克隆并鉴定冬枣转录因子ZjERF098明确其分子特性,验证ZjERF098瞬时过表达对枣果实黑斑病的抑制效果,为枣果实抗病机制研究奠定基础。对ZjERF098进行系统发育树构建、同源序列比对、结构域分析、蛋白质结构预测、启动子顺式作用元件分析,克隆并构建了pEAQ-ZjERF098等多个重组载体,在烟草上进行亚细胞定位和转录激活活性分析,在冬枣果实上进行瞬时过表达。结果显示,ZjERF098的开放阅读框包含426个碱基对,编码141个氨基酸,与拟南芥中的AtERF098序列相似度最高,同属于ERF转录因子亚家族,只有1个AP2结构域,不含信号肽与跨膜结构域。启动子区域含有与植物防御反应相关的脱落酸响应元件、茉莉酸响应元件等响应元件。ZjERF098具有转录激活活性,主要定位在细胞核上,同时在细胞质膜也有GFP荧光信号。同时,ZjERF098瞬时过表达能够增强枣果实对黑斑病的抗病性,显著降低枣果实的发病率与病斑直径,表明ZjERF098是在枣果实对黑斑病抗病性中起到重要作用的转录因子。
ZjERF098 was cloned and characterized to clarify its molecular properties and to verify the inhibitory effect of transient overexpression of ZjERF098 on black spot rot of jujube, so as to lay the foundation for the research on the mechanism of resistance to disease of jujube fruit.The phylogenetic tree construction, homologous sequence comparison, structural domain analysis, protein structure prediction and promoter cis-acting element analysis were performed on ZjERF098.Several recombinant vectors such as pEAQ-ZjERF098 were constructed to analyze the subcellular localization and activation of transcriptional activity of ZjERF098 in tobacco, and its transient overexpression effect in jujube fruit.The results showed that the open reading frame of ZjERF098 comprises 426 base pairs (bp), encoding 141 amino acids.It shared the highest sequence similarity with the AtERF098 sequence in Arabidopsis thaliana, belonging to the ERF transcription factor subfamily.ZjERF098 had only one AP2 domain and lacked a signal peptide and transmembrane domains.The promoter region contained response elements related to plant defense responses, such as abscisic acid responsive elements and jasmonic acid responsive elements.ZjERF098 exhibited transcriptional activation activity, primarily localizing in the cell nucleus, with GFP fluorescence signals also observed on the cytoplasmic membrane.Simultaneously, transient overexpression of ZjERF098 enhanced the resistance of jujube fruits to black spot rot, significantly reducing disease incidence and lesion diameter.These indicated that ZjERF098 play a crucial role as a transcription factor in enhancing the resistance of jujube fruits to black spot rot.
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