Cadmium Accumulation In Shoots Research Articles

Root cell-type specific expressions of bacterial mercury transporter MerC and plant SNARE SYP121 fusion protein differentially affect cadmium accumulation patterns of Arabidopsis

ABSTRACT There is increasing demand for solutions against cadmium pollution to secure food safety, and phytoremediation is one of the potential tools. We previously found that a bacterial mercury transporter MerC possesses cadmium uptake activity and its overexpression as a fusion protein with a plasma-membrane resident SNARE protein, SYP121 enhances the cadmium uptake ability of Arabidopsis plants. In this study, we examined whether two different root cell-type specific expression systems of MerC-SYP121 fusion protein could efficiently enhance cadmium accumulation of Arabidopsis plants, compared to the p35S-driven ubiquitous expression system. Representative transgenic lines expressing MerC-SYP121 in root surface cells (pEpi lines) or root endodermal cells (pSCR lines), established in our previous studies, were subjected to different cadmium treatments along with the p35S line. A vertical agar plate assay showed that root surface-specific line pEpi, as well as the p35S line, showed about 15% higher cadmium accumulation in shoots after one-day 10 µM cadmium treatment, compared to the wild-type Col-0. On the other hand, the endodermis-specific line pSCR accumulated 30% less cadmium in its shoots. A similar cadmium accumulation pattern in shoots was observed under the environmentally relevant much lower cadmium treatment of 0.1 µM for 4 d, using the hydroponic culture system. To further examine the potential of the MerC-SYP121 expression system for cadmium phytoremediation, the transgenic plants were hydroponically exposed to 0.1 µM for 4 weeks. The cadmium accumulation after the 4 weeks of treatment was again 16% higher in the pEpi shoots compared to that of Col-0, whereas the p35S line only showed 6% higher Cd concentration. Shoots of the pSCR line accumulated slightly less cadmium compared to Col-0. Ionomic profiles in these plants were analyzed, however, pSCR-specific patterns were not evident. Nevertheless, in our previous studies, pEpi and pSCR lines both efficiently accumulated more mercury in shoots than in the wild-type. The presented results suggest that the effects of cell-type-specific MerC-SYP121 expression differ by the target metals and its expression in root surface cells rather than that in endodermis is suitable for enhancing root cadmium uptake and subsequent shoot accumulation.

Overexpression of OsLCT2, a Low-Affinity Cation Transporter Gene, Reduces Cadmium Accumulation in Shoots and Grains of Rice

Cadmium (Cd)-contaminated rice is a serious issue affecting food safety. Understanding the molecular regulatory mechanisms of Cd accumulation in rice grains is crucial to minimizing Cd concentrations in grains. We identified a member of the low-affinity cation transporter family, OsLCT2 in rice. It was a membrane protein. OsLCT2 was expressed in all tissues of the elongation and maturation zones in roots, with the strongest expression in pericycle and stele cells adjacent to the xylem. When grown in Cd-contaminated paddy soils, rice plants overexpressing OsLCT2 significantly reduced Cd concentrations in the straw and grains. Hydroponic experiment demonstrated its overexpression decreased the rate of Cd translocation from roots to shoots, and reduced Cd concentrations in xylem sap and in shoots of rice. Moreover, its overexpression increased Zn concentrations in roots by up-regulating the expression of OsZIP9, a gene responsible for Zn uptake. Overexpression of OsLCT2 reduces Cd accumulation in rice shoots and grains by limiting the amounts of Cd loaded into the xylem and restricting Cd translocation from roots to shoots of rice. Thus, OsLCT2 is a promising genetic resource to be engineered to reduce Cd accumulation in rice grains.

Responses of soil bacterial community and Cd phytoextraction to a Sedum alfredii-oilseed rape (Brassica napus L. and Brassica juncea L.) intercropping system

