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The microbiota populating the plant–soil continuum defines an untapped resource for sustainable crop production. The host plant is a driver for the taxonomic composition and function of these microbial communities. In this review, we illustrate how the host genetic determinants of the microbiota have been shaped by plant domestication and crop diversification. We discuss how the heritable component of microbiota recruitment may represent, at least partially, a selection for microbial functions underpinning the growth, development, and health of their host plants and how the magnitude of this heritability is influenced by the environment. We illustrate how host–microbiota interactions can be treated as an external quantitative trait and review recent studies associating crop genetics with microbiota-based quantitative traits. We also explore the results of reductionist approaches, including synthetic microbial communities, to establish causal relationships between microbiota and plant phenotypes. Lastly, we propose strategies to integrate microbiota manipulation into crop selection programs. Although a detailed understanding of when and how heritability for microbiota composition can be deployed for breeding purposes is still lacking, we argue that advances in crop genomics are likely to accelerate wider applications of plant–microbiota interactions in agriculture. Expected final online publication date for the Annual Review of Phytopathology, Volume 61 is September 2023. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

Plant root-associated bacteria can confer protection against pathogen infection. By contrast, the beneficial effects of root endophytic fungi and their synergistic interactions with bacteria remain poorly defined. We demonstrate that the combined action of a fungal root endophyte from a widespread taxon with core bacterial microbiota members provides synergistic protection against an aggressive soil-borne pathogen inArabidopsis thalianaand barley. We additionally reveal early inter-kingdom growth promotion benefits which are host and microbiota composition dependent. Using RNA-sequencing, we show that these beneficial activities are not associated with extensive host transcriptional reprogramming but rather with the modulation of expression of microbial effectors and carbohydrate-active enzymes.

Plant exudates are one of the main drivers for the proliferation of the rhizosphere microbiota. We developed a protocol to quantify primary metabolites in exudates of the global crop barley. We used 2- and 3-week-old barley plants grown in sterilised sand and their root exudates were collected for 6 hours in ddH2O water to avoid degradation/transformation of the compounds. These samples were used to determine total carbon and total nitrogen by Dumas. Finally, we profiled primary metabolites in collected exudates by gas chromatography/mass spectrometry (GC-MS).

Background The microbiota thriving in the rhizosphere, the thin layer of soil surrounding plant roots, plays a critical role in plant’s adaptation to the environment. Domestication and breeding selection have progressively differentiated the microbiota of modern crops from the ones of their wild ancestors. However, the impact of eco-geographical constraints faced by domesticated plants and crop wild relatives on recruitment and maintenance of the rhizosphere microbiota remains to be fully elucidated. Methods We grew twenty wild barley ( Hordeum vulgare ssp. spontaneum ) genotypes representing five distinct ecogeographic areas in the Israeli region, one of the sites of barley domestication, alongside four ’Elite’ varieties ( H. vulgare ssp. vulgare ) in a previously characterised agricultural soil under greenhouse conditions. At early stem elongation, rhizosphere samples were collected, and stem and root dry weight measured. In parallel, we generated high-resolution 16S rRNA gene profiles of the rhizosphere and unplanted soil samples. Ecological indices and multivariate statistical analyses allowed us to identify ‘host signatures’ for the composition of the rhizosphere microbiota. Finally, we capitalised on single nucleotide polymorphisms (SNPs) of the barley genome to investigate the relationships between microbiota diversity and host genetic diversity. Results Elite material outperformed the wild genotypes in aboveground biomass while, almost invariably, wild genotypes allocated more resources to belowground growth. These differential growth responses were associated with a differential microbial recruitment in the rhizosphere. The selective enrichment of individual bacterial members of microbiota mirrored the distinct ecogeographical constraints faced by the wild and domesticated plants. Unexpectedly, Elite varieties exerted a stronger genotype effect on the rhizosphere microbiota when compared with wild barley genotypes adapted to desert environments and this effect had a bias for Actinobacteria . Finally, in wild barley genotypes, we discovered a limited, but significant, correlation between microbiota diversity and host genomic diversity. Conclusions Our results revealed a footprint of the host’s adaptation to the environment on the assembly of the bacteria thriving at the root-soil interface. This recruitment cue layered atop of the distinct evolutionary trajectories of wild and domesticated plants and, at least in part, is encoded by the barley genome. This knowledge will be critical to further dissect microbiota contribution to plant’s adaptation to the environment and to devise strategies for climate-smart agriculture.

A culture collection of 41 bacteria isolated from the rhizosphere of cultivated barley ( Hordeum vulgare subsp. vulgare ) is available at the Division of Plant Sciences, University of Dundee (UK). The data include information on plant growth-promoting genes implicated in nitrogen fixation, HCN channels, phosphate solubilization, and linked whole-genome sequences.

