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园艺学报 ›› 2022, Vol. 49 ›› Issue (1): 166-174.doi: 10.16420/j.issn.0513-353x.2020-0243

• 研究报告 • 上一篇    下一篇

‘凤丹’牡丹幼胚愈伤组织诱导的优化和再生体系的建立

刘蓉, 慈惠婷, 任秀霞, 高洁, 王顺利(), 张秀新()   

  1. 中国农业科学研究院蔬菜花卉研究所,国家花卉改良中心,中国农业科学院牡丹研究中心,农业部园艺作物生物学与种质创制重点实验室,北京 100081
  • 收稿日期:2021-10-20 修回日期:2021-11-18 出版日期:2022-01-25 发布日期:2022-01-24
  • 基金资助:
    国家重点研发计划项目(2019YFD1001500);国家现代农业产业技术体系建设专项资金项目(CARS-21);中央级公益性科研院所基本科研业务费专项(IVF-BRF2021018);中国农业科学院科技创新工程项目(CAAS-ASTIP-IVFCAAS)

Optimization of Callus Induction from Immature Embryo and Establishment Regeneration System of Paeonia ostii‘Fengdan’

LIU Rong, CI Huiting, REN Xiuxia, GAO Jie, WANG Shunli(), ZHANG Xiuxin()   

  1. Key Laboratory of Biology and Genetic Improvement of Horticultural Crops,Ministry of Agriculture,P. R. China,Research Center of Peony,Chinese Academy of Agricultural Sciences,National Center for Flower Improvement,Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081,China
  • Received:2021-10-20 Revised:2021-11-18 Online:2022-01-25 Published:2022-01-24

摘要:

以‘凤丹’牡丹的幼胚为材料,从取样时期、切割接种方式、基本培养基和植物生长调节剂选择等方面进行再生体系优化。结果表明:授粉后60 d为‘凤丹’牡丹幼胚诱导愈伤的最佳取材时期;完全破碎切割成幼胚薄片比完整胚、1/2胚和1/4胚更容易获得高质量的愈伤组织,且褐化率较低;MS基本培养基添加0.162 μmol · L-1 H2O2可提高幼胚愈伤组织诱导率,但会增加褐化率,而在WPM基本培养基中添加不同浓度的H2O2,对愈伤诱导率和褐化率的影响不大;WPM基本培养基比MS更适于‘凤丹’牡丹幼胚再生;WPM + 2.5 mg · L-1 2,4-D + 0.1 mg · L-1 6-BA是诱导愈伤的最佳植物生长调节剂组合,诱导率可达93.33%;不定芽和不定根诱导的最佳组合分别为WPM + 30 g · L-1蔗糖 + 3 g · L-1植物凝胶 + 0.5 mg · L-1 6-BA + 0.5 mg · L-1 GA3和1/2 WPM + 30 g · L-1 蔗糖 + 2.5 g · L-1植物凝胶 + 2 mg · L-1 IBA + 1 mg · L-1 IAA。

关键词: 牡丹, 幼胚, 愈伤诱导, 褐化, 离体, 再生

Abstract:

In order to improve callus induction quality,and establish highly efficient regeneration system of Paeonia ostii‘Fengdan’,the immature embryo collecting time,technical treatments of immature embryos,basic medium selection,and different concentrations and combinations of plant growth regulators and H2O2 were optimized. The results showed that the best collection time for immature embryos callus induction was 60 days after pollination. Compared with four technical treatments of immature embryos,thoroughly divided embryos(slices)were produced better callus with low callus browning rate than that with whole embryos,halved embryos,and broken into quarters embryos. MS basic medium with 0.162 μmol · L-1 H2O2 can improve the callus induction rate of immature embryo,and it can also increases the callus browning rate. Different concentration of H2O2 adding in WPM basic medium could not be improved the callus induction rate. WPM basic medium is better than MS basic medium for immature embryo callus induction. The combination of WPM + 2.5 mg · L-1 2,4-D + 0.1 mg · L-1 6-BA can obtain higher callus induction rate(93.33% ± 3.53%)with lower callus browning rate(25.33% ± 4.81%). The best hormone combinations for adventitious bud and root induction were WPM + 0.5 mg · L-1 GA3 + 0.5 mg · L-1 6-BA,1/2 WPM + 2.0 mg · L-1 IBA + 1.0 mg · L-1 IAA,respectively.

Key words: tree peony, immature embryo, callus induction, callus browning, in vitro, regeneration

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