Metal Tolerance Protein Research Articles

Identification and evolutionary analysis of the metal-tolerance protein family in eight Cucurbitaceae species.

Metal-tolerance proteins (MTPs) are divalent cation transporters and play fundamental roles in plant metal tolerance and ion homeostasis. Despite that, a systematic investigation of MTPs in Cucurbitacea is still lacking. In this study, 142 MTPs were identified from 11 released genomes of 8 Cucurbitaceae species. They were phylogenetically separated into three clusters (Zn-cation diffusion facilitator proteins [CDFs], Fe/Zn-CDFs, and Mn-CDFs) and further subdivided into seven groups (G1, G5, G6, G7, G8, G9, and G12). Characterization analysis revealed that most MTPs were plasma membrane-located hydrophobic proteins. Motif and exon/intron analysis showed that members in the same group contained similar conserved motifs and gene structures. Moreover, 98 pairs of segmental-like duplication events were found. The nonsynonymous/synonymous substitution ratios between each pair were less than 1, implying that Cucurbitaceae MTPs were under purification selection. Expression profiling suggested that several MTP genes, such as CsCLMTP1, CmeMTP3, LsMTP3, and Cl97103MTP3, were constitutively expressed in corresponding Cucurbitaceae species, and their expression levels were not significantly altered by NaCl, drought, or pathogen infection. The expression patterns of cucumber MTP genes under Zn2+ , Cu2+ , Mn2+ , and Cd2+ stress were studied by quantitative real-time polymerase chain reaction and the results showed that these MTPs were induced by at least one metal ion, suggesting their involvement in metal tolerance or transportation. The identification and comprehensive investigation of MTP family members will provide a basis for the analysis of ion transport functions and ion tolerance mechanisms of Cucurbitaceae MTPs.

The cell biology of zinc.

Nearly 10% of all plant proteins belong to the zinc (Zn) proteome. They require Zn either for catalysis or as a structural element. Most of the protein-bound Zn in eukaryotic cells is found in the cytosol. The fundamental differences between transition metal cations in the stability of their complexes with organic ligands, as described by the Irving-Williams series, necessitate buffering of cytosolic Zn (the 'free Zn' pool) in the picomolar range (i.e. ~6 orders of magnitude lower than the total cellular concentration). Various metabolites and peptides, including nicotianamine, glutathione, and phytochelatins, serve as Zn buffers. They are hypothesized to supply Zn to enzymes, transporters, or the recently identified sensor proteins. Zn2+ acquisition is mediated by ZRT/IRT-like proteins. Metal tolerance proteins transport Zn2+ into vacuoles and the endoplasmic reticulum, the major Zn storage sites. Heavy metal ATPase-dependent efflux of Zn2+ is another mechanism to control cytosolic Zn. Spatially controlled Zn2+ influx or release from intracellular stores would result in dynamic modulation of cellular Zn pools, which may directly influence protein-protein interactions or the activities of enzymes involved in signaling cascades. Possible regulatory roles of such changes, as recently elucidated in mammalian cells, are discussed.

Identification and characterization of Metal Tolerance Protein (MTP) family in Glycyrrhiza uralensis

Identification and characterization of Metal Tolerance Protein (MTP) family in Glycyrrhiza uralensis

