An updated LSU database and pipeline for environmental DNA identification of arbuscular mycorrhizal fungi.

Recent work established a backbone reference tree and phylogenetic placement pipeline for identification of arbuscular mycorrhizal fungal (AMF) large subunit (LSU) rDNA environmental sequences. Our previously published pipeline allowed any environmental sequence to be identified as putative AMF or within one of the major families. Despite this contribution, difficulties in implementation of the pipeline remain. Here, we present an updated database and pipeline with (1) an expanded backbone tree to include four newly described genera and (2) several changes to improve ease and consistency of implementation. In particular, packages required for the pipeline are now installed as a single folder (conda environment) and the pipeline has been tested across three university computing clusters. This updated backbone tree and pipeline will enable broadened adoption by the community, advancing our understanding of these ubiquitous and ecologically important fungi.

Composition and seasonal variation of the arbuscular mycorrhizal fungi spore community in litter, root mat, and soil from a subtropical rain forest.

Community composition and seasonal variation of sporulation of arbuscular mycorrhizal fungi (AMF) have been studied in soils from many ecosystems including subtropical forest. Yet, AMF community composition has been surveyed only from the mineral soil but not from the litter layer and the root mat, and long-term variation in sporulation is not fully understood. We sampled a 75-m2 plot from a subtropical forest to determine AMF community composition in the following habitats: the litter layer, the root mat, and the mineral soil. Moreover, samples were taken in fall, winter, spring, and summer over a 2-year period to follow the seasonal variation of AMF sporulation. We detected 47 AMF species belonging to six families and 14 genera, Glomeraceae and Acaulosporaceae being the most represented families. Sixteen species were common to all three habitats, five species were shared between two habitats, and 26 species were recovered exclusively from single habitats. While species richness was not significantly different among habitats, AMF total spore numbers were significantly higher in the litter and root mat compared to the soil. PERMANOVA did not detect a significant effect of habitats on community composition when species presence/absence was considered, but significant differences between litter versus soil and root mat versus soil were detected when spore abundance was considered. A seasonal pattern of spore abundance for species was not observed over the 2-year sampling period regardless of habitat. This study revealed that (i) different AMF species sporulate in the different habitats; thus, field surveys considering only the mineral soil might underestimate species richness and (ii) AMF species sporulate asynchronously in subtropical forest.

Environmental identification of arbuscular mycorrhizal fungi using the LSU rDNA gene region: an expanded database and improved pipeline.

Arbuscular mycorrhizal fungi (AMF; Glomeromycota) are difficult to culture; therefore, establishing a robust amplicon-based approach to taxa identification is imperative to describe AMF diversity. Further, due to low and biased sampling of AMF taxa, molecular databases do not represent the breadth of AMF diversity, making database matching approaches suboptimal. Therefore, a full description of AMF diversity requires a tool to determine sequence-based placement in the Glomeromycota clade. Nonetheless, commonly used gene regions, including the SSU and ITS, do not enable reliable phylogenetic placement. Here, we present an improved database and pipeline for the phylogenetic determination of AMF using amplicons from the large subunit (LSU) rRNA gene. We improve our database and backbone tree by including additional outgroup sequences. We also improve an existing bioinformatics pipeline by aligning forward and reverse reads separately, using a universal alignment for all tree building, and implementing a BLAST screening prior to tree building to remove non-homologous sequences. Finally, we present a script to extract AMF belonging to 11 major families as well as an amplicon sequencing variant (ASV) version of our pipeline. We test the utility of the pipeline by testing the placement of known AMF, known non-AMF, and Acaulospora sp. spore sequences. This work represents the most comprehensive database and pipeline for phylogenetic placement of AMF LSU amplicon sequences within the Glomeromycota clade.

Perennial, but not annual legumes synergistically benefit from infection with arbuscular mycorrhizal fungi and rhizobia: a meta-analysis.

