植物学报 ›› 2021, Vol. 56 ›› Issue (4): 443-450.DOI: 10.11983/CBB20195

• 技术方法 • 上一篇    下一篇

芍药花药愈伤组织诱导及体细胞胚发生

李艳敏, 蒋卉, 符真珠, 张晶, 袁欣, 王慧娟, 高杰, 董晓宇, 王利民, 张和臣*()   

  1. 河南省农业科学院园艺研究所, 郑州 450002
  • 收稿日期:2020-12-01 接受日期:2021-05-27 出版日期:2021-07-01 发布日期:2021-06-30
  • 通讯作者: 张和臣
  • 作者简介:*E-mail: zhc5128@126.com
  • 基金资助:
    河南省农业科学院自主创新项目(2021ZC20);河南省科技攻关计划(192102110148);河南省农业科学院科技创新创意项目(2020CX13);河南省农业科学院优秀青年基金(2020YQ17)

Callus Induction and Somatic Embryogenesis in Anther Culture of Paeonia lactiflora

Yanmin Li, Hui Jiang, Zhenzhu Fu, Jing Zhang, Xin Yuan, Huijuan Wang, Jie Gao, Xiaoyu Dong, Limin Wang, Hechen Zhang*()   

  1. Horticultural Research Institute, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China
  • Received:2020-12-01 Accepted:2021-05-27 Online:2021-07-01 Published:2021-06-30
  • Contact: Hechen Zhang

摘要: 以芍药(Paeonia lactiflora)品种粉玉奴花药为外植体, 研究不同浓度2,4-D对愈伤组织诱导、体胚发生及植株再生的影响, 采用组织细胞学方法观察愈伤组织以及体细胞胚发育过程, 采用根尖染色体法鉴定再生植株倍性。结果表明, 芍药花药愈伤组织诱导的最适培养基为MS+1 mg·L-12,4-D+1 mg·L-1 NAA+0.1 mg·L-1 KT+30 g·L-1蔗糖+6.5 g·L-1琼脂, 愈伤组织诱导率为14.7%。转入体细胞胚诱导培养基上, 历经球形胚、心形胚、鱼雷胚和子叶胚阶段, 体胚诱导率为52.1%; 在成苗培养基中能够长出真叶并得到完整植株, 成苗率为47.1%。经根尖染色体法鉴定出单倍体和二倍体植株。该研究初步建立了通过体细胞胚间接发生途径实现植株再生的培养体系, 可为芍药属其它品种的花药培养提供借鉴, 获得的再生植株是芍药遗传学研究和育种工作的重要材料。

关键词: 芍药, 花药培养, 愈伤组织, 体细胞胚, 再生植株

Abstract: The anthers of Paeonia lactiflora cv. ‘Fenyunu’ were used as explants to study the effects of different concentrations of 2,4-D on callus induction, somatic embryogenesis and plant regeneration. The cell composition of callus and the development process of somatic embryos were observed with cytohistological method, and the ploidy of regenerated plants was identified using root tip squash method. The results showed that the suitable medium for callus induction of P. lactiflora anther was MS+1 mg·L-12,4-D+1 mg·L-1NAA+0.1 mg·L-1KT+30 g·L-1sucrose+6.5 g·L-1agar, and the callus induction rate was 14.7%. The callus was transferred to somatic embryo induction medium and underwent stages of spherical embryo, heart-shaped embryo, torpedo embryo and cotyledon embryo, and the somatic embryo induction rate was 52.1%. Genuine leaves germinated in seedling medium and complete plants were obtained, and the seedling rate was 47.1%. Haploid and diploid plants were identified using root tip squash method. The study preliminarily established a culture system to implement plant regeneration through somatic embryogenesis, which also provided reference protocol for anther culture of other varieties of Paeonia. Regenerated plants are important materials for genetic research and haploid breeding of P. lactiflora.

Key words: Paeonia lactiflora, anther culture, callus, somatic embryos, regenerated plants