青花菜SRAP-PCR体系优化与品种分子鉴定

Optimization of SRAP-PCR System and Cultivar Molecular Identification in Broccoli(Brassica oleracea var. italica)

本试验对青花菜基因组DNA的SRAP-PCR体系中主要影响因子Mg2+、dNTPs和引物浓度进行了优化。结果表明,反应体系中适宜浓度为Mg2+1.5-3.0mmol/L,dNTPs0.4mmol/L,引物0.25-0.50μmol/L(模板DNA约20ng,16μL反应体系)。运用优化的反应体系,对20个青花菜品种进行分子鉴定,从10个引物组合中筛选到7个多态性引物组合,获得60个多态性位点,平均每个引物组合在供试品种中产生8.6个多态性位点,鉴别品种数4.3个。双引物组合me1/em6与me2/em9可以区分所有供试材料。

The concentration of Mg2＋,dNTPs and primer in the SRAP-PCR system was optimized for genomic DNA in broccoli.The optimized concentrations were Mg2＋ at 1.5-3.0 mmol/L,dNTPs at 0.4 mmol/L,primer at 0.25-0.50 μmol/L,with 20 ng DNA and 16 μL PCR volume.The molecular identification of 20 broccoli cultivars were conducted with the SRAP optimized system,seven out of ten primer combinations detected 60 polymorphic loci,an average of 8.6 polymorphic loci and 4.3 distinguished cultivars by each primer combination.All the cultivars could be distinguished with the combination of me1/em6 and me2/em9 in the study.