Plant-associated Microbiomes Research Articles

Core microbiomes for sustainable agroecosystems.

In an era of ecosystem degradation and climate change, maximizing microbial functions in agroecosystems has become a prerequisite for the future of global agriculture. However, managing species-rich communities of plant-associated microbiomes remains a major challenge. Here, we propose interdisciplinary research strategies to optimize microbiome functions in agroecosystems. Informatics now allows us to identify members and characteristics of 'core microbiomes', which may be deployed to organize otherwise uncontrollable dynamics of resident microbiomes. Integration of microfluidics, robotics and machine learning provides novel ways to capitalize on core microbiomes for increasing resource-efficiency and stress-resistance of agroecosystems.

Plants for Sustainable Improvement of Indoor Air Quality.

Indoor pollution poses a serious threat to human health. Plants represent a sustainable but underexploited solution to enhance indoor air quality. However, the current selection of plants suitable for indoors fails to consider the physiological processes and mechanisms involved in phytoremediation. Therefore, the capacity of plants to remove indoor air pollutants through stomatal uptake (absorption) and non-stomatal deposition (adsorption) remains largely unknown. Moreover, the effects of the indoor plant-associated microbiome still need to be fully analyzed. Here, we discuss how a combination of the enhanced phytoremediation capacity of plants together with cutting-edge air-cleaning and smart sensor technologies can improve indoor life while reducing energy consumption.

Drought delays development of the sorghum root microbiome and enriches for monoderm bacteria

Drought stress is a major obstacle to crop productivity, and the severity and frequency of drought are expected to increase in the coming century. Certain root-associated bacteria have been shown to mitigate the negative effects of drought stress on plant growth, and manipulation of the crop microbiome is an emerging strategy for overcoming drought stress in agricultural systems, yet the effect of drought on the development of the root microbiome is poorly understood. Through 16S rRNA amplicon and metatranscriptome sequencing, as well as root metabolomics, we demonstrate that drought delays the development of the early sorghum root microbiome and causes increased abundance and activity of monoderm bacteria, which lack an outer cell membrane and contain thick cell walls. Our data suggest that altered plant metabolism and increased activity of bacterial ATP-binding cassette (ABC) transporter genes are correlated with these shifts in community composition. Finally, inoculation experiments with monoderm isolates indicate that increased colonization of the root during drought can positively impact plant growth. Collectively, these results demonstrate the role that drought plays in restructuring the root microbiome and highlight the importance of temporal sampling when studying plant-associated microbiomes.

The effects of host species and sexual dimorphism differ among root, leaf and flower microbiomes of wild strawberries in situ

Plant-associated microbiomes profoundly influence host interactions with below- and aboveground environments. Characterizing plant-associated microbiomes in experimental settings have revealed important drivers of microbiota assemblies within host species. However, it remains unclear how important these individual drivers (e.g., organ type, host species, host sexual phenotype) are in structuring the patterns of plant–microbiota association in the wild. Using 16s rRNA sequencing, we characterized root, leaf and flower microbiomes in three closely related, sexually polymorphic Fragaria species, in the broadly sympatric portion of their native ranges in Oregon, USA. Taking into account the potential influence of broad-scale abiotic environments, we found that organ type explained the largest variation of compositional and phylogenetic α- and β-diversity of bacterial communities in these wild populations, and its overall effect exceeded that of host species and host sex. Yet, the influence of host species increased from root to leaf to flower microbiomes. We detected strong sexual dimorphism in flower and leaf microbiomes, especially in host species with the most complete separation of sexes. Our results provide the first demonstration of enhanced influence of host species and sexual dimorphism from root to flower microbiomes, which may be applicable to many other plants in the wild.

Community analysis of Persian oak fungal microbiome under dust storm conditions

Abstract Dust storms have major effects on terrestrial ecosystems through the long-distance transport and deposition of particulate matter. It is unclear how dust deposition affects plant-associated microbiomes in downwind ecosystems. Here we show that dust deposition may negatively influence the isolation, richness, and diversity of endophytic fungal communities of Persian oak. We used culture-based methods paired with Sanger sequencing to examine these effects on fungal assemblages isolated from leaf, branch and deposited dust. Increased amounts of dust deposition led to decreased endophytic fungal diversity in plant tissues but increased fungal diversity in deposited dust layers on leaves. Dust deposition decreased the abundance of a dominant endophyte in branches with promising biocontrol properties. Endophytic fungal communities found in leaves were more similar to fungal assemblages of deposited dust in comparison to branch endophytes. Our results suggest that dust storms may have ecosystem-wide effects by altering the fungal microbiomes of forest-forming trees.

