Natural ageing of biochar improves its benefits to soil Pb immobilization and reduction in soil phytotoxicity.

Amendments are good tools for immobilizing metal(loid) and improving phytoremediation success. However, the amendment effect is variable and depends on multiple parameters, including amendment type and ageing. Such an ageing effect is rarely studied. Our study is one of the first focusing on how biochar storage affects its effect on soil properties and metal(loid) immobilization, when biochar was applied alone or in combination with green manure. To answer this, a 33-day pot incubation experiment was set up using contaminated soil, amended with two biochars (differing in ages: old (Bo) and new (Bn)) and/or two green manures (leaves of clover or poplar) and sown with Phaseolus vulgaris (bioindicator plant). Soil pore waters, plant growth and metal(loid) accumulation were evaluated. Biochar reduced soil acidity (Bn: + 0.75 pH unit, Bo: + 0.72 unit) and Pb mobility (Bn: - 42%, Bo: - 50%), while green manures acidified the soil (- 0.30 pH unit) and immobilized Pb only after 10 days (- 44%). All amendments reduced soil phytotoxicity. Moreover, the biochar stored at room temperature for a few years demonstrated better abilities to improve soil properties, particularly for Pb immobilization, than the biochar freshly prepared. Finally, as mixtures maturated, soil parameters changed until about ten days, then tended to stabilize. Therefore, it can be concluded that (1) biochar storage will affect its chemical properties and ameliorate its effects, (2) biochar can ameliorate soil properties and immobilize metal(loid)s, while green manures tended to have adverse effects at first, and (3) soil/amendment mixtures should be left to mature about two weeks before potential plant implementation.

Root and shoot biology of Arabidopsis halleri dissected by WGCNA: an insight into the organ pivotal pathways and genes of an hyperaccumulator.

Arabidopsis halleri is a hyperaccumulating pseudo-metallophyte and an emerging model to explore molecular basis of metal tolerance and hyperaccumulation. In this regard, understanding of interacting genes can be a crucial aspect as these interactions regulate several biological functions at molecular level in response to multiple signals. In this current study, we applied a weighted gene co-expression network analysis (WGCNA) on root and shoot RNA-seq data of A. halleri to predict the related scale-free organ specific co-expression networks, for the first time. A total of 19,653 genes of root and 18,081 genes of shoot were grouped into 14 modules and subjected to GO and KEGG enrichment analysis. "Photosynthesis" and "photosynthesis-antenna proteins" were identified as the most enriched and common pathway to both root and shoot. Whereas "glucosinolate biosynthesis," "autophagy," and "SNARE interactions in vesicular transport" were specific to root, and "circadian rhythm" was found to be enriched only in shoot. Later, hub and bottleneck genes were identified in each module by using cytoHubba plugin based on Cytoscape and scoring the relevance of each gene to the topology of the network. The modules with the most significant differential expression pattern across control and treatment (Cd-Zn treatment) were selected and their hub and bottleneck genes were screened to validate their possible involvement in heavy metal stress. Moreover, we combined the analysis of co-expression modules together with protein-protein interactions (PPIs), confirming some genes as potential candidates in plant heavy metal stress and as biomarkers. The results from this analysis shed the light on the pivotal functions to the hyperaccumulative trait of A. halleri, giving perspective to new paths for future research on this species.

The Challenges of Diagnosis and Treatment of Arrhythmogenic Cardiomyopathy: Are We there yet?

