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植物研究 ›› 2011, Vol. 31 ›› Issue (3): 300-305.doi: 10.7525/j.issn.1673-5102.2011.03.009

• 论文 • 上一篇    下一篇

南蛇藤原生质体培养及植株再生

范小峰1;李东波1;刘灵霞1;刘秀丽1;杨颖丽2*   

  1. 1.甘肃省高校陇东生物资源保护与利用省级重点实验室,陇东学院,庆阳 745000;2.西北师范大学生命科学院,兰州 730070
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-05-20 发布日期:2011-05-20
  • 通讯作者: 杨颖丽
  • 基金资助:
     

Protoplast Culture and Plant Regeneration of Celastrus orbiculatus Thunb.

FAN Xiao-Feng;LI Dong-Bo;LIU Ling-Xia;LIU Xiu-Li;YANG Ying-Li*   

  1. 1.University Provincial Key Laboratory for Protection and Utilization of Longdong Bioresources in Gansu Province,Longdong University,Qingyang 745000;2.College of Life Science,Northwest Normal University,Lanzhou 730070
  • Received:1900-01-01 Revised:1900-01-01 Online:2011-05-20 Published:2011-05-20
  • Contact: YANG Ying-Li
  • Supported by:
     

摘要: 以4℃低温暗处理24 h的南蛇藤胚性愈伤组织为分离原生质体的原材料,用MS培养基进行液体浅层静置、固液双层以及琼脂糖包埋培养原生质体,获得再生愈伤组织并分化成苗,建立了原生质体培养体系。结果表明,低温暗处理利于高产率高质量原生质体的获得;0.5%纤维素酶+0.5%果胶酶+5 mmol·L-1 MES为酶的最佳配方;12 h为最佳酶解时间;13%为甘露醇最佳浓度;静置12 h+振荡0.5 h为最佳酶解方式;液体浅层静置培养取得了较好的原生质体培养效果;MS+6-BA 2.0 mg·L-1+IBA 0.1 mg·L-1为愈伤组织最佳分化培养基;1/2MS+NAA 0.1 mg·L-1为最佳生根培养基。

关键词: 南蛇藤, 愈伤组织, 原生质体培养, 植株再生

Abstract: Experimental raw material of protoplast was obtained from embryo callus induction of Celastrus orbiculatus Thunb. under the condition of lower temperature at 4℃ and dark treatment for 24 h. The created culture system of protoplast originated from seedlings differentiation of callus regeneration after the culture of settling the shallow-layer solution and solid-liquid double layer culture as well as agarose-embedding based on MS medium. The results showed that: the lower temperature and dark treatment was beneficial to obtain protoplast with high yield and top quality; the optimum combination for the enzyme activity would be: 0.5% Cellulase+0.5% Pectinase+5 mg·L-1 MES; the optimum time of enzymatic hydrolysis was 12 h; the optimum mannitol concentration was 13%; moreover, the optimum mode of enzymatic hydrolysis would be: standing for 12 h and oscillating for 0.5 h; meanwhile, better protoplast culture efficiency was based on the culture of settling the shallow-layer solution; the optimum differentiation medium of callus was MS+6-BA 2.0 mg·L-1+IBA 0.1 mg·L-1 and rooting medium could be 1/2 MS+NAA 0.1 mg·L-1.

Key words: Celastrus orbiculatus Thunb., callus, protoplast culture, plant regeneration

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