Microbial communities were studied in redoximorphic microsites of highly heterogeneous Gleysol at a mm scale using 16S and 18S amplicon sequencing to demonstrate if the composition of soil microbes reflects the differences in ferric and ferrous micro‐sites. In both explored gley horizons with redoximorphic features (Bg2 and Cg), ferric mottles were significantly enriched with total P and Fe and depleted of O, Si, Al, K and Ca compared with the adjacent ferrous groundmass (SEM–EDS). Ferric mottles were determined as Fe oxide coatings and hypocoatings. In Bg2, both prokaryotic and micro‐eukaryotic communities differed significantly between mottles and groundmass in composition of operational taxonomic units (OTUs) and in proportions of phyla, reflecting heterogeneities in the soil properties there. Mottles in Bg2 were characterized by increased proportion of Proteobacteria, decreased proportion of Acidobacteriota among prokaryotes and by dominance of a single proteobacterial OTU from Anaplasmataceae compared to all other samples. The composition of micro‐eukaryotes showed an opposite trend, as micro‐eukaryotes of Bg2 groundmass were unique among the other horizons, while micro‐eukaryotes of Bg2 mottles had similar composition to neighbouring horizons. Microbial communities of adjacent samples were not more similar to each other than communities of randomly selected ones in Bg2 horizon. That suggests that at mm scale, the sample distance does not represent the driving factor of microbial community composition and that the adjacent samples differ rather due to physicochemical factors. The spatial organization of microbial communities revealed in Bg2 has not reappeared in similarly organized Cg horizon, probably due to other overriding factors. The differences revealed between Bg2 and Cg horizons, including granulometric composition, content of crystalline Fe, exchangeable Al, and organic carbon, as well as exposition to groundwater, were discussed as possible reasons of the distinct organization in Cg. The similarity of pro−/eukaryotic communities of adjacent and non‐adjacent couples suggests no distance decay pattern at a mm scale. The agreement between patchiness in soil properties and microbial communities was revealed for the first time and confirms the importance of microscale patterns in soil.
Actinobacteria are important cave inhabitants, but knowledge of how anthropization and anthropization-related visual marks affect this community on cave walls is lacking. We compared Actinobacteria communities among four French limestone caves (Mouflon, Reille, Rouffignac, Lascaux) ranging from pristine to anthropized, and within Lascaux Cave between marked (wall visual marks) and unmarked areas in different rooms (Sas-1, Passage, Apse, Diaclase). In addition to the 16S rRNA gene marker, 441 bp fragments of the hsp65 gene were used and an hsp65-related taxonomic database was constructed for identification of Actinobacteria to the species level by Illumina-MiSeq analysis. The hsp65 marker revealed higher resolution for species and higher richness (99% OTU cutoff) versus 16S rRNA gene; however, more taxa were identified at higher taxonomic ranks. Actinobacteria communities varied between Mouflon and Reille caves (both pristine), and Rouffignac and Lascaux (both anthropized). Rouffignac displayed high diversity of Nocardia, suggesting human inputs, and Lascaux exhibited high Mycobacterium relative abundance, whereas Gaiellales were typical in pristine caves and the Diaclase (least affected area of Lascaux Cave). Within Lascaux, Pseudonocardiaceae dominated on unmarked walls and Streptomycetaceae (especially Streptomyces mirabilis) on marked walls, indicating a possible role in mark formation. A new taxonomic database (https://zenodo.org/record/5576074) was developed. Although not all Actinobacteria species were represented, the use of the hsp65 marker enabled species-level variations of the Actinobacteria community to be documented based on the extent of anthropogenic pressure. This approach proved effective when comparing different limestone caves or specific conditions within one cave.
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