wri1转录因子调控拟南芥盐和重金属非生物胁迫相关的文献

WRINKLED1 (WRI1) has targets other than fatty acid biosynthesis genes that mediate the growth of roots.

Members of the highly conserved class of BEACH domain containing proteins (BDCPs) have been established as broad facilitators of protein–protein interactions and membrane dynamics in the context of human diseases like albinism, bleeding diathesis, impaired cellular immunity, cancer predisposition, and neurological dysfunctions. Also, the Arabidopsis thaliana BDCP SPIRRIG (SPI) is important for membrane integrity, as spi mutants exhibit split vacuoles. In this work, we report a novel molecular function of the BDCP SPI in ribonucleoprotein particle formation. We show that SPI interacts with the P-body core component DECAPPING PROTEIN 1 (DCP1), associates to mRNA processing bodies (P-bodies), and regulates their assembly upon salt stress. The finding that spi mutants exhibit salt hypersensitivity suggests that the local function of SPI at P-bodies is of biological relevance. Transcriptome-wide analysis revealed qualitative differences in the salt stress-regulated transcriptional response of Col-0 and spi. We show that SPI regulates the salt stress-dependent post-transcriptional stabilization, cytoplasmic agglomeration, and localization to P-bodies of a subset of salt stress-regulated mRNAs. Finally, we show that the PH-BEACH domains of SPI and its human homolog FAN (Factor Associated with Neutral sphingomyelinase activation) interact with DCP1 isoforms from plants, mammals, and yeast, suggesting the evolutionary conservation of an association of BDCPs and P-bodies.

No abstract available

N6 -methyladenosine (m6 A) is an mRNA modification widely found in eukaryotes and plays a crucial role in plant development and stress responses. FIONA1 (FIO1) is a recently identified m6 A methyltransferase that regulates Arabidopsis (Arabidopsis thaliana) floral transition; however, its role in stress response remains unknown. In this study, we demonstrate that FIO1-mediated m6 A methylation plays a vital role in salt stress response in Arabidopsis. The loss-of-function fio1 mutant was sensitive to salt stress. Importantly, the complementation lines expressing the wild-type FIO1 exhibited the wild-type phenotype, whereas the complementation lines expressing the mutant FIO1m , in which two critical amino acid residues essential for methyltransferase activity were mutated, did not recover the wild-type phenotype under salt stress, indicating that the salt sensitivity is associated with FIO1 methyltransferase activity. Furthermore, FIO1-mediated m6 A methylation regulated ROS production and affected the transcript level of several salt stress-responsive genes via modulating their mRNA stability in an m6 A-dependent manner in response to salt stress. Importantly, FIO1 is associated with salt stress response by specifically targeting and differentially modulating several salt stress-responsive genes compared with other m6 A writer. Collectively, our findings highlight the molecular mechanism of FIO1-mediated m6 A methylation in the salt stress adaptation.

No abstract available

Developmental plasticity is critical for plants to adapt to constantly changing environments. Plant root hairs display dramatic plasticity under different environments and therefore play crucial roles in defense against environmental stressors. Here, we report the isolation of an Arabidopsis mutant, salinityover-sensitivemutant 1–1 (som1-1), also exhibiting root hair developmental defects. Map-based cloning and allelic analyses confirmed that som1-1 is a new mutant allele of SPIRRIG (SPI), which encodes a Beige and Chediak Higashi (BEACH) domain-containing protein. SPI has been reported to facilitate actin dependent root hair development by temporally and spatially regulating the expression of BRICK1 (BRK1), a subunit of the SCAR/WAVE actin nucleating promoting complex. Our living cell imaging examinations revealed that salt stress induces an altered actin organization in root hair that mimics those in the spi mutant, implying SPI may respond to salt stress induced root hair plasticity by modulating actin cytoskeleton organization. Furthermore, we found BRK1 is also involved in root hair developmental change under salt stress, and overexpression of BRK1 resulted in root hairs over-sensitive to salt stress as those in spi mutant. Moreover, based on biochemical analyses, we found BRK1 is unstable and SPI mediates BRK1 stability. Functional loss of SPI results in the accumulation of steady-state of BRK1.

The maintenance of sodium/potassium (Na+/K+) homeostasis in plant cells is essential for salt tolerance. Plants export excess Na+ out of cells mainly through the Salt Overly Sensitive (SOS) pathway, activated by a calcium signal; however, it is unknown whether other signals regulate the SOS pathway and how K+ uptake is regulated under salt stress. Phosphatidic acid (PA) is emerging as a lipid signaling molecule that modulates cellular processes in development and the response to stimuli. Here, we show that PA binds to the residue Lys57 in SOS2, a core member of the SOS pathway, under salt stress, promoting the activity and plasma membrane localization of SOS2, which activates the Na+/H+ antiporter SOS1 to promote the Na+ efflux. In addition, we reveal that PA promotes the phosphorylation of SOS3‐like calcium‐binding protein 8 (SCaBP8) by SOS2 under salt stress, which attenuates the SCaBP8‐mediated inhibition of Arabidopsis K+ transporter 1 (AKT1), an inward‐rectifying K+ channel. These findings suggest that PA regulates the SOS pathway and AKT1 activity under salt stress, promoting Na+ efflux and K+ influx to maintain Na+/K+ homeostasis.

No abstract available

SALT OVERLY SENSITIVE1 (SOS1) is a key component of plant salt tolerance. However, how SOS1 transcription is dynamically regulated in plant response to different salinity conditions remains elusive. Here, we report that C-type Cyclin1; 1 (CycC1; 1) negatively regulates salt tolerance by interfering with WRKY75-mediated transcriptional activation of SOS1 in Arabidopsis (Arabidopsis thaliana). Disruption of CycC1; 1 promotes SOS1 expression and salt tolerance in Arabidopsis because CycC1; 1 interferes with RNA polymerase II recruitment by occupying the SOS1 promoter. Enhanced salt tolerance of the cycc1; 1 mutant was completely compromised by an SOS1 mutation. Moreover, CycC1; 1 physically interacts with the transcription factor WRKY75, which can bind to the SOS1 promoter and activate SOS1 expression. In contrast to the cycc1; 1 mutant, the wrky75 mutant has attenuated SOS1 expression and salt tolerance, whereas overexpression of SOS1 rescues the salt sensitivity of wrky75. Intriguingly, CycC1; 1 inhibits WRKY75-mediated transcriptional activation of SOS1 via their interaction. Thus, increased SOS1 expression and salt tolerance in cycc1; 1 was abolished by WRKY75 mutation. Our findings demonstrate that CycC1; 1 forms a complex with WRKY75 to inactivate SOS1 transcription under low salinity conditions. By contrast, under high salinity conditions, SOS1 transcription and plant salt tolerance is activated at least partially by increased WRKY75 expression but decreased CycC1; 1 expression.

No abstract available

Significance Plants mobilize the endomembrane system for restoring homeostasis when stimulated by stresses. Though the function of ufmylation in ER stress and autophagy has been described in mammals, their detailed roles in plant development and stress responses remain elusive. In this study, we identified Ufl1 as an ATG8 interactor in Arabidopsis and demonstrated that Ufl1 maintains ER homeostasis upon salt stress via regulating ER-phagy by mechanistic connection with the autophagy-related (ATG) machinery. Our study thus sheds light on the pleiotropic functions of the ufmylation cascade in plants under abiotic stresses and underpins the cross-talk among ER stress, salt stress, and autophagy.

Abiotic stresses such as salinity and low temperature have serious impact on peanut growth and yield. The present work investigated the function of a MYB-related transcription factor gene AhMYB30 obtained from peanut under salt and low temperature stresses by transgenic methods. The results indicated that the overexpression of AhMYB30 in Arabidopsis could enhance the resistance of transgenic plants to freezing and salt stresses. The expression of stress-response genes RD29A (Response-to-Dehydration 29A), COR15A (Cold-Regulated 15A), KIN1 (Kinesin 1) and ABI2 (Abscisic acid Insensitive 2) increased in transgenic plants compared with in wild-type. Subcellular localization and transcriptional autoactivation validation demonstrated that AhMYB30 has essential features of transcription factors. Therefore, AhMYB30 may increase salt and freezing stress tolerance as the transcription factor (TF) in Arabidopsis through both DREB/CBF and ABA-signaling pathways. Our results lay the theoretical foundation for exploring stress resistance mechanisms of peanut and offering novel genetic resources for molecular breeding.

