中国农业科学 ›› 2012, Vol. 45 ›› Issue (14): 2913-2922.doi: 10.3864/j.issn.0578-1752.2012.14.014

• 园艺 • 上一篇    下一篇

利用基于RAPD标记的MCID法快速鉴定72个葡萄品种

 王玉娟, 张彦, 房经贵, 刘崇怀, 宋长年, 孙欣   

  1. 1.南京农业大学园艺学院/江苏省果树品种改良与种苗繁育工程中心,南京 210095
    2.中国农业科学院郑州果树研究所,郑州 450009
  • 收稿日期:2011-12-30 出版日期:2012-07-15 发布日期:2012-04-10
  • 通讯作者: 通信作者房经贵,Tel:025-84399069;E-mail:fanggg@njau.edu.cn
  • 作者简介:王玉娟,E-mail:wyj6wyj@163.com
  • 基金资助:

    农业部“948”项目(2011-421)、国家教育部新世纪优秀人才支持计划项目(NCET - 08- 0796)

Rapid Identification of 72 Grape Cultivars by Using RAPD Markers-Based MCID Method

 WANG  Yu-Juan, ZHANG  Yan, FANG  Jing-Gui, LIU  Chong-Huai, SONG  Chang-Nian, SUN  Xin   

  1. 1.南京农业大学园艺学院/江苏省果树品种改良与种苗繁育工程中心,南京 210095
    2.中国农业科学院郑州果树研究所,郑州 450009
  • Received:2011-12-30 Online:2012-07-15 Published:2012-04-10

摘要: 【目的】葡萄种质材料和品种的鉴定是葡萄种质资源研究和新品种保护的基础。本研究应用一种新的基于DNA分子标记的人工绘制品种鉴别示意图方法(manual cultivar identification diagram, MCID)对来源于不同国家和地区的72个葡萄品种进行鉴定。【方法】通过增加引物长度以及严格筛选PCR退火温度优化的RAPD技术体系,从35个长度为11个碱基的引物中筛选出6个条带清晰且多态性高的引物,进一步对72个葡萄品种进行PCR扩增获得DNA指纹,然后利用MCID法成功将将72个葡萄品种区分开来。【结果】利用这6条引物扩增的多态性谱带构建了72个葡萄品种的MCID。该葡萄MCID很好地提供对其中某些品种进行鉴定所需要的引物以及需要参考的特征性谱带。【结论】到目前为止,MCID方法是实现利用DNA分子标记鉴定果树品种能力最好的有效途径。该研究所获得的MCID具有高操作性和实用参考性,对于葡萄种质资源研究、品种保护和品种纯度早期鉴定具有重要帮助。

关键词: 葡萄, RAPD, 品种鉴定

Abstract: 【Objective】 It is a fundamental work to identify grape germplasm materials and cultivars for exploration and collecting of germplasm resources as well as plant variety protection. A new approach (manual cultivar identification diagram, MCID) by using RAPD marker was adopted to distinguish 72 cultivars of grape germplasm from different countries and areas. 【Method】 In this study, based on the optimization of RAPD through increasing the length of the RAPD primers and strict screening PCR annealing temperature, 6 effective primers were screened from 35 RAPD arbitrary 11-mer primers, and in which 6 effective primers had clear amplification products and amplified polymorphic bands, and furthermore to conduct PCR amplification. Using this strategy of MCID, all the cultivars were gradually and completely separated by a combination of the 6 primers. 【Result】A manual cultivar identification diagram (MCID) of the 72 grape cultivars was constructed using polymorphic bands from the DNA fingerprints. The MCID method could separate all the cultivars from each other, with the polymorphic bands employed for identifying the cultivars and the corresponding primers being marked in the correct position on the MCID. 【Conclusion】So far, the MCID method is the best and most effective one to realize the identification of fruit cultivars by using DNA molecular markers. The MCID of 72 grape cultivars has high operability and practicability in identifying varieties using DNA markers, which can be of much help in grape cultivars identification for protection of cultivar rights and for early identification of seedlings in the nursery industry, thus providing a theoretical basis for germplasm evaluation and cultivar identification.

Key words: grape, RAPD, cultivar identification