Large effect of phosphate-solubilizing bacteria on the growth and gene expression of Salix spp. at low phosphorus levels.

Phosphorus is one of the most important nutrients required for plant growth and development. However, owing to its low availability in the soil, phosphorus is also one of the most difficult elements for plants to acquire. Phosphorus released into the soil from bedrock quickly becomes unavailable to plants, forming poorly soluble complexes. Phosphate-solubilizing bacteria (PSB) can solubilize unavailable phosphorus-containing compounds into forms in which phosphorus is readily available, thus promoting plant growth. In this study, two willow species, Salix dasyclados cv. Loden and Salix schwerinii × Salix viminalis cv. Tora, were inoculated with two selected bacterial strains, Pantoea agglomerans and Paenibacillus spp., to evaluate the plant growth parameters and changes in gene expression in the presence of different concentrations of tricalcium phosphate: 0 mM (NP), 1 mM (LP), and 2 mM (HP). Inoculation with PSB increased root, shoot and leaf biomass, and for the HP treatment, significant changes in growth patterns were observed. However, the growth responses to plant treatments tested depended on the willow species. Analysis of the leaf transcriptomes of the phosphate-solubilizing bacterium-inoculated plants showed a large variation in gene expression between the two willow species. For the Tora willow species, upregulation of genes was observed, particularly for those involved in pathways related to photosynthesis, and this effect was strongly influenced by bacterial phosphate solubilization. The Loden willow species was characterized by a general downregulation of genes involved in pathway activity that included ion transport, transcription regulation and chromosomes. The results obtained in this study provide an improved understanding of the dynamics of Salix growth and gene expression under the influence of PSB, contributing to an increase in yield and phosphorus-use efficiency.

Technogenic soil salinisation, vegetation, and management shape microbial abundance, diversity, and activity.

The importance of the microbiome in the functioning of degraded lands in industrialised zones is significant. However, little is known about how environmental parameters affect microbial abundance, structure, diversity, and especially specific guilds involved in the nitrogen cycle in saline soils influenced by the soda industry. To address this knowledge gap, our research focused on assessing the microbiota in relation to soil properties and plant species composition across two transects representing different types of land use: saline wasteland and arable fields. Our findings show that the microbial communities were the most affected not only by soil salinity but also by pH and the composition of plant species. Taxonomic variability was the most shaped by salinity together with management type and CaCO3 content. The impact of salinity on the soil microbiome was manifested in a reduced abundance of bacteria and fungi, a lower number of observed phylotypes, reduced modularity, and a lower abundance of the nitrifying guild. Denitrification and nitrogen fixation were less affected by salinity. The last process was correlated with calcium carbonate. CaCO3 was also associated with microbial taxonomic variability and the overall microbial activity caused by hydrolases, which could aid organic matter turnover in saline but carbonate-rich sites. Bacterial genera such as Bacillus, Peanibacillus, and Rhodomicrobium, in addition to fungal taxa such as Cadophora, Mortierella globalpina, Preussia flanaganii, and Chrysosporium pseudomerdarium, show potential as favourable candidates for possible bioremediation initiatives. These results can be applied to future land reclamation projects. FUNDING INFORMATION: This research received no specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Mixed growth of Salix species can promote phosphate-solubilizing bacteria in the roots and rhizosphere.

Phosphorus (P) is an essential plant nutrient that can limit plant growth due to low availability in the soil. P-solubilizing bacteria in the roots and rhizosphere increase the P use efficiency of plants. This study addressed the impact of plant species, the level of plant association with bacteria (rhizosphere or root endophyte) and environmental factors (e.g., seasons, soil properties) on the abundance and diversity of P-solubilizing bacteria in short-rotation coppices (SRC) of willows (Salix spp.) for biomass production. Two willow species (S. dasyclados cv. Loden and S. schwerinii × S. viminalis cv. Tora) grown in mono-and mixed culture plots were examined for the abundance and diversity of bacteria in the root endosphere and rhizosphere during two seasons (fall and spring) in central Sweden and northern Germany. Soil properties, such as pH and available P and N, had a significant effect on the structure of the bacterial community. Microbiome analysis and culture-based methods revealed a higher diversity of rhizospheric bacteria than endophytic bacteria. The P-solubilizing bacterial isolates belonged mainly to Proteobacteria (85%), Actinobacteria (6%) and Firmicutes (9%). Pseudomonas was the most frequently isolated cultivable bacterial genus from both the root endosphere and the rhizosphere. The remaining cultivable bacterial isolates belonged to the phyla Actinobacteria and Firmicutes. In conclusion, site-specific soil conditions and the level of plant association with bacteria were the main factors shaping the bacterial communities in the willow SRCs. In particular, the concentration of available P along with the total nitrogen in the soil controlled the total bacterial diversity in willow SRCs. A lower number of endophytic and rhizospheric bacteria was observed in Loden willow species compared to that of Tora and the mix of the two, indicating that mixed growth of Salix species promotes P-solubilizing bacterial diversity and abundance. Therefore, a mixed plant design was presented as a management option to increase the P availability for Salix in SRCs. This design should be tested for further species mixtures.

