Conferred Cd Tolerance Research Articles

Identification and characterization of cadmium stress-related LncRNAs from Betula platyphylla

Cadmium (Cd) is one of the most serious global environmental pollutants, which inhibits plant growth and interferes with their physiological processes. However, there have been few studies on the involvement of long noncoding RNAs (lncRNAs) in Cd tolerance. In the present study, we identified the lncRNAs from Betula platyphylla (birch) that respond to Cd stress. Thirty lncRNAs that were differentially expressed under Cd treatment were identified, including 16 upregulated and 14 downregulated lncRNAs. Nine differentially regulated lncRNAs were selected for further characterization. These lncRNAs were transiently overexpressed in birch plants to determine their roles in Cd tolerance. Among them, two lncRNAs conferred Cd tolerance and two induced sensitivity to Cd stress. We further determined the Cd tolerance of four target genes of the lncRNAs involved in Cd tolerance, including l-lactate dehydrogenase A (LDHA),heat shock protein (HSP18.1), yellow stripe-like protein (YSL9), and H/ACA ribonucleoprotein complex subunit 2-like protein (HRCS2L). Among them, HSP18.1 and LDHA showed improved tolerance to Cd stress, whereas the other two genes did not appear to be involved in Cd tolerance. These results suggested that lncRNAs can up- or downregulate their target genes to improve Cd tolerance. These results increase our understanding of lncRNA-mediated Cd tolerance.

Transcription Factor GmWRKY142 Confers Cadmium Resistance by Up-Regulating the Cadmium Tolerance 1-Like Genes.

Cadmium (Cd) is a widespread pollutant that is toxic to living organisms. Previous studies have identified certain WRKY transcription factors, which confer Cd tolerance in different plant species. In the present study, we have identified 29 Cd-responsive WRKY genes in Soybean [Glycine max (L.) Merr.], and confirmed that 26 of those GmWRKY genes were up-regulated, while 3 were down-regulated. We have also cloned the novel, positively regulated GmWRKY142 gene from soybean and investigated its regulatory mechanism in Cd tolerance. GmWRKY142 was highly expressed in the root, drastically up-regulated by Cd, localized in the nucleus, and displayed transcriptional activity. The overexpression of GmWRKY142 in Arabidopsis thaliana and soybean hairy roots significantly enhanced Cd tolerance and lead to extensive transcriptional reprogramming of stress-responsive genes. ATCDT1, GmCDT1-1, and GmCDT1-2 encoding cadmium tolerance 1 were induced in overexpression lines. Further analysis showed that GmWRKY142 activated the transcription of ATCDT1, GmCDT1-1, and GmCDT1-2 by directly binding to the W-box element in their promoters. In addition, the functions of GmCDT1-1 and GmCDT1-2, responsible for decreasing Cd uptake, were validated by heterologous expression in A. thaliana. Our combined results have determined GmWRKYs to be newly discovered participants in response to Cd stress, and have confirmed that GmWRKY142 directly targets ATCDT1, GmCDT1-1, and GmCDT1-2 to decrease Cd uptake and positively regulate Cd tolerance. The GmWRKY142-GmCDT1-1/2 cascade module provides a potential strategy to lower Cd accumulation in soybean.

Melatonin confers cadmium tolerance by modulating critical heavy metal chelators and transporters in radish plants.

