摘要
以金钗石斛的类原球茎体(PLBs)为受体材料,研究了侵染前材料不同创伤处理对农杆菌介导转化频率的影响。结果表明:25KHz超声波40～80W处理5～10min或100W处理5min对PLBs死亡率无不良影响;1/2MS液体培养基适宜作为超声溶液;80～100W超声波处理5min的PLBs在转化后GUS染色每100mg鲜重材料上的蓝色斑点数由常规方法处理的0～0.54提高到6.05～7.90,GUS基因瞬时表达频率得到提高;筛选两个月后多数PLBs的GUS基因仍能稳定表达。说明超声波处理能提高农杆菌介导转化金钗石斛PLBs的频率。
Effects of different treatment of protocorm-like bodies （PLBs） of Dendrobium nobile Lindl before infection on Agrobacterium-mediated transformation frequency were studied. It is showed that there is no bad effect on death rate of PLBs as treated with sonication of 25 KHz, 40-80 W and 5 - 10 minutes or 100 W and 5 minutes; 1/2 MS liquid culture medium is suited to be sonic solution; Numbers of blue spots per 100 mg flesh weight of PLBs is enhanced from 0- 0.54 of treatment with routine methods to 6.05-7.90 of treatment with sonication of 25 KHz, 80-100 W and 5 minutes, so GUS transient expression frequency Of PLBs is enhanced. GUS can express stably after selection of two months. These results indicate sonication can enhance Agrobacterium-mediated transformation frequency of PLBs of Dendrobium nobile Lindl.
出处
《四川农业大学学报》
CSCD
2006年第4期455-458,共4页
Journal of Sichuan Agricultural University
基金
广东省自然科学基金
广州市农业科技招标项目
关键词
金钗石斛
农杆菌
转化
超声波
Denclrobium nobile Lindl
Agrobacterium
transformation
sonication
