红掌响应细菌性叶疫病的转录因子基因AanWRKY1的鉴定

Identification of Transcription Factor Gene AanWRKY1 Response to Bacterial Leaf Blight in Anthurium andraeanum

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。

WRKY family transcription factors not only participate in various plant biological stress and abiotic stress response, but also involve in morphogenesis, material metabolism, seed dormancy, aging and other processes. In this research, a WRKY family gene was screened from the data of the previous transcription group, and RT-PCR was amplified to the full length eDNA of 1 658 bp, which contained a 1 035 bp open reading frame. Bioinformatics analysis predicted that this gene was a typical WRKY transcription factor, which named A anWRKY1 （NCBI No.KY597634）. qRT-PCR confirmed that the gene expression was up-regulated in the leaves, and could be induced by hormone signal molecules such as SA, JA, ABA, ETH and GA, and the highest expression in pedicels and stems under untreated conditions. The AanWRKYl-eGFP was instantaneous expressed by Agrobacterium tumefaciens on the onion epidermis, which indicated that the gene was located in the nucleus, and further yeast one-hybrid experiment proved that the gene had inactivated transcriptional activity. The results showed that the AanWRKY1 gene was able to respond to bacterial epidemics and other stress response, it had the effect of stress transcription regulation. The identification of this gene laid the foundation for further analysis of its molecular pathology mechanism.