Soil pollution with heavy metals has become a common problem in agricultural ecosystems and poses a threat to food safety and human health. Intercropping is now considered a promising alternative to address this issue. However, our understandings about the influences of intercropping systems on rhizosphere microbiota composition and their association with plant performance are still limited. In this study, rhizobox microcosm experiments were conducted to investigate the influence of cropping regimes (i.e. monoculture and intercropping) on the rhizosphere bacterial microbiota and their linkages with the phytoextraction of cadmium (Cd) by Zhongyouza 19 (Brassica napus L.), Xikou Huazi (Brassica juncea L.) and Sedum alfredii using 16S rRNA gene sequencing. Cadmium accumulation in shoots of B. napus and B. juncea grown under intercropping were enhanced by 370% and 27.8% respectively, as compared to monoculture. Soil compartmentation as a major determinant explained 57.6% of the rhizosphere bacterial microbiota variation, whereas plant species and cropping regime accounted for 26.4% of the variation. The overall abundance of the taxa was Proteobacteria, Acidobacteria, Bacteroidetes, Chloroflexi, Verrucomicrobia, and Actinobacteria. Intercropping significantly enriched amplicon sequence variants (ASVs) abundance belonging to Actinobacteria, Bacilli, Deltaproteobacteria while depleting that of Acidobacteria in rhizosphere. Intercropping with S. alfredii influenced more on microbial composition of B. napus rhizosphere. The change in rhizosphere bacterial communities was related to metal availability, soil properties, and plant parameters. The enriched families of Pedosphaeraceae, Ruminococcaceae, Chitinophagaceae, Gemmatimonadaceae, Nitrosomonadaceae, and Parachlamydiaceae were positively correlated with metal concentration in plants. These results indicate that S. alfredii and oilseed rape intercropping could be a promising approach for enhancing the remediation of Cd contaminated soil. Understanding the complex plant-microbe-metal interactions of intercropping system could facilitate the development of remediation strategy for phytoremediation of contaminated soils and sustainable agricultural production.

Veronica becabonga L. as a hyperaccumulatorplant for cadmium

Heavy metal pollution of water is a major environmental problem facing the modem world.The major objective of this research was to evaluate the potential of water-speedwell plant,Veronica becabonga L. to uptake and accumulate heavy metal cadmium under greenhouseconditions, Veronica becabonga L., were cultured in 3% Hoagland's nutrient medium, whichwas supplemented with 0, 25, 50,100, 200, 300 mg/I of Cd(NO3)2 4H2O over one weektreatment period. Plants were harvested at the end of this period and heavy metals from theentire shoot tissue was extracted using the closed Teflon vessel method and metal content inthe extract was estimated using a Flame atomic absorption spectrophotometry. The resultsshowed that the uptake and accumulation of Cd in V becabonga L. showed significantincrease when metal concentration was increased. The highest amount of Cd accumulationwas detected at 100mg/I Cd(NO3)24H2O in the culture solution that was 20660.3 mg/kg DWof shoots. The linear pattern of uptake suggest the involvement of both active and passivetransport mechanisms for Cd uptake, Finally, since the high concentrations cadmiumaccumulation in shoots of plants has far exceeded 0.01% DW V becabonga L. is ahyperaccumulator plant for this metal and has potential for phytoremediation of watercontaminated with cadmium,

Homology modeling and in vivo functional characterization of the zinc permeation pathway in a heavy metal P-type ATPase.

The P1B ATPase heavy metal ATPase 4 (HMA4) is responsible for zinc and cadmium translocation from roots to shoots in Arabidopsis thaliana. It couples ATP hydrolysis to cytosolic domain movements, enabling metal transport across the membrane. The detailed mechanism of metal permeation by HMA4 through the membrane remains elusive. Here, homology modeling of the HMA4 transmembrane region was conducted based on the crystal structure of a ZntA bacterial homolog. The analysis highlighted amino acids forming a metal permeation pathway, whose importance was subsequently investigated functionally through mutagenesis and complementation experiments in plants. Although the zinc pathway displayed overall conservation among the two proteins, significant differences were observed, especially in the entrance area with altered electronegativity and the presence of a ionic interaction/hydrogen bond network. The analysis also newly identified amino acids whose mutation results in total or partial loss of the protein function. In addition, comparison of zinc and cadmium accumulation in shoots of A. thaliana complemented lines revealed a number of HMA4 mutants exhibiting different abilities in zinc and cadmium translocation. These observations could be instrumental to design low cadmium-accumulating crops, hence decreasing human cadmium exposure.

Genetic Analysis of Cadmium Accumulation in Shoots of Sorghum Landraces

Contamination of agricultural fields with heavy metals, such as Cd, is a serious issue for safe food production worldwide. Previous reports indicate that sorghum [Sorghum bicolor (L.) Moench] is useful for phytoextraction of Cd from contaminated soils. To investigate the diversity for Cd accumulation in sorghum germplasm and to select useful cultivars for phytoextraction, we conducted experiments using 106 sorghum landraces and the cultivar BTx623, grown in hydroponic cultures and under field conditions. The Cd concentrations in plant shoots of different landraces showed no correlation between hydroponic and field‐grown plants (r = −0.16). We measured Cd concentrations in various plant parts at different stages of development in field‐grown plants and found that leaf sheaths often had higher Cd concentrations than other plant parts. Our results indicated that screening could be done for plants that might be useful for phytoextraction of Cd by determining the Cd concentrations in leaf sheaths of 1‐mo‐old plants grown in a Cd‐contaminated soil. We also mapped the quantitative trait loci (QTLs) responsible for Cd accumulation in sorghum shoots and found several chromosomal regions that affected Cd concentrations in different parts of the shoot. We were able to identify several promising landraces for developing sorghum cultivars that are capable of accumulating high levels of Cd and phytoextracting Cd from contaminated soils.

