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水稻WRKY80转录调节蛋白基因的分离与表达模式
  • ISSN号:0578-1752
  • 期刊名称:中国农业科学
  • 时间:2013.10.1
  • 页码:4035-4043
  • 分类:S511[农业科学—作物学]
  • 作者机构:[1]湖南科技大学生命科学学院,湖南湘潭411201
  • 相关基金:国家自然科学基金(31301617,31171803)、国家公益性(农业)行业科研专项(nyhyzx3-16)、湖南省自然科学基金重点项目(10JJ2030)
  • 相关项目:OsWRKY4转录因子在水稻对纹枯病阶段抗性中的调节作用及其分子机理研究
中文摘要:

【目的】分离水稻WRKY80,分析其编码蛋白的序列结构特征,了解其在各器官中以及病原接种、激素处理下的表达模式,为阐明其生物学功能奠定基础。【方法】基于水稻基因组数据库注释的Loc_Os03g63810基因的编码序列设计特异引物,用RT-PCR法从茉莉酸甲酯(MeJA)处理的水稻叶片中克隆WRKY80 cDNA 序列,运用生物信息学手段对推定的蛋白和启动子序列进行分析,用Northern杂交或实时荧光定量PCR方法研究基因的表达模式。【结果】 获得了WRKY80 cDNA序列,长1 392 bp,包含1个长1 164 bp 的完整开放读码框,编码387个氨基酸。WRKY80具有1个典型的WRKY 保守结构域,锌指类型为C2H2,归类于WRKY 第Ⅱ组;C-端为酸性结构区,且含连续的6个谷氨酰胺和8个丝氨酸,提示具有转录激活活性;预测其定位于细胞核。WRKY80与玉米和高粱等单子叶植物WRKY 的氨基酸序列同一性较高。WRKY80在各器官中组成性表达,在叶、根和穗中表达丰度较高,花中次之,在茎和颖果中丰度较低,且表达具有发育阶段相关性,在成熟叶、根中的表达分别高于幼叶和幼根;受稻瘟病菌、纹枯病菌、MeJA和乙烯利诱导表达,而水杨酸对其表达无影响。其表达模式与启动子顺式元件预测分析的结果基本一致。【结论】WRKY80具有转录因子的结构特征,推测其可能通过茉莉酸/乙烯介导的信号途径参与水稻的防御反应,也可能参与发育调控。

英文摘要:

【Objective】 The objective of this study is to isolate rice WRKY80, to analyze the characteristics of its coding sequence and to investigate its expression patterns in different organs and under pathogen inoculation and hormone application, thus providing a basis for its function identification. 【Method】Specific primer sequences were designed according to the annotated gene Loc_Os03g63810 released in rice genome database. RT-PCR was used to amplify WRKY80 cDNA sequence from RNA pools generated from methyl jasmonate (MeJA)-treated rice leaves. Bioinformatical tools were employed to analyze its deduced protein sequence and cis-elements in its promoter. Northern blot or real-time fluorescence quantification PCR was used to investigate its expression patterns. 【Result】 The obtained cDNA sequence of WRKY80 was 1 392 bp in length, containing an entire open reading frame of 1 164 bp, encoding a polypeptide of 387 amino acid residues consisting of one classic conserved WRKY domain with a zinc finger motif of C2H2, belonging to the WRKY subgroup Ⅱ. WRKY80 possessed an acidic C terminus with consecutive 6 glutamines and 8 threonines, an acidic region possibly responsible for transcription activating activity, and was predicted to be localized in nucleus. WRKY80 shared high identity at the amino acid level with those from monocotyledons Zea mays and Sorghum bicolor. WRKY80 was constitutively expressed in all tested organs. The transcript abundance was relatively higher in leaves, roots and panicles, next was in flowers, and less was in stems and grains. WRKY80 expression was higher in mature leaves and roots than in young leaves and roots, respectively, indicating a developmental stage-related feature. It was rapidly induced by inoculation with Magnaporthe oryzae and Rhizoctonia solani, and also by application of exogenous MeJA and ethephon, whereas salicylic acid exerted no effects on its expression. Consistent with the expression profiles was the prediction result of cis-elements in its promoter. 【Concl

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期刊信息
  • 《中国农业科学》
  • 中国科技核心期刊
  • 主管单位:中华人民共和国农业部
  • 主办单位:中国农业科学院 中国农学会
  • 主编:万建民
  • 地址:北京中关村南大街12号中国农业科学院图书馆楼4101-4103室
  • 邮编:100081
  • 邮箱:zgnykx@caas.cn
  • 电话:010-82109808 82106279
  • 国际标准刊号:ISSN:0578-1752
  • 国内统一刊号:ISSN:11-1328/S
  • 邮发代号:2-138
  • 获奖情况:
  • 中国期刊方阵“双高”期刊,第三届中国出版政府奖提名奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),英国农业与生物科学研究中心文摘,波兰哥白尼索引,英国动物学记录,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),英国食品科技文摘,中国北大核心期刊(2000版)
  • 被引量:85620