RESUMO
Ice from Arctic glaciers contains large populations of yeasts. We studied 38 isolates from this environment, which were initially identified as Debaryomyces sp. related to Debaryomyces hansenii by sequence analysis of the D1/D2 domains of 26S rDNA. An analysis of the distribution of mitochondrial DNA insertions in the nuclear genome showed that 25 of these isolates were related to, but distinct from, D. hansenii. Sequence analysis of the ACT1 gene of these 25 isolates revealed that they formed three different types of putative hybrids. In particular, 23 putative hybrids carried an ACT1 sequence identical to that of three Debaryomyces strains, CBS 790, CLIB 660, CLIB 949, previously classified as associated with D. hansenii and an ACT1 sequence of an undescribed taxon. The latter sequence displayed between 22 and 27 bp divergence (2.6-3.2 %) over 841 bp from sequences of closely related Debaryomyces sp., suggesting that this new taxon very likely represents a novel species for which no pure strain is available. Sequence comparisons of CBS 790, CLIB 660, and CLIB 949 with related Debaryomyces type strains also revealed an important sequence divergence. The putative hybrids described in this study could be differentiated from non-hybrid isolates and other Debaryomyces species on the basis of their use of a number of carbon sources.
Assuntos
Variação Genética , Camada de Gelo/microbiologia , Saccharomycetales/classificação , Saccharomycetales/genética , Actinas/genética , Regiões Árticas , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Dados de Sequência Molecular , Filogenia , RNA Ribossômico/genética , Recombinação Genética , Saccharomycetales/isolamento & purificação , Análise de Sequência de DNARESUMO
Seven apiculate yeast strains that were isolated from the flowers of Syphocampylus corymbiferus Pohl in Brazil are genetically, morphologically and phenotypically distinct from recognized species of the genera Hanseniaspora and Kloeckera. Genetic discontinuities between the novel strains and their closest relatives were found using a networking approach based on the concatenated sequences of the rRNA gene (internal transcribed spacer and D1/D2 of the LSU), and the protein-coding genes for actin and translation elongation factor-1α. Phylogenetic analysis based on the rRNA and the actin gene placed the novel species represented by the strains in close relationship to Hanseniaspora meyeri and Hanseniaspora clermontiae. PCR fingerprinting with microsatellite primers confirmed the genetic heterogeneity of the novel species. The name Hanseniaspora nectarophila sp. nov. is proposed, with UFMG POG a.1(T) (â= ZIM 2311(T)â = CBS 13383(T)) as the type strain; MycoBank no. MB807210. As the current description of the genus does not allow the presence of multilateral budding, an emended diagnosis of the genus Hanseniaspora Zikes is proposed.
Assuntos
Campanulaceae/microbiologia , Flores/microbiologia , Hanseniaspora/classificação , Filogenia , Actinas/genética , Brasil , DNA Fúngico/genética , DNA Espaçador Ribossômico/genética , Genes Fúngicos , Hanseniaspora/genética , Hanseniaspora/isolamento & purificação , Dados de Sequência Molecular , Técnicas de Tipagem Micológica , Fator 1 de Elongação de Peptídeos/genética , Análise de Sequência de DNARESUMO
A polyphasic molecular approach was used in order to characterize and taxonomically assign Debaryomyces yeast isolates of different origins. Actin 1 (ACT1) gene sequences coupled with AFLP markers showed that the investigated yeasts belonged to the recently reinstated species D. hansenii, D. fabryi and D. tyrocola. The strain HA1179 was supposed to be a D. hansenii strain with introgressed D. fabryi DNA segments. This strain acquired ribosomal RNA encoding genes (rDNA) and the ACT1 gene from the species D. fabryi and D. hansenii respectively. Comparative sequence analysis of the ACT1 gene, ITS1-5.8S-ITS2 (5.8S-ITSs) and D1/D2 regions, suggested that five strains isolated from a municipal wastewater treatment plant could represent a new taxon of the genus, for which the name Debaryomyces vindobonensis was proposed. The calculated degree of similarity between the AFLP patterns indicated that the strains of D. vindobonensis and the closely related species were separated by the values ï¼0.5. New yeast isolates showed very similar morphological and physiological properties to related Debaryomyces species. They differed notably only by the assimilation of rhamnose and growth at 50% glucose. In contrast to the other species, D. vindobonensis was unable to assimilate starch.
