Mercury pollution is an issue of global concern. In Colombia, the use of contaminated water for food crop irrigation and artisanal mining contributes to mercury pollution in soil, affecting food production and restoration of disturbed areas. Mycorrhizal fungi are symbionts that provide benefits to plants including resistance to heavy metals, but fungal effects on germination remain to be fully described. This study tested the effect of mercury and mycorrhizal fungi on Lactuca sativa seed germination. A 2x5 completely randomized factorial experiment was developed to assess the effect of five HgCl2 polluted treatments, two mycorrhizal treatments (i.e., with inoculum, without inoculum), and the interaction of both factors on seed germination, seedling root colonization, pH, and final water content. In samples with no mercury pollution, mycorrhizal fungi had an inhibitory effect on seed germination. Likewise, the effect of mercury on seed germination is significantly inhibitory. However, pots inoculated with arbuscular mycorrhizal fungi showed constant germination probabilities, independently of mercury concentration. According to the best model determined for the data, a key step in the mitigation of mercury toxicity in seed germination is to prevent substrate pH changes. The environmental conditions of the experiment contributed to densely activate populated biomass of inoculum, which promoted root invasion from various points. Overall, the presence of mycorrhizal fungi in seedbeds could lead to a reduced number of plant individuals. However, the use of fungal inoculum in polluted environments, highly contributes to plant establishment, which is relevant in further vegetable cultivations growing in soil polluted areas.

Phytostabilisation can benefit from phytostimulatory rhizobacteria. Forty-three bacterial strains were isolated from the roots of the metallicolous legume Anthyllis vulneraria ssp. carpatica grown in a highly contaminated mine tailing (total Cd, Pb and Zn were up to 1200; 34,000; and 170,000 mg kg(-1), respectively). We aimed at evaluating their phytostimulatory effects on the development of leguminous metallophytes. Strains were screened for fluorescent siderophores and auxin synthesis, inorganic P solubilisation and 1-amino-cyclopropane-1-carboxylate deaminase (ACCd) activity to define a subset of 11 strains that were inoculated on the leguminous metallophytes A. vulneraria and Lotus corniculatus grown in diluted mine spoil (Zn 34,653; Pb 6842; and Cd 242, all in mg kg-1). All strains were affiliated to Pseudomonas spp. (except two), synthetised auxins and siderophores and solubilised P (except three), and seven of them were ACCd positive. The inoculation effects (shoot-root-nodule biomass, chlorophyll content) depended on legume species and bacterial strain genotype. Phytostimulation scores were unrelated to siderophore/auxin synthesis and P solubilisation rates. Inoculations of the strain nos. 17-43 triggered a 1.2-fold significant increase in the chlorophyll content of A. vulneraria. Chlorophyll content and root biomass of L. corniculatus were significantly increased following the inoculations of the strain nos. 17-22 (1.5-1.4-fold, respectively). The strongest positive effects were related to increases in the nodule biomass of L. corniculatus in the presence of three ACCd-positive strains (1.8-fold), one of which was the highest auxin producer. These data suggest to focus on interactions between ACCd activity and auxin synthesis to enhance nodulation of metallicolous legumes.

Phytostabilisation can benefit from phytostimulatory rhizobacteria. Forty-three bacterial strains were isolated from the roots of the metallicolous legume Anthyllis vulneraria ssp. carpatica grown in a highly contaminated mine tailing (total Cd, Pb and Zn were up to 1200; 34,000; and 170,000 mg kg(-1), respectively). We aimed at evaluating their phytostimulatory effects on the development of leguminous metallophytes. Strains were screened for fluorescent siderophores and auxin synthesis, inorganic P solubilisation and 1-amino-cyclopropane-1-carboxylate deaminase (ACCd) activity to define a subset of 11 strains that were inoculated on the leguminous metallophytes A. vulneraria and Lotus corniculatus grown in diluted mine spoil (Zn 34,653; Pb 6842; and Cd 242, all in mg kg-1). All strains were affiliated to Pseudomonas spp. (except two), synthetised auxins and siderophores and solubilised P (except three), and seven of them were ACCd positive. The inoculation effects (shoot-root-nodule biomass, chlorophyll content) depended on legume species and bacterial strain genotype. Phytostimulation scores were unrelated to siderophore/auxin synthesis and P solubilisation rates. Inoculations of the strain nos. 17-43 triggered a 1.2-fold significant increase in the chlorophyll content of A. vulneraria. Chlorophyll content and root biomass of L. corniculatus were significantly increased following the inoculations of the strain nos. 17-22 (1.5-1.4-fold, respectively). The strongest positive effects were related to increases in the nodule biomass of L. corniculatus in the presence of three ACCd-positive strains (1.8-fold), one of which was the highest auxin producer. These data suggest to focus on interactions between ACCd activity and auxin synthesis to enhance nodulation of metallicolous legumes.