Post-translational modifications (PTMs) of proteins play important roles in the acclimation of plants to environmental stress. Lysine acetylation is a dynamic and reversible PTM, which can be removed by histone deacetylases. Here we investigated the role of lysine acetylation in the response of Arabidopsis leaves to one week of salt stress. A quantitative mass spectrometry analysis revealed an increase in lysine acetylation of several proteins from cytosol and plastids, which was accompanied by altered histone deacetylase activities in the salt-treated leaves. While activities of HDA14 and HDA15 were decreased upon salt stress, HDA5 showed a mild and HDA19 a strongly increased activity. Since HDA5 is a cytosolic-nuclear enzyme from the class II histone deacetylase family with yet unknown protein substrates, we performed a lysine acetylome analysis on hda5 mutants and characterized its substrate proteins. Next to histone H2B, the salt stress responsive transcription factor GT2L and the dehydration-related protein ERD7 were identified as HDA5 substrates. In addition, in protein-protein interaction studies, HDA18 was discovered, among other interacting proteins, to work in a complex together with HDA5. Altogether this study revealed the substrate proteins of HDA5 and identified new lysine acetylation sites which are hyperacetylated upon salt stress. The identification of the specific histone deacetylase substrate proteins, apart from histones, will be important to unravel the acclimation response of Arabidopsis to salt stress and their role in plant physiology.

No abstract available

Excessive Na+ in soils inhibits plant growth. Here, we report that Na+ stress triggers primary calcium signals specifically in a cell group within the root differentiation zone, thus forming a "sodium-sensing niche" in Arabidopsis. The amplitude of this primary calcium signal and the speed of the resulting Ca2+ wave dose-dependently increase with rising Na+ concentrations, thus providing quantitative information about the stress intensity encountered. We also delineate a Ca2+-sensing mechanism that measures the stress intensity in order to mount appropriate salt detoxification responses. This is mediated by a Ca2+-sensor-switch mechanism, in which the sensors SOS3/CBL4 and CBL8 are activated by distinct Ca2+-signal amplitudes. Although the SOS3/CBL4-SOS2/CIPK24-SOS1 axis confers basal salt tolerance, the CBL8-SOS2/CIPK24-SOS1 module becomes additionally activated only in response to severe salt stress. Thus, Ca2+-mediated translation of Na+ stress intensity into SOS1 Na+/H+ antiporter activity facilitates fine tuning of the sodium extrusion capacity for optimized salt-stress tolerance.

MYB (v-MYB avian myeloblastosis viral oncogene homolog) transcription factor (TF) family has numerous members with complex and diverse functions, which perform an integral role in regulating the plant’s response to adversity. This study used cloning to obtain a novel MYB TF gene from the diploid strawberry Fragaria vesca, which was given the designation FvMYB44. Subcellular localization results showed that the protein of FvMYB44 was a nuclear localization protein. The resistance of Arabidopsis thaliana to salt and low temperature stresses was greatly enhanced by the overexpression of FvMYB44. When subjected to salt and temperature stress, transgenic plants showed higher proline and chlorophyll concentrations and higher superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities than wild-type (WT) and unloaded line (UL) of A. thaliana. In contrast, WT and UL lines had higher malondialdehyde (MDA) content and reactive oxygen species ROS (O2− and H2O2) content. These findings suggest that FvMYB44 may perform a role in controlling the response of A. thaliana to cold and salt stress.

The grape (Vitis vinifera L.) not only has a long history of cultivation, but also has rich nutritional value and high economic value. However, grapes often face many threats in the growth process. For example, low temperature and salt stress restrict the growth status, yield, and geographical distribution of grapes. WRKY, as one of the largest transcription factor (TF) families in plants, participates in the response of plants to stress. VvWRKY28, a new zinc finger type transcriptional regulator gene, was isolated from Beichun (V. vinifera × V.amurensis) in this study. From the subcellular localization results, it can be concluded that VvWRKY28 was localized in the nucleus. The expression of VvWRKY28 was enriched in leaves (young and mature leaves), and cold and high salt conditions can induce high expression of VvWRKY28. After being transferred into Arabidopsis, VvWRKY28 greatly improved the tolerance of Arabidopsis to low temperature and high salt and also changed many physiological and biochemical indicators of transgenic Arabidopsis to cope with cold and high salt stimulation. The content of malondialdehyde (MDA) was decreased, but for chlorophyll and proline, their content increased, and the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were improved. In addition, under cold stress, binding with cis-acting elements promotes the expression of downstream genes related to cold stress (RAB18, COR15A, ERD10, PIF4, COR47, and ICS1). Moreover, it also plays an active role in regulating the expression of genes related to salt stress (NCED3, SnRK2.4, CAT2, SOD1, SOS2, and P5CS1) under salt stress. Therefore, these results provide evidence that VvWRKY28 may play a role in the process of plant cold and salt stress tolerance.

Integration of light signaling and diverse abiotic stress responses contribute to plant survival in a changing environment. A number of reports have indicated that light signals contribute a plant's ability to deal with heat, cold, and stress. However, the molecular link between light signaling and the salt-response pathways remains unclear. We demonstrate here that increasing light intensity elevates the salt stress tolerance of plants. Depletion of HY5, a key component of light signaling, causes Arabidopsis thaliana to become salinity sensitive. Interestingly, the small heat shock protein (sHsp) family genes are up-regulated in hy5-215 mutant plants, and HsfA2 is commonly involved in the regulation of these sHsps. We found that HY5 directly binds to the G-box motifs in the HsfA2 promoter, with the cooperation of HISTONE DEACETYLASE 9 (HDA9), to repress its expression. Furthermore, the accumulation of HDA9 and the interaction between HY5 and HDA9 are significantly enhanced by salt stress. On the contrary, high temperature triggers HY5 and HDA9 degradation, which leads to dissociation of HY5-HDA9 from the HsfA2 promoter, thereby reducing salt tolerance. Under salt and heat stress conditions, fine-tuning of protein accumulation and an interaction between HY5 and HDA9 regulate HsfA2 expression. This implies that HY5, HDA9, and HsfA2 play important roles in the integration of light signaling with salt stress and heat shock response. This article is protected by copyright. All rights reserved.

Zinc finger proteins are widely involved and play an important role in plant growth and abiotic stress. In this research, MdZAT5, a gene encoding C2H2-type zinc finger protein, was cloned and investigated. The MdZAT5 was highly expressed in flower tissues by qRT-PCR analyses and GUS staining. Promoter analysis showed that MdZAT5 contained multiple response elements, and the expression levels of MdZAT5 were induced by various abiotic stress treatments. Overexpression of MdZAT5 in apple calli positively regulated anthocyanin accumulation by activating the expressions of anthocyanin biosynthesis-related genes. Overexpression of MdZAT5 in Arabidopsis also enhanced the accumulation of anthocyanin. In addition, MdZAT5 increased the sensitivity to salt stress in apple calli. Ectopic expression of MdZAT5 in Arabidopsis reduced the expression of salt-stress-related genes (AtNHX1 and AtABI1) and improved the sensitivity to salt stress. In conclusion, these results suggest that MdZAT5 plays a positive regulatory role in anthocyanin accumulation and negatively regulates salt resistance.

As the most abundant internal modification in mRNA, N6 -methyladenosine (m6 A) methylation of RNA is emerging as a new layer of epitranscriptomic gene regulation in cellular processes, including embryo development, flowering time control, microspores generation, and fruit-ripening, in plants. However, the cellular role of m6 A in plant response to environmental stimuli remains largely unexplored. In this study, we show that m6 A methylation plays an important role in salt stress tolerance in Arabidopsis. All mutants of m6 A writer components, including MTA, MTB, VIRILIZER (VIR), and HAKAI, displayed salt-sensitive phenotypes in an m6 A-dependent manner. The vir mutant, in which the m6 A level was most highly reduced, exhibited salt-hypersensitive phenotypes. Analysis of m6 A methylome in the vir mutant revealed a transcriptome-wide loss of m6 A modification in the 3'UTR. We further demonstrated that VIR-mediated m6 A methylation modulates reactive oxygen species homeostasis by negatively regulating the mRNA stability of several salt stress negative regulators, including ATAF1, GI, and GSTU17, via affecting 3'UTR lengthening linked to alternative polyadenylation. Our results highlight the important role played by epitranscriptomic mRNA methylation in the salt stress response of Arabidopsis and indicate a strong link between m6 A methylation and 3'UTR length and mRNA stability during stress adaptation.

No abstract available

Abstract The precise timing of flowering in adverse environments is critical for plants to secure reproductive success. We report a mechanism in Arabidopsis (Arabidopsis thaliana) controlling the time of flowering by which the S-acylation-dependent nuclear import of the protein SALT OVERLY SENSITIVE3/CALCINEURIN B-LIKE4 (SOS3/CBL4), a Ca2+-signaling intermediary in the plant response to salinity, results in the selective stabilization of the flowering time regulator GIGANTEA inside the nucleus under salt stress, while degradation of GIGANTEA in the cytosol releases the protein kinase SOS2 to achieve salt tolerance. S-acylation of SOS3 was critical for its nuclear localization and the promotion of flowering, but partly dispensable for salt tolerance. SOS3 interacted with the photoperiodic flowering components GIGANTEA and FLAVIN-BINDING, KELCH REPEAT, F-BOX1 and participated in the transcriptional complex that regulates CONSTANS to sustain the transcription of CO and FLOWERING LOCUS T under salinity. Thus, the SOS3 protein acts as a Ca2+- and S-acylation-dependent versatile regulator that fine-tunes flowering time in a saline environment through the shared spatial separation and selective stabilization of GIGANTEA, thereby connecting two signaling networks to co-regulate the stress response and the time of flowering.

