《昆虫学报》2014年第11期1253-1264,共12页Acta Entomologica Sinica

基　　金：广东省自然科学基金项目(S2013010013526);广东省研究生教育创新计划项目(2013JDXM14)

摘　　要：【目的】为了研究热激蛋白Hsp70,Hsp70-4和Hsp90在棉花粉蚧Phenacoccus solenopsis抵抗逆温中的作用。【方法】在测序棉花粉蚧转录组的基础上,分析了该虫热激蛋白Hsp70基因家族的2个序列[Pshsp70（Gen Bank登录号为KJ909505）和Pshsp70-4（Gen Bank登录号为KJ909506）]和Hsp90基因家族的1个序列,[Pshsp90（Gen Bank登录号为KJ909507）],采用实时荧光定量PCR（RT-q PCR）检测了在不同温度（18和32℃恒温,37,39,41,43和45℃热激1 h后26℃恢复1 h）下棉花粉蚧不同发育阶段（2龄若虫、3龄若虫、雌成虫）3种热激蛋白基因的表达量。【结果】Pshsp70 c DNA序列包含1 923 bp的开放阅读框,编码641个氨基酸,理论分子量和等电点分别为70.9k Da和5.65;Pshsp70-4 c DNA序列包含1 962 bp的开放阅读框,编码654个氨基酸,理论分子量和等电点分别为71.8 k Da和5.38;Pshsp90 c DNA序列包含2 172 bp的开放阅读框,编码724个氨基酸,理论分子量和等电点分别为83.5 k Da和4.93。Pshsp70和Pshsp70-4均含有Hsp70基因家族高度保守的基序,Pshsp70编码的氨基酸序列与烟粉虱Bemisia tabaci和家蚕Bombyx mori等昆虫的Hsp70的氨基酸序列一致性为85%;Pshsp70-4编码的氨基酸序列与白蜡蚧Ericerus pela和点蜂缘蝽Riptortus pedestris等昆虫的Hsp70的氨基酸序列一致性高达95%;Pshsp90也含有Hsp90基因家族高度保守的基序,Pshsp90编码的氨基酸序列与赤拟谷盗Tribolium castaneum和东亚小花蝽Orius sauteri等昆虫的Hsp90的氨基酸序列一致性为87%。热激蛋白基因表达量分析结果表明,在18℃恒温条件下,粉蚧2龄若虫的3个Ps Hsps基因的mRNA相对表达量均比对照（26℃）低,在32℃恒温条件下,各龄期的Hsp70基因的相对表达量均显著高于对照。在37~45℃下热激1 h并在26℃下恢复1 h,棉花粉蚧3个龄期的3个热激蛋白Ps Hsps基因的相对表达量随温度的升高总体呈增加趋势,相关性分析表明,除Pshsp70-4在雌成虫中的表达�[ Aim ] This study aims to study the roles of heat shock protein in the adaptation to temperature stress in Phenacoccus solenopsis. [ Methods ] Based on the successful transcriptome sequencing in P. solenopsis, we analyzed two sequences of heat shock protein 70 gene family [ PsHsp70 （GenBank accession no. KJ909505 ） and PshspTO-4 （GenBank accession no. KJ909506 ）] and one sequence of heat shock protein 90 gene family [ Pshsp90 （GenBank accession no. KJ909507 ）], and used comparative quantitative real-time PCR （RT-qPCR） to analyze the expression of genes PshspTO, PshspTO-4 and Pshsp90 at different developmental stages （2nd instar nymph, 3rd instar nymph and female adult） of P. solenopsis in response to different temperatures （ constant temperature 18 and 32℃, and temporary heat shock for 1 h at 37, 39, 41, 43 and 45℃ and then recovery for 1 h at 26℃ ） were detected. [ Results ] Sequence analysis showed that the PshspTO cDNA contains a 1 923 bp open reading frame, encoding a polypeptide of 641 amino acids with an estimated molecular mass of 70.9 kDa and an estimated isoelectric piont （pI） of 5.65. The PshspTO-4 cDNA contains a 1 962 bp open reading frame, encoding a polypeptide of 654 amino acids with an estimated molecular mass of 71. 8 kDa and an estimated isoeleetric piont （pI） of 5.38. The Pshsp90 eDNA contains a 2 172 bp open reading frame, encoding a polypeptide of 724 amino acids with an estimated molecular mass of 83. 5 kDa and an estimated isoelectric piont （pI） of 4.93. Both Pshsp70 and Pshsp70-4 contain the highly conserved functional motifs of the Hsp70 gene family. PsHsp70 shares 85% amino acid sequence identities with the Hsp70 proteins from Bemisia tabaci and Bombyx mori, while PsHsp70-4 shares 95% amino acid sequence identities with the Hsp70 proteins from Ericerus pela and Riptortus pedestris. Pshsp90 contains the highly conserved functional motifs of the Hsp90 gene family, and PsHsp90 shares 87% amino acid sequence identities with the Hsp90 proteins from T

关 键 词：棉花粉蚧 热激蛋白 温度胁迫 序列分析 mRNA相对表达量 实时荧光定量PCR 

分 类 号：Q968[生物学—昆虫学]