Search

Soil microorganisms play an important role in soil quality and they interact closely with vegetation. Little is known about yeast diversity and function in forest soil ecosystems and their interactions with other biotic soil components, particularly in the mycorrhizosphere. We studied the diversity of yeasts inhabiting the bulk-soil, rhizosphere and ectomycorrhizosphere of a Nothofagus pumilio forest in Nahuel Huapi National Park (Bariloche, Argentina). Ectomycorrhizal infection was observed in all N. pumilio trees studied. A total of 126 yeast isolates were obtained, including 18 known and three possibly new species. Basidiomycetous yeasts were predominant in all soil fractions, and the most frequently isolated species was Cryptococcus podzolicus. Diversity indices and multivariate analyses were used to study and compare yeast communities in the bulk-soil, rhizosphere and ectomycorrhizosphere. Yeasts able to ferment glucose were found associated with the rhizosphere. Many of the recovered yeast species were associated with lignocelluloses compound degradation, which suggest that yeast plays an important role as a decomposer in these forest soils. Each soil fraction has a distinct yeast assemblage related to their physiologic capacities and soil nutrient availability.

The synthesis of the UV Absorbing compounds named mycosporines (MYCs) has been detected in a few basidiomycetous yeast groups. Conspicuous accumulation of mycosporine-glutaminol-glucoside (MGG) in yeasts requires photo-induction and its photoprotective function has been postulated. The distribution of the ability to produce MYCs appeared to be related to the yeast taxonomic affiliation. In view of the potential significance of MYCs in yeast taxonomy, we here studied the distribution of this trait among dimorphic basidiomycetes of the Pucciniomycotina. Of the 94 fungal species (377 strains and 33 genera) tested, almost half were MYC-positive and MGG was the main compound produced. MGG synthesis was observed for representatives of five of seven Pucciniomycotina classes, indicating that this trait is widely distributed in this group. MGG detection proved useful for the differentiation of species of the polyphyletic genera, such as Rhodotorula and Sporobolomyces ,t hat are phylogenetically separated. MGG quantification and UV tolerance studies in Cystobasidiomycetes supported the idea that the habitat of origin of each strains is important in the level of MGG synthesis and that MYCs have a photoprotective function in yeasts. The taxonomic value of this trait in fungal systematics is discussed. Copyright  2011 John Wiley & Sons, Ltd.

Nahuel Huapi (NH) Lake is an oligotrophic temperate lake of glacial origin with high transparency, surrounded by well-developed forests and located at San Carlos de Bariloche, Nahuel Huapi National Park, in Patagonia, Argentina. In this lake, we characterized yeast distribution and diversity along a south-to-north transect and established a relationship between the ability to produce photoprotective compounds (PPCs) (carotenoid pigments and mycosporines) and the occurrence of yeast at different collection points. Subsurface water samples were filtered for yeast isolation. Total yeast counts ranged between 22 and 141 CFU L(-1) , and the highest values corresponded to the most impacted sites. Littoral sites had a low proportion of yeast-producing PPCs and this group prevailed in pelagic sites. This is probably a result of the high transparency of the water and the increased UV exposure. The yeast community from NH Lake showed a high species richness and a uniform distribution of taxa between pelagic and border collection points. Yeasts were identified as belonging to 14 genera and 34 species. Rhodotorula mucilaginosa and Cryptococcus victoriae were the most frequently found species, representing 14.4% and 13.6% of the total yeast isolates, respectively. Most of the yeast isolates demonstrated at least one extracellular enzymatic activity (mainly cellulase and lipase activities), which suggested that these microorganisms are metabolically active in the lake.

