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为优化金钗石斛的转化体系,以其类原球茎(PLBs)为材料,利用正交设计研究了影响农杆菌介导转化的因素.结果表明:PLBs预培养1 d、侵染的菌液OD600值1.2、乙酰丁香酮(AS)100μmol/L、共培养5 d时,每100 mg鲜物质量材料上的GUS染色蓝点数最多;单因子试验表明AS适宜的浓度为50μmol/L;PCR结果显示得到了转化呈阳性的PLBs;初步表明此体系可用于金钗石斛的遗传转化研究.
In order to optimize the transformation system of Dendrobium nobile, the factors affecting the Agrobacterium-mediated transformation were studied by orthogonal design using its protocorm-like bodies (PLBs) as experimental materials.The results showed that: PLBs were pre-cultured for 1 day, OD600 Value of 1.2, acetosyringone (AS) 100μmol / L, co-cultured for 5 d, the highest number of GUS staining blue spots per 100 mg fresh material quality materials; single factor test showed that AS suitable concentration of 50μmol / L; PCR results show The positive transformed PLBs were obtained. The results showed that this system could be used to study the genetic transformation of Dendrobium nobile.