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| 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 | |
| 引用本文： | 翟文慧,贾春枫,周莹,黄金宝,刘凡,严红.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[J].中国农学通报,2011,27(22):162-166. |
| 作者姓名： | 翟文慧 贾春枫 周莹 黄金宝 刘凡 严红 |
| 作者单位： | 1. 北京市农林科学院植物保护环境保护研究所,北京,100097 2. 青岛出入境检验检疫局,山东青岛,266001 3. 北京市农林科学院蔬菜研究中心,北京,100097 |
| 基金项目： | 国家自然科学基金课题项目“花椰菜-黑芥抗病异附加系的培育与鉴定”(30771206); 北京市科技计划项目“农村科技协调员支撑服务工程”(Z09090501040904-4) |
| 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 |
| 收稿时间： | 2011/2/25 0:00:00 |
| 修稿时间： | 2011/3/28 0:00:00 |
Construction and Optimization of an efficient System for Xanthomonas campestris pv. Campestris AFLP Analysis |
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| Zhai Wenhui,Jia Chunfeng,Zhou Ying,Huang Jinbao,Liu Fan,Yan Hong.Construction and Optimization of an efficient System for Xanthomonas campestris pv. Campestris AFLP Analysis[J].Chinese Agricultural Science Bulletin,2011,27(22):162-166. | |
| Authors: | Zhai Wenhui Jia Chunfeng Zhou Ying Huang Jinbao Liu Fan Yan Hong |
| Institution: | Zhai Wenhui 1,Jia Chunfeng 2,Zhou Ying 1,Huang Jinbao 1,Liu Fan 3,Yan Hong 1(1 Institute of Plant and Environment Protection,Beijing Academy of Agriculture and Forestry Sciences,Beijing 100097,2 Qingdao Entry-exit Inspection and Quarantine Bureau,Qingdao Shandong 266001,3 Vegetable Research Center,Beijing 100097) |
| 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（AgNO 3 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. |
| Keywords: | crucifers black rot Xanthomonas campestris AFLP |
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