RESUMEN
Species of the genus Alternaria are ubiquitous and frequently isolated from various plants, including crops. There are two phylogenetically and morphologically close Alternaria sections: the relatively well-known Infectoriae and the rarely mentioned Pseudoalternaria. Currently, the latter includes at least seven species that are less studied and sometimes misidentified. To perform precise identification, two primers (APsF and APsR) were designed and a sect. Pseudoalternaria-specific PCR method was developed. Thirty-five Russian A. infectoria-like strains were then examined. Five strains were found to be the members of the sect. Pseudoalternaria. Additionally, specificity of the previously developed primer set (Ain3F and Ain4R) was checked. It was found to be highly specific for sect. Infectoriae and did not amplify sect. Pseudoalternaria DNA. Identification of strains of the sect. Pseudoalternaria was supported and refined by phylogenetic reconstruction based on analysis of two loci, the glyceraldehyde-3-phosphate dehydrogenase gene (gpd), and the plasma membrane ATPase gene (ATP). These fungi belonged to Alternaria kordkuyana and A. rosae, which were the first detection of those taxa for the Eastern Europe. Alternaria kordkuyana was isolated from cereal seeds and eleuthero leaves. Alternaria rosae was obtained from oat seed. All strains of sect. Pseudoalternaria were not able to produce alternariol mycotoxin, as well as the majority of A. sect. Infectoriae strains.
RESUMEN
Production of type A trichothecenes has been reported in the closely related species Fusarium langsethiae and F. sporotrichioides. Here, we characterized a collection of Fusarium isolates from Siberia and the Russian Far East (hereafter Asian isolates) that produce high levels of the type A trichothecene T-2 toxin and are similar in morphology to the type A trichothecene-producing F. langsethiae, and to F. poae which often produces the type B trichothecene nivalenol. The Asian isolates possess unique macroscopic and microscopic characters and have a unique TG repeat in the nuclear ribosomal intergenic spacer (IGS rDNA) region. In Asian isolates, the TRI1-TRI16 locus, which determines type A versus type B trichothecene production in other species, is more similar in organization and sequence to the TRI1-TRI16 locus in F. sporotrichioides and F. langsethiae than to that in F. poae. Phylogenetic analysis of the TRI1 and TRI16 gene coding regions indicates that the genes in the Asian isolates are more closely related to those of F. sporotrichioides than F. langsethiae. Phylogenetic analysis of the beta-tubulin, translation elongation factor, RNA polymerase II and phosphate permease gene sequences resolved the Asian isolates into a well-supported sister lineage to F. sporotrichioides, with F. langsethiae forming a sister lineage to F. sporotrichioides and the Asian isolates. The Asian isolates are conspecific with Norwegian isolate IBT 9959 based on morphological and molecular analyses. In addition, the European F. langsethiae isolates from Finland and Russia were resolved into two distinct subgroups based on analyses of translation elongation factor and IGS rDNA sequences. Nucleotide polymorphisms within the IGS rDNA were used to design PCR primers that successfully differentiated the Asian isolates from F. sporotrichioides and F. langsethiae. Based on these data, we formally propose that the Asian isolates together with Norwegian isolate IBT 9959 comprise a novel phylogenetic species, F. sibiricum, while the two subgroups of F. langsethiae only represent intraspecific groups.