A loss-of-function allele of OsHMA3 associated with high cadmium accumulation in shoots and grain of Japonica rice cultivars.

Excessive cadmium (Cd) accumulation in rice poses a risk to food safety. OsHMA3 plays an important role in restricting Cd translocation from roots to shoots. A non-functional allele of OsHMA3 has been reported in some Indica rice cultivars with high Cd accumulation, but it is not known if OsHMA3 allelic variation is associated with Cd accumulation in Japonica cultivars. In this study, we identified a Japonica cultivar with consistently high Cd accumulation in shoots and grain in both field and greenhouse experiments. The cultivar possesses an OsHMA3 allele with a predicted amino acid mutation at the 380(th) position from Ser to Arg. The haplotype had no Cd transport activity when the gene was expressed in yeast, and the allele did not complement a known nonfunctional allele of OsHMA3 in F1 test. The allele is present only in temperate Japonica cultivars among diversity panels of 1483 rice cultivars. Different cultivars possessing this allele showed greatly increased root-to-shoot Cd translocation and a shift in root Cd speciation from Cd-S to Cd-O bonding determined by synchrotron X-ray absorption spectroscopy. Our study has identified a new loss-of-function allele of OsHMA3 in Japonica rice cultivars leading to high Cd accumulation in shoots and grain.

Xylem transport and gene expression play decisive roles in cadmium accumulation in shoots of two oilseed rape cultivars (Brassica napus)

Cadmium (Cd) is a toxic metal which harms human health through food chains. The mechanisms underlying Cd accumulation in oilseed rape are still poorly understood. Here, we investigated the physiological and genetic processes involved in Cd uptake and transport of two oilseed rape cultivars (Brassica napus). L351 accumulates more Cd in shoots but less in roots than L338. A scanning ion-selective electrode technique (SIET) and uptake kinetics of Cd showed that roots were not responsible for the different Cd accumulation in shoots since L351 showed a lower Cd uptake ability. However, concentration-dependent and time-dependent dynamics of Cd transport by xylem showed L351 exhibited a superordinate capacity of Cd translocation to shoots. Additionally, the Cd concentrations of shoots and xylem sap showed a great correlation in both cultivars. Furthermore, gene expression levels related to Cd uptake by roots (IRT1) and Cd transport by xylem (HMA2 and HMA4) were consistent with the tendencies of Cd absorption and transport at the physiological level respectively. In other words, L351 had stronger gene expression for Cd transport but lower for Cd uptake. Overall, results revealed that the process of Cd translocation to shoots is a determinative factor for Cd accumulation in shoots, both at physiological and genetic levels.

Reduction of Cadmium Uptake of Rice Plants Using Soil Amendments in High Cadmium Contaminated Soil: A Pot Experiment

The aims of this study were to investigate the effect of agricultural residues on reducing cadmium uptake in rice plants. The rice plants growing on no cadmium/free cadmium soils (N), Cd soils (Cds), and Cd soils each amended with 1% w/w of coir pith (CP), coir pith modified with sodium hydroxide (CPm) and corncob (CC) under high cadmium contaminated soil with an average 145 mg Cd kg -1 soil were investigated. The results showed that the cumulative transpiration of rice grown in various treatments under high cadmium contaminated soil followed the order: Cds > CPm ≥ CP ≥ CC. These transpirations directly influenced cadmium accumulation in shoots and husks of rice plants. The CC and CP seemed to work to reduce the cadmium uptake by rice plants indicated by accumulated cadmium in the husk that were 2.47 and 7.38 mg Cd kg -1 dry weight, respectively. Overall, transpiration tended to drive cadmium accumulation in plants for rice grown in high cadmium contaminated soil. The more that plants uptake cadmium, the lower cadmium that remains in the soil. Keywords : Transpiration, Cadmium Uptake, Rice plant, Agricultural Residues

Cadmium Accumulation and Its Effects on Uptake of Micronutrients in Indian Mustard [Brassica juncea (L.) Czern.] Grown in a Loamy Sand Soil Artificially Contaminated with Cadmium

