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中国农学通报

中国农学通报 ›› 2016, Vol. 32 ›› Issue (34): 119-122.doi: 10.11924/j.issn.1000-6850.casb16080126

所属专题： 园艺

• 生物技术科学 • 上一篇    下一篇

甜菜SCoT-PCR反应体系的建立及优化

刘乃新1,2,3,郭晨晨1,3,孙燕红1,3,余虹莹1,3,马龙彪1,2,3,吴则东1,2,3   

  1. (1黑龙江省普通高校甜菜遗传育种重点实验室/黑龙江大学,哈尔滨 150080；2中国农业科学院甜菜研究所/黑龙江大学农作物研究院,哈尔滨 150080；3中国农业科学院北方糖料作物资源与利用重点开放实验室,哈尔滨 150080）
  • 收稿日期:2016-08-26 修回日期:2016-10-17 接受日期:2016-10-25 出版日期:2016-12-15 发布日期:2016-12-15
  • 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建设项目“东北区育种”(CARS-210104)。

Establishment and Optimization of SCoT-PCR System in Sugarbeet

Liu Naixin1,2,3, Guo Chenchen1,3, Sun Yanhong1,3, Yu Hongying1,3, Ma Longbiao1,2,3, Wu Zedong1,2,3   

  1. (1Key Laboratory of Sugar Beet Genetic Breeding/Heilongjiang University, Harbin 150080; 2Sugar Beet Research Institute of Chinese Academy of Agricultural Sciences/Crop Academy of Heilongjiang University, Harbin 150080; 3Key Laboratory of North Sugar Crop Resource and Utilization, Chinese Academy of Agricultural Sciences, Harbin 150080)
  • Received:2016-08-26 Revised:2016-10-17 Accepted:2016-10-25 Online:2016-12-15 Published:2016-12-15

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摘要： 旨在利用SCoT分子标记技术为甜菜指纹图谱构建、遗传多样性分析以及其他分子标记辅助育种提供技术支持。本研究利用单因素实验优化了甜菜SCoT-PCR反应体系。结果表明，在20 μL反应体系中，甜菜SCoT-PCR最优反应体系包含2.0 μL的10×PCR buffer（含Mg2 ）、10 ng的DNA、0.5 U的Taq DNA聚合酶、10 μmol/L的引物2 μL以及0.1 μL的dNTPs (2.5 mmol/L each)。利用优化后的程序对12份糖甜菜品种进行扩增，结果表明，该体系扩增结果稳定、条带清晰，可用于甜菜品种指纹图谱的构建以及其他分子生物学领域的研究。

关键词: 不同生育时期, 不同生育时期, 水分胁迫, 花生, 生长发育, 产量

Abstract: The objective of the study is to provide technical support for fingerprint construction, genetic diversity analysis and other molecular marker assisted breeding of sugarbeet, with SCoT molecular markers. This sugarbeet SCoT-PCR reaction system was optimized by using the single factor experiment. The results showed that the SCoT-PCR optimal reaction system contained 2.0 μL of 10×PCR buffer (including Mg2 ), 10 ng of DNA template , 0.5 U of Taq polymerase, 2 μL of 10 μmol/L SCoT primers and 0.1μL of dNTPs (2.5 mmol/L each) in the 20 μL reaction system. Then, 12 varieties of sugarbeets were amplified using the optimized reaction system. The results demonstrated that this system was stable and had clear amplified band, so it could be used for constructing fingerprint of sugarbeet and other molecular biology research.