农杆菌介导高羊茅遗传转化体系的建立

为建立农杆菌(Agrobacterium tumefaciens)介导高羊茅(Festuca arundinaceaSchreb.)遗传转化体系,以草坪型高羊茅品种凌志的胚性愈伤组织为受体材料,通过gus基因瞬时表达检测,研究了多种因素对农杆菌介导转化的影响。结果表明,农杆菌介导高羊茅胚性愈伤组织转化的适宜条件为:愈伤组织在含BAP和NAA各0.5mg/L的培养基上预培养3d;菌液浓度OD600值0.7,感染时间15min;农杆菌预培养基和悬浮培养基以及共培养基中添加100μmol/L的乙酰丁香酮;共培养温度23℃,共培养时间3d,共培养基pH值5.2。不同浓度潮霉素筛选效果试验表明,采用低浓度(50mg/L)潮霉素连续筛选,再生获得的转基因植株数最多,因此是最为可取的筛选方式。

ESTABLISHMENT OF Agrobacterium-MEDIATED TRANSFORMATION SYSTEM FOR TALL FESCUE

In order to establish Agrobacterium-mediated transformation system for tall fescue (Festuca arundinacea Schreb.), the effects of multiple factors on transient expression of gus gene were investigated by using embryogenic calli derived from mature seeds of turf-type cultivar `Barlexas' as recipient materials. The results showed that the appropriate conditions for the Agrobacterium-mediated transformation of the embryogenic calli were as follows: callus pre-culture for 3d on medium containing BAP and NAA 0.5mg/L; bacterial suspension OD_(600) 0.7, infection time 15min; addition of 100μmol/L acetosyringone to Agrobacterium pre-culture and suspension media and co-culture medium; co-culture temperature 23℃, co-culture time 3d, co-culture medium pH 5.2. It demonstrated that continual selection with 50mg/L hygromycin produced the most transgenic plants.