Abstract: Thirty polymorphic simple sequence repeat (SSR) markers have been developed and characterized from RAPD product in Aspergillus niger in order to protect its natural resources. Polymorphisms of these RAPD-SSR markers were evaluated in a natural population of 25 strains collected from Shandong in China. The number of alleles (Na) per locus varied from 2 to 9. Observed (H O ) and expected (H E ) heterozygosities ranged from 0.56 to 0.96, and from 0.52 to 0.88, respectively. All the RAPD-SSR loci conformed to Hardy–Weinberg equilibrium after Bonferroni correction. They have showed sufficient level of polymorphisms to estimate the genetic diversity, population structure and species conservation in the A. niger. Five additional fungus species, Penicillium chrysogenum, Penicillium citrinum, Aspergillus flavus, Aspergillus ochratoxin and Aspergillus sulphureus were assessed for cross-species amplification. Three of the five species showed at least three polymorphic loci. In addition, eight loci were found to be polymorphic in at least one species.
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