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中国农学通报

中国农学通报 ›› 2019, Vol. 35 ›› Issue (35): 59-66.doi: 10.11924/j.issn.1000-6850.casb18100074

所属专题： 生物技术； 油料作物； 园艺

• 林学 园艺 园林 • 上一篇    下一篇

油菜不同类型外植体组织培养及再生研究

万丽丽1, 王转茸2, 辛强3, 洪登峰3, 杨光圣3   

  1. 1.华中农业大学作物遗传改良国家重点实验室B310;2.武汉市农业科学院作物研究所;3.华中农业大学
  • 收稿日期:2018-10-22 修回日期:2019-11-18 接受日期:2019-03-01 出版日期:2019-12-16 发布日期:2019-12-16
  • 通讯作者: 万丽丽
  • 基金资助:
    武汉市农科院创新项目“利用基因编辑系统创制健康优质及适宜机械化收获的油菜新品种”(CX201815)。

Tissue Culture and Regeneration Study of Various Explants of Brassica napus

  • Received:2018-10-22 Revised:2019-11-18 Accepted:2019-03-01 Online:2019-12-16 Published:2019-12-16

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摘要： 为建立高效的组织培养体系，以甘蓝型油菜杂交品种恢复系627R、621R和616R材料田间种植植株的侧芽、花托和无菌种子实生苗下胚轴为外植体，探索不同苗龄、预培养时间、预培养基、愈伤分化培养基、诱导出芽培养基以及成苗壮苗培养基中激素配比对芽再生、成苗植株生长势的影响。结果表明：无菌苗快速繁殖体系中，发芽6 天的无菌苗下胚轴或者子叶在预培养（MS+ 2.0 mg/L 2,4-D+ 1.0 mg/L 6-BA+ 30 g/L 蔗糖+ 8 g/L 琼脂，pH 5.85）3 天后转移到分化培养基MS+ 3 mg/L 6-BA+ 1.0 mg/L NAA+5 mg/L AgNO3+ 30 g/L 蔗糖+8 g/L 琼脂(pH 5.85)，或者MS+ 3 mg/L 6-BA+ 1.0 mg/L IAA+ 5 mg/LAgNO3+ 30 g/L 蔗糖+8 g/L 琼脂(pH 5.85)生长，可以获得较高的芽再生频率，对于田间生长到抽薹开花期植株取样的外植体，腋芽的出芽频率高于花托培养的出芽频率，但是这2 类外植体在分化培养基（MS+ 10 mg/L 6-BA+ 1 mg/L NAA+ 30 g/L 蔗糖+8 g/L 琼脂，pH 5.85）生长30 天后都有成苗的潜力，上 述外植体经过组织培养出芽后转移到添加矮壮素的培养基（MS+15 mg/L CCC+15~20 g/L 蔗糖+8 g/L 琼脂，pH 5.85）上继续生长30 天，能够得到根系发达、生长势强的植株。甘蓝型油菜优良恢复系建立的组织培养体系能够快速获得生长势优的油菜植株，加速油菜良种的繁殖与评价。

关键词: 施氮量, 施氮量, 施氮时期, 糯高粱, 产量和品质

Abstract: The lateral bud, flower bud receptacle, hypocotyl and cotyledon derived from sterile seedlings of elite Brassica napus restorer line 627R, 621R and 616R were used as explants for tissue culture. The research explored the effects of different seedling ages, pre- culture time, different hormone ratios of pre- culture medium, callus differentiation medium, shoot induced medium and shoot growth medium on shoot regeneration and plant growth vigor. The high frequency of shoot regeneration resulted from hypocotyl and cotyledon derived from 6 days growth seedlings cultured in the following procedure, after 3 days in pre-culture medium (MS+ 2.0 mg/L 2,4-D+ 1.0 mg/L 6-BA+ 30 g/L sucrose+ 8 g/L agar, pH 5.85, the explants were transferred to callus differentiation medium, MS+ 3 mg/L 6-BA+ 1.0 mg/L NAA+ 5 mg/L AgNO3+ 30 g/L sucrose+ 8 g/L agar, pH 5.85, or MS+ 3 mg/L 6-BA+ 1.0 mg/L IAA+ 5 mg/L AgNO3+ 30 g/L sucrose+ 8 g/L agar, pH 5.85. The high frequency of shoot regeneration derived from lateral and buds in the flower bud development could be cultured 30 days in callus or shoot regeneration medium(MS+ 10 mg/L 6-BA+ 1 mg/L NAA+ 30 g/L sucrose+ 8 g/L agar，pH 5.85). All the plants from explants culture had abundant fine roots such as increased root hair number and length, and excellent seedling vigor after transferring to medium with 15 mg/L CCC (MS+ 15 mg/L CCC+ 15-20 g/L sucrose+ 8 g/L agar，pH 5.85) for 30 days. The tissue culture system established for different explants of Brassica napus restorer line will accelerate the reproduction and evaluation of rapeseed varieties.