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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。
In order to establish Agrobacterium tumefaciens-mediated genetic transformation system of Festuca arundinacea Schreb., The embryogenic callus of L.grass-type fescue Effects of Various Factors on Agrobacterium - mediated Transformation. The results showed that Agrobacterium tumefaciens mediated embryogenic callus transformation of Forsythia suspensa: The callus was pre-cultured for 3 days on medium containing 0.5 mg / L BAP and NAA, respectively. The OD 600 of the solution was 0.7, Time 15min; Agrobacterium pre-culture medium and suspension medium and co-culture medium added 100μmol / L acetosyringone; co-culture temperature 23 ℃, co-culture time 3d, co-culture medium pH 5.2. The results of different concentrations of hygromycin screening experiments showed that using low concentration (50mg / L) of hygromycin continuous screening, regeneration obtained the most number of transgenic plants, it is the most desirable screening method.