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A study was performed on a collection of 84 isolates from decaying plant tissues and soils in Argentina previously identified as Trichoderma harzianum. Based on multiple phenotypic characters and multilocus phylogenetic analyses, 10 species were distinguished, three of which are described as new species: T. austroindianum, T. hortense, and T. syagri. Among the remaining seven identified species, the following five can be added to the Argentine mycobiota: T. afarasin, T. afroharzianum, T. endophyticum, T. guizhouense, and T. neotropicale. Trichoderma afroharzianum and T. endophyticum were the most frequent species found in the samples. In addition, a collection of isolates previously identified as T. harzianum with antagonistic abilities were reidentified as T. afroharzianum, thus highlighting the importance of correct identification of biocontrol species. Instituto Nacional de Tecnología Agropecuaria/[2019-PD-E4-I069-001]/INTA/Argentina Agencia Nacional de Promoción de la Investigación, el Desarrollo Tecnológico y la Innovación/[PICT2018-4436]/ANPCyT/Argentina UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigaciones en Productos Naturales (CIPRONA)

Improving micropropagation success rate is important since this technique allows the development of other biotechnological tools, such as genetic transformation, in vitro polyploidization, or endophyte inoculation. This is particularly relevant for grasses, as they are traditionally considered recalcitrant to plant tissue culture. Grasses of the subfamily Pooideae establish mutualistic associations with grass endophyte fungi of the genus Epichloë. These associations are specific, as each host species is associated with one or few Epichloë species. Lolium arundinaceum (Schreb.) Darbysh. is a perennial C3 species, native to Europe, which has become an important cool season forage crop worldwide. This grass usually establishes beneficial symbioses with E. coenophiala, but these benefits depend on both plant and fungal genetics and environmental conditions. Micropropagation protocols have been published for L. arundinaceum, but the influence of Epichloë endophytes was never examined. The aim of this manuscript was to study the effect of E. coenophiala on different micropropagation stages of L. arundinaceum and growth of the regenerated plantlets. Association with E. coenophiala (E+) increased callus induction, enhanced their proliferation, and increased shoot regeneration in L. arundinaceum. However, the biomass of plantlets regenerated from E+ seeds was lower than from non-associated seeds, both in vitro and ex vitro, and the in vitro multiplication rate also decreased. Contrary to other endophyte Epichloë-host associations with Lolium multiflorum and Bromus auleticus, the association with E. coenophiala did not improve tall fescue micropropagation. We can conclude that the effects of Epichloë endophytes on micropropagation and growth of their host differ for each endophyte-host association. Fil: Regalado González, Jose Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Berdion Gabarain, Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Tossi, Vanesa Eleonora. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Iannone, Leopoldo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Pitta Alvarez, Sandra Irene. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Novas, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina

Bromus auleticus (Trin.) is a grass native to the southern cone with important agronomical potential as fodder. Different breeding programs have been initiated with this grass, but plant tissue culture techniques could not be used because B. auleticus is recalcitrant. The aim of the present study was to develop a micropropagation protocol in the genus Bromus and to investigate if the association between B. auleticus and Epichloë endophytes affected in vitro culture and growth of micropropagated plantlets. In different micropropagation stages, better results were obtained with endophyte-infected (E+) seeds compared to endophyte-free (E−) seeds. The E+ seeds presented higher percentages of in vitro germination (82 ± 5 vs. 57 ± 6%), callus induction (72 ± 6 vs. 37 ± 6%), and plant regeneration from callus (89 ± 5 vs. 13 ± 5%). We also compared the biomass of shoot complexes and regenerated plantlets. After 4 weeks of culture, shoot complexes obtained from E+ seeds reached greater weight than the ones regenerated from E− seeds (173 ± 24 vs. 74 ± 9 mg). More than the 80% of the regenerated shoot complexes were rooted ex vitro and acclimated, regardless of their origin (E+ or E−). Finally, after 4 weeks of acclimatization, the plantlets regenerated from E+ seeds reached a greater weight than the ones from E− seeds, (461 ± 64 vs. 172 ± 25 mg). These results indicate that the use of endophyte-infected (E+) seeds enhances significantly B. auleticus micropropagation and promotes growth of the regenerated plantlets. Fil: Regalado González, Jose Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Berdion Gabarain, Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Vignale, Maria Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Novas, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Pitta Alvarez, Sandra Irene. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Iannone, Leopoldo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina

