中国野生葡萄叶绿体分离及叶绿体DNA提取的研究
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

现代农业产业技术体系建设专项资金(CARS30yz1);


An Optimized Chloroplast Isolation and Chloroplast DNA Extraction Protocol for Chinese Wild Grapes
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    以中国野生刺葡萄、山葡萄、桑叶葡萄和东南葡萄的成熟叶片为材料,比较柱式植物叶绿体DNAout试剂盒和改良的高盐低pH法分离叶绿体及提取cpDNA效果。结果显示:(1)2种方法均分离得到了中国野生葡萄的叶绿体,但与柱式植物叶绿体DNAout试剂盒相比,改良的高盐低pH法得到的叶绿体浓度高、杂质少,更适合中国野生葡萄叶绿体分离。(2)柱式植物叶绿体DNAout试剂盒提取的cpDNA,其OD260/OD280在1.28~1.36之间,质量浓度为4.2~7.8 ng·μL-1,质量低,不能满足后续测序要求;而改良的高盐低pH法提取的cpDNA,其OD260/OD280值在1.84~1.90之间,质量浓度为2 514.4~4 133.7 ng·μL-1,质量高,纯度好,能满足后续测序要求。研究表明,改良的高盐低pH法能简便、快速获得中国野生葡萄完整叶绿体及高质量cpDNA,并且提取的cpDNA可满足后续测序要求,为葡萄属植物叶绿体基因组的深入研究奠定了基础。

    Abstract:

    Mature leaves collected from Vitis davidii, V. amurensis, V. heyneana and V. chunganensis were used for chloroplast isolation and cpDNA extraction in this study. The two methods were the column plant chloroplast DNAout and modified highsalt lowpH method, and the results were compared with each other. (1) Both methods had separated the chloroplast of Chinese wild grapes, but the modified highsalt lowpH method obtained higher concentration and less impurity of chloroplast than that of column plant chloroplast DNAout. So the modified highsalt lowpH method was more suitable for chloroplast isolation. (2) The value of OD260/OD280 of cpDNA extracted by the column plant chloroplast DNAout was between 1.28 and 1.36, and the concentration was between 4.2 ng·μL-1 and 7.8 ng·μL-1, which did not meet the demand of subsequent chloroplast genome sequencing. In contrast, the value of OD260/OD280 of cpDNA extracted by the modified highsalt lowpH method was between 1.84 and 1.90 and the concentration was between 2 514.4 ng·μL-1 and 4 133.7 ng·μL-1, so the cpDNA extracted in this way was extremely highquality and pure. As a result, the cpDNA extracted by the modified highsalt lowpH method meet the demand of subsequent chloroplast genome sequencing. As a conclusion, the modified highsalt lowpH method isolated intact chloroplast and extract highquality cpDNA of Chinese wild grapes simply and quickly. And the cpDNA meet the demand of subsequent chloroplast genome sequencing. It was also a critical step to make further research of chloroplast genomes of Vitis L.

    参考文献
    相似文献
    引证文献
引用本文

谢海坤,焦 健,樊秀彩,等.中国野生葡萄叶绿体分离及叶绿体DNA提取的研究[J].西北植物学报,2016,36(7):1464-1469

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: 2016-08-16
  • 出版日期:

微信公众号二维码

手机版网站二维码