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为了检测B型烟粉虱和温室白粉虱在受到温度胁迫时热激基因Hsp70的差异表达谱,克隆了2种粉虱Hsp70基因的全长cDNA序列,并用实时定量PCR的方法分析了在不同温度条件下该基因的表达谱。结果表明:B型烟粉虱和温室白粉虱的Hsp70基因BTH-sp70和TVHsp70(GenBank登录号分别为DQ093385和EU861391),在5’-非编码区(UTR)中有类似TATA-box样元件;在3’-UTR中有poly(A)信号AATAA和富含A-T区;根据基因开放阅读框序列(ORF)推导的氨基酸序列均含有全部3个Hsp70蛋白家族高度保守的基序。在整个检测的温度范围内(-19℃～46℃),诱导温室白粉虱Hsp70表达的起始温度(Ton)或最大温度(Tmax)要比B型烟粉虱低2.5℃～7.5℃。这些结果表明,所克隆的基因属于有功能的温度诱导型Hsp70基因;在基因表达水平上,温室白粉虱比B型烟粉虱更耐冷,而后者更耐热;Hsp70的Ton(或者Tmax)能代表这2种粉虱的温度耐受能力。本研究结果在一定程度上解释了自然界中这2种粉虱种群地理分布和季节发生差异的原因。
In order to detect the differential expression profiles of heat shock gene Hsp70 in B. tabaci B and B. tabaci during temperature stress, the full-length cDNA sequences of Hsp70 genes in B. tabaci were cloned and analyzed by real-time quantitative PCR at different temperatures Under the conditions of the gene expression profile. The results showed that Hsp70 genes BTH-sp70 and TVHsp70 (GenBank accession numbers DQ093385 and EU861391, respectively) showed similar TATA-box-like elements in the 5’-UTR of B. tabaci B and B. tabaci B. tabaci BTH- The A (A) signal and the AT region are rich in 3’-UTR. The amino acid sequences deduced from the open reading frame (ORF) of the gene all contain highly conserved motifs of all 3 Hsp70 protein families. The starting temperature (Ton) or maximum temperature (Tmax) of Hsp70 expression in B. tabaci was 2.5 ℃ ~ 7.5 ℃ lower than that of B. tabaci in the whole temperature range (-19 ℃ ~ 46 ℃). These results indicate that the cloned gene belongs to a functional temperature-inducible Hsp70 gene. At the gene expression level, B. tabaci are more resistant to cold than B. tabaci B, while the latter is more thermotolerant. The Ton (or Tmax) energy of Hsp70 Represents the temperature tolerance of these two whiteflies. The results of this study to some extent explain the nature of the two whitefly population geographical distribution and seasonal differences in the reasons.