Unearthing the soil-borne microbiome of land plants.

Plant-soil biodiversity interactions are fundamental for the functioning of terrestrial ecosystems. Yet, the existence of a set of globally distributed topsoil microbial and small invertebrate organisms consistently associated with land plants (i.e., their consistent soil-borne microbiome), together with the environmental preferences and functional capabilities of these organisms, remains unknown. We conducted a standardized field survey under 150 species of land plants, including 58 species of bryophytes and 92 of vascular plants, across 124 locations from all continents. We found that, despite the immense biodiversity of soil organisms, the land plants evaluated only shared a small fraction (less than 1%) of all microbial and invertebrate taxa that were present across contrasting climatic and soil conditions and vegetation types. These consistent taxa were dominated by generalist decomposers and phagotrophs and their presence was positively correlated with the abundance of functional genes linked to mineralization. Finally, we showed that crossing environmental thresholds in aridity (aridity index of 0.65, i.e., the transition from mesic to dry ecosystems), soil pH (5.5; i.e., the transition from acidic to strongly acidic soils), and carbon (less than 2%, the lower limit of fertile soils) can result in drastic disruptions in the associations between land plants and soil organisms, with potential implications for the delivery of soil ecosystem processes under ongoing global environmental change.

Identifying Strategies for Effective Biodiversity Preservation and Species Status of Chilean Amphibians.

Resources are limited in global biodiversity conservation efforts, which emphasizes the significance of setting conservation priorities. Using standardized criteria, we evaluated 58 amphibian species in Chile to determine their conservation priority (CP). Species with insufficient historical data had their values marked as missing. With a median value of p = 1.67, the results demonstrated CP values ranging from p = 0.48 to p = 3.0, classifying species into priority and non-priority groups. Four levels were established for the priority categories: no priority, low priority, medium priority, and high priority. Additionally, the Telmatobiidae and Alsodidae families were identified as two more priority families. Notably, the species with the highest priority were found to be T. halli, T. fronteriensis, T. philippii, T. chusmisensis, A. pehuenche, and Alsodes tumultuosus, where T. philippii and T. fronteriensis have equal priority for conservation at the national level according to the conservation priority analysis. Eight priority families-the Alsodidae, Batrachylidae, Bufonidae, Ceratophryidae, Leptodactylidae, Rhinodermatidae, and Telmatobiidae-were determined, and 14 species-or 24% of the species examined-need further study. Based on the conservation priority analysis, the species T. fronteriensis and T. philippii share the highest priority for conservation at the national level (p = 2.50). With 70% of the amphibians under study being threatened mainly by habitat loss, pollution, and emerging diseases, the creation of conservation categories made the threat assessment process easier. Due to a lack of information on geographic distribution and abundance, quantitatively classifying amphibians in Chile remains difficult. The analysis of conservation priorities and potential extinction threats informs appropriate management strategies.

Genetic control underlying the flowering-drought tolerance trade-off in the Antarctic plant Colobanthus quitensis.

Plants inhabiting environments with stressful conditions often exhibit a low number of flowers, which can be attributed to the energetic cost associated with reproduction. One of the most stressful environments for plants is the Antarctic continent, characterized by limited soil water availability and low temperatures. Induction of dehydrins like those from the COR gene family and auxin transcriptional response repressor genes (IAAs), which are involved in floral repression, has been described in response to water stress. Here, we investigated the relationship between the water deficit-induced stress response and the number of flowers in Colobanthus quitensis plants collected from populations along a latitudinal gradient. The expression levels of COR47 and IAA12 genes in response to water deficit were found to be associated with the number of flowers. The relationship was observed both in the field and growth chambers. Watering the plants in the growth chambers alleviated the stress and stimualted flowering, thereby eliminating the trade-off observed in the field. Our study provides a mechanistic understanding of the ecological constraints on plant reproduction along a water availability gradient. However, further experiments are needed to elucidate the primary role of water availability in regulating resource allocation to reproduction in plants inhibiting extreme environments.

Genetic Diversity, Morphometric Characterization, and Conservation Reassessment of the Critically Endangered Freshwater Snail, Heleobia atacamensis, in the Atacama Saltpan, Northern Chile.