The occurrence and distribution of Xanthophyllomyces dendrorhous associated with Cyttaria hariotii parasitizing three Nothofagus species (N. dombeyi, N. antarctica and N. pumilio) in northwestern Patagonia (Argentina), as well as the factors that may affect this distribution were herein studied. Between 2000 and 2007, samples were obtained from 18 different locations. Based on physiological tests and morphological characteristics of sexual structures, 72 isolates were identified as X. dendrorhous. Representative strains were studied by MSP-PCR fingerprinting and sequence analysis of the ITS region. MSP-PCR fingerprints were similar for the newly isolated strains, and were also identical to the profiles of the strains previously found in this region. Patagonian strains appear to be a genetically uniform and distinct population, supporting the hypothesis that the association with different host species has determined genetically distinct X. dendrorhous populations worldwide. X. dendrorhous was recovered from N. dombeyi and N. antarctica. Approximately half the sampling sites and samples were positive for X. dendrorhous, but the isolation recovery rate was low. X. dendrorhous was absent in the early stages of ascostromata maturation, becoming more abundant in later stages. The present work represents a step forward in the understanding of the natural distribution and ecology of this biotechnologically relevant yeast. Se estudió la ocurrencia y la distribución de Xanthophyllomyces dendrorhous asociado a Cyttaria hariotii en tres especies de Nothofagus (N. dombeyi, N. antarctica y N. pumilio) del noroeste de la Patagonia (Argentina), y los factores que podrían afectar esta distribución. El muestreo se realizó entre 2000 y 2007 en 18 sitios diferentes. Según las pruebas fisiológicas y las características morfológicas de las estructuras sexuales, 72 de los aislamientos obtenidos se identificaron como X. dendrorhous. Se estudiaron cepas representativas mediante la técnica de MSP-PCR fingerprinting y secuenciación de la región ITS. Los perfiles de MSP-PCR fueron similares, tanto entre los nuevos aislamientos como entre estos y los de cepas previamente obtenidas en la región. Aparentemente, las cepas patagónicas forman una población genéticamente uniforme y distinta de otras poblaciones. Esto apoya la hipótesis de que la asociación con diferentes especies hospedadoras ha determinado la diferenciación genética de X. dendrorhous en todo el mundo. X. dendrorhous se recuperó de N. dombeyi y de N. antarctica. Aproximadamente la mitad de los sitios de muestreo y de muestras fueron positivos para X. dendrorhous, pero la tasa de aislamiento fue muy baja. X. dendrorhous está ausente en estadios tempranos de maduración de ascostromas y se hace más abundante en estadios más tardíos. El presente trabajo contribuye al mejor entendimiento de la distribución natural y la ecología de esta levadura, de relevancia biotecnológica. Fil: Libkind Frati, Diego. Universidad Nacional del Comahue. Centro Reg.universidad Bariloche. Departamento de Biologia. Laboratorio de Microbiologia Aplicada y Biotecnologia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Patagonia Norte. Instituto de Investigación en Biodiversidad y Medioambiente; Argentina Fil: Tognetti, Celia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional del Comahue. Centro Reg.universidad Bariloche. Departamento de Biologia. Laboratorio de Microbiologia Aplicada y Biotecnologia; Argentina Fil: Ruffini, Alejandra. Universidad Nacional del Comahue; Argentina. Universidad Nacional del Comahue. Centro Reg.universidad Bariloche. Departamento de Biologia. Laboratorio de Microbiologia Aplicada y Biotecnologia; Argentina Fil: Sampaio, José Paulo. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Centro de Recursos Microbiológicos ; Portugal Fil: Giraudo, Maria Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional del Comahue. Centro Reg.universidad Bariloche. Departamento de Biologia. Laboratorio de Microbiologia Aplicada y Biotecnologia; Argentina

The ability of the basidiomycetous yeast Xanthophyllomyces dendrorhous (Phaffia rhodozyma) to accumulate astaxanthin is responsible for the industrial use of this yeast as a microbial source of pigments for aquaculture. It is also hypothesized that astaxanthin accounts for its ability to thrive in highly oxidative and UV-exposed habitats. Here, we assessed the ability of this species to synthesize UV-absorbing compounds generally known as mycosporines, evaluated the effect of culture media in the production of these compounds and compared its UV growth resistance and tolerance with other yeasts. The 48 wild and collection strains screened were positive for mycosporines and a unique compound identified as mycosporine-glutaminol-glucoside (MGG) was detected. Thus, the ability of X. dendrorhous to produce MGG, as described here for the first time, is so far unique among the Cystofilobasidiales. The compound was synthesized constitutively, although growth under visible light and, to a greater extent, UVA radiation stimulated its production. Strains from UV-exposed habitats produced larger quantities and oligotrophic complex media seemed to favor MGG accumulation. UV tolerance and survival of X. dendrorhous was high and comparable to that of the polyextremophilic Rhodotorula mucilaginosa. The taxonomical and ecological implications of the production of MGG by X. dendrorhous are discussed.

In this paper we report the relationship between carotenoids and ergosterol and cell UV-B resistance in different strains of the yeast Rhodotorula mucilaginosa. Cell survival was studied using a set of 13 strains; additionally, two mutants (a hyper-producing one and a colourless one) in combination with diphenylamine (DPA), a carotenogenesis inhibitor, were used. A positive correlation between total carotenoids and survival to UV-B radiation was found. However, when individual carotenoid concentrations were tested, only torularhodin was found to be significantly related to UV-B survival. On the contrary, ergosterol did not affect survival. The hyper-pigmented strain showed an enhanced survival (up to 250%) compared to the parental strain, while the survival of the albino mutant was similar to that experienced by the parental strain; however, observed changes in survival were dose dependent. The cyclobutane pyrimidine dimers (CPDs), one of the major forms of DNA damage caused by UV exposure, appears as unrelated to the accumulation of carotenoids and cell survival. These results indicate that bearing higher torularhodin concentrations enhances UV-B survival in yeasts and, thus, the accumulation of this pigment constitutes an important mechanism that improves the resistance of yeasts to UV-B.