Otiorhynchus sulcatus (Fabricius) (Coleoptera: Curculionidae), commonly known as black vine weevil or simply vine weevil, is an important pest of soft fruit and ornamental crops. This species is endemic to temperate areas of Europe but has spread to many other areas over the last century, including North America and Australasia. The ability of vine weevils to adapt to such different environments is difficult to reconcile with the parthenogenetic reproduction strategy, which is likely to underpin a low genetic diversity. It is therefore tempting to hypothesize that weevil adaptation to different environments is mediated, at least partly, by the microbial communities inhabiting these insects. As a first step towards testing this hypothesis we characterized the composition of the bacterial microbiota in weevils from populations feeding on strawberry plants across four geographically separate locations in the UK. We performed 16S rRNA gene Illumina amplicon sequencing, generating 2 882 853 high‐quality reads. Ecological indices, namely Chao1 and Shannon, revealed that the populations used for this study harboured a low diversity and an uneven bacterial microbiota. Furthermore, β‐diversity analysis failed to identify a clear association between microbiota composition and location. Notably, a single operational taxonomic unit phylogenetically related to Candidatus Nardonella accounted for 81% of the total sequencing reads for all tested insects. Our results indicate that vine weevil bacterial microbiota resembles that of other insects as it has low diversity and it is dominated by few taxa. A prediction of this observation is that location per se may not be a determinant of the microbiota inhabiting weevil populations. Rather, other or additional selective pressures, such as the plant species used as a food source, ultimately shape the weevil bacterial microbiota. Our results will serve as a reference framework to investigate other or additional hypotheses aimed at elucidating vine weevil adaptation to its environment.

Plant root-associated bacteria can confer protection against pathogen infection. By contrast, the beneficial effects of root endophytic fungi and their synergistic interactions with bacteria remain poorly defined. We demonstrate that the combined action of a fungal root endophyte from a widespread taxon with core bacterial microbiota members provides synergistic protection against an aggressive soil-borne pathogen in Arabidopsis thaliana and barley. We additionally reveal early inter-kingdom growth promotion benefits which are host and microbiota composition dependent. Using RNA-sequencing, we show that these beneficial activities are not associated with extensive host transcriptional reprogramming but rather with the modulation of expression of microbial effectors and carbohydrate-active enzymes.

Plant root-associated bacteria can confer protection against pathogen infection. By contrast, the beneficial effects of root endophytic fungi and their synergistic interactions with bacteria remain poorly defined. We demonstrate that the combined action of a fungal root endophyte from a widespread taxon with core bacterial microbiota members provides synergistic protection against an aggressive soil-borne pathogen inArabidopsis thalianaand barley. We additionally show early inter-kingdom growth promotion benefits which are host and microbiota composition dependent.HighlightsThe root endophytic fungusSerendipita vermiferacan functionally replace core bacterial microbiota members in mitigating pathogen infection and disease symptoms.S. vermiferaadditionally stabilizes and potentiates the protective activities of root-associated bacteria and mitigates the negative effects of a non-native bacterial community inA. thaliana.Inter-kingdom synergistic beneficial effects do not require extensive host transcriptional reprogramming nor high levels ofS. vermiferacolonisation.Inter-kingdom protective benefits are largely independent of the host while synergism leading to early inter-kingdom growth promotion is driven by host species and microbiota composition.

We generated a bacterial collection from the rhizosphere of cultivated barley (Hordeum vulgare L. ssp. vulgare) to assess taxonomic distribution of culturable members of the barley microbiota and their plant growth-promoting potential. From this we retrieved strains belonging to the dominant phyla of the plant microbiota— Actinobacteria, Bacteroidetes, Firmicutes and Proteobacteria—and gathered evidence they code for functional genes implicated in nitrogen fixation, hydrogen cyanide channels and phosphate solubilisation. Here we present an initial comparative genomic analysis of the collection revealing that plant growth-promoting potential of the culturable barley bacterial microbiota appears to have a relatively broad phylogenetic base while retaining some strain-specificity.

Microbial communities proliferating at the root-soil interface, collectively referred to as the rhizosphere microbiota, represent an untapped beneficial resource for plant growth, development and health. Integral to a rational manipulation of the microbiota for sustainable agriculture is the identification of the molecular determinants of these communities. In plants, biosynthesis of allelochemicals is centre stage in defining inter-organismal relationships in the environment. Intriguingly, this process has been moulded by domestication and breeding selection. The indole-alkaloid gramine, whose occurrence in barley (Hordeum vulgare L.) is widespread among wild genotypes but has been counter selected in several modern varieties, is a paradigmatic example of this phenomenon. This prompted us to investigate how exogenous applications of gramine impacted on the rhizosphere microbiota of two, gramine-free, elite barley varieties grown in a reference agricultural soil. High throughput 16S rRNA gene amplicon sequencing revealed that applications of gramine interfere with the proliferation of a subset of soil microbes with a relatively broad phylogenetic assignment. Strikingly, growth of these bacteria appeared to be rescued by barley plants in a genotype- and dosage-independent manner. In parallel, we discovered that host recruitment cues can interfere with the impact of gramine application in a host genotype-dependent manner. Interestingly, this latter effect displayed a bias for members of the phyla Proteobacteria. These initial observations indicate that gramine can act as a determinant of the prokaryotic communities inhabiting the root-soil interface.

