Marsh Rhizosphere Research Articles

Diversity at single nucleotide to pangenome scales among sulfur cycling bacteria in salt marshes.

Salt marshes are known for their significant carbon storage capacity, and sulfur cycling is closely linked with the ecosystem-scale carbon cycling in these ecosystems. Sulfate reducers are key for the decomposition of organic matter, and sulfur oxidizers remove toxic sulfide, supporting the productivity of marsh plants. To date, the complexity of coastal environments, heterogeneity of the rhizosphere, high microbial diversity, and uncultured majority hindered our understanding of the genomic diversity of sulfur-cycling microbes in salt marshes. Here, we use comparative genomics to overcome these challenges and provide an in-depth characterization of sulfur-cycling microbial diversity in salt marshes. We characterize communities across distinct sites and plant species and uncover extensive genomic diversity at the taxon level and specific genomic features present in MAGs affiliated with uncultivated sulfur-cycling lineages. Our work provides insights into the partnerships in salt marshes and a roadmap for multiscale analyses of diversity in complex biological systems.

Nitrogen removal in constructed marshes at sites protected from and exposed to waves

Nutrient removal is among the most valuable ecosystem services provided by marshes and is often a stated goal of coastal restoration projects. However, the removal capacity of constructed marshes is potentially affected by several site-specific and design factors, such as marsh platform elevation, slope, sediment type, initial planting density and wave climate. Here, the main and interactive effects of these factors on the capacity of constructed marshes to remove nitrate from runoff was explored in field experiments at sites protected from and exposed to waves. At both sites, three experimental blocks were established, each with 24 treatment combinations of factors in experimental flumes: two platform elevations (high and low), two slopes (steep and shallow), two sediment types (coarse and fine grain), and three initial planting densities (0%, 50% and 100% cover). Nutrient rich (KNO3) groundwater solution was fed through the marsh rhizosphere using subsurface diffusers. The relative effects of treatment combinations were then assessed by analyzing porewater NOx concentrations with ANOVA models. None of the treatment combinations had any observable effect on porewater NOx concentrations at the exposed site. However, both sediment type and planting density were significant main effects at the protected site with the lowest NOx concentrations found in flumes with fine sediments and initially planted. These results confirm that design factors can have large implications on nutrient dynamics of constructed marshes in areas protected from waves and that wave energy can substantially reduce the influence of these design factors.

Rhizosphere microbial communities reflect genotypic and trait variation in a salt marsh ecosystem engineer.

There is growing recognition that intraspecific genetic variation in plants can influence associated soil microbial communities, but the functional bridges linking plant genotype with microbial community structure are not well understood. This deficit is due in part to a prevailing focus on characterizing relationships between microbial communities and functional trait variation among plant species or across plant communities, rather than within a single species. We examined whether and how spatiotemporal variation in salt marsh rhizosphere microbial communities reflect plant provenance (genotypic variation) and associated trait variation within an ecosystem engineer, Spartina alterniflora. We planted S. alterniflora from four genetically distinct source populations in replicate sets of experimental plots across a shoreline in southeastern Louisiana, USA. After 2 years, we measured functional plant traits and profiled microbial communities. Bacterial and fungal α-diversity and richness were significantly higher in winter than in summer and corresponded to plant trait variation associated with provenance. Notably, 20% of the variation in fungal community composition was explained by trait differences while bacterial community structure did not reflect plant provenance or trait variation. However, evidence was found suggesting that bacterial communities are indirectly shaped by the influence of plant provenance on soil physicochemical properties. This study illustrates that intraspecific genetic and corresponding trait variation in an ecosystem engineer can shape rhizosphere microbial communities, with fungal communities being more responsive than bacteria to the influence of plant provenance and associated trait variation. Our results highlight the potential relevance of plant intraspecific variation in plant-microbe-soil feedbacks shaping naturally depauperate ecosystems like salt marshes.

