Mandelc Stanislav, Javornik Branka
Biotechnical Faculty, University of Ljubljana, Ljubljana, Slovenia.
Proteomics. 2015 Feb;15(4):787-97. doi: 10.1002/pmic.201400181. Epub 2015 Jan 7.
Verticillium albo-atrum is a vascular wilt pathogen capable of infecting many important dicotyledonous plant species. Fungal isolates from hop differ in aggressiveness, causing either mild or lethal symptoms in infected plants. As in other plant pathogenic fungi, extracellular proteins, such as cell wall-degrading enzymes and effectors, are thought to be crucial in the pathogenesis process. In this study, mild and lethal isolates from three countries were grown in simulated xylem medium and secretome analysis by 2D-DIGE showed low qualitative and high quantitative variability among the isolates. Functional classification of 194 identified proteins representing 100 unique protein accessions revealed an arsenal of cell wall-degrading enzymes and potential effectors. The set of proteins that were more abundant in at least two lethal isolates included enzymes acetylcholinesterases, lipases, polygalacturonases, pectate lyase, rhamnogalacturonan acetylesterases, acetylxylan esterase, endoglucanase, xylanases, mannosidases, and a protein similar to alginate lyase and also potential effectors necrosis- and ethylene-inducing protein, small basic 14 kDa hypothetical protein and 79 kDa hypothetical proteins. Other proteins associated with virulence showed different expression profiles between mild and lethal isolates. The results suggest that the increased virulence of lethal isolates has little background shared by all three lethal isolates and that upregulation of isolate specific sets of proteins may be most important.
黑白轮枝菌是一种维管束萎蔫病原体,能够感染许多重要的双子叶植物物种。来自啤酒花的真菌分离株在侵袭性方面存在差异,在受感染的植物中会引起轻微或致命的症状。与其他植物病原真菌一样,细胞外蛋白,如细胞壁降解酶和效应子,被认为在致病过程中至关重要。在本研究中,从三个国家分离得到的温和型和致死型菌株在模拟木质部培养基中培养,通过二维差异凝胶电泳进行的分泌蛋白质组分析表明,这些分离株之间在定性方面差异较小,但在定量方面差异较大。对代表100个独特蛋白质登录号的194种已鉴定蛋白质进行功能分类,发现了一系列细胞壁降解酶和潜在的效应子。在至少两种致死型分离株中含量更高的蛋白质包括乙酰胆碱酯酶、脂肪酶、多聚半乳糖醛酸酶、果胶酸裂解酶、鼠李半乳糖醛酸乙酰酯酶、乙酰木聚糖酯酶、内切葡聚糖酶、木聚糖酶、甘露糖苷酶,以及一种类似于藻酸盐裂解酶的蛋白质,还有潜在的效应子坏死和乙烯诱导蛋白、小的碱性14 kDa假设蛋白和79 kDa假设蛋白。其他与毒力相关的蛋白质在温和型和致死型分离株之间表现出不同的表达谱。结果表明,致死型分离株毒力的增强在所有三种致死型分离株中几乎没有共同的背景,并且分离株特异性蛋白质组上调可能最为重要。