Carbon and phosphorus exchange rates in arbuscular mycorrhizas depend on environmental context and differ among co-occurring plants.

Phosphorus (P) for carbon (C) exchange is the pivotal function of arbuscular mycorrhiza (AM), but how this exchange varies with soil P availability and among co-occurring plants in complex communities is still largely unknown. We collected intact plant communities in two regions differing c. 10-fold in labile inorganic P. After a 2-month glasshouse incubation, we measured 32P transfer from AM fungi (AMF) to shoots and 13C transfer from shoots to AMF using an AMF-specific fatty acid. AMF communities were assessed using molecular methods. AMF delivered a larger proportion of total shoot P in communities from high-P soils despite similar 13C allocation to AMF in roots and soil. Within communities, 13C concentration in AMF was consistently higher in grass than in blanketflower (Gaillardia aristata Pursh) roots, that is P appeared more costly for grasses. This coincided with differences in AMF taxa composition and a trend of more vesicles (storage structures) but fewer arbuscules (exchange structures) in grass roots. Additionally, 32P-for-13C exchange ratios increased with soil P for blanketflower but not grasses. Contrary to predictions, AMF transferred proportionally more P to plants in communities from high-P soils. However, the 32P-for-13C exchange differed among co-occurring plants, suggesting differential regulation of the AM symbiosis.

Variation Among Japanese Miso Breweries in Indoor Microbiomes is Mainly Ascribed to Variation in Type of Indoor Surface.

Miso is a microbially-fermented soybean food. The miso brewery indoor microbiome contributes to miso fermentation. Japanese breweries are not climate-controlled, so indoor spaces are strongly affected by the prevailing climate. Because climate influences microorganism distribution, our first hypothesis is that latitude, as a proxy for climate, is a major determinant of brewery indoor microbiome structure. Breweries vary in interior surface materials and in the way operations (steaming, processing, fermenting) are apportioned among rooms. Therefore, our second hypothesis is that more variability in indoor microbiomes exists among breweries than can be ascribed to a latitudinal gradient. Most miso produced today is inoculated with commercial microbial strains to standardize fermentation. If commercial strains outcompete indigenous microbes for membership in the indoor microbiome, this practice may homogenize indoor microbiomes among regions or breweries. Therefore, our third hypothesis is that inoculant fungal species dominate indoor fungal communities and make it impossible to distinguish communities among breweries or across their latitudinal gradient. We tested these hypotheses by sampling indoor surfaces in several breweries across a latitudinal gradient in Japan. We found that latitude had a significant but relatively small impact on indoor fungal and bacterial communities, that the effect of brewery was large relative to latitude, and that inoculant fungi made such small contributions to the indoor microbiome that distinctions among breweries and along the latitudinal gradient remained apparent. Recently, the Japanese Ministry of Agriculture, Forestry and Fisheries specified fungal inoculants to standardize miso production. However, this may not be possible so long as the indoor microbiome remains uncontrolled.

Coupled anaerobic methane oxidation and metal reduction in soil under elevated CO2.

Continued current emissions of carbon dioxide (CO2 ) and methane (CH4 ) by human activities will increase global atmospheric CO2 and CH4 concentrations and surface temperature significantly. Fields of paddy rice, the most important form of anthropogenic wetlands, account for about 9% of anthropogenic sources of CH4 . Elevated atmospheric CO2 may enhance CH4 production in rice paddies, potentially reinforcing the increase in atmospheric CH4 . However, what is not known is whether and how elevated CO2 influences CH4 consumption under anoxic soil conditions in rice paddies, as the net emission of CH4 is a balance of methanogenesis and methanotrophy. In this study, we used a long-term free-air CO2 enrichment experiment to examine the impact of elevated CO2 on the transformation of CH4 in a paddy rice agroecosystem. We demonstrate that elevated CO2 substantially increased anaerobic oxidation of methane (AOM) coupled to manganese and/or iron oxides reduction in the calcareous paddy soil. We further show that elevated CO2 may stimulate the growth and metabolism of Candidatus Methanoperedens nitroreducens, which is actively involved in catalyzing AOM when coupled to metal reduction, mainly through enhancing the availability of soil CH4 . These findings suggest that a thorough evaluation of climate-carbon cycle feedbacks may need to consider the coupling of methane and metal cycles in natural and agricultural wetlands under future climate change scenarios.

