Root-associated microbiota provide great fitness to hosts under environmental stress. However, the underlying microecological mechanisms controlling the interaction between heavy metal-stressed ...plants and the microbiota are poorly understood. In this study, we screened and isolated representative amplicon sequence variants (strain M4) from rhizosphere soil samples of Trifolium repens L. growing in areas with high concentrations of heavy metals. To investigate the microecological mechanisms by which T. repens adapts to heavy metal stress in abandoned mining areas, we conducted potting experiments, bacterial growth promotion experiments, biofilm formation experiments, and chemotaxis experiments. The results showed that high concentrations of heavy metals significantly altered the rhizosphere bacterial community structure of T. repens and significantly enriched Microbacterium sp. Strain M4 was demonstrated to significantly increased the biomass and root length of T. repens under heavy metal stress. Additionally, L-proline and stigmasterol could promote bacterial growth and biofilm formation and induce chemotaxis for strain M4, suggesting that they are key rhizosphere secretions of T. repens for Microbacterium sp. recruitment. Our results suggested that T. repens adapted the heavy metal stress by reshaping rhizosphere secretions to modify the rhizosphere microbiota.
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•High concentrations of heavy metals result in significant enrichment of Microbacterium sp. in rhizosphere soil of T. repens.•Microbacterium sp. significantly increased the biomass and root length of T. repens under heavy metal stress.•L-proline and stigmasterol were key rhizosphere secretions of T. repens for recruiting Microbacterium sp.
Trifolium repens L. (T. repens) is considered a potential phytoremediation species due to its large biomass and ability to accumulate and tolerate heavy metals. Lead (Pb) is an important heavy metal ...pollutant that can affect plant growth, photosynthesis, and enzyme activity. However, response mechanism of microorganisms in three root niches of metal tolerant plants to Pb is not completely understood. Therefore, in this study, a Pb poisoning model of T. repens was established with a Pb gradient (0, 1000 mg/kg, 2000 mg/kg, and 3000 mg/kg), and was used to evaluate growth and physiological responses, as well as enrichment and transport coefficients in T. repens, and explore the characteristics of rhizosphere soil and microbial composition of three root niches. We found that Pb stress caused oxidative injury, and inhibited photosynthesis in T. repens. 16S rDNA sequencing analysis showed that the richness of microbial communities in bulk soil was higher than that in rhizosphere soil both under Pb stress and Pb nonstress conditions. Moreover, Proteobacteria was dominant phylum in bulk and rhizosphere soils, and Proteobacteria and Cyanobacteria were dominant phylum in endophytic bacteria. For the first time, we systematically investigated the response of Pb from bulk soil to plant leaves. The results showed that microbial interaction existed between bulk and rhizosphere soil. Rhizosphere bacterium Haliangium was positively correlated with urease activity and soil nutrients. Endophytic bacterium Pseudomonas was positively correlated with plant biomass and played an important role in Pb tolerance of T. repens. In addition, endophytic bacteria formed complex correlation networks with growth and physiological indexes of both root and shoot, moreover the network in root was more complicated. Taken together, Pb stress dose-dependently inhibited the growth of plants. This study provided a theoretical basis for the further development of microbial cooperation with plant remediation of heavy metal contaminated soil.
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•The richness of microbial communities in bulk soil was higher than that in rhizosphere soil both under Pb stress and Pb non stress conditions.•Proteobacteria and Cyanobacteria were dominant phylum in endophytic bacteria at all Pb levels.•Rhizosphere bacteria Haliangium was positively correlated with urease and soil nutrients under Pb pollution.•Endophytic bacterium Pseudomonas was positively correlated with plant biomass.•Correlation network formed between endophytic bacteria and growth physiological indexes in root was complex than that in shoot.
Little is known about the response of the soil microbiome (including bacteria in the rhizosphere of legumes such as clover) to mercury (Hg) despite the toxicity of Hg to soil microorganisms. Here, ...Hg-contaminated soils collected from Guizhou province, southwest China, were divided into three groups according to their Hg contents and were planted with clover. High-throughput sequencing of bacterial 16S rRNA and nitrogenase (nifH) genes and quantitative polymerase chain reaction (qPCR) were used to study the response of bacteria and diazotrophs to soil Hg stress and the effects of Hg on the abundance of functional genes in rhizosphere soils. High concentrations of soil Hg decreased bacterial community abundance and diversity and increased the abundance and diversity of nitrogen-fixing bacteria. LEfSe analysis indicates that Rhizobium was a biomarker at sites with high soil Hg contents and the co-occurrence network results indicate a positive relationship between the abundance of the dominant module (from the co-occurrence network analysis) of Rhizobiaceae and soil Hg concentration. Structural equation modeling (SEM) indicates that the Hg content in the clover shoots (ShootHg) was negatively correlated with the abundance of the mercury reductase (merA) gene (r = −0.26, P < 0.05) and the organomercury lyase (merB) gene (r = −0.23, P < 0.05) in rhizosphere soils. Moreover, correlation analysis and SEM indicate that soil total nitrogen (TN), nitrate‑nitrogen (NO3−N), soil organic matter (SOM), and available molybdenum (Mo) contents were also important factors affecting the structure of the microbial community and the abundance of functional genes. The results provide a basis for further study of the mechanism(s) by which microorganisms may impart tolerance of clover to Hg in contaminated soils.