Assuntos
Saccharomycetales/classificação , Saccharomycetales/isolamento & purificação , Águas Residuárias/microbiologia , Actinas/genética , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Dados de Sequência Molecular , Filogenia , Saccharomycetales/genética , Análise de Sequência de DNARESUMO
Two protein-coding genes, actin and translation elongation factor-1alpha (EF-1alpha), as well as two ribosomal gene regions, D1/D2 domains of the large subunit and both internal transcribed spacers including the 5.8S gene region, were evaluated regarding their usefulness for reconstruction of phylogenetic relationships in the Hanseniaspora-Kloeckera species group. This included analyses of sequence divergence values, heterogeneity of evolutionary rates and the reliability of the inferred trees. Both protein-coding genes showed greater capacities to resolve at the strain level and between the closely related species of Hanseniaspora-Kloeckera, compared with the ribosomal gene regions. However, to obtain a fully resolved and reliable phylogenetic tree that reflected the biological relationships it was necessary to combine three congruent sequence datasets. The novel species Hanseniaspora pseudoguilliermondii sp. nov. (type strain CBS 8772T) is described as a result of the application of various molecular approaches to delimit species. Furthermore, incongruent gene genealogies of genetically divergent strains of Hanseniaspora occidentalis, as determined by amplified fragment length polymorphism analysis and DNA-DNA reassociation measurements, indicated the presence of two novel varieties, H. occidentalis var. occidentalis (type strain CBS 2592T) and H. occidentalis var. citrica var. nov. (type strain CBS 6783T), which could be distinguished by habitat preference.
Assuntos
Saccharomycetales/classificação , Actinas/genética , Impressões Digitais de DNA , DNA Fúngico/química , DNA Fúngico/isolamento & purificação , DNA Ribossômico/química , DNA Ribossômico/isolamento & purificação , DNA Espaçador Ribossômico/genética , Proteínas Fúngicas/genética , Genes de RNAr , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Fator 1 de Elongação de Peptídeos/genética , Filogenia , Polimorfismo de Fragmento de Restrição , RNA Fúngico/genética , RNA Ribossômico/genética , RNA Ribossômico 5,8S/genética , Saccharomycetales/genética , Análise de Sequência de DNARESUMO
Recent epidemiological evidence suggests that there are two forms of Crohn disease (CD): perforating and nonperforating. We hypothesized that, just as with tuberculoid and lepromatous leprosy, differences in the two forms of CD would be both identified and determined by differences in the host immune response. Resected intestinal tissue from control patients as well as perforating and nonperforating CD patients was evaluated for mRNA levels. We employed 32P PCR amplification with published or custom-designed primers of a housekeeping gene (beta-actin); a human T-cell marker (CD3-delta); and the cytokines tumor necrosis factor alpha, transforming growth factor beta, granulocyte/macrophage colony-stimulating factor, interleukin (IL) 1 beta, IL-1ra, and IL-6. Differences were identified with IL-1 beta (control = 162 +/- 57 vs. perforating = 464 +/- 154 vs. nonperforating = 12,582 +/- 4733; P < or = 0.02) and IL-1ra (control = 1337 +/- 622 vs. perforating = 2194 +/- 775 vs. nonperforating = 9715 +/- 2988; P < or = 0.02). These data corroborate the epidemiological observation that there are two forms of CD. Nonperforating CD, the more benign form, is associated with increased IL-1 beta and IL-1ra mRNA expression. We conclude that it is the host immune response that determines which form of CD becomes manifest in any given individual and discuss the investigative, diagnostic, and therapeutic implications of these observations.