Metal tolerance protein MTP6 is involved in Mn and Co distribution in poplar

With the booming demand of the electric vehicle industry, the concentration of manganese (Mn) and cobalt (Co) flowing into land ecosystems has also increased significantly. While these transition metals can promote the growth and development of plants, they may become toxic under high concentrations. It is thus important to understand how Mn and Co are distributed in plants to develop novel germplasms for the remediation of these heavy metals in contaminated soils. Here, an MTP gene that encodes the CDF (cation diffusion facilitator) protein in Populus trichocarpa, PtrMTP6, was screened as the key gene involved in the distribution of both Mn and Co in poplar. The PtrMTP6-GFP fusion protein was co-localized with the mRFP-VSR2, showing that PtrMTP6 proteins are present at the pre-vacuolar compartment (PVC). Yeast mutant complementation assays further identified that PtrMTP6 serves as a Mn and Co transporter, reducing yeast cell toxicity after exposure to excessive Mn or Co. Histochemical analyses showed that PtrMTP6 was mainly expressed in phloem, suggesting that PtrMTP6 probably involved in the Mn and Co transport via phloem in plants. Under excess Co, PtrMTP6 overexpressing poplar lines were more severely damaged than the control due to higher Co accumulations in young tissue. PtrMTP6 overexpressing lines showed little change in their tolerance to excess Mn, although young tissues also accumulated more Mn. PtrMTP6 play important roles in Mn and Co distribution in poplar and further research on its regulation will be important to increase bioremediation in Mn and Co polluted ecosystems.

Comparative and Systematic Omics Revealed Low Cd Accumulation of Potato StMTP9 in Yeast: Suggesting a New Mechanism for Heavy Metal Detoxification.

The metal tolerance protein (MTP) family is a very old family with evolutionary conservation and less specific amplification. It seems to retain the original functions of the ancestral genes and plays an important role in maintaining metal homeostasis in plant cells. We identified the potato MTP family members for the first time, the specific and conservative StMPTs were discovered by using systematic and comparative omics. To be surprised, members of the StMTP family seem to have mutated before the evolution of dicotyledon and monocotyledon, and even the loss of the entire subfamily (subfamily G6, G7). Interestingly, StMTP9 represents the conserved structure of the entire subfamily involved in toxic metal regulation. However, the gene structure and transmembrane domain of StMTP8 have undergone specific evolution, showing that the transmembrane domain (Motif13) located at the NH2 terminal has been replaced by the signal peptide domain, so it was selected as the control gene of StMTP9. Through real-time fluorescence quantitative analysis of StMTPs under Cd and Zn stress, a co-expression network was constructed, and it was found that StMTP9 responded significantly to Cd stress, while StMTP8 did the opposite. What excites us is that by introducing StMTPs 8/9 into the ∆ycf1 yeast cadmium-sensitive mutant strain, the functional complementation experiment proved that StMTPs 8/9 can restore Cd tolerance. In particular, StMTP9 can greatly reduce the cadmium content in yeast cells, while StMTP8 cannot. These findings provide a reference for further research on the molecular mechanism of potato toxic metal accumulation.

Genome-Wide Identification and Expression Analysis of Metal Tolerance Protein Gene Family in Medicago truncatula Under a Broad Range of Heavy Metal Stress.

Metal tolerance proteins (MTPs) encompass plant membrane divalent cation transporters to specifically participate in heavy metal stress resistance and mineral acquisition. However, the molecular behaviors and biological functions of this family in Medicago truncatula are scarcely known. A total of 12 potential MTP candidate genes in the M. truncatula genome were successfully identified and analyzed for a phylogenetic relationship, chromosomal distributions, gene structures, docking analysis, gene ontology, and previous gene expression. M. truncatula MTPs (MtMTPs) were further classified into three major cation diffusion facilitator (CDFs) groups: Mn-CDFs, Zn-CDFs, and Fe/Zn-CDFs. The structural analysis of MtMTPs displayed high gene similarity within the same group where all of them have cation_efflux domain or ZT_dimer. Cis-acting element analysis suggested that various abiotic stresses and phytohormones could induce the most MtMTP gene transcripts. Among all MTPs, PF16916 is the specific domain, whereas GLY, ILE, LEU, MET, ALA, SER, THR, VAL, ASN, and PHE amino acids were predicted to be the binding residues in the ligand-binding site of all these proteins. RNA-seq and gene ontology analysis revealed the significant role of MTP genes in the growth and development of M. truncatula. MtMTP genes displayed differential responses in plant leaves, stems, and roots under five divalent heavy metals (Cd2+, Co2+, Mn2+, Zn2+, and Fe2+). Ten, seven, and nine MtMTPs responded to at least one metal ion treatment in the leaves, stems, and roots, respectively. Additionally, MtMTP1.1, MtMTP1.2, and MtMTP4 exhibited the highest expression responses in most heavy metal treatments. Our results presented a standpoint on the evolution of MTPs in M. truncatula. Overall, our study provides a novel insight into the evolution of the MTP gene family in M. truncatula and paves the way for additional functional characterization of this gene family.

