Protein-RNA interface residue prediction using machine learning: an assessment of the state of the art

Background

Understanding the interaction among different species within a community and their responses to environmental changes is a central goal in ecology. However, defining the network structure in a microbial community is very challenging due to their extremely high diversity and as-yet uncultivated status. Although recent advance of metagenomic technologies, such as high throughout sequencing and functional gene arrays, provide revolutionary tools for analyzing microbial community structure, it is still difficult to examine network interactions in a microbial community based on high-throughput metagenomics data.

Results

Here, we describe a novel mathematical and bioinformatics framework to construct ecological association networks named molecular ecological networks (MENs) through Random Matrix Theory (RMT)-based methods. Compared to other network construction methods, this approach is remarkable in that the network is automatically defined and robust to noise, thus providing excellent solutions to several common issues associated with high-throughput metagenomics data. We applied it to determine the network structure of microbial communities subjected to long-term experimental warming based on pyrosequencing data of 16?S rRNA genes. We showed that the constructed MENs under both warming and unwarming conditions exhibited topological features of scale free, small world and modularity, which were consistent with previously described molecular ecological networks. Eigengene analysis indicated that the eigengenes represented the module profiles relatively well. In consistency with many other studies, several major environmental traits including temperature and soil pH were found to be important in determining network interactions in the microbial communities examined. To facilitate its application by the scientific community, all these methods and statistical tools have been integrated into a comprehensive Molecular Ecological Network Analysis Pipeline (MENAP), which is open-accessible now (http://ieg2.ou.edu/MENA).

Conclusions

The RMT-based molecular ecological network analysis provides powerful tools to elucidate network interactions in microbial communities and their responses to environmental changes, which are fundamentally important for research in microbial ecology and environmental microbiology.     

Investigation of organic anions in tree root exudates and rhizosphere microbial communities using in situ and destructive sampling techniques

Purpose

Understanding of the role of low molecular weight organic anions (OAs) in structuring rhizosphere microbial communities in situ is limited due to challenges associated with sampling. Improved techniques are needed for such studies.

Methods

This study used in situ and destructive sampling techniques and compared two exudate extraction methods [anion exchange membrane (AEM) capturing and water extraction] from rhizosphere and non-rhizosphere samples of genetically modified (GM) and control Pinus radiata D. Don trees grown in large-scale rhizotrons for ~10?months. Metabolically active soil microbial communities were analysed using rRNA-DGGE.

Results

Recovery of eight out of 12 anions was influenced by extraction methods, and in situ sampling using AEM was shown to be the most efficient method. Only minor differences were detected in OAs in root exudates collected from the GM and control trees. Significant differences in α-Proteobacterial and Pseudomonas communities were associated with the two tree lines in the topsoil at both sampling events. Additional differences in β-Proteobacterial and fungal communities between tree lines were detected in the rhizosphere using destructive sampling.

Conclusion

This study demonstrated that in situ sampling was superior to destructive sampling for the efficient collection of root exudates and analysis of associated rhizosphere microbial communities.     

环境DNA技术在地下生态学中的应用

地下生态过程是生态系统结构、功能和过程研究中最不确定的因素。由于技术和方法的限制,作为"黑箱"的地下生态系统已经成为限制生态学发展的瓶颈,也是未来生态学发展的主要方向。环境DNA技术,是指从土壤等环境样品中直接提取DNA片段,然后通过DNA测序技术来定性或定量化目标生物,以确定目标生物在生态系统中的分布及功能特征。环境DNA技术已成功用于地下生态过程的研究。目前,环境DNA技术在土壤微生物多样性及其功能方面的研究相对成熟,克服了土壤微生物研究中不能培养的问题,可以有效地分析土壤微生物的群落组成、多样性及空间分布,尤其是宏基因组学技术的发展,使得微生物生态功能方面的研究成为可能;而且,环境DNA技术已经在土壤动物生态学的研究中得到了初步应用,可快速分析土壤动物的多样性及其分布特征,更有效地鉴定出未知的或稀少的物种,鉴定土壤动物类群的幅度较宽;部分研究者通过提取分析土壤中DNA片段信息对生态系统植物多样性及植物分类进行了研究,其结果比传统的植物分类及物种多样性测定更精确,改变了以往对植物群落物种多样性模式的理解。同时,环境DNA技术克服传统根系研究方法中需要洗根、分根、只能测定单物种根系的局限,降低根系研究中细根区分的误差,并探索性地用于细根生物量的研究。主要综述了基于环境DNA技术的分子生物学方法在土壤微生物多样性及功能、土壤动物多样性、地下植物多样性及根系生态等地下生态过程研究中的应用进展。环境DNA技术对于以土壤微生物、土壤动物及地下植物根系为主体的地下生态学过程的研究具有革命性意义,并展现出良好的应用前景。可以预期,分子生物学技术与传统的生态学研究相结合将成为未来地下生态学研究的一个发展趋势。     

