Sun Yang, Sheng Yang, Bai Lixin, Zhang Yongjun, Xiao Yingfang, Xiao Liubin, Tan Yongan, Shen Youmi
Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, China.
Cell Stress Chaperones. 2014 Sep;19(5):725-39. doi: 10.1007/s12192-014-0500-0. Epub 2014 Mar 13.
In this study, we cloned a full-length cDNA of heat shock protein (HSP) gene of Apolygus lucorum (Meyer-Dür) [AlHSP90, KC109781] and investigated its expression in response to temperature and pesticide stresses. The open reading frame (ORF) of AlHSP90 is 2,169 bp in length, encoding a 722 amino acid polypeptide with a predicted molecular weight of 82.99 kDa. Transcriptional and translational expression profiles of AlHSP90 under extreme temperature or pesticide stresses were examined by fluorescent real-time quantitative PCR and Western blot. Results showed that the expression profiles of AlHSP90 protein were in high agreement with those of AlHSP90 RNA and indicated that AlHSP90 was not only an important gene for A. lucorum adults in response to extremely high temperature, but also involved in the resistance or tolerance to cyhalothrin, imidacloprid, chlorpyrifos, and emamectin benzoate, especially for female adults to emamectin benzoate and for male adults to cyhalothrin. Transcriptional results of AlHSP90 also confirmed that AlHSP90 was an important gene involved in the resistance or tolerance to both temperature and pesticide stresses. In addition, our study also revealed that ∼24 °C may be the suitable temperature range for A. lucorum survival, which is also confirmed by the results of the expression of AlHSP90, the nymph mortality, and the intrinsic rate of increase (r m) when A. lucorum is reared at six different temperatures. Therefore, these studies are significant in elucidating the AlHSP90 in response to temperature and pesticide stresses and would provide guidance for A. lucorum management with different pesticides or temperatures in fields.
在本研究中，我们克隆了绿盲蝽（Apolygus lucorum (Meyer-Dür)）热休克蛋白（HSP）基因的全长cDNA [AlHSP90，登录号KC109781]，并研究了其在温度和农药胁迫下的表达情况。AlHSP90的开放阅读框（ORF）长度为2169 bp，编码一个由722个氨基酸组成的多肽，预测分子量为82.99 kDa。通过荧光实时定量PCR和蛋白质免疫印迹法检测了AlHSP90在极端温度或农药胁迫下的转录和翻译表达谱。结果表明，AlHSP90蛋白的表达谱与AlHSP90 RNA的表达谱高度一致，表明AlHSP90不仅是绿盲蝽成虫应对极端高温的重要基因，还参与了对高效氯氟氰菊酯、吡虫啉、毒死蜱和甲氨基阿维菌素苯甲酸盐的抗性或耐受性，尤其是雌虫对甲氨基阿维菌素苯甲酸盐和雄虫对高效氯氟氰菊酯的抗性或耐受性。AlHSP90的转录结果也证实，AlHSP90是参与温度和农药胁迫抗性或耐受性的重要基因。此外，我们的研究还表明，约24℃可能是绿盲蝽生存的适宜温度范围，这也通过在六个不同温度下饲养绿盲蝽时AlHSP90的表达、若虫死亡率和内禀增长率（rm）的结果得到了证实。因此，这些研究对于阐明AlHSP90对温度和农药胁迫的响应具有重要意义，并将为田间不同农药或温度条件下的绿盲蝽防治提供指导。
