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植物研究 ›› 2003, Vol. 23 ›› Issue (4): 429-432.

• 论文 • 上一篇    下一篇

五针松胚乳ISSR-PCR反应体系的建立

吕艳芳, 刘桂丰, 姜静, 侯英杰   

  1. 东北林业大学森林资源与环境学院, 哈尔滨 150040
  • 收稿日期:2003-06-26 出版日期:2003-12-15 发布日期:2016-06-14
  • 作者简介:吕艳芳(1977-),女,硕士,主要从事林木遗传育种研究。
  • 基金资助:
    国家"948西伯利亚红松优良种质资源引进"资助(2001-31)

THE ISSR-PCR REACTION SYSTEM'S ESTABLISHMENT ABOUT SEED'S ENDOSPERM OF FIVE NEEDLES PINE

LÜ Yan-Fang, LIU Gui-Feng, JIANG Jing, HOU Ying-Jie   

  1. Collage of Forest Resource and Environments, Northeast Forestry University, Harbin 150040
  • Received:2003-06-26 Online:2003-12-15 Published:2016-06-14

摘要: 采用改良的CTAB法提取5种五针松胚乳DNA,经检测,该方法提取的DNA纯度和含量较高,单粒种子胚乳DNA得率基本满足大量PCR扩增的需要。建立了稳定的、可重复的五针松ISSR-PCR最佳反应体系及PCR扩增程序,筛选出了扩增条带清晰、多态性丰富的16个ISSR引物,为今后利用五针松种子胚乳开展遗传图谱构建等分子生物学研究提供一个标准化程序。

关键词: 五针松, 胚乳, DNA 提取, ISSR-PCR

Abstract: DNA was extracted from the endosperm of five needles pine by improved CTAB method. The purity of DNA that extracted by this method was high, which was tested by UV spectrophotometer. The yield of single seed's endosperm DNA almost could meet the need of mass PCR amplification. The best stable reduplicative ISSR-PCR reaction systems of five needles pine and PCR amplification parameters were established. Sixteen ISSR-PCR primers that had vivid amplification band and abundant polymorphism had been screened. The work provides a standard program for using need's endosperm of pine to carry out establishment of genetic fingerprinting and other studies of molecular biology in the future.

Key words: five needles pine, seed's endosperm, DNA extracted, ISSR-PCR