Seven strains representing a novel yeast species belonging to the genus Cryptococcus were isolated from different substrates from Patagonia, Argentina, and The Netherlands. Three strains were isolated from a meltwater river draining from the Frias glacier at Mount Tronador situated in Nahuel Huapi National Park (Patagonia) and four were isolated from apple surfaces in Randwijk, The Netherlands. Analysis of the D1/D2 large-subunit rRNA gene and ITS region sequences indicated that these strains represent a single species that is distinct from other species of the Tremellales clade. The name Cryptococcus spencermartinsiae sp. nov. is proposed to accommodate these strains. The type strain is CRUB 1230T (=CBS 10760T =DBVPG 8010T).

Background Strains of the astaxanthin producing yeast Xanthophyllomyces dendrorhous have been isolated from different cold regions around the earth, and the presence of double stranded RNA (dsRNA) elements was described in some isolates. This kind of viruses is widely distributed among yeasts and filamentous fungi and, although generally are cryptic in function, their studies have been a key factor in the knowledge of important fungi. In this work, the characterization and genetic relationships among dsRNA elements were determined in strains representatives of almost all regions of the earth where X. dendrorhous have been isolated. Results Almost all strains of X. dendrorhous analyzed carry one, two or four dsRNA elements, of molecular sizes in the range from 0.8 to 5.0 kb. Different dsRNA-patterns were observed in strains with different geographic origin, being L1 (5.0 kb) the common dsRNA element. By hybridization assays a high genomic polymorphism was observed among L1 dsRNAs of different X. dendrorhous strains. Contrary, hybridization was observed between L1 and L2 dsRNAs of strains from same or different regions, while the dsRNA elements of minor sizes (M, S1, and S2) present in several strains did not show hybridization with neither L1 or L2 dsRNAs. Along the growth curve of UCD 67-385 (harboring four dsRNAs) an increase of L2 relative to L1 dsRNA was observed, whiles the S1/L1 ratio remains constant, as well as the M/L1 ratio of Patagonian strain. Strains cured of S2 dsRNA were obtained by treatment with anisomycin, and comparison of its dsRNA contents with uncured strain, revealed an increase of L1 dsRNA while the L2 and S1 dsRNA remain unaltered. Conclusion The dsRNA elements of X. dendrorhous are highly variable in size and sequence, and the dsRNA pattern is specific to the geographic region of isolation. Each L1 and L2 dsRNA are viral elements able to self replicate and to coexist into a cell, and L1 and S2 dsRNAs elements could be part of a helper/satellite virus system in X. dendrorhous.

Seventy-one strains were isolated from the River Agrio–Lake Caviahue acidic aquatic environment in Argentina. Strains were isolated mainly from the most acidic section of the river (pH 1.8–2.7). According to the mini/microsatellite-primed PCR technique and physiological data, these strains are representatives of a single novel species of the genusCryptococcus. Analysis of the D1/D2 region of the large-subunit rRNA gene showed that the strains belong to the order Filobasidiales of the subphylum Agaricomycotina. The novel isolates formed a phylogenetic group withCryptococcus ibericus,Cryptococcus aciditoleransandCryptococcus metallitolerans, the most closely related species.This group, which is phylogenetically related to the Gastricus clade, is considered to be an ecoclade due to its peculiar ecology and physiology. The nameCryptococcus agrionensissp. nov. is proposed to accommodate these isolates, with strain CRUB 1317T(=CBS 10799T=JCM 15321T) as the type strain.

Here, we report on two novel sexual basidiomycetous red yeast species of the genus Cystofilobasidium. Cystofilobasidium lacus-mascardii sp. nov. is based on sexually compatible strains isolated from Lake Mascardi, an ultraoligotrophic lake in north-western Patagonia, Argentina. Following the discovery of the first isolate of this species, additional (sexually compatible) strains were isolated using a selective medium containing erythritol as the sole source of carbon. The second novel species corresponds to the sexual state of Cryptococcus macerans. In spite of accounts over the last 20 years of sexually compatible strains of this species, the complete life has never been observed. We provide evidence of a Cystofilobasidium-like basidial stage with teliospores and slender holobasidia, based on the study of self-fertile (homothallic) and self-sterile (heterothallic) isolates of Cryptococcus macerans. A revised molecular phylogeny of the genus Cystofilobasidium is presented and the most salient features of Cystofilobasidium lacus-mascardii sp. nov. (type strain CBS 10642(T) =PYCC 5819(T) =CRUB 1046(T)) and Cystofilobasidium macerans sp. nov. (type strain CBS 10757(T)) are discussed and compared with those of the remaining species in the genus. Information on additional Patagonian isolates belonging to the Cystofilobasidiales is also included in this report.