Molecular characterization and bioinformatics analysis of transporter genes associated with Cd-induced phytotoxicity in rice (Oryza sativa L.)

Cadmium (Cd) adversely affects the yield and quality of rice. It is, therefore, crucial to elucidate the consequences of Cd toxicity. Plant height, biomass, SPAD score, PSII efficiency, and photosynthetic performance index were all significantly reduced in Cd-stressed rice. Cd stress resulted in a simultaneous increase in Cd and Fe concentrations in both the roots and the shoots, accompanied by the significant upregulation of heavy metal ATPase (OsHMA2, OsHMA3), natural resistance-associated macrophage proteins (OsNramp1, OsNramp5), Fe-regulated transporters (OsIRT1), Fe-reductase oxidase (OsFRO1) genes, and FCR activity in roots. This implies that Cd uptake may be closely associated with Fe transporters resulted in physiological and photosynthetic damages in Cd-stressed rice. In silico analysis suggested that the localization of Cd-uptake proteins in the plasma membrane exhibiting transporter activity, among which two motifs were linked to the pfam_fs: Nramp domain. In a phylogenetic tree, HMA and Nramp genes were consistently positioned in the same cluster, while OsIRT1 and OsFRO1 were independently located. The key cis-acting elements were abscisic acid-responsiveness, methyl jasmonate-responsiveness, zein metabolism regulation, stress-responsiveness, salicylic acid-responsiveness, and gibberellin-responsiveness. An interactome map revealed the diverse functional partners of Cd-uptake genes, including MTP1 (metal tolerance protein 1), YSL6 (metal-nicotianamine transporter), IRO2 (Fe-regulated transcription factor 2), OsJ_16707 (a vacuolar Fe transporter homolog), YSL15 (an Fe-phytosiderophore transporter), and NAS2 (nicotianamine synthase), which were predominantly linked to Fe homeostasis. These findings greatly elucidate the Cd uptake mechanism in rice plants and can help to regulate Cd uptake either by breeding or silencing these transporters.

Genome assembly of the Chinese maize elite inbred line RP125 and its EMS mutant collection provide new resources for maize genetics research and crop improvement.

Maize is an important crop worldwide, as well as a valuable model with vast genetic diversity. Accurate genome and annotation information for a wide range of inbred lines would provide valuable resources for crop improvement and pan-genome characterization. In this study, we generated a high-quality de novo genome assembly (contig N50 of 15.43Mb) of the Chinese elite inbred line RP125 using Nanopore long-read sequencing and Hi-C scaffolding, which yield highly contiguous, chromosome-length scaffolds. Global comparison of the RP125 genome with those of B73, W22, and Mo17 revealed a large number of structural variations. To create new germplasm for maize research and crop improvement, we carried out an EMS mutagenesis screen on RP125. In total, we obtained 5818 independent M2 families, with 946 mutants showing heritable phenotypes. Taking advantage of the high-quality RP125 genome, we successfully cloned 10 mutants from the EMS library, including the novel kernel mutant qk1 (quekou: "missing a small part" in Chinese), which exhibited partial loss of endosperm and a starch accumulation defect. QK1 encodes a predicted metal tolerance protein, which is specifically required for Fe transport. Increased accumulation of Fe and reactive oxygen species as well as ferroptosis-like cell death were detected in qk1 endosperm. Our study provides the community with a high-quality genome sequence and a large collection of mutant germplasm.

