中文摘要：

为建立并优化适合向日葵的Pst Ⅰ和Mse Ⅰ组合的AFLP技术体系,为下一步构建遗传连锁图谱及开展分子标记辅助育种奠定基础,对向日葵基因组DNA提取方法、酶切连接体系和选择性扩增程序的影响因素进行优化,并对适合向日葵AFLP分析的引物组合进行筛选。结果表明,模板DNA的提取采用改进CTAB法,酶切连接采用一步法反应14 h,在选择性扩增体系20μL中,Mg2＋和d NTP浓度分别为2 mmol/L和0.3 mmol/L,选扩引物的浓度为0.6 mmol/L,预扩增产物最适稀释倍数为30倍。同时筛选出了稳定且多态性丰富的48对适合向日葵AFLP分析的引物组合。

英文摘要：

To provide technical support for construction of genetic linkage map and molecular markers assisted breeding of sunflower, an optimized AFLP reaction system using double-digestion of Pst I and Mse I endonncleasc has been established. The extraction method of genomic DNA, the DNA digestion and ligation system and several key factors affecting the PCR selective amplification are optimized. In addition, the primer combination which suits for analysis of AFLP is also screened in sunflower. The results show that the high quality of genomic DNA as PCR template can he extracted with the improved CTAB method, and that the DNA digestion and ligation can be used as a one-step process with the reaction time of 14 hours. In the optimized selection amplification system of total volume of 20 IxL, the Mg2＋ concentration is 2 mmol/L, dNTPs concentration is 0.3 mmol/L, the primers concentration is 0.6 mmol/L and the products of pre-amplification for selective amplification are diluted to 30 times. Moreover, 48 pairs stable and polymorphisms primer have been selected for AFLP analysis in sunflower.