Background: we sought to review the evolution in the diagnosis and treatment of Arrhythmogenic Cardiomyopathy (ACM), a clinically multifaceted entity beyond the observation of ventricular arrhythmias, and the outcome of therapies aiming at sudden death prevention in a single center experience. Methods: retrospective analysis of the data of consecutive patients with an implanted cardioverter-defibrillator (ICD) and a confirmed diagnosis of ACM according to the proposed Padua Criteria, who were referred to our center from January 1992 to October 2021. Results: we enrolled 72 patients (66% males, mean age at implant 46 ± 16 years), 63.9% implanted for primary prevention. At the time of ICD implant, 29 (40.3%) patients had a right ventricular involvement, 24 (33.3%) had a dominant LV involvement and 19 (26.4%) had a biventricular involvement. After a median follow-up of 6,1 years [IQR: 2.5-9.9], 34 patients (47.2%) had 919 sustained episodes of ventricular arrhythmias (VA). 27 patients (37.5%) had 314 episodes of life-threatening arrhythmias (LT-VA), defined as sustained ventricular tachycardia ≥ 200 beats/min. Considering only the patients with an ICD capable of delivering ATP, 80.4% of VA and 65% of LT-VA were successfully terminated with ATP. 16 (22.2%) patients had an inappropriate ICD activation, mostly caused by atrial fibrillation, while in 9 patients (12.5%) there was a complication needing reintervention (in 3 cases there was a loss of ventricular sensing dictating lead revision). During the follow-up 11 (15.3%) patients died, most of them due to heart failure, and 8 (11.1%) underwent heart transplantation. Conclusions: ACM is increasingly diagnosed owing to heightened suspicion at ECG examination and to improved imaging technology and availability, though the diagnostic workflow is particularly challenging in the earliest disease stages. ICD therapy is the cornerstone of sudden death prevention, albeit its efficacy is not based on controlled studies, and VT ablation/medical therapy are complementary to this strategy. The high burden of ATP-terminated VA makes shock-only devices debatable. The progressive nature of ACM leads to severe biventricular enlargement and refractory heart failure, which pose significant treatment issues when a predominant RV dysfunction occurs owing to the reduced possibility for mechanical circulatory assistance.

Co-culture of Salix viminalis and Trifolium repens for the phytostabilisation of Pb and As in mine tailings amended with hardwood biochar.

Metal(loid) soil pollution causes environmental and health issues, and thus those sites need to be remediated. This can be done through phytostabilization, in combination with biochar amendment. The objectives were to investigate the potential of Salix viminalis L. associated with Trifolium repens L. for the phytostabilization of biochar-amended contaminated soils by assessing (1) the tolerance of both plants to metal(loid)s, through the biomass production, (2) the concentrations of metal(loid)s in plant parts and (3) the concentrations of metal(loid)s in soil pore water and percolation waters. Results showed that plant growth affected soil pore water Physico-chemical properties and metal(loid) mobility. When comparing the mono- and poly-cultures, although pH was higher with the polyculture than the monoculture, the decrease in Pb mobility did not differ. Moreover, the leachate analysis showed that As concentration in the soil particles leached from the soil was higher in the polyculture condition, while Pb concentration was the highest in the willow vegetated condition. Finally, willow dry weight was not affected by the presence of clover, while clover dry weight was lower when it was grown with willow. In conclusion, the results showed that the willow and clover polyculture was not better than the monoculture of these two species for the phytomanagement of a former mine site amended with biochar.

Effects of Different Biochars, Activated Carbons and Redmuds on the Growth of Trifolium repens and As and Pb Stabilization in a Former Mine Technosol.

Soil pollution by metal(loid)s is an important issue in Europe, as it causes environmental and health problems. Therefore, remediation of these areas is needed. The success of phytoremediation process will depend on the ability of plants to implement, which can require the addition of amendments to the soil in order to improve soil conditions, immobilize pollutant and thus ameliorate plant growth. Amendments that can be used are biochar, activated carbon and redmuds, all of which have previously shown positive outcomes. The objectives of this study were to evaluate the effects of several amendments (biochars, activated carbons and redmuds) on (i) the soil physico-chemical properties of a former mine technosol contaminated by As and Pb, (ii) As and Pb immobilization and (iii) the growth of Trifolium repens. Results showed that amendment addition could ameliorate soil conditions, by reducing soil acidity (pH increased by 1.2 to 1.7 units) and immobilizing pollutants (85 to 99% of Pb immobilized); and improve plant growth (dry weight increased 1.5 to 2.5 times). However, not all amendments were beneficial to the soil and plant. For instance, the L27 activated carbon acidified soil pH, mobilized As and lowered plant growth. This study has allowed us to conclude that amendment effect is dependent on soil type, metal(loid)s and amendment properties, and it is thus necessary to choose the right amendment. Finally, amendments could be combined for better outcomes.

