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【目的】昆虫在高温或农药的胁迫下,通过高效表达热休克蛋白(HSP)等建立应激自我保护机制。本研究为从转录组水平上认识大豆蚜Aphis glycines在热应激和吡虫啉胁迫下hsp70和hsc70 mRNA表达分子机制,进而寻找自我保护应激反应中的薄弱环节,为大豆蚜的生物防治提供理论基础。【方法】采用同源克隆、RACE技术和实时荧光定量PCR等方法研究不同热激时间和热激后不同恢复时间及不同吡虫啉浓度对大豆蚜4龄若虫hsp70和hsc70的表达影响。【结果】37℃热激后,大豆蚜4龄若虫中hsp70表达量先上调,1 h时升至对照组的10.36倍(P<0.05),然后逐渐下降。同样热激后恢复时间的长短对大豆蚜若蚜中hsp70的表达具有显著影响。热激处理后,大豆蚜若蚜中hsp70立即大量表达,表达量为对照组的8.78倍(P<0.05),随后表达量下降至对照组水平,而hsc70的表达量并没有显著变化(P>0.05)。大豆蚜若蚜受吡虫啉的胁迫时,其hsp70和hsc70的表达量受吡虫啉的浓度及胁迫的时间的影响,呈现先升高后下降的趋势,具有明显的短期效应。【结论】吡虫啉诱导大豆蚜hsp70和hsc70表达量的上调;而热胁迫对hsp70和hsc70 mRNA具有不同的表达模式,高温可以诱导hsp70的表达,但对hsc70没有明显的诱导作用。
【Objective】 Insects under high temperature or pesticide stress, through the expression of heat shock protein (HSP) to establish stress self-protection mechanism. In this study, we understand the molecular mechanism of the expression of hsp70 and hsc70 mRNA in soybean aphid Aphis glycines under heat stress and imidacloprid stress from the level of transcriptome, so as to find the weak link in the self-protection stress response and provide the theoretical basis for the biological control of soybean aphid . 【Method】 Homologous cloning, RACE and real-time fluorescence quantitative PCR were used to study the effects of different recovery time and different imidacloprid concentrations on the expression of hsp70 and hsc70 of 4th instar nymphs of soybean aphid. 【Results】 After heat shock at 37 ℃, the expression of hsp70 in the 4th instar nymphs of soybean aphid was up-regulated first, then increased to 10.36-fold (P <0.05) at 1 hour and then gradually decreased. The same recovery time after heat shock on soybean aphid hsp70 expression has a significant impact. After heat shock treatment, the expression level of hsp70 in soybean aphid was up to 8.78 folds (P <0.05), and then decreased to the level in control group, but the expression of hsc70 did not change significantly (P> 0.05). Soybean aphid, if treated by imidacloprid, the expression of hsp70 and hsc70 was increased firstly and then decreased with the concentration of imidacloprid and the time of stress, and had obvious short-term effect. 【Conclusion】 Imidacloprid induced the up-regulation of hsp70 and hsc70 in soybean aphids. However, heat stress had different expression patterns on hsp70 and hsc70 mRNA. High temperature induced the expression of hsp70 but had no obvious effect on hsc70.