Some grasses are co-infected by ‘castrator’ fungal pathogens, such as Ustilago bullata, and beneficial seed-transmitted Epichloë endophytes that compete for plant photosynthates and flowers. Epichloë-infected (E+) and Epichloë-free (E−) seeds of Bromus auleticus were inoculated (S+) or not (S−) with teliospores of U. bullata and an experiment under field conditions was performed to evaluate the effect of these fungal infections on different host plant traits. Head smut disease incidence was almost 0% in E+ treatment whereas in endophyte-free plants the disease incidence reached 33%, a result in concordance with the lower number of seeds produced by E−S+ plants. Neither the smut nor the endophyte affected seedling emergence or growth. Plant survival was significantly decreased by the smut in E− plants and was associated with a higher biomass production in E+ treatments. Our results provide new insights about the ecology of Epichloë endophytes and their protective and growth-promoting role in the interaction of its host with U. bullata. Fil: Iannone, Leopoldo Javier. Universidad de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Vignale, Maria Victoria. Universidad de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Pinget, Albertina Daniela. Universidad de Concepción del Uruguay; Argentina Fil: Re, A.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina Fil: Mc Cargo, P. D.. Universidad de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Novas, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina. Universidad de Buenos Aires; Argentina

Italian ryegrass (Lolium multiflorum) is an annual grass considered as one of the most important temperate forage grasses in the world. However, it is recalcitrant to plant tissue culture techniques hindering its genetic manipulation. Epichloë occultans is an endophytic fungus associated with L. multiflorum. This symbiosis causes improvements in physiological and ecological traits of the host plants. The objective of this work was to study the effect of E. occultans on L. multiflorum micropropagation. We compared the response of endophyte-infected (E+) and endophyte-free (E−) seeds in different micropropagation stages. The E+ seeds were more successful than E− seeds in in vitro germination (83 ± 5 vs. 63 ± 6%), callus induction (78 ± 5 vs. 57 ± 6%), callus proliferation (average diameter of 21.5 ± 1.3 mm in two subcultures vs 17.3 ± 0.8 mm in three subcultures) and plant regeneration from callus (83 ± 7 vs. 30 ± 8%). These results indicate that E. occultans enhances significantly L. multiflorum micropropagation. The use of endophyte-infected (E+) seeds can be a solution to make this grass more amenable to different biotechnological tools, such as the genetic transformation. Fil: Regalado González, Jose Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Vignale, Maria Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Novas, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Pitta Alvarez, Sandra Irene. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Iannone, Leopoldo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina

Background and aims: We studied, through exudates employment, the effect of Epichloë (endophytic fungi), both independently and in association with Bromus auleticus (grass), on arbuscular mycorrhizal fungi (AMF) colonization, host and neighbouring plants biomass production and soil changes. Methods: Through in vitro and greenhouse experiments, Epichloë endophytes effect on AMF development was evaluated. In vitro studies of exudates effect on Gigaspora rosea and Rhizophagus intraradices were performed using root or endophyte exudates. A 6-month greenhouse experiment was conducted to determine Bromus auleticus endophytic status effect and endophyte exudates role in biomass production, neighbouring plants mycorrhizal colonization and soil properties. Results: Endophyte exudates and E+ plant root exudates promoted in vitro AMF development in the pre-infective stage of G. rosea and in carrot root culture mycelium of R. intraradices in a dose-response relationship, while control media and E- plants exudates had no effect. R. intraradices colonization and plant growth was clearly increased by endophytes and their exudates. Conclusions: This is the first work evidencing the direct effect of Epichloë endophytes and infected plants root exudates on AMF extramatrical development. While higher levels of AMF colonization were observed in E+ plants, no clear effect was detected in neighbouring plants colonization, plant biomass or soil properties. Fil: Vignale, Maria Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Iannone, Leopoldo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Scervino, Jose Martin. 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: Novas, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina

This study compares the culturable fungal endophytic community inhabiting leaves of Epichloë-infected (E+) and Epichloë-free (E-) plants of Poa bonariensis. Plants were collected from a wild population (dioecious species) where E+ and E- plants co-exist. Fungal endophytes were isolated and then morphology and ribosomal DNA internal transcribed spacer sequences were used for the identification of isolates. In total 24 different taxa of non-Epichloë (NE) endophytes were identified. No significant differences were found in richness and diversity indexes of NE-endophytes between E+ and E- plants, or between pistillate (P) and staminate (S) plants. When we analyzed these effects based on the count of the most abundant genera, Alternaria and Stemphylium infection frequencies decreased in S plants, while in E+ plants, this tendency was observed for Alternaria. Our results suggest that Epichloë and the sex of the host plant may modulate some of the dominant foliar endophytic fungal taxa. Fil: Mc Cargo, Patricia Débora. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Iannone, Leopoldo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Soria, Marcelo. Universidad de Buenos Aires. Facultad de Agronomía; Argentina Fil: Novas, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina

Carob flour is obtained from pods of some species of Prosopis, leguminous trees that abound in many desert habitats worldwide. Currently, this product is available in healthy food stores in several countries, including Argentina, as a nontraditional meal of growing interest with multiple applications for the preparation of puddings, biscuits and snacks, among others. The objective of the present study was to evaluate the microbial quality of carob flour on basis of the presence of deteriorative and pathogenic microorganisms. Fungal diversity of the mycobiota was also studied with a special interest in toxigenic fungi. Eighteen samples of carob flour (Prosopis flexuosa) were analysed. Standard plate count of aerobic mesophilic bacteria showed levels of contamination ranging from

A field experiment was performed to study the effect of Epichloë endophytes of the wild forage grass Bromus auleticus on mycorrhizal colonization, seedling emergence and growth parameters in three heterospecific grasses of agronomic interest. Plots with B. auleticus associated with Epichloë (E+) and non-associated (E-) plants were established and 3 y later, seeds of Lolium multiflorum (E+ or E-), Schedonorus arundinaceus (E+ or E-) and Bromus catharticus (not establishing associations with Epichloë) were sown as B. auleticus neighbors. B. auleticus E+ plants induced a significant increase in mycorrhizal colonization of the neighbor grasses, even higher in the E- neighbor plants (L. multiflorum and S. arundinaceus), in comparison to B. auleticus E- neighbors. Seedling emergence, root length, leaves number or shoot dry weight were not affected. This is the first approach to studying these triple interactions (endophyte-grass-mycorrhiza) involving wild and forage grasses in the field. Our results show that the effect of Epichloë extends further than its own host and suggest an interaction among endophytes of heterospecific neighbors at the rhizosphere level.Keywords arbuscular mycorrhizal fungi, Bromus auleticus, Lolium multiflorum, multiple mutualistic symbiosis, root exudates, Schedonorus arundinaceus Fil: Vignale, Maria Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Iannone, Leopoldo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Novas, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina

Cattle breeding is an important economical activity in Argentina, highly dependent on grass production. In the last decades, grasslands zones were reduced and confined to less productive lands due to the advance of agronomical cultures. Therefore, it is important to develop new strategies to improve forage production. New eco-friendly trends in plant growth promotion include the use of microbial endophytes, but the in vitro studies of plant-bioinoculant interactions is limited by the scarce current technological development. In this work, we use a micropropagation protocol for Lolium multiflorum, developed in a previous work, to study the effect of bacterization with actinobacterial endophytes, isolated from Argentine native grasses, on the growth of L. multiflorum in vitro plantlets. To achieve this objective, L. multiflorum plantlets were inoculated with three Micromonospora strains (SB3, TW2.1 and TW2.2). The results obtained showed that the effect of actinobacterial inoculation depends on the Micromonospora strain used. The inoculation with SB3 promoted plant growth, increasing plant biomass, root length and the rate of plantlets ready to be acclimatized after 4 weeks of in vitro culture. Strain TW2.1 did not show, statistically, differences compared to control treatments, while TW2.2 inhibited plant growth, decreasing plant biomass, root length and the rate of plants ready to acclimatize. Our results showed that Micromonospora strain SB3 could be a good candidate to use in breeding programs for L. multiflorum and other grasses to increase their yield. Fil: Della Mónica, Ivana Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Novas, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Iannone, Leopoldo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Querejeta, Giselle Andrea. Universidad Nacional de General Sarmiento. Instituto de Ciencias. Área de Química; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Scervino, Jose Martin. 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: Pitta Alvarez, Sandra Irene. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental. Laboratorio de Cultivo Experimental de Microalgas; Argentina Fil: Regalado González, Jose Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental. Laboratorio de Cultivo Experimental de Microalgas; Argentina

