禾谷缢管蚜热激蛋白Hsp90基因的克隆和表达分析

发布时间：2018-05-17 16:03

本文选题：禾谷缢管蚜 + Hsp　；参考：《植物保护学报》2017年01期

【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。
[Abstract]:In order to investigate the effect of high temperature acclimatization on the expression of Rhopalosiphum PADI Hsp90 gene of cereal constrictive aphid and its expression at different developmental stages, the whole length of the gene of heat shock protein Hsp90 was cloned by RT-PCR and RACE technology, named Rp Hsp90, and its sequence characteristics were analyzed by biological information method, and the real-time fluorescent quantitative PCR technology was used. The changes in the expression of Rp Hsp90 at different stages of development and heat shock after acclimation were investigated. The results showed that the total length of C DNA of the cereal constriction aphid Rp Hsp90 was 2644 BP, 727 amino acids encoded and the molecular weight of 83.2 K D. had 5 signature sequences of the Hsp90 protein family and the C terminal cytoplasmic sequence EEVD. The phylogenetic tree showed that Rp Hsp90 had high similarity with other insect Hsp90. Real time fluorescence quantitative PCR showed that Rp Hsp90 was expressed in different developmental stages, and the expression of aphid stage at 4 age was the highest and significantly higher than that of other ages. The expression of Rp Hsp90 in cereal constrictive aphid and at 24 C at normal temperature were not significant after thermal acclimation at 27. The expression of Rp Hsp90 was significantly induced by heat shock treatment, and the expression of Rp Hsp90 in heat shock treatment at 39 C was significantly higher than that of heat shock treatment at 40 C. The expression of heat acclimated population after heat shock treatment was significantly higher than that at normal temperature. It showed that Rp Hsp90 was expressed in different ages of cereal constriction aphid, and it was important in the response of the insect to heat stress.
【作者单位】：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;
【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)
【分类号】：S433

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