<p><i>Allium senescens</i> L. (<i>A. senescens</i>) is extensively cultivated in North Asia and Europe, particularly in northern China, for its medicinal, ornamental, and ecological restoration properties. However, the lack of a stable genetic transformation system for <i>A. senescens</i> impedes the production of transgenic plants and significantly restricts research progress in this area. This study employed orthogonal experimental methods to investigate multiple factors related to disinfection techniques, tissue culture, and genetic transformation. This study is the first to report the regeneration system for <i>A. senescens</i>, with the following key findings: (1) The upper half of the stem node from sterile leek seedlings is the optimal explant. Adventitious shoot induction was achieved at a 96.30% rate on MS medium with 2&#xa0;mg/L 6-BA and 0.15&#xa0;mg/L NAA at pH 5.8. (2) Rooting of adventitious shoots was achieved at a 93.00% rate on 1/2 MS medium with 0.5&#xa0;mg/L NAA. (3) For the adventitious bud culture medium, the optimal concentrations of hygromycin (Hyg) and cephalosporin (Cef) are 50&#xa0;mg/L and 400&#xa0;mg/L, respectively. For the adventitious root culture medium, they are 30&#xa0;mg/L and 300&#xa0;mg/L. (4) Adding 200 µmol/L acetosyringone (AS), with a 3-day pre-culture, an OD₆₀₀ of 0.8 for the bacterial solution, and a 15-minute infection time, significantly enhances genetic transformation efficiency. This study not only establishes a solid foundation for genetic transformation, enabling functional gene research and in-depth exploration of various biological issues related to <i>A. senescens</i>, but also promotes precise molecular breeding of <i>A. senescens</i>.</p>