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中国农学通报

中国农学通报 ›› 2011, Vol. 27 ›› Issue (31): 147-151.

所属专题： 园艺

• 林学 园艺 园林 • 上一篇    下一篇

洋葱愈伤原生质体分离和纯化研究

陈丽 梁毅 汪李平 田保华   

  • 收稿日期:2011-06-24 修回日期:2011-10-17 出版日期:2011-12-05 发布日期:2011-12-05
  • 基金资助:

    国家公益性行业科技“葱姜蒜产业发展关键技术研究与开发”;北京市常规育种财政专项“蔬菜种质资源的收集保存和评价利用”;北京农林科学院青年科研基金项目“洋葱异源细胞质雄性不育系快速转育技术及育种应用研究”

Studies on the Protoplast Isolation and Purification of Allium cepa

  • Received:2011-06-24 Revised:2011-10-17 Online:2011-12-05 Published:2011-12-05

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摘要：

建立高效原生质体分离和植株再生体系是进行体细胞杂交的基础性工作。以洋葱多次继代的愈伤组织为研究试材，对原生质体分离纯化技术进行研究。结果表明：2.0%纤维素酶Onozuka R-10+0.1%果胶酶Y-23+0.5%离析酶 R-10+0.60 mol/L甘露醇+CPW+0.1% MES的酶液，酶解6 h，洋葱原生质体分离效果最好。纯化时，适当降低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600 r/min离心5 min效果为好。

关键词: 猪高热病, 猪高热病, 中药, 非特异性免疫力, 病毒性疾病

Abstract:

The basic work for the system establishment about protoplast culture and somatic hybrids is to formulate efficient protoplast isolation and protoplast-culturing system. The conditions of isolation and purification of protoplast from somatic callus of Allium cepa were studied. The results showed that the optimal enzyme solution for protoplast isolation was 2.0% cellulase Onozuka R-10+ 0.1% pectolgase Y-23+0.5% macerozyme R-10+0.60 mol/L mannitol +0.1% MES and CPW solution. The enzyme mixture was for 5 h at (25±2)℃. High yield and viability protoplasts were obtained when purified by centrifugation at 600 r/min for 5 min and the callus for 12 days.