AMB Volume 40, Issue 4, 2024 Pages 446-456 https://doi.org/10.59393/amb24400404
Molecular Identification and Phylogenetic Analysis of Fusarium spp. Associated with Barley Grains Using DNA Barcoding in Bulgaria
Stoeva D., Gencheva D., Radoslavov G., Hristov P., Beev G.
This study examined the genetic relationships and distribution frequencies of Fusarium species isolated from barley in Bulgaria, using DNA barcoding and phylogenetic analysis. Molecular identification targeted ITS and β-tubulin regions, revealing seven Fusarium species. With 99.2–100% sequence similarity to reference Fusarium strains in GenBank, validating the accuracy of ITS and β-tubulin loci for distinguishing species. Among the identified species, F. sporotrichioides was the most frequent, with a prevalence of 46.1%. F. proliferatum followed, showing a lower but notable frequency of 15.4%, while F. poae and F. verticillioides each appeared in 8% of the isolates. Other species, including F. tricinctum and F. avenaceum, were less prevalent. Phylogenetic analyses, constructed using the Neighbor-Joining method, revealed geographic clustering trends. Bulgarian F. sporotrichioides isolates clustered closely with Chinese strains in the ITS phylogeny, suggesting a homogeneous population with ties to Asian origins. In contrast, F. avenaceum showed genetic diversity, clustering with both Chinese and Polish strains. F. brachygibbosum, a novel find in Bulgarian barley, aligned genetically with Russian strains, indicating potential cross-regional exchange. The β-tubulin-based phylogeny presented some distinctions from ITS, as F. sporotrichioides clustered with Russian and Finnish strains, underscoring the value of multi-locus barcoding for comprehensive phylogenetic insights. These findings highlight the diverse origins of Fusarium species in Bulgarian barley, which carry implications for disease management in the context of evolving climate conditions.
Keywords: Fusarium spp., barley, DNA barcoding, phylogeny, β-tubulin
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