Rapid and Efficient Method for DNA Extraction from Fungi of the Genus Morchella Dill. ex Pers.

Abstract

The taxonomic features of true morels (genus Morchella, Ascomycota) have long been a difficult problem because the identification of species of such a taxonomically complex genus is impossible without the use of molecular phylogenetics methods. To date, there are a large number of DNA extraction methods, but the search for a fast and efficient method of DNA extraction from various organisms, including fungi, remains an urgent problem in modern biological research. Methods. The study material consisted of the fruiting bodies of fungi belonging to the genus Morchella. Total DNA extraction was performed using the original DNA Microprep Isolation from Plants protocol (DNA Microprep method). For comparison, the modified CTAB method was also used to extract DNA from all samples. Extraction efficiency was determined by spectrophotometry. The ITS marker sequence of the ribosomal RNA nuclear gene cluster was amplified using primers ITS1 and ITS4, and the amplification products were separated by electrophoresis in an agarose gel. Results. DNA-containing pellets were formed in all samples extracted by the DNA Microprep method, as confirmed by spectrophotometric analysis. The extracted nucleic acid was of acceptable quantity and purity. The amplification of the ITS marker sequence was successful for all samples isolated by the DNA Microprep method, resulting in amplicons of different lengths, which may indicate a potential difference in the taxonomic affiliation of the studied fruiting bodies of morels. Regarding PCR products based on DNA extracted by the CTAB method, 80% of the samples tested were positive for PCR amplification. Conclusions. The study suggests that the DNA Microprep method is a convenient and effective way to extract DNA from morels' fruit bodies. The method offers advantages such as a short extraction process, availability of reagents, and environmental friendliness. Additionally, PCR products were successfully obtained from the extracted DNA. To accurately identify the species of the taxonomically complex genus Morchella, it is necessary to determine the nucleotide sequences of the obtained amplicons and perform a phylogenetic analysis.