Departamento de Bioquímica y Biología Molecular, Universidad de La Laguna, La Laguna, Tenerife, Spain.
Proteomics. 2010 Aug;10(16):3020-34. doi: 10.1002/pmic.201000037.
The extracellular proteome, or secretome, of phytopathogenic fungi is presumed to be a key element of their infection strategy. Especially interesting constituents of this set are those proteins secreted at the beginning of the infection, during the germination of conidia on the plant surfaces or wounds, since they may play essential roles in the establishment of a successful infection. We have germinated Botrytis cinerea conidia in conditions that resemble the plant environment, a synthetic medium enriched with low molecular weight plant compounds, and we have collected the proteins secreted during the first 16 h by a double precipitation protocol. 2-D electrophoresis of the precipitated secretome showed a spot pattern similar for all conditions evaluated and for the control medium without plant extract. The proteins in 16 of these spots were identified by PMF and corresponded to 11 different polypeptides. Alternative determination of secretome composition by LC-MS/MS of tryptic fragments rendered a much larger number, 105 proteins, which included all previously identified by PMF. All proteins were functionally classified according to their putative function in the infection process. Key features of the early secretome include a large number of proteases, the abundance of proteins involved in the degradation of plant defensive barriers, and plenty of proteins with unknown function.
植物病原真菌的细胞外蛋白质组或分泌组被认为是其感染策略的关键要素。在这个集合中，特别有趣的成分是那些在感染开始时、在分生孢子在植物表面或伤口上萌发时分泌的蛋白质，因为它们可能在建立成功感染中发挥重要作用。我们已经在类似于植物环境的条件下萌发了 Botrytis cinerea 分生孢子，即在富含低分子量植物化合物的合成培养基中，并且我们已经通过双重沉淀方案收集了在前 16 小时分泌的蛋白质。沉淀的分泌组的 2-D 电泳显示出所有评估条件和没有植物提取物的对照培养基相似的斑点模式。这些斑点中的 16 个斑点中的蛋白质通过 PMF 进行了鉴定，对应于 11 个不同的多肽。通过胰蛋白酶片段的 LC-MS/MS 对分泌组组成的替代测定产生了更多数量的 105 种蛋白质，其中包括所有通过 PMF 鉴定的蛋白质。所有蛋白质根据其在感染过程中的潜在功能进行了功能分类。早期分泌组的主要特征包括大量蛋白酶、参与植物防御屏障降解的蛋白质的丰富度以及大量具有未知功能的蛋白质。
