六种重要瓜类种传细菌可视基因芯片筛查方法研究  

Six Seedborne Bacterial Pathogens Screening with Chromogenic DNA Macroarray in Melon Crops

作　　者：张靓 田茜[1] 刘凤权[2] 朱水芳[1] 赵文军[1] 

机构地区：[1]中国检验检疫科学研究院,北京100029 [2]南京农业大学植物保护学院,南京210095

出　　处：《农业生物技术学报》2013年第1期120-126,共7页Journal of Agricultural Biotechnology

基　　金：国家"十二五"科技支撑计划(No.2012BAK11B02;No.2012BAK11B06);质检总局科技计划项目(No.2011IK185)

摘　　要：种传细菌是瓜类作物及健康种子生产中关注的重要对象。针对细菌性白枯病菌(Pseudomonas viridiflava)、细菌性角斑病菌(P.syringae pv.lachrymans)、丁香假单胞菌丁香致病变种(P.syringae pv.syringae)、瓜类果斑病菌(Acidovorax citrulli)、细菌性叶斑病菌(Xanthomonas cucurbitae)、甜瓜黄单胞菌(X.melonis)6种重要瓜类种传细菌,利用多重PCR扩增及后续可视芯片检测技术,建立了可视基因芯片筛查方法。实验结果表明,以细菌纯培养液为模板,6种目标菌检测灵敏度平均可达N×103～N×104CFU/mL,并能从人工模拟带菌的西瓜(Citrullus vulgaris)、甜瓜(Euphorbia meloformis)、黄瓜(Cucumis sativus)的种子样品及自然感染瓜类果斑病菌的西瓜种子中检测到目标细菌。该研究为瓜类种传细菌的快速初筛及健康种子质量控制提供了新手段。Seedbome bacteria, including Pseudomonas viridiflava （Pv）, P. syringae pv. lachrymans （Psi）, P. syringae pv. syringae （Pss）, Acidovorax citrulli （Ac）, Xanthomonas cucurbitae （Xc） and X. melonis （Xm）, are important diseases in melon crops production. By using three sets of multiplex PCR and a chromogenic macroarray system, it was allowed us to detect multiple target bacterial species in bacterial suspension, naturally infected or artificially infected seed samples simultaneously without the use of agarose gel electrophoresis. The average qualitative detection limit could be reach N×10^3-N×10^4CFU/mL with pure bacteria culture. Target bacteria could be detected from artificial infected watermelon（Citrullus vulgaris）, melon （Euphorbia meloformis） and cucumber （Cucumis sativus） seeds and naturally infected A. citrulli watermelon seeds. The new method is efficient, high-throughput and practical, and gets rid of the dependence of fluorescence scanner. This study provides a method for the simultaneous detection of plant seedbome pathogens.

关 键 词：瓜类 种传细菌 多重PCR 可视基因芯片 检测方法 

分 类 号：S642.034[农业科学—蔬菜学]