ISSR analysis of Fusarium oxysporum Schl. in Hebei province

An Orthogonal design is applied to the optimization of ISSR-PCR reaction system for Fusarium oxysporum Schl. The effect of the five main factors was tested. A most suitable ISSR-PCR reaction system for Fusarium oxysporum Schl. was established. The optimized reaction system consists of 20 ng template DNA, 1 U Taq DNA polymerase, 0.4 μmol/L primer, 0.2 mmol/L dNTPs, 4.0 mmol/L Mg2+ and 2.5 μL 10× buffer. Optimal annealing temperature was decided by different primers. 2 primers with stable amplification and rich polymorphism for ISSR were selected. Based on the optimizing, 30 isolated strains from Hebei were tested in order to analyse the genetic diversity between of Fusarium oxysporum Schl. The isolated 30 strains were clustered into 2 genetic line-ages at 0. 86 genetic similarity. The genetic lineages of 30 strains showed no obvious relation with their geographical originals.