植物学报 ›› 2015, Vol. 50 ›› Issue (3): 378-387.DOI: 10.3724/SP.J.1259.2015.00000

• 技术方法 • 上一篇    下一篇

百合未授粉子房离体培养胚胎形成及植株再生

袁素霞, 李佳, 明军*(), 刘春, 徐雷锋, 袁迎迎   

  1. 中国农业科学院蔬菜花卉研究所, 北京 100081
  • 收稿日期:2014-05-13 接受日期:2014-08-25 出版日期:2015-05-01 发布日期:2015-04-08
  • 通讯作者: 明军
  • 作者简介:

    ? 共同第一作者

  • 基金资助:
    国家自然科学基金(No.31301804)、国家科技支撑计划(No.2012BAD01B0701)、北京市花卉重点项目(No.YLHH201000101, No.YLHH201400101)和农业部园艺作物生物学与种质创制重点实验室项目

Embryogenesis and Plant Regeneration from Unpollinated Ovary Culture of Lily

Suxia Yuan, Jia Li, Jun Ming*, Chun Liu, Leifeng Xu, Yingying Yuan   

  1. Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2014-05-13 Accepted:2014-08-25 Online:2015-05-01 Published:2015-04-08
  • Contact: Ming Jun
  • About author:

    ? These authors contributed equally to this paper

摘要:

未受精子房离体培养是诱导雌核产生单倍体的技术之一。以1个野生种和3个杂种系共7个百合(Lilium)基因型为实验材料, 探讨了基础培养基、花蕾取样时期和外源激素等因素对百合未授粉子房离体培养胚胎形成的影响。结果表明, CBM、MS和BDS三种基础培养基均可诱导百合未授粉子房胚胎形成, 但以BDS培养基诱导效果最佳; 开花前1天的花蕾较适于百合未授粉子房离体培养; 2 mg·L-1 2,4-D + 2 mg·L-1 6-BA和2 mg·L-1 2,4-D + 2 mg·L-1 KT两种外源激素配方均适用于百合未授粉子房离体培养诱导胚胎形成。在培养过程中, 大多数胚性胚珠中只含有1个胚胎, 位于珠孔端、合子端或极核处, 少数胚性胚珠中含有双胚胎。通过百合未授粉子房离体培养, 从5个基因型材料中共获得146株再生植株。采用根尖染色体计数法对其中的62株进行了倍性测定, 其中43株与母体植株染色体倍性不同。

Abstract:

Unfertilized ovary or ovule culture is an effective technology for producing homozygous materials (doubled haploids) via gynogenesis. We studied the effects of basal medium and bud developmental stage on the genogenesis in seven lily genotypes, including one wild species and six cultivars. Gynogenesis occurred in all genotypes, but BDS was the optimal basal medium as compared with CBM and MS. When ovary sections from buds collected 1 d before anthesis were cultured in BDS medium, more embryogenic ovules or responding ovules were obtained. To estimate the effect of phytohormones on lily gynogenic embryogenesis, two genotypes were tested. An amount of 2 mg·L-1 2,4-dichloro- phenoxyacetic acid (2,4-D) and 2 mg·L-1 6-benzylaminopurine (6-BA) or kinetin (KT) was suitable for embryo induction from unfertilized ovaries. During culture, embryo development was observed by the cleaning-squash technique; most ovules contained one embryo, which lay on the micropylar end, zygote end or polar nucleus position. In addition, a few ovules with two embryos were observed. In total, 146 regenerants were obtained from unpollinated ovary culture of five genotypes; for 62, the ploidy level was determined by chromosome counting. We found that 43 plantlets had a chromosome number different from maternal plants.