樱桃砧木PcWRKY1基因的克隆与表达分析

摘要

WRKY转录调控因子广泛参与植物的生长发育、生物胁迫和非生物胁迫等生理过程.为明确干旱和盐等非生物胁迫对樱桃砧木PcWRKY1基因表达的影响,本研究利用RT-PCR技术克隆了樱桃砧木Gisela 6(Prunus cerasus × Prunus canescens)的WRKY基因cDNA序列,命名为PcWRKY1(GenBank登录号为KY399985).序列分析结果表明,PcWRKY1完整开放阅读框为1 461 bp,编码486个氨基酸,编码的蛋白质含有2个WRKY保守结构域和2个C2H2锌指结构域,属于WRKY家族第Ⅰ类成员. qRT-PCR分析PcWRKY1在200 mmol甘露醇和200 mmol NaCl胁迫下的表达情况,结果显示,甘露醇和NaCl均能诱导PcWRKY1基因表达,推测PcWRKY1基因在应对干旱和盐2种非生物胁迫方面可能具有重要作用.%The WRKY transcription factors are widely involved in the physiological processes related to plant growth and development, biotic and abiotic stresses. A full-length cDNA for PcWRKY1 (GenBank accession number KY399985) gene was cloned from the cherry (Prunus) rootstock Gisela 6(Prunus cerasus×Prunus canescens) using reverse transcrip-tion-polymerase chain reaction (RT-PCR). Sequence analysis results showed that the PcWRKY1 gene was found to contain an open reading frame (ORF) with 1 461 bp and encoded 486 amino acids. The protein encoded by PcWRKY1 comprised of two conserved WRKY domains and two zinc finger structures (C2H2), and thus, belonged to Group I of the WRKY family. Furthermore, the expression of PcWRKY1 under the stresses of 200 mmol mannitol and 200 mmol NaCl was analysed by quantitative RT-PCR. Both mannitol and NaCl could induce the expression of PcWRKY1 gene, suggesting that PcWRKY1 played an important role in dealing with drought and salt stress.