Asexual Epichloë endophytes establish mutualistic symbioses with grasses, improve fitness of their hosts and modify the surrounding environment. To test the hypothesis that this symbiotic association increases the abundance and diversity of phosphate-solubilizing fungi (PSF), a pot experiment was conducted combining two endophytic statuses: Epichloë-infected (E+) and non-infected (E−) Bromus auleticus plants, and two soil types collected from agricultural (A) and non-agricultural (NA) fields. Soil fungi were isolated at the beginning of the experiment and 12 months after the introduction of B. auleticus, and tested for their inorganic P (Pi)-solubilizing capability. Arbuscular mycorrhizal colonization in B. auleticus roots of E+ and E− plants was also analyzed. PSF abundance was affected by the endophytic status and by the type of soil; the highest value was detected in the E−NA treatment, followed by the E+A treatment. PSF diversity was higher in NA than in A soils and higher in soils treated with E+ than in those treated with E−. Arbuscular mycorrhizal fungi colonization was higher in E+ plants. We hypothesize that the positive association between Epichloë endophytes and mycorrhizal fungi with an increase in the PSF diversity would generate an increase in the phosphorus (P) available to plants. Fil: Arrieta, Alejandro Matias. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Iannone, Leopoldo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Scervino, Jose Martin. 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: Vignale, Maria Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Novas, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina

In this work we performed morphological and molecular phylogenetic analyses (based on sequences of calmodulin M [calM], translationelongation factor 1-α [tefA] and b-tubulin [tubB] genes) to characterize the diversity of Epichloë endophytes in Bromus setifolius and Phleum alpinum. The phylogenies obtained from the three genes were congruent and allowed differentiation of three lineages of endophytes that also presented morphological differences. One lineage corresponds to the previously described species Epichloë tembladerae, which is present in a wide range of native grasses from Argentina including B. setifolius and P. alpinum. Another genotype isolated only from B. setifolius is a non-hybrid endophyte, a rare condition for the South American Epichloë endophytes. Isolates of this genotype, described herein as a new variety, Epichloë typhina var. aonikenkana, presented waxy colonies at maturity and a low production of conidia. The third lineage, exclusively found in isolates from P. alpinum, is a hybrid between E. typhina and a common ancestor of E. amarillans and E. baconii. Isolates of this lineage produce abundant conidia that are variable in shape and size. Based on its unique phylogenetic position and morphology, we propose the new species, Epichloë cabralii for this lineage. The new combinations Epichloë tembladerae and E. pampeana also are proposed for the previously described Neotyphodium tembladerae and Neotyphodium pampeanum species. Fil: Mc Cargo, Patricia Débora. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Iannone, Leopoldo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Vignale, Maria Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Schardl, Christopher L.. University Of Kentucky; Estados Unidos Fil: Rossi, Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentina

In this work it was studied for the first time whether asexual Epichloë (Neotyphodium) endophytes of Bromus auleticus, protect their host plants against the pathogenic fungus Ustilago bullata.Seeds of two different ecotypes of B. auleticus, one of them infected with the endophyte Neotyphodium pampeanum (NpE+) and the other infected with the endophyte Neotyphodium tembladerae (NtE+) and their respectively endophyte-free (NpE-/NtE-) counterparts were used. Seeds of each ecotype and endophytic status were superficially disinfected and were randomly assigned to different treatments named: S+ (smut fungus inoculated) and S- (mock-inoculated). It was evaluated the effect of U. bullata infection on plant characteristics in every stage of their life cycle: seedling emergence, vegetative growth, mortality and smut symptoms in the florets.In NtE+ infected plants, smut disease was almost completely suppressed, whereas in their endophyte-free counterparts (NpE-) the incidence of smut symptoms reached 64%. In NpE+ infected plants smut incidence was significantly lower (7%) than in endophyte-free plants (39%). Although U. bullata infection decreased the emergence rate of both endophyte-infected and endophyte-free plants, neutral or protective effects of the endophytes were observed in seedling development and survival. The survival during the first year of NtE+ plants was higher than in their NtE- counterparts.These results indicate a strong beneficial effect of vertically transmitted endophytes against this pathogen. Fil: Vignale, Maria Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Astiz Gassó, Marta Mónica. Universidad Nacional de La Plata. Facultad de Ciencias Agrarias y Forestales. Instituto Fitotécnico de "Santa Catalina"; Argentina Fil: Novas, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Iannone, Leopoldo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina

Asexual, vertically transmitted fungal endophytes of thegenus Neotyphodium are considered to enhance growth,stress resistance and competitiveness of agronomicgrasses, but have been suggested to have neutral ordeleterious effects on wild grasses. We studied whetherthe associations between Bromus auleticus and theirendophytes,NeotyphodiumpampeanumandNeotyphodiumtembladerae,arebeneficialforthiswildforagegrassnativeto South America. In a greenhouse experiment,3-month-old endophyte-infected plants (E+) showedenhanced growth relative to their endophyte-free coun-terparts(E)).Inaone-year-longexperimentinthefield,E+plantsshowedhighersurvivalandregrowthrate,andproduced more biomass and seeds than E) plants. OnlywithrespecttotheseedoutputdidN. tembladeraeseemtobedetrimental,whereasN. pampeanumdidnotaffectthisparameter. No differences were observed with respect tothe germination of seeds produced by E+ or E) plants.LolinealkaloidsweredetectedinN. pampeanum-infectedplants. Our results show that, similar to results withagronomic grasses, in wild grasses, the symbiosis withNeotyphodium species could be mutualistic.Keywords: endophytes, wild grasses, Bromus, growth,reproduction

Neotyphodium endophytes and arbuscular mycorrhizal (AM) fungi are common constituents of natural grasslands. The plant?endophyte symbiosis can introduce changes in soil conditions that affect the density and activity of different functional groups of soil organisms. A few experiments have reported that leaf endophytes can reduce mycorrhizal sporulation and colonization of host roots in agronomic hosts, contrasting with the studies we have conducted in native grasses from Argentina. In the present work we performed in vitro assays to evaluate the effect of root and endophyte exudates on the pre-infective state of mycorrhizal fungi (Gigaspora margarita and G. rosea). Plant roots of Bromus setifolius, from populations of Patagonia and four strains of Neotyphodium were used to obtain the exudates. Root exudates of infected plants, at a high concentration, significantly increased AMF hyphal branches and length. The effect of endophyte exudates on AMF mycelia lenght, varied depending on strain and the concentration used, suggesting a differential interaction between endophyte and AMF species. AMF hyphal branches were increased by fungal exudates in both mycorrhizal species. This study reports the direct and positive effect of Neotyphodium endophytes on AMF pre-infective state, supporting previous information on native grasses. Fil: Novas, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Iannone, Leopoldo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Godeas, Alicia Margarita. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales; Argentina Fil: Scervino, Jose Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales; Argentina

In the present work, we studied the presence of “epichloe/neotyphodium endophytes” in native grasses from Argentina. An extensive area of this country, representative of several different environments, was studied. Nine new host species of the genera Briza, Bromus and Poa were observed to be infected with asexual seed-borne endophytes. Epichloe stromata were not observed on any grass species. The incidence of infection in natural populations was highly variable among host species and among populations of the same host species. Morphological characterization revealed differences among the endophytes of different host species and among endophytes of different populations of the same host species. We also summarize the previous knowledge and present unpublished data on host diversity and distribution of these fungi in Argentina. Our results are discussed and compared with previous studies on endophytes in Argentina. This work supports the hypothesis that sexual species (Epichloe) are not present in the southern hemisphere, and suggests the existence of a high diversity of asexual endophytes in South America.

