Four new species of Morchella from the Americas.

Morphological and molecular phylogenetic studies of true morels (Morchella) in North America, the Dominican Republic, Venezuela, Ecuador, and Peru led to the discovery of four undescribed species of Morchella. Two new species in the Elata clade, one from the Dominican Republic, initially distinguished by the informal designation Mel-18, and a newly discovered sister species from northern Arizona, are now recognized. Mel-18 is described as a novel phylogenetically distinct species, M. hispaniolensis. Its sister species from Arizona is described as M. kaibabensis, also recovered as an endophyte of Rocky Mountain juniper. Two additional species in the Esculenta clade, M. peruviana discovered in Peru and M. gracilis (previously reported as Mes-14) from the Dominican Republic, Venezuela, and Ecuador, are described as new. We also demonstrate that scanning electron microscopy (SEM) imaging of ascospores using rehydration/dehydration/critical point drying preparation techniques provides for enhanced resolution of spore wall surfaces, thereby increasing the number of morphological traits available to assess differences among otherwise closely related species.

Tropical herbivorous phasmids, but not litter snails, alter decomposition rates by modifying litter bacteria.

Consumers can alter decomposition rates through both feces and selective feeding in many ecosystems, but these combined effects have seldom been examined in tropical ecosystems. Members of the detrital food web (litter-feeders or microbivores) should presumably have greater effects on decomposition than herbivores, members of the green food web. Using litterbag experiments within a field enclosure experiment, we determined the relative effects of common litter snails (Megalomastoma croceum) and herbivorous walking sticks (Lamponius portoricensis) on litter composition, decomposition rates, and microbes in a Puerto Rican rainforest, and whether consumer effects were altered by canopy cover presence. Although canopy presence did not alter consumers' effects, focal organisms had unexpected influences on decomposition. Decomposition was not altered by litter snails, but herbivorous walking sticks reduced leaf decomposition by about 50% through reductions in high quality litter abundance and, consequently, lower bacterial richness and abundance. This relatively unexplored but potentially important link between tropical herbivores, detritus, and litter microbes in this forest demonstrates the need to consider autotrophic influences when examining rainforest ecosystem processes.

Diversity of benthic biofilms along a land use gradient in tropical headwater streams, Puerto Rico.

The properties of freshwater ecosystems can be altered, directly or indirectly, by different land uses (e.g., urbanization and agriculture). Streams heavily influenced by high nutrient concentrations associated with agriculture or urbanization may present conditions that can be intolerable for many aquatic species such as macroinvertebrates and fishes. However, information with respect to how benthic microbial communities may respond to changes in stream ecosystem properties in relation to agricultural or urban land uses is limited, in particular for tropical ecosystems. In this study, diversity of benthic biofilms was evaluated in 16 streams along a gradient of land use at the Turabo watershed in Puerto Rico using terminal restriction fragment length polymorphism. Diversity indices and community structure descriptors (species richness, Shannon diversity, dominance and evenness) were calculated for both bacteria and eukaryotes for each stream. Diversity of both groups, bacteria and eukaryotes, did not show a consistent pattern with land use, since it could be high or low at streams dominated by different land uses. This suggests that diversity of biofilms may be more related to site-specific conditions rather than watershed scale factors. To assess this contention, the relationship between biofilm diversity and reach-scale parameters (i.e., nutrient concentrations, canopy cover, conductivity, and dissolved oxygen) was determined using the Akaike Information Criterion (AIC(c)) for small sample size. Results indicated that nitrate was the variable that best explained variations in biofilm diversity. Since nitrate concentrations tend to increase with urban land use, our results suggest that urbanization may indeed increase microbial diversity indirectly by increasing nutrients in stream water.

Fungal communities of young and mature hypersaline microbial mats.

Microbial mats are a laminated organic-sedimentary ecosystem, found in a wide range of habitats. Fluctuating diel and seasonal physicochemical gradients characterize these ecosystems, resulting in both strata and microenvironments that harbor specific microbial communities. This study was undertaken to compare two types of microbial mats across seasons to further understand the structure of fungal communities in hypersaline microbial mats and their seasonal dynamics. The structure and diversity of fungal communities was documented in young transient and mature hypersaline microbial mats from a tropical region (Puerto Rico) using one culture-dependent and three culture-independent molecular techniques based on the internal transcribed spacer (ITS) region of ribosomal DNA: terminal restriction fragment length polymorphism (TRFLP), denaturing gradient gel electrophoresis (DGGE) and clone libraries. Two microbial mats (one young and transient, one mature) were sampled in Nov 2007 (wet season), Jan 2008 (intermediate season) and Mar 2008 (dry season) in the Cabo Rojo Solar Salterns on the southwestern coast of Puerto Rico. Traditional and molecular techniques revealed strong spatial and temporal heterogeneities in both microbial mats. Higher abundance of isolates and phylotypes were observed during the wet season, and diversity decreased from the top (oxic) to the bottom (anoxic) layers in both seasons. Some of the species isolated belong to the genera Aspergillus, Cladosporium, Hortaea, Pichia and Wallemia, which often are isolated from hypersaline environments. The most abundant clones belong to Acremonium strictum and Cladosporium halotolerans, which were not isolated in pure culture. The differences observed using culture-based and molecular techniques demonstrates the need of combining methods to study the diversity of fungi in a given substrate.

