A Comprehensive Genetic Analysis of Mycotoxin-Producing Penicillium expansum Isolated from River Water Using Molecular Profiling, DNA Barcoding, and Secondary Structure Prediction

Abstract

This study marks the first report on the genetic characterization of Penicillium expansum strain capable of mycotoxin production isolated from river water. Situated in Ganagalawanipeta village, Srikakulam, Andhra Pradesh, India, where river water serves as a vital resource, our investigation probed the presence of pathogenic opportunistic fungi adept at mycotoxin synthesis. Over six months, 30 samples were collected to assess their occurrence. This article revolves around the use of morphological traits for Penicillium genus identification. Precise species determination involved PCR analysis using universal primers ITS1 and ITS4, followed by sequence analysis through NCBI-BLASTn and the ITS2 database. The analysis indicated a striking 99.49% genetic similarity to Penicillium expansum isolate MW559596 from CSIR-National Institute of Oceanography, Goa, an Indian isolate, with a resultant 600-base pair fragment. This sequence was officially cataloged as OR536221 in the NCBI GenBank database. Sequence and phylogenetic assessments were conducted to pinpoint the strain and geographical origin. Notably, the ribosomal nuclear ITS region displayed significant inter- and intra-specific divergence, manifested in DNA barcodes and secondary structures established via minimum free energy calculations. These findings provide crucial insights into the genetic diversity and potential mycotoxin production of P. expansum isolates, shedding light on the environmental repercussions and health risks associated with river water contamination from agricultural and aquaculture effluents. This pioneering research advances our understanding of mycotoxin-producing fungi in aquatic environments and underscores the imperative need for water quality monitoring in regions reliant on such water sources for their sustenance and livelihoods.