Fernández-Acero Francisco Javier, Jorge Inmaculada, Calvo Enrique, Vallejo Inmaculada, Carbú María, Camafeita Emilio, López Juan Antonio, Cantoral Jesús Manuel, Jorrín Jesús
Laboratory of Microbiology, Marine and Environmental Sciences Faculty, University of Cádiz, Cádiz, Spain.
Proteomics. 2006 Apr;6 Suppl 1:S88-96. doi: 10.1002/pmic.200500436.
Botrytis cinerea is a phytopathogenic fungi causing disease in a number of important crops. It is considered a very complex species in which different populations seem to be adapted to different hosts. In order to characterize fungal virulence factors, a proteomic research was started. A protocol for protein extraction from mycelium tissue, with protein separation by 2-DE and MS analysis, was optimised as a first approach to defining the B. cinerea proteome. Around 400 spots were detected in 2-DE CBB-stained gels, covering the 5.4-7.7 pH and 14-85 kDa ranges. The averages of analytical and biological coefficients of variance for 64 independent spots were 16.1% and 37.5%, respectively. Twenty-two protein spots were identified by MALDI-TOF or ESI IT MS/MS, with some of them corresponding to forms of malate dehydrogenase and glyceraldehyde-3-phosphate dehydrogenase. Two more spots matched a cyclophilin and a protein with an unknown function.
灰葡萄孢是一种可在多种重要作物上引发病害的植物病原真菌。它被认为是一个非常复杂的物种，其中不同的种群似乎适应不同的宿主。为了表征真菌的毒力因子，开展了一项蛋白质组学研究。作为定义灰葡萄孢蛋白质组的首要方法，优化了从菌丝体组织中提取蛋白质、通过二维电泳进行蛋白质分离以及质谱分析的方案。在二维电泳考马斯亮蓝染色凝胶中检测到约400个斑点，覆盖5.4 - 7.7的pH范围和14 - 85 kDa的分子量范围。64个独立斑点的分析变异系数和生物学变异系数的平均值分别为16.1%和37.5%。通过基质辅助激光解吸电离飞行时间质谱或电喷雾离子阱串联质谱鉴定出22个蛋白质斑点，其中一些对应于苹果酸脱氢酶和甘油醛-3-磷酸脱氢酶的不同形式。另外两个斑点与一种亲环蛋白和一种功能未知的蛋白质相匹配。
