摘要
采用RT-PCR及RACE技术克隆了三叶斑潜蝇Hsp90基因全长cDNA序列,并用实时定量RT-PCR的方法检测其在不同发育阶段受到高温胁迫后的表达水平。该基因的cDNA序列全长2 408bp,开放阅读框为2 145bp,编码714个氨基酸;5′非编码区为151bp,3′非编码区为112bp。该基因推导的氨基酸序列与其他昆虫同源序列比较有很高的相似性(80%～99%)。聚类分析结果显示三叶斑潜蝇与美洲斑潜蝇和南美斑潜蝇的亲缘关系最近。实时荧光定量PCR检测结果表明三叶斑潜蝇Hsp90基因的表达受到热胁迫的诱导,诱导3龄幼虫最大表达量的温度比诱导其他发育阶段的温度低,在43℃时预蛹和蛹的表达量在整个生命周期中最高,在检测的高温胁迫条件下,雄虫比雌虫的表达量更高。该结果为阐明三叶斑潜蝇胁迫耐受能力及其对其他潜蝇种群的取代机制奠定了分子基础。
A full-length cDNA encoding heat shock protein 90 （Hsp90） was cloned from Liriomyza trifolii using RT-PCR and RACE methods, and the expression levels of this gene in relation to high temperature stress during different developmental stages were investigated using real-time quantitative PCR （qRT-PCR）. The cDNA is 2 408 bp in length, containing an open reading frame （ORF） of 2 145 bp, encoding 714 amino acid residues, with a 5runtranslated region （UTR） of 151 bp and a 31UTR of 112 bp. Cluster analysis confirmed that the deduced amino acid sequence shared high identity with the reported homologous sequences from other insects （80% 99%）. Evolution results showed the genetic relation of L. trifolii was more close to that of L. salivae and L. huidobrensis, qRT-PCR analysis showed that expression of Hsp90 gene could be induced by heat stress. The maximum of inductive temperature of the third instar larvae was lower than that of other developmental stages. Gene expression attained the maximum at the stage of prepupa and pupa at 43℃. Higher expression level was ob- served in male than in female when exposure to heat stress tested. Our research provided a way forward for further research on stress tolerance of L. trifolii and the displacement mechanism of this insect to other leafminer populations.
出处
《植物保护》
CAS
CSCD
北大核心
2013年第5期110-116,122,共8页
Plant Protection
基金
北京市自然科学基金(6122026)
北方民族大学自主科研基金研究生创新项目(2012XYC046)
