草地学报 ›› 2008, Vol. 16 ›› Issue (1): 50-53.DOI: 10.11733/j.issn.1007-0435.2008.01.010

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酶解对苜蓿子叶原生质体分离效果的影响

赵红娟, 张博, 陈爱萍, 李培英, 肖燕   

  1. 新疆农业大学草业工程学院, 新疆草地资源与生态自治区重点实验室, 新疆乌鲁木齐, 830052
  • 收稿日期:2006-12-22 修回日期:2007-11-19 出版日期:2008-02-15 发布日期:2008-02-15
  • 通讯作者: 张博, E-mail:xjauzb@126.com
  • 作者简介:赵红娟(1979- ),女,宁夏人,硕士研究生,研究方向为牧草生物技术育种,E-mail:xjzhaohj@yahoo.com.cn
  • 基金资助:

    新疆草地资源与生态自治区重点实验室开放课题(XJDX0201-2004-04);新疆自治区科技厅高新技术项目(200511102)

Effects of Different Enzymolysis Conditions on the Cotyledons Protoplast Isolation of Alfalfa

ZHAO Hong-juan, ZHANG Bo, CHEN Ai-ping, LI Pei-ying, XIAO Yan   

  1. Xinjiang Key Laboratory of Grassland Resources and Ecology, College of Pratacultural Engineering, Xinjiang Agricultural University, Urumqi, Xinjiang Uygur Autonomous Region, 830052, China
  • Received:2006-12-22 Revised:2007-11-19 Online:2008-02-15 Published:2008-02-15

摘要: 以新牧1号杂花苜蓿(Medicago varia Martyn cv. Xinmu No.1)无菌苗子叶为材料,研究酶液组合、酶解时间、酶液渗透压及酶液pH值对其原生质体的分离效果。结果表明:获得有活力原生质体最佳的酶液组合、酶解时间、酶液渗透压和酶液pH值,分别为2%纤维素酶+0.5%果胶酶+0.3%离析酶、10h、0.55mol/L和5.6。

关键词: 苜蓿, 原生质体, 酶解条件

Abstract: The Effects of different enzyme combination,mannitol concentration,enzymolysis time,and pH on protoplast isolation were investigated using the aseptic seedling cotyledons of Medicago varia Martyn cv.Xinmu No.1 as tested materials.The results show that the optimum conditions for high yield of viable protoplast were as follows: enzyme combination containing 2% cellulose onozuka R-10+0.5% pectinase+0.3% macerozyme R-10,mannitol concentration at 0.55 mol/L,enzymolysis time of 10 h,and pH=5.6.

Key words: Medicago varia Martyn, Protoplast, Enzymolysis conditions

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