Modeling geographic distribution of arbuscular mycorrhizal fungi from molecular evidence in soils of Argentinean Puna using a maximum entropy approach.

The biogeographic region of Argentinean Puna mainly extends at elevations higher than 3,000 m within the Andean Plateau and hosts diverse ecological communities highly adapted to extreme aridity and low temperatures. Soils of Puna are typically poorly evolved and geomorphology is shaped by drainage networks, resulting in highly vegetated endorheic basins and hypersaline basins known as salar or salt flats. Local communities rely on soil fertility for agricultural practices and on pastures for livestock rearing. From this perspective, investigating the scarcely explored microbiological diversity of these soils as indicators of ecosystems functioning might help to predict the fragility of these harsh environments. In this study we collected soil samples from 28 points, following a nested design within three different macro-habitats, i.e., Puna grassland, hypersaline salar and family-run crop fields. Total fungi and arbuscular mycorrhizal fungi (AMF) occurrence were analyzed using eDNA sequencing. In addition, the significance of soil salinity and organic matter content as significant predictors of AMF occurrence, was assessed through Generalized Linear Mixed Modeling. We also investigated whether intensive grazing by cattle and lama in Puna grasslands may reduce the presence of AMF in these highly disturbed soils, driving or not major ecological changes, but no consistent results were found, suggesting that more specific experiments and further investigations may address the question more specifically. Finally, to predict the suitability for AMF in the different macro-habitats, Species Distribution Modeling (SDM) was performed within an environmental coherent area comprising both the phytogeographic regions of Puna and Altoandino. We modeled AMF distribution with a maximum entropy approach, including bioclimatic and edaphic predictors and obtaining maps of environmental suitability for AMF within the predicted areas. To assess the impact of farming on AMF occurrence, we set a new series of models excluding the cultivated Chaupi Rodeo samples. Overall, SDM predicted a lower suitability for AMF in hypersaline salar areas, while grassland habitats and a wider temperature seasonality range appear to be factors significantly related to AMF enrichment, suggesting a main role of seasonal dynamics in shaping AMF communities. The highest abundance of AMF was observed in Vicia faba crop fields, while potato fields yielded a very low AMF occurrence. The models excluding the cultivated Chaupi Rodeo samples highlighted that if these cultivated areas had theoretically remained unmanaged habitats of Puna and Altoandino, then large-scale soil features and local bioclimatic constraints would likely support a lower suitability for AMF. Using SDM we evidenced the influence of bioclimatic, edaphic and anthropic predictors in shaping AMF occurrence and highlighted the relevance of considering human activities to accurately predict AMF distribution.

Arbuscular Mycorrhizal Fungi from Argentinean Highland Puna Soils Unveiled by Propagule Multiplication.

Low arbuscular-mycorrhizal (AM) sporulation in arid field soils limits our knowledge of indigenous species when diversity studies are based only on spore morphology. Our aim was to use different approaches (i.e., spore morphological approach and PCR-SSCP (single-strand-conformation-polymorphism) analysis after trap plant multiplication strategies to improve the knowledge of the current richness of glomalean AM fungi (Glomerales; Glomeromycota) from the Argentine Puna. Indigenous propagules from two pristine sites at 3870 and 3370 m of elevation were multiplied using different host plants; propagation periods (2-6 months), and subculture cycles (1; 2; or 3) from 5 to 13 months. The propagule multiplication experiment allowed the detection of different glomoid taxa of Funneliformis spp. and Rhizoglomus spp., which were considered cryptic species since they had never been found in Puna soils before. On the other hand; almost all the generalist species previously described were recovered from cultures; except for Glomus ambisporum. Both plant host selection and culture times are critical for Glomerales multiplication. The SSCP analysis complemented the morphological approach and showed a high variability of Glomus at each site; revealing the presence of Funneliformis mosseae. This study demonstrates that AMF trap culture (TC) is a useful strategy for improving the analysis of AM fungal diversity/richness in the Argentinean highlands.

Impact of land use history on the arbuscular mycorrhizal fungal diversity in arid soils of Argentinean farming fields.

