Cadmium Tolerance Research Articles

Combined Application of 2,4-Epibrassinolide and K2SO4 Improved the Cadmium Tolerance in Tobaccos by Protecting Chloroplast and Reducing Cd Accumulation

Combined Application of 2,4-Epibrassinolide and K2SO4 Improved the Cadmium Tolerance in Tobaccos by Protecting Chloroplast and Reducing Cd Accumulation

CDR1, a DUF946 domain containing protein, positively regulates cadmium tolerance in Arabidopsis thaliana by maintaining the stability of OPT3 protein

Industrial and agricultural production processes lead to the accumulation of cadmium (Cd) in soil, resulting in crops absorb Cd from contaminated soil and then transfer it to human body through the food chain, posing a serious threat to human health. Thus, it is necessary to explore novel genes and mechanisms involved in regulating Cd tolerance and detoxification in plants. Here, we found that CDR1, a DUF946 domain containing protein, localizes to the plasma membrane and positively regulates Cd stress tolerance. The cdr1 mutants exhibited Cd sensitivity, accumulated excessive Cd in the seeds and roots, but decreased in leaves. However, CDR1-OE transgenic plants not only showed Cd tolerance but also significantly reduced Cd in seeds and roots. Additionally, both in vitro and in vivo assays demonstrated an interaction between CDR1 and OPT3. Cell free protein degradation and OPT3 protein level determination assays indicated that CDR1 could maintain the stability of OPT3 protein. Moreover, genetic phenotype analysis and Cd content determination showed that CDR1 regulates Cd stress tolerance and affect the distribution of Cd in plants by maintaining the stability of OPT3 protein. Our discoveries provide a key candidate gene for directional breeding to reduce Cd accumulation in edible seeds of crops.

Overexpression of BnaA10.WRKY75 Decreases Cadmium and Salt Tolerance via Increasing ROS Accumulation in Arabidopsis and Brassica napus L.

Soil is indispensable for agricultural production but has been seriously polluted by cadmium and salt in recent years. Many crops are suffering from this, including rapeseed, the third largest global oilseed crop. However, genes simultaneously related to both cadmium and salt stress have not been extensively reported yet. In this study, BnaA10.WRKY75 was screened from previous RNA-seq data related to cadmium and salt stress and further analyses including sequence comparison, GUS staining, transformation and qRT-PCR were conducted to confirm its function. GUS staining and qRT-PCR results indicated BnaA10.WRKY75 was induced by CdCl2 and NaCl treatment. Sequence analysis suggested BnaA10.WRKY75 belongs to Group IIc of the WRKY gene family and transient expression assay showed it was a nuclear localized transcription factor. BnaA10.WRKY75-overexpressing Arabidopsis and rapeseed plants accumulated more H2O2 and O2- and were more sensitive to CdCl2 and NaCl treatment compared with untransformed plants, which may be caused by the downregulation of BnaC03.CAT2. Our study reported that BnaA10.WRKY75 increases sensitivity to cadmium and salt stress by disrupting the balance of reactive oxygen species both in Arabidopsis and rapeseed. The results support the further understanding of the mechanisms underlying cadmium and salt tolerance and provide BnaA10.WRKY75 as a valuable gene for rapeseed abiotic stress breeding.

Inter- and intra-specific metal tolerance variation in ectomycorrhizal fungal Suillus species.

Soil metal contamination can affect growth, metabolism, and reproduction of organisms, and can lead to death. However, some fungi have evolved metal tolerance and are able to live in contaminated soils. Species in the ectomycorrhizal genus Suillus from Europe and Asia display variation in metal tolerance, yet it is unknown whether this is a widespread trait in the genus and whether it occurs in North America. Here we investigate cadmium (Cd) and zinc (Zn) tolerance in S. brevipes and S. tomentosus isolates collected from sites in the Rocky Mountains of Colorado displaying different metal content. In line with previous findings for other Suillus species, we hypothesized (1) S. brevipes and S. tomentosus to display intra-specific metal tolerance variation, (2) Zn and Cd tolerance to be correlated to soil metal content, and (3) tolerant isolates to show lower metal tissue content compared to sensitive isolates (due to increased metal exclusion). We found ample intra- and inter-specific Zn and Cd tolerance variation in both S. brevipes and S. tomentosus, but no correlation between soil metal content and tolerance. There was a negative correlation between tolerance level and Zn uptake, indicating an exclusion-based Zn tolerance strategy. Sensitive and tolerant isolates showed no difference in Cd accumulation, indicating that Cd tolerance in these species is likely not dependent on exclusion. Our study sets the groundwork for further investigation into the genetic basis of Suillus metal tolerance and whether and how it impacts pine mycorrhizal partners.