Comprehensive genome wide identification and expression analysis of MTP gene family in tomato (Solanum lycopersicum) under multiple heavy metal stress.

Plant metal tolerance proteins (MTPs) play major roles in enhancing resistance to heavy metal tolerance and homeostasis. However, the role of MTPs genes in tomato, which is one of the most popular crops, is still largely limited. Hence, we investigated genome-wide study of tomato MTPs, including phylogenetic, duplication, gene structure, gene ontology and previous transcriptomic data analysis. Moreover, the MTPs expression behaviour under various heavy metals stress has rarely been investigated. In the current study, eleven MTP candidate genes were genome-wide identified and classified into three major groups; Mn-cation diffusion facilitators (CDFs), Fe/Zn-CDFs, and Zn-CDFs based on the phylogeny. Structural analysis of SlMTPs showed high gene similarity within the same group with cation_efflux or ZT_dimerdomains. Evolutionary analysis revealed that segmental duplication contributed to the expansion of the SlMTP family. Gene ontology further showed the vital roles of MTPs in metal-related processes. Tissue-specific expression profiling exhibited similar expression patterns in the same group, whereas gene expression varied among groups. The MTPs expression was evaluated after tomato treatments by five divalent heavy metals (Cd2+, Co2+, Mn2+, Zn2+, and Fe2+). SlMTP genes displayed differential responses in either plant leaves or roots under heavy metals treatments. Nine and ten SlMTPs responded to at least one metal ion treatment in leaves and roots, respectively. In addition SlMTP1, SlMTP3, SlMTP4, SlMTP8, SlMTP10 and SlMTP11 exhibited the highest expression responses in most of heavy metals treatments. Overall, our findings presented a standpoint on the evolution of MTPs and their evolution in tomato and paved the way for additional functional characterization under heavy metal toxicity.

Lateral roots but not root hairs contribute to high uptake of manganese and cadmium in rice.

Rice (Oryza sativa L.) can accumulate high manganese (Mn) in the shoots through uptake by the roots, which consist of crown roots, lateral roots and root hairs. We investigated the role of lateral roots and root hairs in Mn and cadmium (Cd) uptake by using two indica rice mutants defective in formation of lateral roots (osiaa11) and root hairs (osrhl1). The uptake of Mn and Cd in osiaa11 was significantly lower than that in wild type 'Kasalath', but there was no difference between wild type and osrhl1. Furthermore, a kinetic study showed that Mn uptake in osiaa11 was much lower than that in wild type and osrhl1 across a wide range of Mn concentrations. The role of lateral roots in Mn and Cd uptake was further confirmed in a japonica rice mutant defective in lateral root formation. We found that expression of Mn transporter gene Natural Resistance-Associated Macrophage Protein 5 (OsNRAMP5), but not of Metal Tolerance Protein 9 (OsMTP9), was lower in osiaa11 than in wild type; however, there were no differences between osrhl1 and the wild type. Immunostaining showed that OsNRAMP5 and OsMTP9 were localized in the exodermis and endodermis of crown roots and lateral roots, but not in the root hairs. Taken together, our results indicate that lateral roots, but not root hairs, play an important role in high Mn and Cd uptake in rice.

Genome-Wide Identification, Structure Characterization, Expression Pattern Profiling, and Substrate Specificity of the Metal Tolerance Protein Family in Canavalia rosea (Sw.) DC.

