Dai Tingting, Xu Yue, Yang Xiao, Jiao Binbin, Qiu Min, Xue Junxin, Arredondo Felipe, Tyler Brett M
Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, China.
Foreign Disease-Weed Science Research Unit, United States Department of Agriculture, Agricultural Research Service (ARS), Fort Detrick, MD, United States.
Front Microbiol. 2021 Jul 5;12:682754. doi: 10.3389/fmicb.2021.682754. eCollection 2021.
is a destructive pathogen causing root rot and dieback diseases on hundreds of economically and ecologically important plant species. Effective transformation systems enable modifications of candidate genes to understand the pathogenesis of . A previous study reported a polyethylene glycol and calcium dichloride (PEG/CaCl)-mediated protoplast transformation method of . However, the virulence of the transformants was compromised. In this study, we selected ATCC 15400 as a suitable wild-type isolate for PEG/CaCl transformation using the green fluorescent protein after screening 11 isolates. Three transformants, namely, PcGFP-1, PcGFP-3, and PcGFP-5, consistently displayed a green fluorescence in their hyphae, chlamydospores, and sporangia. The randomly selected transformant PcGFP-1 was as virulent as the wild-type isolate in causing hypocotyl lesions on lupines. Fluorescent hyphae and haustoria were observed intracellularly and intercellularly in lupine tissues inoculated with PcGFP-1 zoospores. The potential application of this improved transformation system for functional genomics studies of is discussed.
是一种具有破坏性的病原体，可导致数百种具有经济和生态重要性的植物物种发生根腐病和枝枯病。有效的转化系统能够对候选基因进行修饰，以了解其发病机制。先前的一项研究报道了一种聚乙二醇和二氯化钙（PEG/CaCl）介导的原生质体转化方法。然而，转化体的毒力有所受损。在本研究中，我们在筛选了11个分离株后，选择ATCC 15400作为适合使用绿色荧光蛋白进行PEG/CaCl转化的野生型分离株。三个转化体，即PcGFP-1、PcGFP-3和PcGFP-5，在其菌丝、厚垣孢子和孢子囊中始终显示出绿色荧光。随机选择的转化体PcGFP-1在引起羽扇豆下胚轴病变方面与野生型分离株的毒力相同。在用PcGFP-1游动孢子接种的羽扇豆组织中，在细胞内和细胞间观察到荧光菌丝和吸器。讨论了这种改进的转化系统在功能基因组学研究中的潜在应用。
