摘要
利用根癌农杆菌介导法将 GUS基因导入中国水仙 ,组织化学法检测转化后共培养 3d的水仙鳞茎盘组织块 ,GUS基因瞬时表达很好 ,90 %以上的材料呈现大面积的蓝色斑块 ,取筛选培养 2 0d的材料检测 GU S基因的稳定表达 ,约 1%的组织块有蓝色反应 .水仙预培养 10 d后转化 ,转化前在菌液中加入 60 μmol/ L AS活化 2~ 3h,转化率提高了两倍以上 .转化材料在 MS+ BA10 + NAA0 .2+ Cef30 0 + Hyg30 培养基上培养 2 0 d左右分化出小鳞芽 ,频率约为 60 % ,组织块平均出芽 5 .7个 ,最多可达 14个 ,小鳞芽 10 d后长成小鳞茎 .
The GUS gene was transformed into Narcissus tazetta mediated by Agrobacter ium tumefaciens . Bulbs of narcissus co cultivated 3 days after being transf ormed were detected by histochemical method, and GUS gene expressed perfectly. T here were large masses of blue patches on 90% of transformed materials. When sc reen cultured 20 days, about 1% tissues of the detected materials had long ter m expression of GUS gene. It indicated that GUS gene had been expressed steadil y. The frequency of transformation increased twice when narcissus pre cultured 10 days and agrobacterium was activated for 2~3 hours by 60 μmol/L AS before b eing transformed. Many bulbils outgrew from 60% of transformed materials which cultu red 20 days in MS+BA 10 +NAA 0.2 +Cef 300 +Hyg 30 medium, 5.7 bulbils on each tissue in average, and 14 at most.
出处
《厦门大学学报(自然科学版)》
CAS
CSCD
北大核心
2001年第5期1145-1150,共6页
Journal of Xiamen University:Natural Science
基金
福建省自然科学基金 (C970 0 1)
教育部访问学者基金资助项目