Background and aims: Plants often establish multiple simultaneous symbiotic associations with different micro-organisms; however, the way in which each symbiont affects the other symbionts and the effects of these multiple interactions on plant performance are not well understood. The aim of this study was to evaluate how two different asexual Epichloë species modulate the establishment of arbuscular mycorrhizal fungi (AMF) in a wild forage grass under different soil fertilization levels. Methods: We performed a completely randomized 12-month-long field experiment to evaluate the effect of two B. auleticus-endophyte ecotypes and two soil fertilization levels on the colonization of AMF, in seedlings and adult plants. Plant biomass and reproductive tillers production were also measured. Results: The symbiosis, measured as the total extent of AM fungal colonization and frequency of arbuscules was significantly higher in Epichloë-infected plants and was not affected by fertilization either in seedlings or in adult plants. Plant biomass was increased by fertilization but no differences were observed due to the endophytic status. However, E+ plants produced more panicles than their E− counterparts. Conclusions: Our findings strongly support the hypothesis of positive association between Epichloë endophytes and AMF in wild grasses, making this model important for agronomic improvement. Fil: Vignale, Maria Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Iannone, Leopoldo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Pinget, Albertina Daniela. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Entre Ríos. Estación Experimental Agropecuaria Concepción del Uruguay; Argentina Fil: De Battista, José P.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Entre Ríos. Estación Experimental Agropecuaria Concepción del Uruguay; Argentina Fil: Novas, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina

Grazing by exotic herbivores on native vegetation in Patagonian steppes has led to the deterioration of forage resources, where grasses are replaced by shrubs and preferred grasses by non-preferred ones. In this region, Hordeum comosum is one of the widely spread native-grasses highly preferred by sheep. Contrary to other preferred grasses, H. comosum establishes symbiosis with vertically-transmitted fungi of genus Epichloë. However, the level of incidence of the fungus and the phylogenetic diversity of the endophyte remained unclear. We found that endophyte incidence ranged from 0 to 100%, with higher incidence in populations from more arid sites. This would suggest an endophyte-conferred drought tolerance to host. Although the isolates presented several morphological differences, phylogenetic analyses of tubB and tefA genes separated them into only two lineages. One of these lineages was Epichloëtembladerae, the most common endophyte in temperate grasses of southern South America. Strikingly, the other lineage was a hybrid between Epichloëtyphina and Epichloëamarillans detected for the first time in this part of the world and opening new questions about the grass-endophyte co-evolution. These results represent a starting-point in the potential use of fungal endophytes in breeding programs and natural grassland restoration in marginal environments. Fil: Iannone, Leopoldo Javier. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Plantas Tóxicas y Medicinales, Metabolismo de Compuestos Sintéticos y Naturales - Hongos que Intervienen en la Degradación Biológica; Argentina. Universidad de Buenos Aires. Facultad de Ingeniería. Departamento de Ingeniería Química; Argentina Fil: Irisarri, Jorge Gonzalo Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; Argentina. Universidad de Buenos Aires. Facultad de Agronomía. Departamento de Produccion Animal. Cátedra de Forrajicultura; Argentina Fil: Mc Cargo, Patricia Débora. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Plantas Tóxicas y Medicinales, Metabolismo de Compuestos Sintéticos y Naturales - Hongos que Intervienen en la Degradación Biológica; Argentina Fil: Perez, Luis Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; Argentina. Universidad de Buenos Aires. Facultad de Agronomia. Departamento de Recursos Naturales y Ambiente. Catedra de Ecologia; Argentina Fil: Gundel, Pedro Emilio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; Argentina. Universidad de Buenos Aires. Facultad de Agronomia. Departamento de Recursos Naturales y Ambiente. Catedra de Ecologia; Argentina

Background: Soil is among the most diverse and complex environments in the world. Soil microorganisms play an essential role in biogeochemical cycles and affect plant growth and crop production. However, our knowledge of the relationship between species-assemblies and soil ecosystem processes is still very limited. The aim of this study was to generate a comprehensive metagenomic survey to evaluate the effect of high-input agricultural practices on soil microbial communities. Results: We collected soil samples from three different areas in the Argentinean Pampean region under three different types of land uses and two soil sources (bulk and rhizospheric). We extracted total DNA from all samples and also synthetized cDNA from rhizospheric samples. Using 454-FLX technology, we generated 112 16S ribosomal DNA and 14 16S ribosomal RNA amplicon libraries totaling 1.3 M reads and 36 shotgun metagenome libraries totaling 17.8 million reads (7.7 GB). Our preliminary results suggested that water availability could be the primary driver that defined microbial assemblages over land use and soil source. However, when water was not a limiting resource (annual precipitation >800 mm) land use was a primary driver. Conclusion: This was the first metagenomic study of soil conducted in Argentina and our datasets are among the few large soil datasets publicly available. The detailed analysis of these data will provide a step forward in our understanding of how soil microbiomes respond to high-input agricultural systems, and they will serve as a useful comparison with other soil metagenomic studies worldwide., Instituto de Biotecnologia y Biologia Molecular