In vitro exploration of Acinetobacter strain (SG-5) for antioxidative potential and phytohormone biosynthesis in maize (Zea mays L.) cultivars differing in cadmium tolerance.

Cadmium (Cd) poses serious threats to plant growth and development, whereas the use of plant growth-promoting rhizobacteria (PGPR) has emerged a promising approach to diminish Cd retention in crops. A pot experiment was conducted to evaluate the effect of Cd tolerant strain Acinetobacter sp. SG-5 on growth, phytohormonal response, and Cd uptake of two maize cultivars (3062 and 31P41) under various Cd stress levels (0, 5, 12, 18, 26, and 30μM CdCl2). The results revealed that CdCl2 treatment significantly suppressed the seed germination and growth together with higher Cd retention in maize cultivars in a dose-dependent and cultivar-specific manner with pronounced negative effect in 31P41. However, SG-5 strain exerted positive impact by up-regulating seed germination traits, plant biomass, photosynthetic pigments, enzymatic and non-enzymatic antioxidants, endogenous hormone level indole-3-acetic acid (IAA), abscisic acid (ABA), and sustained optimal nutrient's levels in both cultivars but predominantly in Cd-sensitive one (31P41). Further, Cd-resistant PGPR decreased the formation of reactive oxygen species in terms of malondialdehyde (MDA) and hydrogen peroxide (H2O2) verified through 3, 3'-diaminobenzidine (DAB) and nitroblue tetrazolium (NBT) analysis in conjunction with reduced Cd uptake and translocation in maize root and shoots in comparison to controls, advocating its sufficiency for bacterial-assisted Cd bioremediation. In conclusion, both SG-5 inoculated cultivars exhibited maximum Cd tolerance but substantial Cd tolerance was acquired by Cd susceptible cultivar-31P41 than Cd-tolerant one (3062). Current work recommended SG-5 strain as a promising candidate for plant growth promotion and bacterial-assisted phytomanagement of metal-polluted agricultural soils.

Poly-glutamic acid reinforces wheat cadmium tolerance by modulating ascorbic acid and glutathione metabolism

Poly-glutamic acid reinforces wheat cadmium tolerance by modulating ascorbic acid and glutathione metabolism

A comparative study on cadmium tolerance and applicability of two Solanum lycopersicum L. cultivars.

Solanum lycopersicum L. can be classified into low Cd-accumulating and high Cd-accumulating types based on their accumulation characteristics of cadmium (Cd). There are many common S. lycopersicum varieties available in the market, but their specific Cd tolerance and enrichment abilities are not well understood. This article uses two S. lycopersicum cultivars, Yellow Cherry and Yellow Pearl, as experimental materials. The experimental method of soil pot planting was adopted, and Cd concentrations in the soil were added at 0, 0.6, 1.5, 2.5, 5, and 10 mg/kg. The changes in Cd content, biomass, photosynthetic pigment content, and photosynthetic parameters of the two S. lycopersicum cultivars were analyzed to screen for low-accumulation S. lycopersicum cultivars. The results showed that S. lycopersicum are Cd-sensitive plants. The Cd accumulation, photosynthetic parameters, and other basic indicators of Yellow Cherry basically showed significant differences when the soil Cd concentration was 0.6 mg/kg, and the biomass showed significant differences when the soil Cd concentration was 1.5 mg/kg. Except for the Cd accumulation in the roots and leaves of Yellow Pearl, which showed significant differences at a soil Cd concentration of 0.6 mg/kg, the other indicators basically showed significant differences when the soil Cd concentration was 1.5 mg/kg. When the soil Cd concentration was 0.6 mg/kg, the Cd accumulation in the fruit of Yellow Pearl was 0.04 mg/kg, making it a low-accumulation S. lycopersicum variety suitable for promoting cultivation in Cd-contaminated soil at 0.6 mg/kg. In conclusion, the Cd accumulation in the fruit of Yellow Pearl is significantly lower than that of Yellow Cherry and even below the Cd limit value for fresh vegetables specified in GB2762-2017. Therefore, Yellow Pearl can be grown as edible crops in soils with Cd concentrations ≤0.6 mg/kg. Furthermore, Yellow Cherry demonstrate strong Cd tolerance and can be used for the remediation of Cd-contaminated soils.