On Holobionts, Holospecies, and Holoniches: the Role of Microbial Symbioses in Ecology and Evolution.

My goal in writing this is to increase awareness of the roles played by microbial symbionts in eukaryote ecology and evolution. Most eukaryotes host one or more species of symbiotic microorganisms, including prokaryotes and fungi. Many of these have profound impacts on the biology of their hosts. For example, microbial symbionts may expand the niches of their hosts, cause rapid adaptation of the host to the environment and re-adaptation to novel conditions via symbiont swapping, facilitate speciation, and fundamentally alter our concept of the species. In some cases, microbial symbionts and multicellular eukaryote hosts have a mutual dependency, which has obvious conservation implications. Hopefully, this contribution will stimulate a reevaluation of important ecological and evolutionary concepts including niche, adaptation, the species, speciation, and conservation of multicellular eukaryotes.

Many foliar endophytic fungi of Quercus gambelii are capable of psychrotolerant saprotrophic growth.

Many endophytic fungi have the potential to function as saprotrophs when living host tissues senesce and enter the litter pool. The consumption of plant litter by fungi obviously requires moisture but, in the arid, western USA, the native range of Quercus gambelii Nutt., most of the precipitation occurs during the coldest months of the year. Therefore, we hypothesized that the endophytic fungi of Q. gambelii have the potential to function as psychrotolerant saprotrophs, which we defined in this study as an organism capable of significant growth on leaf litter at 5°C. We further hypothesized that a tradeoff exists between growth of endophytic fungi at 5°C and at 17°C such that fungal isolates are either cold- or warm-temperature specialists. Consistent with our first hypothesis, we found that 36 of our 40 isolates consumed leaf litter at 5°C, but there was a surprisingly high degree of variability among isolates in this ability, even among isolates of a given species. Contrary to our second hypothesis, there was no tradeoff between saprotrophic growth at 5°C and saprotrophic growth at 17°C. Indeed, the isolates that grew poorly as saprotrophs at 5°C were generally those that grew poorly as saprotrophs at 17°C. By virtue of being endophytic, endophytic fungi have priority in litter over decomposer fungi that colonize plant tissues only after they enter the litter pool. Moreover, by virtue of being psychrotolerant, some endophytic fungi may function as saprotrophs during the cold months of the year when moisture is temporarily available. Therefore, we suggest that some endophytic fungi of Q. gambelii could play significant ecosystem roles in litter decomposition and nutrient cycling.

Soil Properties Interacting With Microbial Metagenome in Decreasing CH4 Emission From Seasonally Flooded Marshland Following Different Stages of Afforestation.

Wetlands are the largest natural source of terrestrial CH4 emissions. Afforestation can enhance soil CH4 oxidation and decrease methanogenesis, yet the driving mechanisms leading to these effects remain unclear. We analyzed the structures of communities of methanogenic and methanotrophic microbes, quantification of mcrA and pmoA genes, the soil microbial metagenome, soil properties and CH4 fluxes in afforested and non-afforested areas in the marshland of the Yangtze River. Compared to the non-afforested land use types, net CH4 emission decreased from bare land, natural vegetation and 5-year forest plantation and transitioned to net CH4 sinks in the 10- and 20-year forest plantations. Both abundances of mcrA and pmoA genes decreased significantly with increasing plantation age. By combining random forest analysis and structural equation modeling, our results provide evidence for an important role of the abundance of functional genes related to methane production in explaining the net CH4 flux in this ecosystem. The structures of methanogenic and methanotrophic microbial communities were of lower importance as explanatory factors than functional genes in terms of in situ CH4 flux. We also found a substantial interaction between functional genes and soil properties in the control of CH4 flux, particularly soil particle size. Our study provides empirical evidence that microbial community function has more explanatory power than taxonomic microbial community structure with respect to in situ CH4 fluxes. This suggests that focusing on gene abundances obtained, e.g., through metagenomics or quantitative/digital PCR could be more effective than community profiling in predicting CH4 fluxes, and such data should be considered for ecosystem modeling.