In this work, the photoprotective role of carotenoids in yeasts was analysed by contrasting the responses to UV-B of pigmented and naturally occurring albino strains of Sporobolomyces ruberrimus and Cystofilobasidium capitatum in different conditions. Albino and pigmented strains were confirmed to be conspecific by PCR fingerprinting and rDNA sequencing. Experimental exposure to UV-B conducted with both yeast species showed that the pigmented strains were more tolerant to UV-B than the albino strains and that the increment in carotenoid contents during the stationary growth phase enhance survivorship. These results indicated that carotenoid pigments afford UV-B protection in yeasts.

The Rio Agrio and Lake Caviahue system (RAC), in Northwestern Patagonia, is a natural acidic environment. The aims of this study were to characterize the yeast community and to provide the first ecological assessment of yeast diversity of this extreme aquatic environment. Yeast occurrence and diversity were studied at seven sites where the water pH varied between 1.8 and 6.7. Yeast CFU counts in the river ranged from 30 to 1200 CFU L(-1), but in the Lake the values were lower (30-60 CFU L(-1)). A total of 25 different yeast species were found, 11 of which belonged to undescribed taxa. Among these was an unusual strongly acidophilic Cryptococcus species. The RAC yeast community resembles that of acidic aquatic environments resulting from anthropic activities such as the São Domingos mines in Portugal and the Rio Tinto in Spain, respectively. The isolated yeast species were organized into different grades of adaptation to the RAC aquatic system. Based on the proposed grades, Rhodotorula mucilaginosa, Rhodosporidium toruloides and two novel Cryptococcus species were the most adapted species. These Cryptococcus species are apparently specialists of acidic aquatic environments, and might bear physiological features that possibly account for their ability to thrive in such extreme environments.

The yeast species Rhodotorula mucilaginosa is considered to be ubiquitous due to its world-wide distribution in terrestrial, freshwater and marine habitats, and to its ability to colonize a large variety of substrates. In this paper we assess the phenotypic and genetic variability of environmental isolates of R. mucilaginosa collected from natural and artificial environments in Patagonia, Argentina. A total of 97 strains were studied and sorted into three groups based on MSP-PCR fingerprinting results: A, which comprised 90% of the strains, including the type strain; and B and C which included 2 and 8% of the strains, respectively. The D1D2 sequencing did not differentiate any of the 3 groups, while ITS sequencing validated the existence of group C. This group was composed of Patagonian isolates of diverse origin. DNA-DNA reassociation studies confirmed the existence of significant genetic differences between group C and the type strain. In this study, which is the first on the intraspecific variability of a large set of R. mucilaginosa isolates, a considerable phenotypic and genetic heterogeneity was observed, however such differences were not enough to refute co-specificity. The study of Patagonian isolates allowed the detection of a genetically distinct group of R. mucilaginosa strains. (© 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

Eighteen yeast species of the genera Rhodotorula , Rhodosporidium , Sporobolomyces , and Sporidiobolus , each one represented by its type strain, were investigated with the objective of evaluating their carotenoid composition. The pigments were extracted from yeast cells, quantified by high pressure liquid chromatography diode array detector and the main compounds were confirmed by atmospheric pressure chemical ionization quadrupole mass spectrometry. Significant (P < 0.01) differences among several species and (or) genera were observed. Thirteen strains were seen to be able to produce carotenoids, from 16.4 to 184 μg/g cell dry mass and from 6.0 to 1993.4 μg/L culture. The main carotenoids produced were identified as torularhodin, torulene, γ-carotene, and β-carotene. The correlation matrix calculated on the basis of the carotenoid composition data matrix indicated significant (P < 0.01) relationships between torulene and torularhodin (r = 0.81), γ-carotene and torulene (r = 0.49), β-carotene and torulene (r = –0.72), as well as β-carotene and γ-carotene (r = 0.64). These significant correlation coefficients may suggest that species belonging to the genera Rhodosporidium, Sporobolomyces, and Sporidiobolus possess a carotenoid biosynthetic pathway analogous to that elsewhere postulated for Rhodotorula species.

As part of a project aimed at the selection of cold-adapted yeasts expressing biotechnologically interesting features, the extracellular enzymatic activity (EEA) of basidiomycetous yeasts isolated from glacial and subglacial waters of northwest Patagonia (Argentina) was investigated. Ninety-one basidiomycetous yeasts (belonging to the genera Cryptococcus , Leucosporidiella , Dioszegia , Mrakia , Rhodotorula , Rhodosporidium , Sporobolomyces , Sporidiobolus , Cystofilobasidium , and Udeniomyces ) were screened for extracellular amylolytic, proteolytic, lipolytic, esterasic, pectinolytic, chitinolytic, and cellulolytic activities. Over 15% of the strains exhibited three or more different EEAs at 4 °C and more than 63% had at least two EEAs at the same temperature. No chitinolytic or cellulolytic activities were detected at 4 and 20 °C. Cell-free supernatants exhibited significantly higher (P < 0.01) protease and lipase activities at ≤10 °C, or even at 4 °C. In light of these findings, cold environments of Patagonia (Argentina) may be considered a potential source of cold-adapted yeasts producing industrially relevant cold-active enzymes.

