Heterogeneity in arbuscular mycorrhizal fungal communities may contribute to inconsistent plant-soil feedback in a Neotropical forest

Background and aims

Plant-soil feedback may vary across host species and environmental gradients. The relative importance of these biotic versus abiotic drivers of feedback will determine the stability of plant and microbial communities across environments. If plant hosts are the main driver of soil microbial communities, plant-soil feedback may be stable across changing environments. However, if microbial communities vary with environmental gradients, feedback may also vary, limiting its capacity to predict plant distributions.

Methods

We characterized arbuscular mycorrhizal (AM) fungi across tree plantations and a primary Neotropical rainforest. We then performed a plant-soil feedback pot experiment of AM fungi from these plantations on three plant species and related feedback and AM fungal communities in the field.

Results

In the field, temporal and spatial variation in AM fungal composition was similar in magnitude to variation across plant host species. Composition of AM fungi in the pot experiment significantly differed from the field plots. Furthermore, differential feedback was explained by shifts in AM fungal composition only for one plant host species (Hyeronima alchorneoides) in the pot experiment.

Conclusions

Natural AM fungal communities were temporally and spatially heterogeneous and AM fungal communities in the greenhouse did not reflect natural soils. These factors led to heterogeneous and unpredictable feedback responses, which suggests that applying greenhouse derived plant-soil feedback trends to predict plant coexistence in natural systems may be misleading.

     

Transient expression of recombinant tissue plasminogen activator (rt-PA) gene in cucurbit plants using viral vector

Objective

To use a transient expression system to express a truncated human tissue plasminogen activator (K2S) gene in cucurbit plants.

Results

The recombinant tissue plasminogen activator protein (K2S form) was expressed in active form in cucurbit plants. Its molecular weight was 43 kDa. The plant-derived rt-PA was determined using goat anti-rabbit antibody by western blotting. Among the infected lines, the highest expression of rt-PA was 62 ng/100 mg per leaf tissue as measured by ELISA. The enzymatic activity of the plant-derived rt-PA was 0.8 IU/ml.

Conclusions

The K25 form of rt-PA was expressed for the first time using the viral expression system. Plant-derived rt-PA showed similar potency to commercially-available PA.

     

Root fungal colonisations of the understory grass <Emphasis Type="Italic">Deschampsia flexuosa</Emphasis> after top-canopy harvesting

Aims

Root fungal relationships in forest understory may be affected by tree harvesting. Deschampsia flexuosa forms a mutualistic symbiosis with arbuscular mycorrhizal (AM) fungi functioning in nutrient uptake, and a more loose association with dark septate endophytic (DSE) fungi. We asked how harvesting affects fungal colonisations and whether DSE is more prone to change than AM.

Methods

Deschampsia flexuosa plants were sampled close to a control or a cut tree after top-canopy harvesting in a primary successional site. Colonisations were studied using light microscopy. Shoot N%, vegetation cover and soil nutrients were determined.

Results

Tree harvesting did not affect vegetation and soil parameters, except potassium (K⁺) increasing near cut trees. AM colonisation did not change, while DSE increased. Shoot N% increased with increasing DSE near cut trees. Hyaline septate (HSE) hyphae and soil K⁺ and magnesium (Mg²⁺) were positively correlated near control trees. Lichen cover and HSE correlated negatively.

Conclusions

DSE colonisation increased but AM did not change after harvesting. Positive correlation of DSE with shoot N% near cut trees may suggest a role for DSE in favouring plant nitrogen uptake after disturbance in an open microsite. HSE may play a role in K⁺ and Mg²⁺ uptake.

     

Variation of soil respiration and its components in hemiboreal Norway spruce stands of different ages

Aims

Understanding the linkage of soil respiration (Rs) with forest development is essential for long-term C cycle models. We estimated the variation and temperature sensitivity (Q₁₀ value) of Rs and its hetero-, (Rh) and autotrophic (Ra) components in relation to abiotic and biotic factors in Norway spruce stands of different ages, and the effect of trenching on microbial and soil characteristics.