Cadmium (Cd) is an environmental pollutant that causes health hazard to living organisms. Melatonin (MT) has emerged as a ubiquitous pleiotropic molecule capable of coordinating heavy metal (HM) stresses in plants. However, it remains unclear how melatonin mediates Cd homeostasis and detoxification at transcriptional and/or post-transcriptional levels in radish. Herein, the activities of five key antioxidant enzymes were increased, while root and shoot Cd contents were dramatically decreased by melatonin. A combined small RNA and transcriptome sequencing analysis showed that 14 differentially expressed microRNAs (DEMs) and 966 differentially expressed genes (DEGs) were shared between the Cd and Cd+MT conditions. In all, 23 and ten correlated miRNA-DEG pairs were identified in Con vs. Cd and Con vs. Cd+MT comparisons, respectively. Several DEGs encoding yellow stripe 1-like (YSL), heavy metal ATPases (HMA), and ATP-binding cassette (ABC) transporters were involved in Cd transportation and sequestration in radish. Root exposure to Cd2+ induced several specific signaling molecules, which consequently trigger some HM chelators, transporters, and antioxidants to achieve reactive oxygen species (ROS) scavenging and detoxification and eliminate Cd toxicity in radish plants. Notably, transgenic analysis revealed that overexpression of the RsMT1 (Metallothionein 1) gene could enhance Cd tolerance of tobacco plants, indicating that the exogenous melatonin confers Cd tolerance, which might be attributable to melatonin-mediated upregulation of RsMT1 gene in radish plants. These results could contribute to dissecting the molecular basis governing melatonin-mediated Cd stress response in plants and pave the way for high-efficient genetically engineering low-Cd-content cultivars in radish breeding programs.

Effects of sodium selenite on the antioxidant capacity and the fruit yield and quality of strawberry under cadmium stress

This study investigated the effects of sodium selenite (Na2SeO3) on the antioxidant capacity and the fruit yield and quality of cadmium-stressed strawberry. The results showed that cadmium (Cd) stress notably decreased the activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT) and glutathione synthetic enzyme gamma-glutamylcysteine synthetase (γ-ECS), plant height and plant biomass, and fruit yield and quality, improved the activities of ascorbate oxidase (AO), glutathione reductase (GR) and ascorbate synthetic enzyme L-galactono-1,4-lactone dehydrogenase (GalLDH), the ratios of AsA/DHA and GSH/GSSG and the contents of malondialdehyde (MDA), hydrogen peroxide (H2O2) and Cd, compared with control. Compared with Cd stress alone, 5 and 10 μM Na2SeO3 notably improved the activities of above enzymes except AO, the ratios of AsA/DHA and GSH/GSSG, plant height and biomass, Se contents in leaves and fruits, average fruit weight and the contents of Vc, soluble sugar and total soluble solid in fruits. 15 μM Na2SeO3 notably improved the activities of above enzymes except AO, the ratio of GSH/GSSG, plant height, Se contents in leaves and fruits, average fruit weight and Vc content in fruits. 20 μM Na2SeO3 only notably improved Se contents in leaves and fruits, average fruit weight and Vc content in fruits. Meanwhile, 5, 10 and 15 μM Na2SeO3 significantly reduced Cd stress-induced the contents of MDA, H2O2 and Cd, 20 μM Na2SeO3 only significantly reduced Cd stress-induced the contents of Cd in leaves and fruits, compared with Cd stress alone.Our findings indicated that only low concentrations of Se were effective in conferring Cd tolerance in strawberry plants by up-regulating their antioxidant capacity and reducing Cd accumulation, which further improved the fruit yield and quality of strawberry.

Genome-wide identification, phylogenetic and expression analysis of the heat shock transcription factor family in bread wheat (Triticum aestivum L.)