The protection of Salicornia rubra from ultraviolet radiation by betacyanins and phenolic compounds.

Salicornia rubra is a commonly occurring annual species of the salt playas of the Great Basin Desert of the western United States. In such habitats, plants experience high levels of ultraviolet radiation, which could potentially damage DNA. As a member of the Amaranthaceae (Caryophyllales), S. rubra shoots typically contain high concentrations of the red-violet pigments called betacyanins, which are ultraviolet-absorbing compounds. Nevertheless, some specimens of S. rubra are green even when growing with full exposure to the sun. We, therefore, tested several hypotheses regarding the causes of variation among S. rubra plants in betacyanin concentration and the role of betacyanins in the absorption of ultraviolet radiation. We measured ultraviolet radiation absorption and the concentrations of betacyanins and phenolic compounds of the cell sap expressed from red and green plants growing in full sun, as well as plants grown under various levels of shade. We found that while betacyanin concentrations were predictable from plant color (red plants contained more betacyanins than green plants), the ability to absorb ultraviolet radiation was determined primarily by the concentration of phenolic compounds, which was determined by the level of exposure to the sun. Therefore, the DNA of green plants growing in full sun appears to be at no greater risk than the DNA of red plants.

Large losses of ammonium-nitrogen from a rice ecosystem under elevated CO2.

Inputs of nitrogen into terrestrial ecosystems, mainly via the use of ammonium-based fertilizers in agroecosystems, are enormous, but the fate of this nitrogen under elevated atmospheric carbon dioxide (CO2) is not well understood. We have taken advantage of a 15-year free-air CO2 enrichment study to investigate the influence of elevated CO2 on the transformation of ammonium-nitrogen in a rice ecosystem in which ammonium is usually assumed to be stable under anaerobic conditions. We demonstrate that elevated CO2 causes substantial losses of ammonium-nitrogen that result from anaerobic oxidation of ammonium coupled to reduction of iron. We identify a new autotrophic member of the bacterial order Burkholderiales that may use soil CO2 as a carbon source to couple anaerobic ammonium oxidation and iron reduction. These findings offer insight into the coupled cycles of nitrogen and iron in terrestrial ecosystems and raise questions about the loss of ammonium-nitrogen from arable soils under future climate-change scenarios.

The Dynamics of Interacting Bacterial and Fungal Communities of the Mouse Colon Following Antibiotics.

We tested two hypotheses concerning the dynamics of intestinal microbial communities of young mice following antibiotic-induced disturbance. The first is that disturbance of the bacterial community causes disturbance of the fungal community. Our results were consistent with that hypothesis. Antibiotics significantly altered bacterial community structure. Antibiotics also altered fungal community structure, significantly increasing the relative abundance of Candida lusitaniae, a known pathogen, while simultaneously significantly decreasing the relative abundances of several other common fungal species. The result was a temporary decrease in fungal diversity. Moreover, bacterial load was negatively correlated with the relative abundances of Candida lusitaniae and Candida parapsilosis, while it was positively correlated with the relative abundances of many other fungal species. Our second hypothesis is that control mice serve as a source of probiotics capable of invading intestines of mice with disturbed microbial communities and restoring pre-antibiotic bacterial and fungal communities. However, we found that control mice did not restore disturbed microbial communities. Instead, mice with disturbed microbial communities induced disturbance in control mice, consistent with the hypothesis that antibiotic-induced disturbance represents an alternate stable state that is easier to achieve than to correct. Our results indicate the occurrence of significant interactions among intestinal bacteria and fungi and suggest that the stimulation of certain bacterial groups may potentially be useful in countering the dominance of fungal pathogens such as Candida spp. However, the stability of disturbed microbial communities could complicate recovery.

