黄衡宇,王美蓉.紫背金盘愈伤组织诱导与再生体系的建立[J].中草药,2014,45():
紫背金盘愈伤组织诱导与再生体系的建立
Callus induction and regeneration system establishment of Ajugae nipponensis
  
DOI:
中文关键词:  紫背金盘  花序  愈伤组织  诱导  再生
英文关键词:Ajugae nipponensis Makino  inflorescence  callus  induction  regeneration
基金项目:湖南省高校科技创新平台资助项目(11K053)
作者单位
黄衡宇,王美蓉 云南中医学院 中药材优良种苗繁育工程研究中心云南 昆明 650500 
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中文摘要:
      目的 建立紫背金盘Ajugae nipponensis愈伤组织诱导体系,筛选出具较高再生率的植株发生途径,旨在构建高效、稳定的再生体系。方法 MS为基本培养基,添加不同种类和浓度配比的植物生长调节剂,以茎尖、带芽茎段和花序为外植体进行实验。结果 花序是较适宜诱导愈伤组织的外植体,在MS+6-BA 0.1 mg/L+2, 4-D 1.5 mg/L中可在1周内诱导出愈伤组织,2周后即分化出绿色小芽丛;丛生芽增殖的适宜培养基为MS+6-BA 2.0 mg/L+NAA 0.5 mg/L,愈伤组织芽丛再生率高达100%,再生系数为4.10,4周后增殖倍数5.0以上;生根的适宜培养基为1/2 MS+NAA 1.0 mg/L,3周后即可获得再生植株,生根率100%;生根苗移栽至排水良好的沙土中,成活率100%。结论 紫背金盘不同外植体均可诱导出愈伤组织,其中花序愈伤发生率最好,是最适宜的外植体;本研究为保护紫背金盘野生资源和发展人工栽培奠定了良好基础,也为遗传转化研究提供重要的科学依据。
英文摘要:
      Objective An efficient and stable regeneration system of Ajugae nipponensis was developed by establishing the callus induction system and selecting the best protocol for higher plant regeneration. Methods Stem tip, stem with buds, and inflorescence were all used as explants maintained on Murashige and Skoog (MS) medium supplemented with different types and concentration of plant growth regulator combinations. Results Inflorescence was the optimal explant for callus induction and was cultured on MS medium containing 0.1 mg/L 6-BA and 1.5 mg/L 2, 4-D in one week; The callus divided into green multiple shoot two weeks later; The optimal medium for multiple shoot proliferation was MS + 2.0 mg/L 6-BA + 0.5 mg/L NAA, the regeneration rate of callus clustered buds was as high as 100%, the regeneration coefficient was 4.10, and the proliferation increased by more than five times after four weeks; The optimal medium for rooting is 1/2 MS + 1.0 mg/L NAA, the plant regeneration occurred three weeks later and the rooting rate up to 100%; 100% of plantlet survived after transplanting rooted plantlets into sand. Conclusion The callus can be induced by all kinds of explants and inflorescence is the best explant for higher callus induction rate; The current study laid a solid foundation for protecting the wild resources and advancing the cultivated of A. nipponensis, meanwhile providing scientific evidence for the research of genetic transformation.
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