Composition and activity of rhizosphere microbial communities associated with healthy and diseased greenhouse tomatoes

Aims

The goal of this study was to investigate the structure and functional potential of microbial communities associated with healthy and diseased tomato rhizospheres.

Methods

Composition changes in the bacterial communities inhabiting the rhizospheric soil and roots of tomato plants were detected using 454 pyrosequencing. Microbial functional diversity was investigated with BIOLOG technology.

Results

There were significant shifts in the microbial composition of diseased samples compared with healthy samples, which had the highest bacterial diversity. The predominant phylum in both diseased and healthy samples was Proteobacteria, which accounted for 35.7–97.4 % of species. The class Gammaproteobacteria was more abundant in healthy than in diseased samples, while the Alphaproteobacteria and Betaproteobacteria were more abundant in diseased samples. The proportions of pathogenic Ralstonia solanacearum and Actinobacteria species were also elevated in diseased samples. The proportions of the various bacterial populations showed a similar trend both in rhizosphere soil and plant roots in diseased versus disease-free samples, indicating that pathogen infection altered the composition of bacterial communities in both plant and soil samples. In terms of microbial activity, functional diversity was suppressed in diseased soil samples. Soil enzyme activity, including urease, alkaline phosphatase and catalase activity, also declined.

Conclusions

This is the first report that provides evidence that R. solanacearum infection elicits shifts in the composition and functional potential of microbial communities in a continuous-cropping tomato operation.     

Growth response of crops to soil microbial communities from conventional monocropping and tree-based intercropping systems

Background and aims

Recent studies have shown that tree-based intercropping (TBI) systems support a more diverse soil microbial community compared to conventional agricultural systems. However, it is unclear whether differences in soil microbial diversity between these two agricultural systems have a functional effect on crop growth.

Methods

In this study, we used a series of greenhouse experiments to test whether crops respond differently to the total soil microbial community (Experiment 1) and to arbuscular mycorrhizal (AM) fungal communities alone (Experiment 2) from conventionally monocropped (CM) and TBI systems.

Results

The crops had a similar growth response to the total soil microbial communities from both cropping systems. However, when compared to sterilized controls, barley (Hordeum vulgare) and canola (Brassica napus) exhibited a negative growth response to the total soil microbial communities, while soybean (Glycine max) was unaffected. During the AM fungal establishment phase of the second experiment, ‘nurse’ plants had a strong positive growth response to AM fungal inoculation, and significantly higher biomass when inoculated with AM fungi from the CM system compared to the TBI system. Soybean was the only crop species to exhibit a significant positive growth response to AM fungal inoculation. Similar to the total soil microbial communities, AM fungi from the two cropping systems did not differ in their effect on crop growth.

Conclusion

Overall, AM fungi from both cropping systems had a positive effect on the growth of plants that formed a functional symbiosis. However, the results from these experiments suggest that negative effects of non-AM fungal microbes are stronger than the beneficial effects of AM fungi from these cropping systems.     

The root microbiota—a fingerprint in the soil?

Background

The root system of a plant is known to host a wide diversity of microbes that can be essential or detrimental to the plant. Microbial ecologists have long struggled to understand what factors structure the composition of these communities. An overlooked part of the microbial community succession in root systems has been the potential for individual variation among plants shaped by early colonisation events such as microbial exposure of the seed inside the parent plant and during dispersal.

Scope

In this review we outline life events of the plant that can affect the composition of its root microbiota and relate ecological theory of community assembly to the formation of the root microbiota.