The role of resource transfer in positive, non-additive litter decomposition.

Naturally occurring, mixed litter decomposes at unpredictable rates when individual components do not decompose in mixtures as they do individually. Consequently, nutrient, carbon and energy fluxes associated with decomposition may be difficult to predict. However, predictability is improved when we understand the mechanisms responsible for such non-additive decomposition. In this study, we explored mechanisms to explain our previous observation that an approximately 30% increase in oat straw decomposition due to the presence of clover litter is associated with a significant increase in the activity of cellobiohydrolase, an enzyme involved in litter decomposition. We hypothesized that resources limiting decomposer microbe enzyme activity in oat straw can be supplied by clover litter. Amendment of oat straw with water, NH4Cl, glucose, or NH4Cl combined with glucose did not account for the significant, positive effect of clover litter on oat straw decomposition and cellobiohydrolase activity. However, amendment of oat straw with a complete set of mineral nutrients for plant growth did account for the entire effect of clover litter, and the addition of the complete set of mineral nutrients without N accounted for the majority of the clover effect. In our system, therefore, the majority of the positive effect of clover litter on oat straw decomposition and cellobiohydrolase activity was unexpectedly not attributable to the transfer from clover to oat straw of labile N. We found that mineral soil could substitute for the mineral nutrients other than N. This highlights the role of soil as a potential source of limiting resources for microbes decomposing litter. It may also explain why positive, non-additive decomposition has been observed in some previous studies but not in others depending on whether the soil supplied a resource that limited decomposer activity.

The role of inoculum dispersal and plant species identity in the assembly of leaf endophytic fungal communities.

Because of disturbance and plant species loss at the local level, many arid ecosystems in the western USA benefit from revegetation. There is a growing interest in improving revegetation success by purposefully inoculating revegetation plants with mutualistic endophytic fungi that increase plant stress tolerance. However, inoculant fungi must compete against fungi that indigenous to the habitat, many of which may not be mutualistic. Our overall goal, therefore, is to learn how to efficiently colonize revegetation plants using endophytic fungal inoculum. The goal will be facilitated by understanding the factors that limit colonization of plants by endophytic fungi, including inoculum dispersal and host compatibility. We analyzed endophytic fungal communities in leaves of Bromus tectorum and Elymus elymoides (Poaceae), Chrysothamnus depressus and Artemisia tridentata (Asteraceae), Alyssum alyssoides (Brassicaceae) and Atriplex canescens (Amaranthaceae), each occurring in each of 18 field plots. We found that dispersal limitation was significant for endophytic fungal communities of Atriplex canescens and Bromus tectorum, accounting for 9 and 17%, respectively, of the variation in endophytic fungal community structure, even though the maximum distance between plots was only 350 m. Plant species identity accounted for 33% of the variation in endophytic fungal community structure. These results indicate that the communities of endophytic fungi assembling in these plant species depend significantly on proximity to inoculum source as well as the identity of the plant species. Therefore, if endophytic fungi are to be used to facilitate revegetation by these plant species, land managers may find it profitable to consider both the proximity of inoculum to revegetation plants and the suitability of the inoculum to targeted host plant species.

Biotic filtering of endophytic fungal communities in Bromus tectorum.

The assembly of horizontally transmitted endophytic fungi within plant tissues may be affected by "biotic filtering". In other words, only particular endophytic fungal taxa from the available inoculum pool may be able to colonize a given plant species. We tested that hypothesis in Bromus tectorum, an important invasive species in the arid, western United States. We collected seed from Bromus tectorum and sources of inoculum for endophytic fungi including soil and various kinds of plant litter at a field site in central Utah. We characterized, using Illumina sequencing, the endophytic fungal communities in the various inoculum sources, inoculated Bromus tectorum seedlings under gnotobiotic conditions with the various sources, and then characterized the communities of endophytic fungi that assembled in their roots and leaves. Different inoculum sources containing significantly different endophytic fungal communities produced complex communities of endophytic fungi in leaves and roots of Bromus tectorum. In leaves, the communities assembling from the various inoculum sources were not significantly different from each other and, in roots, they were only slightly different from each other, mainly due to variation in a single fungal OTU, Coprinopsis brunneofibrillosa. Consequently, there was significantly more variation in the structure of the communities of endophytic fungi among the inoculum sources than in the resultant endophytic fungal communities in the leaves and roots of Bromus tectorum. These results are consistent with biotic filtering playing a significant role in endophytic fungal community assembly.