BackgroundEnvironmental toxicity from non-essential heavy metals such as cadmium (Cd), which is released from human activities and other environmental causes, is rapidly increasing. Wheat can accumulate high levels of Cd in edible tissues, which poses a major hazard to human health. It has been reported that heat shock transcription factor A 4a (HsfA4a) of wheat and rice conferred Cd tolerance by upregulating metallothionein gene expression. However, genome-wide identification, classification, and comparative analysis of the Hsf family in wheat is lacking. Further, because of the promising role of Hsf genes in Cd tolerance, there is need for an understanding of the expression of this family and their functions on wheat under Cd stress. Therefore, here we identify the wheat TaHsf family and to begin to understand the molecular mechanisms mediated by the Hsf family under Cd stress.ResultsWe first identified 78 putative Hsf homologs using the latest available wheat genome information, of which 38 belonged to class A, 16 to class B and 24 to class C subfamily. Then, we determined chromosome localizations, gene structures, conserved protein motifs, and phylogenetic relationships of these TaHsfs. Using RNA sequencing data over the course of development, we surveyed expression profiles of these TaHsfs during development and under different abiotic stresses to characterise the regulatory network of this family. Finally, we selected 13 TaHsf genes for expression level verification under Cd stress using qRT-PCR.ConclusionsTo our knowledge, this is the first report of the genome organization, evolutionary features and expression profiles of the wheat Hsf gene family. This work therefore lays the foundation for targeted functional analysis of wheat Hsf genes, and contributes to a better understanding of the roles and regulatory mechanism of wheat Hsfs under Cd stress.

Comprehensive Analysis of the Cadmium Tolerance of Abscisic Acid-, Stress- and Ripening-Induced Proteins (ASRs) in Maize.

In plants, abscisic acid-, stress-, and ripening-induced (ASR) proteins have been shown to impart tolerance to multiple abiotic stresses such as drought and salinity. However, their roles in metal stress tolerance are poorly understood. To screen plant Cd-tolerance genes, the yeast-based gene hunting method which aimed to screen Cd-tolerance colonies from maize leaf cDNA library hosted in yeast was carried out. Here, maize ZmASR1 was identified to be putative Cd-tolerant through this survival screening strategy. In silico analysis of the functional domain organization, phylogenetic classification and tissue-specific expression patterns revealed that maize ASR1 to ASR5 are typical ASRs with considerable expression in leaves. Further, four of them were cloned for testifying Cd tolerance using yeast complementation assay. The results indicated that they all confer Cd tolerance in Cd-sensitive yeast. Then they were transiently expressed in tobacco leaves for subcellular localization analysis and for Cd-challenged lesion assay, continuously. The results demonstrated that all 4 maize ASRs tested are localized to the cell nucleus and cytoplasm in tobacco leaves. Moreover, they were confirmed to be Cd-tolerance genes in planta through lesion analysis in Cd-infiltrated leaves transiently expressing them. Taken together, our results demonstrate that maize ASRs play important roles in Cd tolerance, and they could be used as promising candidate genes for further functional studies toward improving the Cd tolerance in plants.

The WRKY transcription factor, WRKY13, activates PDR8 expression to positively regulate cadmium tolerance in Arabidopsis.

Cadmium (Cd) extrusion is an important mechanism conferring Cd tolerance by decreasing its accumulation in plants. Previous studies have identified an Arabidopsis ABC transporter, PDR8, as a Cd extrusion pump conferring Cd tolerance. However, the regulation of PDR8 in response to Cd stress is still largely unknown. In this study, we identified an Arabidopsis cadmium-tolerant dominant mutant, designated xcd3-D, from the XVE-tagging T-DNA insertion lines by a gain-of-function genetic screen. The corresponding gene was cloned and shown to encode a nuclear WRKY transcription factor WRKY13. Expression of WRKY13 was induced by Cd stress. Overexpression of WRKY13 resulted in decreased Cd accumulation and enhanced Cd tolerance, whereas loss-of-function of WRKY13 led to increased Cd accumulation and sensitivity. Further analysis showed that WRKY13 activates the transcription of PDR8 by directly binding to its promoter. Genetic analysis indicated that WRKY13 acts upstream of PDR8 to positively regulate Cd tolerance. Our results provide evidence that WRKY13 directly targets PDR8 to positively regulate Cd tolerance in Arabidopsis.

