四川农业大学学报 ›› 2006, Vol. 24 ›› Issue (04): 455-458.doi: 10.16036/j.issn.1000-2650.2006.04.020

• 研究简报 • 上一篇    下一篇

超声波处理对根癌农杆菌介导转化金钗石斛影响的初步研究

杨翠芹1, 秦耀国2, 刘伟3   

  1. 1. 四川农业大学 农学院, 四川 雅安 625014;
    2. 四川农业大学 林学园艺学院, 四川 雅安 625014;
    3. 华南农业大学 生命科学学院, 广东 广州 510642
  • 收稿日期:2006-04-13 出版日期:2006-12-31 发布日期:2017-03-06
  • 基金资助:
    广东省自然科学基金;广州市农业科技招标项目

Preliminary Study on Effects of Sonication on Agrobacterium-mediated Transformation of Dendrobium nobile Lindl

YANG Cui-qin1, QIN Yao-guo2, LIU Wei3   

  1. 1. College of Agriculture, Sichuan Agricultural University, Yaan 625014, Sichuan, China;
    2. College Forestry and Horticulture, Sichuan Agricultural University, Yaan 625014, Sichuan, China;
    3. College of Life Science, South China Agricultural University, Guangzhou 510642, Guangdong, China
  • Received:2006-04-13 Online:2006-12-31 Published:2017-03-06

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:25 KHz超声波40~80 W处理5~10 min或100 W处理5 min对PLBs死亡率无不良影响;1/2 MS液体培养基适宜作为超声溶液;80~100 W超声波处理5min的PLBs在转化后GUS染色每100 mg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。

关键词: 金钗石斛, 农杆菌, 转化, 超声波

Abstract: 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.

Key words: Dendrobium nobile Lindl, Agrobacterium, transformation, sonication

中图分类号: 

  • Q943.2