Background Phosphorus (P) availability is the main nutritional factor that limits crops yields in tropical soils due to edaphic processes that lead to P immobilization after mineral fertilization. Considering the potential of the rhizosphere microbiome to transform insoluble P into forms readily available for plant uptake, in this study is proposed that plants with contrasting P uptake efficiency, growing under depleted amounts of P are able to shape distinct bacterial communities in the rhizosphere enriching taxa specialized in P mobilization. Methods We selected two common bean genotypes contrasting in P efficiency uptake and grew them in a soil with a gradient of two different sources of P, triple superphosphate (TSP) or rock phosphate Bayovar (RPB). The rhizosphere bacterial community was assessed by 16S rRNA amplicon sequencing. Data analyses focused in describing the structure of the bacterial communities, identification of OTUs differentially enriched in different treatments, functional metagenomic prediction and cooccurrence network. Results P sources and levels resulted in different rhizosphere bacterial community structure. A high number of differentially enriched OTUs were observed under P depleted conditions in the P-inefficient genotype, mainly belonging to Actinobacteria phylum. The P-inefficient genotype did not show significant differences in the rhizosphere bacterial community assembly growing in different P sources. Predicted metagenome profiles showed the enrichment of bacterial functions involved in P mobilization, in the rhizosphere of the P inefficient genotype cultivated in P depleted conditions. The network analysis revealed that in the rhizosphere of the P-inefficient genotype under P depleted conditions the bacterial community has a higher number of nodes and edges, higher average degree and clustering coeficient when compared to the treatment with optimal P level. Conclusion Our data showed that the uptake of exogenous input resulted in the assembly of a P-competent microbiome in the P-inefficient genotype compared to the efficient one, supporting the hypothesis that the selective pressure for the P uptake engages P-inefficient genotypes in symbiotic relationships with the soil microbiome. These results will pave the way for future experimentation aiming at explore the contribution of this P-competent microbiome to plant growth and development in a range of soil type.

The microbiota thriving at the root-soil interface plays a crucial role in supporting plant growth, development and health. The interactions between plant and soil microbes can be traced back to the initial plant's colonisation of dry lands. Understanding the evolutionary drivers of these interactions will be key to re-wire them for the benefit of mankind. Here we critically assess recent insights into the evolutionary history of plant-microbiota interactions in natural and agricultural ecosystems. We identify distinctive features, as well as commonalities, of these two distinct scenarios and areas requiring further research efforts. Finally, we propose strategies that combining advances in molecular microbiology and crop genomics will be key towards a predictable manipulation of plant-microbiota interactions for sustainable crop production.

The projected increase in world population and the need to reduce the reliance on non-renewable inputs, such as synthetic agrochemicals, are challenging the current vision of agriculture. In particular, to achieve a fair and sustainable global food security, disruptive changes in crop production are unavoidable. A promising strategy proposes to exploit the metabolic capabilities of soil microbial communities, i.e., the microbiome, to conjugate stable yield with reduced impact on the agroecosystem. In this chapter, we introduce the microbiome populating the root-soil interface from an evolutionary perspective. Next, we discuss the molecular bases of plant-microbe interactions in soil and how these interactions impact plant growth, development and health. We illustrate how plant-probiotic members of the microbiome can be isolated from soil and further characterized for their biological activities, a key pre-requisite for translational applications. In addition, we focus on paradigmatic examples of soil microbes turned into inoculants for agriculture, their fate on soil, their impact on the native microbiome and the beneficial effects exerted on crop production.

Plants host distinct microbial communities on and inside their tissues designated the plant microbiota. Microbial community profiling enabled the description of the phylogenetic structure of the plant microbiota to an unprecedented depth, whereas functional insights are largely derived from experiments using individual microorganisms. The binary interplay between isolated members of the plant microbiota and host plants ranges from mutualistic to commensalistic and pathogenic relationships. However, how entire microbial communities capable of executing both growth-promoting and growth-compromising activities interfere with plant fitness remains largely unknown. Ultimately, unravelling the net result of microbial activities encoded in the extended plant genome—the plant microbiome—will be key to understanding and exploiting the full yield potential of a crop plant. In this perspective, we summarize first achievements of plant-microbiome research, we discuss future research directions, and we provide ideas for the translation of basic science to application to capitalize on the plant microbiome at work.

