Technical Centre of APIQ, Shenzhen Entry-Exit Inspection & Quarantine Bureau, Shenzhen 518010, P.R. China.
Rapid Commun Mass Spectrom. 2009 Dec;23(23):3647-54. doi: 10.1002/rcm.4296.
Verticillium spp. have been listed by the European and Mediterranean Plant Protection Organization (EPPO) and China as plant quarantine pests. Although attempts have been made to develop a simple routine laboratory assay to detect these organisms, none are routinely used. We describe for the first time a robust assay for reliable identification of Verticillium spp. using protein fingerprinting data obtained by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS). Several sample preparation methods and matrices were investigated to improve mass spectra for the routine identification of six species of Verticillium spp.(Verticillium dahiliae, V. alboatrum, V. fungicola, V. nigrescens, and V. lecanii) by MALDI-TOF-MS. Using the optimized experimental method, we constructed a protein fingerprint database for six species of Verticillium and established a analysis criteria of log(Score). This MALDI-TOF-MS protocol should prove useful as a rapid and reliable assay for distinguishing different Verticillium spp.
青枯菌属已被欧洲和地中海植物保护组织（EPPO）和中国列为植物检疫性有害生物。尽管已经尝试开发一种简单的常规实验室检测方法来检测这些生物，但没有一种方法被常规使用。我们首次描述了一种使用基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF-MS）获得的蛋白质指纹图谱数据来可靠鉴定青枯菌属的稳健检测方法。研究了几种样品制备方法和基质，以改善质谱图，以便通过 MALDI-TOF-MS 对 6 种青枯菌属（大丽轮枝菌、V. alboatrum、V. fungicola、V. nigrescens 和 V. lecanii）进行常规鉴定。使用优化的实验方法，我们为 6 种青枯菌属构建了蛋白质指纹数据库，并建立了 log（Score）的分析标准。该 MALDI-TOF-MS 方案有望成为一种快速可靠的鉴定不同青枯菌属的方法。