Arbuscular mycorrhizal fungi (AMF) are a key soil functional group, with an important potential to increase crop productivity and sustainable agriculture including food security. However, there is clear evidence that land uses, crop rotations and soil features affect the AMF diversity and their community functioning in many agroecosystems. So far, the information related to AMF biodiversity in ecosystems like the Argentinean Puna, an arid high plateau where plants experience high abiotic stresses, is still scarce. In this work, we investigated morphological and molecular AMF diversity in soils of native corn, bean and native potato Andean crops, under a familiar land use, in Chaupi Rodeo (Jujuy, Argentina), without agrochemical supplements but with different histories of crop rotation. Our results showed that AMF morphological diversity was not only high and variable among the three different crop soils but also complemented by Illumina MiSeq data. The multivariate analyses highlighted that total fungal diversity is significantly affected by the preceding crop plants and the rotation histories, more than from the present crop species, while AMF communities are significantly affected by preceding crop only in combination with the effect of nitrogen and calcium soil concentration. This knowledge will give useful information on appropriate familiar farming.

Effect of Arbuscular Mycorrhizal Colonization on Cadmium-Mediated Oxidative Stress in Glycine max (L.) Merr.

Cadmium is a heavy metal (HM) that inhibits plant growth and leads to death, causing great losses in yields, especially in Cd hyperaccumulator crops such as Glycine max (L.) Merr. (soybean), a worldwide economically important legume. Furthermore, Cd incorporation into the food chain is a health hazard. Oxidative stress (OS) is a plant response to abiotic and biotic stresses with an intracellular burst of reactive oxygen species (ROS) that causes damage to lipids, proteins, and DNA. The arbuscular mycorrhizal fungal (AMF) association is a plant strategy to cope with HM and to alleviate OS. Our aim was to evaluate the mitigation effects of mycorrhization with AMF Rhizophagus intraradices on soybean growth, nutrients, Cd accumulation, lipid peroxidation, and the activity of different antioxidant agents under Cd (0.7-1.2 mg kg-1 bioavailable Cd) induced OS. Our results suggest that glutathione may act as a signal molecule in a defense response to Cd-induced OS, and mycorrhization may avoid Cd-induced growth inhibition and reduce Cd accumulation in roots. It is discussed that R. intraradices mycorrhization would act as a signal, promoting the generation of a soybean cross tolerance response to Cd pollution, therefore evidencing the potential of this AMF association for bioremediation and encouragement of crop development, particularly because it is an interaction between a worldwide cultivated Cd hyperaccumulator plant and an AMF-HM-accumulator commonly present in soils.

Arbuscular mycorrhizas and dark septate endophytes associated with grasses from the Argentine Puna.

The Andean Puna is an arid, high-elevation plateau in which plants such as grasses experience high abiotic stress and distinctive environmental conditions. We assessed colonization by arbuscular mycorrhizal fungi (AMF) and dark septate endophytes (DSE) in the roots of 20 native grass species and examined the relationship between root-associated fungi (AMF and DSE) as a function of the elevation of study sites, the photosynthetic pathways of the grass hosts, and the hosts' life cycles. In general, grasses were co-colonized by AMF and DSE and the colonization by AMF and DSE was not extensive. The extension of colonization of AMF and that of DSE were positively correlated, as were number of arbuscules and DSE colonization extension. The extension of AMF colonization differed among sites with different elevations, but DSE colonization was similar across sites. Overall, AMF and DSE patterns shifted as a function of elevation in most grass species, with no general trends observed with respect to host photosynthetic pathway or life cycle. In general, our observations differ from previous studies in the Northern Hemisphere. Variation among sites in AMF and DSE colonization was greater than variation that could be explained by the other factors considered here, suggesting a strong influence of environmental factors. We predict that both AMF and DSE may have established synergistic and beneficial associations with grasses in these distinctive and harsh ecosystems.

Plant functional traits and phylogenetic relatedness explain variation in associations with root fungal endophytes in an extreme arid environment.

