芝麻CCCH锌指蛋白基因SiC3H1的克隆及表达分析

Cloning and Expression Analysis of CCCH Zinc Finger Protein Gene SiC3H1 from Sesame

CCCH锌指蛋白是真核生物中广泛存在的一类转录调节因子,在植物生长发育及逆境胁迫应答中具有重要功能。本研究基于芝麻转录组数据,筛选并克隆到一个芝麻干旱应答相关的CCCH锌指蛋白基因SiC3H1,并对其在不同非生物逆境胁迫下的表达变化进行了研究。测序结果表明,SiC3H1基因编码序列长1 110 bp,编码369个氨基酸。对其编码蛋白进行序列分析发现,该蛋白含有2个CCCH型锌指结构域。进化树分析表明,SiC3H1与番茄和马铃薯中的同源基因相似度较高。启动子序列分析显示,SiC3H1启动子区域含有参与防御和胁迫应答的元件、脱落酸应答元件和乙烯应答元件。实时荧光定量PCR分析表明,在渗透、高盐、低温、高温和淹水5种逆境胁迫处理下,SiC3H1的表达量与对照相比均显著升高。以上研究结果表明,SiC3H1参与了芝麻对多种逆境胁迫的应答反应,为今后进一步解析其功能提供了理论依据。

CCCH zinc finger proteins, a group of transcription regulatory factors widely existed in eukaryotes, play important roles in plant growth, development and stress response. In this study, a drought-responsive CCCH zinc finger gene, SiC3H1 was isolated from sesame, and then the expression of SiC3H1 under various abiotic stress treatments was investigated. The sequencing results indicated that the full length of SiC3H1 coding sequence is 1 110 bp long, encoding 369 amino acids. SiC3H1 protein contained two CCCH zinc finger domains. Based on phylogenetic tree constructed, SiC3H1 was highly similar to the homologous genes in tomato and potato. Promoter region of SiC3H1 contains abscisic acid-responsive element, ethylene-responsive element and cis-element involved in defense and stress responsiveness. Real-time quantitative PCR analysis showed that the expressions of SiC3H1 were significantly upregulated under various stress treatments, including osmosis, high salt, low salt, high temperature,waterlogging. These results suggest that SiC3H1 may be involved in the response of sesame to various stresses, providing a theoretical basis for further analyzing its functions in the future.