Exploring structural and molecular diversity of Ericaceae hair root mycobionts: a comparison between Northern Bohemia and Argentine Patagonia.

Core Ericaceae produce delicate hair roots with inflated rhizodermal cells that host plethora of fungal symbionts. These poorly known mycobionts include various endophytes, parasites, saprobes, and the ericoid mycorrhizal (ErM) fungi (ErMF) that form the ErM symbiosis crucial for the fitness of their hosts. Using microscopy and high-throughput sequencing, we investigated their structural and molecular diversity in 14 different host × site combinations in Northern Bohemia (Central Europe) and Argentine Patagonia (South America). While we found typical ericoid mycorrhiza in all combinations, we did not detect ectomycorrhiza and arbuscular mycorrhiza. Superficial mantles of various thickness formed by non-clamped hyphae were observed in all combinations except Calluna vulgaris from N. Bohemia. Some samples contained frequent intercellular hyphae while others possessed previously unreported intracellular haustoria-like structures linked with intracellular hyphal coils. The 711 detected fungal OTU were dominated by Ascomycota (563) and Basidiomycota (119), followed by four other phyla. Ascomycetes comprised Helotiales (255), Pleosporales (53), Chaetothyriales (42), and other 19 orders, while basidiomycetes Sebacinales (42), Agaricales (28), Auriculariales (7), and other 14 orders. While many dominant OTU from both hemispheres lacked close relatives in reference databases, many were very similar to identical to unnamed sequences from around the world. On the other hand, several significant ericaceous mycobionts were absent in our dataset, incl. Cairneyella, Gamarada, Kurtia, Lachnum, and Leohumicola. Most of the detected OTU could not be reliably linked to a particular trophic mode, and only two could be reliably assigned to the archetypal ErMF Hyaloscypha hepaticicola. Probable ErMF comprised Hyaloscypha variabilis and Oidiodendron maius, both detected only in N. Bohemia. Possible ErMF comprised sebacinoid fungi and several unnamed members of Hyaloscypha s. str. While H. hepaticicola was dominant only in C. vulgaris, this model ErM host lacked O. maius and sebacinoid mycobionts. Hyaloscypha hepaticicola was absent in two and very rare in six combinations from Patagonia. Nine OTU represented dark septate endophytes from the Phialocephala fortinii s. lat.-Acephala applanata species complex, including the most abundant OTU (the only detected in all combinations). Statistical analyses revealed marked differences between N. Bohemia and Patagonia, but also within Patagonia, due to the unique community detected in a Valdivian temperate rainforest. Our results show that the ericaceous hair roots may host diverse mycobionts with mostly unknown functions and indicate that many novel ErMF lineages await discovery. Transhemispheric differences (thousands of km) in their communities may be evenly matched by local differences (scales of km, m, and less).

Contaminantes microbianos en cervezas artesanales embotelladas de la Patagonia andina argentina / Microbial contaminants in bottled craft beer of Andean Patagonia, Argentina

Resumen La actividad cervecera en la Patagonia andina argentina tiene un rol muy importante en la economía de la región; una de las problemáticas que enfrenta, en términos de calidad, son las contaminaciones microbianas. La presencia de bacterias y levaduras contaminantes en la cerveza produce cambios microbiológicos, físicos y químicos, que impactan en sus atributos sensoriales. No obstante, pocas cervecerías establecen criterios y políticas que garanticen la calidad microbiológica de sus productos. El propósito de este trabajo fue estudiar por primera vez la incidencia de contaminantes microbianos en cervezas artesanales embotelladas producidas en la Patagonia andina argentina, además de identificar los principales microorganismos involucrados y determinar posibles relaciones entre los eventos de contaminación y variables fisicoquímicas de la cerveza. Para ello se analizaron 75 cervezas provenientes de 37 cervecerías de 12 localidades andinas. El 69,3% de las muestras analizadas evidenció crecimiento de microorganismos en los medios de cultivo empleados para la detección de contaminantes cerveceros. La bacteria Levilactobacillus brevis y levaduras del género Saccharomyces fueron los principales contaminantes identificados. Se comprobó que las contaminaciones microbianas impactaron sobre el perfil sensorial de la cerveza y que el cambio de pH fue un indicador de contaminación por bacterias lácticas. De cada 10 fábricas estudiadas, 8 presentaron problemas de contaminación, lo que pone en evidencia la necesidad de diseñar estrategias de prevención y control de contaminaciones en microcervecerías.

Abstract The brewing activity in Andean Patagonia plays a very important role in the region's economy, being microbial contamination one of the main problems in terms of quality. The presence of contaminant bacteria and wild yeasts in beer generate microbiological, physical and chemical changes that impact on its sensory attributes. However, few breweries establish criteria and policies to guarantee the quality of their products in a microbiological sense. The purpose of this work was to study for the first time the incidence of microbial contaminants in bottled craft beers from Andean Patagonia, identify the main microorganisms involved and establish relationships between contamination and the physicochemical variables of beer. We analyzed 75 beers from 37 breweries from 12 different Patagonian cities. Our results showed that 69.3% of the analyzed beer exhibited contaminant microorganism growth. Bacteria Levilactobacillus brevis and wild yeasts of Saccharomyces were the main microorganisms responsible for these contaminations. In addition, we found that microbial contamination had an impact on beer sensory profile and also that pH was correlated with the presence of lactic acid bacteria in beer, being an indicator of contamination for these bacteria. In conclusion, we observed that 8 out of 10 breweries studied showed contamination problems, highlighting the need to design prevention and control strategies in microbreweries.

