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中国农学通报 ›› 2016, Vol. 32 ›› Issue (33): 143-146.doi: 10.11924/j.issn.1000-6850.casb16080032

所属专题: 园艺

• 生物技术科学 • 上一篇    下一篇

不同检测方法对甜菜ISSR引物多态性的影响

刘乃新,余虹莹,韦藜,马龙彪,吴则东   

  1. 黑龙江大学农作物研究院,黑龙江大学生命学院,黑龙江大学生命学院,黑龙江大学农作物研究院,黑龙江大学农作物研究院
  • 收稿日期:2016-08-05 修回日期:2016-09-25 接受日期:2016-10-11 出版日期:2016-11-25 发布日期:2016-11-25
  • 通讯作者: 吴则东
  • 基金资助:
    国家甜菜现代农业产业技术体系东北区育种岗位专家(CARS-210104)。

Effects of Different Detection Methods on ISSR Primer Polymorphism of Sugar Beet

  • Received:2016-08-05 Revised:2016-09-25 Accepted:2016-10-11 Online:2016-11-25 Published:2016-11-25

摘要: 为了探讨不同检测方法对甜菜ISSR引物扩增效果的影响以及各方法的优缺点,笔者应用非变性聚丙烯酰胺凝胶电泳和琼脂糖凝胶电泳两种检测方法对甜菜ISSR 引物扩增效果进行了检测。利用12个不同的甜菜品种,对10 个ISSR引物进行扩增,分别采用6%的非变性聚丙烯酰胺凝胶电泳、1%的琼脂糖凝胶电泳以及2%的琼脂糖凝胶电泳对ISSR-PCR的扩增产物进行分离。结果表明:6%的聚丙烯酰胺凝胶电泳无论是检测的总条带数还是多态性条带数均远高于琼脂糖凝胶电泳,且条带清晰,易于识别,而2%的琼脂糖凝胶电泳检测的条带数高于或等于1%的琼脂糖凝胶电泳,且条带更易于识别,因此,对于利用ISSR 引物进行QTL定位、指纹图谱构建以及遗传多样性分析时,推荐使用6%的非变性聚丙烯酰胺凝胶电泳,而对于种子纯度鉴定以及需要对ISSR扩增产物进行测序,则推荐使用2%的琼脂糖凝胶电泳。

关键词: 茉莉酸甲酯, 茉莉酸甲酯, ‘红地球’, 抗逆性, 高寒地区

Abstract: To investigate the effects of different detection methods on sugar beet ISSR primer polymorphism and their advantages and disadvantages, two kinds of detection methods, non-denaturing polyacrylamide gel electrophoresis and agarose gel electrophoresis were used to detect the amplification effect of sugar beet ISSR primers. 12 different varieties of sugar beet and 10 ISSR primers were used to amplify the production which were detected respectively by 6% non- denaturing polyacrylamide gel electrophoresis, 1% agarose gel electrophoresis and 2% agarose gel electrophoresis to separate the production of ISSR- PCR. The results showed that the number of polymorphic fragments or total bands detected by 6% non- denaturing polyacrylamide gel electrophoresis was more than that of agarose gel electrophoresis, and they were cleaner and more legible. However, the total bands on 2% agarose gel electrophoresis were more than or equal to that on 1% agarose gel electrophoresis. Thus, when we conduct QTL mapping, construct fingerprinting and analyze genetic diversity with ISSR primers, 6% non-denaturing polyacrylamide gel electrophoresis is recommended. While, for seed purity identification and the sequencing of ISSR amplification production, 2% agarose gel electrophoresis is more appropriate.