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WRKY转录因子是高等植物中最重要的转录因子基因家族之一,其在植物形态建成、生长发育以及应对生物(病菌、害虫等)、非生物(干旱、盐、冷、热等)胁迫过程中发挥重要作用。该研究根据人参化感自毒物质苯甲酸诱导的人参转录组数据,设计全长扩增引物,利用RT-PCR方法获得1条编码WRKY转录因子的基因,暂命名为WRKY7,并对其进行TA克隆、测序和序列分析。该基因cDNA全长1 216 bp,其开放阅读框(open reading frame,ORF)为1 014 bp,编码337个氨基酸。序列分析及系统进化分析表明,该蛋白属于WRKY家族第Ⅲ类成员,与西洋参种的WRKY6转录因子亲缘关系最近,同源性为87%。实时荧光定量PCR分析发现,苯甲酸胁迫后WRKY7的表达量显著升高,推测该基因可能参与了人参响应苯甲酸胁迫的过程,为进一步研究人参自毒胁迫响应的分子机制奠定了基础。
WRKY transcription factor is one of the most important gene family of transcription factors in higher plants. It is widely used in plant morphogenesis, growth and development as well as in dealing with the biological (germs, pests etc), abiotic (drought, salt, cold and heat) Play an important role. In this study, a full-length amplification primer was designed based on the ginseng transcriptome data induced by benzoic acid in the ginseng. A gene encoding WRKY transcription factor was obtained by RT-PCR and named as WRKY7 Cloning, sequencing and sequence analysis. The full-length cDNA of this gene is 1 216 bp in length and has an open reading frame (ORF) of 1 014 bp encoding 337 amino acids. Sequence analysis and phylogenetic analysis indicated that the protein belonged to the third member of WRKY family, which had the closest genetic relationship with WRKY6 transcription factor of American ginseng, with a homology of 87%. Real-time PCR analysis showed that the expression of WRKY7 was significantly increased after benzoic acid stress, suggesting that the gene may be involved in the process of benzoic acid stress response to ginseng, which laid the foundation for further study on the molecular mechanism of ginseng autotoxic stress response.