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中国农学通报

中国农学通报 ›› 2011, Vol. 27 ›› Issue (22): 162-166.

所属专题： 园艺

• 林学 园艺 园林 • 上一篇    下一篇

十字花科蔬菜黑腐病菌（Xanthomonas campestris pv. Campestris）AFLP分析体系的建立及优化

翟文慧 贾春枫 周莹 黄金宝 刘凡 严红   

  • 收稿日期:2011-02-25 修回日期:2011-03-28 出版日期:2011-09-05 发布日期:2011-09-05
  • 基金资助:

    国家自然科学基金课题项目;北京市科技计划项目

Construction and Optimization of an efficient System for Xanthomonas campestris pv. Campestris AFLP Analysis

  • Received:2011-02-25 Revised:2011-03-28 Online:2011-09-05 Published:2011-09-05

PDF (PC)

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摘要：

以英国华威大学收集的十字花科蔬菜黑腐病菌野油菜黄单胞菌野油菜致病变种（Xanthomonas campestris pv.campestris）的6个生理小种菌株为材料，优化了该类菌种的AFLP分析体系包括DNA的提取、Mse I/EcoR I酶切时间、扩增反应体系的组成及染色方法等步骤，建立了一套适于该菌种的AFLP分析体系。该体系中各最优因素为：酶切体系为50 μL，模板DNA用量为600 ng，酶切体系中Mse I和EcoR I各加入5 U，酶切温度为37℃，反应时间为4~6 h；预扩增产物稀释10倍进行选择性扩增；检测方法为银染法，银染液中硝酸银含量为8 g/L且染色时间是15 min时效果最佳，该体系的建立为该类菌种的分子水平遗传多样性研究提供了技术支撑。

关键词: 残留动态, 残留动态, 烯唑醇, 安全性评价

Abstract:

6 isolates of Xanthomonas campestris pv. Campestris collected by University of Warwick was used as the materials. The affect factors of AFLP were optimized, which were DNA extraction method, digestion time of Mse I/EcoR I, compositions of PCR reaction system and staining method, and an efficient system for X. campestris pv. Campestris AFLP analysis was established. The optimized factors were: the digestion system was 50 μL, the DNA template was 600 ng, the Mse I and EcoR I were both 5 U, the temperature was 37℃, reaction time was 4-6 h, pre-amplified products were diluted 10-fold for next selective amplification, the detection method was silver staining (AgNO3 content was 8 g/L and staining time was 15 min or so). The system was a powerful support for molecular level of genetic diversity in X. campestris pv. Campestris.

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