Many plant species simultaneously interact with multiple symbionts, which can, but do not always, generate synergistic benefits for their host. We ask if plant life history (i.e. annual vs perennial) can play an important role in the outcomes of the tripartite symbiosis of legumes, arbuscular mycorrhizal fungi (AMF), and rhizobia. We performed a meta-analysis of 88 studies examining outcomes of legume-AMF-rhizobia interactions on plant and microbial growth. Perennial legumes associating with AMF and rhizobia grew larger than expected based on their response to either symbiont alone (i.e. their response to co-inoculation was synergistic). By contrast, annual legume growth with co-inoculation did not differ from additive expectations. AMF and rhizobia differentially increased phosphorus (P) and nitrogen (N) tissue concentration. Rhizobium nodulation increased with mycorrhizal fungi inoculation, but mycorrhizal fungi colonization did not increase with rhizobium inoculation. Microbial responses to co-infection were significantly correlated with synergisms in plant growth. Our work supports a balanced plant stoichiometry mechanism for synergistic benefits. We find that synergisms are in part driven by reinvestment in complementary symbionts, and that time-lags in realizing benefits of reinvestment may limit synergisms in annuals. Optimization of microbiome composition to maximize synergisms may be critical to productivity, particularly for perennial legumes.

Celebrating INVAM: 35 years of the largest living culture collection of arbuscular mycorrhizal fungi.

The International Culture Collection of (Vesicular-) Arbuscular Mycorrhizal Fungi-INVAM-the largest living culture collection of arbuscular mycorrhizal fungi (AMF) celebrated its 35th year in 2020. The authors record here the mission and goals of INVAM, its contribution as a living culture collection, some historical aspects of INVAM, and describe the advances in mycorrhizology and AMF systematics after INVAM moved to West Virginia University. This commentary emphasizes the importance of a living culture collection to preserve germplasm and to educate and assist researchers in mycorrhizal science.

Biogeography of arbuscular mycorrhizal fungi (Glomeromycota): a phylogenetic perspective on species distribution patterns.

Information on the biogeography of arbuscular mycorrhizal fungi (AMF) is important because this group of obligately symbiotic soil microbes is a ubiquitous and functionally critical component of terrestrial ecosystems. In this paper, we utilize a biogeography database summarizing data on AMF species distribution linked to geographic and environmental conditions to describe global distribution patterns and interpret these patterns within a phylogenetic perspective. The data were obtained from accessions in living culture collections (INVAM, CICG), species descriptions, and other published literature from 1960 to 2012. The database contains 7105 records, 6396 of them from 768 published papers and the remaining 709 from culture accessions. Glomeromycotan species were recorded in all seven continents, 87 countries, 11 biogeographical realms, and 14 biomes. The distribution of families differed among climatic zones and continents, but they, together with all genera, appear to be cosmopolitan. Distribution of AMF species shows a slight decrease from low to high latitudes, but this decrease is steeper in the southern than in the northern hemisphere. A total of 189 species is shared between ancient supercontinents Gondwana and Laurasia and 78 species are common to all climatic zones. Ninety-five species (43% of the total) have known cosmopolitan distribution, including members of all genera except Redeckera. Some species have disjunct distribution and 26% of species have been registered from only one continent. Data on AMF distribution challenge the "Everything is everywhere" hypothesis in favor of the "moderate endemicity model" for species distribution. Data from this study provide a foundation to formulate and test hypotheses of biogeographic patterns and processes in Glomeromycota.