Comparative analysis of Cd-responsive maize and rice transcriptomes highlights Cd co-modulated orthologs

BackgroundMetal tolerance is often an integrative result of metal uptake and distribution, which are fine-tuned by a network of signaling cascades and metal transporters. Thus, with the goal of advancing the molecular understanding of such metal homeostatic mechanisms, comparative RNAseq-based transcriptome analysis was conducted to dissect differentially expressed genes (DEGs) in maize roots exposed to cadmium (Cd) stress.ResultsTo unveil conserved Cd-responsive genes in cereal plants, the obtained 5166 maize DEGs were compared with 2567 Cd-regulated orthologs in rice roots, and this comparison generated 880 universal Cd-responsive orthologs groups composed of 1074 maize DEGs and 981 rice counterparts. More importantly, most of the orthologous DEGs showed coordinated expression pattern between Cd-treated maize and rice, and these include one large orthologs group of pleiotropic drug resistance (PDR)-type ABC transporters, two clusters of amino acid transporters, and 3 blocks of multidrug and toxic compound extrusion (MATE) efflux family transporters, and 3 clusters of heavy metal-associated domain (HMAD) isoprenylated plant proteins (HIPPs), as well as all 4 groups of zinc/iron regulated transporter protein (ZIPs). Additionally, several blocks of tandem maize paralogs, such as germin-like proteins (GLPs), phenylalanine ammonia-lyases (PALs) and several enzymes involved in JA biosynthesis, displayed consistent co-expression pattern under Cd stress.Out of the 1074 maize DEGs, approximately 30 maize Cd-responsive genes such as ZmHIPP27, stress-responsive NAC transcription factor (ZmSNAC1) and 9-cis-epoxycarotenoid dioxygenase (NCED, vp14) were also common stress-responsive genes reported to be uniformly regulated by multiple abiotic stresses. Moreover, the aforementioned three promising Cd-upregulated genes with rice counterparts were identified to be novel Cd-responsive genes in maize.Meanwhile, one maize glutamate decarboxylase (ZmGAD1) with Cd co-modulated rice ortholog was selected for further analysis of Cd tolerance via heterologous expression, and the results suggest that ZmGAD1 can confer Cd tolerance in yeast and tobacco leaves.ConclusionsThese novel findings revealed the conserved function of Cd-responsive orthologs and paralogs, which would be valuable for elucidating the genetic basis of the plant response to Cd stress and unraveling Cd tolerance genes.

Cloning, characterization and expression analysis of metallothioneins from Ipomoea aquatica and their cultivar-dependent roles in Cd accumulation and detoxification

To explore the possible roles of metallothioneins (MTs) played in cadmium (Cd) accumulation of water spinach, three IaMT genes, IaMT1, IaMT2 and IaMT3 in a high-shoot-Cd (T308) and a low-shoot-Cd accumulation cultivar (QLQ) were cloned, characterized, and quantitated. Gene expression analysis suggested that the expression of the IaMTs was differentially regulated by Cd stress in different cultivars, and T308 showed higher MTs expression overall. Furthermore, only shoot IaMT3 expression was cultivar dependent among the three IaMTs. Antioxidant analysis showed that the high production of IaMTs in T308 should be associated with its high oxidation resistance. The role of IaMTs in protecting against Cd toxicity was demonstrated in vitro via recombinant E. coli strains. The results showed that IaMT1 correlated with neither Cd tolerance nor Cd accumulation of E. coli, while IaMT2 conferred Cd tolerance in E. coli, IaMT2 and IaMT3 increased Cd accumulation in E. coli. These findings help to clarify the roles of IaMTs in Cd accumulation, and increase our understanding of the cultivar-dependent Cd accumulation in water spinach.