Plant metal tolerance proteins (MTPs) play key roles in heavy metal absorption and homeostasis in plants. By using genome-wide and phylogenetic approaches, the origin and diversification of MTPs from Canavalia rosea (Sw.) DC. was explored. Canavalia rosea (bay bean) is an extremophile halophyte with strong adaptability to seawater and drought and thereby shows specific metal tolerance with a potential phytoremediation ability. However, MTP genes in leguminous plants remain poorly understood. In our study, a total of 12 MTP genes were identified in C. rosea. Multiple sequence alignments showed that all CrMTP proteins possessed the conserved transmembrane domains (TM1 to TM6) and could be classified into three subfamilies: Zn-CDFs (five members), Fe/Zn-CDFs (five members), and Mn-CDFs (two members). Promoter cis-acting element analyses revealed that a distinct number and composition of heavy metal regulated elements and other stress-responsive elements existed in different promoter regions of CrMTPs. Analysis of transcriptome data revealed organ-specific expression of CrMTP genes and the involvement of this family in heavy metal stress responses and adaptation of C. rosea to extreme coral reef environments. Furthermore, the metal-specific activity of several functionally unknown CrMTPs was investigated in yeast. These results will contribute to uncovering the potential functions and molecular mechanisms of heavy metal absorption, translocation, and accumulation in C. rosea plants.

Characterization of the gene family encoding metal tolerance proteins in Triticum urartu: Phylogenetic, transcriptional, and functional analyses.

Homeostasis of microelements in organisms is vital for normal metabolism. In plants, the cation diffusion facilitator (CDF) protein family, also known as metal tolerance proteins (MTPs), play critical roles in maintaining trace metal homeostasis. However, little is known about these proteins in wheat. In this study, we characterized the MTP family of Triticum urartu, the donor of 'A' genome of Triticum aestivum, and analysed their phylogenetic relationships, sequence signatures, spatial expression patterns in the diploid wheat, and their transport activity when heterologously expressed. Nine MTPs were identified in the T. urartu genome database, and were classified and designated based on their sequence similarity to Arabidopsis thaliana (Arabidopsis) and Oryza sativa MTPs. Phylogenetic and sequence analyses indicated that the triticum urartu metal tolerance protein (TuMTP)s comprise three Zn-CDFs, two Fe/Zn-CDFs, and four Mn-CDFs; and can be further classified into six subgroups. Among the TuMTPs, there are no MTP2-5 and MTP9-10 counterparts but two MTP1/8/11 orthologs in relation to AtMTPs. It was also shown that members of the same cluster share similar sequence characteristic, i.e. number of introns, predicted transmembrane domains, and motifs. When expressed in yeast, TuMTP1 and TuMTP1.1 conferred tolerance to Zn and Co but not to other metal ions; while TuMTP8, TuMTP8.1, TuMTP11, and TuMTP11.1 conferred tolerance to Mn. When expressed in Arabidopsis, TuMTP1 localized to the tonoplast and significantly enhanced Zn and Co tolerance. TuMTPs showed diverse tissue-specific expression patterns. Taken together, the closely clustered TuMTPs share structural features and metal specificity but play diverse roles in the homeostasis of microelements in plant cells.

The Cd-induced morphological and photosynthetic disruption is related to the reduced Fe status and increased oxidative injuries in sugar beet

Cadmium (Cd) toxicity is a form of soil contamination that causes losses in plant growth and yield. Understanding the effects of Cd-induced changes in physiological and cellular processes will help scientists develop better scientific strategies for sugar beet plant improvement. Cd toxicity triggered a substantial decrease in morphological parameters and total soluble protein in sugar beets, as well as membrane damage and cell death. Furthermore, the SPAD score and photosynthetic OJIP parameters in leaves were severely affected due to Cd stress. This was correlated with the decreased FCR activity and BvIRT1 expression in roots, suggesting the adverse effect of Cd in Fe acquisition in sugar beet. Our findings also revealed that BvHMA3 and BvNRAMP3 were upregulated in Cd-exposed roots, indicating that these genes might be involved in Cd uptake in sugar beet. In silico analysis of BvHMA3 and BvNRAMP3 proteins showed close partnerships with several Arabidopsis genes mainly linked to metal tolerance protein, cation diffusion facilitator, vacuolar metal transporter, and vacuolar Fe transporter. Subsequently, Cd-exposed sugar beet showed severe sensitivity to oxidative damages resulted in elevated H2O2 and O2.- without possessed efficient antioxidant defense. Finally, growth retardation in Cd-exposed sugar beets is linked to photosynthetic inefficiency caused by low Fe levels and oxidative stress in cells. These results may be used to improve Cd-sensitive sugar beet plants by breeding or transgenic programs.

