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中国农学通报 ›› 2010, Vol. 26 ›› Issue (4): 195-199.

所属专题: 园艺

• 林学 园艺 园林 • 上一篇    下一篇

大蒜原生质体游离和纯化的研究

杨茹,刘世琦,张自坤,刘中良,张立伟,马琳   

  • 收稿日期:2009-09-16 修回日期:2009-10-23 出版日期:2010-02-20 发布日期:2010-02-20
  • 基金资助:

    国家科技支撑项目:园艺作物基因资源挖掘与种质创新利用研究;山东省农业重大创新项目:葱姜蒜安全生产关键技术。

The protoplast isolation and purification of Allium sativum L.

  • Received:2009-09-16 Revised:2009-10-23 Online:2010-02-20 Published:2010-02-20

摘要:

以大蒜胚性悬浮细胞系为研究试材,对原生质体分离纯化技术进行了研究。结果表明:2.0%纤维素酶Onozuka R-10+0.2%果胶酶Pectolgase+0.55 mol/L甘露醇+5mM Cacl2+5mM MES的酶液,酶解时间5h,大蒜原生质体分离效果最好。纯化时,适当提高离心力的短时间离心有利于原生质体纯化过程中产量和活力的保持,以1000 r/min离心5min,效果为好。

关键词: 矿山废弃地, 矿山废弃地, 生态恢复, 土壤基质, 土壤改良

Abstract:

The conditions of isolation and purification of protoplast from somatic embryogenic cell suspension of Allium sativum L. were studied. The results showed that the optimum condition was obtained when using 2.0% Cellulase Onozuka R-10,0.2% Pectolgase,0.55 mol/L mannitol, 5mM MES for 5h. High yield and viability protoplasts were obtained when purified by centrifugation at 1000r/min for 5min.