Phylogenetic diversity and investigation of plant growth-promoting traits of actinobacteria in coastal salt marsh plant rhizospheres from Jiangsu, China

Actinobacteria from special habitats are of interest due to their producing of bioactive compounds and diverse ecological functions. However, little is known of the diversity and functional traits of actinobacteria inhabiting coastal salt marsh soils. We assessed actinobacterial diversity from eight coastal salt marsh rhizosphere soils from Jiangsu Province, China, using culture-based and 16S rRNA gene high throughput sequencing (HTS) methods, in addition to evaluating their plant growth-promoting (PGP) traits of isolates. Actinobacterial sequences represented 2.8%–43.0% of rhizosphere bacterial communities, as determined by HTS technique. The actinobacteria community comprised 34 families and 79 genera. In addition, 196 actinobacterial isolates were obtained, of which 92 representative isolates were selected for further 16S rRNA gene sequencing and phylogenetic analysis. The 92 strains comprised seven suborders, 12 families, and 20 genera that included several potential novel species. All representative strains were tested for their ability of producing indole acetic acid (IAA), siderophores, 1-aminocyclopropane-1-carboxylate deaminase (ACCD), hydrolytic enzymes, and phosphate solubilization. Based on the presence of multiple PGP traits, two strains, Streptomyces sp. KLBMP S0051 and Micromonospora sp. KLBMP S0019 were selected for inoculation of wheat seeds grown under salt stress. Both strains promoted seed germination, and KLBMP S0019 significantly enhanced seedling growth under NaCl stress. Our study demonstrates that coastal salt marsh rhizosphere soils harbor a diverse reservoir of actinobacteria that are potential resources for the discovery of novel species and functions. Moreover, several of the isolates identified here are good candidates as PGP bacteria that may contribute to plant adaptions to saline soils.

Removal of fast flowing nitrogen from marshes restored in sandy soils.

Groundwater flow rates and nitrate removal capacity from an introduced solution were examined for five marsh restoration designs and unvegetated plots shortly after planting and 1 year post-planting. The restoration site was a sandy beach with a wave-dampening fence 10 m offshore. Simulated groundwater flow into the marsh was introduced at a rate to mimic intense rainfall events. Restoration designs varied in initial planting density and corresponded to 25%, 50%, 75% and 100% of the plot area planted. In general, groundwater flow was slower with increasing planting density and decreased from year 0 to year 1 across all treatments. Nevertheless, removal of nitrate from the introduced solution was similar and low for all restoration designs (3–7%) and similar to the unvegetated plots. We suggest that the low NO3 − removal was due to sandy sediments allowing rapid flow of groundwater through the marsh rhizosphere, thereby decreasing the contact time of the NO3 − with the marsh biota. Our findings demonstrate that knowledge of the groundwater flow regime for restoration projects is essential when nutrient filtration is a target goal of the project.

Microbial Community Analysis of a Coastal Salt Marsh Affected by the Deepwater Horizon Oil Spill

Coastal salt marshes are highly sensitive wetland ecosystems that can sustain long-term impacts from anthropogenic events such as oil spills. In this study, we examined the microbial communities of a Gulf of Mexico coastal salt marsh during and after the influx of petroleum hydrocarbons following the Deepwater Horizon oil spill. Total hydrocarbon concentrations in salt marsh sediments were highest in June and July 2010 and decreased in September 2010. Coupled PhyloChip and GeoChip microarray analyses demonstrated that the microbial community structure and function of the extant salt marsh hydrocarbon-degrading microbial populations changed significantly during the study. The relative richness and abundance of phyla containing previously described hydrocarbon-degrading bacteria (Proteobacteria, Bacteroidetes, and Actinobacteria) increased in hydrocarbon-contaminated sediments and then decreased once hydrocarbons were below detection. Firmicutes, however, continued to increase in relative richness and abundance after hydrocarbon concentrations were below detection. Functional genes involved in hydrocarbon degradation were enriched in hydrocarbon-contaminated sediments then declined significantly (p<0.05) once hydrocarbon concentrations decreased. A greater decrease in hydrocarbon concentrations among marsh grass sediments compared to inlet sediments (lacking marsh grass) suggests that the marsh rhizosphere microbial communities could also be contributing to hydrocarbon degradation. The results of this study provide a comprehensive view of microbial community structural and functional dynamics within perturbed salt marsh ecosystems.

