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植物研究 ›› 2009, Vol. 29 ›› Issue (3): 352-356.doi: 10.7525/j.issn.1673-5102.2009.03.019

• 论文 • 上一篇    下一篇

香蕉基因组SRAP反应体系的建立和优化

魏军亚1;刘德兵2;魏守兴1;谢子四1;陈业渊1*   

  1. (1.中国热带农业科学院热带作物品种资源研究所,农业部热带作物种质资源利用重点开放实验室,儋州571737) (2.海南大学园林园艺学院,儋州571737)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2009-05-20 发布日期:2009-05-20
  • 通讯作者: 陈业渊
  • 基金资助:
     

Establishment and Optimization of SRAP-PCR Reaction System in Banana Genome

WEI Jun-Ya;LIU De-Bing;WEI Shou-Xing;XIE Zi-Si;CHEN Ye-Yuan*   

  1. (1.Germplasm Research Institute of Tropical Crops of Chinese Academy of Tropical Agriculture Sciences,Key Tropical Crops Germplasm Utilization,Ministry of Agriculture P.R. of China,Danzhou571737) (2.College of Horticulture and Forestry,Hainan University,Danzhou571737)
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-05-20 Published:2009-05-20
  • Contact: CHEN Ye-Yuan
  • Supported by:
     

摘要: 为建立并优化适于香蕉(Musa spp.)SRAP分析的扩增体系,对影响香蕉SRAP反应的dNTP、Mg2+、模板DNA、引物浓度和Taq酶用量等因素进行优化。确定的优化扩增体系为Mg2+ 2.5 mmol·L-1,dNTP 250 μmol·L-1Taq酶1.0 U,引物0.5 μmol·L-1,模板DNA 20 ng,10×PCR buffer 2.5 μL,在此条件下SRAP扩增香蕉基因组DNA条带清晰,多态性丰富。该体系在29个香蕉基因组中获得较好的扩增结果,可望在香蕉植物起源和进化研究中应用。

关键词: 香蕉, SRAP, 扩增条件优化

Abstract: In order to establish and optimize the SRAP molecular marker system in Musa spp.,the concentrations of Mg2+, dNTPs, Taq DNA polymerase, primers which affect the SRAP-PCR reactions were optimized. The optimum system was as follows: Mg2+ 2.5 mmol·mL-1, dNTPs 250 μmol·L-1, Taq DNA polymerase 1.0 U, primer 0.5 μmol·L-1, template DNA 20 ng, 10×PCR buffer 2.5 μL.The total volume of reaction was 25 μL. Amplications were carriyed out on 29 banana cultivars genome using this optimum system. The results showed that the system was steady and reliable and would be helpful to study origin and evolution of Musa spp..

Key words: Musa spp., SRAP, Optimization

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