The exon junction complex controls the splicing of MAPK and other long intron-containing transcripts in Drosophila.

Signaling pathways are controlled by a vast array of posttranslational mechanisms. By contrast, little is known regarding the mechanisms that regulate the expression of their core components. We conducted an RNAi screen in Drosophila for factors modulating RAS/MAPK signaling and identified the Exon Junction Complex (EJC) as a key element of this pathway. The EJC binds the exon-exon junctions of mRNAs and thus far, has been linked exclusively to postsplicing events. Here, we report that the EJC is required for proper splicing of mapk transcripts by a mechanism that apparently controls exon definition. Moreover, whole transcriptome and RT-PCR analyses of EJC-depleted cells revealed that the splicing of long intron-containing genes, which includes mapk, is sensitive to EJC activity. These results identify a role for the EJC in the splicing of a subset of transcripts and suggest that RAS/MAPK signaling depends on the regulation of MAPK levels by the EJC.

Prairie restoration promotes the abundance and diversity of mutualistic arbuscular mycorrhizal fungi.

Predicting how biological communities assemble in restored ecosystems can assist in conservation efforts, but most research has focused on plants, with relatively little attention paid to soil microbial organisms that plants interact with. Arbuscular mycorrhizal (AM) fungi are an ecologically significant functional group of soil microbes that form mutualistic symbioses with plants and could therefore respond positively to plant community restoration. To evaluate the effects of plant community restoration on AM fungi, we compared AM fungal abundance, species richness, and community composition of five annually cultivated, conventionally managed agricultural fields with paired adjacent retired agricultural fields that had undergone prairie restoration 5-9 years prior to sampling. We hypothesized that restoration stimulates AM fungal abundance and species richness, particularly for disturbance-sensitive taxa, and that gains of new taxa would not displace AM fungal species present prior to restoration due to legacy effects. AM fungal abundance was quantified by measuring soil spore density and root colonization. AM fungal species richness and community composition were determined in soils and plant roots using DNA high-throughput sequencing. Soil spore density was 2.3 times higher in restored prairies compared to agricultural fields, but AM fungal root colonization did not differ between land use types. AM fungal species richness was 2.7 and 1.4 times higher in restored prairies versus agricultural fields for soil and roots, respectively. The abundance of Glomeraceae, a disturbance-tolerant family, decreased by 25% from agricultural to restored prairie soils but did not differ in plant roots. The abundance of Claroideoglomeraceae and Diversisporaceae, both disturbance-sensitive families, was 4.6 and 3.2 times higher in restored prairie versus agricultural soils, respectively. Species turnover was higher than expected relative to a null model, indicating that AM fungal species were gained by replacement. Our findings demonstrate that restoration can promote a relatively rapid increase in the abundance and diversity of soil microbial communities that had been degraded by decades of intensive land use, and community compositional change can be predicted by the disturbance tolerance of soil microbial taxonomic and functional groups.

Biosocial Technical Systems: An Emerging Approach to Analyse Responses to Novel Diseases.

The COVID-19 pandemic emerged in the context of a parallel epidemic of information, namely an infodemic. With the development of vaccines occurring in record time, a disinformation campaign ensued rendering the infodemic ever more troubling. As COVID-19 had to be curbed with vaccines opinion pools and surveys indicated that a minority, but relevant, part of the general public had weakened trust in public health policies and also on governmental responses to the pandemic in general. This dissent in public opinion on pandemic response is interpreted in this chapter as a controversy related to the efficacy and risks associated to vaccines. Such controversy gained momentum partly because traditional scientific communication has been largely unidirectional rather than bi-directional. We propose to apply a novel biosocial technical perspective to examine the COVID-19 pandemic controversy and communication, articulating social, biological and technical issues. The interaction between COVID-19 and vaccines, i.e. artefact-biological interactions, resulted in vaccine development in record time. However, the interaction between social systems and vaccine as artefacts was plagued by partial public reluctance in their acceptance. This rendered communication efforts ever more relevant, bringing lessons related to the importance of a more fluid bi-directional communication in future disease epidemics.