Metabolomics analysis of bahia grass (Paspalum notatum) inoculated with arbuscular mycorrhizal fungi exposed to soil Cd stress

Heavy metals can adversely affect the growth and metabolic processes of plant. Arbuscular mycorrhizal (AM) fungi, which colonize the roots of most plants, may change the uptake and resistance to heavy metals by altering plant metabolism. But few published papers focused on how AM fungi enhanced plant tolerance to heavy metals at the metabolomics level. Therefore, our study incorporated LC-MS technology to explore the molecular mechanism by which AM fungi improved cadmium (Cd) tolerance of Bahia grass (Paspalum notatum) at the metabolomic scale. The results showed that AM fungi increased growth of Bahia grass, and enhanced its uptake and tolerance of Cd. Infection with AM fungi significantly raised the levels of both primary metabolites (amino acid, carbohydrate, organic acid, etc.) and secondary metabolites (such as terpenoids, phenolic compounds, and alkaloids), which in turn improved the photosynthesis efficiency, osmoregulation, and antioxidation defense of the plant, thereby enhancing the Cd tolerance of Bahia grass. Furthermore, under high concentration of Cd, both TCA cycle and lipid metabolism were generally suppressed, and AM fungi inoculation alleviated the effects and normalized the TCA cycle and lipid metabolism. In all, the metabolic alterations identified here provided insights into the mechanisms by which AM fungi enhanced plant tolerance to heavy metals.

Root cell wall polysaccharides and endodermal barriers restrict long-distance Cd translocation in the roots of Kentucky bluegrass

Soil Cd pollution is a significant environmental issue faced by contemporary society. Kentucky bluegrass is considered a potential phytoremediation species, as some varieties have excellent cadmium (Cd) tolerance. However, the mechanisms of Cd accumulation and transportation in Kentucky bluegrass are still not fully understood. The Cd-tolerant Kentucky bluegrass cultivar ‘Midnight’ (M) exhibits lower Cd translocation efficiency and a higher leaf Cd concentration compared to the Cd-sensitive cultivar ‘Rugby II’ (R). We hypothesized that Cd translocation from roots to shoots in cultivar M is hindered by the endodermal barriers and cell wall polysaccharides; hence, we conducted Cd distribution, cytological observation, cell wall component, and transcriptomic analyses under Cd stress conditions using the M and R cultivars. Cd stress resulted in the thickening of the endodermis and increased synthesis of cell wall polysaccharides in both the M and R cultivars. Endodermis development restricted the radical transport of Cd from the root cortex to the stele, while the accumulation of cell wall polysaccharides promoted the binding of Cd to the cell wall. These changes further inhibited the long-distance translocation of Cd from the roots to the aerial parts. Furthermore, the M cultivar exhibited limited long-distance Cd translocation efficiency compared to the R cultivar, which was attributed to the enhanced development of endodermal barriers and increased Cd binding by cell wall polysaccharides. This study provides valuable insights for screening high Cd transport efficiency in Kentucky bluegrass based on anatomical structure and genetic modification.

Cross-protection and cross-feeding between Enterobacter and Comamonas promoting their coexistence and cadmium tolerance in Oryza sativa L.

Metabolic cross-feeding is a pervasive interaction between bacteria to acquire novel phenotypes. However, our current understanding of the survival mechanism for cross-feeding in cocultured bacterial biofilms under heavy-metal conditions remains limited. Herein, we found that Comamonas sp. A23 produces L-phenylalanine to activate the L-phenylalanine degradation pathway in Enterobacter sp. A11, enhancing biofilm formation and cadmium [Cd(II)] immobilization in A11. The genes responsible for L-phenylalanine-degradation (paaK) and cell attachment and aggregation (csgAD) are essential for biofilm formation and Cd(II) immobilization in A11 induced by L-phenylalanine. The augmentation of A11 biofilms, in turn, protects A23 under Cd(II) and H2O2 stresses. The plant-based experiments demonstrate that the induction of two rice Cd(II) transporters, OsCOPT4 and OsBCP1, by A11 and A23 enhances rice resistance against Cd(II) and H2O2 stresses. Overall, our findings unveil the mutual dependence between bacteria and rice on L-phenylalanine cross-feeding for survival under abiotic stress.