Changes in phenotype and gene expression under lead stress revealed key genetic responses to lead tolerance in Medicago sativa L.

Lead (Pb) is a serious heavy metal soil pollutant. It can be absorbed and accumulated by plant roots and impact plant growth. Medicago sativa L. (alfalfa) is a low-input forage and potential bioenergy crop, and improving its yield and quality has always been a focus of the alfalfa breeding industry. Little is known about the mechanism by which alfalfa responds to Pb stress at the molecular level. In this study, three alfalfa genotypes (a lead-resistant type (LR), a lead-sensitive type (LS) and an intermediate type (IN)) with contrasting abilities to resist lead were exposed to different durations of Pb treatment. Next-generation sequencing (NGS)-based RNA-seq technology was employed to characterize the root transcriptomes of three genotypes of alfalfa and identify differentially expressed genes (DEGs) during Pb stress. Genotypes LR and LS displayed different mechanisms of tolerance. In LR, the accumulation of more resistant substances was induced by the upregulation of sucrose synthase, glucan endo-1,3-beta-glucosidase, beta-amylase 3, probable trehalose-phosphate phosphatase J, 6-phosphofructo-2-kinase delta-1-pyrroline-5-carboxylate synthase (P5CS) and δ-ornithine aminotransferase (δ-OAT). In addition, flavin monooxygenase (YUCCA), 4-coumarate:CoA ligase-like protein (4CL), cinnamoyl-CoA reductase-like protein (CCR), ferulate 5-hydroxylase (F5H) and caffeic acid O-methyltransferase (COMT) were upregulated, leading to root development in a short time under Pb stress. Further study of the expression levels of metal transport-related genes, such as NRAMP (metal transporter), MATE (multidrug and toxin extrusion), HIPPs (heavy metal-associated isoprenylated plant proteins), MTP (metal tolerance protein), and ABC transporter, suggested that these genes were differentially expressed after lead treatment in the three alfalfa genotypes. Our research provides useful information for further studies on the molecular mechanism of Pb resistance in Medicago sativa L.

Transport, functions, and interaction of calcium and manganese in plant organellar compartments.

Calcium (Ca2+) and manganese (Mn2+) are essential elements for plants and have similar ionic radii and binding coordination. They are assigned specific functions within organelles, but share many transport mechanisms to cross organellar membranes. Despite their points of interaction, those elements are usually investigated and reviewed separately. This review takes them out of this isolation. It highlights our current mechanistic understanding and points to open questions of their functions, their transport, and their interplay in the endoplasmic reticulum (ER), vesicular compartments (Golgi apparatus, trans-Golgi network, pre-vacuolar compartment), vacuoles, chloroplasts, mitochondria, and peroxisomes. Complex processes demanding these cations, such as Mn2+-dependent glycosylation or systemic Ca2+ signaling, are covered in some detail if they have not been reviewed recently or if recent findings add to current models. The function of Ca2+ as signaling agent released from organelles into the cytosol and within the organelles themselves is a recurrent theme of this review, again keeping the interference by Mn2+ in mind. The involvement of organellar channels [e.g. glutamate receptor-likes (GLR), cyclic nucleotide-gated channels (CNGC), mitochondrial conductivity units (MCU), and two-pore channel1 (TPC1)], transporters (e.g. natural resistance-associated macrophage proteins (NRAMP), Ca2+ exchangers (CAX), metal tolerance proteins (MTP), and bivalent cation transporters (BICAT)], and pumps [autoinhibited Ca2+-ATPases (ACA) and ER Ca2+-ATPases (ECA)] in the import and export of organellar Ca2+ and Mn2+ is scrutinized, whereby current controversial issues are pointed out. Mechanisms in animals and yeast are taken into account where they may provide a blueprint for processes in plants, in particular, with respect to tunable molecular mechanisms of Ca2+ versus Mn2+ selectivity.