Being signaling molecules, nitric oxide (NO) and hydrogen sulfide (H₂S) can mediate a wide range of physiological processes caused by plant metal toxicity. Moreover, legume-rhizobium symbiosis has gained increasing attention in mitigating heavy metal stress. However, systematic regulatory mechanisms used for the exogenous application of signaling molecules to alter the resistance of legume-rhizobium symbiosis under metal stress are currently unknown. In this study, we examined the exogenous effects of sodium nitroprusside (SNP) as an NO donor additive and sodium hydrosulfide (NaHS) as a H₂S donor additive on the phytotoxicity and soil quality of alfalfa (Medicago sativa)-rhizobium symbiosis in lead/cadmium (Pb/Cd)-contaminated soils. Results showed that rhizobia inoculation markedly promoted alfalfa growth by increasing chlorophyll content, fresh weight, and plant height and biomass. Compared to the inoculated rhizobia treatment alone, the addition of NO and H₂S significantly reduced the bioaccumulation of Pb and Cd in alfalfa-rhizobium symbiosis, respectively, thus avoiding the phytotoxicity caused by the excessive presence of metals. The addition of signaling molecules also alleviated metal-induced phytotoxicity by increasing antioxidant enzyme activity and inhibiting the level of lipid peroxidation and reactive oxygen species (ROS) in legume-rhizobium symbiosis. Also, signaling molecules improved soil nutrient cycling, increased soil enzyme activities, and promoted rhizosphere bacterial community diversity. Both partial least squares path modeling (PLS-PM) and variation partitioning analysis (VPA) identified that using signaling molecules can improve plant growth by regulating major controlling variables (i.e., soil enzymes, soil nutrients, and microbial diversity/plant oxidative damage) in legume-rhizobium symbiosis. This study offers integrated insight that confirms that the exogenous application of signaling molecules can enhance the resistance of legume-rhizobium symbiosis under metal toxicity by regulating the biochemical response of the plant-soil system, thereby minimizing potential health risks.

Microplastics are widespread emerging contaminants that have been found globally in the marine and freshwater ecosystem, but there is limited knowledge regarding its impact on coral reef ecosystem and underpinning mechanism. In the present study, using Pocillopora damicornis as a model, we investigated cytological, physiological, and molecular responses of a scleractinian coral to acute microplastic exposure. No significant changes were observed in the density of symbiotic zooxanthellae during the entire period of microplastic exposure, while its chlorophyll content increased significantly at 12 h of microplastic exposure. We observed significant increases in the activities of antioxidant enzymes such as superoxide dismutase and catalase, significant decrease in the detoxifying enzyme glutathione S-transferase and the immune enzyme alkaline phosphatase, but no change in the other immune enzyme phenoloxidase during the whole experiment period. Transcriptomic analysis revealed 134 significantly up-regulated coral genes at 12 h after the exposure, enriched in 11 GO terms mostly related to stress response, zymogen granule, and JNK signal pathway. Meanwhile, 215 coral genes were significantly down-regulated at 12 h after exposure, enriched in 25 GO terms involved in sterol transport and EGF-ERK1/2 signal pathway. In contrast, only 12 zooxanthella genes exhibited significant up-regulation and 95 genes down-regulation at 12 h after the microplastic exposure; genes regulating synthesis and export of glucose and amino acids were not impacted. These results suggest that acute exposure of microplastics can activate the stress response of the scleractinian coral P. damicornis, and repress its detoxification and immune system through the JNK and ERK signal pathways. These demonstrate that microplastic exposure can compromise the anti-stress capacity and immune system of the scleractinian coral P. damicornis, despite the minimal impact on the abundance and major photosynthate translocation transporters of the symbiont in the short term.

The role of indigenous and non-indigenous arbuscular mycorrhizal fungi (AMF) on As uptake by Plantago lanceolata L. growing on substrate originating from mine waste rich in As was assessed in a pot experiment. P. lanceolata inoculated with AMF had higher shoot and root biomass and lower concentrations of As in roots than the non-inoculated plants. There were significant differences in As concentration and uptake between different AMF isolates. Inoculation with the indigenous isolate resulted in increased transfer of As from roots to shoots; AMF from non-polluted area apparently restricted plants from absorbing As to the tissue; and plants inoculated with an AMF isolate from Zn–Pb waste showed strong As retainment within the roots. Staining with dithizone indicated that AMF might be actively involved in As accumulation. The mycorrhizal colonization affected also the concentration of Cd and Zn in roots and Pb concentration, both in shoots and roots.