MYB transcription factors (TFs) mediate plant responses and defenses to biotic and abiotic stresses. The effects of overexpression of MYB37, an R2R3 MYB subgroup 14 transcription factors in Arabidopsis thaliana, on chlorophyll content, chlorophyll fluorescence parameters, reactive oxygen species (ROS) metabolism, and the contents of osmotic regulatory substances were studied under 100 mM NaCl stress. Compared with the wild type (Col-0), MYB37 overexpression significantly alleviated the salt stress symptoms in A. thaliana plants. Chlorophyll a (Chl a) and chlorophyll b (Chl b) contents were significantly decreased in OE-1 and OE-2 than in Col-0. Particularly, the Chl a/b ratio was also higher in OE-1 and OE-2 than in Col-0 under NaCl stress. However, MYB37 overexpression alleviated the degradation of chlorophyll, especially Chl a. Salt stress inhibited the activities of PSII and PSI in Arabidopsis leaves, but did not affect the activity of PSII electron donor side oxygen-evolving complex (OEC). MYB37 overexpression increased photosynthesis in Arabidopsis by increasing PSII and PSI activities. MYB37 overexpression also promoted the transfer of electrons from QA to QB on the PSII receptor side of Arabidopsis under NaCl stress. Additionally, MYB37 overexpression increased Y(II) and Y(NPQ) of Arabidopsis under NaCl stress and decreased Y(NO). These results indicate that MYB37 overexpression increases PSII activity and regulates the proportion of energy dissipation in Arabidopsis leaves under NaCl stress, thus decreasing the proportion of inactivated reaction centers. Salt stress causes excess electrons and energy in the photosynthetic electron transport chain of Arabidopsis leaves, resulting in the release of reactive oxygen species (ROS), such as superoxide anion and hydrogen peroxide, leading to oxidative damage. Nevertheless, MYB37 overexpression reduced accumulation of malondialdehyde in Arabidopsis leaves under NaCl stress and alleviated the degree of membrane lipid peroxidation caused by ROS. Salt stress also enhanced the accumulation of soluble sugar (SS) and proline (Pro) in Arabidopsis leaves, thus reducing salt stress damage to plants. Salt stress also degraded soluble protein (SP). Furthermore, the accumulation of osmoregulation substances SS and Pro in OE-1 and OE-2 was not different from that in Col-0 since MYB37 overexpression in Arabidopsis OE-1, and OE-2 did not significantly affect plants under NaCl stress. However, SP content was significantly higher in OE-1 and OE-2 than in Col-0. These results indicate that MYB37 overexpression can alleviate the degradation of Arabidopsis proteins under NaCl stress, promote plant growth and improve salt tolerance.

AtUSP17 is a multiple stress-inducible gene that encodes a universal stress protein (USP) in Arabidopsis thaliana. In the present study, we functionally characterized AtUSP17 using its knock-down mutant, Atusp17, and AtUSP17-overexpression lines (WTOE). The overexpression of AtUSP17 in wild-type and Atusp17 mutant Arabidopsis plants resulted in higher sensitivity to salt stress during seed germination than WT and Atusp17 mutant lines. In addition, the WTOE and FC lines exhibited higher abscisic acid (ABA) sensitivity than Atusp17 mutant during germination. The exogenous application of ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) was able to rescue the salt hypersensitive phenotype of WTOE lines. In contrast, AgNO3 , an ethylene action inhibitor, further blocked the effect of ACC during germination. The addition of ACC under salt stress resulted in reduced reactive oxygen species (ROS) accumulation, expression of ABA-responsive genes, improved proline synthesis, increased expression of positive regulators of ethylene signaling and antioxidant defense genes with enhanced antioxidant enzyme activities. The WTOE lines exhibited salt sensitivity even at the adult plant stage, while Atusp17 mutant exhibited higher salt tolerance with higher chlorophyll, relative water content and lower electrolyte leakage as compared to WT. The BAR interaction viewer database and available literature mining identified AtUSP17-interacting proteins, which include RGS1, RACK1C and PRN1 involved in G-protein signaling, which play a crucial role in salt stress responses. Based on the present study and available literature, we proposed a model in which AtUSP17 negatively mediates salt tolerance in Arabidopsis through modulation of ethylene, ABA, ROS and G-protein signaling and responses. This article is protected by copyright. All rights reserved.

Heat shock transcription factors (HSF) are divided into classes A, B and C. Class A transcription factors are generally recognized as transcriptional activators, while functional characterization of class B and C heat shock transcription factors have not been fully developed in most plant species. We isolated and characterized a novel HSF transcription factor gene, TrHSFB2a (a class B HSF) gene, from the drought stress-sensitive forage crop species, white clover (Trifolium repens). TrHSFB2a was highly homologous to MtHSFB2b, CarHSFB2a, AtHSFB2b and AtHSFB2a. The expression of TrHSFB2a was strongly induced by drought (PEG6000 15% w/v), high temperature (35 °C) and salt stresses (200 mM L−1 NaCl) in white clover, while subcellular localization analysis showed that it is a nuclear protein. Overexpression of the white clover gene TrHSFB2a in Arabidopsis significantly reduced fresh and dry weight, relative water contents (RWC), maximum photosynthesis efficiency (Fv/Fm) and performance index on the absorption basis (PIABS), while it promoted leaf senescence, relative electrical conductivity (REC) and the contents of malondialdehyde (MDA) compared to a wild type under drought, heat and salt stress conditions of Arabidopsis plants. The silencing of its native homolog (AtHSFB2a) by RNA interference in Arabidopsis thaliana showed opposite trends by significantly increasing fresh and dry weights, RWC, maximum photosynthesis efficiency (Fv/Fm) and performance index on the absorption basis (PIABS) and reducing REC and MDA contents under drought, heat and salt stress conditions compared to wild type Arabidopsis plants. These phenotypic and physiological indicators suggested that the TrHSFB2a of white clover functions as a negative regulator of heat, salt and drought tolerance. The bioinformatics analysis showed that TrHSFB2a contained the core B3 repression domain (BRD) that has been reported as a repressor activator domain in other plant species that might repress the activation of the heat shock-inducible genes required in the stress tolerance process in plants. The present study explores one of the potential causes of drought and heat sensitivity in white clover that can be overcome to some extent by silencing the TrHSFB2a gene in white clover.

Brassinosteroids (BRs) play critical roles in plant growth and development, as well as in responses to abiotic stresses. The BRASSINAZOLE RESISTANT 1 (BZR1) and BRI1-EMS-SUPPRESSOR 1 (BES1) families of transcription factors have been elucidated largely in Arabidopsis and rice but not in other plant species. Here, we studied the functional characterization of a tomato (Solanum lycopersicum) BZR homolog gene, SlBZR1, in BR-regulated plant growth and tolerance to salt stress. SlBZR1 was highly expressed in the flowers and developing fruits of tomato. Both SlBZR1 and SlBZR1D (proline to leucine mutation at the 239th amino acid of SlBZR1) were transcriptional repressors and localized in the nucleus. SlBZR1 or SlBZR1D could interact with SlMYB30, SlMYBL2, SlPIF4, SlHAT1, SlIWS1 and SlREF6 in tomato. Overexpression of SlBZR1D enhanced the BR response and improved tolerance to salt stress in Arabidopsis, consistent with the phenotype of the Arabidopsis bes1-D mutant. Moreover, SlBZR1D-overexpressing tomato lines showed a short plant height, smaller and curly leaves, and delayed flowering. Additionally, SlBZR1D positively regulated salt tolerance in tomato and upregulated the expression of multiple stress-related genes. Our study provides new insights for understanding the function and mechanism of BZR transcription factors in BR-regulated plant growth and abiotic stress responses.

Alternative splicing (AS) is emerging as a critical co-transcriptional regulation for plants in response to environmental stresses. Although multiple splicing factors have been linked to the salt-sensitive signaling network, the molecular mechanism remains unclear. We discovered that a conserved SR-like protein, SR45a, as a component of the spliceosome, was involved in post-transcriptional regulation of salinity tolerance in Arabidopsis thaliana. Furthermore, SR45a was required for the AS and mRNA maturation of several salt-tolerance genes. Two alternatively spliced variants of SR45a were induced by salt stress, full-length SR45a-1a and the truncated isoform SR45a-1b, respectively. Lines with overexpression of SR45a-1a and SR45a-1b exhibited hypersensitive to salt stress. Our data indicated that SR45a directly interacted with the cap-binding complex (CBC) subunit cap-binding protein 20 (CBP20) which mediated salt stress responses. Instead of binding to other spliceosome components, SR45a-1b promoted the association of SR45a-1a with CBP20, therefore mediating salt stress signal transduction pathways. Additionally, the mutations in SR45a and CBP20 led to different salt stress phenotypes. Together, these results provide the evidence that SR45a-CBP20 acts as a regulatory complex to regulate the plant response to salt stress, through a regulatory mechanism to fine-tune the splicing factors, especially in stressful conditions.

