Acta Agrestia Sinica ›› 2024, Vol. 32 ›› Issue (8): 2523-2530.DOI: 10.11733/j.issn.1007-0435.2024.08.019

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Establishing Tissue Culture and Regeneration System of Dracocephalum rupestre

YUE Kang-jie, KANG Hong-wei, LIU Hui-xin, WANG Jia-li, TIAN Xu-ping   

  1. College of Forestry, Shanxi Agricultural University, Taigu, Shanxi Province 030801, China
  • Received:2024-01-27 Revised:2024-03-14 Published:2024-09-07

毛建草组织培养再生体系的建立

岳康杰, 亢红伟, 刘慧欣, 王佳丽, 田旭平   

  1. 山西农业大学林学院, 山西 太谷 030801
  • 通讯作者: 田旭平,E-mail:txp8638@sina.com
  • 作者简介:岳康杰(1998-),男,汉族,山西阳泉人,硕士研究生,主要从事园林植物育种的研究,E-mail:ykj74321@163.com
  • 基金资助:
    山西农业大学科技创新基金(2020BQ37)资助

Abstract: Using leaves of Dracocephalum rupestre as explants,in this study we explored the effects of different sterialization methods and varying combinations of plant growth hormones on the initiation of callus,development of adventitious buds,cell proliferation,and root formation on the leaf explants. The results indicated that the most effective sterialization approach for the leaves involved a 30-second immersion in 75% ethanol,followed by 4-minute treatment with 0.1% HgCl2,which resulting in a remarkably low contamination rate of 11.11%. The optimal culture medium for callus initiation consisted of Murashige and Skoog (MS)+1.0 mg·L-16-Benzylaminopurine (6-BA)+0.1 mg·L-1Indole acetic acid (IAA)+1.0 mg·L-12,4-Dichlorophenoxyacetic acid (2,4-D),achieving an induction rate of 84.51%. For callus differentiation,the best medium was MS+3 mg·L-16-BA+0.50 mg·L-1Thidiazuron (TDZ)+0.50 mg·L-1IAA,with an adventitious bud differentiation rate of 66.37%. The medium promoting shoot proliferation was MS+2.0 mg·L-16-BA+0.05 mg·L-1NAA,resulting in a proliferation rate of 83.57%. The suitable rooting medium was 1/2MS+0.1 mg·L-1Naphthylacetic acid (NAA)+0.1 mg·L-1Beta-indolebutyric acid (IBA),with a rooting rate of 86.97%. The study established an in vitro culture and regeneration system for the leaf explants of Dracocephalum rupestre,providing a theoretical and technical support for the conservation of its germplasm resources and tissue culture propagation.

Key words: Callus, Adventitious buds, Proliferation, Rooting

摘要: 本研究以毛建草(Dracocephalum rupestre Hance)叶片为外植体,研究不同消毒处理和不同植物生长激素配比对叶片愈伤组织诱导、不定芽分化、增殖、生根的影响。结果表明:叶片最佳的消毒方式为75%乙醇浸泡30 s后,用0.1% HgCl2消毒4 min,污染率最低为11.11%;诱导愈伤组织的最佳培养基为MS(Murashige and Skoog)基本培养基+1.0 mg·L-16-苄氨基腺嘌呤(6-Benzylaminopurine,6-BA)+0.1 mg·L-1吲哚乙酸(Indole acetic acid,IAA)+1.0 mg·L-12,4-二氯苯氧乙酸(2,4-Dichlorophenoxyacetic acid,2,4-D),诱导率达84.51%;愈伤组织分化的最佳培养基为MS+3 mg·L-16-BA+0.50 mg·L-1噻苯隆(Thidiazuron,TDZ)+0.50 mg·L-1IAA,不定芽分化率为66.37%;不定芽增殖最佳培养基为MS+2.0 mg·L-16-BA +0.05 mg·L-1萘乙酸(Naphthylacetic acid,NAA),增殖率为83.57%;适宜生根的培养基为1/2MS基本培养基(1/2MS)+0.1 mg·L-1NAA+0.1 mg·L-1吲哚丁酸(Beta-indolebutyric acid,IBA),生根率为86.97%。本研究建立了毛建草叶片离体培养与再生体系,为其种质资源的保存和组培快繁提供理论与技术支持。

关键词: 愈伤, 不定芽, 增殖, 生根

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