biostudies-literatureWang MNational Natural Science Foundation of ChinaE1253https://www.ebi.ac.uk/biostudies/studies/S-EPMC6100196biostudies-literatureUnknown23(6)Take-all, caused by <i>Gaeumannomyces tritici</i>, is one of the most important wheat root diseases worldwide, as it results in serious yield losses. In this study, <i>G. tritici</i> was transformed to express the hygromycin B phosphotransferase using a combined protoplast and polyethylene glycol (PEG)-mediated transformation technique. Based on a series of single-factor experimental results, three major factors-temperature, enzyme lysis time, and concentration of the lysing enzyme-were selected as the independent variables, which were optimized using the response surface methodology. A higher protoplast yield of 9.83 × 10⁷ protoplasts/mL was observed, and the protoplast vitality was also high, reaching 96.27% after optimization. Protoplasts were isolated under the optimal conditions, withMolecules (Basel, Switzerland)Optimization of Production Conditions for Protoplasts and Polyethylene Glycol-Mediated Transformation of <i>Gaeumannomyces tritici</i>.PMC610019631272074Wang MFeng JZhang JHan LZhang XWang LfalseOptimization of Production Conditions for Protoplasts and Polyethylene Glycol-Mediated Transformation of <i>Gaeumannomyces tritici</i>.Take-all, caused by <i>Gaeumannomyces tritici</i>, is one of the most important wheat root diseases worldwide, as it results in serious yield losses. In this study, <i>G. tritici</i> was transformed to express the hygromycin B phosphotransferase using a combined protoplast and polyethylene glycol (PEG)-mediated transformation technique. Based on a series of single-factor experimental results, three major factors-temperature, enzyme lysis time, and concentration of the lysing enzyme-were selected as the independent variables, which were optimized using the response surface methodology. A higher protoplast yield of 9.83 × 10⁷ protoplasts/mL was observed, and the protoplast vitality was also high, reaching 96.27% after optimization. Protoplasts were isolated under the optimal conditions, with2018-01-01T00:00:00Z2018 May2025-04-05T14:47:32.172Z2019-03-27T00:06:40ZS-EPMC61001962979497510.3390/molecules23061253