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中国农学通报

中国农学通报 ›› 2017, Vol. 33 ›› Issue (1): 137-141.doi: 10.11924/j.issn.1000-6850.casb16100018

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

新牧4号紫花苜蓿原生质体游离和培养条件的研究

陈爱萍,刘慧敏,李瑶   

  1. 新疆草地资源与生态实验室 /新疆农业大学草业与环境科学学院,新疆草地资源与生态实验室 /新疆农业大学草业与环境科学学院,新疆草地资源与生态实验室 /新疆农业大学草业与环境科学学院
  • 收稿日期:2016-10-08 修回日期:2016-12-21 接受日期:2016-11-26 出版日期:2017-02-04 发布日期:2017-02-04
  • 通讯作者: 陈爱萍
  • 基金资助:
    草业科学国家重点学科项目“提高苜蓿原生质体细胞分裂频率的研究”(XJCYQ-2012-02)。

Protoplasts Isolation and Culture Factors of Medicago sativa L.‘Xinmu No.4’

  • Received:2016-10-08 Revised:2016-12-21 Accepted:2016-11-26 Online:2017-02-04 Published:2017-02-04

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摘要： 主要探讨影响苜蓿原生质体游离和培养的条件，为原生质体的培养体系的建立提供了依据。以‘新牧4号紫花苜蓿’的子叶、下胚轴和愈伤组织为材料，研究了酶液的pH值，酶解时间，酶液组合和不同培养方法对原生质体游离和培养效果的影响。结果表明：当酶液pH值为5.8时，有活力的原生质体的产量最高，同时细胞碎片少；子叶、下胚轴和愈伤组织适宜的酶解时间均为10 h；酶液组合为2%纤维素酶＋0.5%果胶酶＋0.3%或0.5%离析酶，游离出的有活力的原生质体产量较高；采用液体浅层培养和固液双层培养，均观察到原生质体发生分裂，但固液双层培养法更有利于‘新牧4号紫花苜蓿’原生质体的分裂和培养。

关键词: 甜菜, 甜菜, 可溶性糖, 蔗糖合成酶, 蔗糖磷酸合成酶

Abstract: The factor of alfalfa protoplasts isolation and culture were discussed, which could provide a basis for the establishment of protoplast culture system. Using cotyledons, hypocotyls and hypocotyl callus of Medicago sativa L.‘Xinmu No.4’as tested materials, the effects of pH value of enzyme, enzymolysis time, enzyme combination, different culture methods on protoplasts isolation and culture were studied. The results showed that when pH value of enzyme was 5.8, the yield of viable protoplast was the highest, at the same time, cell debris was fewer. The optimum enzymolysis time of cotyledons, hypocotyls and hypocotyl callus was 10 h. The enzyme concentration was 2.0% cellulase 0.5% pectinase 0.3% (or 0.5%) macerozyme, the yield of viable protoplast was relatively high. Using liquid medium and solid-liquid double layer medium, protoplast division was observed, but solid-liquid double layer culture was more favorable for protoplast division and culture of Medicago sativa L.‘Xinmu No.4’