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Molecular Cloning and Expression Analysis of Eight PgWRKY Genes in Panax ginseng Responsive to Salt and Hormones

机译:人参对盐和激素反应的八个PgWRKY基因的分子克隆和表达分析

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摘要

Despite the importance of WRKY genes in plant physiological processes, little is known about their roles in Panax ginseng C.A. Meyer. Forty-eight unigenes on this species were previously reported as WRKY transcripts using the next-generation sequencing (NGS) technology. Subsequently, one gene that encodes PgWRKY1 protein belonging to subgroup II-d was cloned and functionally characterized. In this study, eight WRKY genes from the NGS-based transcriptome sequencing dataset designated as PgWRKY2-9 have been cloned and characterized. The genes encoding WRKY proteins were assigned to WRKY Group II (one subgroup II-c, four subgroup II-d, and three subgroup II-e) based on phylogenetic analysis. The cDNAs of the cloned PgWRKYs encode putative proteins ranging from 194 to 358 amino acid residues, each of which includes one WRKYGQK sequence motif and one C2H2-type zinc-finger motif. Quantitative real-time PCR (qRT-PCR) analysis demonstrated that the eight analyzed PgWRKY genes were expressed at different levels in various organs including leaves, roots, adventitious roots, stems, and seeds. Importantly, the transcription responses of these PgWRKYs to methyl jasmonate (MeJA) showed that PgWRKY2, PgWRKY3, PgWRKY4, PgWRKY5, PgWRKY6, and PgWRKY7 were downregulated by MeJA treatment, while PgWRKY8 and PgWRKY9 were upregulated to varying degrees. Moreover, the PgWRKY genes increased or decreased by salicylic acid (SA), abscisic acid (ABA), and NaCl treatments. The results suggest that the PgWRKYs may be multiple stress–inducible genes responding to both salt and hormones.
机译:尽管WRKY基因在植物生理过程中很重要,但对其在人参C.A中的作用知之甚少。迈耶以前使用下一代测序(NGS)技术将该物种上的48个单基因报道为WRKY转录本。随后,克隆了编码属于II-d亚组的PgWRKY1蛋白的一个基因,并对其功能进行了表征。在这项研究中,已经从基于NGS的转录组测序数据集中命名为PgWRKY2-9的8个WRKY基因克隆并鉴定。基于系统发育分析,将编码WRKY蛋白的基因分配给WRKY II组(II-c一个亚组,II-d四个亚组和II-e三个亚组)。克隆的PgWRKY的cDNAs编码了194至358个氨基酸残基的推定蛋白质,每个蛋白质都包含一个WRKYGQK序列基序和一个C2H2型锌指基序。实时定量PCR(qRT-PCR)分析表明,八个被分析的PgWRKY基因在包括叶,根,不定根,茎和种子在内的各种器官中以不同的水平表达。重要的是,这些PgWRKYs对茉莉酸甲酯(MeJA)的转录反应表明,MeJA处理下调了PgWRKY2,PgWRKY3,PgWRKY4,PgWRKY5,PgWRKY6和PgWRKY7,而将PgWRKY8和PgWRKY9上调了度。此外,PgWRKY基因通过水杨酸(SA),脱落酸(ABA)和NaCl处理而增加或减少。结果表明, PgWRKYs 可能是对盐和激素均产生响应的多种压力诱导基因。

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