Global warming and local disturbances such as pollution cause several impacts on coral reefs. Among them is the breakdown of the symbiosis between host corals and photosynthetic symbionts, which is often a consequence of oxidative stress. Therefore, we investigated if the combined effects of thermal stress and copper (Cu) exposure change the trophic behavior and oxidative status of the reef-building coral Mussismilia harttii. Coral fragments were exposed in a mesocosm system to three temperatures (25.0, 26.6 and 27.3 °C) and three Cu concentrations (2.9, 5.4 and 8.6 μg L⁻¹). Samples were collected after 4 and 12 days of exposure. We then (i) performed fatty acid analysis by gas chromatography-mass spectrometry to quantify changes in stearidonic acid and docosapentaenoic acid (autotrophy markers) and cis-gondoic acid (heterotrophy marker), and (ii) assessed the oxidative status of both host and symbiont through analyses of lipid peroxidation (LPO) and total antioxidant capacity (TAC). Our findings show that trophic behavior was predominantly autotrophic and remained unchanged under individual and combined stressors for both 4- and 12-day experiments; for the latter, however, there was an increase in the heterotrophy marker. Results also show that 4 days was not enough to trigger changes in LPO or TAC for both coral and symbiont. However, the 12-day experiment showed a reduction in symbiont LPO associated with thermal stress alone, and the combination of stressors increased their TAC. For the coral, the isolated effects of increase in Cu and temperature led to an increase in LPO. The effects of combined stressors on trophic behavior and oxidative status were not much different than those from the isolated effects of each stressor. These findings highlight that host and symbionts respond differently to stress and are relevant as they show the physiological response of individual holobiont compartments to both global and local stressors.

Silver (Ag) engineered nanomaterials (ENMs) are being released into waste streams and are being discharged, largely as Ag2S aged-ENMs (a-ENMs), into agroecosystems receiving biosolids amendments. Recent research has demonstrated that biosolids containing an environmentally relevant mixture of ZnO, TiO2, and Ag ENMs and their transformation products, including Ag2S a-ENMs, disrupted the symbiosis between nitrogen-fixing bacteria and legumes. However, this study was unable to unequivocally determine which ENM or combination of ENMs and a-ENMs was responsible for the observed inhibition. Here, we examined further the effects of polyvinylpyrollidone (PVP) coated pristine Ag ENMs (PVP-Ag), Ag2S a-ENMs, and soluble Ag (as AgSO4) at 1, 10, and 100 mg Ag kg−1 on the symbiosis between the legume Medicago truncatula and the nitrogen-fixing bacterium, Sinorhizobium melliloti in biosolids-amended soil. Nodulation frequency, nodule function, glutathione reductase production, and biomass were not significantly affected by any of the Ag treatments, even at 100 mg kg−1, a concentration analogous to a worst-case scenario resulting from long-term, repeated biosolids amendments. Our results provide additional evidence that the disruption of the symbiosis between nitrogen-fixing bacteria and legumes in response to a mixture of ENMs in biosolids-amended soil reported previously may not be attributable to Ag ENMs or their transformation end-products. We anticipate these findings will provide clarity to regulators and industry regarding potential unintended consequences to terrestrial ecosystems resulting from of the use of Ag ENMs in consumer products.

Globally, the legume–rhizobia symbiosis, contained within specialised organs called root nodules, is thought to add at least 30 Tg N annually to agricultural land. The growth and functioning of a modern white clover (Trifolium repens cv. Crusader) and red clover (T. pratense cv. Merviot) cultivar were investigated in current and future ozone scenarios in solardomes. Both cultivars developed leaf injury and had significant reductions in root biomass and root nodule number in response to ozone, with Crusader also displaying a reduced size and mass of nodules. In-situ measurements of N-fixation in Crusader by acetylene reduction assay revealed reduced N-fixation rates in a future scenario with an increased background and moderate peaks of ozone. The implications for the sustainability of temperate pasture are discussed.

The effect of arbuscular mycorrhizal fungi (AMF) on growth and element uptake by Ni-hyperaccumulating plant, Berkheya coddii, was studied. Plants were grown under laboratory conditions on ultramafic soil without or with the AM fungi of different origin. The AM colonization, especially with the indigenous strain, significantly enhanced plants growth and their survival. AMF affected also the elemental concentrations that were studied with Particle-induced X-ray emission (PIXE). AMF (i) increased K and Fe in shoots, Zn and Mn in roots, P and Ca both, in roots and shoots; (ii) decreased Mn in shoots, Co and Ni both, in shoots and roots. Due to higher biomass of mycorrhizal plants, total Ni content was up to 20 times higher in mycorrhizal plants compared to the non-mycorrhizal ones. The AMF enhancement of Ni uptake may therefore provide an improvement of a presently used technique of nickel phytomining.