Iris lactea var. chinensis (Fisch.) cysteine-rich gene llCDT1 enhances cadmium tolerance in yeast cells and Arabidopsis thaliana

IlCDT1, a cysteine-rich protein, was isolated from Iris lactea var. chinensis (Fisch.) (I. lactea var. chinensis). Its transcription was up-regulated by the exogenous application of Cd. The truncated IlCDT1 (25−54) containing 14 Cys residues confers Cd tolerance to yeast as the intact IlCDT1, indicating that Cys residues are required for Cd tolerance presumably by chelating Cd. When the gene was constitutively expressed in A. thaliana, root length of transgenic lines was longer than that of wild-type under 100 μM or 200 μM Cd stress. However, Cd absorption in wild-type was more than in two trangenic lines under 100 μM Cd exposure. IlCDT1 may directly bind Cd, through chelating Cd and avoiding the Cd uptake into the cells. Together, IlCDT1 may be a promising gene for the Cd tolerance improvement. SummaryCysteine-rich gene llCDT1 enhances cadmium tolerance in yeast cells and Arabidopsis thaliana.

Wheat expansin gene TaEXPA2 is involved in conferring plant tolerance to Cd toxicity

Cadmium (Cd) is a severe and toxic heavy metal pollutant that affects plant growth and development. In this study, we found that the expression of an expansin gene, TaEXPA2, was upregulated in wheat leaves under CdCl2 toxicity. We characterized the involvement of TaEXPA2 in conferring Cd tolerance. Tobacco plants overexpressing TaEXPA2 showed higher germination rate, root elongation, and biomass accumulation compared to the wild-type (WT) plants upon CdCl2 treatment. The improved photosynthetic parameters and lesser cellular damage in transgenic plants exposed to Cd compared to that in the WT plants suggest that TaEXPA2 overexpression improves Cd tolerance in plants. Furthermore, we noticed that Cd was efficiently effluxed out of the cytoplasm in the transgenic plants owing to the enhanced activities of H+-ATPase, V-ATPase, and PPase, which helped in conferring Cd tolerance. Moreover, Cd concentration and ROS accumulation were lower in the transgenic plants than in WT plants as a consequence of enhanced antioxidant enzyme activities in the former. In addition, atexpa2, an Arabidopsis mutant, exhibited lower biomass and shorter primary root compared to its WT under Cd toxicity; however, the phenotype was recovered upon expression of TaEXPA2 in these mutants. Our results demonstrate that TaEXPA2 confers tolerance to Cd toxicity. The changed absorption/transportation of Cd and the antioxidative capacity may be involved in the improved tolerance of the transgenic plants with overexpression of TaEXPA2 to CdCl2 toxicity.

Different efficiencies of the same mechanisms result in distinct Cd tolerance within Rhizobium

Soil contamination with metals is a widespread problem posing risks to humans and ecosystems. Metal contaminated soils often hold poor microbial density and biodiversity. Among soil bacteria, rhizobia have a great agronomic and environmental significance and are major contributors to a sustainable maintenance of soil fertility. This group of microorganisms are severely affected by metals, such as cadmium (Cd), but information about metal resistance mechanisms in rhizobia is still limited. A concerted approach of the different mechanisms conferring Cd tolerance to rhizobia was conducted using two Rhizobium strains with contrasting tolerances to Cd. Results show that both strains resort to the same mechanisms (extracellular immobilization, periplasmic allocation, cytoplasmic sequestration and biotransformation of toxic products) to overcome stress, but differences in the efficiencies of some mechanisms were noticed. The ability of Rhizobium to increase glutathione in the presence of Cd emerges as a central factor in the tolerance to Cd and is as a feature to be looked for when screening or transforming microorganisms to integrate plant-microbe consortia. These could promote plant growth at contaminated sites, being more efficient for the cleanup of metals from contaminated sites and the restoration of soil quality.