Phragmites australis and Typha latifolia are two macrophytes commonly present in natural and artificial wetlands. Roots of these plants engage in interactions with a broad range of microorganisms, collectively referred to as the microbiota. The microbiota contributes to the natural process of phytodepuration, whereby pollutants are removed from contaminated water bodies through plants. The outermost layer of the root corpus, the rhizoplane, is a hot-spot for these interactions where microorganisms establish specialized aggregates designated biofilm. Earlier studies suggest that biofilm-forming members of the microbiota play a crucial role in the process of phytodepuration. However, the composition and recruitment cue of the Phragmites, and Typha microbiota remain poorly understood. We therefore decided to investigate the composition and functional capacities of the bacterial microbiota thriving at the P. australis and T. latifolia root–soil interface. By using 16S rRNA gene Illumina MiSeq sequencing approach we demonstrated that, despite a different composition of the initial basin inoculum, the microbiota associated with the rhizosphere and rhizoplane of P. australis and T. latifolia tends to converge toward a common taxonomic composition dominated by members of the phyla Actinobacteria, Firmicutes, Proteobacteria, and Planctomycetes. This indicates the existence of a selecting process acting at the root–soil interface of these aquatic plants reminiscent of the one observed for land plants. The magnitude of this selection process is maximum at the level of the rhizoplane, where we identified different bacteria enriched in and discriminating between rhizoplane and rhizosphere fractions in a species-dependent and -independent way. This led us to hypothesize that the structural diversification of the rhizoplane community underpins specific metabolic capabilities of the microbiota. We tested this hypothesis by complementing the sequencing survey with a biochemical approach and scanning electron microscopy demonstrating that rhizoplane-enriched bacteria have a bias for biofilm-forming members. Together, our data will be critical to facilitate the rational exploitation of plant–microbiota interactions for phytodepuration.

peer-reviewed Gaining a greater understanding of the plant microbiota and its interactions with its host plant heralds a new era of scientific discovery in agriculture. Different agricultural management practices influence soil microbial populations by changing a soil’s physical, chemical and biological properties. However, the impact of these practices on the microbiota associated with economically important crops such as oilseed rape, are still understudied. In this work we investigated the impact of two contrasting crop establishment practices, conventional (plow based) and conservation (strip–tillage) systems, on the microbiota inhabiting different plant microhabitats, namely rhizosphere, root and shoot, of winter oilseed rape under Irish agronomic conditions. Illumina 16S rRNA gene sequence profiling showed that the plant associated microhabitats (root and shoot), are dominated by members of the bacterial phyla Proteobacteria, Actinobacteria and Bacteroidetes. The root and shoot associated bacterial communities displayed markedly distinct profiles as a result of tillage practices. We observed a very limited ‘rhizosphere effect’ in the root zone of WOSR, i.e., there was little or no increase in bacterial community richness and abundance in the WOSR rhizosphere compared to the bulk soil. The two tillage systems investigated did not appear to lead to any major long term differences on the bulk soil or rhizosphere bacterial communities. Our data suggests that the WOSR root and shoot microbiota can be impacted by management practices and is an important mechanism that could allow us to understand how plants respond to different management practices and environments. This work was supported through the Teagasc Walsh Fellowship Scheme which funded RR. The analysis of the sequencing data was supported by Royal Society of Edinburgh/Scottish Government Personal Research Fellowship co-funded by Marie Curie Actions awarded to DB.

This work was supported by a Royal Society of Edinburgh/Scottish Government Personal Research Fellowship co-funded by Marie Curie Actions awarded to DB. SR-A is supported by a BBSRC iCASE studentship awarded to DB (BB/M016811/1) and partnered by the James Hutton Limited (Invergowrie, United Kingdom). RAT is supported by a Scottish Food Security Alliance-Crops studentship, provided by the University of Dundee, the University of Aberdeen, and the James Hutton Institute. James Hutton researchers receive financial support from the Rural and Environment Science and Analytical Service Division of the Scottish Government., The rhizosphere, the thin layer of soil surrounding and influenced by plant roots, defines a distinct and selective microbial habitat compared to unplanted soil. The microbial communities inhabiting the rhizosphere, the rhizosphere microbiota, engage in interactions with their host plants which span from parasitism to mutualism. Therefore, the rhizosphere microbiota emerges as one of the determinants of yield potential in crops. Studies conducted with different plant species have unequivocally pointed to the host plant as a driver of the microbiota thriving at the root–soil interface. Thus far, the host genetic traits shaping the rhizosphere microbiota are not completely understood. As root hairs play a critical role in resource exchanges between plants and the rhizosphere, we hypothesized that they can act as a determinant of the microbiota thriving at the root–soil interface. To test this hypothesis, we took advantage of barley (Hordeum vulgare) mutant lines contrasting for their root hair characteristics. Plants were grown in two agricultural soils, differentiating in their organic matter contents, under controlled environmental conditions. At early stem elongation rhizosphere specimens were collected and subjected to high-resolution 16S rRNA gene profiling. Our data revealed that the barley rhizosphere microbiota is largely dominated by members of the phyla Bacteroidetes and Proteobacteria, regardless of the soil type and the root hair characteristics of the host plant. Conversely, ecological indices calculated using operational taxonomic units (OTUs) presence, abundance, and phylogeny revealed a significant impact of root hair mutations on the composition of the rhizosphere microbiota. In particular, our data indicate that mutant plants host a reduced-complexity community compared to wild-type genotypes and unplanted soil controls. Congruently, the host genotype explained up to 18% of the variation in ecological distances computed for the rhizosphere samples. Importantly, this effect is manifested in a soil-dependent manner. A closer inspection of the sequencing profiles revealed that the root hair-dependent diversification of the microbiota is supported by a taxonomically narrow group of bacteria, with a bias for members of the orders Actinomycetales, Burkholderiales, Rhizobiales, Sphingomonadales, and Xanthomonadales. Taken together, our results indicate that the presence and function of root hairs are a determinant of the bacterial community thriving in the rhizosphere and their perturbations can markedly impact on the recruitment of individual members of the microbiota., Royal Society of Edinburgh/Scottish Government Personal Research Fellowship

