乌饭树再生及遗传转化体系的建立

The Establishment of Regeneration and Genetic Transformation System for Vaccinium bracteatum Thunb

摘要: 本研究以乌饭树叶片为材料，筛选诱导叶片再生不定芽的植物生长调节剂种类和浓度，建立了乌饭树再生体系。此外，确定了用于筛选转基因材料的抗生素浓度、侵染乌饭树叶片的根癌农杆菌菌液浓度和侵染时间，建立了乌饭树遗传转化体系。结果表明：(1) 乌饭树叶片不定芽再生的最佳培养基配方为：WPM 0.5 mg/L TDZ 4 mg/L ZT；(2) 乌饭树叶片预培养5 d，筛选用抗生素潮霉素浓度为5 mg/L，农杆菌抑制剂头孢霉素浓度为350 mg/L，共培养6 d，农杆菌浓度为0.8，侵染时间为60 min时，遗传转化效率最高。上述结果为在乌饭树中进行基因功能验证和分子育种提供了理论基础和技术支撑。

Abstract: In this study, leaves of Vaccinium bracteatum Thunb were used as materials to screen the types and concentrations of plant growth regulators inducing adventitious bud regeneration, thus establishing the regeneration system of Vaccinium bracteatum Thunb. Additionally, optimal concentrations of antibiotics for screening transgenic materials, as well as concentrations and duration of Agrobacterium tumefaciens infection on Vaccinium bracteatum Thunb leaves, were determined to establish a genetic transformation system. The results revealed that: (1) the formula of adventitious bud regeneration medium of Vaccinium bracteatum Thunb leaves was 0.5 mg/L TDZ 4 mg/L ZT; (2) the pre-culture time of Vaccinium bracteatum Thunb leaves was 5 days, the optimal concentration of the antibiotic hygromycin for screening was 5 mg/L, the optimal concentration of the Agrobacterium inhibitor cephalosporin was 350 mg/L, the co-culture time was 6 days, the concentration of Agrobacterium was 0.8, and the infection time was 60 minutes, at which the infection efficiency was the highest. These findings provide a theoretical basis and technical support for gene function verification and molecular breeding in Vaccinium bracteatum Thunb.