The influence of city development on urban pedodiversity.

The aim of this study was to use a pedodiversity index (PI) to assess changes in the spatial structure of soil cover in Inowroclaw, Poland during the twentieth and twenty-first centuries. An original cartographic approach based on landscape metrics was implemented using GIS techniques and statistical calculations. Based on maps of urban soil complexes and pedodiversity, it was revealed that land and soil cover changes in two studied periods (1934-1978 and 1978-2016) significantly affected pedodiversity in the city. In general, the spatio-temporal increase of the pedodiversity index was observed. The percentage of highest values of the PI ranged from 15.9% in 1934, 17.3% in 1978 to 20.9% in 2016. We revealed that pedodiversity index (PI) values are highly spatially and temporally variable and are associated with urban development and changes in the city's internal structure. The applied approach allowed for the identification of both spatial patterns of changes in soil cover that closely reflect the successive stages of the city's development, and problem areas that require revitalization and the implementation of the principles of sustainable development. Therefore, the proposed method can be recommended for landscape monitoring and in determining ecosystem services in urban and landscape planning, and environmental management.

Can acid sulfate soils from the southern Baltic zone be a source of potentially toxic elements (PTEs)?

The mobility of Cr, Cu, Ni, Pb, and Zn in acid sulfate (AS) soils in the reverse delta of the Swina River, NE Poland was studied using extraction of the target metals using the BCR protocol, which targets acid-extractable (F1), reducible (F2), oxidizable (F3) fractions, and aqua regia digestion for residual (F4) fraction. It was assumed that the content of mobile forms determined in air-dried samples during consequent steps of BCR extraction refers to two scenarios of possible release of selected metals from the studied soils: (1) attributed to seasonal soil moisture variation or (2) caused by artificial drainage. The studied AS soils had thin organic layers (muck, peat and mud) overlaying deltaic sands, and contained hypersulfidic material. The field pH was 6.2 ± 0.5 and significantly decreased to 4.3 ± 1.4 after the 8-weeks incubation period. This can be explained by low buffering properties (e.g. lack of carbonates). Total concentrations of metals (Cr 17.9-61.6, Cu 5.7-27.7, Ni 6.2-47.0, Pb 2.2-17.7, and Zn 13.6-130 mg∙kg-1) in the AS soils were diversified but none of the concentrations exceeded the Polish legal limits. Despite the relative low content of analyzed metals, the studied soils can be a potential source of metal contamination affecting the coastal environment in the southern Baltic Sea region that has been overlooked so far. This is confirmed by the high proportion (44-82%) of Cr, Cu, Ni, and Zn in the F1-F3 fractions which can potentially be released as assumed in two scenarios. The BCR protocol seems to be a useful tool for understanding the chemical behavior and fate of metals in AS soils. Challenges in the assessment of metal mobility in the oxidized and unoxidized zones of individual soil profiles occurred due to their complex morphology resulting from the heterogeneous depositional environment under the human impact.

The Effects of Host Plant Genotype and Environmental Conditions on Fungal Community Composition and Phosphorus Solubilization in Willow Short Rotation Coppice.