Conclusion

All plants are exposed to environmental conditions and events throughout their lifetime that shape their phenotype. The microbial community associated with the plant is ultimately an extension of this phenotype. Therefore, only by following a plant from its origin inside the flower to senescence, can we fully understand how the associated microbial community was assembled and what determined its composition.     

Bacterial community in the rhizosphere of the cactus species Mammillaria carnea during dry and rainy seasons assessed by deep sequencing

Background and aims

The Tehuacán-Cuitcatlán reserve is an area of unique plant biodiversity mostly in the form of xerophytes, with exceptionally high numbers of rare and endemic species. This endemism results partly from the characteristics of the climate of this area, with two distinct seasons: rainy and dry seasons. Although rhizosphere communities must be critical in the function of this ecosystem, understanding the structure of these communities is currently limited. This is the first molecular study of the microbial diversity present in the rhizosphere of Mamillaria carnea.

Methods

Total DNA was obtained from soil and rhizosphere samples at three locations in the Tehuacán Cuicatlán Reserve, during dry and rainy seasons. Temperature gradient gel electrophoresisis (TGGE) fingerprinting, 16S rRNA gene libraries and pyrosequencing were used to investigate bacterial diversity in the rhizosphere of Mammillaria carnea and changes in the microbial community between seasons.

Results

Deep sequencing data reveal a higher level of biodiversity in the dry season. Statistical analyses based on these data indicates that the composition of the bacterial community differed between both seasons affecting to members of the phyla Acidobacteria, Cyanobacteria, Gemmatimonadetes, Plantomycetes, Actinobacteria and Firmicutes. In addition, the depth of sequencing performed (>24,000 reads) enables detection of changes in the relative abundance of lower bacterial taxa (novel bacterial phylotypes) indicative of the increase of specific bacterial populations due to the season.

Conclusions

This study states the basis of the bacterial diversity in the rhizosphere of cacti in semi-arid environments and it is a sequence-based demonstration of community shifts in different seasons.     

土壤动物肠道微生物多样性研究进展

随着分子生物学技术方法的快速发展,动物肠道微生物已成为医学、动物生理学与微生物生态学等研究领域热点。土壤动物种类繁多,分布广泛,其作为陆地生态系统重要组分,是驱动生态系统功能的关键因子。土壤动物体内的微生物由于与宿主长期共存,在与宿主协同进化中形成了丰富多样的群落结构,能够影响土壤动物本身的健康,进而介导土壤动物生态功能的实现。近些年,土壤动物肠道微生物工作方兴未艾,日渐得到重视。总结了四个部分内容：1)首先总结了土壤动物肠道微生物多样性领域的研究现状,该领域年发文量逐年增长,且近十年增长快速。土壤模式生物肠道微生物多样性研究较多且更为深入。土壤动物肠道微生物多样性组成与驱动机制、共存机制及群落构建的理论研究是该领域前沿;2)进而展示了土壤动物肠道微生物多样性组成和研究方法,土壤动物肠道菌群组成以变形菌门、厚壁菌门、放线菌门和拟杆菌门为主。早期工作基于传统分离培养,近年来新一代测序技术推动了该领域发展;3)接着关注了土壤动物肠道微生物的生态学功能,总体上体现在肠道微生物能帮助宿主分解食物基质、参与营养利用、影响寿命和繁殖及提高宿主免疫能力,且其能够影响土壤动物的气体排放及介导其对生态系...     

Metabolic and phylogenetic analysis of microbial communities during phytoremediation of soil contaminated with weathered hydrocarbons and heavy metals

