SA、MeJA、ACC和苹果轮纹病病原菌诱导湖北海棠MhWRKY1基因的表达

中国农业科学 ›› 2011, Vol. 44 ›› Issue (5): 990-999 .

SA、MeJA、ACC和苹果轮纹病病原菌诱导湖北海棠MhWRKY1基因的表达

张计育,佟兆国,高志红,罗昌国,渠慎春,章镇

(南京农业大学园艺学院)

收稿日期:2010-08-06 修回日期:2010-12-21 出版日期:2011-03-01 发布日期:2011-03-01
通讯作者: 章镇,渠慎春

Expression of MhWRKY1 Gene Induced by the Elicitors SA,MeJA, ACC and the Apple Ring Spot Pathogen

ZHANG Ji-yu, TONG Zhao-guo, GAO Zhi-hong , LUO Chang-guo,QU Shen-chun, ZHANG Zhen

(南京农业大学园艺学院)

Received:2010-08-06 Revised:2010-12-21 Online:2011-03-01 Published:2011-03-01
Contact: ZHANG Zhen,QU Shen-chun

摘要/Abstract

摘要：

【目的】从湖北海棠叶片中克隆MhWRKY1转录因子的全长cDNA序列,分析该基因在各种组织中(叶、茎、根)的表达特性,并分析SA、MeJA、ACC在叶、茎、根中诱导MhWRKY1基因的表达模式以及苹果轮纹病病原菌诱导条件下湖北海棠叶片中该基因的表达特性。【方法】利用电子克隆技术和RT-PCR验证相结合的方法,从SA诱导的湖北海棠全长cDNA文库中,克隆MhWRKY1转录因子的全长序列;利用生物信息学的方法对其进行结构和功能的初步分析;利用实时荧光定量RT-PCR技术分析该基因在不同组织中的表达以及在SA、MeJA、ACC和苹果轮纹病病原菌诱导下的表达特性。【结果】克隆了MhWRKY1基因的全长cDNA序列为1 338 bp,GenBank数据库登录号为FJ598139。生物信息学分析表明,该基因最大开放阅读框为993 bp,编码330个氨基酸。推导的氨基酸序列与杨树WRKY26、杨树WRKY20、大豆WRKY,马铃薯WRKY2、烟草WRKY、拟南芥WRKY7、水稻WRKY53的同源性分别为68%、68%、66%、60%,59%,49%和43%。该转录因子含有1个WRKY结构域,其N端含有1个WRKYGQK结构域,C端含有1个C2H2锌脂结构,属于第Ⅱ类型的转录因子。表达分析结果表明,MhWRKY1在叶和茎中的表达量较大,分别是根的4.81和3.75倍。在湖北海棠叶、茎、根中,SA、MeJA、ACC都可以诱导该基因的表达。另外,在所研究的72 h内,苹果轮纹病病原菌可以诱导该基因表达,且表达量在12 h达到最大,是未处理之前的9倍左右。【结论】MhWRKY1转录因子可能参与SA、MeJA和ET介导的植物抗病防卫反应的基本信号通路,并且参与对苹果轮纹病病原菌引起的防卫反应,在湖北海棠的的抗病过程中可能起着非常重要的作用。

关键词: 湖北海棠, MhWRKY1, 表达特性, SA, MeJA, ACC, 苹果轮纹病病原菌

Abstract:

【Objective】This study aimed to clone a full length cDNA of MhWRKY1 from Malus hupehensis and analyze the expression pattern of MhWRKY1 in different tissues (leaf, stem and root) and the expression pattern after treatment with the elicitors salicylic acid (SA), methyl jasmonate (MeJA), 1-aminocyclopropane-1-carboxylic acid (ACC) and apple ring spot pathogens (Botryosphaeria berengeriana de Not. f. sp. Parabola Nise Koganezawa et Sakuma), respectively.【Method】The full length cDNA sequence of MhWRKY1 from the M. hupehensis was isolated using the method of in silico cloning combined with the RT-PCR validation. Real-time quantitative RT-PCR (qRT-PCR) was performed to determine the expression pattern of MhWRKY1 in different tissues (leaf, stem and root) and the expression pattern in leaf, stem and root after treatment with SA, MeJA and ACC, respectively. And the expression of this gene after treatment with apple ring spot pathogens was also performed using qRT-PCR.【Result】 1 338 bp full-length cDNA sequence of MhWRKY1 was cloned and the GenBank database accession number was FJ598139. Bioinformatics analysis showed that the open reading frame of this gene was 993 bp, encoding 330 amino acids. The deduced amino acid sequence shares 68%, 68%, 66%, 60%, 59%, 49% and 43% identity with poplar WRKY26, WRKY20, soybean WRKY, potato WRKY2, tobacco WRKY, arabidopsis WRKY7, and rice WRKY53. MhWRKY1 protein contains one conserved WRKY domains that is composed of about 60 amino acid residues with the WRKYGQK sequence followed by a C2H2 zinc finger motif, belongs to the second type of transcription factor. Expression analysis showed that the expression of MhWRKY1 in leaves and stems were high and the expression in leaves and stem were 4.81-fold and 3.75-fold than that in root, respectively. SA, MeJA and ACC induced the expression of MhWRKY1 gene in leave, stem and root, respectively. In addition, the expression of this gene was enhanced and up to the peak at 12 h (9-fold than untreated materials) after treatment with apple ring rots pathogens during the first 72 h.【Conclusion】MhWRKY1 could be involved in SA, MeJA and ET-mediated basic signaling pathway of plant defense responses and disease resistance signal transduction network. MhWRKY1 was involved in defense responses mediated by apple ring rot pathogen. It might be play a key role in disease resistance in M. hupehensis.

Key words: Malus hupehensis (Pamp.) Rehd, MhWRKY1, expression pattern, SA, MeJA, ACC, apple ring rot pathogen (Botryosphaeria berengeriana de Not. f. sp. Parabola Nise Koganezawa et Sakuma)

张计育,佟兆国,高志红,罗昌国,渠慎春,章镇 . SA、MeJA、ACC和苹果轮纹病病原菌诱导湖北海棠MhWRKY1基因的表达
[J]. 中国农业科学, 2011, 44(5): 990-999 .

ZHANG Ji-yu,TONG Zhao-guo,GAO Zhi-hong ,LUO Chang-guo,QU Shen-chun,ZHANG Zhen
. Expression of MhWRKY1 Gene Induced by the Elicitors SA,MeJA, ACC and the Apple Ring Spot Pathogen
[J]. Scientia Agricultura Sinica, 2011, 44(5): 990-999 .

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