Establishment of an Agrobacterium ‐Mediated Genetic Transformation System for Prunus sibirica and Its Preliminary Application in

ABSTRACT

Prunus sibirica possesses substantial ecological and economic value and is recognized as an emerging woody energy crop; however, its regeneration and genetic transformation remain challenging. The present study established an efficient regeneration and Agrobacterium tumefaciens ‐mediated transformation system for P. sibirica . Stem segments served as explants, and the optimal proliferation medium was Murashige and Skoog medium supplemented with 0.8 mg L ⁻¹ 6‐benzylaminopurine (6‐BA), 0.2 mg L ⁻¹ thidiazuron (TDZ), and 0.1 mg L ⁻¹ 1‐naphthaleneacetic acid (NAA), resulting in a proliferation coefficient of 6.53. The genetic transformation system utilized A. tumefaciens strain GV3101 carrying reporter vectors pRI101‐GFP or pCAMBIA1301‐GUS, with optimal parameters including a 3‐day preculture, infection at OD ₆₀₀  = 0.6 for 20 min, and a 2‐day coculture. Transformants were selected in a medium containing 20 mg L ⁻¹ kanamycin and 300 mg L ⁻¹ cefotaxime, achieving an average transformation efficiency of 11.27%. As an initial application, PsWRKY17 ‐overexpressing lines were generated, exhibiting transcript levels 7–18 times higher than those in the empty‐vector control. Following low‐temperature treatment, PsWRKY17 ‐OE lines demonstrated increased SOD and POD activities and reduced relative electrical conductivity and MDA content compared to the control ( p  < 0.01). These results provide preliminary physiological evidence that PsWRKY17 overexpression is associated with altered low‐temperature stress responses in P. sibirica . However, the regulatory mechanism of PsWRKY17 requires further investigation. In summary, an effective regeneration and genetic transformation system for P. sibirica was developed, providing a foundation for future research and biotechnological applications.