葡萄乙醛脱氢酶VvALDH10A9基因的克

发布时间：2018-06-20 05:00

本文选题：葡萄 + 葡萄溃疡病　；参考：《植物保护》2017年05期

【摘要】：由葡萄座腔菌科Botryosphaeriaceae真菌引起的葡萄溃疡病Botryosphaeria dieback是葡萄上的主要枝干病害,严重影响葡萄产量和品质。鉴定和分析参与葡萄与葡萄溃疡病菌互作的基因,有助于揭示和阐明葡萄抗溃疡病的信号通路。本研究根据葡萄溃疡病菌侵染后的葡萄转录组数据信息,利用逆转录聚合酶链式反应技术(RTPCR)克隆了一个受葡萄溃疡病菌诱导上调表达的乙醛脱氢酶基因VvALDH10A9(Vitis vinifera aldehyde dehydrogenase 10A9)。系统进化树分析表明,该基因编码的蛋白与拟南芥AtALDH10A8亲缘关系最近。利用实时荧光定量PCR分析VvALDH10A9的表达结果表明:VvALDH10A9的表达具有组织特异性,在茎、叶和花中表达量最高,在根中表达量相对较低;葡萄溃疡病菌侵染后,VvALDH10A9在葡萄抗病品种中表达变化不明显,而在葡萄感病品种中该基因显著上调表达。利用原核蛋白表达系统诱导合成的VvALDH10A9蛋白,经纯化后可降解乙醛,表明VvALDH10A9蛋白具有乙醛脱氢酶活性。
[Abstract]:Botryosphaeria dieback, which is caused by Botryosphaeriaceae fungi, is the main branch and stem disease of grape, which seriously affects the yield and quality of grape. Identification and analysis of the genes involved in the interaction of grape ulcer pathogen may help to reveal and clarify the signaling pathway of grape resistance to ulcerative disease. In this study, based on the transcriptional data of grape infected by Grape ulcer pathogen, a gene VvALDH10A9Vitis vinifera aldehyde dehydrogenase 10A9 was cloned by reverse transcriptase polymerase chain reaction (RT-PCR). Phylogenetic tree analysis showed that the protein encoded by the gene was closely related to Arabidopsis thaliana AtALDH10A8. The expression of VvALDH10A9 was analyzed by real-time fluorescence quantitative PCR. The results showed that the expression of VvALDH10A9 was tissue specific, the highest in stem, leaf and flower, and the lower in root. The expression of VvALDH10A9 was not significantly changed in resistant grape varieties after infection, but it was upregulated significantly in susceptible grape varieties. VvALDH10A9 protein was induced by prokaryotic expression system and degraded to acetaldehyde after purification, indicating that VvALDH10A9 protein has acetaldehyde dehydrogenase activity.
【作者单位】：北京市农林科学院植物保护环境保护研究所;
【基金】：国家葡萄产业技术体系(CARS-30) 北京市博士后工作经费北京市农林科学院博士后基金
【分类号】：S436.631.1

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