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植物研究 ›› 2018, Vol. 38 ›› Issue (1): 56-63.doi: 10.7525/j.issn.1673-5102.2018.01.007

• 研究报告 • 上一篇    下一篇

芍药愈伤组织中体细胞胚发育过程的组织细胞学观察

魏冬霞, 张滕, 郑严仪, 于晓南   

  1. 北京林业大学园林学院/国家花卉工程技术研究中心, 北京 100083
  • 收稿日期:2017-05-17 出版日期:2018-01-15 发布日期:2018-01-06
  • 通讯作者: 于晓南,E-mail:yuxiaonan626@126.com E-mail:yuxiaonan626@126.com
  • 作者简介:魏冬霞(1991—),女,硕士研究生,主要从事芍药组织培养方面的研究.
  • 基金资助:
    北京市教育委员会科学研究与研究生培养共建项目资助(BLCXY201629);国家自然科学基金(31400591)

Histocytology Observation on the Somatic Embryogenesis in Herbaceous Peony Callus

WEI Dong-Xia, ZHANG Teng, ZHENG Yan-Yi, YU Xiao-Nan   

  1. College of Landscape Architecture, Beijing Forestry University, Beijing 100083
  • Received:2017-05-17 Online:2018-01-15 Published:2018-01-06
  • Supported by:
    Graduate Training and Development Program of Beijing Municipal Commission of Education(BLCXY201629);National Natural Science Foundation of China(31400591)

摘要: 以芍药(Paeonia lactiflora Pall.)3个芍药品种的茎段、叶片、叶柄为外植体,诱导体细胞胚发生,并采用石蜡切片法对该发育过程进行组织细胞学观察。结果表明:‘Going Bananas’的茎段愈伤诱导率达100%,增殖率在4.0以上,表现最佳;非胚性愈伤组织最佳诱导、增殖培养基为WPM+IAA 1.0mg·L-1+6-BA 1.0mg·L-1+NAA 0.5mg·L-1+TDZ 0.5mg·L-1+CH 0.625g·L-1;愈伤组织转入到1/2 MS(Ca2+加倍)+2,4-D 2.0mg·L-1+ABA 0.5mg·L-1或ZT 1.0mg·L-1的培养基中,连续暗培养90后得到胚性愈伤;之后转入到成熟培养基1/2MS(Ca2+加倍)+6-BA 1.0mg·L-1+NAA 0.2mg·L-1中,见光培养60d,逐渐发育至球形胚和心形胚。在石蜡切片观察中,芍药的体细胞胚起源方式包括外起源和内起源两个途径。在外起源方式中包括单个表层细胞的外起源和多个表层下细胞的共同起源。3种方式的区别主要在于起始的位置和起始细胞数量,后期均形成原胚结构。胚性细胞的分裂方式为对称分裂,未发现不对称分裂细胞的存在。

关键词: 芍药, 体细胞胚, 起源方式, 组织细胞学

Abstract: The stem segments, leaves and petioles of three herbaceous peony was used as explants for somatic embryogenesis. Paraffin method was used for morphological and histological observation of somatic embryogeneris. The main conclusions were as follows: callus induction rate from stems of ‘Going Bananas’ was 100%. Proliferation rate of the callus was over 4.0. The best induction and proliferation medium of inducting callus was WPM+IAA 1.0 mg·L-1+6-BA 1.0 mg·L-1+NAA 0.5 mg·L-1+TDZ 0.5 mg·L-1+CH 0.625 g·L-1. Embryogenic callus generated after transferred the callus to 1/2 MS(Ca2+ doubled)+2,4-D 2.0 mg·L-1+ABA 0.5 mg·L-1 or ZT 1.0 mg·L-1 medium of a constantly dark cultivation of 90 d. Globular embryos and heart embryos were generated after transferred the embryogenic callus to 1/2MS(Ca2+ doubled)+6-BA1.0 mg·L-1+NAA 0.2 mg·L-1 medium for a constantly illumination cultivation of 60 d. There are exogenous and endogenous origins of somatic system. Exogenous includes origins from individual surface cells and origins from multiple subcritical cells. The difference of three ways lies mainly in starting position and starting cells number, they will all generated proembryo later. The splitting of embryogenic cells was symmetrical and the presence of asymmetric splitting cells was not found.

Key words: herbaceous peony, somatic embryo, origin, histocytology

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