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Larger legume plants host a greater diversity of symbiotic nitrogen-fixing bacteria

A major goal in microbial ecology is to understand the factors that structure bacterial communities across space and time. For microbes that have symbiotic relationships with plants, an important factor that may influence their communities is host size or age, yet this has received little attention. Using tree diameter size as a proxy for age, we quantified the diversity of rhizobia that associate with an endemic legume, Acacia acuminata, of variable size across a climate gradient in southwest Australia. We examined the 16S rRNA diversity (V1-V3 hypervariable region) of rhizobia at the taxonomic level and at higher sequence level diversity within taxonomic groups. We identified 3 major taxonomic clades that associated with Acacia acuminata: Bradyrhizobiaceae, Rhizobiaceae, and Burkholderiaceae. Within these groups, we found extensive genetic variability, especially within Bradyrhizobiaceae. Using binomial multivariate statistical models that controlled for other factors that affect plant size and rhizobia community structure (climate and local soil characteristics), we determined that soil sampled at the base of larger Acacia trees was much more likely to contain a greater number of taxonomic clades and cryptic genetic variants within the Rhizobiaceae clade. Despite strong influences of climate and highly heterogeneous soil conditions on rhizobial diversity, our results show that host tree size is a prominent factor in structuring nitrogen-fixing symbionts diversity across a large landscape. The identification of a positive relationship between plant host size and microbial diversity raise interesting questions about the role of plant host size in driving ecological processes that govern microbial community assembly. Specifically, our results suggest that hosts may modify the habitat of their surrounding soil to enhance growth (niche construction hypothesis) or that symbiotic microbes have large differences in dispersal capability. Our results also suggest that host plants may be analogous to ‘islands’, where larger legume hosts may accumulate diversity over time, through migration opportunities or in situ diversification. From a practical perspective, including plant size as an additional variable may assist sampling and analyses designs of future soil microbial studies.