A major challenge facing agriculture in the 21st century is the need to increase the productivity of cultivated land while reducing the environmentally harmful consequences of mineral fertilization. The microorganisms thriving in association and interacting with plant roots, the plant microbiota, represent a potential resource of plant probiotic function, capable of conjugating crop productivity with sustainable management in agroecosystems. However, a limited knowledge of the organismal interactions occurring at the root-soil interface is currently hampering the development and use of beneficial plant-microbiota interactions in agriculture. Therefore, a comprehensive understanding of the recruitment cues of the plant microbiota and the molecular basis of nutrient turnover in the rhizosphere will be required to move toward efficient and sustainable crop nutrition. In this chapter, we will discuss recent insights into plant-microbiota interactions at the root-soil interface, illustrate the processes driving mineral dynamics in soil, and propose experimental avenues to further integrate the metabolic potential of the plant microbiota into crop management and breeding strategies for sustainable agricultural production.

The plant root defines the interface between a multicellular eukaryote and soil, one of the richest microbial ecosystems on Earth. Notably, soil bacteria are able to multiply inside roots as benign endophytes and modulate plant growth and development, with implications ranging from enhanced crop productivity to phytoremediation. Endophytic colonization represents an apparent paradox of plant innate immunity because plant cells can detect an array of microbe-associated molecular patterns (also known as MAMPs) to initiate immune responses to terminate microbial multiplication. Several studies attempted to describe the structure of bacterial root endophytes; however, different sampling protocols and low-resolution profiling methods make it difficult to infer general principles. Here we describe methodology to characterize and compare soil- and root-inhabiting bacterial communities, which reveals not only a function for metabolically active plant cells but also for inert cell-wall features in the selection of soil bacteria for host colonization. We show that the roots of Arabidopsis thaliana, grown in different natural soils under controlled environmental conditions, are preferentially colonized by Proteobacteria, Bacteroidetes and Actinobacteria, and each bacterial phylum is represented by a dominating class or family. Soil type defines the composition of root-inhabiting bacterial communities and host genotype determines their ribotype profiles to a limited extent. The identification of soil-type-specific members within the root-inhabiting assemblies supports our conclusion that these represent soil-derived root endophytes. Surprisingly, plant cell-wall features of other tested plant species seem to provide a sufficient cue for the assembly of approximately 40% of the Arabidopsis bacterial root-inhabiting microbiota, with a bias for Betaproteobacteria. Thus, this root sub-community may not be Arabidopsis-specific but saprophytic bacteria that would naturally be found on any plant root or plant debris in the tested soils. By contrast, colonization of Arabidopsis roots by members of the Actinobacteria depends on other cues from metabolically active host cells.

SUMMARY Barley ( Hordeum vulgare L.) leaf stripe is caused by the seed-borne fungus Pyrenophora graminea . We investigated microscopically and molecularly the reaction of barley embryos to leaf stripe inoculation. In the resistant genotype NIL3876- Rdg2a , fungal growth ceased at the scutellar node of the embryo, while in the susceptible near-isogenic line (NIL) Mirco- rdg2a fungal growth continued past the scutellar node and into the embryo. Pathogenchallenged embryos of resistant and susceptible NILs showed different levels of UV autofluorescence and toluidine blue staining, indicating differential accumulation of phenolic compounds. Suppression subtractive hybridization and cDNA amplified fragment-length polymorphism (AFLP) analyses of embryos identified P. graminea -induced and P. graminea -repressed barley genes. In addition, cDNA-AFLP analysis identified six pathogenicityassociated fungal genes expressed during barley infection but at low to undetectable levels during growth on artificial media. Microarrays representing the entire set of differentially expressed cDNA-AFLP fragments and 100 barley homologues of previously described defence-related genes were used to study gene expression changes at 7 and 14 days after inoculation in the resistant and susceptible NILs. A total of 171 significantly modulated barley genes were identified and assigned to four groups based on timing and genotype dependence of expression. Analysis of the changes in gene expression during the barley resistance response to leaf stripe suggests that the Rdg2a -mediated response includes cell-wall reinforcement, signal transduction, generation of reactive oxygen species, cell protection, jasmonate signalling and expression of plant effector genes. The identification of genes showing leaf stripe inoculation or resistance-dependent expression sets the stage for further dissection of the resistance response of barley embryo cells to leaf stripe.