In the current study, the microbial ecology of weathered hydrocarbon and heavy metal contaminated soil undergoing phytoremediation was studied. The relationship of functional diversity, measured as carbon source utilisation in Biolog plates and extracellular enzymatic activities, and genetic diversity of bacteria was evaluated. Denaturing gradient gel electrophoresis was used for community analyses at the species level. Bulk soil and rhizosphere soil from pine and poplar plantations were analysed separately to determine if the plant rhizosphere impacted hydrocarbon degradation. Prevailing microbial communities in the field site were both genetically and metabolically diverse. Furthermore, both tree rhizosphere and fertilisation affected the compositions of these communities and increased activities of extracellular aminopeptidases. In addition, the abundance of alkane hydroxylase and naphthalene dioxygenase genes in the communities was low, but the prevalence of these genes was increased by the addition of bioavailable hydrocarbons. Tree rhizosphere communities had greater hydrocarbon degradation potential than those of bulk soil. Hydrocarbon utilising communities were dominated generally by the species Ralstonia eutropha and bacteria belonging to the genus Burkholderia. Despite the presence of viable hydrocarbon-degrading microbiota, decomposition of hydrocarbons from weathered hydrocarbon contaminated soil over four years, regardless of the presence of vegetation, was low in unfertilised soil. Compost addition enhanced the removal of hydrocarbons.     

Roles of root border cells in plant defense and regulation of rhizosphere microbial populations by extracellular DNA ‘trapping’

Background

As roots penetrate soil, specialized cells called ‘border cells’ separate from root caps and contribute a large proportion of exudates forming the rhizosphere. Their function has been unclear. Recent findings suggest that border cells act in a manner similar to that of white blood cells functioning in defense. Histone-linked extracellular DNA (exDNA) and proteins operate as ‘neutrophil extracellular traps’ to attract and immobilize animal pathogens. DNase treatment reverses trapping and impairs defense, and mutation of pathogen DNase results in loss of virulence.

Scope

Histones are among a group of proteins secreted from living border cells. This observation led to the discovery that exDNA also functions in defense of root caps. Experiments revealed that exDNA is synthesized and exported into the surrounding mucilage which attracts, traps and immobilizes pathogens in a host-microbe specific manner. When this plant exDNA is degraded, the normal resistance of the root cap to infection is abolished.

Conclusions

Research to define how exDNA may operate in plant immunity is needed. In the meantime, the specificity and stability of exDNA and its association with distinct microbial species may provide an important new tool to monitor when, where, and how soil microbial populations become established as rhizosphere communities.     

Indirect effects of polycyclic aromatic hydrocarbon contamination on microbial communities in legume and grass rhizospheres

Background and aims

Biodegradation of polycyclic aromatic hydrocarbons (PAHs) is accelerated in the presence of plants, due to the stimulation of rhizosphere microbes by plant exudates (nonspecific enhancement). However, plants may also recruit specific microbial groups in response to PAH stress (specific enhancement). In this study, plant effects on the development of rhizosphere microbial communities in heterogeneously contaminated soils were assessed for three grasses (ryegrass, red fescue and Yorkshire fog) and four legumes (white clover, chickpea, subterranean clover and red lentil).

Methods

Plants were cultivated using a split-root model with their roots divided between two independent pots containing either uncontaminated soil or PAH-contaminated soil (pyrene or phenanthrene). Microbial community development in the two halves of the rhizosphere was assessed by T-RFLP (bacterial and fungal community) or DGGE (bacterial community), and by 16S rRNA gene tag-pyrosequencing.

Results

In legume rhizospheres, the microbial community structure in the uncontaminated part of the split-root model was significantly influenced by the presence of PAH-contamination in the other part of the root system (indirect effect), but this effect was not seen for grasses. In the contaminated rhizospheres, Verrucomicrobia and Actinobacteria showed increased populations, and there was a dramatic increase in Denitratisoma numbers, suggesting that this genus may be important in rhizoremediation processes.

Conclusion

Our results show that Trifolium and other legumes respond to PAH-contamination stress in a systemic manner, to influence the microbial diversity in their rhizospheres.     

Light exposure mediates circadian rhythms of rhizosphere microbial communities

Microbial community circadian rhythms have a broad influence on host health and even though light-induced environmental fluctuations could regulate microbial communities, the contribution of light to the circadian rhythms of rhizosphere microbial communities has received little attention. To address this gap, we monitored diel changes in the microbial communities in rice (Oryza sativa L.) rhizosphere soil under light–dark and constant dark regimes, identifying microbes with circadian rhythms caused by light exposure and microbial circadian clocks, respectively. While rhizosphere microbial communities displayed circadian rhythms under light–dark and constant dark regimes, taxa possessing circadian rhythms under the two conditions were dissimilar. Light exposure concealed microbial circadian clocks as a regulatory driver, leading to fewer ecological niches in light versus dark communities. These findings disentangle regulation mechanisms for circadian rhythms in the rice rhizosphere microbial communities and highlight the role of light-induced regulation of rhizosphere microbial communities.Subject terms: Microbial ecology, Community ecology     

Rice biomass production and carbon cycling in 13CO2 pulse-labeled microcosms with different soils under submerged conditions

Aims

Rice fields are an important source for the greenhouse gas methane. Plants play an essential role in carbon supply for soil microbiota, but the influence of the microbial community on carbon cycling is not well understood.