Effect of biochar, iron sulfate and poultry manure application on the phytotoxicity of a former tin mine.

In phytomanagement approach the application of a combination of amendments is an option for remediating arsenic polluted areas and valorized biomass obtained. Various amendments can be used. Biochar has been shown to reduce metal(loid) availability, and increase soil fertility, while iron sulfate has a considerable As binding capacity, and poultry manure is a source of nutrients. A phytotoxicity test was performed by applying the three amendments (2% biochar, 0.15%, 0.30% and 0.45% iron sulfate and 0.4% poultry manure) to a former tin mine technosol, to investigate their effects on (i) soil pore water properties, (ii) metal(loid) immobilization and (iii) Phaseolus vulgaris L. growth, used as a bioindicator. Biochar addition alone did not affect soil properties or plant parameters. However, the addition of iron sulfate acidified the soil, decreased soil pore water As concentrations, and increased the ones of Fe and Pb. It also improved plant growth, and reduced As and Pb aerial and root concentrations. Finally, the addition of poultry manure had no effect on soil and plants. Based on our results, the combination of iron sulfate with biochar may be a solution for reducing soil toxicity of the Abbaretz mining technosol, improving its fertility, and thus ameliorating plant growth.Novelty statement:The work presented in this manuscript describes the effect of amendment application, i.e., biochar, chicken dung and/or iron sulfate, on soil properties, metals availability and dwarf bean growth, plant used as bioindicator.Our results showed that the combination of a low amount of iron sulfate with biochar is the strategy to reduce soil toxicity, improved its fertility and consequently authorizes plant growth.This study is one of the first describing the effects of combined amendments on a mining soil properties with focusing on metal(loid) mobility.

Contrasted tolerance of Agrostis capillaris metallicolous and non-metallicolous ecotypes in the context of a mining technosol amended by biochar, compost and iron sulfate.

Metal(loid) contamination of soil, resulting from the mining activities, is a major issue worldwide, due to its negative effects on the environment and health. Therefore, these contaminated soils need to be remediated. One realistic method is the assisted phytostabilization, which aims at establishing a vegetation cover on the soil that will reduce metal(loid) bioavailability and spreading through the prevention of wind erosion and water leaching. In addition, amendments are applied to improve soil conditions and ameliorate plant growth. In this goal, biochar and compost showed good results in terms of amelioration of soil fertility and reduction in lead bioavailability. However, they usually have a negative effect on arsenic. On the contrary, iron sulfate showed capacity to reduce arsenic mobility through interaction with its iron hydroxides. Finally, the choice of the appropriate plant species is crucial for the success of assisted phytostabilization. One good option is to use endemic species, adapted to the metal(loid) stress, with a fast growth and large shoot and root systems. The aims of this study were to (1) evaluate the effects of applying biochar, compost and iron sulfate, alone or combined, to a former mine soil on the soil properties and Agrostis capillaris growth, and (2) assess the difference between two Agrostis capillaris ecotypes, an endemic metallicolous ecotype and a non-metallicolous ecotype. Results of the mesocosm experiment showed that amendment application improved soil properties, i.e., reduced soil acidity, increased nutrient availability and lower metal(loid) stress, the best being the combination biochar-compost-iron sulfate. These ameliorations allowed a better plant growth. Finally, the metallicolous ecotype performed better in terms of growth than the non-metallicolous one and could thus be used in an assisted phytostabilization process on the former mine site.

Application of amendments for the phytoremediation of a former mine technosol by endemic pioneer species: alder and birch seedlings.

