| 摘要: |
| 【目的】对金纹细蛾Hsp90基因进行生物信息学和表达模式分析，为深入研究金纹细蛾PrHsp90与生长发育和抗逆行为的关系奠定基础。【方法】利用分子克隆技术获得金纹细蛾Hsp90基因片段，并对其进行生物信息学分析。同时利用RT-qPCR技术检测Hsp90在金纹细蛾不同组织、不同生长发育阶段和高温胁迫后的表达情况。【结果】获得了金纹细蛾Hsp90基因的一条长度为942 bp的片段序列，命名为PrHsp90，登录号为KP671600。序列分析显示，其对应的氨基酸序列与其他昆虫的相似性很高，与棉铃虫（ADP37710.1）、斜纹夜蛾（ADK55516.1）、烟夜蛾（ADM26742.1）等鳞翅目昆虫的同源性在98%以上，表明Hsp90基因家族具有高度保守性。实时荧光定量结果表明，PrHsp90在金纹细蛾翅中的相对表达量显著高于头、胸、腹和足；金纹细蛾各个生长发育阶段都有PrHsp90表达，但其在成虫体内的相对表达量显著高于蛹期和幼虫期。高温胁迫下PrHsp90的表达水平随着温度的升高而升高，且经过35，38，41和44 ℃处理后的金纹细蛾，其体内PrHsp90相对表达量均显著提高，约为对照处理表达量的3～15倍。【结论】金纹细蛾PrHsp90的表达具有组织特异性，且在其生长发育的各个阶段和温度胁迫中均可能起重要作用。 |
| 关键词: 金纹细蛾 热激蛋白 生长发育 高温胁迫 |
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| 基金项目:陕西省农业厅重点农业科技示范推广项目（ZDKJ-2014-32） |
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| Cloning and expression analysis of Hsp90 gene fragment from Phyllonorycter ringoniella (Lepidoptera:Gracilariidae) |
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WANG Guanhua,LI Xin,GUO Changning,et al
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| Abstract: |
| 【Objective】The fragment of Hsp90 gene from Phyllonorycter ringoniella was cloned and its biological information and expression patterns under different treatments were analyzed to provide basis for further study about the relationships between PrHsp90 and the development and heat resistance of P.ringoniella.【Method】The Hsp90 fragment was cloned using RACE-PCR technology.The expression patterns of Hsp90 in different tissues at developmental stages under different heat treatments were analyzed using RT-qPCR.【Result】The Hsp90 gene fragment with length of 942 bp was obtained,which was named PrHsp90 and the access number in GenBank was KP671600.Sequence analysis showed that its amino acid sequence shared high homology with Hsp90 from other insects.It had 98% and higher homology with Hsp90 from Heliothis armigera (ADP37710.1),Spodoptera litura (ADK55516.1) and Helicoverpa assulta (ADM26742.1).Expression analysis of PrHsp90 in different tissues indicated its expression level was significantly higher in wing than in head,thorax,abdomen and foot.The PrHsp90 transcript was detectable at all developmental stages with dramatically higher expression in adults than in larvae and pupae.Under heat treatments,the expression of PrHsp90 increased with the increase of temperature.After heat treatments at 35,38,41 and 44 ℃,its expression increased significantly to 3－15 times of that in control.【Conclusion】Expression of PrHsp90 is tissue-specified and it plays key roles in development and heat resistance of P.ringoniella. |
| Key words: Phyllonorycter ringoniella HSP developmental stages thermal stress |