Methods

Trenching method was used to partition Rs into Rh and Ra. Ingrowth core method was used to estimate fine root production. Soil microbial biomass was measured using manometric respirometers.

Results

Rs varied in differently aged stands demonstrating non-linear response to development stage. The variation of Rs was explained by changes in biotic factors rather than by changes in soil microclimate. Rh was more sensitive to Ts than Rs or Ra. After 4 years of trenching soil pH, N, SOM and dehydrogenase activity were significantly changed in trenched plots compared to control plots.

Conclusions

Different Q₁₀ values of Rh and Ra in stands of different ages indicate the importance of Rs partitioning. Trenching should be used during a limited number of years because of the possible changes in chemical characteristics of soil and in the activity of soil microbial community.

     

Roles of shoots and roots in cadmium uptake and distribution in tubers of potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L)

Aims

We investigated whether density fractionation can be used to determine the distribution of organic phosphorus (OP) between free and mineral-associated soil organic matter (SOM).

Methods

We performed density fractionations using sodium polytungstate solution (specific gravity 1.6 g cm^?3) on 20 soils from UK semi-natural and pasture ecosystems, to obtain a light fraction (LF) and a heavy fraction (HF) for each soil. The fractions were quantified by weight, and analysed for organic carbon (OC), total N (TN), total P (TP), inorganic P (IP), and OP (by difference).

Results

Good recoveries of soil mass (96%), OC and TN (both ~ 90%) were obtained, but recovery of OP only averaged 56%. The average P:C ratio of HF SOM exceeded that of LF SOM by a factor of six, greater than the factor of two obtained for TN:OC. For the soils studied, the elements of SOM were predominantly in the HF, with averages of 75% for C, 82% for N, and 90% for P.

Conclusions

The incomplete recovery of OP demands further work. Nonetheless, the results show that HF SOM is much richer in P than LF SOM.

     

The bacterial community associated with rose-scented geranium (<Emphasis Type="Italic">Pelargonium graveolens</Emphasis>) leaves responds to anthracnose symptoms

Background

The fungus Colletotrichum is a plant pathogen that causes the anthracnose disease, resulting in huge losses in various crops including the rose-scented geranium (Pelargonium graveolens). Although the bacterial community associated with plants has an important role in the establishment of plant diseases, little is known about what happens in P. graveolens.

Aims

To increase the knowledge about the bacterial community associated with P. graveolens and its relationship with anthracnose disease symptoms.

Methods

Quantitative PCR and high-throughput sequencing were combined to determine the presence of the fungus Colletotrichum and to reveal the bacterial communities associated with different plant parts – root, stem and leaf – and in the rhizosphere and bulk soil, and also to determine the respective bacterial communities associated with P. graveolens leaves symptomatic and asymptomatic for anthracnose disease.

Results

The fungus Colletotrichum was detected in all plant parts and in the surrounding soil. Bacterial communities varied spatially in plants, and the disease symptoms also influenced the composition of the bacterial community. Abundances of operational taxonomic units (OTUs) assigned to the phylum Actinobacteria and to the genus Streptococcus were greatly increased in asymptomatic leaves.

Conclusions

The bacterial community associated to geranium leaves responds to anthracnose symptoms.

     

Chemical changes in organic matter after fungal colonization in a nitrogen fertilized and unfertilized Norway spruce forest

Background and aims

Mineralization of soil organic matter (SOM) constitutes a major carbon flux to the atmosphere. The carbon use efficiency (CUE) of the saprotrophic microorganisms mineralizing SOM is integral for soil carbon dynamics. Here we investigate how the CUE is affected by temperature, metabolic conditions, and the molecular complexity of the substrate.

Methods

We incubated O-horizon soil samples (with either ¹³C–glucose or ¹³C–cellulose) from a boreal coniferous forest at 4, 9, 14, and 19 °C, and calculated CUEs based on the amount of ¹³C–CO₂ and ¹³C–labelled microbial biomass produced. The effects of substrate, temperature, and metabolic conditions (representing unlimited substrate supply and substrate limitation) on CUE were evaluated.

