| 摘要: |
| 【目的】建立并优化适合杜仲的AFLP技术体系，为进一步构建其遗传图谱及开展分子标记辅助育种奠定基础。【方法】对杜仲基因组DNA提取方法、酶切与连接体系、预扩增和选择性扩增程序的影响因素进行分析，并对适合杜仲AFLP分析的引物组合进行筛选。【结果】模板DNA的提取采用改进的CTAB法，酶切模板DNA用量500 ng,酶切时间6 h，连接反应时间6 h，预扩增产物最适稀释倍数30倍。同时筛选出E-AAG+M-CAA，E-AAG+M-CAT，E-AAG+M-CTA，E-AAG+M-CTG，E-ACA+M-CAA，E-ACA+M-CTA，E-ACC+M-CTG共7对适合杜仲AFLP分析的引物组合。【结论】经重复验证，建立的AFLP反应体系适用于杜仲的AFLP分析。 |
| 关键词: 杜仲 AFLP 体系优化 引物筛选 |
| DOI： |
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| 基金项目:国家自然科学基金项目（30771751） |
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| Establishment and optimization of AFLP reaction system in Eucommia ulmoides Oliv. |
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| Abstract: |
| 【Objective】The study intended to establish an AFLP reaction system and provide technical support for the construction of genetic linkage map and molecular markers assisted breeding of Eucommia ulmoides Oliv.【Method】The main factors including extraction of genomic DNA method,reaction system of digestion and ligation,pre-amplification and selective amplification processes were analyzed and the primer paris were screened.【Result】Genomic DNA was extracted with improved CTAB method,500 ng template DNA in the digestion system,the reaction time 6 hours,the ligation time 6 hours,30 times of dilution for the products of pre-amplification for selective amplification.And 7 pairs of primers(E-AAG+M-CAA，E-AAG+M-CAT，E-AAG+M-CTA,E-AAG+M-CTG，E-ACA+M-CAA，E-ACA+M-CTA，E-ACC+M-CTG) were selected from 64 pairs of primer combination for the AFLP reaction system of E.ulmoides Oliv.【Conclusion】The results show that it is feasible to use the reaction system constructed by the study to conduct the AFLP analysis of E.ulmoides Oliv. |
| Key words: Eucommia ulmoides Oliv. AFLP establishment and optimization of the system screening of primer pairs |