Since root endophytes may ameliorate drought stress, understanding which plants associate with endophytes is important, especially in arid ecosystems. Here, the root endophytes were characterized of 42 plants from an arid region of Argentina. Colonization by arbuscular mycorrhizal fungi (AMF) and dark septate endophytes (DSEs) was related to plant functional type (PFT), family, and phylogenetic relatedness. Overall, three main findings were observed. Firstly, only moderate levels of endophyte associations were found across all taxa (e.g., most Poaceae were not colonized by endophytes despite numerous accounts of colonization by AMF and DSEs). We determined 69% of plant taxa associated with some form of root endophyte but levels were lower than other regional studies. Secondly, comparisons by PFT and phylogeny were often qualitatively similar (e.g., succulents and Portulacineae consistently lacked AMF; variation occurred among terrestrial vs. epiphytic bromeliads) and often differed from comparisons based on plant family. Thirdly, comparisons by plant family often failed to account for important variation either within families (e.g., Bromeliaceae and Poaceae) or trait conservatism among related families (i.e., Rosidae consistently lacked DSEs and Portulacineae lacked AMF). This study indicates the value of comparing numerous taxa based on PFTs and phylogenetic similarity. Overall, the results suggest an uncertain benefit of endophytes in extremely arid environments where plant traits like succulence may obviate the need to establish associations.

Colonization of native Andean grasses by arbuscular mycorrhizal fungi in Puna: a matter of altitude, host photosynthetic pathway and host life cycles.

The relationships of altitude, host life cycle (annual or perennial) and photosynthetic pathway (C(3) or C(4) ) with arbuscular mycorrhiza (AM) root colonization were analysed in 35 species of Andean grasses. The study area is located in north-western Argentina along altitudinal sites within the Puna biogeographical region. Twenty-one sites from 3320 to 4314 m were sampled. Thirty-five grasses were collected, and the AM root colonization was quantified. We used multivariate analyses to test emerging patterns in these species by considering the plant traits and variables of AM colonization. Pearson's correlations were carried out to evaluate the specific relationships between some variables. Most grasses were associated with AM, but the colonization percentages were low in both C(3) and C(4) grasses. Nevertheless, the AM root colonization clearly decreased as the altitude increased. This distinctive pattern among different species was also observed between some of the populations of the same species sampled throughout the sites. An inverse relationship between altitude and AM colonization was found in this Southern Hemisphere Andean system. The effect of altitude on AM colonization seems to be more related to the grasses' photosynthetic pathway than to life cycles. This study represents the first report for this biogeographical region.

Arbuscular mycorrhizal fungi in a mountain grassland II: Seasonal variation of colonization studied, along with its relation to grazing and metabolic host type.

The relationships among seasons, host metabolic type, grazing and arbuscular mycorrhizal colonization were analyzed in a high South American native grassland. This study investigated seasonal changes and grazing effects on the symbiotic endomycorrhizal interaction in 5 Poaceae [C(3) metabolic pathway: Briza subaristata Lam., Deyeuxia hieronymi (Hack.) Türpe and Poa stuckertii (Hack.) Parodi; with C(4) metabolic pathway: Eragrostis lugens Nees and Sorghastrum pellitum (Hack.) Parodi; and a Rosaceae (Alchemilla pinnata Ruíz & Pav.)]. All hosts were dominant species in the mountain grassland in central Argentina. It was found that the seasons markedly influenced endomycorrhizal colonization, whereas grazing did not affect this interaction. C(4) grasses presented the highest root colonization. Hosts Briza subaristata (C(3) metabolic pathway) and Sorghastrum pellitum (C(4) metabolic pathway) showed Arum- and Paris-type colonization and intermediate forms.

Native arbuscular mycorrhizal fungi (AMF) from mountain grassland (Cordoba, Argentina) I. Seasonal variation of fungal spore diversity.

Arbuscular mycorrhizal fungi (AMF) were studied in the rhizosphere of 3 Poaceae with metabolic pathway C(3) (Briza subaristata Lam., Deyeuxia hieronymi (Hack.) Türpe and Poa stuckertii (Hack.) Parodi), 2 Poaceae with C(4) metabolic type (Eragrostis lugens Nees and Sorghastrum pellitum (Hack.) Parodi.), and a Rosaceae (Alchemilla pinnata Ruíz & Pav.) from a natural mountain grassland in Central Argentina (South America). Host species, their metabolic type, seasonal changes, and grazing effects over AM fungal diversity were analyzed. Seventeen mycorrhizal fungi taxa were found, widespread in all families of Glomales. Density of endomycorrhizal fungi was found to be strongly influenced with seasons and host metabolic pathway, although biodiversity (H), richness (S) and evenness (E) did not change. In most cases grazing did not affect these variables.