生物技术通报 ›› 2013, Vol. 0 ›› Issue (4): 63-68.

• 研究报告 • 上一篇    下一篇

紫花苜蓿组织培养体系的建立及其遗传转化

周晓馥 吕杰 苗璐 高峰 徐洪伟   

  1. 吉林师范大学植物资源科学与绿色生产吉林省重点实验室,四平 136000
  • 收稿日期:2012-11-08 修回日期:2013-04-22 出版日期:2013-04-22 发布日期:2013-04-22
  • 作者简介:周晓馥,女,博士,教授,硕士生导师,研究方向:植物基因工程;E-mail: xiaofuzhou@sohu.com
  • 基金资助:
    国家科技部农业科技成果转化资金项目(2010GB2B100118 ),吉林省科技发展计划项目(20100803),吉林省高技术产业发展项目[吉发改高技(2009) 633 号],吉林师范大学研究生创新科研计划资助项目[研创新(学)(201116 号)]

Establishment of Alfalfa Tissue Culture System and Research on Agrobacterium-mediated Transformation System in Alfalfa

Zhou Xiaofu, Lü Jie, Miao Lu, Gao Feng, Xu Hongwei   

  1. Key Laboratory of Jilin Province for Plant Resources Science and Green Production,Siping 136000
  • Received:2012-11-08 Revised:2013-04-22 Published:2013-04-22 Online:2013-04-22

摘要: 为建立高效的农杆菌介导的紫花苜蓿的遗传转化体系,以紫花苜蓿无菌苗叶片作为外植体,用8 种含有不同激素 浓度梯度的培养基对其进行愈伤组织及不定芽的诱导,结果表明,M7 培养基(MS+1 mg/L 2,4-D + 2 mg/L CH + 0.25 mg/L KT + 2 mg/L 琼脂)的愈伤组织诱导率最高,可达100% ;继代表现最好,为胚性愈伤组织;同时,继续培养后可直接分化出不定芽,分 化率达100%。结果表明,最适宜条件分别为卡那霉素(Kan) 筛选浓度为75 mg/L ;农杆菌菌液OD600 值为0.45-0.6 之间。

关键词: 紫花苜蓿, 组织培养, 高频再生, 遗传转化, 农杆菌, 卡那霉素, OD600

Abstract: In order to establish high-efficiency alfalfa genetic transformation system,the influence factors of Agrobacterium-mediated transformation system were studied. The callus and adventitious bud of alfalfa were induced by culturing leaves as explants in 8 different kind of medium,named M1-M8, respectively. The results showed that M7 medium(MS+1 mg/L 2,4-D + 2 mg/L CH + 0.25 mg/L KT + 2 mg/L agar) was the best one which can induced the much more ammount of callus and adventitious bud of alfalfa than others. The results showed that the most suitable conditions for kanamycin was 75 mg/L; OD600 of agrobactium concerntration value for 0.45-0.6.

Key words: Alfalfa, Tissue culture, High frequency regeneration, Genetic transformation, Agrobacterium, Kanamycin OD600