Metal(loid) pollution of soils has important negative effects on the environment and human health. For the rehabilitation of these soils, some eco-innovative strategies, such as phytoremediation, could be chosen. This practice could establish a plant cover to reduce the toxicity of the pollutants and stabilize the soil, preventing soil erosion and water leaching; this technique is called phytoremediation. For this, plants need to be tolerant to the pollutants present; thus, phytoremediation can have better outcomes if endemic species of the polluted area are used. Finally, to further improve phytoremediation success, amendments can be applied to ameliorate soil conditions. Different amendments can be used, such as biochar, a good metal(loid) immobilizer, compost, a nutrient-rich product and iron sulfate, an efficient arsenic immobilizer. These amendments can either be applied alone or combined for further positive effects. In this context, a mesocosm experiment was performed to study the effects of three amendments, biochar, compost and iron sulfate, applied alone or combined to a former mine technosol, on the soil properties and the phytoremediation potential of two endemic species, Alnus sp. and Betula sp. Results showed that the different amendments reduced soil acidity and decreased metal(loid) mobility, thus improving plant growth. Both species were able to grow on the amended technosols, but alder seedlings had a much higher growth compared to birch seedlings. Finally, the combination of compost with biochar and/or iron sulfate and the establishment of endemic alder plants could be a solution to rehabilitate a former mine technosol.

Effects of biochar, ochre and manure amendments associated with a metallicolous ecotype of Agrostis capillaris on As and Pb stabilization of a former mine technosol.

Metal(loid) soil pollution is a major environmental and health issue, requiring these areas to be remediated, for example through phytoremediation processes. In order to allow proper plant establishment and growth, amendments must be applied to highly contaminated and poorly fertile soils. Amendments are diverse, but many studies have shown the beneficial effects of biochar, manure and ochre, although studies on their combined use are scarce. Moreover, no studies have evaluated the effect of these combined amendments on endemic plant growth. Endemic plants growing on contaminated soils showed higher tolerance toward pollutants compared to plants coming from unpolluted areas. Therefore, the aim of the present study was to evaluate both the effect of amendments (single or combined) on the physicochemical properties of a former mining technosol, and the growth and metal(loid) accumulation ability of endemic Agrostis capillaris plants. This study revealed an improvement in the soil physicochemical properties following the application of amendments, with combined amendments showing better results than the application of just one. On top of this, Agrostis plants performed better on the amended technosols, especially the ones receiving manure, due to its high nutrient content. Finally, based on soil properties, plant growth and the metal(loid) accumulation profile, the use of biochar combined with manure seems to be the most appropriate treatment. Indeed, this treatment showed an improvement in both soil fertility and plant growth. Moreover, Agrostis plants grown in these conditions were among those showing higher root metal(loid) concentration associated with a lower translocation toward aerial parts.

Effect of biochar and redmud amendment combinations on Salix triandra growth, metal(loid) accumulation and oxidative stress response.

Remediation of metal(loid) polluted soils is an important area of research nowadays. In particular, one remediation technique is much studied, phytomanagement. Phytomanagement combines amendment application and plant growth in order to reduce the risk posed by contaminants. Salicaceae plants showed tolerance towards metal(loid)s and the ability to accumulate high amounts of metal(loid)s in their tissue. Amendments are often applied to counterbalance the reduced soil fertility and high metal(loid) concentrations. Two amendments gathered attention over the last decades, biochar (product of biomass pyrolysis), which can be activated for better effects, and redmud (by-product of alumina production). Those two amendments showed ability to improve soil conditions and thus plant growth, although few studied their combined application. Moreover, since metal(loid)s are known to induce the overproduction of reactive oxygen species, it is important to measure the level of oxidative stress in the plant, to which plants respond using enzymatic and non-enzymatic systems. But no studies evaluate the response of Salicaceae plants to metal(loid) stress and amendment application at the biochemical level in a real soil condition. Therefore, a mesocosm study was set up to evaluate the effect of amending a mine soil with redmud combined to diverse biochars on the soil properties and Salix triandra growth, metal(loid) accumulation and stress marker levels. Results showed that all amendment combinations improved the soil fertility, reduced metal(loid) mobility and thus ameliorated Salix triandra growth, which accumulated metal(loid)s mainly in its roots. Moreover, among the different amendment combinations, Salix triandra plants still suffered from oxidative stress when grown on PG soil amended with redmud and chemical activated carbon, showing elevated levels of phenolic compounds and salicinoids and important antioxidant and enzymatic activities. Finally, one treatment showed levels of these stress markers similar or lower than the control, the combination of redmud with steam activated carbon. In conclusion, this treatment seemed a good solution in a phytomanagement strategy using Salix triandra, improving soil conditions and plant growth and reducing oxidative stress level in the plant roots.