Identifying the plant-associated microbiome across aquatic and terrestrial environments: the effects of amplification method on taxa discovery.

Plants in terrestrial and aquatic environments contain a diverse microbiome. Yet, the chloroplast and mitochondria organelles of the plant eukaryotic cell originate from free-living cyanobacteria and Rickettsiales. This represents a challenge for sequencing the plant microbiome with universal primers, as ~99% of 16S rRNA sequences may consist of chloroplast and mitochondrial sequences. Peptide nucleic acid clamps offer a potential solution by blocking amplification of host-associated sequences. We assessed the efficacy of chloroplast and mitochondria-blocking clamps against a range of microbial taxa from soil, freshwater and marine environments. While we found that the mitochondrial blocking clamps appear to be a robust method for assessing animal-associated microbiota, Proteobacterial 16S rRNA binds to the chloroplast-blocking clamp, resulting in a strong sequencing bias against this group. We attribute this bias to a conserved 14-bp sequence in the Proteobacteria that matches the 17-bp chloroplast-blocking clamp sequence. By scanning the Greengenes database, we provide a reference list of nearly 1500 taxa that contain this 14-bp sequence, including 48 families such as the Rhodobacteraceae, Phyllobacteriaceae, Rhizobiaceae, Kiloniellaceae and Caulobacteraceae. To determine where these taxa are found in nature, we mapped this taxa reference list against the Earth Microbiome Project database. These taxa are abundant in a variety of environments, particularly aquatic and semiaquatic freshwater and marine habitats. To facilitate informed decisions on effective use of organelle-blocking clamps, we provide a searchable database of microbial taxa in the Greengenes and Silva databases matching various n-mer oligonucleotides of each PNA sequence.

Discovery of Polyesterases from Moss-Associated Microorganisms.

In this study, we describe the discovery and analysis of new enzymes from microbial communities associated with plants (moss). The recovered enzymes show the ability to hydrolyze not only common esterase substrates but also the synthetic polyester poly(butylene adipate-co-butylene terephthalate), which is a common material employed in biodegradable plastics. The widespread use of such synthetic polyesters in industry and society requires the development of new sustainable technological solutions for their recycling. The discovered enzymes have the potential to be used as catalysts for selective recovery of valuable building blocks from this material.

Rhizosphere Sampling Protocols for Microbiome (16S/18S/ITS rRNA) Library Preparation and Enrichment for the Isolation of Drought Tolerance-Promoting Microbes.

Natural plant microbiomes are abundant and have a remarkably robust composition, both as epiphytes on the plant surface and as endophytes within plant tissues. Microbes in the former "habitat" face limited nutrients and harsh environmental conditions, while those in the latter likely lead a more sheltered existence. The most populous and diverse of these microbiomes are associated with the zone around the plant roots, commonly referred to as the rhizosphere. A majority of recent studies characterize these plant-associated microbiomes by community profiling of bacteria and fungi, using amplicon-based marker genes and next-generation sequencing (NGS). Here, we collate a group of protocols that incorporate current best practices and optimized methodologies for sampling, handling of samples, and rRNA library preparation for variable regions of V5-V6 and V9 of the bacterial 16S ribosomal RNA (rRNA) gene, and the ITS2 region joining the 5.8S and 28S regions of the fungal rRNA gene. Samples collected for such culture-independent analyses can also be used for the actual isolation of microbes of interest, perhaps even those identified by the libraries described above. One group of microbes that holds promise for mediating plant stress incurred by drought are bacteria that are capable of reducing or eliminating the plant's perception of the stress through degradation of the gaseous plant hormone ethylene, which is abundantly produced in response to drought stimuli. This is accomplished by some types of soil bacteria that can produce the enzyme 1-aminocyclopropane-1-carboxylate (ACC) deaminase, which is the immediate precursor to ethylene. Here we provide a high-throughput protocol for screening of ACC deaminase-producing bacteria for the applied purpose of mitigating the impact of plant drought stress.