Advancing mycorrhizal trait-based ecology using both pure cultures and community-level traits.

Traits are the intermediate by which species respond to environmental filters and influence ecosystem functions. With the myriad of biogeochemical processes controlled by fungi, the past decade has witnessed a rising interest in applying trait-based approaches, core to the toolkit of plant and animal ecophysiologists, to fungi. One of the first challenges to tackle when working on fungal ecophysiology is to circumscribe the very definition of what we consider a fungal trait. Traits are characteristics/features possessed by an individual that can influence how it interacts with its environment. Here, the individual scale is both important, and problematic. Important because the very goal of comparative ecology is to measure traits on individuals belonging to known species. This allows us to populate trait databases, and syntheses of such databases can reveal key trade-offs and trait syndromes that govern species' life-histories. The scale of the individual is problematic, however, because it is hard to define for soil fungi, and because a rare minority of fungi can be sampled at the individual scale in the environment (e.g., macroscopic sporocarps, ectomycorrhizal root tips, lichen thalli). Beyond this minority, the individual organisms can only be accessed/sampled through establishing fungal cultures, which probably represents one of the main bottlenecks in the development of fungal trait databases. In this issue of Molecular Ecology, Zhang et al. (2022) show how interesting insights in fungal trait-based ecology can be gained by working at the community level.

A key role for EZH2 and associated genes in mouse and human adult T-cell acute leukemia.

In this study, we show the high frequency of spontaneous γδ T-cell leukemia (T-ALL) occurrence in mice with biallelic deletion of enhancer of zeste homolog 2 (Ezh2). Tumor cells show little residual H3K27 trimethylation marks compared with controls. EZH2 is a component of the PRC2 Polycomb group protein complex, which is associated with DNA methyltransferases. Using next-generation sequencing, we identify alteration in gene expression levels of EZH2 and acquired mutations in PRC2-associated genes (DNMT3A and JARID2) in human adult T-ALL. Together, these studies document that deregulation of EZH2 and associated genes leads to the development of mouse, and likely human, T-ALL.

Increased seedling establishment via enemy release at the upper elevational range limit of sugar maple.

The enemy release hypothesis is frequently invoked to explain invasion by nonnative species, but studies focusing on the influence of enemies on natural plant range expansion due to climate change remain scarce. We combined multiple approaches to study the influence of plant-enemy interactions on the upper elevational range limit of sugar maple (Acer saccharum) in southeastern Québec, Canada, where a previous study had demonstrated intense seed predation just beyond the range limit. Consistent with the hypothesis of release from natural enemies at the range limit, data from both natural patterns of regeneration and from seed and seedling transplant experiments showed higher seedling densities at the range edge than in the core of the species' distribution. A growth chamber experiment manipulating soil origin and temperature indicated that this so-called "happy edge" was not likely caused by temperature (i.e., the possibility that climate warming has made high elevation temperatures optimal for sugar maple) or by abiotic soil factors that vary along the elevational gradient. Finally, an insect-herbivore-exclusion experiment showed that insect herbivory was a major cause of seedling mortality in the core of sugar maple's distribution, whereas seedlings transplanted at or beyond the range edge experienced minimal herbivory (i.e., enemy release). Insect herbivory did not completely explain the high levels of seedling mortality in the core of the species' distribution, suggesting that seedlings at or beyond the range edge may also experience release from pathogens. In sum, while some effects of enemies are magnified beyond range edges (e.g., seed predation), others are dampened at and beyond the range edge (e.g., insect herbivory), such that understanding the net outcome of different biotic interactions within, at and beyond the edge of distribution is critical to predicting species' responses to global change.

Interaction type influences ecological network structure more than local abiotic conditions: evidence from endophytic and endolichenic fungi at a continental scale.