Specificity of Salt Marsh Diazotrophs for Vegetation Zones and Plant Hosts: Results from a North American marsh

Salt marshes located on the east coast of temperate North America are highly productive, typically nitrogen-limited, and support diverse assemblages of free-living nitrogen fixing (diazotrophic) bacteria. This article reviews and analyzes data from North Inlet estuary (SC, USA), addressing diazotroph assemblage structure and the influence of plant host and environmental conditions on the assemblage. The North Inlet estuary is a salt marsh ecosystem in which anthropogenic influences are minimal and the distributions of diazotrophs are governed by the natural biota and dynamics of the system. Denaturing gradient gel electrophoresis fingerprinting and phylogenetic analyses of recovered sequences demonstrated that the distributions of some diazotrophs reflect plant host specificity and that diazotroph assemblages distributed across marsh gradients are also heavily influenced by edaphic conditions. Broadly distributed diazotrophs that are capable of maintaining populations under all environmental conditions spanning such gradients are also present in these assemblages. Statistical analyses indicate that the structures of diazotroph assemblages in different vegetation zones are significantly (p < 0.01) different. New data presented here demonstrate the heterogeneity of salt marsh rhizosphere microenvironments, and corroborate previous findings from different plant hosts growing at several locations within this estuary. The data from these collected works support the hypothesis that the biogeography of microorganisms is non-random and these biogeographic patterns are predictable.

New approach for measuring denitrification in the rhizosphere of vegetated marsh sediments

This study presents a new approach for measuring denitrification at depth in the marsh rhizosphere (below 5 cm). The method combines the push‐pull technique and the 15N‐isotope pairing technique, here referred to as the PPIPT. The PPIPT allows ambient denitrification rates to be measured in situ at depth in the marsh rhizosphere with minimal damage to roots and rhizomes and minimal disturbance of the ambient pore water composition. In this method, pore water is extracted from the sediment using micropiezometers without contact with the atmosphere. The porewater is amended with 15NO3− to trace denitrification, and Ar(g) is added to trace potential gas loss of denitrification end products. The “spiked” pore water is injected back into the sediment for incubation. Subsequently, samples are sequentially extracted from the sediment for analysis of 28N2, 29N2, 30N2, and Ar. Denitrification rates are calculated using the isotope paring technique and corrected for dilution and gaseous losses using the argon tracer. After several trials in the laboratory and in the field to optimize the method, the PPIPT was applied in the field to measure denitrification in the rhizosphere of various vegetation zones in two New England marshes in northeastern USA. The field applications verified the PPIPT method as a useful tool for measuring ambient rates of denitrification at depth in salt marsh rhizospheres. Generally, the rates measured were low (0–12 µmol m−2 h−1) and showed a high spatial variability in the marsh rhizosphere among different vegetation zones.

Salt marsh rhizosphere affects microbial biotransformation of the widespread halogenated contaminant tetrabromobisphenol-A (TBBPA)

Estuarine sediments are the repository for a wide range of contaminants. Anthropogenic impacts and variations in the belowground biomass of salt marsh plants potentially select for different sediment microbial communities with different functional capabilities, including the ability to biotransform anthropogenic contaminants. There are large differences in both root morphology and the amount of fine root biomass of Spartina alterniflora and Phragmites australis; Spartina is the species commonly used to replace Phragmites in northeastern US salt marsh restoration projects. Our study compared the effect of these two macrophyte species on sediment microbial communities responsible for the biotransformation of the halogenated flame retardant tetrabromobisphenol A (TBBPA). Sediments were obtained from contaminated and uncontaminated salt marsh field sites in New Jersey. Anaerobic methanogenic sediment microcosms were established and incubated for up to 130 days. TBBPA was reductively dehalogenated resulting in the transient formation of two intermediates, identified as tribromobisphenol A and dibromobisphenol A, and the formation and accumulation of bisphenol A (BPA) as the end product. Spartina sediments from both sites were found to dehalogenate TBBPA more rapidly than the Phragmites or unvegetated sediments, resulting in greater production of BPA. Microbial community diversity as measured by in situ sediment phospholipid fatty acid (PLFA) composition prior to TBBPA exposure, was found to be higher in the uncontaminated sediments; differences in microbial PLFA diversity were not seen in contaminated sediments associated with either the different plant species or unvegetated sediment. The results of this study demonstrate that these two plant species affected sediment microbial community function with respect to dehalogenation capabilities, even though the disturbed and undisturbed sediments varied in microbial community composition.

