Abstract
Rose is one of the most important cultivated ornamental plants in the world. A molecular approach using inter-simple sequence repeat (ISSR) markers was applied to seven species of Rosa . To obtain clear and reproducible bands on 2% agarose gels, 9 ISSR primers and 5 parameters (annealing temperature, DNA concentrations, primer concentrations, Taq DNA polymerase and MgCl 2 concentrations) were screened. The resolution of six ISSR markers was performed, with optimal annealing temperature (Ta) varying from 45 to 50°C. A total of 66 DNA fragments were amplified, of which 50 were polymorphic. The optimal conditions for ISSR system were determined as follows: MgCl 2 concentration was 2 mM, the quantity of Taq DNA polymerase 1 U, template DNA 30 ng and the concentration of primer was 1 μM and the reaction program was: initial denaturation for 5 min at 94°C, 35 cycles of denaturation for 30 s at 94°C, annealing for 45 s at specific annealing temperature for each primer, extension for 2 min at 72°C and a final 10 min extension at 72°C. Key words : Inter-simple sequence repeat marker, rose species, genetic diversity, optimization.
Published Version
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