Understanding the factors that shape community assembly remains one of the most enduring and important questions in modern ecology. Network theory can reveal rules of community assembly within and across study systems and suggest novel hypotheses regarding the formation and stability of communities. However, such studies generally face the challenge of disentangling the relative influence of factors such as interaction type and environmental conditions on shaping communities and associated networks. Endophytic and endolichenic symbioses, characterized by microbial species that occur within healthy plants and lichen thalli, represent some of the most ubiquitous interactions in nature. Fungi that engage in these symbioses are hyperdiverse, often horizontally transmitted, and functionally beneficial in many cases, and they represent the diversification of multiple phylogenetic groups. We evaluated six measures of ecological network structure for >4100 isolates of endophytic and endolichenic fungi collected systematically from five sites across North America. Our comparison of these co-occurring interactions in biomes ranging from tundra to subtropical forest showed that the type of interactions (i.e., endophytic vs. endolichenic) had a much more pronounced influence on network structure than did environmental conditions. In particular, endophytic networks were less nested, less connected, and more modular than endolichenic networks in all sites. The consistency of the network structure within each interaction type, independent of site, is encouraging for current efforts devoted to gathering metadata on ecological network structure at a global scale. We discuss several mechanisms potentially responsible for such patterns and draw attention to knowledge gaps in our understanding of networks for diverse interaction types.

Navigating the labyrinth: a guide to sequence-based, community ecology of arbuscular mycorrhizal fungi.

Data generated from next generation sequencing (NGS) will soon comprise the majority of information about arbuscular mycorrhizal fungal (AMF) communities. Although these approaches give deeper insight, analysing NGS data involves decisions that can significantly affect results and conclusions. This is particularly true for AMF community studies, because much remains to be known about their basic biology and genetics. During a workshop in 2013, representatives from seven research groups using NGS for AMF community ecology gathered to discuss common challenges and directions for future research. Our goal was to improve the quality and accessibility of NGS data for the AMF research community. Discussions spanned sampling design, sample preservation, sequencing, bioinformatics and data archiving. With concrete examples we demonstrated how different approaches can significantly alter analysis outcomes. Failure to consider the consequences of these decisions may compound bias introduced at each step along the workflow. The products of these discussions have been summarized in this paper in order to serve as a guide for any researcher undertaking NGS sequencing of AMF communities.

Interplay of biotic and abiotic factors shapes tree seedling growth and root-associated microbial communities.

Root-associated microbes can alleviate plant abiotic stresses, thus potentially supporting adaptation to a changing climate or to novel environments during range expansion. While climate change is extending plant species fundamental niches northward, the distribution and colonization of mutualists (e.g., arbuscular mycorrhizal fungi) and pathogens may constrain plant growth and regeneration. Yet, the degree to which biotic and abiotic factors impact plant performance and associated microbial communities at the edge of their distribution remains unclear. Here, we use root microscopy, coupled with amplicon sequencing, to study bacterial, fungal, and mycorrhizal root-associated microbial communities from sugar maple seedlings distributed across two temperate-to-boreal elevational gradients in southern Québec, Canada. Our findings demonstrate that soil pH, soil Ca, and distance to sugar maple trees are key drivers of root-associated microbial communities, overshadowing the influence of elevation. Interestingly, changes in root fungal community composition mediate an indirect effect of soil pH on seedling growth, a pattern consistent at both sites. Overall, our findings highlight a complex role of biotic and abiotic factors in shaping tree-microbe interactions, which are in turn correlated with seedling growth. These findings have important ramifications for tree range expansion in response to shifting climatic niches.

RNA-seq analysis of 2 closely related leukemia clones that differ in their self-renewal capacity.

