草地学报

草地学报 ›› 2014, Vol. 22 ›› Issue (4): 834-839.DOI: 10.11733/j.issn.1007-0435.2014.04.024

• 研究论文 • 上一篇    下一篇

黄花苜蓿高频植株再生体系建立的研究

李娟1, 张万军1, 王涛2   

  1. 1. 中国农业大学动物科技学院草业科学系 草业科学北京市重点实验室, 北京 100193;
    2. 中国农业大学农业与生物技术国家重点实验室, 北京 100193
  • 收稿日期:2013-11-01 修回日期:2014-01-14 出版日期:2014-08-15 发布日期:2014-08-04
  • 通讯作者: 张万军
  • 作者简介:李娟（1990-），女，甘肃白银人，硕士研究生，研究方向为牧草分子育种，E-mail：lijuan546@163.com
  • 基金资助:

    中央高校基本科研业务费专项（15052001）资助

Establishing A Highly Efficient Plant Regeneration System of Medicago falcata L.

LI Juan1, ZHANG Wan-jun1, WANG Tao2   

  1. 1. Beijing Key Laboratory for Grassland Science, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China;
    2. State Key laboratory of Agriculture and biotechnology, China Agricultural University, Beijing 100193, China
  • Received:2013-11-01 Revised:2014-01-14 Online:2014-08-15 Published:2014-08-04

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摘要：

黄花苜蓿（Medicago falcataL.）具有较强的抗逆性，是苜蓿育种重要的种质资源。进行黄花苜蓿功能基因研究，有必要建立其高效的植株再生体系。以新疆野生黄花苜蓿的子叶和下胚轴为外植体，以N6大量，MS微量及铁盐，水解酪蛋白2.0g·L-1，维生素VB19.9mg·L-1，VB69.5mg·L-1，尼克酸4.5mg·L-1，琼脂8.0g·L-1和蔗糖30g·L-1等成分为基本培养基，通过比较不同激素配比对黄花苜蓿愈伤诱导率和体细胞胚发生率的影响，优化并建立了黄花苜蓿的高频再生体系。优化的培养基激素配比为：愈伤诱导和保持培养基添加2，4-D 3.0mg·L-1+KT 0.05mg·L-1；分化培养基添加KT 0.4mg·L-1，蔗糖浓度调整为20g·L-1；生根培养基为1/2MS+蔗糖15g·L-1+琼脂8.0g·L-1。在优化培养流程下，新疆野生黄花苜蓿子叶愈伤诱导率可达92%，下胚轴可达95%，体细胞胚发生率可达54%，体细胞胚萌发成苗率为76%。再生周期约为70～80d。

关键词: 黄花苜蓿, 再生体系, 愈伤组织, 体细胞胚

Abstract:

Medicago falcata is an important germplasm resource for alfalfa breeding because of its strong stress resistance.A highly efficient plant regeneration system is necessary to study the functional genes of M.falcata.The cotyledons and hypocotyls of wild M.falcata from Xinjiang are used as explants to investigate the effects of hormones ratio on callus induction rate and somatic embryos induction rate. A highly efficient plant regeneration system of M. falcata is established in this study.The basic medium includes N6 macronutrients+MS micronutrients+MS iron+VB1 9.9 mg·L-1+VB6 9.5 mg·L-1+Nicotinic acid 4.5 mg·L-1+CH 2.0 g·L-1+agar 8.0 g·L-1， and sucrose 30 g·L-1.For callus induction and maintenance， 2,4-D 3.0 mg·L-1 and KT 0.05 mg·L-1 are added. During somatic embryos induction stage， KT 0.4 mg·L-1 are added and sucrose is adjusted to 20 g·L-1. Medium for somatic embryo germination and plantlet rooting consists of 1/2 MS+sucrose 15 g·L-1+agar 8.0 g·L-1. In this system， the callus induction rate of cotyledon is more than 92%， and that of hypocotyl is more than 95%， while the induction rate of somatic embryos is 54%, and the rate of somatic embryos developing into whole plantlets is 76%.The period of regeneration is about 70~80 days.

Key words: Medicago, falcata, L., Regeneration, system, Callus, Somatic, embryo

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