农杆菌介导东方百合“西伯利亚”遗传转化体系建立

以东方百合"西伯利亚"(Lilium seberia)为材料,建立了百合再生和农杆菌介导的遗传转化体系.采用LBA4404和GV3101菌株对鳞片来源的愈伤组织进行侵染,并以鳞片作为对比转化材料,两种菌株携带有相同的双元载体pCAMBIA1301.研究结果表明,农杆菌侵染愈伤组织外植体转化率较高;乙酰丁香酮(AS)的添加能大大提高根癌农杆菌转化百合的效率.另外,共培养时间、不同菌株均对转化效率有一定的影响.筛选后获得的抗性植株经PCR检测和GUS组织化学染色证实外源基因已经整合到基因组DNA中并获得表达,转化率为5%～8%.

Establishment of \%agrobacterium\%-mediated transformation system for Lilium \%seberia\%

A reproductive plant regeneration and an \%agrobacterium\%-mediated transformation system are developed for Lilium seberia . \%Agrobacterium\% strains LBA4404 and GV3101 are used, with each containing a binary vector pCAMBIA1301. Squama-derived calli are inoculated with strains, compared with the transformation material of squama. The results show that calli explants are more sensitive to \%agrobacterium\% infection in two different regeneration methods, induction of via gene with AS greatly improves the transformation effects. In addition, the time of co-cultivation, as well as different strains affects the transformation efficiency. Resistant plants can be obtained after transformtion and selective regeneration. GUS staining, PCR analysis confirm that transgene is integrated into the Lilium seberia\% genome with a transformation frequency of 5%~8%.