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•High soil Hg content increased the abundance and diversity of diazotrophs.•Rhizobium acted as a biomarker at sites with high soil Hg contents.•Rhizosphere merA abundance was negatively correlated with clover shoot Hg content.
Carbon nanomaterials (CNMs) are novel engineered nanomaterials and have been used widely. Their toxic effects on terrestrial plants in soil matrix require careful investigation. In this study, white ...clover (Trifolium repens L.) was grown in a potted soil with graphene oxide (GO) at levels of 0.2%, 0.4% and 0.6% and the effects of GO on the growth and nutrient uptake of white clover were evaluated after 50 and 100 days of exposure. GO exposure showed adverse effects on seedling growth, photosynthetic parameters and nutrient uptake in shoots, and the effect was more significant with increasing concentration and exposure time. Compared with the control, GO at the highest level of 0.6% decreased plant height, leaf and stem dry weights, total chlorophyll content and net photosynthetic rate by 43.7%, 45.7%, 43.4%, 32% and 85.7%, respectively, after 100 d of exposure, and N, K, Cu, Zn, Fe, Mo, B, Si contents decreased by 19.5%, 20.1%, 12.6%, 25.0%, 12.9%, 26.0%, 18.9%, 23.0%, respectively. Furthermore, the electrolyte leakage, lipid peroxidation, reactive oxygen species, antioxidant enzyme activities were all increased by GO, especially at high dose and long exposure. These results indicate that GO can suppress plant growth by oxidative stress, photosynthesis inhibition, and nutrient imbalance.
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•Data on the toxicity of graphene oxide (GO) to white clover in soil matrix were provided.•GO adversely affected plant growth and photosynthesis.•GO induced oxidative stress in white clover.•A decrease in nutrient levels was observed in white clover after GO exposure.
Cold plasma seed treatment can promote plant growth and enhance the resistance of agricultural crops to adverse stress. However, the effects of plasma seed treatment on the growth and phytoextraction ...response of plants to cadmium (Cd) remain poorly documented. Here, we have investigated the feasibility of using plasma seed treatment to enhance the biomass and Cd accumulation of three Cd-tolerant species, namely Bidens pilosa L, Solanum nigrum L. and Trifolium repens L, under different plasma treatment conditions. Possible enhancement mechanisms are also proposed according to the levels of organic acids in the roots and the Cd fractions in rhizosphere soil following different plasma treatment conditions. The optimum plasma power was 100 W (B. pilosa) or 500 W (S. nigrum and T. repens). The optimum plasma exposure time for all three species was 60 s. Plasma seed treatment under the optimum treatment conditions enhanced plant dry biomass by ~17.3–45.0% and Cd accumulation by 8.8–54.4% across all three species compared to the controls. Furthermore, the phytoremediation efficiencies, bioaccumulation factors and transfer factors of the three species also increased significantly after seed plasma treatment. The promotion of plasma treatment on the biomass and Cd accumulation of three species might be due to increased exudation of organic acids from the roots into the rhizosphere soil, thus increasing the concentrations of acid-soluble Cd to form Cd-organic acid complexes that facilitated the uptake and translocation of Cd by the plants. Results of this study revealed that cold plasma seed treatment is an environmentally friendly, economical and efficient means to develop the application of phytoremediation for Cd-contaminated soils.
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•Cold plasma seed treatment enhanced plant biomass and Cd accumulation.•Optimum plasma power level was 100 W (B. pilosa) or 500 W (S. nigrum and T. repens).•Optimum plasma exposure time was 60 s for seeds of the three plant species.•Plasma treatment altered organic acid levels and Cd fractions in rhizosphere soils.
