›› 2014, Vol. 57 ›› Issue (7): 777-786.

• 研究论文 • 上一篇    下一篇

灰飞虱热激蛋白Hsp90基因的克隆、序列分析与表达模式

张青1,2, 陆明星1, 祝树德1,*   

  1. (1. 扬州大学园艺与植物保护学院, 江苏扬州 225009; 2. 苏州市农业科学院, 江苏苏州 215155)
  • 出版日期:2014-07-20 发布日期:2014-07-20

Cloning, sequence analysis and expression profiling of heat shock protein 90 gene in the small brown plant hopper, Laodelphax striatellus (Hemiptera: Delphacidae)


ZHANG Qing1,2, LU Ming-Xing1, ZHU Shu-De1,*   

  1. (1. School of Horticulture and Plant Protection, Yangzhou University, Yangzhou, Jiangsu 225009, China; 2. Suzhou Academy of Agriculture Sciences, Suzhou, Jiangsu 215155, China)
  • Online:2014-07-20 Published:2014-07-20

摘要: 【目的】灰飞虱Laodelphax striatellus (Fallén)是水稻上的重要害虫,它严重影响了水稻的产量和品质,特别由其传播的水稻条纹叶枯病毒对水稻的危害甚至更为严重。灰飞虱分布广,对环境有很强的适应性。本研究旨在探讨灰飞虱对温度胁迫适应的分子机制进行了初步的探讨。【方法】本研究采用RT-PCR与RACE技术克隆了热激蛋白Hsp90基因的全长,利用各种生物信息方法分析了该热激蛋白的特征,实时荧光定量PCR技术用于研究Hsp90基因在不同生长发育阶段和不同温度条件下表达变化规律。【结果】我们获得一条Hsp90 基因的cDNA全长序列,命名为LsHsp90(GenBank登录号为KF660250)。 LsHsp90全长为2 740 bp,编码729个氨基酸,等电点为5.0,分子量为83.7 kDa。氨基酸序列中含有 Hsp90蛋白家族的5个签名序列及胞质特征序列 MEEVD。结构预测表明LsHsp90具有Hsp90蛋白家族典型的结构特征。系统进化关系分析表明它与多种昆虫的Hsp90序列有较高的同源性。不同发育阶段的灰飞虱体内LsHsp90表达水平处于动态的变化过程中,并在4龄若虫期达到最大值,最低表达量在其雌成虫阶段。不同性别的灰飞虱成虫体内LsHsp90表达量有显著差异(P =0.008)。高温和低温胁迫都可以诱导灰飞虱体内LsHsp90的表达,最大的表达量分别在 40℃和-9℃。在40℃下处理不同时间后发现,LsHsp90的表达水平在处理后0.5 h时最高,但随着时间的延长,LsHsp90的表达水平逐渐降低。而在-4℃下,LsHsp90的表达水平在处理后1 h时达到最大值。【结论】灰飞虱体内的LsHsp90可以响应温度的诱导,它可能在灰飞虱温度胁迫适应过程中起到重要的作用。

关键词:

Abstract:  【Aim】 The small brown planthopper Laodelphax striatellus (Fallén), an important rice pest insect, feeds on rice and causes the decrease of yield and quality. Especially the rice stripe virus transmitted by L. striatellus induces more serious loss of rice. The small brown plant hopper is widely distributed and adapts to various environments. This study aims to clarify the molecular adaption mechanisms of L. striatellus under temperature stress preliminarily. 【Methods】 A full-length cDNA encoding Hsp90 from L. striatellus was cloned using RT-PCR and RACE technique. Different bioinformatics methods were used to analyze the characteristics of Hsp90. The expression levels of the Hsp90 gene among different developmental stages and under different temperatures were detected by the real-time PCR. 【Results】 The complete cDNA of LsHsp90 deposited in GenBank accession no. KF660250 is 2 740 bp in length, which encodes a protein of 729 amino acids, with the theoretical isoelectric point of 5.0 and molecular weight of 83.7 kDa. The amino acid sequence contains five signature sequences of Hsp90 family and a C-terminal cytoplasmic character sequence (EEVD). The LsHsp90 carries a classic Hsp90 family structural signature. The phylogenic tree demonstrated that LsHsp90 has high homology with Hsp90 proteins from some other insect species. The real-time PCR analyses exhibited that the expression levels of LsHsp90 varied remarkably in different developmental stages. The mRNA level observed was the highest in the 4th instar nymphs of L. striatellus, the lowest in female adults, and showed significant differences between male and female adults (P=0.008). The LsHsp90 mRNA level of L. striatellus could be induced by heat and cold temperature stress. The highest LsHsp90 mRNA level was observed at 40℃ and -9℃, respectively. At 40℃, LsHsp90 mRNA level reached a peak at 0.5 h after treatment, and then decreased with treatment time. However, at -4℃, the peak of LsHsp90 mRNA level was observed at 1 h after treatment. 【Conclusion】 The LsHsp90 gene in L. striatellus responds to temperature stress, and it may play an important role in the adaption process of the pest to temperature stress.

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