The nutrient preference of plants influences their rhizosphere microbiome

Many studies in recent decades have shown the signature effect of the host plant in determining the plant-associated microbiome in the soil. However, the important question as to the factors contributing to the selective enrichment of microorganisms in the plant rhizosphere has not been fully addressed. In this study, the role of the nutrient preferences of two plant species, tomato and cucumber, in variations in the soil microbiome were investigated using a five-season continuous pot experiment. The results of MiSeq sequencing showed that these two plants assembled specific bacterial and fungal communities in their rhizospheres, and the soil nutrient status resulting from the plant nutrient preference was identified as a key driver in the development of a plant-specific microbiome.

High diversity of enzymes in plant-associated microbiomes

High diversity of enzymes in plant-associated microbiomes

Towards an Enhanced Understanding of Plant-Microbiome Interactions to Improve Phytoremediation: Engineering the Metaorganism.

Phytoremediation is a promising technology to clean-up contaminated soils based on the synergistic actions of plants and microorganisms. However, to become a widely accepted, and predictable remediation alternative, a deeper understanding of the plant–microbe interactions is needed. A number of studies link the success of phytoremediation to the plant-associated microbiome functioning, though whether the microbiome can exist in alternative, functional states for soil remediation, is incompletely understood. Moreover, current approaches that target the plant host, and environment separately to improve phytoremediation, potentially overlook microbial functions and properties that are part of the multiscale complexity of the plant-environment wherein biodegradation takes place. In contrast, in situ studies of phytoremediation research at the metaorganism level (host and microbiome together) are lacking. Here, we discuss a competition-driven model, based on recent evidence from the metagenomics level, and hypotheses generated by microbial community ecology, to explain the establishment of a catabolic rhizosphere microbiome in a contaminated soil. There is evidence to ground that if the host provides the right level and mix of resources (exudates) over which the microbes can compete, then a competitive catabolic and plant-growth promoting (PGP) microbiome can be selected for as long as it provides a competitive superiority in the niche. The competition-driven model indicates four strategies to interfere with the microbiome. Specifically, the rhizosphere microbiome community can be shifted using treatments that alter the host, resources, environment, and that take advantage of prioritization in inoculation. Our model and suggestions, considering the metaorganism in its natural context, would allow to gain further knowledge on the plant–microbial functions, and facilitate translation to more effective, and predictable phytotechnologies.

Transplanting Soil Microbiomes Leads to Lasting Effects on Willow Growth, but not on the Rhizosphere Microbiome.

Plants interact closely with microbes, which are partly responsible for plant growth, health, and adaptation to stressful environments. Engineering the plant-associated microbiome could improve plant survival and performance in stressful environments such as contaminated soils. Here, willow cuttings were planted into highly petroleum-contaminated soils that had been gamma-irradiated and subjected to one of four treatments: inoculation with rhizosphere soil from a willow that grew well (LA) or sub-optimally (SM) in highly contaminated soils or with bulk soil in which the planted willow had died (DE) or no inoculation (CO). Samples were taken from the starting inoculum, at the beginning of the experiment (T0) and after 100 days of growth (TF). Short hypervariable regions of archaeal/bacterial 16S rRNA genes and the fungal ITS region were amplified from soil DNA extracts and sequenced on the Illumina MiSeq. Willow growth was monitored throughout the experiment, and plant biomass was measured at TF. CO willows were significantly smaller throughout the experiment, while DE willows were the largest at TF. Microbiomes of different treatments were divergent at T0, but for most samples, had converged on highly similar communities by TF. Willow biomass was more strongly linked to overall microbial community structure at T0 than to microbial community structure at TF, and the relative abundance of many genera at T0 was significantly correlated to final willow root and shoot biomass. Although microbial communities had mostly converged at TF, lasting differences in willow growth were observed, probably linked to differences in T0 microbial communities.

