Cadmium Detoxification Research Articles

Cadmium (Cd) detoxification and activation of plant defense enzymes activation in Wheat (Triticum aestivum) through the use of endophytic Bacillus thuringiensis and Salix alba root powder

Cadmium (Cd) is a toxic heavy metal and a threat to the ecosystem therefore the current investigation was designed to use endophytic bacteria from the Salix alba roots and to investigate its plant growth promoting and Cd detoxification ability with and without Salix alba root powder. In a complete randomized design (CRD), the cadmium sulfate was applied at the rate of 20mg/kg and 40mg/kg soil. The Bacillus thuringiensis (Accession # MW979616) was identified from Salix alba roots. The combination of Bacillus thuringiensis inoculated seeds +0.5gm root powder showed significant increase in wheat shoot dry weight, root fresh weight, catalase, and ascorbate peroxidases by 457%, 223%, 105% and 74%, respectively. The application of Bacillus thuringiensis with Salix alba root powder boosted the plant growth and defense at higher concentrations of Cd. In another treatment with Bacillus thuringiensis inoculated seeds + CdSO4 40mg/kg + 0.5gm root powder significantly increased the shoot fresh weight, root fresh weight, root dry weight, proline, sugar, superoxide dismutase, and peroxidase by 456%, 650%, 115%, 91%, 80%, 350%, and 250%, respectively with 80% reduction in plant Cd accumulation and increased bacterial population. Bacillus thuringiensis and Salix alba root powder can be useful for plant growth, Cd toxicity mitigation, accelerating bacterial activity in Cd-contaminated soil and uplifting the plant defense under heavy metal stress.

Wood biochar induced metal tolerance in Maize (Zea mays L.) plants under heavy metal stress

Due to metal toxicity, widespread industrialization has negatively impacted crop yield and soil quality. The current study was aimed to prepare and characterize biochar made from wood shavings of Pinus roxburghii and to determine the plant growth promoting and heavy metal detoxification of cadmium (Cd) and chromium (Cr) contaminated soil. FTIR SEM coupled with EDX characterization of biochar was performed; Cd and Cr were used at a rate of 20 mg/kg. Biochar was used at the rate of 50 mg/kg for various treatments. The completely randomized design (CRD) was used for the experiment and three replicates of each treatment were made. Various agronomic and enzymatic parameters were determined. The results indicated that all growth and enzymatic parameters were enhanced by the prepared biochar treatments. The most prominent results were observed in treatment T5 (in which shoot length, root length, peroxidase dismutase (POD), superoxide dismutase (SOD) catalyzes (CAT), and chlorophyll a and b increased by 28%, 23%, 40%, 41%, 42%, and 27%, respectively, compared to the control). This study demonstrated that biochar is a sustainable and cost-effective approach for the remediation of heavy metals, and plays a role in plant growth promotion. Farmers may benefit from the current findings, as prepared biochar is easier to deliver and more affordable than chemical fertilizers. Future research could clarify how to use biochar optimally, applying the minimum amount necessary while maximizing its benefits and increasing yield.

Combinatorial accumulation, stress response, detoxification and synaptic transmission effects of cadmium and selenium in clams Ruditapes philippinarum

This study investigated the toxicological effects and mechanisms of cadmium (Cd) (5 and 50 μg/L) and selenium (Se) (3 and 30 μg/L) at environmentally relevant concentrations on the gills and digestive glands of clams Ruditapes philippinarum. Results indicated that Cd and Se could tissue-specifically impact osmoregulation, energy metabolism, and synaptic transmission in the gills and digestive glands of clams. After exposure to 50 μg/L Cd, the digestive glands of clams up-regulated the expression of methionine-gamma-lyase and metallothionein for detoxification. Clam digestive glands exposed to 3 μg/L Se up-regulated the expression of catalase and glutathione peroxidase to alleviate oxidative stress, and down-regulated the expression of selenide-water dikinase to reduce the conversion of inorganic Se. Additionally, the interaction mode between Cd and Se largely depended on their molar ratio, with a ratio of 11.71 (50 μg/L Cd + 3 μg/L Se) demonstrated to be particularly harmful, as manifested by significantly more lesions, oxidative stress, and detoxification demand in clams than those exposed to Cd or Se alone. Collectively, this study revealed the complex interaction patterns and mechanisms of Cd and Se on clams, providing a reference for exploring their single and combined toxicity.

