Enhanced DNA Extraction From Dimorphic Fungi: Exploring the Advantages of Blastospores Versus Mycelia.

Abstract

The extraction of high-quality DNA from fungi remains a significant challenge due to the structural complexity and resilience of fungal cell walls. This study presents the first comprehensive comparison of DNA extraction efficiency between blastospores (BS) and mycelial cells (MCs) in dimorphic fungi. We evaluated six DNA extraction procedures and two pre-treatment methods to determine their effectiveness in extracting DNA of high quality and yield from both forms. DNA quantification was performed using absolute RT-qPCR targeting the single-copy γ-Actin gene, with specificity and efficiency validated through standardized protocols. Our results showed that BS consistently yielded higher DNA quantity and quality across all conditions, with a mean antilog DNA copy concentration of 5.03 ± 0.20, compared to 2.51 ± 0.20 for MC. DNA quality was superior in BS, as indicated by significantly better 260/280 and 260/230 ratios (31.5% vs. 7.4% for MC, p value = 0.002). Additionally, BS produced higher molecular weight DNA (10,770.5 base pairs vs. 8139.0 base pairs for MC, p value < 0.001) and exhibited greater long-term stability at both 4°C and -20°C. The results demonstrate that BS serve as a more reliable source of high-quality DNA compared to MC. This finding enhances current methodologies in molecular biology and supports more accurate analyses, particularly in studies involving dimorphic fungi.