Computational characterization and expression profile of MTP1 gene associated with zinc homeostasis across dicot plant species

This study characterizes different MTP1 (metal tolerance protein)/ZAT (zinc transporter) homologs involved in zinc (Zn) homeostasis in plants. BLAST analysis of AtMTP1 protein against 15 plant species showed 21 MTP1 homologs. These MTP1 protein homologs generally contain ~400 residues, six transmembrane helices and cation transmembrane transporter activity (GO:0008324), which can be utilized in predicting Zn uptake and tolerance mechanisms. These MTP1 genes having 1 exon are located on chromosomes 2, 7, and 14. Motifs contain conserved sequences of 41–50 residues belonging to the cation efflux family, which may help target binding sites and transcription factors. Further, AtMTP1 shows close similarities with Glycine max and Medicago trunculata. Interactome analysis suggests the association of AtMTP1 with cadmium/zinc-transporting ATPase and ZIP metal ion transporter. The AtMTP1 network is predominantly connected to cadmium/zinc-transporting ATPase (heavy metal ATPase 2, HMA2; heavy metal ATPase 3, HMA3; heavy metal ATPase 4, HMA4), cation efflux protein (MTP11), and metal tolerance protein C3 (AT4G58060). The Genevestigator predicts the high expression potential of AtMTP1 in the apical root during senescence, seedling, and bolting stages in an association with 11 co-expressed genes, mainly linked to estradiol toxicity and heat stress. Besides, AtMTP1 protein homologs possess conserved N-glyco motifs and physicochemical properties. The similarity and interactions of AtMTP1 gene with other genes suggest that Zn homeostasis in plants is associated with the regulation of different genes. These findings may advance our understanding to further develop plants capable of maintaining Zn homeostasis under adverse conditions.

Lead Toxicity in Cereals: Mechanistic Insight Into Toxicity, Mode of Action, and Management.

Cereals are the major contributors to global food supply, accounting for more than half of the total human calorie requirements. Sustainable availability of quality cereal grains is an important step to address the high-priority issue of food security. High concentrations of heavy metals specifically lead (Pb) in the soil negatively affect biochemical and physiological processes regulating grain quality in cereals. The dietary intake of Pb more than desirable quantity via food chain is a major concern for humans, as it can predispose individuals to chronic health issues. In plant systems, high Pb concentrations can disrupt several key metabolic processes such as electron transport chain, cellular organelles integrity, membrane stability index, PSII connectivity, mineral metabolism, oxygen-evolving complex, and enzymatic activity. Plant growth-promoting rhizobacteria (PGPR) has been recommended as an inexpensive strategy for remediating Pb-contaminated soils. A diverse group of Ascomycetes fungi, i.e., dark septate endophytes is successfully used for this purpose. A symbiotic relationship between endophytes and host cereal induces Pb tolerance by immobilizing Pb ions. Molecular and cellular modifications in plants under Pb-stressed environments are explained by transcription factor families such as bZIP, ERF, and GARP as a regulator. The role of metal tolerance protein (MTP), natural resistance-associated macrophage protein (NRAMP), and heavy metal ATPase in decreasing Pb toxicity is well known. In the present review, we provided the contemporary synthesis of existing data regarding the effects of Pb toxicity on morpho-physiological and biochemical responses of major cereal crops. We also highlighted the mechanism/s of Pb uptake and translocation in plants, critically discussed the possible management strategies and way forward to overcome the menace of Pb toxicity in cereals.

