Molecular and Cellular Biology, School of Science and Technology, University of New England, Armidale, NSW, Australia.
Proteomics. 2010 Apr;10(8):1573-91. doi: 10.1002/pmic.200900301.
Thielaviopsis basicola is a hemibiotroph fungus that causes black root rot disease in diverse plants with significant impact on cotton production in Australia. To elucidate how T. basicola growth and proteome are influenced by interactions with natural sources, this fungus was cultured in the presence of root extracts from non-host (wheat, hairy vetch) and susceptible host (cotton, lupin) plants. We found that T. basicola growth was significantly favored in the presence of host extracts, while hierarchical clustering analysis of 2-DE protein profiles of T. basicola showed plant species had a larger effect on the proteome than host/non-host status. Analysis by LC-MS/MS of unique and differentially expressed spots and identification using cross-species similarity searching and de novo sequencing allowed successful identification of 41 spots. These proteins were principally involved in primary metabolism with smaller numbers implicated in other diverse functions. Identification of several "morpho" proteins suggested morphological differences that were further microscopically investigated. Identification of several highly expressed spots suggested that vitamin B(6) is important in the T. basicola response to components present in hairy vetch extract, and finally, three spots, induced in the presence of lupin extract, may correspond to malic enzyme and be involved in lipid accumulation.
基腐隔孢腔菌是一种半活体真菌，可引起多种植物的黑根腐病，对澳大利亚的棉花生产有重大影响。为了阐明基腐隔孢腔菌的生长和蛋白质组如何受到与天然来源相互作用的影响，本研究在存在非宿主（小麦、野豌豆）和易感宿主（棉花、羽扇豆）植物根提取物的情况下培养了这种真菌。我们发现，基腐隔孢腔菌在存在宿主提取物的情况下生长明显受到促进，而基腐隔孢腔菌的 2-DE 蛋白质图谱的层次聚类分析表明，植物物种对蛋白质组的影响大于宿主/非宿主状态。通过 LC-MS/MS 对独特和差异表达斑点进行分析，并使用跨物种相似性搜索和从头测序进行鉴定，成功鉴定了 41 个斑点。这些蛋白质主要参与初级代谢，较少参与其他多种功能。鉴定出几种“形态”蛋白表明存在形态差异，进一步通过显微镜进行了研究。鉴定出几个高表达的斑点表明，维生素 B（6）在基腐隔孢腔菌对野豌豆提取物中存在成分的反应中很重要，最后，在羽扇豆提取物存在的情况下诱导了三个斑点，可能对应于苹果酸酶并参与脂质积累。
