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.

The symbiosis of corals, zooxanthellae, and microbes is the foundation of the coral reef ecosystem. In addition to global warming, marine pollutants are another important factor causing the breakdown of coral symbiosis. Benzo(a)pyrene (BaP) is a globally widespread marine environmental pollutant that poses a severe threat to marine ecosystems. However, responses of coral symbionts to global marine pollutant stress remain unclear. In this study, we selected Acropora formosa as the target coral to explore its response to 50 μg L⁻¹ BaP stress using diaPASEF proteomics and 16s rRNA microbiome analysis. The results showed that: 1) the coral symbionts were sensitive to BaP stress; 2) the photosynthetic system of zooxanthellae was crucial for the balance of symbiotic relationships; 3) the destruction of the photosynthetic system induced a zooxanthellae hypoxic stress response; 4) corals adapted to BaP stress by promoting non-essential protein degradation and changing energy metabolism strategies; 5) symbiotic bacteria showed strong adaptability to BaP. This study not only fills the gap in understanding the response mechanism of coral symbionts under BaP stress, but also provides fundamental data for coral reef protection strategies.

DNA-based analyses of bacterial communities were performed to identify the bacteria co-occurring with cyanobacterial blooms in samples collected at a single site over 2 years. Microcystis aeruginosa was the most predominant species (81% in 2018, and 94% in 2019) within the phylum Cyanobacteria, and microcystins were detected during all cyanobacterial blooms. The stereo microscope and scanning electron microscope observations showed bacterial associations on and around the aggregated M. aeruginosa cells. Culture-independent analyses of filtered bacterial communities showed that the Flavobacterium species in phylum Bacteroidetes (19%) was dominant in the cyanobacterial phycosphere, followed by the Limnohabitans species in Betaproteobacteria (11%). Using principal component analysis, major bacterial genus, including Microcystis and Flavobacterium species, were clustered during cyanobacterial blooms in both years. To identify key bacterial species that develop long-term symbiosis with M. aeruginosa, another culture-independent analysis was performed after the environmental sample had been serially subcultured for 1 year. Interestingly, Brevundimonas (14%) was the most dominant species, followed by Porphyrobacter (7%) and Rhodobacter (3.5%) within the Alphaproteobacteria. Screening of 100 colonies from cyanobacterial bloom samples revealed that the majority of culturable bacteria belonged to Gammaproteobacteria (28%) and Betaproteobacteria (57%), including Pseudomonas, Curvibacter, and Paucibacter species. Several isolates of Brevundimonas, Curvibacter, and Pseudomonas species could promote the growth of axenic M. aeruginosa PCC7806. The sensitivity of M. aeruginosa PCC7806 cells to different environmental conditions was monitored in bacteria-free pristine freshwater, indicating that nitrogen addition promotes the growth of M. aeruginosa.

In this study, coral soft tissue, skeleton and zooxanthellae, as well as their ambient sediment and seawater were analyzed for polycyclic aromatic hydrocarbons (PAHs) with a special focus on perylene. Samples were collected from two different environments: the Kharg Island, which is affected by numerous anthropogenic stressors and Larak Island, which is mainly used for recreational and fishing activities and is characterized by dense vegetation. The heaviest loadings of PAHs were observed on Kharg Island, yet higher concentrations of perylene were detected on Larak Island and it was identified as the prevailing compound in this area. Pyrogenic perylene sources were prevailing on Kharg Island, whereas the perylene on Larak Island was determined to be of natural origin. After analyzing the biological samples, higher perylene concentrations were observed in zooxanthellae than in tissue and skeleton. The lowest and the highest perylene loadings were found in the tissue and skeleton of Platygyra daedalea and Porites lutea, respectively. This applies to both reefs. We found that perylene distribution in the corals and their ambient environment follows an irregular pattern, demonstrating remarkable effects from the local inputs. The lipid content in the coral tissue and the location of the coral colony were deduced to be the main factors affecting perylene distribution in corals. On Larak Island, a significant correlation between perylene loadings in sediment and corals was observed. On Kharg Island, a strong interaction between the water column and the corals was detected. The symbiotic relationship between the corals and zooxanthellae might play the most significant role in bioconcentration and bioaccumulation of perylene. Due to the insolubility of PAHs, they could be transferred through a food chain to zooxanthellae and eventually deposited in the coral bodies.

Very few studies reported the potential of arbuscular mycorrhizal symbiosis to dissipate hydrocarbons in aged polluted soils. The present work aims to study the efficiency of arbuscular mycorrhizal colonized wheat plants in the dissipation of alkanes and polycyclic aromatic hydrocarbons (PAHs). Our results demonstrated that the inoculation of wheat with Rhizophagus irregularis allowed a better dissipation of PAHs and alkanes after 16 weeks of culture by comparison to non-inoculated condition. These dissipations observed in the inoculated soil resulted from several processes: (i) a light adsorption on roots (0.5% for PAHs), (ii) a bioaccumulation in roots (5.7% for PAHs and 6.6% for alkanes), (iii) a transfer in shoots (0.4 for PAHs and 0.5% for alkanes) and mainly a biodegradation. Whereas PAHs and alkanes degradation rates were respectively estimated to 12 and 47% with non-inoculated wheat, their degradation rates reached 18 and 48% with inoculated wheat. The mycorrhizal inoculation induced an increase of Gram-positive and Gram-negative bacteria by 56 and 37% compared to the non-inoculated wheat. Moreover, an increase of peroxidase activity was assessed in mycorrhizal roots. Taken together, our findings suggested that mycorrhization led to a better hydrocarbon biodegradation in the aged-contaminated soil thanks to a stimulation of telluric bacteria and hydrocarbon metabolization in mycorrhizal roots.