Abstract Pinus koraiensis holds the designation of a second-class protected wild plant in China, valued for its significant economic and ecological importance. However, the lack of a well-established genetic transformation system has hindered progress in functional research and breeding applications for this species. This study aimed to establish a reliable and efficient genetic transformation system of Pinus koraiensis embryonic callus, building upon somatic embryogenesis technology. Utilizing Pinus koraiensis embryonic callus and GUS as the reporter gene, Agrobacterium-mediated transformation was employed to investigate key transformation factors, such as antibiotic type and concentration, Agrobacterium bacterial solution concentration, infiltration, and co-cultivation times. The results revealed that 10 mg·L-1 of Hygromycin (Hyg) significantly inhibited Pinus koraiensis embryonic callus proliferation, while an OD600 absorbance value of 0.6 during transformation led to a remarkable 93.42±2.13% efficiency. Optimal co-cultivation for two days resulted in a transformation rate of 82.61%, with a high GUS staining rate of 88.89% in the resistant embryonic callus. Following the optimized protocol, resistant somatic embryos were successfully obtained. This research contributes to the advancement of seed resource breeding and genetic enhancement for Pinus koraiensis, providing a solid foundation for investigating gene functions related to this species.
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Hou et al. (2024) studied this question.
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