Application of biochars and solid fraction of digestate to decrease soil solution Cd, Pb and Zn concentrations in contaminated sandy soils.

Biochar prepared from waste biomass was evaluated as a soil amendment to immobilize metals in two contaminated soils. A 60-day incubation experiment was set up on a French technosol which was heavily contaminated with Pb due to former mining activities. Grass biochar, cow manure biochar (CMB) and two lightwood biochars differing in particle size distribution (LWB1 and LWB2) were amended to the soil at a rate of 2% (by mass). Rhizon soil moisture samplers were employed to assess the Pb concentrations in the soil solution at regular times. After 30 days of incubation, soil solution concentrations in the CMB-amended soil decreased by more than 99% compared to the control. CMB was also applied to a moderately contaminated Flemish soil and resulted in lowered soil solution Cd and Zn concentrations. While the application of 4% CMB resulted in 90% and 80% reductions in soil solution concentrations of Cd and Zn, respectively, the solid fraction of digestate (as a reference) reduced the soil pore water concentrations by only 63% for Cd and 73% for Zn, compared to the concentrations in the control. These results emphasize the potential of biochar to immobilize metals in soil and water systems, thus reducing their phytotoxicity.

Mobility of Pb, Zn, Ba, As and Cd toward soil pore water and plants (willow and ryegrass) from a mine soil amended with biochar.

Mine soils often contain metal(loid)s that may lead to serious environmental problems. Phytoremediation, consisting in covering the soil with specific plants with the possible addition of amendments, represents an interesting way of enhancing the quality of mine soils by retaining contaminants and reducing soil erosion. In order to study the effect of an assisted phytoremediation (with willow and ryegrass) on the properties of soil pore water (SPW), we investigated the impact of amendment with biochar (BC) combined with the planting of willow and ryegrass on the behavior of several metal(loid)s (Pb, Zn, Ba, As, and Cd) in a mine soil. Data on the physicochemical parameters and concentrations of the different metal(loid)s in both SPW and in plant tissues of willow and ryegrass highlight the importance of BC for SPW properties in terms of reductions in soluble concentrations of Pb and Zn, although there was no effect on the behavior of As and Cd. BC also increased soluble concentrations of Ba, probably related to ion release by the BC. By improving major ions available in mine soil, BC improved the lifetime of plants and enhanced their growth. Plant development did not appear to significantly affect the physicochemical parameters of SPW. Willow and ryegrass growing on soil with BC incorporated Cd and Ba into their tissues. The influence of plants on the behavior of metal(loid)s was noticeable only for ryegrass growing in soil with 2% BC, where it modified the behavior of Pb and Ba.

Early molecular events involved in Pinus pinaster Ait. somatic embryo development under reduced water availability: transcriptomic and proteomic analyses.

Maritime pine somatic embryos (SEs) require a reduction in water availability (high gellan gum concentration in the maturation medium) to reach the cotyledonary stage. This key switch, reported specifically for pine species, is not yet well understood. To facilitate the use of somatic embryogenesis for mass propagation of conifers, we need a better understanding of embryo development. Comparison of both transcriptome (Illumina RNA sequencing) and proteome [two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis with mass spectrometry (MS) identification] of immature SEs, cultured on either high (9G) or low (4G) gellan gum concentration, was performed, together with analysis of water content, fresh and dry mass, endogenous abscisic acid (ABA; gas chromatography-MS), soluble sugars (high-pressure liquid chromatography), starch and confocal laser microscope observations. This multiscale, integrated analysis was used to unravel early molecular and physiological events involved in SE development. Under unfavorable conditions (4G), the glycolytic pathway was enhanced, possibly in relation to cell proliferation that may be antagonistic to SE development. Under favorable conditions (9G), SEs adapted to culture constraint by activating specific protective pathways, and ABA-mediated molecular and physiological responses promoting embryo development. Our results suggest that on 9G, germin-like protein and ubiquitin-protein ligase could be used as predictive markers of SE development, whereas protein phosphatase 2C could be a biomarker for culture adaptive responses. This is the first characterization of early molecular mechanisms involved in the development of pine SEs following an increase in gellan gum concentration in the maturation medium, and it is also the first report on somatic embryogenesis in conifers combining transcriptomic and proteomic datasets.

Exploring Phytoremediation Potential: A Comprehensive Study of Flora Inventory and Soil Heavy Metal Contents in the Northeastern Mining Districts of Morocco.

Mining activities produce waste materials and effluents with very high metal concentrations that can negatively impact ecosystems and human health. Consequently, data on soil and plant metal levels are crucial for evaluating pollution severity and formulating soil reclamation strategies, such as phytoremediation. Our research focused on soils and vegetation of a highly contaminated site with potentially toxic metals (Pb, Zn, and Cu) in the Touissit mining districts of eastern Morocco. Vegetation inventory was carried out in three mine tailings of the Touissit mine fields using the "field tower" technique. Here, 91 species belonging to 23 families were inventoried: the most represented families were Poaceae and Asteraceae, and the biological spectrum indicated a predominance of Therophytes (55.12%). From the studied areas, 15 species were selected and collected in triplicate on the tailings and sampled with their corresponding rhizospheric soils, and analyzed for Pb, Zn, and Cu concentrations. Reseda lutea, lotus marocanus, and lotus corniculatus can be considered as hyperaccumulators of Pb, as these plants accumulated more than 1000 mg·kg-1 in their aerial parts. According to TF, these plant species could serve as effective plants for Pb phytoextraction.

Dynamic performance of combined biochar from co-pyrolysis of pig manure with invasive weed: Effect of natural aging on Pb and As mobilization in polluted mining soil.

Due to the natural biochar aging, the improvement of soil quality and immobilization of soil pollutants achieved by biochar may change; understanding the dynamic evolution of the in situ performance of biochar in these roles is essential to discuss the long-term sustainability of biochar remediation. Therefore, in this study, combined biochar from co-pyrolysis of pig manure and invasive Japanese knotweed - P1J1, as well as pure pig manure - PM - and pure Japanese knotweed - JK - derived biochar were applied to investigate their remediation performance in a high As- and Pb-polluted soil with prolonged incubation periods (up to 360 days). Biochar application, especially P1J1 and PM, initially promoted soil pH, dissolved organic carbon, and EC, but the improvements were not constant through time. The JK-treated soil exhibited the highest increase of soil organic matter (OM), followed by P1J1 and then PM, and OM did not change with aging. Biochar, especially P1J1, was a comprehensive nutrient source of Ca, K, Mg, and P to improve soil fertility. However, while soluble cationic Ca, K, and Mg increased with time, anionic P decreased over time, indicating that continuous P availability might not be guaranteed with the aging process. The total microorganism content declined with time; adding biochars slowed down this tendency, which was more remarkable at the later incubation stage. Biochar significantly impeded soil Pb mobility but mobilized soil As, especially in PM- and P1J1-treated soils. However, mobilized As gradually re-fixed in the long run; meanwhile, the excellent Pb immobilization achieved by biochars was slightly reduced with time. The findings of this study offer fresh insights into the alterations in metal(loid)s mobility over an extended duration, suggesting that the potential mobilization risk of As is reduced while Pb mobility slightly increases over time.

Characterization of histidine-aspartate kinase HK1 and identification of histidine phosphotransfer proteins as potential partners in a Populus multistep phosphorelay.