Results

CUE from metabolizing glucose was higher as compared to cellulose. A slight decrease in CUE with increasing temperature was observed in glucose amended samples (but only in the range 9–19 °C), but not in cellulose amended samples. CUE differed significantly with metabolic conditions, i.e. CUE was higher during unlimited growth conditions as compared to conditions with substrate limitation.

Conclusions

We conclude that it is integral to account for both differences in CUE during different metabolic phases, as well as complexity of substrate, when interpreting temperature dependence on CUE in incubation studies.

     

Trade-off between C and N recycling and N<Subscript>2</Subscript>O emissions of soils with summer cover crops in subtropical agrosystems

Aims

Biological soil crusts (biocrusts) are soil-surface communities in drylands, dominated by cyanobacteria, mosses, and lichens. They provide key ecosystem functions by increasing soil stability and influencing soil hydrologic, nutrient, and carbon cycles. Because of this, methods to reestablish biocrusts in damaged drylands are needed. Here we test the reintroduction of field-collected vs. greenhouse-cultured biocrusts for rehabilitation.

Methods

We collected biocrusts for 1) direct reapplication, and 2) artificial cultivation under varying hydration regimes. We added field-collected and cultivated biocrusts (with and without hardening treatments) to bare field plots and monitored establishment.

Results

Both field-collected and cultivated cyanobacteria increased cover dramatically during the experimental period. Cultivated biocrusts established more rapidly than field-collected biocrusts, attaining ~82% cover in only one year, but addition of field-collected biocrusts led to higher species richness, biomass (as assessed by chlorophyll a) and level of development. Mosses and lichens did not establish well in either case, but late successional cover was affected by hardening and culture conditions.

Conclusions

This study provides further evidence that it is possible to culture biocrust components from later successional materials and reestablish cultured organisms in the field. However, more research is needed into effective reclamation techniques.

     

Interaction between contrasting rice genotypes and soil physical conditions induced by hydraulic stresses typical of alternate wetting and drying irrigation of soil

Background and aims

Drought events, agricultural practices and plant communities influence microbial and soil abiotic parameters which can feedback to fodder production. This study aimed to determine which soil legacies influence plant biomass production and nutritional quality, and its resistance and recovery to extreme weather events.

Methods

In a greenhouse experiment, soil legacy effects on Lolium perenne were examined, first under optimal conditions, and subsequently during and after drought. We used subalpine grassland soils previously cultivated for two years with grass communities of distinct functional composition, and subjected to combinations of climatic stress and simulated management.

Results

The soil legacy of climatic stress increased biomass production of Lolium perenne and its resistance and recovery to a new drought. This beneficial effect resulted from higher nutrient availability in soils previously exposed to climatic stresses due to lower competitive abilities and resistance of microbial communities to a new drought. This negative effect on microbial communities was strongest in soils from previously cut and fertilized grasslands or dominated by conservative grasses.

Conclusion

In subalpine grasslands more frequent climatic stresses could benefit fodder production in the short term, but threaten ecosystem functioning and the maintenance of traditional agricultural practices in the long term.

     

Differential elemental uptake in three pseudo-metallophyte C<Subscript>4</Subscript> grasses in situ in the eastern USA

Background and aims

Elemental uptake in serpentine floras in eastern North America is largely unknown. The objective of this study was to determine major and trace element concentrations in soil and leaves of three native pseudo-metallophyte C₄ grasses in situ at five sites with three very different soil types, including three serpentine sites, in eastern USA.

Methods

Pseudo-total and extractible concentrations of 15 elements were measured and correlated from the soils and leaves of three species at the five sites.

Results

Element concentrations in soils of pseudo-metallophytes varied up to five orders of magnitude. Soils from metalliferous sites exhibited higher concentrations of their characteristic elements than non-metalliferous. In metallicolous populations, elemental concentrations depended on the element. Concentrations of major elements (Ca, Mg, K) in leaves were lower than typical toxicity thresholds, whereas concentrations of Zn were higher.

