植物学报 ›› 2020, Vol. 55 ›› Issue (5): 588-595.DOI: 10.11983/CBB20019

• 技术方法 • 上一篇    下一篇

早花百子莲叶片器官发生和胚胎发生再生体系的建立

岳建华1,3, 董艳2, 王小画1, 孙佩霞1, 王思颖1, 张新年1, 张琰1,3,*()   

  1. 1信阳农林学院园艺学院, 信阳 464100
    2信阳农林学院林学院, 信阳 464100
    3信阳市大别山区园艺植物遗传改良重点实验室, 信阳 464100
  • 收稿日期:2020-02-10 接受日期:2020-05-08 出版日期:2020-09-01 发布日期:2020-09-03
  • 通讯作者: 张琰
  • 作者简介:E-mail: zhy8010878@163.com
  • 基金资助:
    河南省科技攻关计划(172102110263);河南省基础与前沿研究计划(162300410181);信阳农林学院青年教师科研基金(201701014);信阳农林学院园艺专业综合改革试点建设(ZYZHGG201803)

A Regeneration System for Organogenesis and Somatic Embryogenesis Using Leaves of Agapanthus praecox as Explants

Jianhua Yue1,3, Yan Dong2, Xiaohua Wang1, Peixia Sun1, Siying Wang1, Xinnian Zhang1, Yan Zhang1,3,*()   

  1. 1College of Horticulture, Xinyang Agriculture and Forestry University, Xinyang 464100, China
    2College of Forestry, Xinyang Agriculture and Forestry University, Xinyang 464100, China
    3Key Laboratory of Horticultural Plants Genetic Improvement of Dabie Mountain, Xinyang City, Xinyang 464100, China
  • Received:2020-02-10 Accepted:2020-05-08 Online:2020-09-01 Published:2020-09-03
  • Contact: Yan Zhang

摘要: 以早花百子莲(Agapanthus praecox)叶片为外植体, 建立了器官发生和胚胎发生离体再生体系, 并对移栽驯化基质进行了初步筛选。结果表明, 毒莠定(PIC)对叶片愈伤组织诱导效果良好, 最适培养基为MS+2.0 mg·L -1 PIC; 叶片组织分生能力决定愈伤组织诱导效果, 1-2片新叶基部愈伤组织诱导率可达85.71%, 叶片分生区0-0.5 cm愈伤组织诱导率为66.48%, 叶片横切面中部诱导效果优于边缘。不定芽诱导最适培养基为MS+1.5 mg·L -1 PIC+0.3 mg·L -1 6-BA, 诱导率达80.27%。体细胞胚诱导培养基为MS, 0.05 mg·L -1多效唑或1.0 mg·L -1 ABA均对体胚诱导具有显著促进作用。1.0 mg·L -1 6-BA对幼苗增殖有利, 器官发生和胚胎发生途径幼苗增殖系数分别为2.23和2.93。草炭:珍珠岩:蛭石=1:1:1 (v/v/v)为早花百子莲移栽驯化的最佳基质, 成活率达100%。该研究建立了早花百子莲叶片外植体再生体系, 丰富了百子莲快繁技术体系, 可为其它单子叶植物离体再生体系建立提供参考。

关键词: 早花百子莲, 分生能力, 器官发生, 体细胞胚胎发生, 毒莠定

Abstract: A regeneration system for organogenesis and somatic embryogenesis in vitro was established by using leaves of Agapanthus praecox as explants, and different cultivation media for transplanting were selected for the best effect. The results showed that picloram (PIC) was effective in callus induction of leaves, and the optimal medium was MS+2.0 mg·L -1 PIC. The callus induction rate was determined by the meristematic activity of leaf segments. The callus induction rate of the basal tissues on the 1 st-2 nd euphyll was 85.71%, and the callus induction rate was 66.48% in meristematic zone of 0-0.5 cm of the same leaf. The results also showed that the callus induction efficiency was higher in the middle of leaf transection compared with that at the edge. The optimal medium for adventitious bud induction was MS+1.5 mg·L -1 PIC+0.3 mg·L -1 6-BA, and the induction rate was 80.27%. The basic MS medium was suitable for somatic embryo induction, but the induction rate would be significantly increased if 0.05 mg·L -1 paclobutrazol and 1.0 mg·L -1abscisic acid were added. Plantlets proliferation was promoted by 1.0 mg·L -1 6-BA, and the proliferation coefficients of organogenesis and somatic embryogenesis pathway were 2.23 and 2.93, respectively. The combination of peat:perlite: vermiculite=1:1:1 (v/v/v) was proved the suitable substrate for transplanting and acclimatization of plantlets, with a survival rate of 100%. This regeneration system provides a rapid and efficient propagation technology for A. praecox, and also provides a reference for the regeneration of monocotyledon explants in vitro.

Key words: Agapanthus praecox, meristematic activity, organogenesis, somatic embryogenesis, picloram