Exploring the role of ectomycorrhizal fungi in soil carbon dynamics.

The extent to which ectomycorrhizal (ECM) fungi enable plants to access organic nitrogen (N) bound in soil organic matter (SOM) and transfer this growth-limiting nutrient to their plant host, has important implications for our understanding of plant-fungal interactions, and the cycling and storage of carbon (C) and N in terrestrial ecosystems. Empirical evidence currently supports a range of perspectives, suggesting that ECM vary in their ability to provide their host with N bound in SOM, and that this capacity can both positively and negatively influence soil C storage. To help resolve the multiplicity of observations, we gathered a group of researchers to explore the role of ECM fungi in soil C dynamics, and propose new directions that hold promise to resolve competing hypotheses and contrasting observations. In this Viewpoint, we summarize these deliberations and identify areas of inquiry that hold promise for increasing our understanding of these fundamental and widespread plant symbionts and their role in ecosystem-level biogeochemistry.

Common mycorrhizal networks influence the distribution of mineral nutrients between an invasive plant, Solidago canadensis, and a native plant, Kummerowa striata.

Invasive species often reduce ecosystem services and lead to a serious threat to native biodiversity. Roots of invasive plants are often linked to roots of native plants by common mycorrhizal networks (CMNs) of arbuscular mycorrhizal (AM) fungi, but whether and how CMNs mediate interactions between invasive and native plant species remains largely uninvestigated. We conducted two microcosm experiments, one in which we amended the soil with mineral N and another in which we amended the soil with mineral P. In each experiment, we grew a pair of test plants consisting of Kummerowia striata (native to our research site) and Solidago canadensis (an invasive species). CMNs were established between the plants, and these were either left intact or severed. Intact CMNs increased growth and nutrient acquisition by S. canadensis while they decreased nutrient acquisition by K. striata in comparison with severed CMNs. 15N and P analyses indicated that compared to severed CMNs, intact CMNs preferentially transferred mineral nutrients to S. canadensis. CMNs produced by different species of AM fungi had slightly different effects on the interaction between these two plant species. These results highlight the role of CMNs in the understanding of interactions between the invasive species S. canadensis and its native neighbor.

The continuing relevance of "older" mycorrhiza literature: insights from the work of John Laker Harley (1911-1990).

To new generations of scientists beginning their careers in research, we strongly recommend the practice of reading older literature. To illustrate the value of doing so, we highlight six insights of one of the most influential mycorrhiza researchers of the twentieth century, Jack Harley. These insights concerning mycotrophy, the new niche, the sheath, C cycling, N cycling, and mutualism were published prior to 1975 and so may have escaped the notice of many, but they laid the groundwork for some of the most important research of today.

A regionally-adapted implementation of conservation agriculture delivers rapid improvements to soil properties associated with crop yield stability.

Climate models predict increasing weather variability, with negative consequences for crop production. Conservation agriculture (CA) may enhance climate resilience by generating certain soil improvements. However, the rate at which these improvements accrue is unclear, and some evidence suggests CA can lower yields relative to conventional systems unless all three CA elements are implemented: reduced tillage, sustained soil cover, and crop rotational diversity. These cost-benefit issues are important considerations for potential adopters of CA. Given that CA can be implemented across a wide variety of regions and cropping systems, more detailed and mechanistic understanding is required on whether and how regionally-adapted CA can improve soil properties while minimizing potential negative crop yield impacts. Across four US states, we assessed short-term impacts of regionally-adapted CA systems on soil properties and explored linkages with maize and soybean yield stability. Structural equation modeling revealed increases in soil organic matter generated by cover cropping increased soil cation exchange capacity, which improved soybean yield stability. Cover cropping also enhanced maize minimum yield potential. Our results demonstrate individual CA elements can deliver rapid improvements in soil properties associated with crop yield stability, suggesting that regionally-adapted CA may play an important role in developing high-yielding, climate-resilient agricultural systems.