A novel population of the biotechnologically important yeast Xanthophyllomyces dendrorhous, the sexual stage of Phaffia rhodozyma, has been recently isolated for the first time in the southern Hemisphere (Patagonia, Argentina). The aim of the present work was to phenotypically and genotypically characterize two representative strains of this new population, and assess such strains as a potential biotechnological source of astaxanthin, fatty acids and extracellular enzymes. Minor variations were found in physiological tests. PCR fingerprinting studies (MSP-PCR) showed the main differences between X. dendrorhous Patagonian and Type strains. Patagonian strains accumulated a xanthophyll-like pigment, which was identified as astaxanthin. These strains showed low fatty acids content (mainly polyunsaturated fatty acids) and, of a total of six extracellular enzymes tested, only produced amylase. Genetic differences between Patagonian and collection X. dendrorhous strains could be explained by geographic isolation and habitat specificity.

Phaffia rhodozyma (sexual form, Xanthophyllomyces dendrorhous ) is a basidiomycetous yeast that has been found in tree exudates in the Northern Hemisphere at high altitudes and latitudes. This yeast produces astaxanthin, a carotenoid pigment with biotechnological importance because it is used in aquaculture for fish pigmentation. We isolated X. dendrorhous from the Southern Hemisphere (Patagonia, Argentina), where it was associated with fruiting bodies of Cyttaria hariotii , an ascomycetous parasite of Nothofagus trees. We compared internal transcribed spacer (ITS)-based phylogenies of P. rhodozyma and its tree host (Betulaceae, Corneaceae, Fagaceae, and Nothofagaceae) and found them to be generally concordant, suggesting that different yeast lineages colonize different trees and providing an explanation for the phylogenetic distance observed between the type strains of P. rhodozyma and X. dendrorhous . We hypothesize that the association of Xanthophyllomyces with Cyttaria derives from a previous association of the yeast with Nothofagus , and the sister relationship between Nothofagaceae and Betulaceae plus Fagaceae correlates with the phylogeny of X. dendrorhous strains originating from these three plant families. The two most basal strains of X. dendrorhous are those isolated from Cornus , an ancestral genus in the phylogenetic analysis of the host trees. Thus, we question previous conclusions that P. rhodozyma and X. dendrorhous represent different species since the polymorphisms detected in the ITS and intergenic spacer sequences can be attributed to intraspecific variation associated with host specificity. Our study provides a deeper understanding of Phaffia biogeography, ecology, and molecular phylogeny. Such knowledge is essential for the comprehension of many aspects of the biology of this organism and will facilitate the study of astaxanthin production within an evolutionary and ecological framework.

In this paper we present the results of research on the occurrence, induction and role of photoprotective compounds (PPCs) present in native aquatic yeasts from freshwater Patagonian ecosystems. We focus on the effect of UV radiation (UVR) as a factor that controls the level of photoprotection of yeasts, and explore its potential significance in shaping yeast distributional patterns. The research presented here combines field surveys and laboratory work, including the isolation and culture of native yeasts strains, and laboratory assays under different radiation conditions. The results obtained suggest that yeasts are common dwellers of oligotrophic Patagonian water bodies, and provide the first evidence of the distribution of PPC (carotenoid and mycosporine)-producing yeasts in temperate freshwaters. A greater proportion of carotenogenic yeasts were observed in high-elevation lakes. The yeast strains isolated from these environments were found to produce higher amounts of mycosporines (MYCs), and to present higher tolerance to UVB exposure than those from piedmont lakes. Patagonian yeasts have only one type of MYC, mycosporine-glutaminol-glucoside (myc-glu-glu), which seems common to all other yeasts. By analyzing the production of myc-glu-glu in a large number of yeasts belonging to different taxonomic groups, we propose that this compound may have potential use as a chemotaxonomic marker in yeast systematics. Collectively, our work reveals that in Patagonian freshwater yeasts there is an apparent relationship between the ability to produce PPCs, their tolerance to UV exposure and their success in colonizing habitats highly exposed to UVR.