Summary The microbial communities inhabiting the root interior of healthy plants, as well as the rhizosphere, which consists of soil particles firmly attached to roots, engage in symbiotic associations with their host. To investigate the structural and functional diversification among these communities, we employed a combination of 16S rRNA gene profiling and shotgun metagenome analysis of the microbiota associated with wild and domesticated accessions of barley (Hordeum vulgare). Bacterial families Comamonadaceae, Flavobacteriaceae, and Rhizobiaceae dominate the barley root-enriched microbiota. Host genotype has a small, but significant, effect on the diversity of root-associated bacterial communities, possibly representing a footprint of barley domestication. Traits related to pathogenesis, secretion, phage interactions, and nutrient mobilization are enriched in the barley root-associated microbiota. Strikingly, protein families assigned to these same traits showed evidence of positive selection. Our results indicate that the combined action of microbe-microbe and host-microbe interactions drives microbiota differentiation at the root-soil interface., Graphical Abstract, Highlights • A small number of bacterial families dominate the root-enriched barley microbiota • The host genotype determines the profile of a subset of community members • Functions relevant for host interactions are enriched in root-associated taxa • Genes mediating host, bacteria, and phage interactions show signs of positive selection, Microbial communities inhabiting the root interior and surrounding soil contribute to plant growth. Bulgarelli et al. examine the microbiota that populates the roots of barley (Hordeum vulgare) and present evidence that integrated actions of microbe-microbe and host-microbe interactions drive root microbiota establishment through physiological processes occurring at the root-soil interface.

Leaf stripe disease on barley is caused by the seed-transmitted hemi-biotrophic fungus Pyrenophora graminea. Race-specific resistance to leaf stripe is controlled by two known Rdg (resistance to Drechslera graminea) genes: the H. spontaneum-derived Rdg1a, mapped to chromosome 2HL and Rdg2a, identified in H. vulgare, mapped on chromosome 7HS. Both resistance genes have been extensively used in classical breeding. The positional cloning and molecular characterization of the Rdg2a locus is described here. BAC and cosmid libraries, respectively, derived from barley cvs. Morex (susceptible to leaf stripe) and Thibaut (the donor of the Rdg2a allele) were used for physical mapping of Rdg2a. At the Rdg2a locus, three sequence-related coiled-coil, nucleotide-binding site and leucine-rich repeat (CC-NB-LRR) encoding genes were identified. Sequence comparisons suggested that paralogs of this resistance locus evolved through recent gene duplication and were subjected to frequent sequence exchange. Transformation of the leaf stripe susceptible cv. Golden Promise with two Rdg2a candidates identified a member of the CC-NB-LRR gene family that conferred resistance against the Dg2 leaf stripe isolate, towards which the Rdg2a gene is effective. Histological analysis demonstrated that Rdg2a-mediated leaf stripe resistance prevents pathogen colonisation in the embryos without any detectable hypersensitive cell death response, indicating an unusual resistance mechanism for a CC-NB-LRR protein.

BACKGROUND: Leaf stripe disease on barley (Hordeum vulgare) is caused by the seed-transmitted hemi-biotrophic fungus Pyrenophora graminea. Race-specific resistance to leaf stripe is controlled by two known Rdg (Resistance to Drechslera graminea) genes: the H. spontaneum-derived Rdg1a and Rdg2a, identified in H. vulgare. The aim of the present work was to isolate the Rdg2a leaf stripe resistance gene, to characterize the Rdg2a locus organization and evolution and to elucidate the histological bases of Rdg2a-based leaf stripe resistance. PRINCIPAL FINDINGS: We describe here the positional cloning and functional characterization of the leaf stripe resistance gene Rdg2a. At the Rdg2a locus, three sequence-related coiled-coil, nucleotide-binding site, and leucine-rich repeat (CC-NB-LRR) encoding genes were identified. Sequence comparisons suggested that paralogs of this resistance locus evolved through recent gene duplication, and were subjected to frequent sequence exchange. Transformation of the leaf stripe susceptible cv. Golden Promise with two Rdg2a-candidates under the control of their native 5' regulatory sequences identified a member of the CC-NB-LRR gene family that conferred resistance against the Dg2 leaf stripe isolate, against which the Rdg2a-gene is effective. Histological analysis demonstrated that Rdg2a-mediated leaf stripe resistance involves autofluorescing cells and prevents pathogen colonization in the embryos without any detectable hypersensitive cell death response, supporting a cell wall reinforcement-based resistance mechanism. CONCLUSIONS: This work reports about the cloning of a resistance gene effective against a seed borne disease. We observed that Rdg2a was subjected to diversifying selection which is consistent with a model in which the R gene co-evolves with a pathogen effector(s) gene. We propose that inducible responses giving rise to physical and chemical barriers to infection in the cell walls and intercellular spaces of the barley embryo tissues represent mechanisms by which the CC-NB-LRR-encoding Rdg2a gene mediates resistance to leaf stripe in the absence of hypersensitive cell death.

