2008-CMV侵染胁迫下黄瓜重要功能基因表达及代谢响应的研究

中国农业科学 2008,41(11):3691-3697

Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2008.11.033 CMV侵染胁迫下黄瓜重要功能基因表达及代谢响应的研究

毛伟华1,2,龚亚明3,宋兴舜1,夏晓剑1,王彦杰1,周艳虹1,师 恺1,李亚丹1,4,喻景权1

(1浙江大学园艺系/农业部园艺植物生长发育与品质调控重点开放实验室,杭州 310029;2浙江大学分析测试中心,杭州310029;

3浙江省农业科学院蔬菜研究所,杭州 310021;4河北师范大学生命科学学院,石家庄 050016)

摘要:【目的】研究黄瓜花叶病毒病(CMV)侵染胁迫下黄瓜重要功能基因表达和代谢途径响应。【方法】在结合气体交换和叶绿素荧光参数及抗性相关酶活性测定的基础上, 应用农业部园艺植物生长发育与品质调控重

点开放实验室自主开发的黄瓜cDNA芯片研究黄瓜在CMV侵染胁迫下的基因表达谱变化,并对其中5个具有代表性

的基因通过实时荧光定量PCR(QRT—PCR)进行检测,以验证cDNA芯片杂交结果的可靠性。【结果】共获得67

个差异表达显著的基因,其中42个基因下调表达,25个基因上调表达。这些差异表达的基因涉及了许多重要代

谢途径。许多与光合作用、氮同化相关的基因受到明显的抑制;而一些防御相关基因和信号传导相关基因则诱导

表达;同时,也发现了一些与CMV胁迫相关的功能未知基因或未有任何功能信息的基因。【结论】CMV胁迫引发

了黄瓜代谢发生一系列复杂的适应性变化,PR1-1a等基因可能在黄瓜防御CMV侵染过程发挥着重要作用。

关键词:黄瓜;CMV;cDNA芯片;基因表达谱

Expression Analysis of Important Functional Genes and Pathways in Cucumber Leaves in Response to CMV Stress

MAO Wei-hua1,2, GONG Ya-ming3, SONG Xing-shun1, XIA Xiao-jian1, WANG Yan-jie1, ZHOU Yan-hong1,

SHI Kai1, LI Ya-dan1,4, YU Jing-quan1

(1Department of Horticulture, Zhejiang University/The Ministry of Agricultural Laboratory of Horticultural Plant Growth Development and Biotechnology, Hangzhou 310029; 2Center of Analysis and Measurement , Zhejiang University, Hangzhou 310029; 3Institute of Horticulture, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021; 4College of Life

Science, Hebei Normal University, Shijiazhuang 050016)

Abstract: 【Objective】The aim of this paper is to study on the expression of important functional genes and pathways in cucumber leaves in response to CMV stress.【Method】On the basis of study of photosynthesis and activities of defence-related enzymes, the changes of gene expression profiles was analyzed by microarray in cucumber (Cucumis sativus L.) after CMV infection.

In order to validate the microarray result, five representative clones induced or repressed from microarray analysis were selected for

real-time quantitative PCR analysis. 【Result】Significant changes in transcript abundance were observed for 67 genes, among them,

42 showed down-regulated expression and 25 showed up-regulated expression. Analysis of these CMV-responsive genes showed that

they belong to wide range of functional categories. Many genes for photosynthesis and nitrogen assimilation were strongly repressed

while some defense-related genes, on the other hand, were found to be up-regulated. In addition to these expression changes, some functions unknown or unreported novel genes that respond to CMV stress were also identified.【Conclusion】A complex set of metabolic adaptation appears to occur during CMV stress and some genes, such as PR1-1a might play an important role in defence.

Key words: Cucumis sativus L.; CMV; Microarray ; Gene expression profiling

收稿日期:2007-08-25;接受日期:2007-10-15

基金项目:国家重点基础研究发展计划资助项目(2009CB119000),国家自然科学基金资助项目(30400297,30671257,30800763,3050044)和浙江省自然科学基金资助项目(Y307168)

作者简介:毛伟华(1970-),女,浙江温岭人,博士,研究方向为蔬菜分子生物学。Tel:0571-********; E-mail:whmao@http://m.wendangku.net/doc/212cdd94284ac850ac024224.html 。通讯作者喻景权(1963-),男,浙江义乌人,教授,研究方向为设施园艺学。E-mail:jqyu @ zju. http://m.wendangku.net/doc/212cdd94284ac850ac024224.html

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