Conclusions

In grasses, species can maintain relatively low metal concentrations in their leaves even when soil concentrations are richer. However, in highly Zn-contaminated soil, we found evidence of a threshold concentration above which Zn uptake increases drastically. Finally, absence of main characteristics of serpentine soil at one site indicated the importance of soil survey and restoration to maintain serpentinophytes communities and avoid soil encroachment.

     

Soil under nurse plants is always better than outside: a survey on soil amelioration by a complete guild of nurse plants across a long environmental gradient

Aims

Soil under nurse plants is more fertile than in the harsh surroundings. This is a primary mechanism involved in plant to plant facilitation and it is critical in structuring plant communities under stressful conditions. However we do not know how this soil enrichment process varies along complex environmental gradients and among coexisting nurse plants.

Methods

Soil properties related to structure, resource stocks and microbial activity, were compared among up to ten nurse plant species and adjacent barren soil areas, along a 1600 m elevation gradient above the treeline in central Chilean Andes. Shifts in Relative Interaction Index (RII) sensu Armas (Ecology 85: 2682–2686, 2004) and in coefficient of variation on soil properties were also modelled.

Results

Soil under nurse plants was always richer than on barren areas irrespective of altitude, except in the case of texture with more small particles in the intermediate altitude. β-glucosidase activity was higher under cushion plants than under nurse plants with other growth habit. Besides β-glucosidase and phosphatase activities were more variable at higher altitudes. Nitrogen was more variable under nurse plants than in barren areas and its RII values were lower at intermediate altitudes.

Conclusions

Soil amelioration by nurse plants occurred all along the studied environmental gradient promoting islands of fertility and a general increase on soil niches heterogeneity.

     

Validating novel oligonucleotide primers targeting three classes of bacterial non-specific acid phosphatase genes in grassland soils

Background and aims

Microbially driven mineralization of organic phosphorus forms is of particular importance in the soil environment, where it becomes available to plants as inorganic orthophosphates. In acidic soils, microbes produce non-specific acid phosphatases (NSAPs; E.C. 3.1.3.2) which act on the most common forms of organic P in the soil. Our understanding of phosphorus turnover in soils would greatly benefit from an improvement in research tools targeting these genes.

Methods

Thus, in this study we developed two novel oligonucleotide PCR primer sets, that will enable researchers to target the present and active communities of bacteria with the genetic potential of acid phosphatase production. A total of three primer sets were validated to target the three classes of NSAPs. Utilizing Illumina MiSeq, amplicons from grassland pasture soils were sequenced.

Results

The resulting target specificity was high for all three groups; CAAP (97.2%), CBAP (99.5%), and CCAP (94.8%). Quantification of target genes by qPCR indicated measurable differences between classes, ranging from 5 log to 7.5 log for CAAP, 6 log to 8 log for CBAP, and 4 log to 5 log for CCAP.

Conclusions

The validated primer sets were specific to the target genes and identified potential quantitative differences between the NSAP classes.

     

RiboTagger: fast and unbiased 16S/18S profiling using whole community shotgun metagenomic or metatranscriptome surveys

Background

Taxonomic profiling of microbial communities is often performed using small subunit ribosomal RNA (SSU) amplicon sequencing (16S or 18S), while environmental shotgun sequencing is often focused on functional analysis. Large shotgun datasets contain a significant number of SSU sequences and these can be exploited to perform an unbiased SSU--based taxonomic analysis.

Results

Here we present a new program called RiboTagger that identifies and extracts taxonomically informative ribotags located in a specified variable region of the SSU gene in a high-throughput fashion.

Conclusions

RiboTagger permits fast recovery of SSU-RNA sequences from shotgun nucleic acid surveys of complex microbial communities. The program targets all three domains of life, exhibits high sensitivity and specificity and is substantially faster than comparable programs.