Salt is one of the most common abiotic stresses, causing ionic and osmotic pressure changes that affect plant growth and development. In this work, we present molecular and genetic evidence that Arabidopsis Toxicos en Levadura 12 (ATL12) is involved in both salt stress and in the abscisic acid response to this stress. We demonstrate that ATL12 is highly induced in response to salt stress and that atl12 mutants have a lower germination rate, decreased root length, and lower survival rate compared to the Col-0 wild-type in response to salt stress. Overexpression of ATL12 increases expression of the salt stress-associated genes SOS1/2, and ABA-responsive gene RD29B. Additionally, higher levels of reactive oxygen species are detected when ATL12 is overexpressed, and qRT-PCR showed that ATL12 is involved in the AtRBOHD/F-mediated signaling. ATL12 expression is also highly induced by ABA treatment. Mutants of atl12 are hypersensitive to ABA and have a shorter root length. A decrease in water loss and reduced stomatal aperture were also observed in atl12 mutants in response to ABA. ABA-responsive genes RD29B and RAB18 were downregulated in atl12 mutants but were upregulated in the overexpression line of ATL12 in response to ABA. Taken together our results suggest that ATL12 modulates the response to salt stress and is involved in the ABA signaling pathway in Arabidopsis thaliana.

Sessile plants constantly experience environmental stresses in nature. They must have evolved effective mechanisms to balance growth with stress response. Here we report the MADS-box transcription factor AGL16 acting as a negative regulator in stress response in Arabidopsis. Loss-of-AGL16 confers resistance to salt stress in seed germination, root elongation, and soil-grown plants, while elevated AGL16 expression confers the opposite phenotypes compared with wild type. However, the sensitivity to ABA in seed germination is inversely correlated with AGL16 expression level. Transcriptomic comparison revealed that the improved salt resistance of agl16 mutants was largely attributed to enhanced expression of stress responsive transcriptional factors and the genes involved in ABA signaling and ion homeostasis. We further demonstrated that AGL16 directly binds to the CArG motifs in the promoter of HKT1;1, HsfA6a, and MYB102 and represses their expression. Genetic analyses with double mutants also support that HsfA6a and MYB102 are target genes of AGL16. Taken together, our results show that AGL16 acts as a negative regulator transcriptionally suppressing key components in stress response and may play a role in balancing stress response with growth.

Plant metabolites are dynamically modified and distributed under environmental changes. However, it is poorly understood how metabolites change functions in plant stress responses. Maintaining ion homeostasis under salt stress requires coordinately activating two type central regulators: PM H+-ATPase and Na+/H+ antiporter. Here, we used a bioassay-guided isolation approach to identify endogenous small molecules, which affect PM H+-ATPase and Na+/H+ antiporter activities, and found phosphatidylinositol (PI), which inhibits PM H+-ATPase activity in non-stress conditions in Arabidopsis by directly binding to the C-terminus of the PM H+-ATPase AHA2. Under salt stress, the PI4P-to-PI ratio increased, PI4P bound and activated PM Na+/H+ antiporter activity. PI prefers binding to the inactive form of PM H+-ATPase, while PI4P tends to bind the active form of Na+/H+ antiporter. Consistently, pis1 mutants, with reduced levels of PI, displayed increased PM H+-ATPase activity and salt stress tolerance, while pi4kβ1 mutant, with reduced levels of PI4P, displayed reduced PM Na+/H+ antiporter activity and salt stress tolerance. Collectively, we have revealed a dynamic change between PI and PI4P in response to salt stress in Arabidopsis, which is crucial for maintaining ion homeostasis to protect plants from unfavorable environmental conditions.

No abstract available

N6 -methyladenosine (m6 A) is an abundant methylation mark in eukaryotic mRNAs. It is installed and removed by methyltransferases ("writers") and demethylases ("erasers"), respectively. A recent study has demonstrated that alpha-ketoglutarate-dependent dioxygenase homolog 10B (ALKBH10B) is an mRNA m6 A eraser affecting floral transition in Arabidopsis thaliana (A. thaliana). However, the roles of m6 A eraser proteins, including ALKHB10B, in plant adaptation to abiotic stresses are largely unknown. In this study, we aimed to determine the role of ALKBH10B in the response of A. thaliana to abiotic stresses and abscisic acid (ABA). The m6 A level increased in response to salt stress, and m6 A levels in alkbh10b mutants were higher than those in the wild-type under both normal and salt stress conditions. Germination of alkbh10b mutant seeds was markedly delayed under salt stress but not under dehydration, cold, or ABA conditions. Seedling growth and survival rate of alkbh10b mutants were enhanced under salt stress. Notably, salt-tolerant phenotypes of alkbh10b mutants were correlated with decreased levels of several m6 A-modified genes, including ATAF1, BGLU22, and MYB73, which are negative effectors of salt stress tolerance. In response to ABA, both seedling and root growth of alkbh10b mutants were inhibited via upregulating ABA signaling-related genes, including ABI3 and ABI4. Collectively, these findings indicate that ALKBH10B-mediated m6 A demethylation affects the transcript levels of stress-responsive genes, which are important for seed germination, seedling growth, and survival of Arabidopsis in response to salt stress or ABA.

The chloroplast is the main organelle for stress-induced production of reactive oxygen species (ROS). However, how chloroplastic ROS homeostasis is maintained under salt stress is largely unknown. We show that EGY3, a gene encoding a chloroplast-localized protein, is induced by salt and oxidative stresses. The loss of EGY3 function causes stress hypersensitivity while EGY3 overexpression increases the tolerance to both salt and chloroplastic oxidative stresses. EGY3 interacts with chloroplastic Cu/Zn-SOD2 (CSD2) and promotes CSD2 stability under stress conditions. In egy3-1 mutant plants, the stress-induced CSD2 degradation limits H2O2 production in chloroplasts and impairs H2O2-mediated retrograde signaling, as indicated by the decreased expression of retrograde-signal-responsive genes required for stress tolerance. Both exogenous application of H2O2 (or APX inhibitor) and CSD2 overexpression can rescue the salt-stress hypersensitivity of egy3-1 mutants. Our findings reveal that EGY3 enhances the tolerance to salt stress by promoting the CSD2 stability and H2O2-mediated chloroplastic retrograde signaling.

High salinity causes ionic, osmotic, and oxidative stresses to plants, and the antioxidant enzyme Catalase2 (CAT2) plays a vital role in this process, while how CAT2 expression is regulated during plant response to high salinity remains elusive. Here, we report that phytohormone jasmonic acid (JA) impairs plant salt stress tolerance by repressing CAT2 expression in an MYC2-dependent manner. Exogenous JA application decreased plant salt stress tolerance while the jar1 mutant with reduced bioactive JA-Ile accumulation showed enhanced salt stress tolerance. JA enhanced salt-induced hydrogen peroxide (H2O2) accumulation, while treatment with H2O2-scavenger glutathione compromised such effects of JA on plant H2O2 accumulation and salt stress tolerance. In addition, JA repressed CAT2 expression in salt-stressed wild-type plant but not in myc2, a mutant of the master transcriptional factor MYC2 in JA signaling, therefore, the myc2 mutant exhibited increased salt stress tolerance. Further study showed that mutation of CAT2 largely reverted lower reactive oxygen species (ROS) accumulation, higher CAT activity, and enhanced salt stress tolerance of the myc2 mutant in myc2 cat2-1 double mutant, revealing that CAT2 functions downstream JA-MYC2 module in plant response to high salinity. Together, our study reveals that JA impairs Arabidopsis seedling salt stress tolerance through MYC2-mediated repression of CAT2 expression.

Maintenance of root elongation is beneficial for the growth and survival of plants under salt stress, but currently the cellular components involved in the regulation of root growth under high salinity are not fully understood. In this study, we identified an Arabidopsis mutant, rres1, which exhibited reduced root elongation under treatment of a variety of salts, including NaCl, NaNO3, KCl, and KNO3. RRES1 encodes a novel mitochondrial protein and its molecular function is still unknown. Under salt stress, the root meristem length was shorter in the rres1 mutant compared to the wild type, which was correlated with a reduced auxin accumulation in the mutant. Reactive oxygen species (ROS), as important signals that regulate root elongation, were accumulated to higher levels in the rres1 mutant than the wild type after salt treatment. Measurement of monosaccharides in the cell wall showed that arabinose and xylose contents were decreased in the rres1 mutant under salt stress, and application of boric acid, which is required for the crosslinking of pectic polysaccharide rhamnogalacturonan-II (RG-II), largely rescued the root growth arrest of the rres1 mutant, suggesting that RRES1 participates in the maintenance of cell wall integrity under salt stress. GUS staining assay indicated that the RRES1 gene was expressed in leaves and weakly in root tip under normal conditions, but its expression was dramatically increased in leaves and roots after salt treatment. Together, our study reveals a novel mitochondrial protein that regulates root elongation under salt stress via the modulation of cell wall integrity, auxin accumulation, and ROS homeostasis.

