›› 2012, Vol. 20 ›› Issue (5): 921-926.DOI: 10.11733/j.issn.1007-0435.2012.05.020

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Regeneration Systems for in Vitro Culture of Filament and Genetic Variation Analyses of Yucca gloriosa L.

ZHAO Yong-qin1, ZHANG Na1, WU Xin-xin1, ZHENG He2, LI Hong-bo2, ZHAO Bing1, GUO Yang-dong1   

  1. 1. College of Agriculture and Biotechnology, China Agricultural University, Beijing 100193, China;
    2. Institute of Haidian Agricultural Science, Beijing 100080, China
  • Received:2012-03-15 Revised:2012-05-15 Online:2012-10-15 Published:2012-11-01

凤尾兰花丝诱导的组培再生体系的建立及遗传稳定性分析

赵永钦1, 张娜1, 吴新新1, 郑禾2, 李洪波2, 赵冰1, 郭仰东1   

  1. 1. 中国农业大学农学与生物技术学院, 北京 100193;
    2. 北京市海淀区农业科学研究所, 北京 100080
  • 通讯作者: 郭仰东,E-mail:yaguo@cau.edu.cn
  • 作者简介:赵永钦(1981-),男,河北沧州人,博士研究生,研究方向为植物生物技术,E-mail:zhaoyongqin2010cau@163.com
  • 基金资助:
    国家重大基础研究项目(2009CB119000); 中央高校基本科研业务费(2009-2-06)资助

Abstract: Using the filament as explants, the effects of different combinations of hormone on callus induction and differentiation, and the effects of activated carbon concentration on callus browning rate in tissue culture process of Yucca gloriosa L. were studied. Results showed that explanted filament could not only gain a higher propagation coefficient, but also reduce tissue damage from disinfectant contaminations. The best callus induction medium was MS+6-BA 5.0 mg稬-1+KT 1.0 mg稬-1+2,4-D 0.5 mg稬-1; and the best callus differentiation medium was MS+6-BA 6.0 mg稬-1+KT 0.5 mg稬-1 +NAA 0.5 mg稬-1; and the best in vitro shoot proliferation culture medium was MS+6-BA 3.0 mg稬-1 +KT 0.5 mg稬-1+NAA 0.05 mg稬-1+coconut milk 100 mL稬-1. Regenerated plantlets were tested by randomly amplified polymorphic DNA (RAPD) with no genetic variation detected. Regenerated plantlets can maintain genetic stability. This is the first report for in vitro culture and regeneration using filaments from Y. gloriosa L.

Key words: Yucca gloriosa L., Filament, Tissue culture, Genetic variation analysis, RAPD

摘要: 以凤尾兰(Yucca gloriosa L.)花丝为外植体,研究不同激素种类和配比对愈伤组织诱导和分化的影响,以及活性炭对外植体褐化率的影响,以期为进一步工厂化育苗提供技术支持。结果表明:以花丝为外植体不仅可以获得较高的繁殖系数,而且可以避免灭菌剂对外植体的伤害,从而有效降低组织培养的难度和污染率;凤尾兰花丝最佳愈伤诱导培养基为:MS+6-BA 5.0 mg·L-1 +KT 1.0 mg·L-1 +2,4-D 0.5 mg·L-1,最佳分化培养基为:MS+6-BA 6.0 mg·L-1 +KT 0.5 mg·L-1 +NAA 0.5 mg·L-1,最佳不定芽增殖培养基为:MS+6-BA 3.0 mg·L-1+KT 0.5 mg·L-1 +NAA 0.05 mg·L-1 +椰乳100 mL·L-1,增殖系数达到3.5左右;利用RAPD分子标记技术,在DNA水平上对再生株进行遗传变异鉴定,未检测出遗传变异,表明再生植株能够保持遗传上的稳定性。本研究首次建立了凤尾兰花丝高效稳定的组培再生体系。

关键词: 凤尾兰, 花丝, 组织培养, 遗传变异分析, RAPD

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