Phosphorus (P) is an essential plant nutrient. Low availability of P in soil is mainly caused by high content of Fe2O3 in the clay fraction that binds to P making it unavailable. Beneficial microbes, such as P solubilizing microorganisms can increase the available P in soil and improve plant growth and productivity. In this study, we evaluated the effects of environmental conditions (climate, soil parameters), plant genotype, and level of plant association (rhizosphere or endophytic root organism) on the abundance and diversity of phosphorus solubilizing microorganisms in a Salix production system. We hypothesized that a lower number of endophytic fungi may possess the ability to solubilize P compared to the number of rhizosphere fungi with the same ability. We also expect that the plant genotype and the experimental site with its environmental conditions will influence fungal diversity. Two Salix genotypes grown in pure and mixed cultures were investigated for their fungal microbiome community and diversity in the rhizosphere and endosphere during two growing seasons. We found that the rhizosphere fungal community was more diverse. A general dominance of Ascomycota (Dothideomycetes) and Basidiomycota (Tremellomycetes) was observed. The classes Agaricomycetes and Pezizomycetes were more frequent in the endosphere, while Tremellomycetes and Mortierellomycetes were more abundant in the rhizosphere. Plot-specific soil properties (pH, total organic carbon, and nitrogen) significantly influenced the fungal community structure. Among the culturable fungal diversities, 10 strains of phosphate solubilizing fungi (PSFs) from roots and 12 strains from rhizosphere soil were identified using selective media supplemented with di-calcium and tri-calcium phosphates. The fungal density and the number of PSF were much higher in the rhizosphere than in the endosphere. Penicillium was the dominant genus of PSF isolated from both sites; other less frequent genera of PSFs were Alternaria, Cladosporium, and Clonostachys. Overall the main factors controlling the fungal communities (endophytic vs. rhizosphere fungi) were the soil properties and level of plant association, while no significant influence of growing season was observed. Differences between Salix genotypes were observed for culturable fungal diversity, while in metagenomic data analysis, only the class Dothideomycetes showed a significant effect from the plant genotype.

Transcriptome profiling and environmental linkage to salinity across Salicornia europaea vegetation.

BACKGROUND: Salicornia europaea, a succulent obligatory halophyte is the most salt-tolerant plant species in the world. It survives salt concentrations of more than 1 M. Therefore, it is a suitable model plant to identify genes involved in salt tolerance mechanisms that can be used for the improvement of crops. The changes in a plant's gene expression in response to abiotic stresses may depend on factors like soil conditions at the site, seasonality, etc. To date, experiments were performed to study the gene expression of S. europaea only under controlled conditions. Conversely, the present study investigates the transcriptome and physicochemical parameters of S. europaea shoots and roots from two different types of saline ecosystems growing under natural conditions. RESULTS: The level of soil salinity was higher at the naturally saline site than at the anthropogenic saline site. The parameters such as ECe, Na+, Cl-, Ca+, SO42- and HCO3- of the soils and plant organs significantly varied according to sites and seasons. We found that Na+ mainly accumulated in shoots, whereas K+ and Ca2+ levels were higher in roots throughout the growing period. Moreover, changes in S. europaea gene expression were more prominent in seasons, than sites and plant organs. The 30 differentially expressed genes included enzymes for synthesis of S-adenosyl methionine, CP47 of light-harvesting complex II, photosystem I proteins, Hsp70 gene, ATP-dependent Clp proteases, ribulose bisphosphate carboxylase/oxygenase (Rubisco), phenylalanine ammonia-lyase (PAL), cytochrome c oxidase (COX) and ATP synthase. CONCLUSION: The comparisons made based on two seasons, plant organs and two different sites suggest the importance of seasonal variations in gene expression of S. europaea. We identify the genes that may play an important role in acclimation to season-dependent changes of salinity. The genes were involved in processes such as osmotic adjustment, energy metabolism and photosynthesis.

Bacterial and Fungal Endophytic Microbiomes of Salicornia europaea.

We examined Salicornia europaea, a nonmycorrhizal halophyte associated with specific and unique endophytic bacteria and fungi. The microbial community structure was analyzed at two sites differing in salinization history (anthropogenic and naturally saline site), in contrasting seasons (spring and fall) and in two plant organs (shoots and roots) via 16S rRNA and internal transcribed spacer amplicon sequencing. We observed distinct communities at the two sites, and in shoots and roots, while the season was of no importance. The bacterial community was less diverse in shoot libraries than in roots, regardless of the site and season, whereas no significant differences were observed for the fungal community. Proteobacteria and Bacteroidetes dominated bacterial assemblages, and Ascomycetes were the most frequent fungi. A root core microbiome operational taxonomic unit belonging to the genus Marinimicrobium was identified. We detected a significant influence of the Salicornia bacterial community on the fungal one by means of cocorrespondence analysis. In addition, pathways and potential functions of the bacterial community in Salicornia europaea were inferred and discussed. We can conclude that bacterial and fungal microbiomes of S. europaea are determined by the origin of salinity at the sites. Bacterial communities seemed to influence fungal ones, but not the other way around, which takes us closer to understanding of interactions between the two microbial groups. In addition, the plant organs of the halophyte filter the microbial community composition.IMPORTANCE Endophytes are particularly fascinating because of their multifaceted lifestyle, i.e., they may exist as either free-living soil microbes or saprobic ones or pathogens. Endophytic communities of halophytes may be different than those in other plants because salinity acts as an environmental filter. At the same time, they may contribute to the host's adaptation to adverse environmental conditions, which may be of importance in agriculture.

