树状多节孢HQD_(33)遗传转化体系建立的影响因素研究

Factors influencing Agrobacterium tumefaciens-mediated transformation in Nodulisporium sylviforme

利用根癌农杆菌(Agrobacterium tum efaciens)介导的转化方法,以紫杉醇产生菌———树状多节孢HQD33的分生孢子为受体材料,分析影响转化效率的主要因素.获得高效的遗传转化体系,转化率最高可达1.3×10-4.PCR检测表明,T-DNA已整合进树状多节孢HQD33的基因组中,而且大多数都是以单拷贝的形式整合,转化子都能够稳定遗传.结果表明,根癌农杆菌菌株类型、菌株生长状态、分生孢子浓度、共培养时间以及乙酰丁香酮的诱导等因素对转化效率都具有重要影响.通过对以上因素的分析,为建立紫杉醇产生菌———树状多节孢HQD33稳定的遗传转化体系奠定了基础.

Agrobaderium tumefacens-mediated transformation system was used to study on factors influencing transformation in nodulisporium syluiforme HQD33.The results showed that the transformation efficiency was correlated with .4. tumefaciens strains, the growing condition of A. tumefaciens strains, concentration of conidia, cocultivation time and acetosyringone induction. Through setting up Agrobaderium tumefadens - mediated transformation system, the production of taxol will be increased greatly. These findings should facilitate future study of nodulisporium sylviforme HQD33.