植物学报 ›› 2011, Vol. 46 ›› Issue (2): 170-178.DOI: 10.3724/SP.J.1259.2011.00170

• 研究报告 • 上一篇    下一篇

雷竹体细胞胚诱导的初步研究

裴海燕1, 林新春1, 方伟1, 黄丽春2*   

  1. 1浙江农林大学亚热带森林培育国家重点实验室培育基地, 临安 311300
    2台湾“中央研究院”植物研究所, 台北 115
  • 收稿日期:2010-03-02 修回日期:2010-05-28 出版日期:2011-03-01 发布日期:2011-05-09
  • 通讯作者: 黄丽春
  • 基金资助:

    省级自然科学基金

A Preliminary Study of Somatic Embryogenesis of Phyllostachys violascens In Vitro

Haiyan Pei1, Xinchun Lin1, Wei Fang1, Lichun Huang2*   

  1. 1The Nurturing Station for the State Key Laboratory of Subtropical Silviculture, Zhejiang Agriculture and Forestry University,Lin’an 311300, China;

    2Institute of Botany, “Academia Sinica”, Taibei 115, China
  • Received:2010-03-02 Revised:2010-05-28 Online:2011-03-01 Published:2011-05-09
  • Contact: Lichun Huang

摘要: 以雷竹(Phyllostachys violascens)种胚为外植体, 脱分化产生愈伤组织, 愈伤组织诱导产生体细胞胚, 并发育成苗。实验结果表明: 愈伤组织诱导体细胞胚基本培养基为MS无机盐+20 g·L–1葡萄糖(glucose)+10 mg·L–1腺嘌呤(adenine sulfate)+ 0.5 g·L–1麦芽抽提物(malt extract)+0.1 mg·L–1 6-BA+0.01 mg·L–1氨氯吡啶酸(picloram)+10 g·L–1type A agar; 将基本培养基中的氨氯吡啶酸浓度升高至0.1 mg·L–1即为愈伤组织诱导的最佳培养基, 早期子叶胚是愈伤组织诱导的最佳胚体; 愈伤组织在添加0.001 mg·L–1TDZ和0.3 mg·L–1ABA的愈伤组织最佳培养基上光照培养4周后, 去除其中的ABA, 并添加0.1 mg·L–1NAA继续培养1个月, 体细胞胚数量最多可达87.43%, 该培养基是体细胞胚发生的最佳培养基; 将上述体细胞胚发生培养基中的6-BA浓度升高至1 mg·L–1, 继代培养2个月后有小苗出现。组织学观察显示, 体细胞胚细胞核大、质浓且多数呈球形原胚状。

Abstract: Phyllostachys violascens zygotic embryos served as explants in inducing somatic embryo and subsequent plant regeneration. Murashige and Skoog salts with 20 g·L–1 glucose, 10 mg·L–1 adenine sulfate, 0.5 g·L–1 malt extract, 0.1 mg·L–1 6-BA, 0.01 mg·L–1 4-amino-3,5,6-trichloropicolinic acid (picloram) and 10 g·L–1 type A agar was selected as the basal medium for inducing somatic embryo. Early cotyledon embryos were used as explants for inducing compact callus formation in basal medium with increased picloram (0.1 mg·L–1). This medium was the best for inducing embryogenic callus. Somatic embryogenesis reached 87.4% when embryogenic callus was transferred to the medium and supplemented with 0.001 mg·L–1 thidiazuron and 0.3 mg·L–1 abscisic acid for 4 weeks, then abscisic acid removed and 0.1 mg·L–1 N-acetyl-aspartate added for 1 month. Small plantlets were finally observed when 6-BA was increased to 1 mg·L–1 after 2 months in culture. Histological examination of somatic embryo illustrated large nuclei and dense cytoplasm cells, paired with globular proembryos.