亚洲玉米螟交变温度下热休克蛋白基因(HSPs)的表达分析

摘要

Hsc70 and Hsp90 are considered indicators related to ability of insect resistance to environmental stress and adaption to temperature, in order to detect the influence of circadian temperature on the cold resistances of diapause larvae of Asian corn borer, Ostrinia furnacalis(Guenée). The diapause larvae were treated by alternating temperature, -20℃ induction for 6 hours and 4℃ recovery for 18 hours. On different times, the super-cooling points were tested, and the relative expression of Hsc70 and Hsp90 gene were detected using qPCR, and with analyzed by SPSS. Hsc70 and Hsp90 genes were found in response to alternating temperature, the expressions decreased in low temperature induced phase, and maintained in a lower expression level (induction stage 0~3h). Until induced 6h, the expression level began to rise. 4℃ recovery treatment followed. At first the expressions decreased, however, they increased after recovery 1 hour and reached maximum after recovery 3 hours. The results showed that Hsc70 and Hsp90 were temperature sensitive proteins, and their expression was significantly affected by alternating temperature, and the low temperature was conducive to the low expression level. The correlation between the expression of HSPs and the change of super-cooling point was analyzed. The change of Hsc70 expression was significantly correlated with the change of super-cooling point, Hsp90 was not significant. These results suggest that Hsc70 may play an important role in cold resistance of Asian corn borer.%热休克蛋白基因(heat shock proteins,HSPs)Hsc70及Hsp90是目前认为与昆虫抵御环境压力及温度适应有关的指标,为了检测较大日温差对亚洲玉米螟滞育幼虫抗寒性的影响,本研究模拟东北地区昼夜温差效应,对亚洲玉米螟滞育幼虫进行6h的-20℃低温诱导和随后18h的4℃恢复处理,在不同的时间点测定其过冷却点,使用实时荧光定量PCR(qRT-PCR)技术检测Hsc70及Hsp90的相对表达量,并利用SPSS对最终的表达量进行统计学分析,发现Hsc70及Hsp90在低温诱导阶段的表达出现下调,且维持在一个较低的表达水平(低温诱导0~3h).直至低温诱导6h,其表达量开始回升,此时,进行4℃恢复处理,发现起初恢复阶段Hsc70及Hsp90的表达明显下调,而在恢复1h后2种热休克蛋白表达量逐渐增加,恢复3h时达到表达量的峰值,而后又逐渐下降.此结果说明,Hsc70及Hsp90是温度敏感性蛋白,其表达量受交变温度的影响显著,并且低温有利于其表达量维持在较低水平.对Hsc70及Hsp90表达量变化规律与过冷却点变化规律进行相关性分析可知,Hsc70表达量的变化规律与过冷却点变化规律显著相关,Hsp90则相关不显著.此结果表明Hsc70可能对亚洲玉米螟抗寒性起到了重要作用.