The invasion of alien plant and the pollution caused by soil microplastics have emerged as significant ecological threats. Recent studies have demonstrated aggravating effect of non-biodegradable ...microplastics on plant invasion. However, the impact of biodegradable microplastics (BMPs) on plant invasion remains unclear. Therefore, it is imperative to explore the impact of BMPs on plant invasion. In this study, a 30-day potting experiment with Trifolium repens L. (an invasive plant) and Oxalis corniculata L. (a native plant) was conducted to evaluate the influence of BMPs on T. repens's invasion. The findings revealed that BMPs results in a reduction in available N and P contents, thereby facilitating the colonization of arbuscular mycorrhizal fungi on T. repens 's roots. Consequently, T. repens adjusted its N and P foraging strategy by increasing P absorption ratio, and enhancing the accumulation of N and P in leaves. This ultimately led to the decrease of relative neighbor effect index of T. repens, indicating an aggravated invasion by T. repens. This study significantly enhances and expands the understanding of mechanisms by which microplastics aggravate plant invasion.
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●Biodegradable microplastics (BMPs) aggravate invasion of Trifolium repens L.●BMPs aggravate the colonization rate of AMF in invasive plant.●Invasive plants’ absorption and utilization of N and P have been altered by BMPs.
A comprehensive study of the biological effects of chronic radiation exposure (8 μGy/h) in populations of white clover (Trifolium repens L.) from the Chernobyl exclusion zone was carried out. White ...clover is one of the most important pasture legumes, having many agricultural applications. Studies at two reference and three radioactively contaminated plots showed no stable morphological effects in white clover at this level of radiation exposure. Increased activities of catalase and peroxidases were found in some impacted plots. Auxin concentration was enhanced in the radioactively contaminated plots. Genes involved in the maintenance of water homeostasis and photosynthetic processes (TIP1 and CAB1) were upregulated at radioactively contaminated plots.
•Dose rate up to 8 μGy/h is insufficient to induce stable morphological changes.•Antioxidant responses could be stimulated by chronic irradiation.•The auxin concentrations increase at the most radioactive plots.•Aquaporins and LHCII components can respond to radiation.
In grass–legume swards, biologically fixed nitrogen (N) from the legume can support the N requirements of the grass, but legume N fixation is suppressed by additional fertilizer N application. This ...study sought to identify a fertilizer N application rate that maximizes herbage and N yields, N fixation and apparent N transfer from white clover to companion grasses under intensive grazing at a site with high soil‐N status. During a 3‐year period (2011–2013), swards of perennial ryegrass and of perennial ryegrass–white clover, receiving up to 240 kg N ha−1 year−1, were compared using isotope dilution and N‐difference methods. The presence of white clover increased herbage and N yields by 12–44% and 26–72%, respectively. Applications of N fertilizer reduced sward white clover content, but the effect was less at below 120 kg N ha−1. The proportion of N derived from the atmospheric N fixation was 25–70%. Nitrogen fixation ranged from 25 to 142 kg N ha−1 measured using the isotope dilution method in 2012 and from 52 to 291 kg N ha−1 using the N‐difference method across all years. Fertilizer N application reduced the percentage and yield of fixed N. Transfer of N from white clover to grass was not confirmed, but there was an increased N content in grass and soil‐N levels. Under intensive grazing, the maximum applied N rate that optimized herbage and N yields with minimal effect on white clover content and fixation rates was 60–120 kg N ha−1.
This study was designed to determine the effect of exogenous spermidine (Spd) (30 μM) on white clover seed germination under water stress induced by polyethylene glycol 6000. Use of seed priming with ...Spd improved seed germination percentage, germination vigor, germination index, root viability and length, and shortened mean germination time under different water stress conditions. Seedling fresh weight and dry weight also increased significantly in Spd-treated seeds compared with control (seeds primed with distilled water). Improved starch metabolism was considered a possible reason for this seed invigoration, since seeds primed with Spd had significantly increased α-amylase/β-amylase activities, reducing sugar, fructose and glucose content and transcript level of β-amylase gene but not transcript level of α-amylase gene. In addition, the physiological effects of exogenous Spd on improving seeds' tolerance to water deficit during germination were reflected by lower lipid peroxidation levels, better cell membrane stability and significant higher seed vigour index in seedlings. Enhanced antioxidant enzyme activities (superoxide dismutase, peroxidase, catalase and ascorbate peroxidase), ascorbate-glutathione cycle (ASC-GSH cycle) and transcript level of genes encoding antioxidant enzymes induced by exogenous Spd may be one of the critical reasons behind acquired drought tolerance through scavenging of reactive oxygen species (ROS) in water-stressed white clover seeds. The results indicate that Spd plays an important function as a stress-protective compound or physiological activator.
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