The molecular mechanisms regulating self-renewal of leukemia stem cells remain poorly understood. Here we report the generation of 2 closely related leukemias created through the retroviral overexpression of Meis1 and Hoxa9. Despite their apparent common origin, these clonal leukemias exhibit enormous differences in stem cell frequency (from 1 in 1.4, FLA2; to 1 in 347, FLB1), suggesting that one of these leukemias undergoes nearly unlimited self-renewal divisions. Using next-generation RNA-sequencing, we characterized the transcriptomes of these phenotypically similar, but biologically distinct, leukemias, identifying hundreds of differentially expressed genes and a large number of structural differences (eg, alternative splicing and promoter usage). Focusing on ligand-receptor pairs, we observed high expression levels of Sdf1-Cxcr4; Jagged2-Notch2/1; Osm-Gp130; Scf-cKit; and Bmp15-Tgfb1/2. Interestingly, the integrin beta 2-like gene (Itgb2l) is both highly expressed and differentially expressed between our 2 leukemias (∼ 14-fold higher in FLA2 than FLB1). In addition, gene ontology analysis indicated G-protein-coupled receptor had a much higher proportion of differential expression (22%) compared with other classes (∼ 5%), suggesting a potential role regulating subtle changes in cellular behavior. These results provide the first comprehensive transcriptome analysis of a leukemia stem cell and document an unexpected level of transcriptome variation between phenotypically similar leukemic cells.

Structure and impact of root-associated fungi in treatment wetland mesocosms.

Root fungal endophytes have been shown to play a positive role in soil phytoremediation by immobilizing or degrading contaminants. In comparison, little is known about their ecological functions and possible role in improving plant performance in treatment wetlands. In a greenhouse study, we compared the structure of fungal communities associated with Phragmites australis roots in treatment wetland mesocosms fed with pre-treated wastewater to mesocosms fed with drinking water. We evaluated the role of water source as an environmental filter structuring fungal communities, and correlated the relative abundances of fungal taxa with key services delivered by the wetlands (i.e., biomass production and nutrient removal). Mesocosms fed with wastewater had higher fungal alpha-diversity. Contrary to expectations, many fungi were unique to drinking water-fed mesocosms, suggesting that the oligotrophic conditions prevailing in these mesocosms benefited specific fungal taxa. On the other hand, wastewater-fed mesocosms had a slightly higher proportion of sequence reads belonging to fungal species recognized as potential endophytes and phytopathogens, highlighting the potential role of wastewater as a source of plant-associated fungi. Interestingly, we found contrasted association patterns between fungal species' relative abundances and different treatment wetland services (e.g., N vs P removal), such that some fungi were positively associated with N removal but negatively associated with P removal. This suggests that fungal endophytes may be functionally complementary in their contribution to distinct mesocosm services, thus supporting arguments in favor of microbial diversity in phytotechnologies. Because of the wide alpha-diversity of fungal communities, and the fact that with current databases, most species remained unassigned to a specific function (or even guild), further investigation is needed to link fungal community structure and service delivery in treatment wetlands.

Stressful, isolated, yet diverse: Green roofs have rich microbiomes that are not dominated by oligotrophic taxa.

Green roofs are unique ecosystems combining two major community assembly filters, namely stress and spatial isolation. As such, they represent an interesting model ecosystem in community ecology. In this study, we characterized the microbiome structure on 19 green roofs and 5 urban parks as a benchmark comparison (i.e. non-isolated, non-stressful habitats). Green roofs were not species depauperate, showing similar α-diversity compared to surrounding parks. We also did not find an overrepresentation of bacterial phyla typically recognized as oligotrophs, which calls into question the notion of green roofs as highly stressful habitats for bacteria, and/or the conservatism of nutritional ecophysiology at the phylum level. The geographical position of a roof, or its degree of spatial isolation (assessed through its height and area) were not important predictors of microbiome diversity and structure, suggesting that dispersal limitations impose little constraints on green roof microbiome assembly. Finally, key microbial groups (e.g. archaeal nitrifiers, Actinobacteria) were much less frequent and/or abundant on green roofs, which may have important implications for nutrient cycling and urban biogeochemistry. More work will be required to phenotype the microorganisms overrepresented on green roofs and specifically measure key soil processes in these unique urban ecosystems.

