金钗石斛类原球茎转化子选择方式的比较

为确立农杆菌介导法转化金钗石斛类原球茎的适宜的选择体系,利用液体纸基、固体培养基与液体滤纸桥3种培养方式,以0,20,40,50,60,80 mg/L质量浓度的潮霉素(Hyg)梯度试验,发现不同培养方式的选择效果有所差异。当使用液体纸基培养在50 mg/L Hyg的选择压下,20 d后可使材料死亡;使用固体培养基时,60 mg/L Hyg的选择压下,1个月以上才见效;在40 mg/L的选择压下,液体滤纸桥选择需要26~29 d。固体培养基选择需要较高的Hyg质量浓度且选择周期长,液体纸基选择可缩短时间,但容易造成转化细胞的生长抑制甚至死亡,比较而言液体滤纸桥是较好的选择方式。

Comparison on Methods for Selection of theTransformed PLBs of D. nobile

To establish the appropriate system for selection of Agrobacterium-mediated transformation of protocorm-like bodies(PLBs)of Dendrobium nobile Lindl,materials were cultured in liquid paper medium,solid culture medium and liquid filter paper bridge,with Hyg at 7 different concentrations such as 0,20,40,50,60 and 80 mg/L.It was observed that different ways of selection had different effects.When using liquid paper medium selection with Hyg concentration of 50 mg/L,materials died in twenty days.It took more than one month to take effect with Hyg concentration of 60 mg/Lusing solid culture medium selection,and twenty-six to twenty-nine days were required on liquid filter paper bridge selection with 40 mg/L Hyg.Solid culture medium selection required a higher level of concentration and the selection cycle was long.Liquid paper mediumselection required a short time,but inhibited the growth of transformed cells or caused death.Liquid filter paper bridge was a good selection method.