| 程颖慧1，王 颖2，杨晓朱3，章桂明2，冯建军2，汪 莹2.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蛆菌[J].广东农业科学,2018,45(7):100-104 |
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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蛆菌 |
| Identification of quarantine colletotrichum isolatesby matrix-assisted laser desorption /ionization time-of-flight mass spectrometry |
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| DOI：10.16768/j.issn.1004-874X.2018.07.016 |
| 中文关键词: 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 |
| 英文关键词: MALDl-TOFMS Colletotrichum identification protein fingerprint |
| 基金项目:国家质检总局科技计划项目（2015IK267，2014IK008） |
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| 中文摘要: |
| 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、50 μL 70%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MALDI TOFMS分析炭疽菌的标准方法。 |
| 英文摘要: |
| 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 50 μ μL 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. |
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