Genetic mapping of major genes and quantitative traits loci (QTLs) for many important agricultural traits is increasing the integration of biotechnology with the conventional breeding process. Exploitation of the information derived from the map position of traits with agronomical importance and of the linked molecular markers, can be achieved through marker assisted selection (MAS) of the traits during the breeding process. However, empirical applications of this procedure have shown that the success of MAS depends upon several factors, including the genetic base of the trait, the degree of the association between the molecular marker and the target gene, the number of individuals that can be analyzed and the genetic background in which the target gene has to be transferred. MAS for simply inherited traits is gaining increasing importance in breeding programs, allowing an acceleration of the breeding process. Traits related to disease resistance to pathogens and to the quality of some crop products are offering some important examples of a possible routinary application of MAS. For more complex traits, like yield and abiotic stress tolerance, a number of constraints have determined severe limitations on an efficient utilization of MAS in plant breeding, even if there are a few successful applications in improving quantitative traits. Recent advances in genotyping technologies together with comparative and functional genomic approaches are providing useful tools for the selection of genotypes with superior agronomical performancies.

The dominant gene Rdg2a of barley conferring resistance to the hemi-biotrophic seed-borne pathogen Pyrenophora graminea is located in the distal region of chromosome arm 1 (7H)S. As the first step towards isolating the gene, a high-resolution genetic map of the region was constructed using an F(2) population of 1,400 plants (Thibaut Rdg2axMirco). The map included six classes of resistance gene analogues (RGAs) tightly associated with Rdg2a. Rdg2a was delimited to a genetic interval of 0.14 cM between the RGAs ssCH4 and MWG851. Additional markers were generated using the sequence from the corresponding region on rice chromosome 6, allowing delimitation of the Rdg2a syntenic interval in rice to a 115 kbp stretch of sequence. Analysis of the rice sequence failed to reveal any genes with similarity to characterized resistance genes. Therefore, either the rice-barley synteny is disrupted in this region, or Rdg2a encodes a novel type of resistance protein.

Made available in DSpace on 2020-12-10T17:31:35Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-01 GENBACCA project (Regione Emilia Romagna, POR-FESR 2014/2020 GENBACCA Initiative) Royal Society of Edinburgh/Scottish Government Personal Research Fellowship Marie Actions Biotechnology and Biological Sciences Research Council (BBSRC) iCASE studentship Horizon 2020 Framework Programme Innovation Action 'CIRCLES' (European Commission) The microbial communities thriving at the root-soil interface have the potential to improve plant growth and sustainable crop production. Yet, how agricultural practices, such as the application of either mineral or organic nitrogen fertilizers, impact on the composition and functions of these communities remains to be fully elucidated. By deploying a two-pronged 16S rRNA gene sequencing and predictive metagenomics approach, we demonstrated that the bacterial microbiota of field-grown tomato (Solarium tycopersicum) plants is the product of a selective process that progressively differentiates between rhizosphere and root microhabitats. This process initiates as early as plants are in a nursery stage and it is then more marked at late developmental stages, in particular at harvest. This selection acts on both the bacterial relative abundances and phylogenetic assignments, with a bias for the enrichment members of the phylum Actinobacteria in the root compartment. Digestate-based and mineral-based nitrogen fertilizers trigger a distinct bacterial enrichment in both rhizosphere and root microhabitats. This compositional diversification mirrors a predicted functional diversification of the root-inhabiting communities, manifested predominantly by the differential enrichment of genes associated to ABC transporters and the two-component system. Together, our data suggest that the microbiota thriving at the tomato root soil interface is modulated by and in responses to the type of nitrogen fertilizer applied to the field. Univ Modena & Reggio Emilia, Ctr BIOGEST SITEIA, Dept Life Sci, Reggio Emilia, Italy Univ Dundee, Sch Life Sci, Plant Sci, Dundee, Scotland Sao Paulo State Univ, Fac Agr & Vet Sci, Jaboticabal, Brazil Sao Paulo State Univ, Fac Agr & Vet Sci, Jaboticabal, Brazil Biotechnology and Biological Sciences Research Council (BBSRC) iCASE studentship: BB/M016811/1 Horizon 2020 Framework Programme Innovation Action 'CIRCLES' (European Commission): 818290

