{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T09:09:47Z","timestamp":1789031387654,"version":"build-2803163510"},"reference-count":27,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T00:00:00Z","timestamp":1788998400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Wheat Science and Technology Innovation Team","award":["2025KJCXTD001"],"award-info":[{"award-number":["2025KJCXTD001"]}]},{"name":"Key Discipline Construction Fund for Crop Science of Anhui Science and Technology University","award":["XK-XJGF001"],"award-info":[{"award-number":["XK-XJGF001"]}]},{"award":["XK-XJGF001"],"award-info":[{"award-number":["XK-XJGF001"]}],"id":[{"id":"https:\/\/ror.org\/01pn91c28","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Innovation Project of Bio-breeding Laboratory of Anhui Province","award":["2025SWYZ0310"],"award-info":[{"award-number":["2025SWYZ0310"]}]},{"name":"Innovation Project of Bio-breeding Laboratory of Anhui Province","award":["2025SWYZ0340"],"award-info":[{"award-number":["2025SWYZ0340"]}]},{"name":"Collaborative Innovation Programs of Anhui University","award":["GXXT-2023-103"],"award-info":[{"award-number":["GXXT-2023-103"]}]},{"award":["GXXT-2023-103"],"award-info":[{"award-number":["GXXT-2023-103"]}],"id":[{"id":"https:\/\/ror.org\/05th6yx34","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Science and Technology Plan Project","award":["2024NY-03"],"award-info":[{"award-number":["2024NY-03"]}]},{"name":"Science and Technology Plan Project","award":["2023tpt035"],"award-info":[{"award-number":["2023tpt035"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Plants"],"abstract":"<jats:p>Genetic transformation is a core technology for wheat molecular breeding. Transformation efficiency and genotype compatibility determine the pace of targeted genetic improvement. Mature embryos are readily accessible but are associated with low transformation efficiency and strong genotype dependence. Using mature wheat embryos as explants, we screened eight wheat varieties representing three ecological types to develop genotype-adapted tissue culture systems. Treatments A5 (4 mg\/L 2,4-D + 2 mg\/L Dicamba), A7 (4 mg\/L 2,4-D + 6 mg\/L Dicamba), and A8 (4 mg\/L 2,4-D + 8 mg\/L Dicamba) were optimal for callus induction in winter, semi-winter, and spring wheat, respectively. WF3 (a 1\/2 MS-based medium) was broadly applicable to callus differentiation, whereas IBA produced the highest rooting efficiency in regenerated plantlets. The optimal transformation conditions comprised ultrasound treatment (2.5 min) followed by vacuum infiltration (5 min), supplementation of the infection solution with PEG6000 (250 mg\/L) and Tween20 (0.01%), and an OD600 of 0.6\u20130.8. Transformation efficiencies using mature embryos of the semi-winter wheat varieties YX288 and ZM578 were 2.5% and 1.8%, respectively. Using this transformation platform, the dwarfing genes Rht12 in ZM578 and Rht8 in YX288 were targeted by CRISPR\/Cas9-mediated editing. Multiple single-base substitutions at the target sites were detected in ZM578 rht12 mutants, which exhibited significantly reduced height, increased tiller number, and a compact plant architecture. However, no successfully edited rht8 lines were obtained in YX288. These findings indicate that this study established a reliable mature-embryo transformation system for wheat and successfully validated gene editing in the semi-winter variety ZM578. The system provides a technical platform for the functional analysis of key genes and molecular breeding in bread wheat.<\/jats:p>","DOI":"10.3390\/plants15182766","type":"journal-article","created":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T08:07:41Z","timestamp":1789027661000},"page":"2766","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Establishment of Tissue Culture and Genetic Transformation Systems Using Mature Embryos of Bread Wheat"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-4110-9617","authenticated-orcid":false,"given":"Yiyang","family":"He","sequence":"first","affiliation":[{"name":"College of Agriculture, Anhui Science and Technology University\/Bio-Breeding Laboratory of Anhui Province, Chuzhou 233100, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"}]},{"given":"Yingrun","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Agriculture, Anhui Science and Technology University\/Bio-Breeding Laboratory of Anhui Province, Chuzhou 233100, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-6001-9052","authenticated-orcid":false,"given":"Jia","family":"Shi","sequence":"additional","affiliation":[{"name":"College of Agriculture, Anhui Science and Technology University\/Bio-Breeding Laboratory of Anhui Province, Chuzhou 233100, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"given":"Songyu","family":"Pang","sequence":"additional","affiliation":[{"name":"College of Agriculture, Anhui Science and Technology University\/Bio-Breeding Laboratory of Anhui Province, Chuzhou 233100, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"given":"Wenyang","family":"Li","sequence":"additional","affiliation":[{"name":"College of Agriculture, Anhui Science and Technology University\/Bio-Breeding Laboratory of Anhui Province, Chuzhou 233100, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"}]},{"given":"Chuanzhi","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Biology and Food Engineering, Suzhou University, Suzhou 234000, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Lantian","family":"Ren","sequence":"additional","affiliation":[{"name":"College of Agriculture, Anhui Science and Technology University\/Bio-Breeding Laboratory of Anhui Province, Chuzhou 233100, China"},{"name":"Anhui Engineering Research Center for Smart Crop Planting and Processing Technology, Chuzhou 233100, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8028-6413","authenticated-orcid":false,"given":"Zhaoshi","family":"Xu","sequence":"additional","affiliation":[{"name":"Institute of Crop Science, Chinese Academy of Agricultural Sciences (CAAS)\/National Key Facility for Crop GeneResources and Genetic Improvement, Key Laboratory of Biology and Genetic Improvement of Triticeae Crops, Ministry of Agriculture, Beijing 100081, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Dong","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Agriculture, Anhui Science and Technology University\/Bio-Breeding Laboratory of Anhui Province, Chuzhou 233100, China"},{"name":"College of Agriculture, Northwest A&F University\/National Key Laboratory of Crop Stress Resistance and Efficient Production, Xianyang 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9475-7877","authenticated-orcid":false,"given":"Jiacheng","family":"Zheng","sequence":"additional","affiliation":[{"name":"College of Agriculture, Anhui Science and Technology 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