Transcriptome Profiling of Abiotic Stress-Responsive Genes During Cadmium Chloride-Mediated Stress in Two Indica Rice Varieties

Rice growth and development is highly affected by the excessive concentration of the heavy metal cadmium (Cd). To elucidate the molecular basis of Cd tolerance, we carried out comprehensive transcriptome profiling of diverse groups of genes like those encoding antioxidative enzymes (APX, CAT, SOD, and GR), osmolyte-regulatory enzymes (P5CS, PDH, and BADH1), polyamine-regulatory enzymes (SAMDC, SPDS, SPMS, and DAO), transcription factors (TRAB-1 and WRKY-71), Osem and RbcS from leaf and root tissues under cadmium chloride (1.5 mM) stress at different time points (6, 12, and 24 h) of exposure in two indica rice varieties, that is, IR-64 (sensitive) and Nonabokra (tolerant). Analyses of massively parallel signature sequencing (MPSS) and publicly available microarray databases indicated the physiological role of the concerned genes and possible cross-talk between Cd and other abiotic stresses. Moreover, the pattern of expression, for a particular gene in leaves, varied in most cases from that in roots, suggesting a tissue-specific response. The higher accumulation of TRAB-1 transcript and protein in the tolerant cultivar Nonabokra in response to Cd stress suggested a role for TRAB-1 as a nodal component in Cd signaling pathway. Overall, the transcriptome analyses undertaken in the present study indicated that the genes, belonging to different functional classes and conferring Cd tolerance, were differentially regulated and mostly overrepresented under stress situations, thus providing novel insight into the functional basis of Cd tolerance in rice varieties and the importance of cross-talk of different signaling pathways. The present work will also pave the way in the future to select gene(s) for overexpression, so that rice can better tolerate Cd stress.

HsfA1a upregulates melatonin biosynthesis to confer cadmium tolerance in tomato plants.

Melatonin regulates broad aspects of plant responses to various biotic and abiotic stresses, but the upstream regulation of melatonin biosynthesis by these stresses remains largely unknown. Herein, we demonstrate that transcription factor heat-shock factor A1a (HsfA1a) conferred cadmium (Cd) tolerance to tomato plants, in part through its positive role in inducing melatonin biosynthesis under Cd stress. Analysis of leaf phenotype, chlorophyll content, and photosynthetic efficiency revealed that silencing of the HsfA1a gene decreased Cd tolerance, whereas its overexpression enhanced plant tolerance to Cd. HsfA1a-silenced plants exhibited reduced melatonin levels, and HsfA1a overexpression stimulated melatonin accumulation and the expression of the melatonin biosynthetic gene caffeic acid O-methyltransferase 1 (COMT1) under Cd stress. Both an in vitro electrophoretic mobility shift assay and in vivo chromatin immunoprecipitation coupled with qPCR analysis revealed that HsfA1a binds to the COMT1 gene promoter. Meanwhile, Cd stress induced the expression of heat-shock proteins (HSPs), which was compromised in HsfA1a-silenced plants and more robustly induced in HsfA1a-overexpressing plants under Cd stress. COMT1 silencing reduced HsfA1a-induced Cd tolerance and melatonin accumulation in HsfA1a-overexpressing plants. Additionally, the HsfA1a-induced expression of HSPs was partially compromised in COMT1-silenced wild-type or HsfA1a-overexpressing plants under Cd stress. These results demonstrate that HsfA1a confers Cd tolerance by activating transcription of the COMT1 gene and inducing accumulation of melatonin that partially upregulates expression of HSPs.

Transcription Factors PvERF15 and PvMTF-1 Form a Cadmium Stress Transcriptional Pathway.