Plants host distinct bacterial communities on and inside various plant organs, of which those associated with roots and the leaf surface are best characterized. The phylogenetic composition of these communities is defined by relatively few bacterial phyla, including Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria. A synthesis of available data suggests a two-step selection process by which the bacterial microbiota of roots is differentiated from the surrounding soil biome. Rhizodeposition appears to fuel an initial substrate-driven community shift in the rhizosphere, which converges with host genotype–dependent fine-tuning of microbiota profiles in the selection of root endophyte assemblages. Substrate-driven selection also underlies the establishment of phyllosphere communities but takes place solely at the immediate leaf surface. Both the leaf and root microbiota contain bacteria that provide indirect pathogen protection, but root microbiota members appear to serve additional host functions through the acquisition of nutrients from soil for plant growth. Thus, the plant microbiota emerges as a fundamental trait that includes mutualism enabled through diverse biochemical mechanisms, as revealed by studies on plant growth–promoting and plant health–promoting bacteria.

The current population growth will demand a higher productive agriculture to full the food requirement. To supply this need and preserve the environment, many resources are applied to promote sustainable agriculture. Phosphorus depletion is the main factor that limits crops yields in tropical soils, where the pH and clay content rapid fixate this nutrient. Plant breeders aim to solve this issue by changing the plant requirements for phosphorus and adapting them to low P availability. However, with these approaches the demand for phosphorus fertilizers will continue and so the depletion of the natural deposits. In this study is proposed that plants with contrasting phosphorus uptake efficiency, i.e. P-efficient and P-inefficient, recruits distinct rhizosphere microbiome specialized in phosphorus mobilization. This hypothesis was tested growing plants in a gradient of two sources of P, triple superphosphate or rock phosphate Bayovar. Thebean rhizosphere microbiome was assessed with culture dependent and independent approaches, enzymatic assays, predictive metagenomics and networks analysis. A differential enrichment of several OTUs in the rhizosphere of the P-inefficient common bean genotype, and the enrichment of bacterial chemotaxis functions and functions involved in phosphorus mobilization suggest that this genotype has superior communication with the rhizosphere microbiome and is highly dependent on it for phosphorus mobilization. As a proof of concept, the P-efficientefficient genotype was sown in soil previously cultivated with P-inefficientinefficient genotype. The results showed that P-efficientefficient genotype positively responded to the modified rhizosphere in early stages, that is, the microbiome selected and enriched by the P-inefficient genotype improved the P uptake in the genotype cultivated afterwards in the same soil. Taken collectively, these results suggest that plants partly rely on the rhizosphere microbiome for P uptake and that the exploration of these interactions during plant breeding would allow the selection of even more efficient genotypes, leading to a sustainable agriculture by exploring soil residual P. O atual aumento populacional irá demandar uma maior produção agrícola para completar a necessidade de alimento. Para suprir essa necessidade e preservar o meio ambiente, muitos recursos serão aplicados para promover a agricultura sustentável. A depleção de fósforo é um dos principais fatores que limita a produção agrícola em solos tropicais, onde o pH e o conteúdo de argila fixam rapidamente esse nutriente. Os melhoristas de plantas visam solucionar esse problema alterando a necessidade de fósforo das plantas e adaptando-as as baixas disponibilidade de fósforo. No entanto, com essas estratégias a demanda por fertilizantes fosfatados irá continuar assim como a exploração das reservas naturais de fósforo. Nesse estudo foi proposto que as plantas contrastantes em relação a eficiência na absorção de fósforo, i.e. P-eficiente e P-ineficiente, recrutariam um microbioma rizosférico distinto em relação a mobilização de fósforo. Essa hipótese foi testada cultivando plantas em um gradiente usando duas fontes distintas de P, triplo fosfato ou fosfato de rocha Bayovar. O microbioma da rizosfera de feijão foi então avaliado por técnicas dependentes e independentes de cultivo, análise enzimática, predição metagenômica e análises de network. Um enriquecimento diferencial de várias OTUs observado na rizosfera do genótipo de feijão P-ineficiente, e o enriquecimento de funções de quimiotaxia bacteriana e envolvidas na mobilização de fósforo sugerem que esse genótipo tem uma maior comunicação com o microbioma rizosférico e é altamente dependente deste para a mobilização de fósforo. Como prova de conceito, o genótipo P-eficiente foi plantado em solo previamente cultivadocom o genótipo P-ineficiente. Os resultados mostraram que o genótipo P-eficiente responde positivamente à rizosfera modificada nos estádios iniciais de crescimento, ou seja, o microbioma selecionado e enriquecido pelo genótipo P-ineficiente melhorou a absorção de fósforo no genótipo cultivado posteriormente no mesmo solo. Coletivamente, esses resultados sugerem que as plantas dependem parcialmente do microbioma da rizosfera para a absorção de P e que a exploraçãodestas interações durante o melhoramento vegetal permitiria a seleção de genótipos muito mais eficientes, conduzindo à uma agricultura sustentável explorando o fósforo residual do solo.