杂交枫香胚性愈伤组织遗传转化体系研究

目的杂交枫香是我国重要的用材和观赏树种资源，但其遗传转化体系尚未建立。建立杂交枫香遗传转化体系为杂交枫香性状改良和基因功能研究提供了方法。方法本研究基于杂交枫香高效的体细胞胚胎发生技术，用根癌农杆菌介导的遗传转化法对其胚性愈伤组织进行遗传转化，对潮霉素选择压、菌液浓度、侵染时间、共培养时间、以及共培养温度等影响因素采用...

Agrobacterium tumefaciens-mediated transformation of hybrid sweetgum embryogenic callus

ObjectiveHybrid sweetgum is an important timber and ornamental tree resources in China, but its genetic transformation system has not been established yet. Establishing genetic transformation system of hybrid sweetgum provides a useful approach for trait improvement and allows us to conduct a functional identification of gene in hybrid sweetgum.MethodBased on the efficient somatic embryogenesis of hybrid sweetgum, the embryogenic calluses were transformed by Agrobacterium tumefaciens-mediated genetic transformation. Factors influencing transformation were studied by orthogonal experiment, including hygromycin selective pressure, concentration of Agrobacterium, infection time, co-culture time and co-culture temperature.ResultResults showed that the minimum lethal concentration of hygromycin to embryogenic callus was 10 mg/L. The number of Gus positive spots was highest when bacterium solution (OD600) was 0.8, co-culture time was 3 d, infection time was 10 min, co-culture temperature was 25 ℃. The most transgenic positive resistant calluses were obtained when bacterium solution (OD600) was 0.2, co-culture time was 2 d, infection time was 10 min, co-culture temperature was 23 ℃. And the optimal treatment combination was 0.2 bacterium solution (OD600), 2 d co-culture time, 20 min infection time, 23 ℃ co-culture temperature.ConclusionA total of 210 positive transgenic callus were obtained by molecular identification. The genetic transformation system was established, which provides more feasible foundation for the transformation of broadleaf tree callus.