Methods

Microcosms were prepared using sand-vermiculite amended with different soils and sediments, and planted with rice. The microcosms at different growth stages were pulse-labeled with ¹³CO₂ followed by tracing ¹³C in plant, soil and atmospheric carbon pools and quantifying the abundance of methanogenic archaea in rhizosphere soil.

Results

Overall,?>85 % of the freshly assimilated carbon was allocated in aboveground plant biomass, approximately 10 % was translocated into the roots and?4, but emission of ¹³C-labeled CH₄ started immediately and ¹³C enrichment revealed that plant-derived carbon was an important source for methanogenesis. The results further demonstrated that carbon assimilation and translocation processes, microbial abundance and gas emission were not only affected by the plant growth stage, but also by the content and type of soil in which the rice plants grew.

Conclusions

The study illustrates the close ties between plant physiology, soil properties and microbial communities for carbon turnover and ecosystem functioning.     

Comparison of different methods for determining lignin concentration and quality in herbaceous and woody plant residues

Background

The development and dispersal of seeds as well as their transition to seedlings represent perhaps the most critical stages of a plant’s life cycle. The endophytic and epiphytic microbial interactions that take place in, on, and around seeds during these stages of the plant’s life cycle may have profound impacts on plant ecology, health, and productivity. While our understanding of the seed microbiota has lagged far behind that of the rhizosphere and phyllosphere, many advances are now being made.

Scope

This review explores the microbial associations with seeds through various stages of the plant life cycle, beginning with the earliest stages of seed development on the parent plant and continuing through the development and establishment of seedlings in soil. This review represents a broad synthesis of the ecological and agricultural literature focused on seed-microbe interactions as a means of better understanding how these interactions may ultimately influence plant ecology, health, and productivity in both natural and agricultural systems. Our current understanding of seed-microbe associations will be discussed, with an emphasis on recent findings that specifically highlight the emerging contemporary understanding of how seed-microbe associations may ultimately impact plant health and productivity.

Conclusions

The diversity and dynamics of seed microbiomes represent the culmination of complex interactions with microbes throughout the plant life cycle. The richness and dynamics of seed microbiomes is revealing exciting new opportunities for research into plant-microbe interactions. Often neglected in plant microbiome studies, the renaissance of inquiry into seed microbiomes is offering exciting new insights into how the diversity and dynamics of the seed microbiome with plant and soil microbiomes as well as the microbiomes of dispersers and pollinators. It is clear that the interactions taking place in and around seeds indeed have significant impacts on plant health and productivity in both agricultural and natural ecosystems.

     

The influence of litter quality on the relationship between vegetation and below-ground compartments: a Procrustean approach

Aims

We used a Procrustean superimposition approach associated with regression analysis to test hypotheses regarding the relationship between plant communities and distinct below-ground compartments—soil chemistry (SC) and soil microbial activity (SMA). Additionally, we evaluated litter chemical quality as an interface between the above and below-ground compartments.

Methods

Plant community, and soil chemical and biochemical data from three post-mining degraded sites under reclamation and from one nearby forest site in the Brazilian Amazon Basin were analyzed.

Results

All studied sites presented distinct plant community, litter quality, SC and SMA. Plant community consistently affected the below-ground variation in both SC and SMA compartments. The influence of litter quality was greater in the plant community versus SMA relationship than in the plant community versus SC. Nevertheless, the SC affected significantly the SMA, but without influence of litter quality.

Conclusions

Differently from previous studies, our findings suggest that plant community and soil chemistry can affect the soil microbial activity independently. Specifically for our study area, these results point to a rupture of the ‘in nested’ structure of the causal relationship between changes in vegetation, changes in the chemical litter quality, changes in the SC and the response of SMA.     