In plants, cadmium (Cd)-responsive transcription factors are key downstream effectors of Cd stress transcriptional pathways, which are capable of converging Cd stress signals through triggering the expression of Cd detoxification genes. However, the upstream transcriptional regulatory pathways that modulate their responses to Cd are less clear. Previously, we identified the bean (Phaseolus vulgaris) METAL RESPONSE ELEMENT-BINDING TRANSCRIPTION FACTOR1 (PvMTF-1) that responds to Cd and confers Cd tolerance in planta. Here, we demonstrate an upstream transcriptional regulation of the PvMTF-1 response to Cd Using a yeast one-hybrid system, we cloned the bean ETHYLENE RESPONSE FACTOR15 (PvERF15) that binds to the PvMTF-1 promoter. PvERF15 was strongly induced by Cd stress, and its overexpression resulted in the up-regulation of PvMTF-1 DNA-protein interaction assays further revealed that PvERF15 binds directly to a 19-bp AC-rich element in the PvMTF-1 promoter. The AC-rich element serves as a positive element bound by PvERF15 to activate gene expression. More importantly, knockdown of PvERF15 by RNA interference resulted in reduced Cd-induced expression of PvMTF-1PvERF15 seems to be involved in Cd tolerance, since knockdown of PvERF15 by RNA interference in bean leaf discs decreased Cd tolerance in a transient assay. Since PvERF15 is a component of the Cd stress transcriptional pathway in beans and PvMTF-1 is one of its downstream targets, our findings provide a PvERF15/PvMTF-1 transcriptional pathway and thereby contribute to the understanding of Cd stress transcriptional regulatory pathways in plants.

Melatonin mitigates cadmium phytotoxicity through modulation of phytochelatins biosynthesis, vacuolar sequestration, and antioxidant potential in Solanum lycopersicum L.

Melatonin is a ubiquitous signal molecule, playing crucial roles in plant growth and stress tolerance. Recently, toxic metal cadmium (Cd) has been reported to regulate melatonin content in rice; however, the function of melatonin under Cd stress, particularly in higher plants, still remains elusive. Here, we show that optimal dose of melatonin could effectively ameliorate Cd-induced phytotoxicity in tomato. The contents of Cd and melatonin were gradually increased over time under Cd stress. However, such increase in endogenous melatonin was incapable to reverse detrimental effects of Cd. Meanwhile, supplementation with melatonin conferred Cd tolerance as evident by plant biomass and photosynthesis. In addition to notable increase in antioxidant enzymes activity, melatonin-induced Cd stress mitigation was closely associated with enhanced H+-ATPase activity and the contents of glutathione and phytochelatins. Although exogenous melatonin had no effect on root Cd content, it significantly reduced leaf Cd content, indicating its role in Cd transport. Analysis of Cd in different subcellular compartments revealed that melatonin increased cell wall and vacuolar fractions of Cd. Our results suggest that melatonin-induced enhancements in antioxidant potential, phytochelatins biosynthesis and subsequent Cd sequestration might play a critical role in plant tolerance to Cd. Such a mechanism may have potential implication in safe food production.

LcBiP, a endoplasmic reticulum chaperone binding protein gene from Lycium chinense, confers cadmium tolerance in transgenic tobacco

Cadmium (Cd) accumulation is very toxic to plants. The presence of Cd may lead to excessive production of reactive oxygen species (ROS), and then cause inhibition of plant growth. The endoplasmic reticulum chaperone binding protein (BiP) is an important functional protein, which has been shown to function as a sensor of alterations in the ER environment. BiP overexpression in plants was shown to increase drought tolerance through inhibition of ROS accumulation. Due to the above relationships, it is likely that there may be a link between Cd stress tolerance, ROS accumulation and the BiP transcript expression in plants. In this study, a BiP gene, LcBiP, from L. chinense was isolated and characterized. Overexpression of LcBiP in tobacco conferred Cd tolerance. Under Cd stress conditions, the transgenic tobacco lines exhibited better chlorophyll retention, less accumulation of ROS, longer root length, more glutathione (GSH) content, and less antioxidant enzyme activity than the wild type. These data demonstrated that LcBiP act as a positive regulator in Cd stress tolerance. It is hypothesized that the improved Cd tolerance of the transgenic tobacco plants may be due to the enhanced ROS scavenging capacity. The enhancement of GSH content might contribute to this ROS scavenging capacity in the transgenic plants. However, the underlying mechanism for BiP-mediated increase in Cd stress tolerance need to be further clarified.

Bean metal-responsive element-binding transcription factor confers cadmium resistance in tobacco.