Plant-associated microbiomes in arid lands: diversity, ecology and biotechnological potential

Aridification is a worldwide serious threat directly affecting agriculture and crop production. In arid and desert areas, it has been found that microbial diversity is huge, built of microorganisms able to cope with the environmental harsh conditions by developing adaptation strategies. Plants growing in arid lands or regions facing prolonged abiotic stresses such as water limitation and salt accumulation have also developed specific physiological and molecular stress responses allowing them to thrive under normally unfavorable conditions. Under such extreme selection pressures, special root-associated bacterial assemblages, endowed with capabilities of plant growth promotion (PGP) and extremophile traits, are selected by the plants. In this review, we provide a general overview on the microbial diversity in arid lands and deserts versus specific microbial assemblages associated with plants. The ecological drivers that shape this diversity, how plant-associated microbiomes are selected, and their biotechnological potential are discussed. Selection and recruitment of the plant associated bacterial assemblages is mediated by the combination of the bio-pedo-agroclimatic conditions and the plant species or varieties. Diversity and functional redundancy of these associated PGPR makes them very active in supporting plant improvement, health and resistance to drought, salt and related stresses. Implementing proper biotechnological applications of the arid and desert-adapted PGPR constitute the challenge to be raised.

Editorial: Plant responses to bacterial quorum sensing molecules.

Editorial: Plant responses to bacterial quorum sensing molecules.

Agroforestry leads to shifts within the gammaproteobacterial microbiome of banana plants cultivated in Central America.

Bananas (Musa spp.) belong to the most important global food commodities, and their cultivation represents the world's largest monoculture. Although the plant-associated microbiome has substantial influence on plant growth and health, there is a lack of knowledge of the banana microbiome and its influencing factors. We studied the impact of (i) biogeography, and (ii) agroforestry on the banana-associated gammaproteobacterial microbiome analyzing plants grown in smallholder farms in Nicaragua and Costa Rica. Profiles of 16S rRNA genes revealed high abundances of Pseudomonadales, Enterobacteriales, Xanthomonadales, and Legionellales. An extraordinary high diversity of the gammaproteobacterial microbiota was observed within the endophytic microenvironments (endorhiza and pseudostem), which was similar in both countries. Enterobacteria were identified as dominant group of above-ground plant parts (pseudostem and leaves). Neither biogeography nor agroforestry showed a statistically significant impact on the gammaproteobacterial banana microbiome in general. However, indicator species for each microenvironment and country, as well as for plants grown in Coffea intercropping systems with and without agri-silvicultural production of different Fabaceae trees (Inga spp. in Nicaragua and Erythrina poeppigiana in Costa Rica) could be identified. For example, banana plants grown in agroforestry systems were characterized by an increase of potential plant-beneficial bacteria, like Pseudomonas and Stenotrophomonas, and on the other side by a decrease of Erwinia. Hence, this study could show that as a result of legume-based agroforestry the indigenous banana-associated gammaproteobacterial community noticeably shifted.

Future challenges and perspectives for applying microbial biotechnology in sustainable agriculture based on a better understanding of plant-microbiome interactions

An intensive agricultural production is necessary to satisfy food requirements for the growing world population. However, its realization is associated with the mass consumption of non-renewable natural resources and with the emission of greenhouse gases causing climate changes. The research challenge is to meet sustainable environmental and economical issues without compromising yields. In this context, exploiting the agro-ecosystem services ofsoil microbial communities appears as a promising effective approach. This chapter reviews the research efforts aimed atimproving a sustainable and healthy agricultural production through the appropriate management of soil microorganisms.First, the plant-associated microbiome is briefly described. Then, the current research technologies for formulation and application of inocula based on specific beneficial plant-associated microbesare summarized. Finally, the perspectives and opportunities to manage naturally existing microbial populations, including those non-culturable, are analyzed. This analysis concerns: (i) a description of the already available, culture-independent, molecular techniques addressed at increasing our understanding of root-microbiome interactions; (ii) how to improve the ability of soil microbes for alleviating the negative impacts of stress factors on crop productivity; and (iii) whether plants can structure their root-associated microbial communities and, leading on from this, whether the rhizosphere can be engineered (biased) to encourage beneficial organisms, while prevent presence of pathogens.

