【摘要】 以建立的苜蓿高频再生体系为基础,利用GUS染色组织分析法,研究了影响根癌农杆菌(AgrobacteriumtumefaciensConn)LBA4404菌株介导的苜蓿(MedicagosativaL.)遗传转化的若干因素,建立了苜蓿快速有效的遗传转化体系,获得了转BADH基因植株。苜蓿的不同基因型、附加乙酰丁香酮的浓度、预培养与否、菌液浓度、共培养时间和卡那霉素浓度等对转化频率都有一定影响。最高转化频率可达43.3%,抗性植株经PCR和PCR-Southernblot检测表明,目的基因已整合进苜蓿基因组中。更多还原

【Abstract】 Several factors affected genetic transformation of alfalfa (Medicago sativa L.) mediated by Agrobacterium tumefaciens were studied by GUS coloration analysis and a quick and effective protocol with optimized condition for genetic transformation of alfalfa was developed on the basis of high-frequency tissue culture regeneration system and the BADH transgenic plants were achieved. Different genetypes, the concentrations of acetosyringone, preculture or not, bacterial concentrations, the time of co-cultivation and the concentrations of kanamycin had some effect on transformation frequency. The highest transformation frequency was 43.3%. The PCR and PCR-Southern blotting analysis showed that the BADH gene was integrated into genome of the plants.更多还原