The Uptake, Transfer, and Detoxification of Cadmium in Plants and Its Exogenous Effects.

Cadmium (Cd) exerts a toxic influence on numerous crucial growth and development processes in plants, notably affecting seed germination rate, transpiration rate, chlorophyll content, and biomass. While considerable advances in Cd uptake and detoxification of plants have been made, the mechanisms by which plants adapt to and tolerate Cd toxicity remain elusive. This review focuses on the relationship between Cd and plants and the prospects for phytoremediation of Cd pollution. We highlight the following issues: (1) the present state of Cd pollution and its associated hazards, encompassing the sources and distribution of Cd and the risks posed to human health; (2) the mechanisms underlying the uptake and transport of Cd, including the physiological processes associated with the uptake, translocation, and detoxification of Cd, as well as the pertinent gene families implicated in these processes; (3) the detrimental effects of Cd on plants and the mechanisms of detoxification, such as the activation of resistance genes, root chelation, vacuolar compartmentalization, the activation of antioxidant systems and the generation of non-enzymatic antioxidants; (4) the practical application of phytoremediation and the impact of incorporating exogenous substances on the Cd tolerance of plants.

Integrated physiological, transcriptomic and metabolomic analyses provide insights into phosphorus-mediated cadmium detoxification in Salix caprea roots

Phosphorus (P) plays a crucial role in facilitating plant adaptation to cadmium (Cd) stress. However, the molecular mechanisms underlying P-mediated responses to Cd stress in roots remain elusive. This study investigates the effects of P on the growth, physiology, transcriptome, and metabolome of Salix caprea under Cd stress. The results indicate that Cd significantly inhibits plant growth, while sufficient P alleviates this inhibition. Under Cd exposure, P sufficiency resulted in increased Cd accumulation in roots, along with reduced oxidative stress levels (superoxide anion and hydrogen peroxide contents were reduced by 16.8% and 30.1%, respectively). This phenomenon can be attributed to the enhanced activities of antioxidant enzymes such as superoxide dismutase (SOD) and catalase (CAT), as well as increased levels of antioxidants including ascorbic acid (AsA) and flavonoids under sufficient P conditions. A total of 4208 differentially expressed genes (DEGs) and 552 differentially accumulated metabolites (DAMs) were identified in the transcriptomic and metabolomic analyses, with 2596 DEGs and 113 DAMs identified among treatments with different P levels under Cd stress, respectively. Further combined analyses reveal the potential roles of several pathways in P-mediated Cd detoxification, including flavonoid biosynthesis, ascorbate biosynthesis, and plant hormone signal transduction pathways. Notably, sufficient P upregulates the expression of genes including HMA, ZIP, NRAMP and CAX, all predicted to localize to the cell membrane. This may elucidate the heightened Cd accumulation under sufficient P conditions. These findings provide insights into the roles of P in enhancing plant resistance to Cd stress and improving of phytoremediation.

Multilevel Regulation of Membrane Proteins in Response to Metal and Metalloid Stress: A Lesson from Yeast.

In the face of flourishing industrialization and global trade, heavy metal and metalloid contamination of the environment is a growing concern throughout the world. The widespread presence of highly toxic compounds of arsenic, antimony, and cadmium in nature poses a particular threat to human health. Prolonged exposure to these toxins has been associated with severe human diseases, including cancer, diabetes, and neurodegenerative disorders. These toxins are known to induce analogous cellular stresses, such as DNA damage, disturbance of redox homeostasis, and proteotoxicity. To overcome these threats and improve or devise treatment methods, it is crucial to understand the mechanisms of cellular detoxification in metal and metalloid stress. Membrane proteins are key cellular components involved in the uptake, vacuolar/lysosomal sequestration, and efflux of these compounds; thus, deciphering the multilevel regulation of these proteins is of the utmost importance. In this review, we summarize data on the mechanisms of arsenic, antimony, and cadmium detoxification in the context of membrane proteome. We used yeast Saccharomyces cerevisiae as a eukaryotic model to elucidate the complex mechanisms of the production, regulation, and degradation of selected membrane transporters under metal(loid)-induced stress conditions. Additionally, we present data on orthologues membrane proteins involved in metal(loid)-associated diseases in humans.