A pot experiment was conducted in a greenhouse to evaluate the effects of different levels of cadmium (Cd) on Cd accumulation and their effects on uptake of micronutrients in Indian mustard [Brassica juncea (L.) Czern.]. Cadmium accumulation in shoots and interactions among other metals [manganese (Mn), iron (Fe), copper (Cu), and zinc (Zn)] were investigated. Ten levels of Cd ranging from 0 to 200 mg kg–1 soil were tested. The crop was grown for 60 days in a loamy sand soil with adequate basal fertilization of nitrogen (N), phosphorus (P), and potassium (K), and dry-matter yield (DMY) was recorded. The plants were analyzed for total Cd and micronutrients, and the soil was analyzed for diethylenetriaminepentaacetic acid (DTPA)–extractable Cd. Experimental results showed that the DTPA-extractable Cd in the soil increased consistently and significantly with increase in rates of Cd application up to 200 mg Cd kg–1 soil. Significant reduction in the DMY of Indian mustard occurred with application of 5 mg Cd kg–1 soil and greater. The content as well as uptake of Cd by Indian mustard increased significantly over the control at all rates of its application. It increased from 5.95 μg pot–1 in the control to 150.6 μg pot–1 at Cd application of 200 mg kg–1 soil. Application of Cd to soil though decreased the content of micronutrients in plants, but significant reduction occurred only for Fe at rates beyond 50 mg Cd kg–1 soil. However, the total removal of Fe, Zn, and Cu registered a significant decline over the control at and above Cd application of 10 mg kg–1 and that of Mn beyond 10 mg kg–1. In loamy sand soil, a DTPA-extractable Cd level of 3.8 mg kg–1 soil and in plant content of 28.0 μg Cd g–1 DMY was found to be the upper threshold levels of Cd for Indian mustard. Considerable residual content in the soil suggests that once the soil is contaminated by Cd it remains available in the soil for decades, and food crops grown on these soils may be a significant source of Cd toxicity to both humans and grazing animals.

The Brassica juncea BjCdR15, an ortholog of Arabidopsis TGA3, is a regulator of cadmium uptake, transport and accumulation in shoots and confers cadmium tolerance in transgenic plants

*A bZIP transcription factor from Brassica juncea (BjCdR15) was isolated by the cDNA-amplified fragment length polymorphism technique after cadmium treatment. Sequence analysis indicated high similarity between BjCdR15 and Arabidopsis TGA3. In Arabidopsis, TGA3 transcription is also induced by cadmium; hence, we investigated whether BjCdR15 is involved in cadmium tolerance and whether it can functionally replace TGA3 protein in Arabidopsis tga3-2 mutant plants. *BjCdR15 expression was detected mainly in the epidermis and vascular system of cadmium-treated plants, and increased in roots and leaves after cadmium treatment. The overexpression of BjCdR15 in Arabidopsis and tobacco enhanced cadmium tolerance: overexpressing plants showed high cadmium accumulation in shoots. Conversely, Arabidopsis tga3-2 mutant plants showed high cadmium content in roots and inhibition of its transport to the shoot. *We demonstrated that BjCdR15 can functionally replace TGA3: in 35S::BjCdR15-tga3-2 plants, the long-distance transport of cadmium from root to shoot was restored and these plants showed an increased cadmium content in shoots compared with all other assays. In addition, BjCdR15/TGA3 regulated the synthesis of phytochelatin synthase and the expression of several metal transporters. *The results indicate that BjCdR15/TGA3 transcription factors play a crucial role in the regulation of cadmium uptake by roots and in its long-distance root to shoot transport. BjCdR15/TGA3 may thus be considered as useful candidates for potential biotechnological applications in the phytoextraction of cadmium from polluted soils.

Effects of simultaneous expression of heterologous genes involved in phytochelatin biosynthesis on thiol content and cadmium accumulation in tobacco plants

Transgenic tobacco (Nicotiana tabacum cv. LA Burley 21) lines expressing three genes encoding enzymes thought to be critical for the efficient production of phytochelatins, (i) serine acetyltransferase (EC 2.3.1.30) involved in the production of O-acetylserine, the cysteine precursor, (ii) gamma-glutamylcysteine synthetase (EC 6.3.2.2) involved in the production of gamma-glutamylcysteine, the precursor of glutathione, and (iii) phytochelatin synthase (EC 2.3.2.15), were obtained and analysed for non-protein thiol content and cadmium accumulation. After a 3 week exposure to 15 microM CdCl2, plants expressing transgenes (either separately or in combination) had increased cadmium concentration in roots but not in shoots compared with the wild type. Nearly all transgenic lines analysed had more non-protein thiols than the wild type. The greatest effects (about 8-fold elevation of thiols) were found in one of the lines simultaneously expressing the three transgenes. Despite the fact that a multi-transgene strategy described in this work resulted in a strong increase in the levels of several classes of non-protein thiols in transgenic plants, other factors appeared to restrict cadmium accumulation in shoots.