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中国农学通报

中国农学通报 ›› 2011, Vol. 27 ›› Issue (22): 219-223.

所属专题： 园艺

• 林学 园艺 园林 • 上一篇    下一篇

番石榴SRAP反应体系的建立与正交优化

杨祥燕 蔡元保 陈豪军 甘卫堂 周全光 李莲英 黄小江   

  • 收稿日期:2011-04-25 修回日期:2011-05-16 出版日期:2011-09-05 发布日期:2011-09-05
  • 基金资助:

    广西壮族自治区直属公益性科研院所基本科研业务费专项资金项目

Orthogonal Optimization and Establishment of SRAP-PCR Amplification System of Guava (Psidium guajava L.)

  • Received:2011-04-25 Revised:2011-05-16 Online:2011-09-05 Published:2011-09-05

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摘要：

采用正交设计方法，对影响番石榴SRAP反应体系的Mg2+、dNTPs、引物、Taq DNA聚合酶和模板DNA浓度等进行了优化，建立了适用于番石榴的SRAP反应体系。该优化的20 μL反应体系中包含2.5 mmol/L Mg2+，0.15 mmol/L dNTPs，0.4 μmol/L引物，1.5 U Taq DNA聚合酶和20 ng模板DNA。利用该优化体系通过64对SRAP引物组合对5份番石榴材料进行了SRAP-PCR扩增，结果表明SRAP引物及优化后的反应体系能够有效地用于番石榴种质资源鉴定及遗传多样性分析等研究。

关键词: 烤烟, 烤烟, 采收时间, 中性致香物质, 感官质量

Abstract:

Orthogonal design was applied to optimize SRAP-PCR system of Guava (Psidium guajava L.). Five main factors influencing on SRAP-PCR, including Mg2+, dNTPs, primer, Taq DNA polymerase and DNA template were examined by using orthogonal design. The results showed that the optimum concentration was Mg2+ 2.5 mmol/L, dNTPs 0.15 mmol/L, primer 0.4 μmol/L, Taq DNA polymerase 1.5 U, 20 ng template DNA with a total volume of 20 μL reaction solution, which suitable for P. guajava SRAP-PCR amplification. The molecular identification of 5 varieties of P. guajava was conducted by 64 SRAP primer combinations with the optimal SRAP marker system. It showed that the SRAP primers and its optimized system could be applied in the germplasm identification and genetic diversity analysis of P. guajava effectively.