Stéphane Uroz

Mineral weathering by bacteria: ecology, actors and mechanisms

Soil microbes play an essential role in the environment by contributing to the release of key nutrients from primary minerals that are required not only for their own nutrition but also for that of plants. Although the role of fungi in mineral weathering is beginning to be elucidated, the relative impact of bacteria in this process and the molecular mechanisms involved remain poorly understood. Here, we discuss the ecological relevance of bacterial weathering, mainly in the soil and especially in acidic forest ecosystems, which strongly depend on mineral weathering for their sustainability. We also present highlights from recent studies showing molecular mechanisms and genetic determinants involved in the dissolution of complex minerals under aerobic conditions. Finally, we consider the potential applications of genomic resources to the study of bacterial weathering.

Pyrosequencing reveals a contrasted bacterial diversity between oak rhizosphere and surrounding soil.

Several reports have highlighted that forest soil samples are more phylum-rich than agricultural soil samples. However, little is known about the structure and richness of the bacterial communities in forest soil. Using high-throughput next generation 454 pyrosequencing, we deeply investigated the diversity of bacterial communities colonizing the oak rhizosphere niche and the surrounding soil. From three spatially independent soil samples, we obtained over 300 000 partial 16S rRNA gene sequences. The most abundant bacterial groups were the Acidobacteria, Proteobacteria and unclassified bacteria. Multifactorial analysis of the relative proportions of the different phyla revealed a net differentiation of the bacterial communities present in the rhizosphere and soil environments, suggesting an oak rhizosphere effect. Significantly more β-, γ- and unclassified Proteobacteria inhabited the rhizosphere when compared with the surrounding soil. Conversely, significantly more unclassified bacteria were detected in the bulk soil than in the rhizosphere, demonstrating that the soil remains a challenging reservoir of complexity. This work increases our understanding of the niche effect on bacterial diversity and on the rare phylogenetic groups inhabiting the soil.

Effect of the mycorrhizosphere on the genotypic and metabolic diversity of the bacterial communities involved in mineral weathering in a forest soil.

ABSTRACT To date, several bacterial species have been described as mineral-weathering agents which improve plant nutrition and growth. However, the possible relationships between mineral-weathering potential, taxonomic identity, and metabolic ability have not been investigated thus far. In this study, we characterized a collection of 61 bacterial strains isolated from Scleroderma citrinum mycorrhizae, the mycorrhizosphere, and the adjacent bulk soil in an oak forest. The ability of bacteria to weather biotite was assessed with a new microplate bioassay that measures the pH and the quantity of iron released from this mineral. We showed that weathering bacteria occurred more frequently in the vicinity of S. citrinum than in the bulk soil. Moreover, the weathering efficacy of the mycorrhizosphere bacterial isolates was significantly greater than that of the bulk soil isolates. All the bacterial isolates were identified by partial 16S rRNA gene sequence analysis as members of the genera Burkholderia, Collimonas, Pseudomonas, and Sphingomonas, and their carbon metabolism was characterized by the BIOLOG method. The most efficient isolates belonged to the genera Burkholderia and Collimonas. Multivariate analysis resulted in identification of three metabolic groups, one of which contained mainly bacterial isolates associated with S. citrinum and exhibiting high mineral-weathering potential. Therefore, our results support the hypothesis that by its carbon metabolism this fungus selects in the bulk soil reservoir a bacterial community with high weathering potential, and they also address the question of functional complementation between mycorrhizal fungi and bacteria in the ectomycorrhizal complex for the promotion of tree nutrition.