Combined application of biochar and metal-tolerant bacteria alleviates cadmium toxicity by modulating the antioxidant defense mechanism and physicochemical attributes in rice (Oryza sativa L.) grown in cadmium-contaminated soil

Agricultural soil contamination by heavy metal (cadmium) is a widespread issue and causes deleterious effects on crops. One of the most effective and ecofriendly methods for cadmium detoxification is microbe-assisted phytoremediation and soil amendment with biochar. The current study investigated the combined effect of biochar (Parthenium hysterophorus L.) and microbes (Bacillus cereus strain B2) in alleviating cadmium toxicity in rice grown in cadmium-contaminated soil (0, 10, 15, and 20 mg/kg). Results revealed that cadmium caused damage to plant health. Maximum reduction in plant biomass, gas exchange parameters, and chlorophyll content was observed on cadmium 20 mg/kg. As compared with the sole application of biochar (2 %) and microbes, the greatest plant height (33.68%), photosynthetic rate (23.52 %), stomatal conductance (48.93 %), water use efficiency (21.41 %), transpiration rate (58.87 %), chlorophyll a (74.81 %), chlorophyll b (31.28 %), total chlorophyll (60.53 %), and carotenoids (26.97 %) were recorded with combined application of biochar and microbes. Similarly, superoxidase (SOD), catalase (CAT), peroxidase (POD), and absorbate peroxidase (APX) activity was significantly boosted by combined application of microbes and biochar (2 %), which were 33.90 %, 24.18 %, 51.70 %, and 19.06 %, respectively, while significant reduction was found in the ROS (16.60 %), (20.06 %) and MDA (26.08 %) content in rice plants. However, cadmium concentrations in different parts of plant were decreased at higher concentrations of biochar (2 %) with microbes. These findings indicate that combined application of Bacillus cereus strain B2 and biochar could be a promising strategy for reducing rice crop cadmium toxicity and boosting its growth, physiology, and defense system efficiently as compared with sole application.

Juggling cadmium detoxification and zinc homeostasis: A division of labour between the two C. elegans metallothioneins

The chemical properties of toxic cadmium and essential zinc are very similar, and organisms require intricate mechanisms that drive selective handling of metals. Previously regarded as unspecific “metal sponges”, metallothioneins (MTLs) are emerging as metal selectivity filters. By utilizing C. elegans mtl-1 and mtl-2 knockout strains, metal accumulation in single worms, single copy fluorescent-tagged transgenes, isoform specific qPCR and lifespan studies it was possible to demonstrate that the handling of cadmium and zinc by the two C. elegans metallothioneins differs fundamentally: the MTL-2 protein can handle both zinc and cadmium, but when it becomes unavailable, either via a knockout or by elevated cadmium exposure, MTL-1 takes over zinc handling, leaving MTL-2 to sequester cadmium. This division of labour is reflected in the folding behaviour of the proteins: MTL-1 folded well in presence of zinc but not cadmium, the reverse was the case for MTL-2. These differences are in part mediated by a zinc-specific mononuclear His3Cys site in the C-terminal insertion of MTL-1; its removal affected the entire C-terminal domain and may shift its metal selectivity towards zinc. Overall, we uncover how metallothionein isoform-specific responses and protein properties allow C. elegans to differentiate between toxic cadmium and essential zinc.

Chromate-induced methylglyoxal detoxification system drives cadmium and chromate immobilization by Cupriavidus sp. MP-37

The detoxification of cadmium (Cd) or chromium (Cr) by microorganisms plays a vital role in bacterial survival and restoration of the polluted environment, but how microorganisms detoxify Cd and Cr simultaneously is largely unknown. Here, we isolated a bacterium, Cupriavidus sp. MP-37, which immobilized Cd(II) and reduced Cr(VI) simultaneously. Notably, strain MP-37 exhibited variable Cd(II) immobilization phenotypes, namely, cell adsorption and extracellular immobilization in the co-presence of Cd(II) and Cr(VI), while cell adsorption in the presence of Cd(II) alone. To unravel Cr(VI)-induced extracellular Cd(II) immobilization, proteomic analysis was performed, and methylglyoxal-scavenging protein (glyoxalase I, GlyI) and a regulator (YafY) showed the highest upregulation in the co-presence of Cd(II) and Cr(VI). GlyI overexpression reduced the intracellular methylglyoxal content and increased the immobilized Cd(II) content in extracellular secreta. The addition of lactate produced by GlyI protein with methylglyoxal as substrate increased the Cd(II) content in extracellular secreta. Reporter gene assay, electrophoretic mobility shift assay, and fluorescence quenching assay demonstrated that glyI expression was induced by Cr(VI) but not by Cd(II), and that YafY positively regulated glyI expression by binding Cr(VI). In the pot experiment, inoculation with the MP-37 strain reduced the Cd content of Oryza sativa L., and their secreted lactate reduced the Cr accumulation in Oryza sativa L. This study reveals that Cr(VI)-induced detoxification system drives methylglyoxal scavenging and Cd(II) extracellular detoxification in Cd(II) and Cr(VI) co-existence environment.

