As a member of the family of small heat shock proteins, it is possible that the sHsp21 can be induced under certain circumstances, for example, under the pressure from external environment. As the heat shock is able to adjust the folding of protein and protect cells from damage of the outside stress. Focusing on the sHsp21, this paper mainly studied the relevant genes of Agasicles hygrophila Selman & Vogt, which is a promising biological control agent for invasive Alternanthera philoxeroides and carries on the expression of heat shock induced, as well as total length of amplification by RACE. The full length we observed of cDNA is 693 bp while it contains a complete set of open reading frame of 567 bp, encoding 188 amino acids. Using constant temperature 30℃ as control. We manged to measure the sHsp21 expressive quantity of Hunan population and Hainan population under the simulative high-temperature period (10∶00-14∶00)of summer at 36℃、39℃ respectively. The consistant heat shocking lasted six consecutive days and sampling is arranged to be done 4 hours afterwards. The result of Real-time fluorescent quantitative PCR analysis suggests that: Hunan population under the 39℃, 36℃ high temperature induced sHsp21 gene expressive quantity is significantly higher than the control group 30℃; Hainan population 36℃ under the high temperature induced the sHsp21 expression of the quantity is of no significant difference to the control group 30℃. Meanwhile, under the 39℃ high temperature sHsp21 gene expressive quantity is significantly higher than the 36℃ and 30℃. Which leads to the conclusion that sHsp21 on Agasicles hygrophila high temperature tolerance may play an important role as well as the high expression of sHsp21 may cause negative effect to the insect (on its own), therefore provided us a certain foundation as we delve one step closer into the biological function of sHsp21.%sHsp21(小热激蛋白)是小热激蛋白家族的成员,在外界环境压力下能够被诱导,热激后能够调节蛋白质的折叠和保护细胞免受外界胁迫.本文研究了入侵杂草空心莲子草Alternanthera philoxeroides生防天敌莲草直胸跳甲的一个关键基因sHsp21,并对其进行热激诱导表达以及RACE全长扩增.获得全长cDNA 693 bp并完整的开放阅读框567 bp,共编码188个氨基酸.以30℃为对照,分别测定莲草直胸跳甲Agasicles hygrophila Selman & Vogt长沙种群和海南种群在模拟夏季特定10∶00-14∶00时段36℃、39℃高温条件下sHsp21的表达量.连续热激 6 d,每天4 h热激后取样,实时荧光定量PCR分析显示:湖南种群高温39℃和36℃诱导下sHsp21表达量都显著高于对照组30℃;海南种群36℃诱导下的表达量与对照组30℃差异不显著,39℃诱导下的表达量显著高于对照组30℃和36℃.结果表明,sHsp21在湖南和海南两地理种群不同耐热能力上起到重要作用,并且sHsp21的高表达可能对其虫体产生副作用,为下一步深入探究其生物学功能奠定了一定的基础.
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