Nuruzzaman Mohammed, Cao Hongzhe, Xiu Hao, Luo Tiao, Li Jijia, Chen Xianghui, Luo Junli, Luo Zhiyong
Molecular Biology Research Center, State Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China.
Molecular Biology Research Center, State Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China
Acta Biochim Biophys Sin (Shanghai). 2016 Feb;48(2):117-31. doi: 10.1093/abbs/gmv122. Epub 2015 Dec 18.
WRKY proteins belong to a transcription factor (TF) family and play dynamic roles in many plant processes, including plant responses to abiotic and biotic stresses, as well as secondary metabolism. However, no WRKY gene in Panax ginseng C.A. Meyer has been reported to date. In this study, a number of WRKY unigenes from methyl jasmonate (MeJA)-treated adventitious root transcriptome of this species were identified using next-generation sequencing technology. A total of 48 promising WRKY unigenes encoding WRKY proteins were obtained by eliminating wrong and incomplete open reading frame (ORF). Phylogenetic analysis reveals 48 WRKY TFs, including 11 Group I, 36 Group II, and 1 Group III. Moreover, one MeJA-responsive unigene designated as PgWRKY1 was cloned and characterized. It contains an entire ORF of 1077 bp and encodes a polypeptide of 358 amino acid residues. The PgWRKY1 protein contains a single WRKY domain consisting of a conserved amino acid sequence motif WRKYGQK and a C2H2-type zinc-finger motif belonging to WRKY subgroup II-d. Subcellular localization of PgWRKY1-GFP fusion protein in onion and tobacco epidermis cells revealed that PgWRKY1 was exclusively present in the nucleus. Quantitative real-time polymerase chain reaction analysis demonstrated that the expression of PgWRKY1 was relatively higher in roots and lateral roots compared with leaves, stems, and seeds. Importantly, PgWRKY1 expression was significantly induced by salicylic acid, abscisic acid, and NaCl, but downregulated by MeJA treatment. These results suggested that PgWRKY1 might be a multiple stress-inducible gene responding to hormones and salt stresses.
WRKY蛋白属于转录因子(TF)家族,在许多植物生理过程中发挥着重要作用,包括植物对非生物和生物胁迫的响应以及次生代谢。然而,迄今为止,尚未见有关人参WRKY基因的报道。在本研究中,利用新一代测序技术,从该物种茉莉酸甲酯(MeJA)处理的不定根转录组中鉴定出了多个WRKY单基因。通过去除错误和不完整的开放阅读框(ORF),共获得了48个编码WRKY蛋白的有前景的单基因。系统发育分析显示有48个WRKY转录因子,包括11个I组、36个II组和1个III组。此外,克隆并鉴定了一个名为PgWRKY1的MeJA响应单基因。它包含一个1077 bp的完整ORF,编码一个由358个氨基酸残基组成的多肽。PgWRKY1蛋白包含一个由保守氨基酸序列基序WRKYGQK组成的单一WRKY结构域和一个属于WRKY亚组II-d的C2H2型锌指基序。PgWRKY1-GFP融合蛋白在洋葱和烟草表皮细胞中的亚细胞定位显示,PgWRKY1仅存在于细胞核中。实时定量聚合酶链反应分析表明,与叶、茎和种子相比,PgWRKY1在根和侧根中的表达相对较高。重要的是,水杨酸、脱落酸和NaCl显著诱导PgWRKY1的表达,但MeJA处理使其下调。这些结果表明,PgWRKY1可能是一个对激素和盐胁迫有多种应激诱导作用的基因。