李种质资源ISSR反应体系的建立

Establishment of the Inter-simple Sequence Repeats Reaction System in Plum Germplasm

以5′-(AC)_9 A-3′为引物对牛心李(Prunus salicina cv.Niuxinli)ISSR(inter-simple sequence repeats)反应体系的优化研究表明,25μL ISSR反应体系中,Taq酶、Mg^(2+)、引物、模板DNA和dNTPs5种主要成分的最适浓度分别是:0.3U、1.5mmol/L、0.2μmol/L、20ng和0.16mmol/L。利用该优化体系,以5′-(AC)_9 T-3′为引物,构建了中国李18个品种的ISSR指纹图谱,该引物可将这些品种完全区别开来;以5′-(AC)_9 C-3′为引物,构建了李属6类种质资源的ISSR指纹图谱,该引物区分率为100％。

The optimal reaction of ISSR (Inter-simple Sequence Repeats) in Prunus salicina cv. Niuxinli was studied with primer 5'-(AC)9A-3'. The results showed that the optimum concentration of five important components i.e. Taq DNA poly-merase, Mg2+, primer, template DNA and dNTPs in 25 μX reaction system of ISSR were 0.3U, 1.5 mmol/L, 0.2 μmol/L, 20 ng and 0.16 mmol/L, respectively. The ISSR fingerprinting of 18 Japanese plum cultivars and 12 plum species or variety were established by using optimum system with primer 5'-(AC)9T-3'and 5'-(AC)9C-3', respectively. The identification rate of plum cultivars and species or variety was 100%.