胡萝卜ABF转录因子基因DcABF1的克隆与非生物胁迫应答分析
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引用本文:郝建楠,王雨薇,段奥其,冯 凯,却 枫,熊爱生.胡萝卜ABF转录因子基因DcABF1的克隆与非生物胁迫应答分析[J].西北植物学报,2019,39(10):1731~1740
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郝建楠,王雨薇,段奥其,冯 凯,却 枫,熊爱生* (南京农业大学 园艺学院作物遗传与种质创新国家重点实验室农业农村部华东地区园艺作物生物学与种质创制重点实验室南京 210095) 
基金项目:国家自然科学基金(31872098);
中文摘要:ABF转录因子是一种碱性亮氨酸拉链类蛋白,与植物的生长发育和逆境调控密切相关。该研究以胡萝卜‘君川红’为材料,克隆出编码ABF转录因子的基因DcABF1,分析其编码氨基酸的序列组成、进化关系和蛋白质理化性质等;采用实时荧光定量PCR方法检测DcABF1基因在不同非生物胁迫下的应答模式。结果显示:DcABF1基因含有一个长1 293 bp开放阅读框,编码430个氨基酸,其蛋白质分子量为104.96 kD,理论等电点为4.85。对DcABF1蛋白与其他植物的ABF家族成员进行同源性比对与进化树分析表明,DcABF1与榴莲中的ABF亲缘关系最近。蛋白质功能域预测发现,DcABF1蛋白中有28.84%的α螺旋和56.05%的无规则卷曲。DcABF1基因启动子区域含有多个顺式调控元件。实时荧光定量PCR分析表明,DcABF1基因对高温、低温、盐胁迫均有响应,其表达水平在盐处理下均高于对照,且于处理后4 h达到峰值,呈先上升后下降的趋势,但干旱处理下DcABF1基因的表达水平与对照相比均下降。
中文关键词:胡萝卜  ABF  转录因子  启动子分析  非生物胁迫  基因表达
 
Cloning and Response Analysis to Abiotic Stress of ABF Transcription Factor Gene DcABF1 in Carrot
Abstract:ABF transcription factor (TF) is a basic leucine zipper protein, which involved in the growth, development and stress regulation in plants. In this study, a DcABF1 gene encoding ABF transcription factor was cloned from the carrot cv. ‘Junchuanhong’. The amino acid sequences, phylogenetic evolutionary relationships and physicochemical properties of DcABF1 were analyzed. The relative expression levels of the DcABF1 gene under different abiotic stress treatments were detected by quantitative real time PCR. Sequence analysis indicated that the length of DcABF1 gene was 1293 bp and encoded 430 amino acids. The relative molecular mass of DcABF1 and theoretical pI were 104.96 kD and 4.850, respectively. The sequence alignment and phylogenetic analysis of the DcABF1 with the ABF TFs from other plants showed that the DcABF1 had the closest relationship with ABF from durian. The prediction of protein functional domain revealed that DcABF1 protein contained 28.84% α helix and 56.05% random coil. There were some cis elements in the region of DcABF1 promoter. Quantitative real time PCR analysis indicated that the DcABF1 gene had response to high temperature, low temperature, and salt stress treatments. The expression levels of DcABF1 were up regulated under salt treatment and peaked at 4 h. The expression level trend of DcABF1 under high salt treatment was first increased and then decreased. However, the expression of DcABF1 gene was down regulated compared with the control under drought treatment.
keywords:carrot  ABF  transcription factors  analysis of promoter  abiotic stress  gene expression
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