Plant stress responses involve dynamic growth regulation. Growth is restricted in harsh environmental conditions and is rapidly restored when conditions improve. Here, we identified BIN2, a glycogen synthase kinase 3 (GSK3)-like kinase, as a molecular switch in the transition to robust growth after salt stress in Arabidopsis thaliana. In the rapid recovery phase after salt stress, the calcium sensors SOS3 and SCaBP8 perceive a calcium signal and promote BIN2 localization to the plasma membrane to repress the salt stress response, and BIN2 inhibits SOS2 activity and enhances growth by releasing BZR1/BES1 transcriptional activity. The expression of stress- and brassinosteroid-responsive genes is coordinately regulated during this process. bin2-3bil1 and bin2-3bil2 mutants defective in BIN2 and its homologs BIL1 and BIL2, respectively, are hyposensitive to salt stress. Our study suggests that salt signaling modulates the subcellular localization and interactions of BIN2. By phosphorylating different substrates, BIN2 regulates the salt stress response and growth recovery.

Glucose-6-phosphate dehydrogenase (G6PD or G6PDH) plays an important role in response to salt stress in plants. However, much less is known about G6PD proteins in soybean (Glycine max L.). Here, we found that a soybean cytosolic G6PD gene, GmG6PD7, was induced by NaCl. We generated Arabidopsis transgenic lines overexpressing GmG6PD7. The seed germination rate and primary root length of Arabidopsis thaliana over-expressing GmG6PD7 under NaCl treatment were enhanced. Salt stress induced an obvious increase of the total and cytosolic G6PD activity and the marked decrease of ROS levels in the transgenic plants. At the same time, over-expressing GmG6PD7 in Arabidopsis affected the glutathione and NADPH level and activated ROS scavengers, suggesting that GmG6PD7 contributes to increase salinity tolerance by decreasing ROS accumulation. What's more, we found GmG6PD7 overexpression led to the up-regulation of abscisic acid (ABA) degradation gene and the down-regulation of ABA synthesis and ABA-responsive genes, which finally reduced ABA content to improve seed germination rate under salinity stress. It was noteworthy that GmG6PD7 can rescue the seed and root phenotype of Arabidopsis cytosolic G6PD mutant (Atg6pd5 and Atg6pd6) under salt stress, suggesting cytosolic G6PD may have a conserved function in soybean and Arabidopsis.

Lateral roots (LRs) are the main component of the root system architecture (RSA) in Arabidopsis. The plasticity of LR development has an important role in improving plant survival in response to the external environment. Previous studies have revealed a number of genetic pathways that control plant growth in response to environmental stimuli. Here, we find that the Xyloglucan Endotransglucosylase 19 (XTH19) and XTH23 genes are involved in LR development under salt stress. LR density was decreased in the xth23 single mutant, which was also more sensitive to salt than the wild type, and xth19xth23 double mutant exhibit additive down-regulated LR initiation and salt sensitivity compared with the single mutant. On the contrary, constitutive overexpression of XTH19 or XTH23 caused increased LR densities. Furthermore, XTH19 and XTH23 were induced by salt via the key BR signaling pathway transcription factor BES1. In addition, we found that 35S::BES1 increased salt tolerance and the phenotype of xth19xth23 & 35S::BES1 was partially complementary to the wild-type level. In vivo and in vitro assays demonstrated that BES1 acts directly upstream of XTH19 and XTH23 to control their expression. Overall, our results revealed that XTH19 and XTH23 are involved in LR development via the BES1-dependent pathway, and contribute to LR adaptation to salt.

Rapid response to environmental changes and abiotic stress to coordinate developmental programs is critical for plants. To accomplish this, plants use the ubiquitin proteasome pathway as a flexible and efficient mechanism to control protein stability and to direct cellular reactions. Here, we show that all three members of the R2R3 S23 MYB transcription factor subfamily, MYB1, MYB25, and MYB109, are degraded by the 26S proteasome, likely facilitated by a CUL3-based E3 ligase that uses MATH-BTB/POZ proteins as substrate adaptors. A detailed description of MYB1, MYB25, and MYB109 expression shows their nuclear localization and specific tissue specific expression patterns. It further demonstrates that elevated expression of MYB25 reduces sensitivities toward abscisic acid, osmotic and salt stress in Arabidopsis, while downregulation of all S23 members results in hypersensitivities. Transcriptional profiling in root and shoot of seedlings overexpressing MYB25 shows that the transcription factor widely affects cellular stress pathways related to biotic and abiotic stress control. Overall, the work extends our knowledge on proteins targeted by CUL3-based E3 ligases that use MATH-BTB/POZ proteins as substrate adaptors and provides first information on all members of the MYB S23 subfamily.

Anthocyanins are a subgroup of plant flavonoids with antioxidant activities and are often induced by various biotic and abiotic stresses in plants, likely to efficiently scavenge free radicals and reactive oxygen species. However, the regulatory mechanisms of salt-stress-induced anthocyanin biosynthesis remain unclear. Using molecular and genetic techniques we demonstrated key roles of ECAP in differential salt-responsive anthocyanin biosynthesis pathways in Arabidopsis thaliana. ECAP, JAZ6/8, and TPR2 are known to form a transcriptional repressor complex, negatively regulate jasmonate (JA)-responsive anthocyanin accumulation. In this study, we demonstrated that under moderate salt stress, the accumulation of anthocyanins is partially dependent on JA signaling, which degrades JAZ proteins but not ECAP. More interestingly, we found that high salinity rather than moderate salinity induced the degradation of ECAP through the 26S proteasome pathway, and this process was independent of JA signaling. Further analysis revealed that ECAP interacts with MYB75 (a transcription factor activating anthocyanin biosynthetic genes) and represses its transcriptional activity in the absence of high salinity. Our results indicated that plants adopt different strategies for fine-tuning anthocyanin accumulation under different levels of salt stress, and further elucidated the complex regulation of anthocyanin biosynthesis during plant development and responses to environmental stresses.

The dehydration-responsive element-binding (DREB) transcription factors play important roles in regulation of plant responses to abiotic stresses, however, few DREBs have been isolated from a desiccation tolerance moss, and the role of DREBs in the DT mechanism is still unknown. We have functionally characterized a unique DREB transcription factor BaDBL1 from the DT moss Bryum argenteum. Expression pattern analysis revealed that BaDBL1 was induced by dehydration-rehydration, salt, cold, and abscisic acid treatments. BaDBL1 was localized in the nucleus and had a transactivation region in its C-terminal region. Overexpression of BaDBL1 in Arabidopsis resulted in significantly increased osmotic and salt stress tolerance, as illustrated by higher fresh weight and antioxidase activities (SOD, POD and CAT) compared with WT under osmotic and salt stresses. Moreover, the transcription of stress-responsive genes, such as AtRD29A and AtCOR15A, AtLEA in BaDBL1-overexpressing lines were significantly up-regulated under osmotic and salt stresses compared with WT. Transcriptomic analysis revealed that BaDBL1-overexpression affected the lignin biosynthesis pathway by improving lignin content and regulating lignin-biosynthesis-related genes under osmotic stress. The results suggest that BaDBL1 may regulate plant tolerance to stress by enhancing anti-oxidase activities, regulating expression of stress-related genes and effecting the lignin biosynthesis, making BaDBL1 a candidate gene for stress tolerance improvement in crops.

ABSTRACT Plant clathrin and its adaptor protein complexes—adaptor protein complex-1 (AP-1) at the trans-Golgi network/early endosome (TGN/EE) and the adaptor protein complex-2 (AP-2) at the plasma membrane (PM)—function in clathrin-mediated trafficking (CMT). This study reports the role of CMT in regulating copper (Cu) tolerance in plants. We found that high concentrations of exogenous Cu treatment increase the abundance of clathrin and adaptor protein complexes at the TGN/EE and/or the PM. We further found that a CMT-deficient mutant ap2μ2, clc2 clc3 exhibits hypersensitivity to Cu stress, similar to a mutant lacking the Cu transporter HEAVY METAL ATPase 5 (HMA5). As previously reported, HMA5 relocates from the endoplasmic reticulum (ER) to the PM on the soil side, where it excretes excess Cu from the root cell, which is crucial for Cu tolerance. Our protein interaction assays showed that the AP-1 and AP-2 σ subunits depend on the YXXΦ sorting motif of HMA5 for recognition. Defective AP-1 hinders HMA5 translocation to the PM after its transfer from the ER to the TGN/EE following Cu stress, while impaired AP-2 function inhibits HMA5 endocytosis at the PM. These results demonstrate that CMT mediates the endocytic recycling of HMA5 between the TGN/EE and the PM, thereby regulating Cu efflux from root cells. Our findings highlight a function of CMT in maintaining Cu homeostasis.