A model of biocomplexity and its application to the analysis of some terrestrial and marsh eukaryotic microbial communities with an emphasis on amoeboid protists.

Biocomplexity theory has become increasingly important in understanding ecosystem dynamics as we realize that the interactions among subunits in a multi-component system often produce elaborate states that are not easily explained in terms of the individual parts of the system. A Euclidean geometric model of biocomplexity is presented and illustrated using protistan communities. The model is based on three quantitative biotic dimensions (indices) for small subsamples (0.01 g) taken from each sample core of substratum: (1) richness of morphospecies expressed as mean count per 0.01 g, (2) spatial diversity of protists expressed as then umber of unique morphospecies (i.e. those occurring in only one of the 0.01-g subsamples and not in any of the other subsamples), and (3) patchiness (non-uniform aggregation) of the distribution of protists across the 0.01-g subsamples. These three indices are mapped into a three-dimensional Euclidean space model, and the position of each point and its geometric distance from the origin are used as a general index of biocomplexity. The usefulness of the model is illustrated by applying it to a range of terrestrial and marsh communities. Within the set of 15 samples examined in this study, the marsh rhizosphere samples are among the most complex.

PAH-degradation by Paenibacillus spp. and description of Paenibacillus naphthalenovorans sp. nov., a naphthalene-degrading bacterium from the rhizosphere of salt marsh plants.

Bacteria belonging to the genus Paenibacillus were isolated by enrichment from petroleum-hydrocarbon-contaminated sediment and salt marsh rhizosphere using either naphthalene or phenanthrene as the sole carbon source, and were characterized using phenotypic, morphological and molecular techniques. The isolates were grouped by their colony morphologies and polyaromatic hydrocarbon-degradation patterns. Phenanthrene-degrading isolates produced mottled colonies on solid media and were identified as P. validus by fatty acid methyl ester and 16S rRNA gene sequence analyses. In contrast, the naphthalene-degrading isolates with mucoid colony morphology were distantly related to Paenibacillus validus, according to fatty acid methyl ester and 16S rRNA gene sequence analyses. The predominant fatty acids of the mucoid isolates were 15:0 anteiso, 16:1omega11c, 16:0 and 17:0 anteiso, constituting, on average, 50.5, 12.0, 11.2 and 6.5% of the total, respectively. The G+C contents of their DNA ranged from 47 to 52 mol%. The 16S rDNA sequence analysis revealed the highest (< or = 94%) similarity to P. validus. In addition, phylogenetic analyses based on 16S rDNA sequences showed that the mucoid isolates formed a distinct cluster within Paenibacillus. DNA-DNA hybridization experiments showed only a 6% DNA similarity between the type strain of P. validus and mucoid strain PR-N1. On the basis of the morphological, phenotypic and molecular data, the naphthalene-degrading isolates merit classification as a new Paenibacillus species, for which the name Paenibacillus naphthalenovorans sp. nov. is proposed, with PR-N1T (= ATCC BAA-206T = DSM 14203T) as the type strain.

Isolation and characterization of polycyclic aromatic hydrocarbon-degrading bacteria associated with the rhizosphere of salt marsh plants.

