DNA Analysis of Five Microalgae Species Using Molecular Markers

Background: Microalgae are important for biotechnology and environmental monitoring, but species identification is difficult because many taxa share similar morphology and life cycles. This study aimed to assess genetic diversity and phylogenetic relationships of five microalgae species using ISSR markers.Methods: Anabaena sp., Nostoc sp., Microcystis sp., Chlorella sp., and Scenedesmus sp. were cultivated under laboratory conditions. Genomic DNA was isolated using CTAB-based extraction, quantified by Microcystis NanoDrop spectrophotometry, and evaluated by agarose gel electrophoresis. ISSR-PCR was performed using the dinucleotide repeat primers (AG)9C and (GA)9C. Amplified fragments were separated on agarose gels, compared using VisionCapt software, and used to calculate Nei’s genetic distance and construct a phylogenetic tree with POPGENE v.1.32.Result: Gel electrophoresis confirmed that the isolated DNA was of sufficient quality for PCR analysis. The (AG)9C primer generated polymorphic profiles, with Anabaena sp. and Nostoc sp. sharing bands at 750, 950, 1000, and 1150 bp, indicating close genetic similarity. Microcystis sp., Chlorella sp., and Scenedesmus sp. showed distinct fragment patterns. The (GA)9C primer also revealed interspecific variation, producing species-specific bands, while no visible amplified fragment was detected in Scenedesmus sp. Phylogenetic analysis grouped Anabaena sp. and Nostoc sp. together, while the remaining species formed separate branches.Conclusion: ISSR-PCR using (AG)9C and (GA)9C primers effectively detected genetic diversity among the studied microalgae. The generated DNA fragments are useful molecular markers for biodiversity assessment, taxonomic identification, and future DNA barcoding studies. Integrating ISSR-PCR with sequencing approaches may strengthen microalgae classification and support biotechnological applications.KeywordsMicroalgae, RAPD analysis, ISSR analysis, Genetic diversity, Metagenomics, Phylogenetic analysis