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[目的]优化蝴蝶兰和文心兰的遗传转化条件。[方法]以培养3周的蝴蝶兰原球茎和文心兰愈伤组织为试材,以大肠杆菌为生长培养基,研究3种农杆菌菌株的侵染力,以及菌液浓度、侵染时间、酚类诱导物(AS)和共培养时间对转化的影响。[结果]以蝴蝶兰原球茎为农杆菌介导的外植体进行遗传转化的优化条件为:预培养时间3d,菌液侵染浓度OD600=0.3,菌液侵染时间10min,pH值5.4,菌液侵染后,与农杆菌共培养5d并添加100μmol/LAS,此条件下的瞬间表达率最高。将蝴蝶兰遗传转化的条件运用于文心兰,得到的转化率较低。3种农杆菌菌株中,EHA105菌株侵染力最强,其次是AGL1,LBA4404最弱。[结论]该研究为进一步研究蝴蝶兰和文心兰的遗传转化体系提供理论依据。
[Objective] The research aimed to optimize genetic transformation conditions of Phalaenopsis and Oncidium. [Method] With 3-week-old Phalaenopsis protocormi callus and oncotic callus as tested material, E.coli was used as growth medium to study the infectivity of 3 kinds of Agrobacterium strains, as well as the bacterial liquid concentration, infection time, Effects of Phenolic Inducer (AS) and Coculture Time on Transformation. [Result] The optimal conditions for the transformation of Phalaenopsis by Agrobacterium tumefaciens were: pre-culture time 3d, OD600 = 0.3, bacterial infection time 10min, pH value 5.4, After inoculation, the cells were co-cultured with Agrobacterium for 5 days and added with 100μmol / LAS, the highest transient expression rate was obtained under these conditions. The application of the conditions for the genetic transformation of Phalaenopsis to Oncidium resulted in a lower conversion rate. Of the three Agrobacterium strains, EHA105 strain had the strongest infectivity, followed by AGL1, while LBA4404 was the weakest. [Conclusion] The study provided the theoretical basis for further study on the genetic transformation system of Phalaenopsis and Oncidium.