油菜不同类型外植体组织培养及再生研究

Tissue Culture and Regeneration Study of Various Explants of Brassica napus

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/L AgNO3+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。

The lateral bud,flower bud receptacle,hypocotyl and cotyledon derived from sterile seedlings of elite Brassica napus restorer line 627 R,621 R and 616 R 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,pH5.85,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 cultured30 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.