[Microbial contaminants in bottled craft beer of Andean Patagonia, Argentina]. / Contaminantes microbianos en cervezas artesanales embotelladas de la Patagonia andina argentina.

The brewing activity in Andean Patagonia plays a very important role in the region's economy, being microbial contamination one of the main problems in terms of quality. The presence of contaminant bacteria and wild yeasts in beer generate microbiological, physical and chemical changes that impact on its sensory attributes. However, few breweries establish criteria and policies to guarantee the quality of their products in a microbiological sense. The purpose of this work was to study for the first time the incidence of microbial contaminants in bottled craft beers from Andean Patagonia, identify the main microorganisms involved and establish relationships between contamination and the physicochemical variables of beer. We analyzed 75 beers from 37 breweries from 12 different Patagonian cities. Our results showed that 69.3% of the analyzed beer exhibited contaminant microorganism growth. Bacteria Levilactobacillus brevis and wild yeasts of Saccharomyces were the main microorganisms responsible for these contaminations. In addition, we found that microbial contamination had an impact on beer sensory profile and also that pH was correlated with the presence of lactic acid bacteria in beer, being an indicator of contamination for these bacteria. In conclusion, we observed that 8 out of 10 breweries studied showed contamination problems, highlighting the need to design prevention and control strategies in microbreweries.

First record of Rhizoscyphus ericae in Southern Hemisphere's Ericaceae.

Ericoid mycorrhiza is arguably the least investigated mycorrhizal type, particularly when related to the number of potential hosts and the ecosystems they inhabit. Little is known about the global distribution of ericoid mycorrhizal (ErM) fungi, and this holds true even for the prominent ErM mycobiont Rhizoscyphus ericae. Earlier studies suggested R. ericae might be low in abundance or absent in the roots of Southern Hemisphere's Ericaceae, and our previous investigations in two Argentine Patagonian forests supported this view. Here, we revisited the formerly investigated area, albeit at a higher altitude, and screened fungi inhabiting hair roots of Gaultheria caespitosa and Gaultheria pumila at a treeless alpine site using the same methods as previously. We obtained 234 isolates, most of them belonging to Ascomycota. In contrast to previous findings, however, among 37 detected operational taxonomic units (OTUs), OTU 1 (=R. ericae s. str.) comprised the highest number of isolates (87, ∼37 %). Most of the OTUs and isolates belonged to the Helotiales, and 82.5 % of isolates belonged to OTUs shared between both Gaultheria species. At the alpine site, ericoid mycorrhizal fungi dominated, followed by dark septate endophytes and aquatic hyphomycetes probably acting as root endophytes. Our results suggest that the distribution of R. ericae is influenced, among others, by factors related to altitude such as soil type and presence/absence and type of the neighboring vegetation. Our study is the first report on R. ericae colonizing Ericaceae roots in the Southern Hemisphere and extends the known range of this prominent ErM species to NW Patagonia.

Is the prominent ericoid mycorrhizal fungus Rhizoscyphus ericae absent in the Southern Hemisphere's Ericaceae? A case study on the diversity of root mycobionts in Gaultheria spp. from northwest Patagonia, Argentina.

Ericaceae diversity hotspots are in the mountains of the Neotropics and Papua New Guinea, South Africa's fynbos and Southeast Asia but majority of references to their root mycobionts come from the Northern Hemisphere. Here, typical cultivable ericoid mycorrhizal (ErM) fungi comprise Rhizoscyphus ericae, Meliniomyces variabilis, and Oidiodendron maius. It is however unclear whether this is true also for the Southern Hemisphere. Our study focused on cultivable mycobionts from hair roots of Gaultheria mucronata and Gaultheria poeppigii (Ericaceae) from two natural forests in NW Patagonia, Argentina, differing in mycorrhizal preferences of their tree dominants. We detected 62 well-defined OTUs mostly belonging to Helotiales and Hypocreales; the most frequent were Phialocephala fortinii s. l., Pochonia suchlasporia, and Ilyonectria radicicola. Only one out of 257 isolates showed ITS nrDNA similarity to members of the R. ericae aggregate (REA) but was not conspecific with R. ericae, and only five isolates were conspecific with O. maius. Microscopic observations showed that the screened roots were frequently colonized in a manner differing from the pattern typically produced by R. ericae and O. maius. A re-synthesis experiment with selected isolates showed that only O. maius formed colonization resembling ericoid mycorrhiza. Amplification of root fungal DNA with REA-specific and Sebacinaceae-specific primers showed that REA mycobionts were present in some of the screened samples while Sebacinaceae were present in all samples. These results suggest that Gaultheria spp. from NW Patagonia form ericoid mycorrhizae predominantly with the difficult-to-cultivate Sebacinaceae while the incidence of REA is relatively low and may be masked by other most likely non-mycorrhizal cultivable mycobionts.