Microbial mats: an ecological niche for fungi.

Fungi were documented in tropical hypersaline microbial mats and their role in the degradation of complex carbohydrates (exopolymeric substance - EPS) was explored. Fungal diversity is higher during the wet season with Acremonium, Aspergillus, Cladosporium, and Penicillium among the more common genera. Diversity is also higher in the oxic layer and in young and transient mats. Enrichments with xanthan (a model EPS) show that without antibiotics (full community) degradation is faster than enrichments with antibacterial (fungal community) and antifungal (bacterial community) agents, suggesting that degradation is performed by a consortium of organisms (bacteria and fungi). The combined evidence from all experiments indicates that bacteria carried out approximately two-third of the xanthan degradation. The pattern of degradation is similar between seasons and layers but degradation is faster in enrichments from the wet season. The research suggests that fungi thrive in these hypersaline consortia and may participate in the carbon cycle through the degradation of complex carbohydrates.

Characterization of fungi from hypersaline environments of solar salterns using morphological and molecular techniques.

The Cabo Rojo Solar Salterns located on the southwest coast of Puerto Rico are composed of two main ecosystems (i.e., salt ponds and microbial mats). Even though these locations are characterized by high solar radiation (mean light intensity of 39 mol photons m-2d-1) they harbour a diverse microscopic life. We used morphological and molecular techniques to identify a series of halotolerant fungi. A total of 183 isolates and 36 species were cultured in this study. From the water from the salt ponds, 86 isolates of 26 species were cultured. The halotolerant fungi isolated from water were: Cladosporium cladosporioides, nine Aspergillus sp., five Penicillium sp. and the black yeast Hortaea werneckii. A distinctive isolate with a blue mycelium was cultured from the salt ponds, representing a new species of Periconia based on morphology and rDNA analysis. Forty-four isolates from eight species were cultured from the sediments around the salt ponds. Most of the sediment isolates formed only sterile mycelium, while several were Chaetomium globosum. A total of 53 isolates from 16 species were isolated from the three layers of the microbial mats, of which Aspergillus niger was the most frequent isolate. Phospholipid fatty acid profiles generated from the different layers of the microbial mats indicated that the uppermost layers of the mats contained fungal biomarker, 18:2w6. This fatty acid decreased with depth, the highest concentration was observed in the green upper layer and it disappeared in the black bottom anoxic layer. This correlates with the isolation of fungi using the serial dilution technique. This is the first study that documents the presence of fungi in microbial mats.

Hygrophoraceae (Agaricales) of the Greater Antilles: Hygrocybe subgenus Pseudohygrocybe sections Coccineae and Neohygrocybe.

A key to 17 species in the genus Hygrocybe, subgenus Pseudohygrocybe, sections Coccineae and Neohygrocybe sensu Boertmann is provided for the Greater Antilles. Five new species and five taxa that are new reports for the region are described. The new species in section Coccineae are H. pseudoadonis, H. viridiphylla, and H. zonata. The new species in section Neohygrocybe are H. albomarginata and H. ovinoides. The new reports are H. caespitosa, H. coccinea, H. cf. miniata, H. papillata, and H. subovina. Three new combinations are proposed: Hygrocybe mycenoides, H. papillata and H. subovina.

New species of Psilocybe in the Caribbean, with an emendation of P. guilartensis.

Five new species of Psilocybe from the Caribbean are described: P. caribaea, P. egonii, P. subpsilocybioides, P. zapotecoantillarum and P. zapotecocaribaea. All except P. zapotecocaribaea, which is known only from Martinique, are native to Puerto Rico. Psilocybe guilartensis, previously described from Puerto Rico, is the most commonly collected species of Psilocybe in Puerto Rico. New information on morphology is provided for P. guilartensis, and an emendation of the species circumscription is presented.