The incidence of epichloid endophytes in populations of wild grasses is usually variable, and the knowledge about distribution patterns and how environmental factors affect such an incidence is limited. Here we performed a broad scale survey data to study whether the distribution patterns and the incidence of vertically-transmitted endophytes in populations of two native grasses from South-America, Poa lanuginosa Poir. and Poa bonariensis (Lam.) Kunth., are associated with environmental characteristics. We also characterized the endophytes from different populations to establish if the genotype of the endophytes is also correlated with environmental variables. The incidence of endophytes ranged from 0 to 100 % in both host species. In P. lanuginosa, endophytes were only found in populations on sandy coastal dunes and their incidence was positively associated with winter regime rainfall and soil water availability in the growing season. In P. bonariensis, endophytes were only found in populations in xerophytic forests and their incidence was highly associated with plant community. The distributions of infested populations suggested that the endophytes are not found in those areas with the most favorable or most stressing growth conditions accordingly to climatic or edaphical characteristics. Only the vertically transmitted hybrid endophyte species Neotyphodium tembladerae was detected in both host species. Under the hypothesis of vertical transmission, these results suggested that the endophyte should have been lost in endophyte free populations but is maintained in populations established in environments presenting moderate stress as salinity or short drought periods. Fil: Iannone, Leopoldo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Mc Cargo, Patricia Débora. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Giussani, Liliana Mónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica Darwinion. Academia Nacional de Ciencias Exactas, Físicas y Naturales. Instituto de Botánica Darwinion; Argentina Fil: Schardl, Christopher L.. University Of Kentucky; Estados Unidos

The fungi of genus Neotyphodium are systemic, constitutive, symbionts of grasses of subfamily Pooideae. In the southern hemisphere most of these asexual endophytes are the result of the hybridization between two sexual species, Epichlo" festucae and E. typhina, from the northern hemisphere. However the ancestral sexual species have not been detected in this region. Several grasses from Argentina are infected by Neotyphodium species. These endophytes are in general very similar macro- and micromorphologically and phylogenetically conform to species N. tembladerae. However the Neotyphodium spp. endophytes of some hosts, Bromus auleticus and Poa spicifomis var. spiciformis, have not been included in this species. In this work we studied the incidence and characterized the diversity of Neotyphodium species in populations of the native grass Bromus auleticus from Argentina. The incidence of endophytes was 100% in all populations investigated. Two groups of endophytes were differentiated by their morphologies, growth rates, conidial ontogenies and by relative resistance to the fungicide benomyl. Phylogenetic trees inferred from tefA and tubB intron sequences indicated that both N. tembladerae and the novel morphotype were hybrids of E. festucae and E. typhina, but the ancestral E. typhina genotype distinguished them. Isolates from plants that inhabit coastal dunes, xerophytic forests, savannahs and hills were similar morphologically and phylogenetically to N. tembladerae, whereas the endophytes from the humid pampa plains conformed to the novel group. We propose the endophyte of Bromus auleticus from humid pampas as a new species, Neotyphodium pampeanum.

The interaction between mycorrhiza and leaf endophytes (Neotyphodium sp.) was studied in three Poa bonariensis populations, a native grass, differing significantly in endophyte infection. The association between endophytes and mycorrhizal fungi colonisation was assessed by analysing plant roots collected from the field. We found that roots from endophyte-infected populations showed a significantly higher frequency of colonisation by mycorrhizal fungi and that soil parameters were not related to endophyte infection or mycorrhiza colonization. In addition, we did not observe significant differences in the number of AM propagules in soils of the three populations sites. We also report the simultaneous development of Paris-type and Arum-type mycorrhiza morphology within the same root systems of P. bonariensis. The co-occurrence of both colonisation types in one and the same root system found in the three populations, which differed in Neotyphodium infection, suggests that foliar endophytes do not determine AM morphology. The percentage of root length colonised by different types of fungal structures (coils, arbuscules, longitudinal hyphae and vesicles) showed significant and positive differences in arbuscular frequency associated with endophyte infection, whereas the much smaller amounts of vesicles and hyphal coils did not differ significantly. Fil: Novas, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Iannone, Leopoldo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Godeas, Alicia Margarita. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina Fil: Cabral, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina