The composition of root exudates is modulated by several environmental factors, and it remains unclear how that affects beneficial rhizosphere or inoculated microorganisms under heavy metal (HM) contamination. Therefore, we evaluated the transcriptional response of Pseudomonas putida E36 (a Miscanthus x giganteus isolate with plant growth promotion-related properties) to Cd, Pb and Zn in an in vitro study implementing root exudates from M. x giganteus. To collect root exudates and analyse their composition plants were grown in a pot experiment under HM and control conditions. Our results indicated higher exudation rate for plants challenged with HM. Further, out of 29 organic acids identified and quantified in the root exudates, 8 of them were significantly influenced by HM (e.g., salicylic and terephthalic acid). The transcriptional response of P. putida E36 was significantly affected by the HM addition to the growth medium, increasing the expression of several efflux pumps and stress response-related functional units. The additional supplementation of the growth medium with root exudates from HM-challenged plants resulted in a downregulation of 29% of the functional units upregulated in P. putida E36 as a result of HM addition to the growth medium. Surprisingly, root exudates + HM downregulated the expression of P. putida E36 functional units related to plant colonization (e.g., chemotaxis, motility, biofilm formation) but upregulated its antibiotic and biocide resistance compared to the control treatment without HM. Our findings suggest that HM-induced changes in root exudation pattern may attract beneficial bacteria that are in turn awarded with organic nutrients, helping them cope with HM stress. However, it might affect the ability of these bacteria to colonize plants growing in HM polluted areas. Those findings may offer an insight for future in vivo studies contributing to improvements in phytoremediation measures.

Rhizospheric microorganisms such as denitrifying bacteria are able to affect ‘rhizobioaugmention’ in aquatic plants and can help boost wastewater purification by benefiting plant growth, but little is known about their effects on the production of plant root exudates, and how such exudates may affect microorganismal nitrogen removal. Here, we assess the effects of the rhizospheric Pseudomonas inoculant strain RWX31 on the root exudate profile of the duckweed Spirodela polyrrhiza, using gas chromatography/mass spectrometry. Compared to untreated plants, inoculation with RWX31 specifically induced the exudation of two sterols, stigmasterol and β-sitosterol. An authentic standard assay revealed that stigmasterol significantly promoted nitrogen removal and biofilm formation by the denitrifying bacterial strain RWX31, whereas β-sitosterol had no effect. Assays for denitrifying enzyme activity were conducted to show that stigmasterol stimulated nitrogen removal by targeting nitrite reductase in bacteria. Enhanced N removal from water by stigmasterol, and a synergistic stimulatory effect with RWX31, was observed in open duckweed cultivation systems. We suggest that this is linked to a modulation of community composition of nirS- and nirK-type denitrifying bacteria in the rhizosphere, with a higher abundance of Bosea, Rhizobium, and Brucella, and a lower abundance of Rubrivivax. Our findings provide important new insights into the interaction of duckweed with the rhizospheric bacterial strain RWX31 and their involvement in the aquatic N cycle and offer a new path toward more effective bio-formulations for the purification of N-polluted waters.

Heavy metal stress in soil accelerates the plant root exudation of organic ligands. The degradation of exudate ligands can be fundamental to controlling the complexation of heavy metals. However, this process remains poorly understood. Here, we investigated the relationship between the transformation of glycine, a representative amino acid exudate, and cadmium/lead mobility in soils. Two 48-h incubation experiments were conducted after glycine addition to the soils. Parameters related to glycine distribution and degradation, Cd/Pb mobility, and the formation of glycine-Cd complex were analyzed. Glycine addition gradually decreased the Cd and Pb mobility throughout the 48-h incubation. By the end of the experiment, the CaCl₂-extracted Cd and Pb concentrations decreased by 63.5% and 43.6%, respectively. The glycine mineralization was strong in the first 6 h, as indicated by a sharp decrease in CO₂ efflux rates from 10.04 ± 0.62 to 3.51 ± 0.07 mg C–CO₂ kg⁻¹ soil h⁻¹. The mineralization rates notably decreased after 6 h. The comparisons of dissolved organic carbon and hydrolyzable amino acid contents indicated that glycine mineralization in solution (95.6%) was much stronger than that in soil solids (49.3%). At the end of incubation, 0.22 mmol kg⁻¹ glycine remained in soil solids. The remaining glycine provided sufficient sorption sites for Cd²⁺ and Pb²⁺, resulting in enhanced metal fixation via complexation. Comparisons of zeta potentials supported the formation of the glycine-Cd complex. The Cd and Pb immobilization processes could be attributed to metal-glycine complex formation, sorption re-equilibrium, and glycine degradation. These findings emphasize that the biogeochemical processes of glycine, derived from root exudates or protein degradation products, increased the sorption of heavy metals to soils and thus reduced their toxicity to plants.

Due to relatively high chelant dosages and potential environmental risks it is necessary to explore different approaches in the remediation of metal-contaminated soils. The present study focussed on the removal of metals (As, Cd, Cu, Pb and Zn) from a multiple metal-contaminated soil by growing Brassica carinata plants in succession to spontaneous metallicolous populations of Pinus pinaster, Plantago lanceolata and Silene paradoxa. The results showed that the growth of the metallicolous populations increased the extractable metal levels in the soil, which resulted in a higher accumulation of metals in the above-ground parts of B. carinata. Root exudates of the three metallicolous species were analysed to elucidate their possible role in the enhanced metal availability. The presence of metals stimulated the exudation of organic and phenolic acids as well as flavonoids. It was suggested that root exudates played an important role in solubilising metals in soil and in favouring their uptake by roots.

Restoring enzyme function in barren, brownfield soils using green strategies can improve microbial functioning and enable phytoremediation. It is known that adding simple, readily metabolized substrates secreted by growing plant roots (root exudates) or a laboratory prepared solution of root exudates (artificial root exudates) can stimulate soil microbial function. It is not known whether and how well this strategy works in a contaminated, low functioning soil from an industrial barren site because contaminants in the barren soil might inhibit microbial survival and functioning, or the microbial community might not be adapted to functionally benefit from root exudates. The objective of this study was to determine whether artificial root exudates stimulate microbial function in a barren soil. We collected soils from a barren brownfield (25R) site and an adjacent vegetated brownfield site (25F), with low and high enzyme activities, respectively. We subjected both soils to three treatments: switchgrass (native to the site), artificial root exudates, and a combination of switchgrass and artificial root exudates. We measured enzymatic activity, plant growth, soil moisture, organic matter content, and easily extractable glomalin content over 205 days. By day 157, artificial root exudates increased the phosphatase activity by 9-fold in previously vegetated brownfield soil and by 351-fold in barren brownfield soil. When exudates were added to the barren soil, the plant shoot mass was higher (52.2 ± 2.5 mg) than when they were not (35.4 ± 3.6 mg). In both soils, adding artificial root exudates significantly increased the percent moisture, organic matter, and glomalin content. Treating contaminated, barren soil with artificial root exudates resulted in increased soil microbial function and improved soil properties that might promote a hospitable habitat to support vegetation in such extreme environments. Summary: We added artificial root exudates to stimulate enzymatic function in two contaminated soils. Plant shoot mass, soil percent moisture, glomalin content, and organic matter content significantly increased due to the addition of artificial root exudates to the study soils. Microbially-mediated phosphatase activity was established in a barren, previously inactive, polluted soil.

Soil heavy metal contamination has increasingly become a serious environmental issue globally, nearing crisis proportions. There is an urgent need to find environmentally friendly materials to remediate heavy-metal contaminated soils. With the continuing maturation of research on using biochar (BC) for the remediation of contaminated soil, nano-biochar (nano-BC), which is an important fraction of BC, has gradually attracted increasing attention. Compared with BC, nano-BC has unique and useful properties for soil remediation, including a high specific surface area and hydrodynamic dispersivity. The efficacy of nano-BC for immobilization of non-degradable heavy-metal contaminants in soil systems, however, is strongly affected by plant rhizosphere processes, and there is very little known about the role that nano-BC play in these processes. The rhizosphere represents a dynamically complex soil environment, which, although having a small thickness, drives potentially large materials fluxes into and out of plants, notably agricultural foodstuffs, via large diffusive gradients. This article provides a critical review of over 140 peer-reviewed papers regarding nano-BC-rhizosphere interactions and the implications for the remediation of heavy-metal contaminated soils. We conclude that, when using nano-BC to remediate heavy metal-contaminated soil, the relationship between nano-BC and rhizosphere needs to be considered. Moreover, the challenges to extending our knowledge regarding the environmental risk of using nano-BC for remediation, as well as further research needs, are identified.