Phylogenetic structure of specialization: A new approach that integrates partner availability and phylogenetic diversity to quantify biotic specialization in ecological networks.

Biotic specialization holds information about the assembly, evolution, and stability of biological communities. Partner availabilities can play an important role in enabling species interactions, where uneven partner availabilities can bias estimates of biotic specialization when using phylogenetic diversity indices. It is therefore important to account for partner availability when characterizing biotic specialization using phylogenies. We developed an index, phylogenetic structure of specialization (PSS), that avoids bias from uneven partner availabilities by uncoupling the null models for interaction frequency and phylogenetic distance. We incorporate the deviation between observed and random interaction frequencies as weights into the calculation of partner phylogenetic α-diversity. To calculate the PSS index, we then compare observed partner phylogenetic α-diversity to a null distribution generated by randomizing phylogenetic distances among the same number of partners. PSS quantifies the phylogenetic structure (i.e., clustered, overdispersed, or random) of the partners of a focal species. We show with simulations that the PSS index is not correlated with network properties, which allows comparisons across multiple systems. We also implemented PSS on empirical networks of host-parasite, avian seed-dispersal, lichenized fungi-cyanobacteria, and hummingbird pollination interactions. Across these systems, a large proportion of taxa interact with phylogenetically random partners according to PSS, sometimes to a larger extent than detected with an existing method that does not account for partner availability. We also found that many taxa interact with phylogenetically clustered partners, while taxa with overdispersed partners were rare. We argue that species with phylogenetically overdispersed partners have often been misinterpreted as generalists when they should be considered specialists. Our results highlight the important role of randomness in shaping interaction networks, even in highly intimate symbioses, and provide a much-needed quantitative framework to assess the role that evolutionary history and symbiotic specialization play in shaping patterns of biodiversity. PSS is available as an R package at https://github.com/cjpardodelahoz/pss.

No evidence that skewing of X chromosome inactivation patterns is transmitted to offspring in humans.

Skewing of X chromosome inactivation (XCI) can occur in normal females and increases in tissues with age. The mechanisms underlying skewing in normal females, however, remain controversial. To better understand the phenomenon of XCI in nondisease states, we evaluated XCI patterns in epithelial and hematopoietic cells of over 500 healthy female mother-neonate pairs. The incidence of skewing observed in mothers was twice that observed in neonates, and in both cohorts, the incidence of XCI was lower in epithelial cells than hematopoietic cells. These results suggest that XCI incidence varies by tissue type and that age-dependent mechanisms can influence skewing in both epithelial and hematopoietic cells. In both cohorts, a correlation was identified in the direction of skewing in epithelial and hematopoietic cells, suggesting common underlying skewing mechanisms across tissues. However, there was no correlation between the XCI patterns of mothers and their respective neonates, and skewed mothers gave birth to skewed neonates at the same frequency as nonskewed mothers. Taken together, our data suggest that in humans, the XCI pattern observed at birth does not reflect a single heritable genetic locus, but rather corresponds to a complex trait determined, at least in part, by selection biases occurring after XCI.

Willow Aboveground and Belowground Traits Can Predict Phytoremediation Services.

The increasing number of contaminated sites worldwide calls for sustainable remediation, such as phytoremediation, in which plants are used to decontaminate soils. We hypothesized that better anchoring phytoremediation in plant ecophysiology has the potential to drastically improve its predictability. In this study, we explored how the community composition, diversity and coppicing of willow plantations, influenced phytoremediation services in a four-year field trial. We also evaluated how community-level plant functional traits might be used as predictors of phytoremediation services, which would be a promising avenue for plant selection in phytoremediation. We found no consistent impact of neither willow diversity nor coppicing on phytoremediation services directly. These services were rather explained by willow traits related to resource economics and management strategy along the plant "fast-slow" continuum. We also found greater belowground investments to promote plant bioconcentration and soil decontamination. These traits-services correlations were consistent for several trace elements investigated, suggesting high generalizability among contaminants. Overall, our study provides evidence, even using a short taxonomic (and thus functional) plant gradient, that traits can be used as predictors for phytoremediation efficiency for a broad variety of contaminants. This suggests that a trait-based approach has great potential to develop predictive plant selection strategies in phytoremediation trials, through a better rooting of applied sciences in fundamental plant ecophysiology.

