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Abstract The potential promise of the microbiome to ameliorate a wide range of societal and ecological challenges, from disease prevention and treatment to the restoration of entire ecosystems, hinges not only on microbiome engineering but also on the stability of beneficial microbiomes. Yet the properties of microbiome stability remain elusive and challenging to discern due to the complexity of interactions and often intractable diversity within these communities of bacteria, archaea, fungi, and other microeukaryotes. Networks are powerful tools for the study of complex microbiomes, with the potential to elucidate structural patterns of stable communities and generate testable hypotheses for experimental validation. However, the implementation of these analyses introduces a cascade of dichotomies and decision trees due to the lack of consensus on best practices. Here, we provide a road map for network-based microbiome studies with an emphasis on discerning properties of stability. We identify important considerations for data preparation, network construction, and interpretation of network properties. We also highlight remaining limitations and outstanding needs for this field. This review also serves to clarify the varying schools of thought on the application of network theory for microbiome studies and to identify practices that enhance the reproducibility and validity of future work. 

Abstract PremiseThe ability of plants to adapt or acclimate to climate change is inherently linked to their interactions with symbiotic microbes, notably fungi. However, it is unclear whether fungal symbionts from different climates have different impacts on the outcome of plant–fungal interactions, especially under environmental stress. MethodsWe tested three provenances of fungal inoculum (originating from dry, moderate or wet environments) with one host plant genotype exposed to three soil moisture regimes (low, moderate and high). Inoculated and uninoculated plants were grown in controlled conditions for 151 days, then shoot and root biomass were weighed and fungal diversity and community composition determined via amplicon sequencing. ResultsThe source of inoculum and water regime elicited significant changes in plant resource allocation to shoots versus roots, but only specific inocula affected total plant biomass. Shoot biomass increased in the high water treatment but was negatively impacted by all inoculum treatments relative to the controls. The opposite was true for roots, where the low water treatment led to greater proportional root biomass, and plants inoculated with wet site fungi allocated significantly more resources to root growth than dry‐ or moderate‐site inoculated plants and the controls. Fungal communities of shoots and roots partitioned by inoculum source, water treatment, and the interaction of the two. ConclusionsThe provenance of fungi can significantly affect total plant biomass and resource allocation above‐ and belowground, with fungi derived from more extreme environments eliciting the strongest plant responses. 

Microbes are found in nearly every habitat and organism on the planet, where they are critical to host health, fitness, and metabolism. In most organisms, few microbes are inherited at birth; instead, acquiring microbiomes generally involves complicated interactions between the environment, hosts, and symbionts. Despite the criticality of microbiome acquisition, we know little about where hosts’ microbes reside when not in or on hosts of interest. Because microbes span a continuum ranging from generalists associating with multiple hosts and habitats to specialists with narrower host ranges, identifying potential sources of microbial diversity that can contribute to the microbiomes of unrelated hosts is a gap in our understanding of microbiome assembly. Microbial dispersal attenuates with distance, so identifying sources and sinks requires data from microbiomes that are contemporary and near enough for potential microbial transmission. Here, we characterize microbiomes across adjacent terrestrial and aquatic hosts and habitats throughout an entire watershed, showing that the most species-poor microbiomes are partial subsets of the most species-rich and that microbiomes of plants and animals are nested within those of their environments. Furthermore, we show that the host and habitat range of a microbe within a single ecosystem predicts its global distribution, a relationship with implications for global microbial assembly processes. Thus, the tendency for microbes to occupy multiple habitats and unrelated hosts enables persistent microbiomes, even when host populations are disjunct. Our whole-watershed census demonstrates how a nested distribution of microbes, following the trophic hierarchies of hosts, can shape microbial acquisition.