{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T06:05:03Z","timestamp":1790229903224,"version":"4.1.0"},"reference-count":66,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T00:00:00Z","timestamp":1790208000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:p>\n                    <jats:italic>Wilsonomyces carpophilus<\/jats:italic>\n                    is a fungal pathogen causing shot-hole disease in stone fruit trees, including wild apricot populations in the Tianshan wild fruit forests. The absence of an effective genetic transformation system has limited molecular studies of this pathogen. In this study, we established a PEG-mediated protoplast transformation system for\n                    <jats:italic>W. carpophilus<\/jats:italic>\n                    strain G052 5 m2. Key factors affecting protoplast preparation, including mycelial culture duration, enzyme combination, digestion time, and osmotic stabilizer, were evaluated. The optimal conditions consisted of 24-h-old mycelia, enzymatic digestion with 20 g\/L Driselase and 25 g\/L Snailase for 5 h, and 1.2 mol\/L NaCl as the osmotic stabilizer, yielding up to 4.13 \u00d7 10\n                    <jats:sup>6<\/jats:sup>\n                    protoplasts\/mL. The PEG-mediated transformation efficiency was calculated as 1.75 transformants per microgram of plasmid DNA under the optimized conditions. Both PCR detection and EGFP fluorescence observation successfully validated the exogenous gene expression in fungal transformants. No obvious differences in colony morphology, vegetative growth, and pathogenicity were observed between transformants and the wild-type strain. This study provides the first PEG-mediated protoplast transformation system for\n                    <jats:italic>W. carpophilus<\/jats:italic>\n                    and establishes a practical molecular tool for future studies on gene function, pathogenicity mechanisms, and host\u2013pathogen interactions in this important stone fruit pathogen.\n                  <\/jats:p>","DOI":"10.3389\/fmicb.2026.1917079","type":"journal-article","created":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T05:44:29Z","timestamp":1790228669000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["A PEG-mediated protoplast transformation system for EGFP labeling of the shot-hole pathogen Wilsonomyces carpophilus"],"prefix":"10.3389","volume":"17","author":[{"given":"Xinmei","family":"Zhou","sequence":"first","affiliation":[{"name":"College of Forestry and Landscape Architecture, Xinjiang Agricultural University","place":["Urumqi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chuli","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Forestry, Beijing Forestry University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiayuan","family":"Ouyang","sequence":"additional","affiliation":[{"name":"College of Forestry and Landscape Architecture, Xinjiang Agricultural University","place":["Urumqi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yiheng","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Forestry and Landscape Architecture, Xinjiang Agricultural University","place":["Urumqi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dianguang","family":"Xiong","sequence":"additional","affiliation":[{"name":"College of Forestry, Beijing Forestry University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chengming","family":"Tian","sequence":"additional","affiliation":[{"name":"College of Forestry, Beijing Forestry University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yusufjon","family":"Gafforov","sequence":"additional","affiliation":[{"name":"Department of Botany and Genetics, National University of Uzbekistan","place":["Tashkent, Uzbekistan"]},{"name":"Department of Medicine, Faculty of Medicine, Alfraganus University","place":["Tashkent, Uzbekistan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rong","family":"Ma","sequence":"additional","affiliation":[{"name":"College of Forestry and Landscape Architecture, Xinjiang Agricultural University","place":["Urumqi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,9,24]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"275","DOI":"10.5962\/p.416805","article-title":"Morphology and ontogeny of conidia in Wilsonomyces carpophilus, gen. nov. and comb. nov., causal pathogen of shot hole disease of Prunus species.","volume":"37","author":"Adaskaveg","year":"1990","journal-title":"Mycotaxon"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1099\/acmi.ac2019.po0012","article-title":"Studies on amylase from protoplast fusants of Aspergillus species using response surface methodology.","volume":"1","author":"Adeleye","year":"2019","journal-title":"Access. 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