摘要
热激转录因子(HSF)基因是植物热胁迫响应的重要转录调节基因,在植物胁迫应答和其他抗逆反应过程中起着关键作用。为研究大蒜热激反应的分子机制,本研究基于大蒜转录组数据,以徐蒜6号为试验材料,采用同源克隆方法获得编码HSF的AsHSFB1基因。序列分析结果显示,AsHSFB1含有882 bp的开放阅读框,编码293个氨基酸,其蛋白质含有热激转录因子的特征结构域。在进化关系上,AsHSFB1与拟南芥AT4G11660(AtHSFB2B)同源性最高。亚细胞定位结果表明,AsHSFB1蛋白主要定位在细胞核和细胞质上。本研究采用同源重组技术成功构建pCAMBIA1305-AsHSFB1过表达载体,为进一步研究该转录因子基因的功能,培育耐热大蒜品种奠定基础。RT-PCR分析结果表明,不同大蒜品种中,AsHSFB1基因在叶片中相对表达水平均最高,具有组织表达特异性;38℃高温胁迫处理下,徐蒜6号中的AsHSFB1基因相对表达水平在24 h时明显上调;4℃低温胁迫下,徐蒜815和徐蒜6号中AsHSFB1基因相对表达水平的变化趋势相似,均先上升后下降,在处理4 h时相对表达水平达到峰值。
Heat shock transcription factor(HSF)is an important transcription regulation gene and plays a key role in plant stress response and other stress tolerance processes.In order to investigate the molecular mechanism of heat stress response in garlic,based on the transcriptome data of garlic,the experiment was conducted to obtain AsHSFB1 gene.This gene encoding HSF transcription factor was isolated by homologous cloning of Xusuan NO.6.Sequence analysis indicated that open reading frame(ORF)length of AsHSFB1 was 882 bp,encoding 293 amino acids.Its protein contained characteristic domain of heat shock transcription factor.Phylogenetic tree analysis result showed that AsHSFB1 was close to AT4G11660(AtHSFB2B).Subcellular localization results showed that AsHSFB1 was mainly localized in the nucleus and cytoplasm.In this study,overexpression vector pCAMBIA1305-AsHSFB1 was successfully constructed by homologous recombination technology,which provided a theoretical basis for the subsequent study on the function of AsHSFB1 gene and the cultivation of heat-resistant garlic varieties.RT-PCR analysis showed that the relative expression level of AsHSFB1 gene in garlic leaves was highest,indicating that it had tissue specificity.Compared with the control,the relative expression level of AsHSFB1 gene in Xusuan No.6 increased significantly at 24 h under 38℃high temperature stress.Under 4℃low temperature stress,the relative expression level of AsHSFB1 gene in Xusuan No.6 and Xusuan 815 showed a similar trend,which increased first and then decreased and reached the peak at 4 h.
作者
杨青青
杨峰
赵永强
陆信娟
刘灿玉
葛洁
张碧薇
樊继德
YANG Qing-qing;YANG Feng;ZHAO Yong-qiang;LU Xin-juan;LIU Can-yu;GE Jie;ZHANG Bi-wei;FAN Ji-de(Xuzhou Institute of Agricultural Sciences in Jiangsu Xuhuai Area,Xuzhou 221121,China)
出处
《江苏农业学报》
CSCD
北大核心
2023年第1期169-177,共9页
Jiangsu Journal of Agricultural Sciences
基金
徐州市基础研究计划面上项目(KC21032)
江苏省农业科学院探索性项目[ZX(21)1229]。
关键词
大蒜
AsHSFB1基因
同源重组技术
亚细胞定位
相对表达水平
garlic
AsHSFB1 gene
homologous recombination technology
subcellular localization
relative expression level