Overexpression of histone demethylase gene SlJMJ18 and SlJMJ23 from tomato confers cadmium tolerance by regulating metal transport and hormone content in Arabidopsis

A lower concentration of cadmium (Cd), a hazardous and non-essential element for plant growth, will have deleterious effects on plants and endanger human health. Histone demethylase (JHDM) is important for plants' ability to withstand abiotic stress, according to an increasing number of studies. The degree of expression of the SlJMJ18 and SlJMJ23 genes in different tomato tissues was confirmed by this study. These two genes were responsive to the heavy metals Cd, Hg, Pb, and Cu stress, according to fluorescence quantification and GUS staining. Interestingly, the overexpression transgenic Arabidopsis plants of two genes have different responses to Cd stress. While SlJMJ18-OE lines consistently display Cd resistance but an early-flowering phenotype, SlJMJ23-OE plants have sensitivity during the post-germination stage and then greater tolerance to Cd stress. It was discovered that these two genes may affect cadmium tolerance of plants by regulating the expression of hormone synthesis related genes and hormone contents (BRs and ABA). Moreover, SlJMJ23 may resist cadmium stress by increasing the total phenol content in plants. The functional significance of JMJs is better understood in this study, which also offers a theoretical foundation for the use of molecular technology to develop plants resistant to Cd and an experimental basis for the efficient use of land resources.

Microwave-assisted method to morpholinyl analogue and its effect on cadmium tolerance of soybean

A series of morpholiny analogues were efficiently obtained through nucleophilic aromatic substitution (SNAr) under microwave irradiation with simple operation. Furthermore, the molecular docking studies suggested that the compound 4m could bind receptor PYR1 much more strongly than abscisic acid. Significantly, the application of 4m could reduce the permeability of cell membranes to enhance the tolerance. Furthermore, the growth of root and above-ground parts and the fresh weight was improved when treatment with 4m under cadmium stress at 200–500 mg/L. Thus, compound 4m was successfully designed and synthesized as the most potent candidate.

Physiological and transcriptome characterization provide new sights into cadmium tolerance and accumulation mechanisms in Tagetes patula L.

Tagetes patula L. (T. patula) is a Cd hyperaccumulator plant with high ornamental value and strong resistance. Understanding the molecular mechanisms of Cd tolerance in hyper-accumulating plants is important for improving phytoremediation efficiency. This investigation studied to how Cd affected T. patula's physiological characteristics and molecular alterations. With the increase of Cd concentration, chlorophyll content, and root biomass decreased; the activities of antioxidant enzymes such as GST, SOD and POD increased. Initially, Cd accumulated mainly in the root and then transferred upward to the leaves. The molecular mechanisms of T. patula after 150 mg/kg and 250 mg/kg treatments were analyzed by transcriptome. In total, 5429 and 6546 differentially expressed unigenes were identified. Phenylpropanoid biosynthesis and phenylalanine metabolism were identified as key pathways in reaction to Cd stress. In addition, many differential genes involved in the response to Cd stress were identified, including heavy metal transporter protein genes, genes and transcription factors related to Cd detoxification. These results indicate that the response of T. patula to Cd was multifaceted, laying the foundation for studies to improve its tolerance to toxic metals.

Metabolic regulation mechanism of melatonin for reducing cadmium accumulation and improving quality in rice

Melatonin acts as a potential regulator of cadmium (Cd) tolerance in rice. However, its practical value in rice production remains unclear. To validate the hypothesis that melatonin affects Cd accumulation and rice quality, a series of experiments were conducted. The results showed that exogenous melatonin application was associated with reduced Cd accumulation (23–43%) in brown rice. Fourier transform infrared spectroscopy (FTIR) analysis showed that exogenous melatonin affected the rice protein secondary structure and starch short-range structure. Metabolomics based on LC–MS/MS revealed that exogenous melatonin altered the brown rice metabolic profile, decreased fatty acid metabolite content, but increased amino acid metabolite, citric acid, melatonin biosynthetic metabolite, and plant hormone contents. These findings indicate that exogenous melatonin can effectively reduced Cd accumulation and improve rice quality through metabolic network regulation, serving as an effective treatment for rice cultivated in Cd-contaminated soil.

