【摘要】 小麦(Triticum aestvum)纹枯病作为一种常见的土传真菌病害，已成为制约小麦产量的重要因素。本研究以禾谷丝核菌(Rhizoctonia cerealis)为研究对象，以聚乙二醇(polyethylene glycol, PEG)介导的遗传转化体系，获得带有GFP标记的能够稳定遗传的转化子；将该转化子侵染小麦，通过徒手切片观察，明确了禾谷丝核菌在小麦中的定殖规律。研究发现，通过PEG介导将携带GFP的质粒转入原生质体，并在含有100 mg/mL潮霉素的PDA培养基中进行筛选，获得了阳性转化子。经PCR检测及荧光观察，表明携带GFP的质粒已成功转入禾谷丝核菌中。经过连续传代培养，阳性转化子能够稳定表达，且其致病性与野生型一致。在小麦接种转化子后第4天，可在小麦根部木质部观察到明显荧光，第7天在小麦胚芽鞘韧皮部也可观察到明显荧光。本研究成功获得了带有GFP标记且具有良好遗传稳定性的禾谷丝核菌，并初步阐明了禾谷丝核菌在小麦中的定殖规律，为后续深入研究禾谷丝核菌的致病机制及利用微生物菌剂防治纹枯病提供了理论依据。更多还原

【Abstract】 As a common soil-borne fungal disease, wheat sharp eyespot has emerged as a crucial factor constraining wheat(Triticum aestvum) yield. In this study, a polyethylene glycol(PEG)-mediated genetic transformation system was employed to transform green fluorescent protein(GFP) into Rhizoctonia cerealis and obtained a transformant that was genetically stable; The GFP-labeled R. cerealis was inoculated to wheat seedlings, and the colonization pattern of R. cerealis in wheat was investigated by section observation. It was found that positive transformants were attained by PEG-mediated transformation onto protoplasts and screened in PDA medium containing 100 mg/mL hygromycin. PCR detection and fluorescence observation indicated that the plasmid carrying GFP had been successfully transfected into the R. cerealis. After successive passaging culture, the positive transformants were able to express stably and their pathogenicity was consistent with that of the wild type. On the fourth day after inoculating the transformants to wheat, distinct fluorescence could be observed in the xylem of the wheat root, and on the seventh day, a fluorescence phenomenon emerged in the phloem of the wheat coleoptile. This experiment successfully obtained R. cerealis with GFP labeling which could be stably inherited and preliminarily elucidated the colonization pattern of R. cerealis in wheat, it provide theoretical basis for further in-depth research on the pathogenic mechanism of R. cerealis and the use of microbial fertilizers to control wheat sharp eyespot.更多还原