     

A lost opportunity for science: journals promote data sharing in metabolomics but do not enforce it

Introduction

Data sharing is being increasingly required by journals and has been heralded as a solution to the ‘replication crisis’.

Objectives

(i) Review data sharing policies of journals publishing the most metabolomics papers associated with open data and (ii) compare these journals’ policies to those that publish the most metabolomics papers.

Methods

A PubMed search was used to identify metabolomics papers. Metabolomics data repositories were manually searched for linked publications.

Results

Journals that support data sharing are not necessarily those with the most papers associated to open metabolomics data.

Conclusion

Further efforts are required to improve data sharing in metabolomics.

     

Eighty years of maize breeding alters plant nitrogen acquisition but not rhizosphere bacterial community composition

Background and aims

Phosphorus (P) availability is crucial for forest ecosystem productivity and soil organic matter (SOM) is an important source for P. This study was conducted to reveal carbon (C), nitrogen (N) and P distributions in functional SOM fractions. We hypothesised that (1) most of the organic P (P_org) is part of the particulate SOM, (2) particulate SOM stores increasing share of P with decreasing soil P content and (3) the C:P_org ratio of mineral-associated SOM is smaller than that of particulate SOM.

Methods

We analysed soil samples from five temperate forest sites (Fagus sylvatica) under different geological parent material with a wide range of total P concentrations. Density fractionation was used to separate free light fraction (fLF), particulate SOM occluded within soil aggregates (occluded light fraction; oLF), and mineral associated SOM (heavy fraction; HF). We determined the mass balance of P in these fractions, in addition to the C and N concentrations. Additionally, the P speciation of the topsoil was analysed by X-ray absorption near edge structure (XANES) spectroscopy at the P K-edge.

Results

The fLF contained 18–54% and the oLF 1–15% of total P (P_tot). High percentage of P in these light fractions was associated to soil minerals. Phosphorous in particulate SOM within aggregates tend to increase with decreasing soil P. The HF containing mineral-associated OM, comprised 38–71% of P_tot and their C:P_org ratios were consistently lower than those of the fLF irrespective of the P status of the soil.

Conclusions

We show that all three functional SOM fractions contain variable amount of both organic and inorganic P species. The free light fraction shows no response to changing P stocks of soils.. Despite physically protected particulate SOM, oLF, becomes increasingly relevant as P cache in soils with declining P status.

     

Soil micro-food web interactions and rhizosphere priming effect

Background

Seeds host bacterial inhabitants but only a limited knowledge is available on which taxa inhabit seed, which niches could be colonized, and what the routes of colonization are.

Scope

Within this commentary, a discussion is provided on seed bacterial inhabitants, their taxa, and from where derive the seed colonizers.

Conclusions

Seeds/and grains host specific bacteria deriving from the anthosphere, carposphere, or from cones of gymnosperms and inner tissues of plants after a long colonization from the soil to reproductive organs.

     

Sample storage conditions alter colonisation structures of arbuscular mycorrhizal fungi and,particularly, fine root endophyte

Background and Aims

The structures of arbuscular mycorrhizal (AM) fungi (hyphae, arbuscules, vesicles, spores) are used to make inferences about fungal activity based on stored samples, yet the impact of storage method has not been quantified, despite known effects of temperature and host condition on AM fungal colonisation.

Methods

We measured how four storage treatments (cool or ambient conditions, with and without plant shoots attached, i.e. n?=?four treatment combinations) affected AM fungal colonisation of subterranean clover (Trifolium subterraneum L.) after 0, 2, 6 and 10 days of storage. Roots were assessed for colonisation of fine root endophyte and coarse AM fungi.

Results

For coarse AM fungi, total colonisation was unaffected, but arbuscules were reduced at Day 6 and increased again by Day 10, except Ambient-Minus-Shoots. There was a loss of vesicles in all treatments at Day 2, and an increase in spore number at Day 6 within Cool-Plus-Shoots. In contrast, for fine root endophyte, total colonisation was greatly reduced at Day 6 but increased again at Day 10, in all except the Cool-Plus-Shoots treatment.