Wound healing and Cadmium detoxification in the earthworm Lumbricus terrestris - a potential case for coelomocytes?

Earthworms are affected by physical stress, like injury, and by exposure to xenobiotics, such as the toxic metal cadmium (Cd), which enters the environment mainly through industry and agriculture. The stress response to the single and the combination of both stressors was examined in regenerative and unharmed tissue of Lumbricus terrestris to reveal if the stress response to a natural insult like injury (amputation) interferes with Cd detoxification mechanisms. We characterized the roles of metallothionein 1 (MT1) and MT2 isoforms, heat shock protein 70 as well as immune biomarkers such as the toll-like receptors (TLR) single cysteine cluster TLR and multiple cysteine cluster TLR. The role of the activated transcription factors (ATFs) ATF2, ATF7, and the cAMP responsive element binding protein as putative regulatory intersection as well as a stress-dependent change of the essential trace elements zinc and calcium was analyzed. Phosphorylated AMP activated protein kinase, the cellular energy sensor, was measured to explore the energy demand, while the energy status was determined by detecting carbohydrate and protein levels. Taken together, we were able to show that injury rather than Cd is the driving force that separates the four treatment groups - Control, Cd exposure, Injury, Cd exposure and injury. Interestingly, we found that gene expression differed regarding the tissue section that was analyzed and we hypothesize that this is due to the migration of coelomocytes, earthworm immune cells, that take over a key role in protecting the organism from a variety of environmental challenges. Surprisingly, we discovered a role for MT1 in the response to multiple stressors and an isoform-specific function for the two newly characterized TLRs. In conclusion, we gathered novel information on the relation of innate immunity, wound healing, and Cd detoxification mechanisms in earthworms.

Intervening Role of Tyrosine in Cadmium Detoxification, Balancing of Mineral Ions Homeostasis, Antioxidants, and Secondary Metabolites in Maize

Intervening Role of Tyrosine in Cadmium Detoxification, Balancing of Mineral Ions Homeostasis, Antioxidants, and Secondary Metabolites in Maize

Multiple insights into lignin-mediated cadmium detoxification in rice (Oryza sativa)

Cadmium (Cd) is readily absorbed by rice and enters the food chain, posing a health risk to humans. A better understanding of the mechanisms of Cd-induced responses in rice will help in developing solutions to reduce Cd uptake in rice. Therefore, this research attempted to reveal the detoxification mechanisms of rice in response to Cd through physiological, transcriptomic and molecular approaches. The results showed that Cd stress restricted rice growth, led to Cd accumulation and H2O2 production, and resulted cell death. Transcriptomic sequencing revealed glutathione and phenylpropanoid were the major metabolic pathways under Cd stress. Physiological studies showed that antioxidant enzyme activities, glutathione and lignin contents were significantly increased under Cd stress. In response to Cd stress, q-PCR results showed that genes related to lignin and glutathione biosynthesis were upregulated, whereas metal transporter genes were downregulated. Further pot experiment with rice cultivars with increased and decreased lignin content confirmed the causal relationship between increased lignin and reduced Cd in rice. This study provides a comprehensive understanding of lignin-mediated detoxification mechanism in rice under Cd stress and explains the function of lignin in production of low-Cd rice to ensure human health and food safety.

