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| 马铃薯致病疫霉EST-SSR PCR反应体系的优化 | |
| 引用本文： | 王佳楠,吕文河,金光辉,白雅梅,李文霞,韩英鹏.马铃薯致病疫霉EST-SSR PCR反应体系的优化[J].东北农业大学学报,2009,40(6). |
| 作者姓名： | 王佳楠 吕文河 金光辉 白雅梅 李文霞 韩英鹏 |
| 作者单位： | 1. 东北农业大学农学院,哈尔滨,150030 2. 黑龙江八一农垦大学农学院,黑龙江,大庆,163319 3. 东北农业大学资源与环境学院,哈尔滨,150030 |
| 基金项目： | 黑龙江省自然科学基金项目 |
| 摘 要： | 由致病疫霉(Phytophthora infestans Montagne de Bary)引起的马铃薯晚疫病是危害马铃薯生产的最严重病害。试验以致病疫霉菌株HH06-23为模板,以Pi08N为引物,研究致病疫霉EST-SSRPCR反应体系中各主要成分和退火温度对扩增结果的影响。优化后的PCR反应体系为:25ng模板DNA,0.5mmol·L-1dNTPs,2μL10×Buffer(Mg2+Free),1.75mmol·L-1MgCl2,15pmol引物,1.2UTaqDNA聚合酶,加ddH2O至20μL;同时确定引物退火温度为63℃。PCR体系稳定性试验结果表明,优化后的致病疫霉EST-SSRPCR反应体系稳定、可靠,适合进行致病疫霉群体遗传多样性研究。 |
| 关 键 词： | 马铃薯 致病疫霉 EST-SSR PCR |
Optimization of PCR system in EST-SSR analysis of Phytophthora infestans |
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| WANG Jianan,LV Wenhe,JIN Guanghui,BAI Yamei,LI Wenxia,HAN Yingpeng.Optimization of PCR system in EST-SSR analysis of Phytophthora infestans[J].Journal of Northeast Agricultural University,2009,40(6). | |
| Authors: | WANG Jianan LV Wenhe JIN Guanghui BAI Yamei LI Wenxia HAN Yingpeng |
| Institution: | 1. College of Agriculture;Northeast Agricultural University;Harbin 150030;China;2. College of Agriculture;Heilongjiang August First Land Reclamation University;Daqing Heilongjiang 163319;3. College of Resource and Environment;China |
| Abstract: | Late blight caused by the oomycete Phytophthora infestans Montagnede Bary was a devastating disease on potato production. A total of 6 parameters affecting the PCR system in P. infestans were investigated based on the template DNA of the isolate HH06-23 and EST-SSR primer pair Pi08N. The results showed that the optimal annealing temperature was 63℃, and the optimum PCR system of EST-SSR were: 25 ng template DNA, 0.5 mmol·L-1 dNTPs, 2 μL 10×Buffer (Mg2+), 1.75 mmol·L-1 MgCl2, 15 pmol primer, 1.2 U Taq DNA poly-merase in total 20 μL reaction system. In addition, using the optimal PCR system, a total of 20 isolates of P. infestans were used for testing the stability and polymorphism of the PCR amplification. The clarity and abundant polymorphism indicated that this system was stable and suitable for study of the genetic diversity of P. infestans population. |
| Keywords: | EST-SSR PCR |
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