棉花AP2/ERF-B3亚组转录因子基因GhERF14的克隆与表达分析

Cloning and Expression Analysis of a Transcription Factor Gene GhERF14 from Cotton AP2/ERF-B3 Subgroup

为了探究GhERF14基因在抗棉花枯萎病中的调控作用,利用RT-PCR技术从陆地棉‘中棉所12’中克隆一个新的AP2/ERF转录因子基因GhERF14,并对其表达量和生物信息学进行了分析。结果表明,GhERF14无内含子,开放阅读框(ORF)为414 bp,编码137个氨基酸,分子式为C_(663)H_(1027)N_(199)O_(216)S_(4)。亚细胞定位预测编码的Gh ERF14蛋白主要分布在细胞核中。GhERF14蛋白含有3个信号肽,无跨膜结构,属于亲水蛋白;含有1个保守的AP2结构域,属于B3亚组;预测GhERF14基因启动子区域有乙烯(ET)响应元件、脱落酸(ABA)响应元件、伤口响应以及生物胁迫响应元件等。组织特异性分析显示该基因在茎部表达量最高,其次是根,最后是叶和茎尖。枯萎病菌(Fusarium oxysporum)和激素处理发现,该基因受枯萎病菌诱导后,呈先增加后降低再升高的动态变化趋势,并且在6 h达到最高。ET、JA、SA三种激素诱导后,该基因相对表达量均呈上调表达趋势,推测该基因可能通过乙烯(ET)、茉莉酸(JA)、水杨酸(SA)信号通路参与棉花对枯萎病的胁迫应答,以上结果为后续进一步研究该基因功能奠定基础。

To investigate the regulatory role of the GhERF14 gene in resistance to cotton wilt,a new AP2/ERF transcription factor gene GhERF14 was cloned from’Zhongmiansuo12’in upland cotton by RT-PCR,and its expression and bioinformatics were analyzed.The results showed that GhERF14 had no introns,the open reading frame(ORF)was 414 bp,coding 137 amino acids.The molecular formula was C_(663)H_(1027)N_(199)O_(216)S_(4).The GhERF14 protein encoded by subcellular localization prediction was mainly distributed in the nucleus.GhERF14 protein contained three signal peptides,had no transmembrane structure,and belonged to hydrophilic protein.It contained a conserved AP2 domain and belonged to B3 subgroup.The promoter region of GhERF14 gene was predicted to contain ethylene(ET)response element,abscisic acid(ABA)response element,wound response element and biological stress response element.Tissue specificity analysis showed that the gene expression was highest in roots,followed by leaves and stems.Fusarium oxysporum and hormone treatment showed that,after being induced by Fusarium oxysporum,the gene showed a dynamic change trend of first increasing,then decreasing and then increasing,and reached its peak at 6 h.After the induction of ET,JA and SA,the relative expression of this gene was up-regulated.It is speculated that this gene may be involved in the stress response of cotton to wilt through ethylene(ET),jasmonic acid(JA)and salicylic acid(SA)signaling pathways.These results lay a foundation for further study on the function of this gene.