禾谷缢管蚜热激蛋白Hsp90基因的克隆和表达分析  

Molecular cloning and expression analysis of the Hsp90 gene in the bird cherry-oat aphid,Rhopalosiphum padi

作　　者：李玉婷[1] 赵奇[1] 房冰[1] 陈茂华[1] 

机构地区：[1]西北农林科技大学植物保护学院,农业部西北黄土高原作物有害生物综合治理重点实验室,陕西杨凌712100

出　　处：《植物保护学报》2017年第1期16-23,共8页Journal of Plant Protection

基　　金：国家自然科学基金(31272036,31471766);教育部高等学校博士学科点专项科研基金(20110204110001)

摘　　要：为探讨高温驯化对禾谷缢管蚜Rhopalosiphum padi Hsp90基因表达量的影响,以及其在不同发育阶段的表达,通过RT-PCR与RACE技术克隆了热激蛋白Hsp90基因的全长,命名为Rp Hsp90,采用生物信息方法分析了其序列特征,利用实时荧光定量PCR技术研究了Rp Hsp90在禾谷缢管蚜不同发育阶段和驯化后热激处理的表达量变化。结果显示:禾谷缢管蚜Rp Hsp90的c DNA全长为2 644 bp,编码727个氨基酸,分子量为83.2 k D。推导的氨基酸序列具有Hsp90蛋白家族的5个签名序列及C末端细胞质特征序列EEVD。系统进化树结果显示,Rp Hsp90与其它昆虫Hsp90具有很高的相似性。实时荧光定量PCR结果表明,不同发育阶段Rp Hsp90均有表达,并且4龄若蚜期表达量最高,显著高于其它龄期;27℃热驯化后禾谷缢管蚜Rp Hsp90表达量和常温24℃处理试虫无显著差异;热激处理显著诱导Rp Hsp90的表达,39℃热激处理Rp Hsp90表达量显著高于40℃热激处理;热驯化种群经热激处理后表达量显著高于常温热激处理。表明Rp Hsp90在禾谷缢管蚜不同龄期差异表达,且在该虫对热胁迫的响应中具有重要作用。To investigate the influence of heat acclimation on heat shock protein 90 in Rhopalosiphum padi and analyze the expression pattern of the gene at different developmental stages, the reverse transcription PCR（ RT-PCR） and rapid amplification of c DNA ends（ RACE） methods were used to amplify the full length c DNA of R. padi Hsp90 gene（ designated as Rp Hsp90）. Bioinformatic methods were used to analyze the characteristics of Rp Hsp90. The expression levels of the Rp Hsp90 gene at different developmental stages and under different heat treatments were detected by the real-time quantitative PCR. The results showed that the full-length c DNA of Rp Hsp90 was 2 644 bp,which encoded 727 amino acids,with a molecular weight of 83. 2 k D. The deduced amino acid sequence of Rp Hsp90 contained five signature sequences of Hsp90 family and a cytoplasmic character sequence（ EEVD） at C-terminal. The phylogenic tree indicated that Rp Hsp90 protein shared a high homology with Hsp90 proteins from other insect species. The real-time quantitative PCR analyses showed that the expression levels of Rp Hsp90 varied remarkably at different developmental stages. The mRNA level wasthe highest in the 4th-instar nymphs,which was significantly higher than that in other instars. No significant difference in expression level was found between 27℃ acclimation and the control at 24℃.The expression level of Rp Hsp90 was highly induced by heat shock treatments. The mRNA level of Rp Hsp90 under 39℃ treatment was significantly higher than that under 40℃ treatment. The population with 27℃ acclimation showed the highest expression level of Rp Hsp90 after heat shock. The current study indicated that the expression level of Rp Hsp90 varied at different developmental stages and played a role in the response of the pest to heat stress.

关 键 词：禾谷缢管蚜 HSP90 基因克隆 表达分析 

分 类 号：S433[农业科学—农业昆虫与害虫防治]