Fresh Compost Tea Application Does Not Change Rhizosphere Soil Bacterial Community Structure, and Has No Effects on Soybean Growth or Yield.

Soil bacteria drive key ecosystem functions, including nutrient mobilization, soil aggregation and crop bioprotection against pathogens. Bacterial diversity is thus considered a key component of soil health. Conventional agriculture reduces bacterial diversity in many ways. Compost tea has been suggested as a bioinoculant that may restore bacterial community diversity and promote crop performance under conventional agriculture. Here, we conducted a field experiment to test this hypothesis in a soybean-maize rotation. Compost tea application had no influence on bacterial diversity or community structure. Plant growth and yield were also unresponsive to compost tea application. Combined, our results suggest that our compost tea bacteria did not thrive in the soil, and that the positive impacts of compost tea applications reported elsewhere may be caused by different microbial groups (e.g., fungi, protists and nematodes) or by abiotic effects on soil (e.g., contribution of nutrients and dissolved organic matter). Further investigations are needed to elucidate the mechanisms through which compost tea influences crop performance.

Bioaugmentation of treatment wetlands - A review.

Bioaugmentation in the form of artificial mycorrhization of plant roots and bacterial inoculation has been successfully implemented in several fields including soil remediation or activated sludge treatment. Likewise, bioaugmentation seems a promising approach to improve the functioning of treatment wetlands, considering that natural mycorrhization has been detected in treatment wetlands and that bacteria are the main driver of contaminant degradation processes. However, to date, full scale implementation seems to be rare. This review synthesizes the effects of bioaugmentation on different types of treatment wetlands, to a large extent performed on a microcosm (<0.5 m2) or mesocosm scale (0.51 to 5 m2). While inoculation with arbuscular mycorrhizal fungi tended to show a positive effect on the growth of some wetland plants (e.g. Phragmites australis), the mechanisms underlying such positive effects are not well understood and the effects of upscaling to full scale treatment wetlands remain unknown. Bacterial inoculation tended to promote plant growth and pollutant degradation, but longer term data is required.

Host identity influences nuclear dynamics in arbuscular mycorrhizal fungi.

The arbuscular mycorrhizal fungi (AMF) are involved in one of the most ecologically important symbioses on the planet, occurring within the roots of most land plants.1 Knowledge of even basic elements of AM fungal biology is still poor, with the discovery that AMF may in fact have a sexual life cycle being only very recently reported.2-5 AMF produce asexual spores that contain up to several thousand individual haploid nuclei6 of either largely uniform genotypes (AMF homokaryons) or nuclei originating from two parental genotypes2-5 (AMF dikaryons or heterokaryons). In contrast to the sexual dikaryons in the phyla Ascomycota and Basidiomycota,7,8 in which pairs of nuclei coexist in single hyphal compartments, AMF dikaryons carry several thousand nuclei in a coenocytic mycelium. Here, we set out to better understand the dynamics of this unique multinucleate condition by combining molecular analyses with advanced microscopy and modeling. Herein, we report that select AMF dikaryotic strains carry the distinct nucleotypes in equal proportions to one another, whereas others show an unequal distribution of parental nucleotypes. In both cases, the relative proportions within a given strain are inherently stable. Simulation models suggest that AMF dikaryons may be maintained through nuclear cooperation dynamics. Remarkably, we report that these nuclear ratios shift dramatically in response to plant host identity, revealing a previously unknown layer of genetic complexity and dynamism within the intimate interactions that occur between the partners of a prominent terrestrial symbiosis.