| 摘要: |
| [目的]建立悬铃叶苎麻植物AFLP反应体系,为其无融合生殖的分子标记及构建遗传图谱提供技术支持.[方法]以悬铃叶苎麻叶片为材料,采用改良CTAB法提取基因组DNA,并对酶切与连接体系、预扩和选扩程序进行探索,建立一套优化的AFLP反应体系.[结果]通过对EcoRⅠ+NNN/MseⅠ+NNN的64对引物组合的筛选,获得了清晰的选择性扩增图谱和多态性较高的引物组合.[结论]经多次重复验证,建立的优化体系可用于悬铃叶苎麻的AFLP分析. |
| 关键词: 悬铃叶苎麻 AFLP 扩增图谱 银染 无融合生殖 |
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| 基金项目:国家重大基础研究前期研究专项（2003CCA00400） |
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| Establishment of AFLP reaction system in Boehmeria tricuspis (Hance) Makino |
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| Abstract: |
| 【Objective】 The study presented intended to construct the AFLP reaction system of Boehmeria tricuspis(Hance) Makino,and provided technical support for molecular markers and genetic maps of apomixes.【Method】 Genomic DNA was extracted with improved CTAB method from Boehmeria tricuspis;some key factors affecting reaction system of digestion and connection,pre-amplification and selective amplification processes were studied and an optimized AFLP reaction system was established.【Result】 With this reaction system,the 64 primers of EcoR Ⅰ+NNN/Mse Ⅰ+NNN combinations were screened,clear DNA electrophoresis fingerprints and high polymorphism of primers' combinations of Boehmeria tricuspis were obtained and the reproducibility was high.【Conclusion】 Repeated results show that it is feasible to use the reaction system constructed by the study to conduct the AFLP analysis of Boehmeria tricuspis. |
| Key words: Boehmeria tricuspis (Hance) Makino AFLP amplification pattern silver staining apomixis |
