Abstract
<title>Abstract</title> <p> <bold>Background:</bold> Accurate fungal identification and cultivation are critical across clinical, agricultural, and industrial sectors. Misidentification of macrofungi can lead to fatal poisonings, while improper isolation of microfungi may result in misdiagnoses, crop losses, or contamination of commercial strains. <bold>Objectives:</bold> This systematic review synthesizes evidence on (1) diagnostic frameworks for fungal identification, (2) lethal fungal profiles and taxonomic differentiation, (3) formulation principles and efficacy of growth media, and (4) operational complications in mycological cultivation with corresponding mitigation strategies. <bold>Methods:</bold> A systematic literature search was conducted across PubMed, Web of Science, and Scopus databases for publications from 1986 to 2019. Studies were included if they addressed fungal identification protocols, growth media formulations, or cultivation complications. Data extraction focused on diagnostic accuracy, media performance metrics, and operational parameters. <bold>Results:</bold> A three-tiered diagnostic framework (DNA sequencing, macroscopic examination, and microscopic analysis) demonstrates the highest accuracy, with ITS region sequencing achieving species-level confirmation ¹. Fifteen lethal amatoxin-containing species were identified, with <italic>Amanita phalloides</italic> implicated in the majority of fatal ingestions ⁵,⁸,¹⁶. Standardized media (SDA, PDA, CMA, CZA, MEA) provide consistent benchmarks ¹⁰,¹¹, while sustainable alternatives including Rice Bran Dextrose Agar (RBDA) and Tomato Juice Agar (TJA) show comparable or superior biomass yields ². Thermal autoclaving artifacts, including Maillard reactions and acid hydrolysis, significantly impact media performance ³, while biological contamination and chemical selection bias remain critical operational challenges. <bold>Conclusions:</bold> Molecular methods are increasingly essential for reliable species confirmation, particularly given phenotypic variability among look-alike species ¹,¹⁹. Sustainable plant-based media offer viable low-cost alternatives ², though rigorous control of operational variables remains paramount for reproducible cultivation outcomes ³. </p>