富士苹果MdWRKY40b基因克隆及其对白粉病的抗性分析
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贵州省农业科学院研究生科研创新基金


Cloning of MdWRKY40b Gene in Fuji Apple and Its Response to Powdery Mildew Stress
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    摘要:

    WRKY是植物抗病相关的一个大转录因子家族基因,为了揭示WRKY40基因在调控白粉病抗性方面的作用,以富士苹果(Malus domestica Borkh.cv.Fuji)为试材,利用特异引物进行RT-PCR基因克隆,并用实时荧光定量PCR方法分析组织中基因的表达及接种苹果白粉菌和用1 mmol·L-1水杨酸(SA)喷施处理后对基因表达的影响。结果表明:(1)克隆到的片段长度为1 240 bp,包含一个完整的开放阅读框,长1 002 bp,编码334个氨基酸,其氨基酸序列与AtWRKY40、HvWRKY1/2/3等8个序列一致性为48.96%;该基因与拟南芥AtWRKY40同源,故命名为MdWRKY40b,GenBank登录号为JX112907。(2)基因表达分析表明,MdWRKY40b在苹果根、茎、叶、花均有表达,其中根、叶、花中表达量基本相当,茎中表达量最低。(3)该基因明显受苹果白粉菌和SA的强烈诱导表达且持续时间较长,并且两者表达趋势相同、出现最高表达量的时间相近。研究推测,富士苹果容易感白粉病的主要原因可能与MdWRKY40b的持续长时间的高表达有关。

    Abstract:

    WRKY is the genes of a super transcription factors family that related to disease resistance.In order to reveal the role of WRKY40 gene in response to powdery mildew stress in apple.Malus domestica Borkh.cv.Fuji was used for gene cloning by reverse transcription PCR amplification by using specific primers.Quantitative real-time PCR was used in analyzing gene expression in tissues and in response to Podosphaera leucotricha and treatment of 1 mmol·L-1 SA(salicylic acid).(1)A 1 240 bp fragment including a 1 002 bp open reading frame to encode 334 amino acids was isolated.The amino acids sequence share 48.9% identity BLASTp to AtWRKY40,HvWRKY1/2/3 and others 4 sequences.It is a homologous gene of Arabidopsis AtWRKY40,and it is named as MdWRKY40b.Its GenBank accession is JX112907.(2)Gene expression indicated that MdWRKY40b expressed in root,stem,leaf and flower,with almost equally expression level in leaf,root and flower but very low level in stem.(3)It was clearly induced by P.leucotricha,and exogenous SA for a long times,with similar expression trends and the highest expression level time.The reason of Fuji apple extremely susceptible to powdery mildew maybe relate to the long-time and high expression of MdWRKY40b.

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罗昌国,袁启凤,裴晓红,等.富士苹果MdWRKY40b基因克隆及其对白粉病的抗性分析[J].西北植物学报,2013,33(12):2382-2387

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  • 在线发布日期: 2013-12-30
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