Mycosporine-like amino-acids (MAAs) are found in aquatic bacteria, algae, and animals. A related compound, the mycosporine-glutaminol-glucoside (myc-glu-glu), has recently been reported in freshwater yeasts. Although animals depend on other organisms as their source of MAAs, they can efficiently accumulate them in their tissues. In this work we assessed the potential transfer of the yeast mycosporine myc-glu-glu from the diet into the copepod Boeckella antiqua and the ciliate Paramecium bursaria. For this purpose, we performed experiments to study the feeding of B. antiqua and P. bursaria on the yeast Rhodotorula minuta and their ability to bioaccumulate myc-glu-glu. Bioaccumulation of myc-glu-glu in B. antiqua was assessed through long-term factorial experiments manipulating the diet (Chlamydomonas reinhardii and C. reinhardii + yeasts) and radiation exposure (PAR and PAR + UVR). Shorter term experiments were designed in the case of P. bursaria. The composition and concentration of MAAs in the diet and in the consumers were determined by HPLC analyses. Our results showed that even though both consumers ingested yeast cells, they were unable to accumulate myc-glu-glu. Moreover, when exposed to conditions that stimulated the accumulation of photoprotective compounds (i.e. UVR exposure), an increase in MAAs concentration occurred in copepods fed C. reinhardii plus yeasts as well as in those fed only C. reinhardii. This suggests that the copepods were able to modify their tissue concentrations of MAAs in response to environmental clues but also that the contribution of yeast mycosporines to total MAAs concentration was negligible.

The ability to produce mycosporines (MYCs) in 157 pigmented yeast strains (eight genera, 25 species) isolated from natural environments of Patagonia (Argentina) was assessed. The strains belong to four taxonomic groups: the Sporidiobolales and Erythrobasidium clade of the class Urediniomycetes, and Cystofilobasidiales and Tremellales of the class Hymenomycetes. Induction of MYCs did not occur in all yeast strains tested and appeared to be an exclusive trait of members of the Erythrobasidium clade and Tremellales. This is the first report on the production of MYCs by pigmented species from the latter group, as well as the first extensive screening of mycosporinogenic yeasts. The consistent occurrence of MYCs in some specific phylogenetic groups suggests this trait bears evolutionary significance and that the presence/absence of MYCs may have potential applications in yeast systematics.

During a survey of carotenogenic yeasts carried out in north-western Patagonia (Argentina), several ballistoconidia-producing strains belonging to the order Sporidiobolales were isolated from aquatic environments. Five strains were found to represent two novel species, for which the names Sporidiobolus longiusculus and Sporobolomyces patagonicus are proposed, with CBS 9654T (=PYCC 5818T=CRUB 1044T) and CBS 9657T (=PYCC 5817T=CRUB 1038T) as the type strains, respectively. The elongated basidia, which are five to six times longer that those of the remaining species of the genus Sporidiobolus, are a particular micromorphological feature of Sporidiobolus longiusculus. On the basis of the sequences of the D1/D2 domains of the 26S rRNA gene, the species most closely related to Sporidiobolus longiusculus is Sporobolomyces bannaensis, whereas Sporobolomyces marcillae is the closest relative of Sporobolomyces patagonicus. Complete internal transcribed spacer sequence analysis confirmed the separate position of Sporidiobolus longiusculus, whereas for Sporobolomyces patagonicus no nucleotide differences were found with respect to Sporidiobolus pararoseus CBS 491T. Negative mating experiments between strains of Sporobolomyces patagonicus and strains of Sporidiobolus pararoseus together with the low DNA–DNA reassociation values for the type strains of the two species validated the proposal of Sporobolomyces patagonicus as a distinct species. Information on additional Patagonian Sporobolomyces isolates is also included in this report.

High-performance liquid chromatography (HPLC), alone or in combination with mass spectrometry (MS), was used to screen the basidiomycetous yeasts Rhodotorula minuta and R. slooffiae isolated from lakes for the presence of UV-absorbing compounds. Mycosporine-glutaminol-glucoside (maximum absorption, 310 nm), a UV-photoprotective mycosporine known in terrestrial fungi, was the major UVabsorbing compound found in these species. This is the first identification of a mycosporine in yeasts. The presence of this compound seems to be a promising chemotaxonomical marker for yeast systematics. Copyright  2004 John Wiley & Sons, Ltd.

Twelve yeasts isolated from lakes of Northwestern Patagonia, Argentina, belonging to eight genera (Sporobolomyces, Sporidiobolus, Rhodotorula, Rhodosporidium, Cystofilobasidium, Cryptococcus, Torulaspora, and Candida) were analysed for their ability to produce photoprotective compounds. For this purpose, three laboratory experiments were performed to study the effect of photosynthetically active radiation (PAR) and PAR in combination with UV radiation (PAR + UVR) on the production of carotenoids and mycosporines. The synthesis of carotenoid compounds was clearly stimulated in six out of nine red yeast strains tested upon exposure to PAR or PAR + UVR; however, the latter conditions produced a stronger response than PAR alone. The increase in carotenoids in the red strains under PAR + UVR irradiation showed a negative exponential relationship with their basal carotenoid content, suggesting that cells with higher constitutive levels of carotenoids are less responsive to induction by PAR + UVR. Three red yeasts, Rhodotorula minuta, Rh. pinicola, and Rhodotorula sp., and the colourless Cryptococcus laurentii produced a UV-absorbing compound when exposed to PAR or PAR + UVR. This compound showed an absorption maximum at 309–310 nm and was identified as mycosporine-glutaminol-glucoside (myc–glu–glu). In these strains, exposure to PAR or PAR + UVR resulted in elevated concentrations of both carotenoids and myc–glu–glu. This is the first report on the production of mycosporines by yeasts. All strains that developed under PAR + UVR were able to synthesise carotenoids either constitutively or in response to PAR exposure, and a few of them also produced myc–glu–glu when exposed to PAR. Collectively, our results suggest that the presence of carotenoids, either alone or in combination with mycosporines, are required for sustaining growth under exposure to PAR + UVR in the freshwater yeast strains studied.

