农杆菌介导的百合ACO反义基因遗传转化的研究

Research on Genetic Transformation of ACO Antisense Gene into Lily Mediated by Agrobacterium Tumefaciens

以东方百合‘索邦’(Liliumoriental‘Sorbonne’)无菌苗鳞茎、鳞片叶及愈伤组织为外植体,通过EHA105和GV3101两种根癌农杆菌(Agrobacterium tumefaciens)菌株介导,将ACO反义基因导入东方百合"索邦"。结果表明,愈伤组织预培养7 d,侵染30 min,EHA105菌液浓度OD=0.8,共培养5 d,可获得7.14%的转化率。经GUS、PCR、PCR-Southern及基于梯状胶回收的PCR法检测证明ACO反义基因已转入转化植株基因组中。

In vitro bulbs, scale leaves and callus of Lilium oriental ‘Sorbonne＇ were used as the explants. ACO antisense gene had been transformed into Lilium oriental ‘Sorbonne＇ mediated by Agrobacterium turnefaciens EHA105 and GV3101. The results showed 7.14% of genetic transformation rate was obtained according to the procedures： the callus pre cultured 7 d, immersed into bacterial suspension （OD600=0.8） of EHA105 for 30 min, and then co-cultivated for 5 d. The ACO antisense gene was integrated into the genome of transgenic plants confirmed by PCR, GUS, PCR-Southern and ladderlike purification based PCR analysis.