Aberdeen Oomycete Laboratory, University of Aberdeen, Institute of Medical Sciences, Foresterhill, Aberdeen, UK.
Fungal Biol. 2012 Jan;116(1):24-41. doi: 10.1016/j.funbio.2011.09.004. Epub 2011 Sep 29.
The oomycete Pythium oligandrum is a mycoparasitic biocontrol agent that is able to antagonise several plant pathogens, and can promote plant growth. In order to test the potential usefulness of P. oligandrum as a biocontrol agent against late blight disease caused by the oomycete Phytophthora infestans, we investigated the interaction between P. oligandrum and Ph. infestans using the green fluorescent protein (GFP) as a reporter gene. A CaCl(2) and polyethylene-glycol-based DNA transformation protocol was developed for P. oligandrum and transformants constitutively expressing GFP were produced. Up to 56 % of P. oligandrum transformants showed both antibiotic resistance and fluorescence. Mycoparasitic interactions, including coiling of P. oligandrum hyphae around Ph. infestans hyphae, were observed with fluorescent microscopy. To gain further insights into the nature of P. oligandrum mycoparasitism, we sequenced 2376 clones from cDNA libraries of P. oligandrum mycelium grown in vitro, or on heat-killed Ph. infestans mycelium as the sole nutrient source. 1219 consensus sequences were obtained including transcripts encoding glucanases, proteases, protease inhibitors, putative effectors and elicitors, which may play a role in mycoparasitism. This represents the first published expressed sequence tag (EST) resource for P. oligandrum and provides a platform for further molecular studies and comparative analysis in the Pythiales.
卵菌纲的寡养单胞菌是一种真菌寄生性生物防治剂，能够拮抗几种植物病原体，并能促进植物生长。为了测试寡养单胞菌作为生物防治剂防治疫霉属卵菌引起的晚疫病的潜力，我们使用绿色荧光蛋白（GFP）作为报告基因研究了寡养单胞菌和疫霉属之间的相互作用。建立了一种基于氯化钙和聚乙二醇的 DNA 转化方案，用于转化寡养单胞菌，并产生了持续表达 GFP 的转化体。高达 56%的寡养单胞菌转化体表现出抗生素抗性和荧光性。通过荧光显微镜观察到寡养单胞菌菌丝缠绕在疫霉菌丝周围的真菌寄生相互作用。为了进一步了解寡养单胞菌真菌寄生的性质，我们从体外生长的寡养单胞菌菌丝或作为唯一营养源的热杀死的疫霉菌丝的 cDNA 文库中测序了 2376 个克隆。获得了 1219 个共识序列，包括编码葡聚糖酶、蛋白酶、蛋白酶抑制剂、假定效应子和激发子的转录本，它们可能在真菌寄生中发挥作用。这代表了寡养单胞菌的第一个发表的表达序列标签（EST）资源，为进一步的分子研究和在 Pythiales 中的比较分析提供了一个平台。