The plant microbiome and its importance for plant and human health.

To study plant-associated microorganisms has a long history that reaches back to Lorenz Hiltner's definition of the rhizosphere in 1904 (Hartmann et al., 2008). Today, we know that microorganisms colonizing plant surfaces and inner tissues play an eminent role in shaping of our planet—from our natural vegetation to intense agricultural production systems up to human health. Plant-associated microorganisms have to be considered as key drivers for plant health, productivity, community composition, and ecosystem functioning. For this e-book “The plant microbiome and its importance for plant and human health” we collected 18 articles, including reviews, original, and opinion articles that highlight the current knowledge regarding plant microbiomes, their specificity, diversity, and function as well as all aspects studying the management of plant microbiomes to improve plant performance and health. The contribution of the single articles of this research topic to these questions is discussed in detail in the mini-review and 1st chapter of the book by Berg et al. (2014a). Overall the presented articles confirm that the plant-associated microbiome has greatly expanded the metabolic repertoire of plants and often increase resource uptake and provide novel nutritional and defense pathways. Thus, the plant microbiome has a direct impact on plant functional traits, such as leaf longevity, specific leaf area, leaf nutrient levels, and shoot/root ratio. By providing novel nutritional and defense pathways and by modifying biochemical pathways, the plant associated microbiome can enhance or decrease species coexistence and consequently influence not only a single plant but complete ecosystems. Thus, future breeding strategies may take the importance of plant-microbe interactions more into account than in the past, to obtain plants that generate high yields and are more tolerate to the constraints of global change. Studies related to raw-eaten vegetables are a special show case in this e-book. Here the plant-associated microbiome does not only influence plant performance but strongly contributes to human health. As those microbes are also part of our diet they can either improve human health (Blaser et al., 2013) or cause heavy outbreaks of infectious diseases by transferring possible pathogens (Van Overbeek et al., 2014). Interestingly, the gathered manuscripts indicate that microbiomes of different environments are not isolated but show interplay. For example, the microbiome of vegetables, humans as well as build environment such as hospitals seems to be well-connected (Berg et al., 2014b). Thus, maintaining microbial diversity in the different environments is an important issue to avoid pathogen outbreaks, which can be often explained by microbial imbalances and poorness (Van Elsas et al., 2012), confirming basic theories of ecology that a loss of native species enhances the probability of invasive species to colonize new environments. Therefore, to maintain and support microbial diversity is of interest to stabilize ecosystems and their resilience toward biotic and abiotic stressors. Biotechnological solutions like probiotics, prebiotics, and synbiotics for plants as well as humans can provide support for the indigenous microbiome (De Vrese and Schrezenmeir, 2008). The most significant recent advances in plant microbiology involve interdisciplinary approaches that link different methodological approaches including omics-technologies. Due to the new methods available and interdisciplinary research cooperation we have the chance to solve many problems of a changing world, but also to address basic hypotheses and questions of microbial ecology and host microbe interactions. Integrating epigenetics in multi-omics techniques opens existing opportunities for new discoveries (Chen et al., 2014). Therefore, we think this comprehensive e-book especially the many reviews can contribute to hold the current knowledge in our hand. This is a very exciting but also challenging time for all researchers in this field. Major advances will come rapidly!

Beneficial effects of plant-associated microbes on indoor microbiomes and human health?