Effectiveness of Arbuscular Mycorrhizal Fungal Isolates from the Land Uses of Amazon Region in Symbiosis with Cowpea

ABSTRACT Arbuscular mycorrhizal fungi provide several ecosystem services, including increase in plant growth and nutrition. The occurrence, richness, and structure of arbuscular mycorrhizal fungi communities are influenced by human activities, which may affect the functional benefits of these components of the soil biota. In this study, 13 arbuscular mycorrhizal fungi isolates originating from soils with different land uses in the Alto Solimões-Amazon region were evaluated regarding their effect on growth, nutrition, and cowpea yield in controlled conditions using two soils. Comparisons with reference isolates and a mixture of isolates were also performed. Fungal isolates exhibited a wide variability associated with colonization, sporulation, production of aboveground biomass, nitrogen and phosphorus uptake, and grain yield, indicating high functional diversity within and among fungal species. A generalized effect of isolates in promoting phosphorus uptake, increase in biomass, and cowpea yield was observed in both soils. The isolates of Glomus were the most efficient and are promising isolates for practical inoculation programs. No relationship was found between the origin of fungal isolate (i.e. land use) and their symbiotic performance in cowpea.

Effectiveness of Arbuscular Mycorrhizal Fungal Isolates from the Land Uses of Amazon Region in Symbiosis with Cowpea.

Arbuscular mycorrhizal fungi provide several ecosystem services, including increase in plant growth and nutrition. The occurrence, richness, and structure of arbuscular mycorrhizal fungi communities are influenced by human activities, which may affect the functional benefits of these components of the soil biota. In this study, 13 arbuscular mycorrhizal fungi isolates originating from soils with different land uses in the Alto Solimões-Amazon region were evaluated regarding their effect on growth, nutrition, and cowpea yield in controlled conditions using two soils. Comparisons with reference isolates and a mixture of isolates were also performed. Fungal isolates exhibited a wide variability associated with colonization, sporulation, production of aboveground biomass, nitrogen and phosphorus uptake, and grain yield, indicating high functional diversity within and among fungal species. A generalized effect of isolates in promoting phosphorus uptake, increase in biomass, and cowpea yield was observed in both soils. The isolates of Glomus were the most efficient and are promising isolates for practical inoculation programs. No relationship was found between the origin of fungal isolate (i.e. land use) and their symbiotic performance in cowpea.

An evidence-based consensus for the classification of arbuscular mycorrhizal fungi (Glomeromycota).

The publication of a large number of taxon names at all levels within the arbuscular mycorrhizal fungi (Glomeromycota) has resulted in conflicting systematic schemes and generated considerable confusion among biologists working with these important plant symbionts. A group of biologists with more than a century of collective experience in the systematics of Glomeromycota examined all available molecular-phylogenetic evidence within the framework of phylogenetic hypotheses, incorporating morphological characters when they were congruent. This study is the outcome, wherein the classification of Glomeromycota is revised by rejecting some new names on the grounds that they are founded in error and by synonymizing others that, while validly published, are not evidence-based. The proposed "consensus" will provide a framework for additional original research aimed at clarifying the evolutionary history of this important group of symbiotic fungi.

Dosage-dependent shift in the spore community of arbuscular mycorrhizal fungi following application of tannery sludge.

The controlled disposal of tannery sludge in agricultural soils is a viable alternative for recycling such waste; however, the impact of this practice on the arbuscular mycorrhizal fungi (AMF) communities is not well understood. We studied the effects of low-chromium tannery sludge amendment in soils on AMF spore density, species richness and diversity, and root colonization levels. Sludge was applied at four doses to an agricultural field in Rolândia, Paraná state, Brazil. The sludge was left undisturbed on the soil surface and then the area was harrowed and planted with corn. The soil was sampled at four intervals and corn roots once within a year (2007/2008). AMF spore density was low (1 to 49 spores per 50 cm(3) of soil) and decreased as doses of tannery sludge increased. AMF root colonization was high (64%) and unaffected by tannery sludge. Eighteen AMF species belonging to six genera (Acaulospora, Glomus, Gigaspora, Scutellospora, Paraglomus, and Ambispora) were recorded. At the sludge doses of 9.0 and 22.6 Mg ha(-1), we observed a decrease in AMF species richness and diversity, and changes in their relative frequencies. Hierarchical grouping analysis showed that adding tannery waste to the soil altered AMF spore community in relation to the control, modifying the mycorrhizal status of soil and selectively favoring the sporulation of certain species.