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大车前体外再生体系的建立和优化

Establishment and Optimization of in vitro Regeneration System for Plantago major L.

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【作者】 李平陈华李银心

【Author】 LI Ping~ 1,2, CHEN Hua~1 and LI Yin-Xin~ 11 Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of Botany, The Chinese Academy of Sciences, Beijing, 1000932 The Graduate School of the Chinese Academy of Sciences, Beijing, 100039

【机构】 中国科学院植物研究所光合作用与环境分子生理学重点实验室中国科学院植物研究所光合作用与环境分子生理学重点实验室 北京100093中国科学院研究生院北京100039北京100093北京100093

【摘要】 大车前不仅有很高的药用价值,在生态学研究方面也是重要模式植物。关于大车前的组培,目前报道很少。我们通过不定芽直接再生和愈伤组织诱导两种途径,建立了大车前(PlantagomajorL.‘GiantTurkish.’)的快速高效组培再生系统。完整的成熟种子培养在添加IAA和TDZ的MS培养基中,不经过愈伤的分化阶段,从子叶节的部位产生不定芽,直接不定芽的诱导频率达到100%。在0.2mgLIAA和1.0mgLTDZ作用下,培养4~5周后平均每个外植体产生再生芽的数目达到14.6个。对同一个外植体诱导得到的9株再生植株进行的RAPD检测表明,部分植株在DNA水平上发生了变异。以叶片作为外植体,在添加1.0mgLNAA的MS固体培养基中培养3周后,伤口处形成愈伤组织,产生愈伤的频率平均为98%。愈伤组织在添加4.0mgL6BA的MS固体培养基中分化得到再生芽,分化频率为25%,平均每块愈伤产生再生芽2.8个。两种途径得到的再生芽转到12MS培养基上均可生根、长成完整植株,小苗移栽到温室90%能够存活。

【Abstract】 Plantago major is not only used as medicinal herb but also an important model plant of ecology. Little work has been reported on the tissue culture of P. major. A reproducible system for direct shoot morphogenesis and callus induction of Plantago major L. ‘Giant Turkish.’ was described. Using seed as explants, the adventitious buds were obtained 4 to 5 weeks following incubation on MS medium supplemented with 0.2 mg/L IAA and 1.0 mg/L TDZ. The frequency of adventitious buds was as high as 100%. The average number of buds per explant was 14.6. Random amplified polymorphic DNA analysis on 9 regenerants indicated that somaclonal variation occurred at DNA level. Using leaves as explants, calli were easily induced on MS medium supplemented with 1.0 mg/L NAA 3 weeks following inoculation. The frequency of callus induction can be as high as 98%. On MS medium containing 4.0 mg/L 6-BA, 25% of calli differentiated and the mean number of buds per piece of callus was 2.8. The buds developed roots on 1/2 MS medium and formed plantlets, 90% of which survived when transplanted to greenhouse.

【基金】 国家高技术研究与发展项目资助(No.2003AA627010)。~~
  • 【文献出处】 生物工程学报 ,Chinese Journal of Biotechnology , 编辑部邮箱 ,2005年06期
  • 【分类号】S567.2;
  • 【被引频次】11
  • 【下载频次】183
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