Evaporitic ecosystems of the Atacama Desert contain a rich endemic fauna, including mollusk species. A recent study performed in the freshwater snail Heleobia atacamensis, endemic to the Atacama Saltpan, revealed a strong interdependence of genetic patterns with climatic fluctuations and landscape physiography. The species is currently listed as Critically Endangered at regional scale and as Data Deficient on the International Union for Conservation of Nature (IUCN) Red List. Here, we studied genetic diversity and demographic history of several populations of the species occurring on a connectivity gradient, including snails from new peripherical localities (Peine and Tilomonte), which were compared with topotype specimens. In addition, we reassessed the conservation status using the IUCN Red List categories and criteria considering species-specific idiosyncrasy. Phylogenetic and phylogeographical analyses indicated that snails from Peine and Tilomonte belong to H. atacamensis. We discovered significant differentiation in shell morphology, which was generally greater in geographically isolated populations. We also inferred six genetic clusters and a demographic expansion congruent with the wet periods that occurred at the end of the Pleistocene. Considering the highest risk category obtained, H. atacamensis was reassessed as Endangered at regional scale. Future conservation plans should consider the genetic assemblages as conservation units.

Electromagnetic fields disrupt the pollination service by honeybees.

We assessed the effect that electromagnetic field (EMF) exerts on honeybees' pollination efficiency using field and laboratory experiments. First, we measured levels of gene and protein expression in metabolic pathways involved in stress and behavioral responses elicited by EMF. Second, we assessed the effect of EMF on honeybee behavior and seed production by the honeybee-pollinated California poppy and, lastly, by measuring the consequences of pollination failure on plants' community richness and abundance. EMF exposure exerted strong physiological stress on honeybees as shown by the enhanced expression of heat-shock proteins and genes involved in antioxidant activity and affected the expression levels of behavior-related genes. Moreover, California poppy individuals growing near EMF received fewer honeybee visits and produced fewer seeds than plants growing far from EMF. Last, we found a hump-shaped relationship between EMF and plant species richness and plant abundance. Our study provides conclusive evidence of detrimental impacts of EMF on honeybee's pollination behavior, leading to negative effects on plant community.

Soil contamination in nearby natural areas mirrors that in urban greenspaces worldwide.

Soil contamination is one of the main threats to ecosystem health and sustainability. Yet little is known about the extent to which soil contaminants differ between urban greenspaces and natural ecosystems. Here we show that urban greenspaces and adjacent natural areas (i.e., natural/semi-natural ecosystems) shared similar levels of multiple soil contaminants (metal(loid)s, pesticides, microplastics, and antibiotic resistance genes) across the globe. We reveal that human influence explained many forms of soil contamination worldwide. Socio-economic factors were integral to explaining the occurrence of soil contaminants worldwide. We further show that increased levels of multiple soil contaminants were linked with changes in microbial traits including genes associated with environmental stress resistance, nutrient cycling, and pathogenesis. Taken together, our work demonstrates that human-driven soil contamination in nearby natural areas mirrors that in urban greenspaces globally, and highlights that soil contaminants have the potential to cause dire consequences for ecosystem sustainability and human wellbeing.

Distribution, conservation status and proposed measures for preservation of Radiodiscus microgastropods in Chile.

The genus Radiodiscus includes minute terrestrial snails occurring throughout the American continent. We assessed the conservation status of eight poorly known Chilean Radiodiscus species using the International Union for Conservation of Nature (IUCN) and NatureServe categories and criteria. Under the IUCN guidelines the species were assessed using the Criterion B of geographic range, which considers the extent of occurrence (EOO) and area of occupancy (AOO) as subcriteria. For NatureServe we used these two parameters plus the number of occurrences, ecological viability, and threats. Considering species rarity and possible sampling bias, we also used ecological niche modeling to determine climate and environmental tolerances and predict potential species distributions analyzing bioclimatic and geographical layers. Radiodiscus australis, R. coarctatus and R. quillajicola were listed as Critically Endangered by IUCN and NatureServe standards; R. coppingeri, R. flammulatus, R. magellanicus and R. villarricensis as Endangered by both methods; while R. riochicoensis as Endangered by IUCN standards and Vulnerable by NatureServe standards. Niche modeling results indicated that Radiodiscus species respond to different environmental conditions and that the predicted distribution areas contain suitable habitats beyond the current ranges, which may be helpful for future management plans. Nature-based sport tourism, forestry activities, urbanization, roads, pollution, mining, forest fires, livestock, volcanism, tsunamis, soil erosion and introduced species are among the major threats affecting these snails. Based on the low number of occurrences and the threats identified, the most at-risk species are R. coarctatus and R. quillajicola (one record), R. australis (two records) and R. villarricensis (three records); the latter two lacking occurrences within protected areas. Compiling our findings, we propose a list of actions to preserve Chilean Radiodiscus species.