Just like humans, plants have recently been recognized as meta-organisms, possessing a distinct microbiome and revealing close symbiotic relationships with their associated microorganisms (Berg et al., 2013; Mendes et al., 2013). Each plant harbor specific species to a certain degree but also cosmopolitan and ubiquitous microbial strains; the majority of them fulfill important host as well as ecosystem functions (rev. in Berg and Smalla, 2009). In addition to the microbe-rich rhizosphere, which has been studied extensively, the phyllosphere is of special interest for the study of indoor microbiomes due to its large and exposed surface area and its remarkable microbial diversity (Lindow and Leveau, 2002; Lindow and Brandl, 2003; Redford et al., 2010; Meyer and Leveau, 2012; Vorholt, 2012; Rastogi et al., 2013). In addition to the majority of beneficial and neutral inhabitants, all plant-associated microbiomes contain plant as well as human pathogens (Berg et al., 2005; Mendes et al., 2013). A broad spectrum of plant pathogens is well-known from disease outbreaks. Human pathogens belong mainly to the so called opportunistic or facultative human pathogens such as Burkholderia cepacia, Pseudomonas aeruginosa or Stenotrophomonas maltophilia, which cause diseases only in patients with predisposition or in hospital (Berg et al., 2005; Ryan et al., 2009). Microbiomes of humans and plants are currently intensively studied using the same methods and addressing similar scientific questions (Ramirez-Puebla et al., 2013). However, knowledge about the microbiomes' interaction, microbial dynamics and exchange in a certain biotope or even indoor environment is very much limited. Although the composition and function of plant microbiomes is well-studied, there is still little to no information regarding their overlap, interaction with -and impact on other microbiomes or the microbiome-harboring hosts. Information is available about the connection of soil and rhizosphere microbial diversity, which share a selective sub-set (Smalla et al., 2001). The root-soil interface is the selection site for plant-associated bacteria by root exudates, which acts as chemo-attractants as well as repellents to which bacteria respond (Badri and Vivanco, 2009). In addition, plant defense signaling play a role in this process (Doornbos et al., 2012). For the phyllosphere we know that there is only a part of residents, while a substantial part of bacteria is shared with the air microbiome (Lindow and Brandl, 2003). Based on these data, a strong interaction and exchange of rhizosphere and phyllosphere microbiomes with other microbiomes is obvious. However, this opinion paper focuses on the question, if there is also a connection from plant–to indoor microbiomes as well as an impact on human health.

The root microbiome influences scales from molecules to ecosystems: The unseen majority1

Plants are teeming with microbial organisms including those that colonize internal tissues as well as those that adhere to external surfaces. In the rhizosphere, the plant-associated microbiome is intricately involved in plant health and serves as a reservoir of additional genes that plants can access when needed. Microbiome regulation of plant trait expression affects plant performance, which in turn influences various ecosystem functions, such as primary productivity and soil health. Understanding these plant- and microbe-driven interactions requires a study of the nature and effects of the plant microbiome. Conceptualizing the microbiome requires a synthesis of microbial ecology, physiology, and bioinformatics, integrated with insight into host biology and ecology. Microbiome structure and function analyses are recognized as essential components to understand the genetic and functional capacity of the host (previously assigned solely to the host) and include vital aspects of metabolism and physiology. Here, as a special section, we present a set of papers that address the complex interactions between plants and root microbiomes in the rhizosphere. This unseen majority spans scales; with its microorganisms numerically dominant in terrestrial ecosystems, the root microbiome is also involved in plant genetics through integral roles in plant trait expression that can effect community composition and ecosystem functions, such as soil health.

The microbiome of medicinal plants: diversity and importance for plant growth, quality and health

Past medicinal plant research primarily focused on bioactive phytochemicals, however, the focus is currently shifting due to the recognition that a significant number of phytotherapeutic compounds are actually produced by associated microbes or through interaction with their host. Medicinal plants provide an enormous bioresource of potential use in modern medicine and agriculture, yet their microbiome is largely unknown. The objective of this review is (i) to introduce novel insights into the plant microbiome with a focus on medicinal plants, (ii) to provide details about plant- and microbe-derived ingredients of medicinal plants, and (iii) to discuss possibilities for plant growth promotion and plant protection for commercial cultivation of medicinal plants. In addition, we also present a case study performed both to analyse the microbiome of three medicinal plants (Matricaria chamomilla L., Calendula officinalis L., and Solanum distichum Schumach. and Thonn.) cultivated on organically managed Egyptian desert farm and to develop biological control strategies. The soil microbiome of the desert ecosystem was comprised of a high abundance of Gram-positive bacteria of prime importance for pathogen suppression under arid soil conditions. For all three plants, we observed a clearly plant-specific selection of the microbes as well as highly specific diazotrophic communities that overall identify plant species as important drivers in structural and functional diversity. Lastly, native Bacillus spec. div. strains were able to promote plant growth and elevate the plants’ flavonoid production. These results underline the numerous links between the plant-associated microbiome and the plant metabolome.