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WRKY转录因子作为植物特有的一类重要转录调控因子,广泛参与植物对生物和非生物胁迫的应答反应及形态发育和组织衰老等,已引起人们的关注。本文以前期克隆得到的毛白杨WRKY转录因子基因(命名为PtWRKY1)为研究对象,开展其基因枪介导洋葱表皮细胞瞬时表达、外源信号分子(水杨酸SA,茉莉酸MeJA,脱落酸ABA)诱导表达、转PtWRKY1基因烟草植株的烟草花叶病毒(TMV)接种实验以及抗病相关基因表达的实时荧光定量RT-PCR(qRT-PCR)分析等研究。结果表明,PtWRKY1基因编码蛋白定位于细胞核,SA、ABA及MeJA处理均可诱导PtWRKY1基因表达,但不同诱导因子对PtWRKY1表达量的影响存在一定差异;同时,TMV侵染实验发现外源PtWRKY1表达能增强转基因烟草抗TMV能力,qRT-PCR分析表明PtWRKY1基因及膜保护性酶(POD,SOD,CAT)基因均受TMV诱导而增强表达。本文研究结果可为后续开展毛白杨转录因子PtWRKY1的功能鉴定及应用奠定基础。
WRKY transcription factor, as a kind of plant specific transcriptional regulator, has been widely concerned with the response of plants to biotic and abiotic stress, morphological development and tissue aging. In this paper, the gene of WRKY transcription factor (named as PtWRKY1) was cloned and transiently expressed on onion epidermal cells. The expression of exogenous signal molecules (salicylic acid SA, MeJA, abscisic acid ABA (TMV) inoculation of PtWRKY1 transgenic tobacco plants and real-time quantitative RT-PCR (qRT-PCR) analysis of resistance-related gene expression. The results showed that the PtWRKY1 gene was located in the nucleus. The expression of PtWRKY1 gene was induced by SA, ABA and MeJA, but there were some differences in the expression of PtWRKY1 induced by different inducing factors. Meanwhile, the expression of exogenous PtWRKY1 QRT-PCR analysis showed that PtWRKY1 gene and membrane-protective enzyme (POD, SOD, CAT) genes were all induced by TMV to enhance their expression. The results of this study may lay the foundation for the functional identification and application of Populus tomentosa transcription factor PtWRKY1.