Rubisco基因遗传转化铁皮石斛的条件筛选与优化

Screening and Optimization of Rubisco Genetic Transformation of Dendrobium officinale Kimura et Migo

为筛选与优化农杆菌介导的铁皮石斛遗传转化的条件,本研究以无菌的铁皮石斛原球茎作为转化的受体材料,通过农杆菌介导法导入Rubisco基因,研究了农杆菌菌液浓度、侵染时间、潮霉素浓度、特美汀浓度对遗传转化效率的影响,建立遗传转化体系。结果表明,菌液浓度OD_(600)值为0.9、侵染时间30 min、TMT浓度为50 mg/L、Hyg浓度为55 mg/L时,最终的遗传转化效率最高(12.5%),适合作为铁皮石斛原球茎的遗传转化条件,这为后续进一步的基因鉴定及功能研究提供科学数据。

In order to screen and optimize the conditions of Agrobacterium mediated genetic transformation of Dendrobium candidum.The sterile protocorm of Dendrobium officinale Kimura et Migo was used as the recipient material for transformation.We transformed the gene of Rubisco Agrobacterium-mediate transformation method,exploring the influence of agrobacterium concentration,infection time,the concentration of hygromycin,and temetine concentration on genetic transformation efficiency,and established an efficient genetic transformation system.The results showed that the OD_(600) value of bacterial solution concentration was 0.9 and the infection time was 30 minutes,the TMT concentration was 50 mg/L,and the Hyg concentration was 55 mg/L,the genetic transformation efficiency is the highest(12.5%),which is suitable for the genetic transformation of Dendrobium officinale,which lays a foundation for further gene identification and provide scientific data.