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.

作　　者：李平[1] 陈华[1] 李银心[1] 

机构地区：[1]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

出　　处：《生物工程学报》2005年第6期916-922,共7页Chinese Journal of Biotechnology

基　　金：国家高技术研究与发展项目资助(No.2003AA627010)。~~

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。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 was2.8. The buds developed roots on 1/2 MS medium and formed plantlets, 90% of which survived when transplanted to greenhouse.

关 键 词：大车前(Plantago major L.'Giant Turkish.’) 愈伤诱导 植株再生 不定芽 

分 类 号：S567.2[农业科学—中草药栽培]