节点文献
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.
【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.
【Key words】 Plantago major L. ‘Giant Turkish.’; callus induction; plant regeneration; adventitious shoots;
- 【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