红掌响应细菌性叶疫病的转录因子基因AanWRKY1的鉴定
发布时间：2018-10-05 10:16
【摘要】：WRKY家族转录因子参与植物各种生物胁迫和非生物胁迫反应,同时也参与形态建成、物质代谢、种子休眠、衰老等过程。本研究在前期红掌疫病诱导转录组数据中筛选到一个WRKY家族基因,RT-PCR扩增到1 658 bp全长cDNA,包含一个1 035 bp的开放阅读框,生物信息学分析预测此基因是一个典型的WRKY转录因子,命名为Aan WRKY1(NCBI No.KY597634)。qRT-PCR证实该基因叶片中受疫病侵染上调表达,同时能够被SA、JA、ABA、ETH、GA等激素信号分子诱导,非处理条件下在花梗和茎中表达量最高。通过农杆菌介导洋葱表皮侵染法,瞬时表达AanWRKY1-eGFP,显示该基因编码蛋白定位于细胞核,进一步酵母单杂交实验,证明该基因具有离体转录激活活性。研究结果表明红掌AanWRKY1基因能够响应细菌性疫病及其他胁迫反应,具有应激转录调控的作用,该基因的鉴定为进一步解析其分子病理学作用机理提供依据。
[Abstract]:WRKY family transcription factors are involved in plant stress and abiotic stress responses, but also participate in morphogenesis, substance metabolism, seed dormancy, senescence and other processes. In this study, an WRKY family gene was amplified by RT-PCR from the primordial transcripts of blight. The full-length cDNA, of 1 658 bp contained an open reading frame of 1 035 bp. Bioinformatics analysis predicted that this gene was a typical WRKY transcription factor. Aan WRKY1 (NCBI No.KY597634). QRT-PCR confirmed that the gene was up-regulated by blight infection in leaves and could be induced by hormone signaling molecules such as SA,JA,ABA,ETH,GA, and the highest expression was found in pedicels and stems under untreated conditions. Through Agrobacterium tumefaciens mediated onion epidermal infection, the transient expression of AanWRKY1-eGFP, showed that the gene encoded protein was located in the nucleus. Further yeast single hybrid experiments showed that the gene had in vitro transcriptional activation activity. The results showed that the AanWRKY1 gene could respond to bacterial blight and other stress responses and play a role in stress transcriptional regulation. The identification of the gene provides a basis for further elucidation of its molecular pathological mechanism.
【作者单位】： 海南大学农学院;中国热带农业科学院热带作物品种资源研究所;
【基金】：国家自然科学基金青年项目(31201652);国家自然科学基金面上项目(31471902)共同资助
【分类号】：S436.8
，
本文编号：2253005
[Abstract]:WRKY family transcription factors are involved in plant stress and abiotic stress responses, but also participate in morphogenesis, substance metabolism, seed dormancy, senescence and other processes. In this study, an WRKY family gene was amplified by RT-PCR from the primordial transcripts of blight. The full-length cDNA, of 1 658 bp contained an open reading frame of 1 035 bp. Bioinformatics analysis predicted that this gene was a typical WRKY transcription factor. Aan WRKY1 (NCBI No.KY597634). QRT-PCR confirmed that the gene was up-regulated by blight infection in leaves and could be induced by hormone signaling molecules such as SA,JA,ABA,ETH,GA, and the highest expression was found in pedicels and stems under untreated conditions. Through Agrobacterium tumefaciens mediated onion epidermal infection, the transient expression of AanWRKY1-eGFP, showed that the gene encoded protein was located in the nucleus. Further yeast single hybrid experiments showed that the gene had in vitro transcriptional activation activity. The results showed that the AanWRKY1 gene could respond to bacterial blight and other stress responses and play a role in stress transcriptional regulation. The identification of the gene provides a basis for further elucidation of its molecular pathological mechanism.
【作者单位】： 海南大学农学院;中国热带农业科学院热带作物品种资源研究所;
【基金】：国家自然科学基金青年项目(31201652);国家自然科学基金面上项目(31471902)共同资助
【分类号】：S436.8
，
本文编号：2253005
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