基质辅助激光解吸/电离飞行时间质谱法鉴定检疫性病菌炭疽菌  

Identification of quarantine colletotrichum isolates by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

作　　者：程颖慧 王颖[2] 杨晓朱 章桂明[2] 冯建军[2] 汪莹[2] CHENG Ying-hui;WANG Ying;YANG Xiao-zhu;ZHANG Gui-ming;FENG Jian-jun;WANG Ying(Shenzhen Fairy Lack Botanical Garden,Shenzhen 518000,China;Shenzhen Entry-Exit Inspection and Quarantine Bureau,Shenzhen 518045,China;Guangdong Eco-engineering Polytechnic,Guangzhou 510520,China)

机构地区：[1]深圳市仙湖植物园,广东深圳518000 [2]深圳出入境检验检疫局,广东深圳518045 [3]广东生态工程职业学院,广东广州510520

出　　处：《广东农业科学》2018年第7期100-104,173,共6页Guangdong Agricultural Sciences

基　　金：国家质检总局科技计划项目(2015IK267;2014IK008)

摘　　要：基于基质辅助激光解吸/电离飞行时间质谱(MALDI-TOFMS),对样品处理过程中的各条件进行优化,建立了基质辅助激光解吸/电离飞行时间质谱快速鉴定炭疽菌的方法。炭疽病菌菌丝采用70%甲酸:100%乙腈(1∶1)进行提取、50μL70%甲酸∶100%乙腈(1∶1)重悬后再超声提取,然后以HCCA为基质,乙腈∶三氟乙酸∶无菌去离子水(50%∶2.5%∶47.5%)为基质溶剂,运用改进的干滴法点样,进行MALDI-TOF-MS分析。应用此方法获得了咖啡浆果炭疽病菌特有的9个蛋白图谱峰,建立12种炭疽菌的蛋白指纹图谱库,从而建立MALDITOFMS分析炭疽菌的标准方法。In this study,we developed the method of identification Colletotrichum with Matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDl-TOFMS)and investigated its influencing factors.The mycelium was extracted by ultrasonic method in 50μL of 70%formic acid:100%acetonitrile(1:1).Then,used HCCA as matrix,acetonitrile/trifluoroacetic acid/sterile deionized water(Volume ratio was 50%:2.5%:47.5%)as matrix solution,and the improved dry drop method was used for MALDI-TOF-MS analysis.9 protein peaks of Colletotrichum kahawae were obtained by this method,and the protein fingerprint database of 12 species of Colletotrichum established,which could be used to distinguish different species.The method is simple,rapid,high throughput and high sensitivity,which can be used for rapid identification of Colletotrichum in the field of plant quarantine.

关 键 词：基质辅助激光解析/电离飞行时间质谱 炭疽菌 鉴定 蛋白指纹图谱 

分 类 号：S41-30[农业科学—植物保护]