Dioecious plants show sexual differences in resistance traits to abiotic stresses. However, the effects of exogenous pesticide application on female and male plant growth and their associated adaptation mechanisms are unclear. Our study investigated the effects of the broad-spectrum pesticide lambda-cyhalothrin (λ-CY) on dioecious Populus cathayana growth and explored the factors through which λ-CY changed the rhizosphere bacterial community and physicochemical soil properties via sex-specific metabolomics. The sequential application of λ-CY significantly suppressed male shoot- and root biomass, with little effect on the growth of females. Females possessed a higher intrinsic chemo-diversity within their root exudates, and their levels of various metabolites (sugars, fatty acids, and small organic acids) increased after exposure to λ-CY with consequences on bacterial community composition. Maintaining high bacterial alpha diversity and recruiting specific bacterial groups slowed down the loss of rhizosphere nutrients in females. In contrast, the reduction in bacterial alpha diversity and network structure stability in males was associated with lower rhizosphere nutrient availability. Spearman's correlation analysis revealed that several bacterial groups were positively correlated with the root secretion of lipids and organic acids, suggesting that these metabolites can affect the soil bacterial groups actively involved in the nutrient pool. This study provided novel insights that root exudates and soil microbial interactions may mediate sex-specific differences in response to pesticide application.

Straw amendment and plant root exudates modify the quality and quantities of soil dissolved organic matter (DOM) and then manipulate the fractions of soil selenium (Se) and its bioavailability. Two typical soils with distinct pH were selected to investigate the effect of different contributors on DOM-Se in soil. The mechanisms relying on the variation in DOM characteristics (quality, quantity and composition) were explored by UV–Vis, ATR-FTIR and 3D-EEM. Straw amendment significantly (p < 0.05) suppressed the selenate bioavailability. The reduction in wheat Se content was greater in krasnozems than in Lou soil, as more HA fraction appeared in krasnozems. The root exudates of wheat mainly elevated the low molecular hydrophilic compounds (Hy) in soil, which contributed to the SOL-Hy-Se fractions and thus grain Se in soils (p < 0.01). However, straw amendment promoted DOM transforming from small molecules (Hy and FA) to aromatic large molecules (HA), when accompanied with the reduction and retention of Se associated with these molecules. As a result, selenium bioavailability and toxicity reduced with DOM amendment and DOM-Se transformation.

Microbial methylmercury (MeHg) production in contaminated soil-rice systems and its accumulation in rice pose health risks to consumers, especially those in Asia. However, the mechanism responsible for microbial MeHg production in paddy soils is far from clear. While previous studies examined the effect of soil and microbial factors on soil MeHg levels, in this work we explored the impact of rice cultivation itself on microbial MeHg production, focusing on the root exudate organic matter as a potential source of electron donors for microbial methylators. Effects of the cultivation of two rice cultivars, Heigu246 (H-rice) and Neiwuyou8015 (N-rice), on MeHg production in soils were therefore investigated in pot and batch incubation experiments. Soil MeHg levels measured in H-rice treatment during the heading and harvest stages were 18–49% higher than in the control and 23–108% higher than in N-rice treatment. Consequently, MeHg levels in grain, straw, and root were 38%, 81%, and 40% higher in H-rice than those in N-rice, which was mainly attributed to cultivar-specific MeHg production in soils. Results of the batch experiments suggested that root exudate organic matter could be responsible for MeHg production in soils during rice cultivation, by increasing the abundances of potential microbial methylators. For instance, root exudate organic matter increased copy numbers of Hg methylation genes (hgcA) in soils 4.1-fold. Furthermore, the 211% higher concentration of acetate (a key electron donor for microbial methylators) in the root exudate of H-rice could account for the higher MeHg production under H-rice than N-rice cultivation. Our results suggest that root exudate organic matter, especially acetate, as its key component, contributes to the elevated soil MeHg concentrations during rice cultivation. The proposed mechanism provides new insights into the elevated risk of MeHg production in contaminated soil-rice systems, as well as cultivar-specific MeHg bioaccumulation.

Understanding the phytoavailability of bound residues of polycyclic aromatic hydrocarbons (PAHs) in soils is essential to assessing their environmental fate and risks. This study investigated the release and plant uptake of bound PAH residues (reference to parent compounds) in field contaminated soils after the removal of extractable PAH fractions. Plant pot experiments were performed in a greenhouse using ryegrass (Lolium multiflorum Lam.) to examine the phytoavailablility of bound PAH residues, and microcosm incubation experiments with and without the addition of artificial root exudates (AREs) or oxalic acid were conducted to examine the effect of root exudates on the release of bound PAH residues. PAH accumulation in the ryegrass after a 50-day growth period indicated that bound PAH residues were significantly phytoavailable. The extractable fractions, including the desorbing and non-desorbing fractions, dominated the total PAH concentrations in vegetated soils after 50 days, indicating the transfer of bound PAH residues to the extractable fractions. This transfer was facilitated by root exudates. The addition of AREs and oxalic acid to test soils enhanced the release of bound PAH residues into their extractable fractions, resulting in enhanced phytoavailability of bound PAH residues in soils. This study provided important information regarding environmental fate and risks of bound PAH residues in soils.