Cadmium (Cd) is highly toxic to plants. Modulation of Cd-responsive transcription is an important way for Cd detoxification in plants. Metal-responsive element (MRE) is originally described in animal metallothionein genes. Although functional MREs also exist in Cd-regulated plant genes, specific transcription factors that bind MRE to regulate Cd tolerance have not been identified. Previously, we showed that Cd-inducible bean (Phaseolus vulgaris) stress-related gene2 (PvSR2) produces a short (S) PvSR2 transcript (S-PvSR2) driven by an intronic promoter. Here, we demonstrate that S-PvSR2 encodes a bean MRE-binding transcription factor1 (PvMTF-1) that confers Cd tolerance in tobacco (Nicotiana tabacum). PvMTF-1 expression was up-regulated by Cd at the levels of RNA and protein. Importantly, expression of PvMTF-1 in tobacco enhanced Cd tolerance, indicating its role in regulating Cd resistance in planta. This was achieved through direct regulation of a feedback-insensitive Anthranilate Synthase α-2 chain gene (ASA2), which catalyzes the first step for tryptophan biosynthesis. In vitro and in vivo DNA-protein interaction studies further revealed that PvMTF-1 directly binds to the MRE in the ASA2 promoter, and this binding depends on the zinc finger-like motif of PvMTF-1. Through modulating ASA2 up-regulation by Cd, PvMTF-1 increased free tryptophan level and subsequently reduced Cd accumulation, thereby enhancing Cd tolerance of transgenic tobacco plants. Consistent with this observation, tobacco transiently overexpressing ASA2 also exhibited increased tolerance to Cd. We conclude that PvMTF-1 is a zinc finger-like transcription factor that links MRE to Cd resistance in transgenic tobacco through activation of tryptophan biosynthesis.

Functional analysis of the C-terminal region of the vacuolar cadmium-transporting rice OsHMA3

Rice OsHMA3 is a vacuolar cadmium (Cd) transporter belonging to the P1B-ATPase family and has a long (273aa) C-terminal region. We analyzed the function of the region related to Cd using the transgenic Arabidopsis Col-0 ecotype, which is sensitive to Cd. The OsHMA3 variant containing a truncated (58aa) C-terminal region did not confer Cd tolerance, whereas an OsHMA3 variant containing a longer truncated (105aa) C-terminal region conferred Cd tolerance to transgenic Arabidopsis. We conclude that the C-terminal region, particularly the region containing the first 105aa, has an important role in OsHMA3 activity.

Impact of cadmium on young plants of Populus euphratica and P. × canescens, two poplar species that differ in stress tolerance

Populus euphratica is a salt tolerant species, and Populus × canescens is a salt sensitive species. Because stress tolerance is mediated by an array of overlapping defense mechanisms, we hypothesized that P. euphratica would confer co-tolerance to heavy metal stress. To test this hypothesis, both poplar species were exposed to 50 μM Cd for 24 h in hydroponic solutions. For the analyses of stress symptoms in roots and leaves, we used nutrient element concentrations and transcriptional responses of 28 stress- and defense-related genes. The roots showed strong Cd accumulation, but less in P. euphratica than in P. × canescens. In contrast, the leaves of P. euphratica accumulated 10-times more Cd than those of P. × canescens. Cd-stressed leaves of P. euphratica displayed water loss and wilting. Both species revealed contrasting transcriptional responses of defense genes to Cd stress. In P. euphratica, glutathione reductase, dehydroascorbate reductase and glutathione S-transferases of the theta and tau classes showed increases in transcript levels in response to Cd, pointing to oxidative stress and a strong activation of detoxification mechanisms. In contrast, in P. × canescens transcript levels of genes involved in signaling, activation of, and protection from drought stress were significantly increased including glutathione S-transferases of the phi class (early response to dehydration). Our data show that P. euphratica salt tolerance does not confer Cd tolerance and suggest that failure to activate early protection contributed to higher Cd sensitivity in P. euphratica than in P. × canescens.