The detoxification of cadmium in Japanese quail by pomegranate peel powder

ABSTRACT Environmental pollution and exposure to toxic metals such as cadmium (Cd) can cause severe and chronic diseases and have significant side effects on vital organs. The present study aimed to evaluate the effect of pomegranate peel on biochemical factors and lipid peroxidation in intoxication by Cd in Japanese quail. Two hundred seventy quails in different groups were fed diets containing Cd and pomegranate peel from 6 to 35 days old. Then, serum biochemical parameters were assessed, including liver enzymes, urea, and thiobarbituric acid. In the quails, Cd significantly increased MDA, urea, and AST (P < 0.05). Adding pomegranate peel at 1.5 and 2% levels decreased these parameters significantly (P < 0.05). In conclusion, dietary enrichment using pomegranate peel reduced the adverse effects of Cd by improving lipid peroxidation, aspartate aminotransferase (AST), and urea in Japanese quail.

Cadmium-Responsive Expression and Activation of Cytosol-Localized Sugar Beet BvGSTU9 Associated with Cadmium Detoxification

Cadmium-Responsive Expression and Activation of Cytosol-Localized Sugar Beet BvGSTU9 Associated with Cadmium Detoxification

Glutathione and phytochelatins jointly allow intracellular and extracellular detoxification of cadmium in the liverwort Marchantia polymorpha

Plants have evolved a set of mechanisms that control and respond to the uptake and accumulation of both essential and non-essential metals, including chelation and sequestration of these elements by thiol ligands, such as glutathione and phytochelatins. Indeed, such thiol peptides can chelate some metals, quickly form thiol-metal complexes, and segregate them in the vacuolar compartment. Reasonably, conceptually similar mechanisms can be assumed to be responsible for the transport of metal complexes ̶ in particular thiol-cadmium complexes ̶ across the plasma membrane, with the consequent release of this toxic metal in the extracellular environment. Such hypothesis, focusing on prevention and detoxification mechanisms, was here verified in axenically-grown gametophytes of the model liverwort Marchantia polymorpha, exposed to three different cadmium concentrations over five exposure times. From the data obtained, it can be deduced that the cell wall of M. polymorpha moderately reduced the influx of cadmium into the cells, since it was rapidly saturated by this metal. At an intracellular level, cadmium induced the activity, but not the gene expression, of the phytochelatin synthase enzyme, leading to synthesis of phytochelatins. Moreover, both glutathione and phytochelatins chelated cadmium at the cytosolic level and allowed its detoxification, possibly involving tonoplast transporters belonging to type-C ABC subfamily (Mp7g13860 and Mp4g11930). Likewise, cadmium, glutathione and phytochelatins were released extracellularly, thus highlighting a possible novel role in cadmium detoxification, in a pH-dependent manner. The overall results suggest that, in M. polymorpha, glutathione and phytochelatins can accomplish intracellular and extracellular detoxification of cadmium.

SpHsfA4c from Sedum plumbizincicola Enhances Cd Tolerance by the AsA–GSH Pathway in Transgenic Populus × canescens

The ascorbate (AsA)–glutathione (GSH) metabolism pathway is an important antioxidant system in cadmium (Cd) detoxification; the AsA–GSHpathway is generally regulated by a specific set of functional genes. However, transcription factors involved in AsA–GSH pathway have yet to be identified. Herein, we transformed a heat shock transcription factor SpHsfA4c from Sedum plumbizincicola into Populus. × canescens. Under 100 μM CdCl2 stress for 30 d, the leaf chlorosis of wild-type poplars (WT) is more serious than that in transgenic poplars. The root biomass, shoot biomass and tolerance index (TIs) of transgenic poplars were higher than those in WT. In addition, transgenic poplars have higher Cd2+ uptake and Cd content. Compared with WT, the contents of hydrogen peroxide (H2O2) and superoxide anion (O2•−) in transgenic poplars were significantly reduced in leaves under Cd treatment. The expression levels of five enzymes (ascorbate peroxidase (APX), catalases (CAT), superoxide dismutase (SOD), peroxidase (POD) and glutathione S-transferase (GST)) were higher in transgenic poplars than those in WT. Transgenic poplars contained higher concentrations of intermediate metabolites, including GSH, AsA and phytochelatins (PCs), and a higher GSH/GSSG ratio in the AsA–GSH metabolism pathway. In Fourier transform infrared (FTIR) spectra, the characteristic peaks indicated that the contents of cysteine, GSH and AsA in transgenic poplars were exceeded compared to those in WT. These results suggested that SpHsfA4c can activate the AsA–GSH metabolism pathway to reduce Cd-associated oxidative stress. Therefore, overexpressing SpHsfA4c in P. × canescens can give rise to a superior Cd tolerance. Our results provide a theoretical significance for breeding potential new germplasm resources with high biomass and high Cd tolerance for remediation of soil heavy metal pollution.