The global distribution and environmental drivers of the soil antibiotic resistome.

BACKGROUND: Little is known about the global distribution and environmental drivers of key microbial functional traits such as antibiotic resistance genes (ARGs). Soils are one of Earth's largest reservoirs of ARGs, which are integral for soil microbial competition, and have potential implications for plant and human health. Yet, their diversity and global patterns remain poorly described. Here, we analyzed 285 ARGs in soils from 1012 sites across all continents and created the first global atlas with the distributions of topsoil ARGs. RESULTS: We show that ARGs peaked in high latitude cold and boreal forests. Climatic seasonality and mobile genetic elements, associated with the transmission of antibiotic resistance, were also key drivers of their global distribution. Dominant ARGs were mainly related to multidrug resistance genes and efflux pump machineries. We further pinpointed the global hotspots of the diversity and proportions of soil ARGs. CONCLUSIONS: Together, our work provides the foundation for a better understanding of the ecology and global distribution of the environmental soil antibiotic resistome. Video Abstract.

Global hotspots for soil nature conservation.

Soils are the foundation of all terrestrial ecosystems1. However, unlike for plants and animals, a global assessment of hotspots for soil nature conservation is still lacking2. This hampers our ability to establish nature conservation priorities for the multiple dimensions that support the soil system: from soil biodiversity to ecosystem services. Here, to identify global hotspots for soil nature conservation, we performed a global field survey that includes observations of biodiversity (archaea, bacteria, fungi, protists and invertebrates) and functions (critical for six ecosystem services) in 615 composite samples of topsoil from a standardized survey in all continents. We found that each of the different ecological dimensions of soils-that is, species richness (alpha diversity, measured as amplicon sequence variants), community dissimilarity and ecosystem services-peaked in contrasting regions of the planet, and were associated with different environmental factors. Temperate ecosystems showed the highest species richness, whereas community dissimilarity peaked in the tropics, and colder high-latitudinal ecosystems were identified as hotspots of ecosystem services. These findings highlight the complexities that are involved in simultaneously protecting multiple ecological dimensions of soil. We further show that most of these hotspots are not adequately covered by protected areas (more than 70%), and are vulnerable in the context of several scenarios of global change. Our global estimation of priorities for soil nature conservation highlights the importance of accounting for the multidimensionality of soil biodiversity and ecosystem services to conserve soils for future generations.

Phylogeography and molecular species delimitation reveal cryptic diversity in Potamolithus (Caenogastropoda: Tateidae) of the southwest basin of the Andes.

The species of the genus Potamolithus inhabiting the southwestern basin of the Andes are difficult to distinguish due to small size and similar shell morphology. Only Potamolithus australis and Potamolithus santiagensis have been traditionally recognized in this region, but the occurrence of several morphologically similar undescribed populations could increase the regional richness. Here we delimit described and potentially undescribed cryptic species of the genus using partial sequences of the mitochondrial cytochrome c oxidase subunit I (COI) gene. Network analysis and diversity indices inferred six highly differentiated haplogroups, many of them sympatric and widespread in the study area. Phylogeographic analyses suggest a scenario of recent diversification and the occurrence of multiple refuges during the successive Pleistocene glaciations. Phylogenetic analysis also recovered six major clades that showed no relationship with physiography. Species delimitation analyses consistently recognized three or four candidate species apart from P. australis and P. santiagensis. Divergence times indicate that speciation of Chilean Potamolithus began at the end of the Pliocene, probably driven by climatic rather than geographic events. Considering the high inter- and intra-basin genetic diversity, conservation efforts should be focused on protecting sympatric taxa in the basins with the highest species richness.