The Ni hyperaccumulator Odontarrhena chalcidica (formerly Alyssum murale), exhibits a significant capacity to accumulate Zn in the roots. However, the molecular mechanisms underlying the variation in Ni and Zn accumulation are poorly understood. Here, we isolated a homolog of heavy metal ATPase 3 from O. chalcidica (OcHMA3) and characterized its functions using heterologous systems. Phylogenetic analysis revealed that OcHMA3 protein shares 87.6 % identity with AtHMA3, with similar metal binding sites to other HMA3 proteins. Heterologous expression of OcHMA3 in yeast increased sensitivity to Cd, Ni and Zn, suggesting it functions as a broad-specificity transporter. Further investigation showed OcHMA3 is constitutively expressed in the roots and localized to the tonoplast. Overexpression of OcHMA3 in A. thaliana shoots increased its roots Zn concentrations by 41.9 % - 74.1 %. However, overexpression of OcHMA3 in roots enhanced its tolerance to Cd and increased roots Cd concentrations by 50.9 % - 90.6 %. Our findings indicated OcHMA3 is responsible for Zn sequestration in root vacuoles, likely leading to Zn retention in roots and subsequent Ni hyperaccumulation in shoots. This study elucidates the molecular mechanism of Ni and Zn accumulation in O. chalcidica, and identifies OcHMA3 as a potential gene for developing Zn-rich plants and for phytoextraction in Cd-contaminated soils.

Cadmium (Cd), which is a nonessential heavy metal element for organisms, can have a severe impact on the growth and development of organisms that absorb excessive Cd. Studies have shown that Brassica carinata, a semiwild oil crop, has strong tolerance to various abiotic stresses, and RNA-seq has revealed that the B. carinata superoxide dismutase gene (BcaSOD1) likely responds to Cd stress. To elucidate the BcaSOD1 function involved in tolerance of Cd stress, we cloned the coding sequences of BcaSOD1 from a purple B. carinata accession and successfully transferred it into Arabidopsis thaliana. The subcellular localization results demonstrated that BcaSOD1 was primarily located in the plasma membrane, mitochondria and nucleus. Overexpression of BcaSOD1 in transgenic Arabidopsis (OE) effectively decreased the toxicity caused by Cd stress. Compared to the WT (wild type lines), the OE lines exhibited significantly increased activities of antioxidant enzymes (APX, CAT, POD, and SOD) after exposure to 2.5 mM CdCl2. The Cd content of underground (root) in the OE line was dominantly higher than that in the WT; however, the Cd content of aboveground (shoot) was comparable between the OE and WT types. Moreover, the qRT‒PCR results showed that several heavy metal detoxification-related genes (AtIREG2, AtMTP3, AtHMA3, and AtNAS4) were significantly upregulated in the roots of OE lines under Cd treatment, suggesting that these genes are likely involved in Cd absorption in the roots of OE lines. In addition, both comparable transcriptome and qRT-PCR analyses revealed that exogenous BcaSOD1 noticeably facilitates detoxification by stimulating the expression of two arginine (Arg) biosynthesis genes (AtGDH1 and AtGDH2) while inhibiting the expression of AtARGAH1, a negative regulator in biosynthesis of Arg. The Arg content was subsequently confirmed to be significantly enhanced in OE lines under Cd treatment, indicating that BcaSOD1 likely strengthened Cd tolerance by regulating the expression of Arg-related genes. This study demonstrates that BcaSOD1 can enhance Cd tolerance and reveals the molecular mechanism of this gene, providing valuable insights into the molecular mechanism of Cd tolerance in plants.

No abstract available

Abstract Transgenic alfalfa (Medicago sativa L.) plants overexpressing the Arabidopsis ATP sulfurylase gene were generated using Agrobacterium-mediated genetic transformation to enhance their heavy metal accumulation efficiency. The ATP sulfurylase gene was cloned from Arabidopsis, following exposure to vanadium (V) and lead (Pb), and transferred into an Agrobacterium tumefaciens binary vector. This was co-cultivated with leaf explants of the alfalfa genotype Regen SY. Co-cultivated leaf explants were cultured on callus and somatic embryo induction medium, followed by regeneration medium for regenerating complete transgenic plants. The transgenic nature of the plants was confirmed using PCR and southern hybridization. The expression of Arabidopsis ATP sulfurylase gene in the transgenic plants was evaluated through RT-PCR. The selected transgenic lines showed increased tolerance to a mixture of five heavy metals and also demonstrated enhanced metal uptake ability under controlled conditions. The transgenic lines were fertile and did not exhibit any apparent morphological abnormality. The results of this study indicated an effective approach to improve the heavy metal accumulation ability of alfalfa plants which can then be used for the remediation of contaminated soil in arid regions.

The Endoplasmic Reticulum Quality Control (ERQC) machinery is highly conserved among eukaryotes and assists the newly synthetized proteins in the folding process. Previous works have reported the involvement of ERQC in plant immunity and biotic stress response. However, the interaction between ERQC pathway and heavy metals exposure has been poorly investigated in plants. In the present study, we showed that the Arabidopsis thaliana rsw3 mutant, characterised by a reduced activity of the ER Glucosidase II enzyme, exhibits an increased tolerance to cadmium (Cd) stress. Under standard conditions, rsw3 seedlings exhibit shorter primary roots compared to Wild-type (Wt) plantlets, because of a constitutive ER stress and a consequent upregulation of both ERQC and Unfolded Protein Response (UPR) stress markers in root or shoot tissues. Interestingly, differently from Wt seedlings, these markers remain unchanged in rsw3 under Cd stress. Biochemical data here provided linked the enhanced Cd tolerance of rsw3 to the brassinosteroid receptor 1, BRI1, as the partial impairment of GII activity positively affects the accumulation of the active form of BRI1 receptor on the plasma membrane under Cd stress.

Cysteine (Cys) is an essential amino acid component of the major heavy metal chelators, such as glutathione (GSH), metallothioneins (MTs), and phytochelatins (PCs), which are involved in the pathways of mercury (Hg) tolerance in plants. However, the mechanism through which Cys facilitates Hg tolerance in plants remains largely unclear. In this study, we investigated the effects of exogenous Cys on Hg uptake in the seedlings, roots, and shoots of Arabidopsis throughout 6 and 36 h of Hg exposure and on the regulation of Hg detoxification by heavy metal chelators and antioxidative enzymes. The results showed that exogenous Cys significantly improved Hg tolerance during the germination and seedling growth stages in Arabidopsis. Exogenous Cys significantly promoted Hg uptake in Arabidopsis roots by upregulating the expression of the Cys transporter gene AtLHT1, resulting in increased Hg accumulation in the roots and seedlings. In Arabidopsis seedlings, exogenous Cys further increased the Hg-induced glutathione synthase (GS1 and GS2) transcript levels, and the Hg and Hg + Cys treatments greatly upregulated MT3 expression after 36 h exposure. In the roots, MT3 was also significantly upregulated by treatment of 36 h of Hg or Hg + Cys. Notably, in the shoots, MT2a expression was rapidly induced (10-fold) in Hg presence and further markedly increased (20-fold) by exogenous Cys. Moreover, in the seedlings, exogenous Cys upregulated the transcripts of all superoxide dismutase (CuSOD1, CuSOD2, MnSOD1, FeSOD1, FeSOD2, and FeSOD3) within 6 h and subsequently increased the Hg-induced GR1 and GR2 transcript levels at 36 h, all of which could eliminate the promotion of reactive oxygen species production and cell damage caused by Hg. Additionally, exogenous Cys upregulated all the antioxidative genes rapidly in the roots and subsequently increased the expression of CuSOD1, CuSOD2, and MnSOD1 in the shoots. These results indicate that exogenous Cys regulates the transcript levels of heavy metal chelators and antioxidative enzymes differently in a time- and organ-specific manner under Hg stress. Taken together, our study elucidates the positive functional roles of exogenous Cys in the Hg uptake and tolerance mechanisms of Arabidopsis.