Recientemente, se ha encontrado que la distribución natural, el hábitat y la diversidad genética de levaduras productoras de astaxantina (p. ej., Phaffia rhodozyma, sinónimo Xanthophyllomyces dendrorhous) son mucho mayores de lo que se pensaba. P. rhodozyma se explota biotecnológicamente debido a su capacidad para producir el pigmento carotenoide astaxantina y, por lo tanto, se utiliza como una fuente natural de este pigmento para la acuicultura. También se encontró que esta levadura es capaz de sintetizar el potente protector solar UVB micosporina-glutaminol-glucósido (MGG). Por lo tanto, más estudios ambientales para dilucidar sus aspectos ecológicos y detectar nuevas cepas potenciales productoras de astaxantina y MGG son necesarios. Sin embargo, la obtención de nuevos aislamientos de P. rhodozyma y especies relacionadas no siempre es fácil debido a su baja abundancia y a la presencia de otras levaduras simpátricas y pigmentadas. En este trabajo se describe el desarrollo exitoso de un cebador especie-específico que tiene la capacidad de detectar rápidamente y con precisión cepas representativas de todos los linajes del género Phaffia previamente reportados (incluyendo nuevas especies del género) y excluir especies estrechamente relacionadas. Para ello, se diseñó un cebador de 20 nucleótidos (denominado PhR) para ser utilizado en combinación con los cebadores universales ITS3 y NL4 en una amplificación multiplex. El método propuesto tiene la sensibilidad y la especificidad requerida para la detección precisa de nuevos aislamientos y, por lo tanto, representa una importante herramienta para la búsqueda ambiental de nuevas levaduras productoras de astaxantina. It has been recently found that the natural distribution, habitat, and genetic diversity of astaxanthin-producing yeasts (i.e. Phaffia rhodozyma, synonym Xanthophyllomyces dendrorhous) is much greater than previously thought. P. rhodozyma is biotechnologically exploited due to its ability to produce the carotenoid pigment astaxanthin and thus, it is used as a natural source of this pigment for aquaculture. P. rhodozyma was also capable of synthesizing the potent UVB sunscreen mycosporine-glutaminol-glucoside (MGG). Therefore, further environmental studies are needed to elucidate its ecological aspects and detect new potential strains for the production of astaxanthin and MGG. However, obtaining new isolates of P. rhodozyma and related species is not always easy due to its low abundance and the presence of other sympatric and pigmented yeasts. In this work we report a successful development of a species specific primer which has the ability to quickly and accurately detecting isolates representing all known lineages of the genus Phaffia (including novel species of the genus) and excluding closely related taxa. For this purpose, a primer of 20 nucleotides (called PhR) was designed to be used in combination with universal primers ITS3 and NL4 in a multiplex amplification. The proposed method has the sensitivity and specificity required for the precise detection of new isolates, and therefore represents an important tool for the environmental search for novel astaxanthin-producing yeasts. Fil: Colabella, Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones en Biodiversidad y Medioambiente. Universidad Nacional del Comahue. Centro Regional Universidad Bariloche. Instituto de Investigaciones en Biodiversidad y Medioambiente; Argentina Fil: Libkind Frati, Diego. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones en Biodiversidad y Medioambiente. Universidad Nacional del Comahue. Centro Regional Universidad Bariloche. Instituto de Investigaciones en Biodiversidad y Medioambiente; Argentina

