| 李鸿波，戴长庚，胡阳,2023,黏虫HSP90/70组织蛋白基因的分子特征与表达特性[J].环境昆虫学报,45(1):114-123 |
| 黏虫HSP90/70组织蛋白基因的分子特征与表达特性 |
| Molecular and expression characterization of an HSP90/70 organizing protein(HOP)gene in the Mythimna separata (Lepidoptera: Noctuidae) |
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| DOI： |
| 中文关键词: 黏虫 HSP90/70组织蛋白 基因克隆 表达特性 |
| 英文关键词:Mythimna separata heat shock 90/70 organizing protein gene cloning expression characterization |
| 基金项目:贵州省基础研究计划（黔科合基础[2020]IZ022） |
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| 中文摘要: |
| 为解析黏虫Mythimna separata响应环境胁迫的分子机制，采用RACE和RT-PCR的方法从黏虫中克隆了 1个 HSP90/70组织蛋白(heat shock 90/70 organizing protein, HOP)基因，并采用qRT-PCR分析了其在多种生物和非生物胁迫下的表达特性。结果表明，MsHOP具有1个1 626 bp的开放阅读框，编码541个氨基酸，预测的分子量为61.7 kDa。序列分析表明，MsHOP主要由已报道的3个保守四肽重复结构域（TPR1, TPR2A和TPR2B）和2个天冬氨酸-脯氨酸重复基序（DP）构成。基于鳞翅目昆虫HOP构建的系统进化树显示MsHOP与棉铃虫Helicoverpa armigera HOP和烟芽夜蛾Heliothis virescens HOP的亲缘关系最近。qRT-PCR分析表明，MsHOP在黏虫所有发育时期和组织均表达，分别在5龄幼虫和雌成虫中肠的表达量最高。在30℃下处理1 h后，MsHOP的表达水平较对照显著提高，而在33℃~39℃下处理1 h后MsHOP的表达水平则与对照差异不显著。饥饿24~72 h后，MsHOP的表达与对照相比无明显变化。Bt能显著诱导MsHOP的表达，其表达水平随Bt浓度的升高而上调，并在64 IU/mL浓度下的表达量最高。幼虫饲养密度对MsHOP的表达也具有显著影响，其表达水平随密度的升高而上调，在20 头/瓶的饲养密度下达到最大值。这些结果表明，MsHOP可能在黏虫的生长发育、抵御杀虫剂和密度胁迫中发挥重要作用。 |
| 英文摘要: |
| To analyze the responsive mechanism of Mythimna separata to environmental stress, a heat shock 90/70 organizing protein (HOP)homolog gene was identified by RACE and RT-PCR methods, and its expression characterization under multiple biotic and abiotic stressors was also examined by qRT-PCR. Results showed that the identified MsHOP had a 1 626 bp ORF encoding a protein of 552 amino acids with a calculated molecular mass of approximately 61.7 kDa. Its deduced amino acids comprised of three conserved tetratricopeptide repeated domains (TPR1, TPR2A and TPR2B)and two aspartic acid-proline (DP)repeat motifs as reported in other insects. Phylogenetic tree based on Lepidoptera HOPs showed that MsHOP was closely related to Helicoverpa armigera HOP and Helicoverpa virescens HOP. qRT-PCR analysis showed that MsHOP was expressed in various developmental stages and tissues of the fourth-day-old female adult, and exhibited the highest expression level in the 5th instar larval stage and midgut, respectively. MsHOP was significantly up-regulated after exposure to 30℃ for 1 h, but its expression level showed no difference at 33℃ or higher temperature as compared with that of the control (25℃). MsHOP was not changed after starvation for 24~72 h. After feeding on artificial an diet treated with Bt, MsHOP expression was significantly up-regulated and showed the highest transcription level at 64 IU/mL. The expression level of MsHOP was significantly induced with increasing larval density, and showed the highest expression at 20 larvae per jar. Taken together, the full-length cDNA of MsHOP was successfully obtained, and it might play important role in development, Bt and crowding stress response in M. separata. |
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