地黄高效叶片离体再生系统的建立  

Establishment of in vitro High Frequency Regeneration System of Rehmannia glutinosa Leaves

作　　者：王丰青[1,2] 田云鹤[1] 周勋[1] 李振方[2] 杜家方[1] 郑红艳[2] 张重义[1,2] 

机构地区：[1]河南农业大学中药材研究所,河南郑州450002 [2]福建农林大学中药材GAP研究所,福建福州350002

出　　处：《中国现代中药》2013年第2期122-126,共5页Modern Chinese Medicine

基　　金：国家自然科学基金(81274022;30973875);高等学校博士学科点专项科研基金资助项目(20123515110005);河南农业大学博士科研启动基金(30500509)

摘　　要：目的:通过分析不同浓度激素对地黄品种金九的愈伤诱导、再生芽分化等方面的影响,以期建立高效的地黄叶片再生系统,为地黄的快速繁殖和遗传转化研究奠定基础。方法:分析在添加不同浓度NAA、6-BA情况下地黄叶片的愈伤诱导率、再生芽分化率、再生芽生根率和增殖系数。结果:在多个激素浓度组合下均具有较高的叶片愈伤诱导率。最适于金九的叶片再生芽分化的激素浓度为NAA0.1mg·L-1、6-BA2.0mg·L-1。在适宜的激素浓度范围内,金九的再生芽分化率很高,最高可达96.67%。添加0.1mg·L-1的NAA有利于再生地黄植株壮苗和提高生根率。在含有NAA0.1mg·L-1、6-BA0.5mg·L-1的MS培养基上培养,则有利于再生芽的增殖。结论:NAA、6-BA的浓度对叶片再生芽分化、生根和增殖有显著影响。金九可作为地黄的分子生物学研究和遗传转化受体的良好材料。Objective：An appropriate conservation method for callus of Gentiana macrophylla Pall.slowly growing,and the relationship of physiological index and morphological index of the conserved callus were studied.Methods：To conserve callus,growth inhibitors and high permeability compounds were added to the medium,or different concentration of basic medium were used.After the callus was conserved for six month at room temperature,the morphological index was recorded and physiological index of the conserved callus of Gentiana macrophylla with different concentration of mannitol,CCC and sucrose were measured.Results：A large number of embryoid were generated in the surface of callus with the treatment of 1.0 mg/L ABA and 9% sucrose and grew well.The callus showed yellow-green and grew well under the treatment with 5% mannitol and PEG.Callus in PP333 grew commonly and callus in CCC was aging.Under the treatment with 9% sucrose,soluble sugar,starch and soluble protein content were significantly higher than treated with the other concentration of sucrose,mannitol or CCC.Conclusion：The best condition for callus conservation of Gentiana macrophylla was 1.0 mg/L ABA and 9% sucrose,followed by 5% mannitol and PEG.Soluble sugar,starch and soluble protein can partly reflect the survival rate of the callus conservation slowly growing.

关 键 词：地黄 叶片 再生 激素 

分 类 号：R282.71[医药卫生—中药学]