Species and genera in Aleurodiscus sensu lato as viewed from the Southern Hemisphere.

Phylogenetic relationships of 12 species in Aleurodiscus sensu lato (Stereaceae, Russulales) described from the Patagonian forests of Chile and Argentina were investigated based on sequences of nuc rDNA internal transcribed spacer region ITS1-5.8S-ITS2 (ITS) and the D1-D2 domains of nuc 28S rDNA (28S). A new genus and a new species are presented, and 10 new combinations proposed. The genus Gloeosoma is shown to be phylogenetically well supported and morphologically circumscribed; it includes G. vitellinum (type species), G. mirabile, comb. nov., G. zealandicum, comb. nov., and Gloeosoma decorticans, sp. nov., which is newly described from Chile. The new genus Stereodiscus is proposed to accommodate a group of taxa characterized by an austral distribution and morphologically by smooth, thin-walled, amyloid basidiospores and a lack of gloeocystidia and acanthocystidia; three species develop Stereum-like basidiomata and two species present discoid ones. The new genus includes the species formerly known as Aleurodiscus antarcticus, A. limonisporus, A. parmuliformis, A. patagonicus, and A. triviale. Specimens of Stereodiscus parmuliformis (A. parmuliformis) from New Zealand (where it was originally described) and southern Chile are shown to be phylogenetically conspecific, which confirms its presence in Patagonia. Gloeosoma and Stereodiscus are shown to be distantly related to Aleurodiscus s. str. and other genera in Stereaceae. The new combinations Aleurocystidiellum bernicchiae, Aleurocystidiellum hallenbergii, and Acanthobasidium quilae are proposed based on morphology and phylogenetic analyses, and Aleurodiscus cerussatus is shown to be a cryptic species complex.

Symbiotic Interaction Enhances the Recovery of Endangered Tree Species in the Fragmented Maulino Forest.

Beneficial plant-associated microorganisms, such as fungal endophytes, are key partners that normally improve plant survival under different environmental stresses. It has been shown that microorganisms from extreme environments, like those associated with the roots of Antarctica plants, can be good partners to increase the performance of crop plants and to restore endangered native plants. Nothofagus alessandrii and N. glauca, are among the most endangered species of Chile, restricted to a narrow and/or limited distributional range associated mainly to the Maulino forest in Chile. Here we evaluated the effect of the inoculation with a fungal consortium of root endophytes isolated from the Antarctic host plant Colobanthus quitensis on the ecophysiological performance [photosynthesis, water use efficiency (WUE), and growth] of both endangered tree species. We also, tested how Antarctic root-fungal endophytes could affect the potential distribution of N. alessandrii through niche modeling. Additionally, we conducted a transplant experiment recording plant survival on 2 years in order to validate the model. Lastly, to evaluate if inoculation with Antarctic endophytes has negative impacts on native soil microorganisms, we compared the biodiversity of fungi and bacterial in the rhizospheric soil of transplanted individuals of N. alessandrii inoculated and non-inoculated with fungal endophytes. We found that inoculation with root-endophytes produced significant increases in N. alessandrii and N. glauca photosynthetic rates, water use efficiencies and cumulative growth. In N. alessandrii, seedling survival was significantly greater on inoculated plants compared with non-inoculated individuals. For this species, a spatial distribution modeling revealed that, inoculation with root-fungal endophytes could potentially increase the current distributional range by almost threefold. Inoculation with root-fungal endophytes, did not reduce native rhizospheric microbiome diversity. Our results suggest that the studied consortium of Antarctic root-fungal endophytes improve the ecophysiological performance as well as the survival of inoculated trees and can be used as a biotechnological tool for the restoration of endangered tree species.

Isolation and characterization of microsatellites for the endangered endemic tree Nothofagus alessandrii (Nothofagaceae).