Depth-Dependent Differences in Community Structure of the Human Colonic Microbiota in Health

Objective

The aims of this study were to develop techniques for spatial microbial assessment in humans and to establish colonic luminal and mucosal spatial ecology, encompassing longitudinal and cross-sectional axes.

Design

A microbiological protected specimen brush was used in conjunction with a biopsy forceps to sample the colon in nine healthy volunteers undergoing colonoscopy. Terminal Restriction Fragment Length Polymorphism analysis was used to determine the major variables in the spatial organization of the colonic microbiota.

Results

Protected Specimen Brush sampling retrieved region-specific, uncontaminated samples that were enriched for bacterial DNA and depleted in human DNA when compared to biopsy samples. Terminal Restriction Fragment Length Polymorphism analysis revealed a segmentation of bacterial communities between the luminal brush and biopsy-associated ecological niches with little variability across the longitudinal axis of the colon and reduced diversity in brush samples.

Conclusion

These results support the concept of a microbiota with little longitudinal variability but with some degree of segregation between luminal and mucosal communities.     

Predicting Ecological Roles in the Rhizosphere Using Metabolome and Transportome Modeling

The ability to obtain complete genome sequences from bacteria in environmental samples, such as soil samples from the rhizosphere, has highlighted the microbial diversity and complexity of environmental communities. However, new algorithms to analyze genome sequence information in the context of community structure are needed to enhance our understanding of the specific ecological roles of these organisms in soil environments. We present a machine learning approach using sequenced Pseudomonad genomes coupled with outputs of metabolic and transportomic computational models for identifying the most predictive molecular mechanisms indicative of a Pseudomonad’s ecological role in the rhizosphere: a biofilm, biocontrol agent, promoter of plant growth, or plant pathogen. Computational predictions of ecological niche were highly accurate overall with models trained on transportomic model output being the most accurate (Leave One Out Validation F-scores between 0.82 and 0.89). The strongest predictive molecular mechanism features for rhizosphere ecological niche overlap with many previously reported analyses of Pseudomonad interactions in the rhizosphere, suggesting that this approach successfully informs a system-scale level understanding of how Pseudomonads sense and interact with their environments. The observation that an organism’s transportome is highly predictive of its ecological niche is a novel discovery and may have implications in our understanding microbial ecology. The framework developed here can be generalized to the analysis of any bacteria across a wide range of environments and ecological niches making this approach a powerful tool for providing insights into functional predictions from bacterial genomic data.     

Effects of above-ground plant species composition and diversity on the diversity of soil-borne microorganisms

A coupling of above-ground plant diversity and below-ground microbial diversity has been implied in studies dedicated to assessing the role of macrophyte diversity on the stability, resilience, and functioning of ecosystems. Indeed, above-ground plant communities have long been assumed to drive below-ground microbial diversity, but to date very little is known as to how plant species composition and diversity influence the community composition of micro-organisms in the soil. We examined this relationship in fields subjected to different above-ground biodiversity treatments and in field experiments designed to examine the influence of plant species on soil-borne microbial communities. Culture-independent strategies were applied to examine the role of wild or native plant species composition on bacterial diversity and community structure in bulk soil and in the rhizosphere. In comparing the influence of Cynoglossum officinale (hound's tongue) and Cirsium vulgare (spear thistle) on soil-borne bacterial communities, detectable differences in microbial community structure were confined to the rhizosphere. The colonisation of the rhizosphere of both plants was highly reproducible, and maintained throughout the growing season. In a separate experiment, effects of plant diversity on bacterial community profiles were also only observed for the rhizosphere. Rhizosphere soil from experimental plots with lower macrophyte diversity showed lower diversity, and bacterial diversity was generally lower in the rhizosphere than in bulk soil. These results demonstrate that the level of coupling between above-ground macrophyte communities and below-ground microbial communities is related to the tightness of the interactions involved. Although plant species composition and community structure appear to have little discernible effect on microbial communities inhabiting bulk soil, clear and reproducible changes in microbial community structure and diversity are observed in the rhizosphere. This revised version was published online in August 2006 with corrections to the Cover Date.