Anthropogenic activities cause the leaching of heavy metals into groundwater and their accumulation in soil. Excess levels of heavy metals cause toxicity in plants, inducing the production of reactive oxygen species (ROS) and possible death caused by the resulting oxidative stress. Heavy metal stresses repress auxin biosynthesis and transport, inhibiting plant growth. Here, we investigated whether nickel (Ni) heavy metal toxicity is reduced by exogenous auxin application and whether Ni stress tolerance in Arabidopsis thaliana is mediated by the bifunctional enzyme YUCCA6 (YUC6), which functions as an auxin biosynthetic enzyme and a thiol-reductase (TR). We found that an application of up to 1 µM exogenous indole-3-acetic acid (IAA) reduces Ni stress toxicity. yuc6-1D, a dominant mutant of YUC6 with high auxin levels, was more tolerant of Ni stress than wild-type (WT) plants, despite absorbing significantly more Ni. Treatments of WT plants with YUCASIN, a specific inhibitor of YUC-mediated auxin biosynthesis, increased Ni toxicity; however yuc6-1D was not affected by YUCASIN and remained tolerant of Ni stress. This suggests that rather than the elevated IAA levels in yuc6-1D, the TR activity of YUC6 might be critical for Ni stress tolerance. The loss of TR activity in YUC6 caused by the point-mutation of Cys85 abolished the YUC6-mediated Ni stress tolerance. We also found that the Ni stress–induced ROS accumulation was inhibited in yuc6-1D plants, which consequently also showed reduced oxidative damage. An enzymatic assay and transcriptional analysis revealed that the peroxidase activity and transcription of PEROXIREDOXIN Q were enhanced by Ni stress to a greater level in yuc6-1D than in the WT. These findings imply that despite the need to maintain endogenous IAA levels for basal Ni stress tolerance, the TR activity of YUC6, not the elevated IAA levels, plays the predominant role inNi stress tolerance by lowering Ni-induced oxidative stress.

Abiotic stresses such as high temperature, high humidity, and heavy metals are important factors that affect seed development and quality, and restrict yield in soybean. The ATX1-type copper chaperones are an important type of proteins that are used for maintaining intracellular copper ion homeostasis. In our previous study, a copper chaperone protein GmATX1 was identified in developing seeds of soybean under high temperature and humidity (HTH) stresses. In this study, the GmATX1 gene was isolated, and multiple alignment analysis showed that its encoding protein shared high sequence identities with other plant orthologues of copper chaperone proteins containing the HMA domain, and a conserved metal ion-binding site, CXXC. A subcellular localization assay indicated that GmATX1 was localized in the cell membrane and nucleus. An expression analysis indicated that GmATX1 was involved in seed development, and in response to HTH and heavy metal stresses in soybean. GmATX1-silent soybean seedlings were found to be more severely damaged than the control under HTH stress. Moreover, the silencing of GmATX1 reduced antioxidase activity and reactive oxygen species (ROS) scavenging ability in the seedling leaves. The overexpression of GmATX1 in Arabidopsis improved seed vigor and seedling tolerance, and enhanced antioxidase activity and ROS scavenging ability under HTH and heavy metal stresses. Our results indicated that GmATX1 could promote seed vigor and seedling tolerance to HTH and heavy metal stresses in transgenic Arabidopsis, and this promotion could be achieved by enhancing the antioxidase activity and ROS scavenging ability.

ABSTRACT Plant growth-promoting rhizobacteria (PGPRs) have been utilized to immobilize heavy metals, limiting their translocation in metal contaminated settings. However, studies on the mechanisms and interactions that elucidate how PGPRs mediate Nickel (Ni) tolerance in plants are rare. Thus, in this study we investigated how two pre-characterized heavy metal tolerant isolates of Morganella morganii (ABT9 and ABT3) improve Ni stress tolerance in Arabidopsis while enhancing its growth and yield. Arabidopsis seedlings were grown for five weeks in control/Ni contaminated (control, 1.5 mM and 2.5 mM) potted soil, in the presence or absence of PGPRs. Plant growth characteristics, quantum yield, and antioxidative enzymatic activities were analyzed to assess the influence of PGPRs on plant physiology. Oxidative stress tolerance was quantified by measuring MDA accumulation in Arabidopsis plants. As expected, Ni stress substantially reduced plant growth (shoot and root fresh weight by 53.25% and 58.77%, dry weight by 49.80% and 57.41% and length by 47.16% and 64.63% over control), chlorophyll content and quantum yield (by 40.21% and 54.37% over control). It also increased MDA content by 84.28% at higher (2.5 mM) Ni concentrations. In contrast, inoculation with M. morganii led to significant improvements in leaf chlorophyll, quantum yield, and Arabidopsis biomass production. The mitigation of adverse effects of Ni stress on biomass observed in M. morganii-inoculated plants was attributed to the enhancement of antioxidative enzyme activities compared to Ni-treated plants. This upregulation of the antioxidative defense mechanism mitigated Ni-induced oxidative stress, leading to improved performance of the photosynthetic machinery, which, in turn, enhanced chlorophyll content and quantum yield. Understanding the underlying mechanisms of these tolerance-inducing processes will help to complete the picture of PGPRs-mediated defense signaling. Thus, it suggests that M. morganii PGPRs candidate can potentially be utilized for plant growth promotion by reducing oxidative stress via upregulating antioxidant defense systems in Ni-contaminated soils and reducing Ni metal uptake.

Glutathione S-transferases (GSTs) are well-known enzymes due to their role in detoxification of xenobiotic compounds. However, their biochemical action is still not so clear in imparting tolerance to several abiotic stresses in crop plants. In our previous study, we observed that rice tau class OsGSTU30 plays a significant role in the detoxification of Cr(vi). Interestingly, q-RT PCR analysis also revealed higher expression of OsGSTU30 under drought conditions. In this study, we characterize OsGSTU30 in response to drought as well as heavy metal [Cr(vi)] stresses through overexpression in Arabidopsis thaliana. Biochemical and physiological analyses revealed that OsGSTU30 overexpression lines have improved tolerance against both stresses as compared to wild-type plants. Kinetic analysis and molecular docking confirmed that OsGSTU30 enzyme possesses both GST as well as glutathione peroxidase (GPx) like activity. Differentially expressed stress-responsive genes were also identified by transcriptome analysis, involved in different biological pathways during abiotic stresses. These results suggest the signaling functions of OsGSTU30 apart from its catalytic activity during abiotic stress responses and can be further exploited for improving the stress tolerance in crops.

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

No abstract available

Heavy metal stress is a critical challenge to agricultural productivity, necessitating deeper insights into the molecular mechanisms of metal transport in plants. In this study, we conducted a comprehensive genome-wide characterization of the Natural Resistance-Associated Macrophage Protein (NRAMP) gene family in Arabidopsis thaliana and identified six AtNRAMP genes. Phylogenetic and synteny analyses revealed their distribution into two distinct clades and evolutionary conservation with legumes such as Glycine max and Arachis hypogaea, indicating functional divergence and gene duplication events maintained under purifying selection. Conserved protein motifs and domains, particularly the NRAMP transmembrane domain, highlighted their conserved role in divalent metal ion transport, while cis-regulatory element analysis demonstrated enrichment of stress- and hormone-responsive elements, pointing to tight transcriptional regulation under environmental challenges. Structural modeling further supported the functional conservation of AtNRAMP proteins. Expression profiling showed clear tissue-specific expression under normal conditions and strong, differential regulation in response to cadmium and other heavy metals, as well as to the phytohormone abscisic acid (ABA). Collectively, these results provide foundational insights into the evolutionary relationships, regulatory mechanisms, and stress-responsive expression of the AtNRAMP gene family, offering a framework for future functional studies and potential applications in developing crops with enhanced heavy metal tolerance and improved growth under stress conditions.

The pollution of heavy metals is extremely serious in China, including zinc (Zn), copper (Cu), lead (Pb), and cadmium (Cd). Heavy-metal-transporting ATPase (HMA) belongs to a subfamily of the P-ATPase family, which absorbs and transports Zn, Cu, Pb, and Cd in plants. Here, we describe a ZmHMA-encoding HMA family protein that positively regulates Cd and Zn tolerance. The real-time fluorescence quantification (RT-PCR) results revealed that ZmHMA3 had a high expression in B73, and the expression of ZmHMA3 was sensitive to Cd in yeast cells, which was related to Cd accumulation in yeast. Additionally, the Arabidopsis thaliana homologous mutants of AtHMA2 showed Cd sensitivity compared with WT. The overexpressing ZmHMA3 plants showed higher tolerance under Cd and Zn stresses than the wild type. The overexpression of ZmHMA3 led to higher Cd and Zn accumulation in tissues based on the subcellular distribution analysis. We propose that ZmHMA3 improves maize tolerance to Cd and Zn stresses by absorbing and transporting Cd and Zn ions. This study elucidates the gene function of the ZmHMA3 response to Cd and Zn stress and provides a reference for improving the characteristics of heavy metals enrichment in existing maize varieties and the plant remediation technology of heavy-metal-contaminated soil.