In poplar, we identified proteins homologous to yeast proteins involved in osmosensing multistep phosphorelay Sln1p-Ypd1p-Ssk1p. This finding led us to speculate that Populus cells could sense osmotic stress by a similar mechanism. This study focuses on first and second protagonists of this possible pathway: a histidine-aspartate kinase (HK1), putative osmosensor and histidine phosphotransfer proteins (HPt1 to 10), potential partners of this HK. Characterization of HK1 showed its ability to homodimerize in two-hybrid tests and to act as an osmosensor with a kinase activity in yeast, by functional complementation of sln1Δ sho1Δ strain. Moreover, in plant cells, plasma membrane localization of HK1 is shown. Further analysis on HPts allowed us to isolate seven new cDNAs, leading to a total of 10 different HPts identified in poplar. Interaction tests showed that almost all HPts can interact with HK1, but two of them exhibit stronger interactions, suggesting a preferential partnership in poplar. The importance of the phosphorylation status in these interactions has been investigated with two-hybrid tests carried out with mutated HK1 forms. Finally, in planta co-expression analysis of genes encoding these potential partners revealed that only three HPts are co-expressed with HK1 in different poplar organs. This result reinforces the hypothesis of a partnership between HK1 and these three preferential HPts in planta. Taken together, these results shed some light on proteins partnerships that could be involved in the osmosensing pathway in Populus.

Metal behavior and soil quality changes induced by the application of tailor-made combined biochar: An investigation at pore water scale.

The remediation performance of biochar varies based on the biomass used for its production. Further innovation involves developing tailor-made biochar by combining different raw materials to compensate for the limitations of pure biochar. Therefore, tailor-made combined biochar produced from the co-pyrolysis of pig manure and invasive Japanese knotweed (P1J1), as well as biochars produced from these feedstocks separately, i.e., pure pig manure (PM) and pure Japanese knotweed (JK), were applied to Pb and As contaminated soil to evaluate the biochar-induced changes on soil properties, microbial activity, DOM, and metal and metalloids solubility at the soil pore water scale. Biochar application reduced soluble Pb, whereas enhanced the As mobility; the increased soil pH after biochar addition played a fundamental role in reducing the Pb solubility, as revealed by their significant negative correlation (r = -0.990, p < 0.01). In contrast, the release of dissolved P strongly influenced As mobilization (r = 0.949, p < 0.01), especially in P-rich PM and P1J1 treatments, while JK showed a marginal effect in mobilizing As. Soils treated with PM, P1J1, and JK mainly increased Gram-negative bacteria by 56 %, 52 %, and 50 %, respectively, compared to the control. Fluorescence excitation-emission matrix spectroscopy combined with parallel factor analysis identified three components in pore water DOM, C1 (long wavelength humic-like), C2 (short wavelength humic-like), and C3 (protein-like), which were dominant respectively in the P1J1, JK, and PM-added soil. A principal component analysis (PCA) confirmed that the PM and P1J1 had similar performance and were more associated with releasing P and Mg and specific DOM components (C1 and C3). Meanwhile, P1J1 supplemented soil OM/OC and K, similar to JK. The results of this study suggest that combined biochar P1J1 can comprehensively enhance soil quality, embodying the advantages of pure PM and JK biochar while overcoming their shortcomings.

Importance of Application Rates of Compost and Biochar on Soil Metal(Loid) Immobilization and Plant Growth.

In this study, we investigated the effect of different rates of compost (20%, 40%, 60% w/w) in combination with biochar (0%, 2%, 6% w/w) on soil physiochemical properties and the mobility of arsenic (As) and lead (Pb), in addition to the ability of Arabidopsis thaliana (ecotype Columbia-0) to grow and accumulate metal(loid)s. All modalities improved pH and electrical conductivity, stabilized Pb and mobilized As, but only the mixture of 20% compost and 6% biochar improved plant growth. Plants in all modalities showed a significant reduction in root and shoot Pb concentrations compared to the non-amended technosol. In contrast, As shoot concentration was significantly lower for plants in all modalities (except with 20% compost only) compared to non-amended technosol. For root As, plants in all modalities showed a significant reduction except for the mixture of 20% compost and 6% biochar. Overall, our results indicate that the mixture of 20% compost with 6% biochar emerged as the optimum combination for improving plant growth and As uptake, making it the possible optimum combination for enhancing the efficiency of land reclamation strategies. These findings provide a foundation for further research on the long-term effects and potential applications of the compost-biochar combination in improving soil quality.