Fungos endofíticos são aqueles que, pelo menos durante parte do ciclo de vida, habitam tecidos vegetais vivos sem causar sintomas aparentes de doença ou efeitos negativos ao hospedeiro. Nos últimos anos, plantas com histórico etnobotânico vêm sendo alvos constantes de estudos de bioprospecção, pois acredita-se que algumas propriedades medicinais atribuídas ao vegetal também possam estar relacionadas à produção de metabólitos secundários por seus fungos endofíticos. O primeiro capítulo deste trabalho teve o objetivo de caracterizar a diversidade de fungos endofíticos associados às plantas medicinais Myrciaria floribunda, Eugenia aff. neomyrtifolia e Alchornea castaneifolia e avaliar seu potencial quanto à produção de metabólitos bioativos. Um total de 93 fungos filamentosos foram isolados e identificados por meio do sequenciamento da região espaçadora transcrita interna (ITS) do rRNA. Trinta e oito extratos fúngicos apresentaram atividade contra pelo menos um dos micro-organismos alvo avaliados. Estudos sobre a biogeografia de micro-organismos são escassos e para elucidar se os padrões de diversidade endofítica são correlacionados à filogenia dos seus hospedeiros ou são influenciados pelas variáveis ambientais ou distância foi avaliada a distribuição da comunidade de fungos endofíticos associados à hospedeiros filogeneticamente relacionados de plantas da tribo Myrtae. O estudo foi realizado seguindo-se três escalas espaciais: regional, 101-5.000 km; local, 0-100 km e microescala, 0-1 km. Um total de 960 isolados foi obtido, agrupados e identificados em 52 espécies distintas. A similaridade da comunidade de fungos endofíticos diminuiu com o aumento da distância geográfica, o que foi observado pela análise de decaimento da distância. A análise de escalonamento metrico não dimensional (NMDS) mostrou que os fungos endofíticos agruparam-se a um nível de escala espacial local. Além disso, a análise de regressão múltipla de matrizes (MRM) sugeriu que a similaridade da comunidade endofítica depende da escala espacial analisada. Em outro trabalho realizado foi mostrado que os fungos endofíticos também podem interagir com fungos micorrízicos arbusculares melhorando a adapatabilidade das plantas por meio da promoção do crescimento vegetal. Neste trabalho, cento e cinquenta isolados de fungos endofíticos foram isolados das raízes de M. communis e oito diferentes grupos foram selecionados para o estudo com Sorghum vulgare. Os fungos não foram capazes de colonizar as raízes de sorgo, porém apresentaram efeitos significativos no crescimento e colonização por micorrizas arbusculares. A quarta parte deste trabalho teve como finalidade avaliar a diversidade de fungos endofíticos associados à Myrtus communis utilizando-se técnicas tradicionais de isolamento e por meio de eletrofose em gel de gradiente desnaturante (DGGE). Um total de 113 isolados de fungos endofíticos foi obtido pela técnica de isolamento em meio de cultura em ágar batata dextrosado (BDA). A similaridade da comunidade endofítica não diminuiu com o aumento da distância geográfica. Os resultados do presente trabalho sugerem uma elevada diversidade de fungos endofíticos associados a hospedeiros presentes em diversos ecossistemas. Além disso, nossos resultados também mostraram o potencial biotecnológico destes fungos como produtores de substâncias antimicrobianas e como promotores do crescimento vegetal. Endophytic fungi inhabit healthy plant tissues during at least one stage of their life cycle without causing any apparent symptom of disease or negative effects on the hosts. In the last years, the endophytic fungal mycota associated with popular medicinal plants have been perfomed because some medicinal properties have been associated to secondary metabolities production by their endophytic fungi associated. Fungal endophyte communities associated with leaves of Myrciaria floribunda, Alchornea castaneifolia, and Eugenia aff. bimarginata were examined, collected from Brazilian Cerrado ecosystems, and studied for their ability to produce antimicrobial activity. A total of 93 isolates of endophytic fungi were obtained and identified by sequencing of internal transcribed spacer (ITS) regions of the rRNA gene. Thirty-eight fungal extracts presented antimicrobial activity against at least one of the different target microorganisms tested. The biogeography of microorganisms is poorly understood and to to address whether patterns of endophyte diversity are correlated with host phylogeny, environmental variables and distance we designed a geographic survey of related host tree of Myrtae tribe. The study was performed at the following spatial scales: regional, 101-5,000 km; local: 0-100 km and microscale: 0-1 km. A total of 960 isolates of endophytic fungi were obtained from 3,000 leaf fragment samples. These isolates were grouped and identified into 52 species. We found that the similarity of endophytic fungal communities decreases with increasing geographical distance, showed by the distance decay analysis. The nonmetric multidimensional scaling (NMDS) analysis showed that the fungal endophytes communities were grouped at a local scale. The multiple regression on matrices (MRM) suggested that the fungal endophytic community similarity depends on the scale sampled used. Other study showed that endophytic fungi can interact with arbuscular micorryzal, increasing the growth plant. A total of 150 isolates of endophytic fungi were obtained from roots of Myrtus communis and seven groups were selected to interactions study with Sorghum vulgare. None of the root endophytic fungi tested colonized the root of sorghum, but significantly increased the growth and arbuscular mycorrhizal colonization of sorghum. The fourth part of this work evaluated the endophytic community diversity of Myrtus communis by classical isolation from leaves fragments and denaturing gel gradient electrophoresis (DGGE). A total of 113 fungal isolates were obtained and identified by classical isolation in potato dextrose agar. It was found that the similarity of endophytic fungal communities did not decrease with increasing geographical distance. The results obtained in this work suggest a high endophytic fungal diversity associated with Myrtaceae host tree presents indifferent ecossistems. Moreover, these micro-organismos represents a potencial source of substances that can be used to development of new antimicrobial drugs and as a growth promoting fungi.