蝴蝶兰·文心兰遗传转化体系的初步研究

Preliminary Study on the Genetic Transformation Systems of Phalaenopsis and Oncidium

[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 purpose was to optimize the genetic transformation condition of Phalaenopsis and Oncidium. [Method] With. Phalaenopsis protocorms and Oncidium calli cultured for 3 weeks as tested materials, Escherichia coli as growth medium, the infectivity of 3 agrobacterium strains, the effects of bacterium concn., infecting time, phenols inducer （AS） and co -culture time on the transformation were studied. [Result] With Phalaenopsis protocorms as explants for the agrobacterium-mediated transformation, the optimizing condition for genetic transformation was pre-culture time of 3 d, bacterium concn, of OD600=0.3, infecting time of 10 min and pH value of 5.4, the infected bacterium solution being co-cultured with agrobacteria for 5 d and added 100 μmol/L AS, under which the rate of instantaneous expression was highest. The transformation rate obtained by applying the genetic transformation condition of Phalaenopsis on Oncidium was lower. Among the 3 agrobacterium strains, the infectivity of strain EHA 105 was the strongest, that of AGL1 was secondary and that of LBA4404 was weakest. [Conclusion] The study provided theoretical foundation for further studying the genetic transformation systems of Phalaenopsis and Ortcidium.