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| Study on the Colonization of Rhizoctonia cerealis in Wheat (Triticum aestvum) Using GFP Labeled Strains |
| ZHANG Jing-Jing, FU Jia-Jia, LIU Na, HAN Sheng-Fang, HOU Chun-Yan, WANG Dong-Mei* |
| State Key Laboratory of North China Crop Improvement and Regulation/Hebei Key Laboratory of Plant Physiology and Molecular Pathology/College of Life Sciences, Hebei Agricultural University, Baoding 071001, China |
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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.
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Received: 10 December 2024
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Corresponding Authors:
*dongmeiwang63@126.com
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