Bacterial microbiome of root-associated endophytes of Salicornia europaea in correspondence to different levels of salinity.

The halophytes have evolved several strategies to survive in saline environments; however, an additional support from their associated microbiota helps combat adverse conditions. Hence, our driving interests to investigate the endophytic bacterial community richness, diversity, and composition associated to roots of Salicornia europaea from two test sites with different origins of soil salinity. We assumed that salinity will have a negative effect on the diversity of endophytes but simultaneously will permit the high occurrence of halophylic bacteria. Further, to establish the role of the host and its external environment in determining the endophytic diversity, we analyzed the physico-chemical parameters of root zone soil and the concentration of salt ions in the plant roots. The results based on the Miseq Illumina sequencing approach revealed a higher number of endophytic bacterial OTUs at naturally saline test site with a higher level of soil salinity. Proteobacteria and Bacteriodetes were the dominant endophytic phyla at both analyzed sites; additionally, the high occurrence of Planctomycetes and Acidobacteria at more saline site and the occurrence of Firmicutes, Actinobacteria, and Chloroflexi at less saline site were recorded. The salinity in the root zone soil was crucial in structuring the endophytic community of S. europaea, and the significant prevalence of representatives from the phyla Deltaproteobacteria, Acidobacteria, Caldithrix, Fibrobacteres, and Verrucomicrobia at the more saline test site suggest domination of halophylic bacteria with potential role in mitigation of salt stress of halophytes.

How Does Salinity Shape Bacterial and Fungal Microbiomes of Alnus glutinosa Roots?

Black alder (Alnus glutinosa Gaertn.) belongs to dual mycorrhizal trees, forming ectomycorrhizal (EM) and arbuscular (AM) root structures, as well as represents actinorrhizal plants that associate with nitrogen-fixing actinomycete Frankia sp. We hypothesized that the unique ternary structure of symbionts can influence community structure of other plant-associated microorganisms (bacterial and fungal endophytes), particularly under seasonally changing salinity in A. glutinosa roots. In our study we analyzed black alder root bacterial and fungal microbiome present at two forest test sites (saline and non-saline) in two different seasons (spring and fall). The dominant type of root microsymbionts of alder were ectomycorrhizal fungi, whose distribution depended on site (salinity): Tomentella, Lactarius, and Phialocephala were more abundant at the saline site. Mortierella and Naucoria (representatives of saprotrophs or endophytes) displayed the opposite tendency. Arbuscular mycorrhizal fungi belonged to Glomeromycota (orders Paraglomales and Glomales), however, they represented less than 1% of all identified fungi. Bacterial community structure depended on test site but not on season. Sequences affiliated with Rhodanobacter, Granulicella, and Sphingomonas dominated at the saline site, while Bradyrhizobium and Rhizobium were more abundant at the non-saline site. Moreover, genus Frankia was observed only at the saline site. In conclusion, bacterial and fungal community structure of alder root microsymbionts and endophytes depends on five soil chemical parameters: salinity, phosphorus, pH, saturation percentage (SP) as well as total organic carbon (TOC), and seasonality does not appear to be an important factor shaping microbial communities. Ectomycorrhizal fungi are the most abundant symbionts of mature alders growing in saline soils. However, specific distribution of nitrogen-fixing Frankia (forming root nodules) and association of arbuscular fungi at early stages of plant development should be taken into account in further studies.

Ecological risk assessment of heavy metals in salt-affected soils in the Natura 2000 area (Ciechocinek, north-central Poland).

This paper aimed to evaluate the ecological risk posed by the accumulation of heavy metals in the salt-affected soils of the habitat covered by the EU Natura 2000 program in relation to the activity of soil redox enzymes. The research was carried out in the halophyte reserve in Ciechocinek (north-central Poland) which is a very specific habitat as it undergoes a long-term human impact related to both the operation of the medical spa town and the agricultural use of soils in the adjacent areas. The obtained results showed that the content of Zn, Cu, Pb, and Cd in the studied soils exceeded the Polish standards. Based on the obtained data and statistical analysis, it was found that metals may come from two different sources: emission from household boiler rooms (Pb, Cd) and corroded brine sewage pipeline (Zn, Cu).They are characterized by limited mobility due to alkaline environment and strong sorption properties of the clay fraction and organic matter. The correlation analysis indicates that the dehydrogenase activities were negatively correlated with soil electrical conductivity (EC1:5) (r = - 0.665, P < 0.05). Taking into account the protective status of the area, it is difficult to indicate definitely the solution concerning the land management. However, according to the authors, one should pay special attention to a possibility of using halophytes which occur within the reserve for phytoremediation.