短时高温胁迫对褐飞虱存活及其热激蛋白基因hsp70表达的影响  

Effects of brief heat shock on the survival and hsp70 expression of brown planthopper Nilaparvata lugens( Stl)

作　　者：季璐[1] 崔旭红[1] 朱敏[1] 

机构地区：[1]中国计量学院生命科学学院,浙江省生物计量及检验检疫技术重点实验室,杭州310018

出　　处：《植物保护学报》2015年第4期619-625,共7页Journal of Plant Protection

基　　金：国家"973"计划(2010CB126200);国家公益性行业(农业)科研专项(200903051);浙江省自然科学基金(LQ12C14005)

摘　　要：为探讨短时高温胁迫对褐飞虱成虫存活的影响及热激蛋白基因hsp70在此过程中的作用,通过室内试验观察羽化24 h内的褐飞虱成虫经过不同高温（31~40℃）暴露和不同恢复时间（1~72 h）后的存活及其体内hsp70表达情况,并构建了褐飞虱hsp70实时荧光定量RT-PCR检测体系。结果表明,褐飞虱在31~39℃下暴露2 h对其成虫存活无显著影响,39.5℃下其存活率显著降低,为44.4%,40℃下无成虫存活;在37℃下暴露2 h后,在12 h的恢复时间内褐飞虱的存活率与对照组无显著差异,但随着恢复时间延长存活率迅速下降。缓和高温（31℃和33℃）对褐飞虱成虫hsp70表达具有诱导作用,其表达量分别为对照组的3.8和6.7倍;在35℃和37℃时达到最高峰,为对照组的291.2和232.5倍;当温度达到39℃和39.5℃时hsp70表达量呈下降趋势,但仍为对照组的46.7和26.2倍。褐飞虱37℃处理2 h后,随着恢复时间的延长,hsp70基因表达水平迅速下降,恢复48、72 h时,仅为对照组的0.4和0.7倍。推测该基因的表达在褐飞虱抵抗短时高温胁迫过程中具有重要作用。To study how Nilaparvata lugens（ Stl） may adapt to ecologically relevant levels of heat stress,the survival rates and the expression levels of heat shock protein gene hsp70 of the adults emerged within 24 h and briefly（ 2 h） exposed to high temperatures were determined. A real-time fluorescence quantitative RT-PCR method was developed to quantify the hsp70 expression levels in N. lugens exposed to 31- 40℃for 2 h,or recovered for 1- 72 h after a 2-hour-heat shock exposure at 37℃. The results showed that,within the temperature range of 31- 39℃,the adult survival rates were not significantly different between the heat shock treatment groups and the control. However,when the heat shock temperature was raised to39. 5℃,its adult survival rate was significantly lowered to 44. 4%,and no adults survived at 40℃ for 2 h.The survival rate of the group with 12-hour-recovery after a 2-hour-heat shock at 37℃ was not significantly different from the control. However,it decreased rapidly with the increasing recovery time. Meanwhile,a c DNA fragment encoding the putative HSP70 was cloned which comprised 797 bp（ corresponding to 265 amino acid residues） and contained two signature sequences of the HSP70 family. The quantitative RTPCR experiments showed that a mild heat shock could induce hsp70 expression in N. lugens. The hsp70 expression levels at 31℃,33℃,35℃ and 37℃ were 3. 8,6. 7,291. 2 and 232. 5 times higher than the control,respectively,and then decreased rapidly to 46. 7 and 26. 2 times at 39℃ and 39. 5℃. With the extension of the recovery time,the hsp70 expression level decreased quickly,and was down to 0. 4 and 0. 7times of the control after 48 and 72 h recovery,respectively. The expression of hsp70 may be critical for N.lugens to resist and adapt to the short-time heat-shock stresses.

关 键 词：褐飞虱 高温胁迫 存活率 热激蛋白基因hsp70 

分 类 号：S435.112.3[农业科学—农业昆虫与害虫防治]