Soil networks become more connected and take up more carbon as nature restoration progresses

Soil organisms have an important role in aboveground community dynamics and ecosystem functioning in terrestrial ecosystems. However, most studies have considered soil biota as a black box or focussed on specific groups, whereas little is known about entire soil networks. Here we show that during the course of nature restoration on abandoned arable land a compositional shift in soil biota, preceded by tightening of the belowground networks, corresponds with enhanced efficiency of carbon uptake. In mid- and long-term abandoned field soil, carbon uptake by fungi increases without an increase in fungal biomass or shift in bacterial-to-fungal ratio. The implication of our findings is that during nature restoration the efficiency of nutrient cycling and carbon uptake can increase by a shift in fungal composition and/or fungal activity. Therefore, we propose that relationships between soil food web structure and carbon cycling in soils need to be reconsidered.

The bacterial genus Collimonas mycophagy, weathering, and other adaptive solutions to life in oligotrophic soil environments

This minireview provides a synopsis of past and present research on the biology and ecology of members of the bacterial genus Collimonas. From the distribution, abundance and functional behaviours of these so-called collimonads emerges a general picture of bacterial adaptation to low-nutrient soil environments. Among these adaptations is the ability to extract nutrients from living fungi (mycophagy) and from rocks and minerals (weathering). This unique combination of properties will be discussed in the context of other interactions that collimonads have with their biotic and abiotic surroundings, such as the ability to inhibit fungal growth (fungistasis), protect plant roots from fungal disease (biocontrol), and degrade natural polymers and synthetic pollutants (biodegradation). Future research on Collimonas is expected to take advantage of the genomic tools and resources that are becoming available to uncover and describe the genes and gene functions that distinguish this group of bacteria and are the basis for its phenotypes. Potential applications of collimonads include the control of unwanted fungi, for example in agriculture, their use as biological indicators of soil quality and fertility, and as a source of bioactive compounds.

Functional Assays and Metagenomic Analyses Reveals Differences between the Microbial Communities Inhabiting the Soil Horizons of a Norway Spruce Plantation

In temperate ecosystems, acidic forest soils are among the most nutrient-poor terrestrial environments. In this context, the long-term differentiation of the forest soils into horizons may impact the assembly and the functions of the soil microbial communities. To gain a more comprehensive understanding of the ecology and functional potentials of these microbial communities, a suite of analyses including comparative metagenomics was applied on independent soil samples from a spruce plantation (Breuil-Chenue, France). The objectives were to assess whether the decreasing nutrient bioavailability and pH variations that naturally occurs between the organic and mineral horizons affects the soil microbial functional biodiversity. The 14 Gbp of pyrosequencing and Illumina sequences generated in this study revealed complex microbial communities dominated by bacteria. Detailed analyses showed that the organic soil horizon was significantly enriched in sequences related to Bacteria, Chordata, Arthropoda and Ascomycota. On the contrary the mineral horizon was significantly enriched in sequences related to Archaea. Our analyses also highlighted that the microbial communities inhabiting the two soil horizons differed significantly in their functional potentials according to functional assays and MG-RAST analyses, suggesting a functional specialisation of these microbial communities. Consistent with this specialisation, our shotgun metagenomic approach revealed a significant increase in the relative abundance of sequences related glycoside hydrolases in the organic horizon compared to the mineral horizon that was significantly enriched in glycoside transferases. This functional stratification according to the soil horizon was also confirmed by a significant correlation between the functional assays performed in this study and the functional metagenomic analyses. Together, our results suggest that the soil stratification and particularly the soil resource availability impact the functional diversity and to a lesser extent the taxonomic diversity of the bacterial communities.

Degradation of N-acyl homoserine lactone quorum sensing signal molecules by forest root-associated fungi

A collection of mycorrhizal and nonmycorrhizal root-associated fungi coming from forest environments was screened for their ability to degrade N-acyl homoserine lactones (AHL) or to prevent AHL recognition by producing quorum sensing inhibitors (QSI). No production of QS-inhibitors or -activators was detected using the two biosensors Chromobacterium violaceum CV026 and Agrobacterium tumefaciens in the culture supernatant of these fungi. However, the ability to degrade C6- and 3O,C6-HSL was detected for three fungal isolates. Acidification assay revealed that the AHL were degraded by a lactonase activity for two of these isolates. These results demonstrated for the first time that the forest root-associated fungi are capable of degrading the AHL signal molecules.