Nothofagus alessandrii (Nothofagaceae) is one of the most endangered trees from Chile due to high rates of habitat disturbance caused by human activities. Despite its conservation status, few molecular markers are available to study its population genetic, connectivity and to assist reproduction programs. Thus, the species needs urgent actions to restore its original distribution. Novel polymorphic microsatellites from the genome of N. alessandrii were isolated and characterized using high-through sequencing. A total of 30 primer pairs were synthesized and 18 microsatellites were amplified correctly. Polymorphism and genetic diversity was evaluated in 58 individuals from three populations of N. alessandrii. Sixteen of them were polymorphic and the number of alleles in the pooled sample ranged from 2 to 14, the mean number of alleles was 4.81. The mean values of observed heterozigosity (HO) and excepted heterozygosity (HE) are similar in all studied populations. Linkage disequilibrium was found between a few pairs of loci (five out of 263 tests) suggesting that most of the markers can be considered as independent. Significant deviations from Hardy-Weinberg equilibrium (P < 0.05) were found in four loci probably due to low sampling size. Transferability to the congeneric N. pumilio was successful in only four out of the sixteen polymorphic markers. The microsatellite markers developed in this study will be useful to study the genetic diversity and structure and to develop integrated management plans for the conservation of this endangered species.

Fungal Symbionts Enhance N-Uptake for Antarctic Plants Even in Non-N Limited Soils.

Plant-fungi interactions have been identified as fundamental drivers of the plant host performance, particularly in cold environments where organic matter degradation rates are slow, precisely for the capacity of the fungal symbiont to enhance the availability of labile nitrogen (N) in the plant rhizosphere. Nevertheless, these positive effects appear to be modulated by the composition and amount of the N pool in the soil, being greater when plant hosts are growing where N is scarce as is the case of Antarctic soils. Nevertheless, in some coastal areas of this continent, seabirds and marine mammal colonies exert, through their accumulated feces and urine a strong influence on the edaphic N content surrounding their aggregation points. To evaluate if the fungal symbionts (root endophytes), associated to the only two Antarctic vascular plants Colobanthus quitensis and Deschampsia antarctica, act as N-uptake enhancers, even in such N-rich conditions as those found around animal influence, we assessed, under controlled conditions, the process of N mineralization in soil by the accumulation of NH4 + in the rizhosphere and the biomass accumulation of plants with (E+) and without (E-) fungal symbionts. Complementarily, taking advantage of the isotopic N-fractionation that root-fungal symbionts exert on organic N molecules during its acquisition process, we also determined if endophytes actively participate in the Antarctic plants N-uptake, when inorganic N is not a limiting factor, by estimating the δ15N isotopic signatures in leaves. Overall, symbiotic interaction increased the availability of NH4 + in the rhizosphere of both species. As expected, the enhanced availability of inorganic N resulted in a higher final biomass in E + compared with E- plants of both species. In addition, we found that the positive role of fungal symbionts was also actively linked to the process of N-uptake in both species, evidenced by the contrasting δ15N signatures present in E+ (-0.4 to -2.3‰) relative to E- plants (2.7-3.1‰). In conclusion, despite being grown under rich N soils, the two Antarctic vascular plants showed that the presence of root-fungal endophytes, furthermore enhanced the availability of inorganic N sources in the rhizosphere, has a positive impact in their biomass, remarking the active participation of these endophytes in the N-uptake process for plants inhabiting the Antarctic continent.

Morphological and molecular identification of the invasive freshwater snail Physa acuta (Gastropoda: Physidae) into Llanquihue Lake, Chilean Patagonia.

The sewage snail Physa acuta, native to North America, is an effective invasive species around the world. In Chile, it was first reported in 2014 in the north central area of the country. So far, the species has not been recorded in southern Chile. Sampling performed in 2015 in three localities from Llanquihue Lake, Chilean Patagonia, only provided native freshwater snails. However, new collections performed in February 2018 in the same three sites were successful for physid specimens suggesting a biological invasion entailing a large southward range expansion of these snails. Here we performed morphological, microstructural and phylogenetic analyses to investigate whether the new samples belong to Physa acuta. The shell morphology, male copulatory complex and radula microstructure of the new material agree with those of the sewage snail. The molecular phylogenetic analyses using the cytochrome c oxidase subunit 1 (COI) gene confirmed morphological identification. We suggest to take prompt measures to prevent the expansion of Physa acuta in Llanquihue Lake or nearby aquatic ecosystems.