Conclusions

Our data demonstrate that AM fungal activity is not suspended in commonly used plant storage conditions. Storage method and time impacted AM fungal colonisation, particularly for fine root endophyte. We recommend samples are processed within 2 days of harvest.

     

Arbuscular mycorrhizal fungi can shift plant-soil feedback of grass-endophyte symbiosis from negative to positive

Background and aims

Variations in root-associated fungal communities contribute to the so-called ‘crop rotation benefit’ on soil productivity. We assessed the effects of chickpea, lentil, and pea in wheat-based rotations, as compared to wheat monoculture, on the structure of root-associated fungal communities, and described the legacy of pulses on a following wheat crop.

Methods

The internal transcribed spacer (ITS) and 18S rRNA gene markers, and 454 amplicon pyrosequencing were used to describe the fungal communities of crop roots and rhizosphere soil in a field experiment and agronomic data were collected.

Results

Pulses influenced only the structure of the non-mycorrhizal fungal community of roots. Fusarium tricinctum, Clonostachys rosea, Fusarium redolens, and Cryptococcus sp. were specific to certain crops. Despite the absence of selective effects of pulses on their associated arbuscular mycorrhizal (AM) fungal community, pea had a legacy effect on the structure of the AM fungal community associated with the roots of the following wheat crop, in one of the two year/sites examined. Species of Mortierella, Cryptococcus, and Paraglomus in wheat rhizosphere soil may benefit yield, whereas species of Fusarium, Davidiella, Lachnum, Sistotrema and Podospora may reduce yield.

Conclusion

The effect of pulse crops on root fungal communities varied with rotation crop species. Pulses had various effects on the physiology of the following wheat crop, including increased productivity.

     

Molecular pharming’s foot in the FDA’s door: Protalix’s trailblazing story

Objectives

This short commentary examines the factors that led to Food and Drug Administration’s approval of the first plant-derived biologic.

Results

In 2012, the first plant-derived protein pharmaceutical (biologic) was approved for commercial use in humans. The product, a recombinant form of human β-glucocerebrosidase marketed as ELELYSO, was developed by Protalix Biotherapeutics (Carmiel, Israel). The foresight to select this particular therapeutic product for development, flawless production pipeline, and serendipity seem to provide the key in explaining how ELELYSO became the first plant-derived biologic to achieve approval by Food and Drug Administration.

Conclusions

While the circumstances that enabled Protalix and its scientists to become the first to arrive at this historic milestone are perhaps unique, it is anticipated that more biologics will follow suit in winning regulatory endorsement.

     

Dynamics of plant nutrient uptake as affected by biopore-associated root growth in arable subsoil

Background and aims

Soil microbial communities influence nutrient cycling, chemistry and structure of soil, and plant productivity. In turn, agronomic practices such as fertilization and crop rotation alter soil physical and chemical properties and consequently soil microbiomes. Understanding the long-term effects of agronomic practices on soil microbiomes is essential for improving agronomic practices to optimize these microbial communities for agricultural sustainability. We examine the composition and substrate-utilization profiles of microbial communities at the Morrow Plots in Illinois.

Methods

Microbial community composition is assessed with 16S rRNA gene sequencing and subsequent bioinformatic analyses. Community- level substrate utilization is characterized with the BIOLOG EcoPlate.

Results

Fertilizer and rotation treatments significantly affected microbial community structure, while substrate utilization was affected by fertilizer, but not crop-rotation treatments. Differences in relative abundance and occurrence of bacterial taxa found in fertilizer treatments can explain the observed differences in community level substrate utilization.

Conclusion

Long-term fertilization and crop-rotation treatments affect soil microbial community composition and physiology, specifically through chronic nutrient limitation, long-term influx of microbes and organic matter via manure application, as well as through changes in soil chemistry. Relatively greater abundance of Koribacteraceae and Solibacterales taxa in soils might prove useful as indicators of soil degradation.