Molecular Identification of Edible Mushroom Species Traded in Markets Across Anambra and Enugu States Using ITS Sequencing

Edible mushrooms are economically and nutritionally significant in Southeast Nigerian cuisine, yet species identification in market settings relies almost entirely on morphological assessment, which is unreliable for distinguishing edible from toxic species. Molecular barcoding provides a precise and reproducible alternative for mushroom authentication in food markets. This study applied internal transcribed spacer (ITS) sequencing for the molecular identification of wild mushrooms sold in markets across Awka and Enugu city, covering Anambra and Enugu States, Southeast Nigeria. Forty mushroom specimens were purchased from five markets per state. DNA was extracted from dried tissue using a commercial CTAB-based kit. PCR amplification of the ITS1-5.8S-ITS2 region was performed using universal fungal primers (ITS1F/ITS4), and amplicons were Sanger sequenced. Sequences were queried against the UNITE fungal reference database. Thirty-two of 40 specimens were successfully identified to species level. Identified species included Pleurotus ostreatus (35%), Lentinula edodes (22%), and Termitomyces robustus (18%). Importantly, two specimens could not be matched and showed sequence similarity to potentially toxic Amanita species, highlighting food safety risks. This study demonstrates the feasibility and importance of molecular authentication for edible fungi in Nigerian market chains. Keywords: edible mushrooms, ITS sequencing, DNA barcoding, food safety, Southeast Nigeria.