Black truffle-associated bacterial communities during the development and maturation of Tuber melanosporum ascocarps and putative functional roles

Although truffles are cultivated since decades, their life cycle and the conditions stimulating ascocarp formation still remain mysterious. A role for bacteria in the development of several truffle species has been suggested but few is known regarding the natural bacterial communities of Périgord Black truffle. Thus, the aim of this study was to decipher the structure and the functional potential of the bacterial communities associated to the Black truffle in the course of its life cycle and along truffle maturation. A polyphasic approach combining 454-pyrosequencing of 16S rRNA gene, TTGE, in situ hybridization and functional GeoChip 3.0 revealed that Black truffle ascocarps provide a habitat to complex bacterial communities that are clearly differentiated from those of the surrounding soil and the ectomycorrhizosphere. The composition of these communities is dynamic and evolves during the maturation of the ascocarps with an enrichment of specific taxa and a differentiation of the gleba and peridium-associated bacterial communities. Genes related to nitrogen and sulphur cycling were enriched in the ascocarps. Together, these data paint a new picture of the interactions existing between truffle and bacteria and of the potential role of these bacteria in truffle maturation.

Ancestral alliances: Plant mutualistic symbioses with fungi and bacteria

Taking a look at plant-microbe relationships Ever since plants colonized land, they have evolved a range of mutualistic associations with bacteria and fungi. Indeed, such associations were probably required for plants to grow on harsh, nutrient-poor surfaces. Martin et al. review the spectrum of plant-microbe symbioses and their evolution, including evidence from the Rhynie Chert of the Devonian period and modern associations. Surprisingly, diverse functional plant-microbial symbioses have several common conserved features, including signaling pathways, immune evasion, and root development. Science, this issue p. eaad4501 BACKGROUND Among the extensive cortège of plant-associated microorganisms (the so-called plant microbiota), mutualistic fungal and bacterial symbionts are striking examples of soil microorganisms that have successfully coevolved with their hosts since plants adapted to terrestrial ecosystems. They promote plant growth by facilitating the acquisition of scarce nutrients. In these associations, plant root colonization requires complex molecular cross-talk between symbiotic partners to activate a variety of host developmental pathways and specialized symbiotic tissues and organs. Despite the evolutionary distances that separate mycorrhizal and nitrogen-fixing symbioses, recent research has identified certain highly conserved features associated with early stages of root colonization. We focus on recent and emerging areas of investigation concerning these major mutualistic symbioses and discuss some of the molecular pathways and cellular mechanisms involved in their evolution and development. ADVANCES Phylogenomic analyses and divergence time estimates based on symbiotic plant fossils are shedding light on the evolution of mutualistic symbioses. The earliest land plants [~407 million years ago (Ma)] were associated with fungi producing mycorrhiza-like intracellular structures similar to extant symbioses involving Glomeromycotina and Mucoromycotina. Arbuscular mycorrhizal endosymbioses then diversified by the Late Carboniferous. Pinaceae species from the Late Jurassic and Early Cretaceous (~180 Ma) formed the first ectomycorrhizal associations involving Dikarya. More recently, certain angiosperms evolved a “predisposition” for the evolution of nitrogen-fixing root nodule symbioses (~100 Ma) with bacteria. A conserved core module of the “common symbiotic signaling pathway” (CSSP) is shared by all host plants that establish endosymbioses, including arbuscular mycorrhizal, rhizobial, and actinorhizal associations. This striking conservation among widely divergent host species underlines the shared evolutionary origin for this ancient symbiotic signaling pathway. Furthermore, chitin-based signaling molecules secreted by both arbuscular mycorrhizal fungi and rhizobia activate the host CSSP after perception by related receptor-like kinases. Downstream signal transduction pathways then lead to the apoplastic intracellular infection modes that characterize the majority of these associations and, finally, to the coordinated