Comparative transcriptome profile analysis of rice varieties with different tolerance to zinc deficiency

Zinc (Zn) is an indispensable element for rice growth. Zn deficiency results in brown blotches and streaks 2-3 weeks after transplanting, as well as stunting, reduced tillering, and low productivity of rice plants. These processes are controlled by different families of expressed genes. A comparative transcriptome profile analysis was conducted using the roots of two Zn deficiency tolerant varieties (UCP122 and KALIBORO26) and two sensitive varieties (IR26 and IR64) by merging data from untreated control (CK) and Zn deficiency treated samples. Results revealed a total of 4,688 differentially expressed genes (DEGs) between the normal Zn and deficient conditions, with 2,702 and 1,489 unique DEGs upregulated and downregulated, respectively. Functional enrichment analysis identified transcription factors (TFs), such as WRKY, MYB, ERF, and bHLH which are important in the regulation of the Zn deficiency response. Furthermore, chitinases, jasmonic acid, and phenylpropanoid pathways were found to be important in the Zn deficiency response. The metal tolerance protein (MTP) genes also appeared to play an important role in conferring tolerance to Zn deficiency. A heavy metal-associated domain-containing protein 7 was associated with tolerance to Zn deficiency and negatively regulated downstream genes. Collectively, our findings provide valuable expression patterns and candidate genes for the study of molecular mechanisms underlying the response to Zn deficiency and for improvements in breeding for tolerance to Zn deficiency in rice.

Скрининг растений-регенерантов сахарной свёклы на наличие гена устойчивости к тяжелым металлам MTP4

The MTP4 (Metal Tolerance Protein 4) gene, which controls the resistance of sugar beet plants to heavy metals and is located on the 3rd chromosome has been studied. Micro clones grown in vitro with lethal Cd doses were used as explants. For PCR, specific primers MTP4A F/R and MTP4B F/R, covering the 2nd, 3rd and 4th exons and introns between them in the MTP4 gene, were used. Samples with resistance to heavy metals (in particular, Cd) contained some nucleotide changes in the 4th exon resulting in amino acid substitutions in the polypeptide chain. Based on the results of the bioinformatics analysis, it can be assumed with a high probability that the resistance of the studied breeding materials to heavy metals arose due to the presence of certain polymorphisms in the analyzed sugar beet gene. Geneious Prime software as a tool for the MTP4 gene sequence analysis will probably allow the in vitro selection of resistant forms for the subsequent use in the breeding scheme. Key words: sugar beet, МТР genes, genetic polymorphism, PCR, molecular genetic markers, breeding, DNA

Expression of a Brassica napus metal transport protein (BnMTP3) in Arabidopsis thaliana confers tolerance to Zn and Mn

The essential micronutrient elements zinc (Zn) and manganese (Mn) are crucial for plant growth and development. As an important oil crop, the yield and quality of rapeseed are affected by Zn and Mn toxicity. The cation diffusion facilitator (CDF) family of proteins play significant roles in maintaining intracellular ionic homeostasis and tolerance in plants. However, research on CDF proteins in rapeseed is lacking. In this study, the function of a Brassica napus cation diffusion facilitator/ metal tolerance protein (CDF/MTP) was investigated. The protein, abbreviated BnMTP3 is homologous to the Arabidopsis thaliana MTP3 (AtMTP3). Heterologous expression of BnMTP3 in yeast enhanced tolerance and intracellular sequestration of Zn and Mn. Expression of BnMTP3 in A. thaliana increased Zn and Mn tolerance and markedly increased Zn accumulation in roots. Quantitative RT-PCR analysis showed that BnMTP3 is primarily expressed in roots, and subcellular localization suggested that BnMTP3 is localized in the trans-Golgi network (TGN) and the prevacuolar compartment (PVC) in Arabidopsis and rape protoplast. After treatment with Zn and Mn, BnMTP3 was observed on the vacuolar membrane in transgenic Arabidopsis lines. These findings suggest that BnMTP3 confers Zn and Mn tolerance by sequestering Zn and/or Mn into the vacuole.