Polycyclic aromatic hydrocarbon (PAH)-degrading bacteria were isolated from contaminated estuarine sediment and salt marsh rhizosphere by enrichment using either naphthalene, phenanthrene, or biphenyl as the sole source of carbon and energy. Pasteurization of samples prior to enrichment resulted in isolation of gram-positive, spore-forming bacteria. The isolates were characterized using a variety of phenotypic, morphologic, and molecular properties. Identification of the isolates based on their fatty acid profiles and partial 16S rRNA gene sequences assigned them to three main bacterial groups: gram-negative pseudomonads; gram-positive, non-spore-forming nocardioforms; and the gram-positive, spore-forming group, Paenibacillus. Genomic digest patterns of all isolates were used to determine unique isolates, and representatives from each bacterial group were chosen for further investigation. Southern hybridization was performed using genes for PAH degradation from Pseudomonas putida NCIB 9816-4, Comamonas testosteroni GZ42, Sphingomonas yanoikuyae B1, and Mycobacterium sp. strain PY01. None of the isolates from the three groups showed homology to the B1 genes, only two nocardioform isolates showed homology to the PY01 genes, and only members of the pseudomonad group showed homology to the NCIB 9816-4 or GZ42 probes. The Paenibacillus isolates showed no homology to any of the tested gene probes, indicating the possibility of novel genes for PAH degradation. Pure culture substrate utilization experiments using several selected isolates from each of the three groups showed that the phenanthrene-enriched isolates are able to utilize a greater number of PAHs than are the naphthalene-enriched isolates. Inoculating two of the gram-positive isolates to a marine sediment slurry spiked with a mixture of PAHs (naphthalene, fluorene, phenanthrene, and pyrene) and biphenyl resulted in rapid transformation of pyrene, in addition to the two- and three-ringed PAHs and biphenyl. This study indicates that the rhizosphere of salt marsh plants contains a diverse population of PAH-degrading bacteria, and the use of plant-associated microorganisms has the potential for bioremediation of contaminated sediments.

Phylogenetic and Physiological Diversity of Sulphate-Reducing Bacteria Isolated from a Salt Marsh Sediment

The phylogenetic and physiological diversity of sulphate-reducing bacteria inhabiting a salt marsh rhizosphere were investigated. Sulphate-reducing bacteria were isolated from a salt marsh rhizosphere using enrichment cultures with electron donors thought to be prevalent in the rhizosphere of Spartina alterniflora. The relationship between phylogeny and nutritional characteristics of 10 strains was investigated. None of the isolates had 16S rRNA sequences identical to other delta subclass sulphate-reducers, sharing 85.3 to 98.1% sequence similarity with 16S rRNA sequences of their respective closest relatives. Phylogenetic analysis placed two isolates, obtained with ethanol as an electron donor, within the Desulfovibrionaceae. Seven isolates, obtained with acetate, butyrate, propionate, or benzoate, were placed within the Desulfobacteriaceae. One isolate, obtained with butyrate, fell within the Desulfobulbus assemblage, which is currently considered part of the Desulfobacteriaceae family. However, due to the phylogenetic breadth and physiological traits of this group, we propose that it be considered a new family, the "Desulfobulbusaceae." The isolates utilised an array of electron donors similar to their closest relatives with a few exceptions. As a whole, the phylogenetic and physiological data indicate isolation of several sulphate-reducing bacteria which might be considered as new species and representative of new genera. Comparison of the Desulfobacteriaceae isolates' 16S rRNA sequences to environmental clones originating from the same study site revealed that none shared more than 86% sequence similarity. The results provide further insight into the diversity of sulphate-reducing bacteria inhabiting the salt marsh ecosystem, as well as supporting general trends in the phylogenetic coherence of physiological traits of delta Proteobacteria sulphate reducers.

Factors affecting short-term variability in sediment pH as a function of marsh elevation in a Virginia mesohaline marsh

An analysis of 5 days of nearly continuously recorded sediment temperature, pH, and radiation measurements along a transect in a mesohaline marsh in Virginia suggests there was a shift in control over short-term pH variability from tidal inundation to radiation with height of the marsh surface at least in the surficial sediments. There was little evidence for tidal control of pH variability at depth in the sediment column. However, biological control of subsurface pH variability was evident both near the tidal creek and in the high marsh. Sediment pH means were generally highest toward the tidal creek and lower in the high marsh. The latter zone had the highest pH variability with diurnal pH excursions up to two units being observed during the experimental period. It is hypothesized that macrophyte transpiration or a series of interlocking mechanisms associated with photosynthesis and microbial activity were responsible for the pH excursions observed at depth in the marsh, since the rapid changes in pH were triggered at sunrise and sunset. The large excursions in sediment pH in the high marsh rhizosphere suggest that geochemical activity may be dynamic over the diurnal cycle.