development of sophisticated bidirectional symbiotic interfaces found in both arbuscules and nitrogen-fixing nodules. A common feature of all these mutualistic associations is phytohormone-associated modifications of root development, which lead to an increase in potential colonization sites as well as major structural and functional changes to the root during the establishment of symbiotic tissues. OUTLOOK Although we are at last beginning to understand how mutualistic microorganisms communicate with plants, how associated root developmental pathways are modulated, and how plant immune responses are successfully circumvented, many important questions remain. For example, little is currently known about more primitive modes of intercellular apoplastic colonization, whether for ectomycorrhizal fungi or for certain nitrogen-fixing symbioses. Neither do we know whether the CSSP has a key role in ectomycorrhizal associations, nor how host plants distinguish between structurally similar chitin-based “symbiotic” and “pathogenic” microbial signals. Answering these questions should contribute to our understanding of the underlying mechanisms that govern the relationships between plants and their entire microbiota. On a broader level, improved understanding of how environmental and genetic cues, together with plant metabolism, modulate microbial colonization will be crucial for the future exploitation of the microbiota for the benefit of sustainable plant growth. The major root symbioses established by land plants with soil microorganisms are arbuscular mycorrhizal, ectomycorrhizal, and nitrogen-fixing associations. Top left: Image of a mature arbuscule of the arbuscular mycorrhizal fungus Funneliformis mosseae in a plant root cell. Top right: Ectomycorrhizal rootlets of beech (Fagus sylvatica) in symbiosis with the ochre brittlegill fungus (Russula ochroleuca). Bottom left: Symbiotic root N-fixing legume nodules on a fava bean (Vicia faba) plant. Bottom right: Symbiotic N-fixing actinorhizal nodules on the root of an alder tree. CLOCKWISE FROM TOP LEFT: MARK BRUNDRETT; MARC BUÉE; CUSTOM LIFE SCIENCE IMAGES/ALAMY STOCK PHOTO; NIGEL CATTLIN/ALAMY STOCK PHOTO Within the plant microbiota, mutualistic fungal and bacterial symbionts are striking examples of microorganisms playing crucial roles in nutrient acquisition. They have coevolved with their hosts since initial plant adaptation to land. Despite the evolutionary distances that separate mycorrhizal and nitrogen-fixing symbioses, these associations share a number of highly conserved features, including specific plant symbiotic signaling pathways, root colonization strategies that circumvent plant immune responses, functional host-microbe interface formation, and the central role of phytohormones in symbiosis-associated root developmental pathways. We highlight recent and emerging areas of investigation relating to these evolutionarily conserved mechanisms, with an emphasis on the more ancestral mycorrhizal associations, and consider to what extent this knowledge can contribute to an understanding of plant-microbiota associations as a whole.

Specific impacts of beech and Norway spruce on the structure and diversity of the rhizosphere and soil microbial communities.

The impacts of plant species on the microbial communities and physico-chemical characteristics of soil are well documented for many herbs, grasses and legumes but much less so for tree species. Here, we investigate by rRNA and ITS amplicon sequencing the diversity of microorganisms from the three domains of life (Archaea, Bacteria and Eukaryota:Fungi) in soil samples taken from the forest experimental site of Breuil-Chenue (France). We discovered significant differences in the abundance, composition and structure of the microbial communities associated with two phylogenetically distant tree species of the same age, deciduous European beech (Fagus sylvatica) and coniferous Norway spruce (Picea abies Karst), planted in the same soil. Our results suggest a significant effect of tree species on soil microbiota though in different ways for each of the three microbial groups. Fungal and archaeal community structures and compositions are mainly determined according to tree species, whereas bacterial communities differ to a great degree between rhizosphere and bulk soils, regardless of the tree species. These results were confirmed by quantitative PCR, which revealed significant enrichment of specific bacterial genera, such as Burkholderia and Collimonas, known for their ability to weather minerals within the tree root vicinity.