Mycorrhizal fungi and roots are complementary in foraging within nutrient patches.

The roots of the majority of tree species are associated with either arbuscular mycorrhizal (AM) or ectomycorrhizal (EM) fungi. The absorptive roots of tree species also vary widely in their diameter. The linkages between root thickness, mycorrhiza type and nutrient foraging are poorly understood. We conducted a large root ingrowth experiment in the field to investigate how absorptive roots of varying thickness and their associated fungi (AM vs. EM) exploit different nutrient patches (inorganic and organic) in a common garden. In nutrient-rich patches, thin-root tree species more effectively proliferated absorptive roots than thick-root tree species, whereas thick-root tree species proliferated more mycorrhizal fungal biomass than thin-root tree species. Moreover, nutrient patches enriched with organic materials resulted in greater root and mycorrhizal fungal proliferation compared to those enriched with inorganic nutrients. Irrespective of root morphology, AM tree species had higher root foraging precision than mycorrhizal hyphae foraging precision within organic patches, whereas EM tree species exhibited the opposite. Our findings that roots and mycorrhizal fungi are complementary in foraging within nutrient patches provide new insights into species coexistence and element cycling in terrestrial ecosystems.

Root morphology and mycorrhizal symbioses together shape nutrient foraging strategies of temperate trees.

Photosynthesis by leaves and acquisition of water and minerals by roots are required for plant growth, which is a key component of many ecosystem functions. Although the role of leaf functional traits in photosynthesis is generally well understood, the relationship of root functional traits to nutrient uptake is not. In particular, predictions of nutrient acquisition strategies from specific root traits are often vague. Roots of nearly all plants cooperate with mycorrhizal fungi in nutrient acquisition. Most tree species form symbioses with either arbuscular mycorrhizal (AM) or ectomycorrhizal (EM) fungi. Nutrients are distributed heterogeneously in the soil, and nutrient-rich "hotspots" can be a key source for plants. Thus, predicting the foraging strategies that enable mycorrhizal root systems to exploit these hotspots can be critical to the understanding of plant nutrition and ecosystem carbon and nutrient cycling. Here, we show that in 13 sympatric temperate tree species, when nutrient availability is patchy, thinner root species alter their foraging to exploit patches, whereas thicker root species do not. Moreover, there appear to be two distinct pathways by which thinner root tree species enhance foraging in nutrient-rich patches: AM trees produce more roots, whereas EM trees produce more mycorrhizal fungal hyphae. Our results indicate that strategies of nutrient foraging are complementary among tree species with contrasting mycorrhiza types and root morphologies, and that predictable relationships between below-ground traits and nutrient acquisition emerge only when both roots and mycorrhizal fungi are considered together.

MycoDB, a global database of plant response to mycorrhizal fungi.

Plants form belowground associations with mycorrhizal fungi in one of the most common symbioses on Earth. However, few large-scale generalizations exist for the structure and function of mycorrhizal symbioses, as the nature of this relationship varies from mutualistic to parasitic and is largely context-dependent. We announce the public release of MycoDB, a database of 4,010 studies (from 438 unique publications) to aid in multi-factor meta-analyses elucidating the ecological and evolutionary context in which mycorrhizal fungi alter plant productivity. Over 10 years with nearly 80 collaborators, we compiled data on the response of plant biomass to mycorrhizal fungal inoculation, including meta-analysis metrics and 24 additional explanatory variables that describe the biotic and abiotic context of each study. We also include phylogenetic trees for all plants and fungi in the database. To our knowledge, MycoDB is the largest ecological meta-analysis database. We aim to share these data to highlight significant gaps in mycorrhizal research and encourage synthesis to explore the ecological and evolutionary generalities that govern mycorrhizal functioning in ecosystems.