Metabolic profiles in the xylem sap of Brassica juncea exposed to cadmium.

Metabolic profiles in xylem sap are considered a fundamental mechanism for Cadmium (Cd) detoxification in plants. However, the metabolic mechanism of Brassica juncea xylem sap in response to Cd is still unclear. Here, we investigated the effects on the metabolomics of B. juncea xylem sap treated with Cd at different times by utilizing a nontargeted liquid chromatography-mass spectrometry (LC-MS)-based metabolomics method for further elucidating the response mechanism of Cd exposure. The findings indicated that 48 h and 7 days Cd exposure caused significant differences in metabolic profiles of the B. juncea xylem sap. Those differential metabolites are primarily involved in amino acids, organic acids, lipids, and carbohydrates, and most of them were downregulated, which played essential roles in response to Cd stress. Furthermore, B. juncea xylem sap resisted 48-h Cd exposure via regulation of glycerophospholipid metabolism, carbon metabolism, aminoacyl-tRNA biosynthesis, glyoxylate and dicarboxylate metabolism, linoleic acid metabolism, C5-branched dibasic acid metabolism, alpha-linolenic acid metabolism, cyanoamino acid metabolism, ABC transporters, biosynthesis of amino acids, and pyrimidine metabolism; whereas alpha-linolenic acid metabolism, glycerophospholipid metabolism, photosynthesis, and oxidative phosphorylation were regulated for resisting 7-day Cd exposure.

A novel cadmium detoxification pathway in Tri-spine horseshoe crab (Tachypleus tridentatus): A 430-million-years-ago organism

Marine and intertidal heavy metal pollution has been a major concern in recent years. Tachypleus tridentatus has existed on earth for more than 430 million years. It has suffered a sharp decline in population numbers caused by environmental pollution and anthropogenic disturbance for almost 40 years. However, the effects of heavy metal pollution on juvenile T. tridentatus have not been reported. Here we show the mechanism of cadmium (Cd) detoxification in juvenile T. tridentatus using integrated antioxidant indexes and transcriptomic and metabolomic analysis. High Cd2+ concentration caused oxidative stress in juvenile T. tridentatus. The hazards increase with increasing Cd2+ concentration in juvenile T. tridentatus. Transcriptomics and metabolomics analyses concluded that high Cd2+ concentration resulted in the imbalance of glycerophospholipid metabolism in juvenile T. tridentatus to detoxify Cd. Our results offer a rationale for protective measures and further studies of heavy metal stress in T. tridentatus.

Phytochelatins: Sulfur-Containing Metal(loid)-Chelating Ligands in Plants.

Phytochelatins (PCs) are small cysteine-rich peptides capable of binding metal(loid)s via SH-groups. Although the biosynthesis of PCs can be induced in vivo by various metal(loid)s, PCs are mainly involved in the detoxification of cadmium and arsenic (III), as well as mercury, zinc, lead, and copper ions, which have high affinities for S-containing ligands. The present review provides a comprehensive account of the recent data on PC biosynthesis, structure, and role in metal(loid) transport and sequestration in the vacuoles of plant cells. A comparative analysis of PC accumulation in hyperaccumulator plants, which accumulate metal(loid)s in their shoots, and in the excluders, which accumulate metal(loid)s in their roots, investigates the question of whether the endogenous PC concentration determines a plant's tolerance to metal(loid)s. Summarizing the available data, it can be concluded that PCs are not involved in metal(loid) hyperaccumulation machinery, though they play a key role in metal(loid) homeostasis. Unraveling the physiological role of metal(loid)-binding ligands is a fundamental problem of modern molecular biology, plant physiology, ionomics, and toxicology, and is important for the development of technologies used in phytoremediation, biofortification, and phytomining.

Retracted: Effects of Free Amino Acids, Metallothionein, and Chemical Forms of Heavy Metals on the Accumulation and Detoxification of Cadmium and Chromium in Chinese Goldthread (Coptis chinensis Franch.)

Retracted: Effects of Free Amino Acids, Metallothionein, and Chemical Forms of Heavy Metals on the Accumulation and Detoxification of Cadmium and Chromium in Chinese Goldthread (<i>Coptis chinensis</i> Franch.)