橡胶树HbHsfB4b基因的克隆、组织特异性表达及亚细胞定位分析

Cloning,Tissue-specific Expression,and Subcellular Localization Analysis of HbHsfB4b Gene in Rubber Tree

热激转录因子(heat shock transcription factor,Hsf)在植物组织发育、结构发生和非生物胁迫响应等方面具有重要的调控作用。为了研究Hsf在橡胶树抵御逆境中的调控功能,本研究基于橡胶树基因组数据库克隆橡胶树HbHsfB4b基因全长开放阅读框序列,通过构建瞬时表达载体,明确亚细胞定位,采用RT-qPCR技术检测其在橡胶树不同发育组织中的表达量。结果显示,HbHsfB4b基因全长开放阅读框为1080 bp,编码359个氨基酸,分子量为40.25 kD,理论等电点为7.28,亲水性蛋白,该蛋白由3个α-螺旋和4个β-折叠组成。序列比对分析显示,橡胶树HbHsfB4b蛋白与麻风树JcHsfB4和蓖麻RcHsfB4蛋白相似性较高,分别为91.99%和87.98%。RT-qPCR分析结果显示,HbHsfB4b在古铜期叶片中表达量最高。HbHsfB4b蛋白定位于细胞核。本试验为进一步研究HbHsfB4b基因在橡胶树中的生物学功能提供参考。

Heat shock transcription factor(Hsf)plays an important regulatory role in plant tissue development,structural development,and abiotic stress response.To investigate the regulatory function of Hsf in resisting adversity.In this study,the full-length open reading frame sequence of HbHsfB4b gene was cloned based on the rubber tree genome database,transient expression vectors were constructed to clarify the subcellular localization,detection its expression in different developing tissues of rubber trees using RT-qPCR.The results showed that the full-length open reading frame of HbHsfB4b gene was 1080 bp,encoded 359 amino acids,the molecular weight was 40.25 kD and theoretical isoelectric point was 7.28,hydrophilic protein,which consisted of threeα-helix and fourβ-sheet.Sequence alignment analysis showed that the HbHsfB4b protein had high similarity with JcHsfB4(Jatropha curcas)and RcHsfB4(Ricinus communis)proteins,with 91.99%and 87.98%,respectively.The RT-qPCR analysis revealed that the HbHsfB4b gene was most abundantly expressed in the copper-drown stage leaves.This study provides a reference for further research on the biological function of HbHsfB4b gene in rubber trees.