Root endophytic Penicillium promotes growth of Antarctic vascular plants by enhancing nitrogen mineralization.

Fungal endophyte associations have been suggested as a possible strategy of Antarctic vascular plants for surviving the extreme environmental conditions of Antarctica. However, the mechanisms by which this occurs are still poorly understood. The role of root fungal endophytes in nitrogen mineralization and nutrient uptake, as well as their impact on the performance of Antarctic plants, were studied. We tested root endophytes, isolated from Colobanthus quitensis and Deschampsia antarctica, for lignocellulolytic enzyme production, nitrogen mineralization, and growth enhancement of their host plants. Penicillium chrysogenum and Penicillium brevicompactum were identified using a molecular approach as the main root endophytes inhabiting C. quitensis and D. antarctica, respectively. Both root endophytes were characterized as psychrophilic fungi displaying amylase, esterase, protease, cellulase, hemicellulase, phosphatase and urease enzymatic activities, mainly at 4 °C. Moreover, the rates and percentages of nitrogen mineralization, as well as the final total biomass, were significantly higher in symbiotic C. quitensis and D. antarctica individuals. Our findings suggest that root endophytes exert a pivotal ecological role based not only to breakdown different nutrient sources but also on accelerating nitrogen mineralization, improving nutrient acquisition, and therefore promoting plant growth in Antarctic terrestrial ecosystems.

Antarctic root endophytes improve physiological performance and yield in crops under salt stress by enhanced energy production and Na+ sequestration.

Climatic change is pointed as one of the major challenges for global food security. Based on current models of climate change, reduction in precipitations and in turn, increase in the soil salinity will be a sharp constraint for crops productivity worldwide. In this context, root fungi appear as a new strategy to improve plant ecophysiological performance and crop yield under abiotic stress. In this study, we evaluated the impact of the two fungal endophytes Penicillium brevicompactum and P. chrysogenum isolated from Antarctic plants on nutrients and Na+ contents, net photosynthesis, water use efficiency, yield and survival in tomato and lettuce, facing salinity stress conditions. Inoculation of plant roots with fungal endophytes resulted in greater fresh and dry biomass production, and an enhanced survival rate under salt conditions. Inoculation of plants with the fungal endophytes was related with a higher up/down-regulation of ion homeostasis by enhanced expression of the NHX1 gene. The two endophytes diminished the effects of salt stress in tomato and lettuce, provoked a higher efficiency in photosynthetic energy production and an improved sequestration of Na+ in vacuoles is suggested by the upregulating of the expression of vacuolar NHX1 Na+/H+ antiporters. Promoting plant-beneficial interactions with root symbionts appears to be an environmentally friendly strategy to mitigate the impact of climate change variables on crop production.

Functional roles of microbial symbionts in plant cold tolerance.

In this review, we examine the functional roles of microbial symbionts in plant tolerance to cold and freezing stresses. The impacts of symbionts on antioxidant activity, hormonal signaling and host osmotic balance are described, including the effects of the bacterial endosymbionts Burkholderia, Pseudomonas and Azospirillum on photosynthesis and the accumulation of carbohydrates such as trehalose and raffinose that improve cell osmotic regulation and plasma membrane integrity. The influence of root fungal endophytes and arbuscular mycorrhizal fungi on plant physiology at low temperatures, for example their effects on nutrient acquisition and the accumulation of indole-3-acetic acid and antioxidants in tissues, are also reviewed. Meta-analyses are presented showing that aspects of plant performance (shoot biomass, relative water content, sugar and proline concentrations and Fv /Fm ) are enhanced in symbiotic plants at low (-1 to 15 °C), but not at high (20-26 °C), temperatures. We discuss the implications of microbial symbionts for plant performance at low and sub-zero temperatures in the natural environment and propose future directions for research into the effects of symbionts on the cold and freezing tolerances of plants, concluding that further studies should routinely incorporate symbiotic microbes in their experimental designs.