Integrated proteomics, transcriptomics, and metabolomics offer novel insights into Cd resistance and accumulation in Poa pratensis

Kentucky bluegrass (Poa pratensis L., KB) demonstrates superior performance in both cadmium (Cd) accumulation and tolerance; however, the regulatory mechanisms and detoxification pathways in this species remain unclear. Therefore, phenotype, root ultrastructure, cell wall components, proteomics, transcriptomics, and metabolomics were analyzed under the hydroponic system to investigate the Cd tolerance and accumulation mechanisms in the Cd-tolerant KB variety ‘Midnight (M)’ and the Cd-sensitive variety ‘Rugby II (R)’ under Cd stress. The M variety exhibited higher levels of hydroxyl and carboxyl groups as revealed by Fourier transform infrared spectroscopy spectral analysis. Additionally, a reduced abundance of polysaccharide degradation proteins was observed in the M variety. The higher abundance of glutathione S-transferase and content of L-cysteine-glutathione disulfide and oxidized glutathione in the M variety may contribute to better performance of the M variety under Cd stress. Additionally, the R variety had an enhanced content of carboxylic acids and derivatives, increasing the Cd translocation capacity. Collectively, the down-regulation of cell wall polysaccharide degradation genes coupled with the up-regulation of glutathione metabolism genes enhances the tolerance to Cd stress in KB. Additionally, lignification of the endodermis and the increase in carboxylic acids and derivatives play crucial roles in the redistribution of Cd in KB.

The impact of elevated sulfur and nitrogen levels on cadmium tolerance in Euglena species

Heavy metal (HM) pollution threatens human and ecosystem health. Current methods for remediating water contaminated with HMs are expensive and have limited effect. Therefore, bioremediation is being investigated as an environmentally and economically viable alternative. Freshwater protists Euglena gracilis and Euglena mutabilis were investigated for their tolerance to cadmium (Cd). A greater increase in cell numbers under Cd stress was noted for E. mutabilis but only E. gracilis showed an increase in Cd tolerance following pre-treatment with elevated concentrations of S or N. To gain insight regarding the nature of the increased tolerance RNA-sequencing was carried out on E. gracilis. This revealed transcript level changes among pretreated cells, and additional differences among cells exposed to CdCl2. Gene ontology (GO) enrichment analysis reflected changes in S and N metabolism, transmembrane transport, stress response, and physiological processes related to metal binding. Identifying these changes enhances our understanding of how these organisms adapt to HM polluted environments and allows us to target development of future pre-treatments to enhance the use of E. gracilis in bioremediation relating to heavy metals.

Unveiling silicon-mediated cadmium tolerance mechanisms in mungbean (Vigna radiata (L.) Wilczek): Integrative insights from gene expression, antioxidant responses, and metabolomics

Cadmium (Cd), one of the most phytotoxic heavy metals, is a major contributor to yield losses in several crops. Silicon (Si) is recognized for its vital role in mitigating Cd toxicity, however, the specific mechanisms governing this mitigation process are still not fully understood. In the present study, the effect of Si supplementation on mungbean (Vigna radiata (L.) Wilczek) plants grown under Cd stress was investigated to unveil the intricate pathways defining Si derived stress tolerance. Non-invasive leaf imaging technique revealed improved growth, biomass, and photosynthetic efficiency in Si supplemented mungbean plants under Cd stress. Further, physiological and biochemical analysis revealed Si mediated increase in activity of glutathione reductase (GR), ascorbate peroxidase (APX), and catalase (CAT) enzymes involved in reactive oxygen species (ROS) metabolism leading to mitigation of cellular damage and oxidative stress. Untargeted metabolomic analysis using liquid chromatography coupled with mass spectrometry (LC-MS/MS) provided insights into Si mediated changes in metabolites and their respective pathways under Cd stress. Alteration in five different metabolic pathways with major changes in flavanols and flavonoids biosynthesis pathway which is essential for controlling plants antioxidant defense system and oxidative stress management were observed. The information reported here about the effects of Si on photosynthetic efficiency, antioxidant responses, and metabolic changes will be helpful in understanding the Si-mediated resistance to Cd stress in plants.

Phytoremediation potential of Brassica oleracea varieties through cadmium tolerance gene expression analysis

BackgroundBrassica oleracea var. acephala, commonly referred to as kale, is a well-documented plant species, a food crop but well recognized for its capacity to endure and manage the accumulation of heavy metals. In this research, the phytoremediation potential of kale was evaluated based on cadmium intake, utilizing three distinct kale varieties originating from Bosnia and Herzegovina. All kales were grown in controlled conditions, with different concentrations of cadmium (Cd), a known strong pollutant found in small concentrations in soil under normal environmental conditions. After the root length analysis and cadmium atomic spectrometry, we utilized quantitative PCR (qPCR) and cycle threshold (Ct) values to calculate the expression levels of five genes associated with Cd heavy metal response: Mitogen-activated protein kinase 2 (MAPK2), Farnesylated protein 26 and 27 (HIPP26, HIPP27), Natural resistance-associated macrophage protein 6 (RAMP6), and Heavy metal accumulator 2 (HMA2). ResultsThe atomic reader's analysis of rising cadmium concentrations revealed a proportional decline in the length of kale roots. The gene expression levels corresponded to cadmium stress differently among varieties, but mostly showing notable up-regulations under Cd stress, indicating the strong Cd presence within the plant. ConclusionsThis study demonstrated differences in gene expression behavior among three B. oleracea varieties from Bosnia and Herzegovina, indicating and filtering the Cd-resistant kale, and kale varieties suitable for phytoremediation. For the first time, such a study was conducted on kale varieties from Bosnia and Herzegovina, analyzing the impact of cadmium on the growth and resilience of these species.

Salicylic Acid's impact on Sedum alfredii growth and cadmium tolerance: Comparative physiological, transcriptomic, and metabolomic study

With the acceleration of industrialization, Cd pollution has emerged as a major threat to soil ecosystem health and food safety. Hyperaccumulating plants like Sedum alfredii Hance are considered to be used as part of an effective strategy for the ecological remediation of Cd polluted soils. This study delved deeply into the physiological, transcriptomic, and metabolomic responses of S. alfredii under cadmium (Cd) stress when treated with exogenous salicylic acid (SA). We found that SA notably enhanced the growth of S. alfredii and thereby increased absorption and accumulation of Cd, effectively alleviating the oxidative stress caused by Cd through upregulation of the antioxidant system. Transcriptomic and metabolomic data further unveiled the influence of SA on photosynthesis, antioxidant defensive mechanisms, and metal absorption enrichment pathways. Notably, the interactions between SA and other plant hormones, especially IAA and JA, played a central role in these processes. These findings offer us a comprehensive perspective on understanding how to enhance the growth and heavy metal absorption capabilities of hyperaccumulator plants by regulating plant hormones, providing invaluable strategies for future environmental remediation efforts.

Transcriptomic analysis provides insights into the molecular mechanism of melatonin-mediated cadmium tolerance in Medicago sativa L.

Cadmium (Cd), a toxic element, often makes a serious threat to plant growth and development. Previous studies found that melatonin (Mel) reduced Cd accumulation and reestablished the redox balance to alleviate Cd stress in Medicago sativa L., however, the complex molecular mechanisms are still elusive. Here, comparative transcriptome analysis and biochemical experiments were conducted to explore the molecular mechanisms of Mel in enhancing Cd tolerance. Results showed that 7237 differentially expressed genes (DEGs) were regulated by Mel pretreatment to Cd stress compared to the control condition in roots of Medicago sativa L. Besides, in comparison with Cd stress alone, Mel upregulated 1081 DEGs, and downregulated 1085 DEGs. These DEGs were mainly involved in the transcription and translation of genes and folding, sorting and degradation of proteins, carbohydrate metabolism, and hormone signal network. Application of Mel regulated the expression of several genes encoding ribosomal protein and E3 ubiquitin-protein ligase involved in folding, sorting and degradation of proteins. Moreover, transcriptomic analyse suggested that Mel might regulate the expression of genes encoding pectin lyase, UDP-glucose dehydrogenase, sucrose-phosphate synthase, hexokinase-1, and protein phosphorylation in the sugar metabolism. Therefore, these could promote sucrose accumulation and subsequently alleviate the Cd damage. In conclusion, above findings provided the mining of important genes and molecular basis of Mel in mitigating Cd tolerance and genetic cultivation of Medicago sativa L.