不同温度影响下绿盲蝽丝氨酸蛋白酶基因AlSP3的表达谱分析

Expression Patterns of the Serine Protease Gene AlSP 3 in Apolygus lucorum(Hemiptera:Miridae) at Different Temperatures

利用荧光定量PCR技术，系统分析了不同温度处理下绿盲蝽成、若虫丝氨酸蛋白酶基因A1SP3的表达谱。结果表明：在24℃下，若虫各虫龄AISP3表达量差异未达到显著水平fP〉0．05）．而雌虫交配前期与雄虫交配后期的A1SP3表达量分别达l龄若虫对照的19．71倍与7．16倍，显著高于其他时期。但在21～30℃内，4～5龄若虫AISP3的表达量随温度升高而上升，2～5龄若虫在30℃时AISP3的表达量均达最高；不同温度处理下，刚羽化、交配前的雌、雄成虫的A1SP3的表达量较24℃对照均显著升高（P〈0．01）；而交配后的雌、雄成虫的表达量却是18℃下显著高于其他温度处理。此外，极端温度4℃与40℃诱导后，雌、雄成虫AISP3的表达量均显著高于24℃。绿盲蝽AISP3的表达谱综合分析表明，外在环境温度与绿盲蝽虫龄与性别因素均可显著影响绿盲蝽A1SP3的表达量（P〈0．01），并存在一定的互作效应。由此可见，AJSP3对于绿盲蝽雌成虫早期获取寄主营养具有重要作用，并且温度是影响AISP3表达的一个很重要的因素。

As an important digestive enzyme gene, the serine protease gene AISP 3 plays a key role in Apolygus lucorum for di- gesting conventional cotton. To understand the effect of different temperatures on the expression level of A1SP 3, we analyzed the expression patterns ofAISP 3 in A. lucorum reared_at different temperatures using real-time polymerase chain reaction. The results showed that at the normal temperature of 24 ℃, the expression of AISP 3 in A .lucorum at different nymphal stages was similar. The expressions of AISP 3 in the female adult at the pre-mating stage and the male adult at post-mating stage were 19.71- and 7.16-fold higher than their control at the 1 a nymphal stage, respectively, which were significantly higher than at any other stages. However, expression in the 4# to 5# A. lucorum at the nymphal stage increased with rising temperature in the range of 21-30 ℃. At 30 ℃, the expression ofA1SP 3 in the 2nd to 5th A. lucorum was the highest. The expression levels of A1SP 3 in adult A. lucorum reared at different temperatures during the new emergence and pre-mating stages were significantly higher than that of the control reared at 24 ℃ （P〈 0.01）, while the expression during the post-mating stage at 18℃ was higher than that at other temperatures. When incubated at extreme temperatures （4 ℃and 40 ℃）, the expression of AISP3 in female and male adult A. lucorum was significantly higher than that at 24 ℃ （P 〈 0.01）. Taken together, our results showed that the external envi- ronment temperature and the internal stage ofA. lucorum or its sex all had significant effects on the expression ofAISP 3 in A. lu- corum （P 〈 0.01）, and that there was some interactions between them. Therefore, AISP 3 is very important for the female adult A. lucorum to obtain nutrition at the early stages and temperature is an important limiting factor for the expression of AISP 3.