Different species of Bradyrhizobium from symbiovars genistearum and retamae nodulate the endemic Retama dasycarpa in the High Atlas Mountains.

Retama dasycarpa is an endemic Retama species native to the cold semi-arid bioclimates of the High Atlas Mountains in Morocco. In this work, we analyzed the diversity of the microsymbionts nodulating this plant and their different phenotypic and symbiotic characteristics. Phylogenetic analysis of the 16S rRNA gene revealed that the tested isolates clustered in the Bradyrhizobium genus. Multilocus sequence analyses of four housekeeping genes (recA, gyrB, glnII and atpD) for 12 selected strains grouped them into four clusters close to B. lupini USDA 3051T, B. frederickii CNPSo 3446T, B. valentinum LmjM3T and B. retamae Ro19T. The individual phylogenies of these core genes and the symbiotic genes nodC, nodA and nifH were congruent. These isolates showed a broad host range, being able to nodulate different legume hosts, such as R. sphaerocarpa, R. monosperma, Lupinus luteus, Cytisus grandiflorus andChamaecytisus albidus, but not Phaseolus vulgaris or Glycine max. They all had a similar metabolic capacity, using the majority of the carbohydrates and amino acids tested as sole sources of carbon and nitrogen. Furthermore, out of the 12 selected strains, some displayed plant growth-promoting features, with six of them solubilizing phosphate and three of them producing siderophores. The present work provides, for the first time, a detailed description about the microsymbionts associated with the endemic legume R. dasycarpa.

Identification of five B-type response regulators as members of a multistep phosphorelay system interacting with histidine-containing phosphotransfer partners of Populus osmosensor.

BACKGROUND: In plants, the multistep phosphorelay signaling pathway mediates responses to environmental factors and plant hormones. This system is composed of three successive partners: hybrid Histidine-aspartate Kinases (HKs), Histidine-containing Phosphotransfer proteins (HPts), and Response Regulators (RRs). Among the third partners, B-type RR family members are the final output elements of the pathway; they act as transcription factors and clearly play a pivotal role in the early response to cytokinin in Arabidopsis. While interactions studies between partners belonging to the multistep phosphorelay system are mainly focused on protagonists involved in cytokinin or ethylene pathways, very few reports are available concerning partners of osmotic stress signaling pathway. RESULTS: In Populus, we identified eight B-type RR proteins, RR12-16, 19, 21 and 22 in the Dorskamp genotype. To assess HPt/B-type RR interactions and consequently determine potential third partners in the osmosensing multistep phosphorelay system, we performed global yeast two-hybrid (Y2H) assays in combination with Bimolecular Fluorescence Complementation (BiFC) assays in plant cells. We found that all B-type RRs are able to interact with HPt predominant partners (HPt2, 7 and 9) of HK1, which is putatively involved in the osmosensing pathway. However, different profiles of interaction are observed depending on the studied HPt. HPt/RR interactions displayed a nuclear localization, while the nuclear and cytosolic localization of HPt and nuclear localization of RR proteins were validated. Although the nuclear localization of HPt/RR interaction was expected, this work constitutes the first evidence of such an interaction in plants. Furthermore, the pertinence of this partnership is reinforced by highlighting a co-expression of B-type RR transcripts and the other partners (HK1 and HPts) belonging to a potential osmosensing pathway. CONCLUSION: Based on the interaction studies between identified B-type RR and HPt proteins, and the co-expression analysis of transcripts of these potential partners in poplar organs, our results favor the model that RR12, 13, 14, 16 and 19 are able to interact with the main partners of HK1, HPt2, 7 and 9, and this HPt/RR interaction occurs within the nucleus. On the whole, the five B-type RRs of interest could be third protagonists putatively involved in the osmosensing signaling pathway in Populus.