Molecular Identification of Six Mushroom Species Using Random Amplified Polymorphic Dna (RAPD) and Internal-Transcribed Spacer (ITS) Primers

Molecular identification of six mushroom species was conducted using Random Amplified Polymorphic DNA (RAPD) primers and Internal Transcribed Spacer (ITS) primers. The six mushrooms were randomly collected from Lagos and Ogun State in Nigeria. The genomic DNA of the mushroom was extracted using SDS protocol and quantified spectrophotometically. The gel electrophoresis of genomic DNA was run on 1.5% Agarose gel and visualized using UV-Trans-illuminator. The rDNA-ITS fragment of the genomic DNA was amplified using two ITS primer combinations (ITS4A+ ITS and ITS4B + ITS). The Primers ITS4B and ITS amplified single monomorphic bands for all five Pleurotus spp except Ganoderma spp whereas ITS4A and ITSS only amplified two Pleurotus sp (Pleurotus saju-caju, Pleurotus ostreatus FIIRO) and Ganoderma spp. RAPD-PCR technique was used to reveal DNA polymorphism in DNA of the six fungi (five Pleurotus species and one Ganoderma spp) in order to search for the sources of differences that could be used as a DNA marker. In RAPD-PCR, six different decamer primers (OPK 01, OPD 04, OPD 7, OPD 11, OPM 14 and OPL 20) chosen randomly were employed to detect the genetic polymorphism among studied strains. A total of 206 amplicons were produced by these primers and out of which gave 164 bands for polymorphic bands with 42 bands recorded as monomorphic bands with an average percentage polymorphism of 83.5%. The polymorphism is high in OPK 01, OPD11 and OPL20 (100%) which make them transferable for diversity studies for genetic improvement. The identification characterization of the mushroom samples was analyzed of which genetic dissimilarity showed a range of 0.611 to 0.833. The highest was observed with RAPD primers OPK01, OPD11 and OPL20 (0.833) while OPD04 was lowest (0.611). The number of amplified fragments per primer ranged from 16 (OPD7) to 36 (OPK 01) with a polymorphic information content (PIC) ranging from 0.932 to 0.956.