No abstract available

Chickpea (Cicer arietinum L.), a widely cultivated legume, is valued for its high protein and mineral content. However, its production has been inconsistent in recent years, largely due to biotic and abiotic stresses and limited genetic diversity. Micronutrient deficiencies, particularly of iron and zinc, remain a global challenge and are often referred to as “hidden hunger”. The NRAMP (natural resistance- associated macrophage protein) gene family plays a crucial role in the uptake and transport of heavy metals such as cadmium (Cd), zinc (Zn), copper (Cu), lead (Pb), iron (Fe), and manganese (Mn), and contributes significantly to plant responses under heavy metal stress conditions. In this study, a genome-wide analysis was conducted to identify and characterize NRAMP genes in the chickpea genome using bioinformatics approaches. Multiple sequence alignment was performed using the ClustalW method in MEGA 7 to examine conserved residues across NRAMP proteins from Cicer arietinum, Nicotiana attenuata, Arabidopsis thaliana, Oryza sativa, Solanum lycopersicum, and Solanum tuberosum. A total of nine NRAMP genes were identified in chickpea. The phylogenetic analysis grouped these genes into five distinct clades. Additionally, physicochemical profiling revealed that Ca-NRAMP6 has the longest protein sequence, while Ca-NRAMP4 has the highest number of exons and introns. Protein interaction analysis indicated that only CaNRAMP 6 has a strong interaction with other proteins.

No abstract available

Glutathione (GSH) plays a crucial role in cellular redox regulation and enhancing plant survival under heavy metal toxicity. In plants, GSH homeostasis is managed through the γ-glutamyl cycle, though the genes involved in GSH degradation and recycling are not fully understood. Here, we have characterized the Arabidopsis oxoprolinase 1 (AtOXP1) gene for its role in providing tolerance to arsenite (AsIII) and mercury (Hg) stress. The oxp1 T-DNA mutants showed severe sensitivity to Hg, with significantly lower shoot (~40%) and root (~86%) biomass compared with WT. The mutants accumulated ~45% higher arsenic and Hg accumulation in shoots. The oxp1 mutants also had elevated GSH (2.3-4.0 folds) and oxoproline (5-OP) levels (96%-265%) compared with Col-0 under control and stress conditions. The mutants showed significantly (15%-78%) lower glutamate levels than Col-0 under metal stress. Conversely, relative to WT, AtOXP1 overexpression (OE) lines had significantly higher biomass (18%-43% more) and reduced As (40%) and Hg (19%-35%) accumulation in the shoots. Compared with Col-6, the AtOXP1 OE lines had significantly (44%-57%) lower buildup of GSH levels under both AsIII and Hg treatment. Isotopic analysis revealed that the oxp1 mutants had 29% higher 15N-Glu/Total Glu compared to WT, while OE lines were similar to WT. Additionally, OE lines had significantly higher shoot (15%-22%) and root (~77%) biomass under low nitrogen conditions. The results demonstrate that OXP1 is involved in Glu recycling via GSH degradation and holds the potential to improve plant productivity and reduce toxic metal(loid)s accumulation for food safety.

In China, soil contamination by heavy metals is a widespread issue, with substantial increases in lead(Pb), cadmium(Cd), copper(Cu), and zinc(Zn) levels observed across various regions. Particularly, the concentrations of Pb and Cd significantly exceed their natural background levels. P-ATPases, a group of proteins, utilize energy from ATP hydrolysis to support the transmembrane movement of metal ions. This group encompasses several Heavy Metal Associated Transporter (HMA) ATPases. Studies on hyperaccumulators have shown the critical role of HMAs in the movement and reduction in Zn and Cd toxicity in plant systems. This research identifies a protein encoded by the SpHMA gene from Sedum plumbizincicola, a species noted for aiding Zn/Cd hyperaccumulators, which enhances tolerance to Cd and Zn. We detail a protein encoded by SpH/A within the HMA family that enhances Cd tolerance. Real-time fluorescence quantification (RT-PCR) indicates that SpHMA3 expression in Arabidopsis thaliana and Zea mays KN5585 correlates with high Cd tolerance, linked to Cd accumulation in Zea mays. In addition, homozygous Arabidopsis thaliana AtHMA3 mutants exhibited increased Cd sensitivity compared to the wild type (WT). Notably, plants of Arabidopsis thaliana and maize overexpressing SpHMA3 showed enhanced Cd stress tolerance compared to WT. Enhanced Cd accumulation in tissues was observed when SpHMA3 was overexpressed, as revealed by subcellular distribution analysis. We propose that SpHMA3 augments maize tolerance to Cd and Zn stresses through enhanced cellular uptake and translocation of Cd ions. This investigation clarifies the gene function of SpHMA3 in Cd and Zn stress response, offering insights for enhancing heavy metal absorption traits in maize varieties and phytoremediation methods for soils contaminated with heavy metals.

Cadmium (Cd), as a widely investigated heavy metal, demonstrates severe toxicity to plant growth and development. However, the roles of energy metabolism, and protein degradation caused by autophagy in plant Cd tolerance remain poorly characterized. This study reveals that Cd-exposure simultaneously affects both the glycolytic and autophagic in Arabidopsis thaliana. Cd-exposure enhances autophagy by upregulating the mRNA and protein level of AtATG7 and finally promotes the autophagy-induced energy burst. Another hand, Cd-exposure induces the accumulation of AtGAPDH and AtENO2, further boosting energy production during glycolysis. Through treatments with the autophagic modulators rapamycin and 3-methyladenine, and the autophagy-deficient mutant Atatg7, it was found that Cd-exposure establishes a link between autophagy and glycolysis through the autophagic degradation of AtHK2. Furthermore, using non-invasive micro-test technology, both the autophagic degradation of AtHK2 and the enhancement of glycolysis significantly promote Cd²⁺ efflux in Arabidopsis. Finally, the changes of ATP level demonstrate that the synergistic mediation of autophagy and glycolysis provides sufficient energy for the efflux of Cd2 + under Cd-exposure. These findings elucidate the molecular mechanisms underlying Cd tolerance in Arabidopsis through synergistic regulation of autophagy and glycolysis pathways, and offer crucial theoretical foundations and novel perspectives for developing plant-based remediation strategies for Cd-contaminated soils.

Unlike terrestrial angiosperm plants, the freshwater aquatic angiosperm duckweed (Spirodela polyrhiza) grows directly in water and has distinct responses to heavy-metal stress. Plantlets accumulate metabolites, including lipids and carbohydrates, under heavy-metal stress, but how they balance metabolite levels is unclear, and the gene networks that mediate heavy-metal stress responses remain unknown. Here, we show that heavy-metal stress induced by flue gas desulfurization (FGD) wastewater reduces chlorophyll contents, inhibits growth, reduces membrane lipid biosynthesis, and stimulates membrane lipid degradation in S. polyrhiza, leading to triacylglycerol and carbohydrate accumulation. In FGD wastewater-treated plantlets, the degraded products of monogalactosyldiacylglycerol, primarily polyunsaturated fatty acids (18:3), were incorporated into triacylglycerols. Genes involved in early fatty acid biosynthesis, β-oxidation, and lipid degradation were upregulated while genes involved in cuticular wax biosynthesis were downregulated by treatment. The transcription factor gene WRINKLED3 (SpWRI3) was upregulated in FGD wastewater-treated plantlets, and its ectopic expression increased tolerance to FGD wastewater in transgenic Arabidopsis (Arabidopsis thaliana). Transgenic Arabidopsis plants showed enhanced glutathione and lower malondialdehyde contents under stress, suggesting that SpWRI3 functions in S. polyrhiza tolerance of FGD wastewater-induced heavy-metal stress. These results provide a basis for improving heavy metal-stress tolerance in plants for industrial applications.

Background: Abiotic stresses, such as drought, salinity, temperature fluctuations, waterlogging, and heavy metal contamination, have a detrimental impact on plants, leading to reduced global agricultural productivity. The accumulation of cadmium (Cd) and arsenic (As) in agricultural soil, resulting from both natural and anthropogenic activities, poses significant threats to crop production and food safety. Dehydrins, also known as Group II Late Embryogenesis Abundant (LEA) proteins, are intrinsically disordered proteins that play crucial roles in protecting cellular structures during abiotic stress conditions. These proteins are considered promising candidates for enhancing plant tolerance to environmental stresses through their membrane-stabilizing and protective functions. Methods: This study evaluated the tolerance of Arabidopsis transgenic lines expressing a bacterial dehydrin gene (BG757) to Cd and As stresses using various physiological and biochemical parameters. Results: Compared with the wild-type (WT) control, the transgenic line (35S::BG757-1/Col-0) displayed significant increases in root and shoot growth upon exposure to Cd and As. Furthermore, transgenic plants exposed to heavy metal stress exhibited higher concentrations of chlorophyll, total protein, free proline, total flavonoid, and total phenolic content compared to WT plants. Likewise, transgenic plants showed higher 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and retained higher relative water content under stress conditions. Conclusions: Taken together, these findings suggest that bacterial dehydrins confer enhanced tolerance to heavy metal stress in transgenic Arabidopsis plants, highlighting their potential application in developing stress-resilient crops for contaminated environments.

Soil zinc (Zn) contamination poses a serious threat to living organisms. This study investigated the function of the glutathione (GSH) synthetase gene PsGSH1 from Potentilla sericea in Zn stress response. Transcriptome analysis revealed that Zn stress markedly activated the GSH metabolic pathway and induced PsGSH1 expression in P. sericea. The promoter region of PsGSH1 contains multiple stress-responsive cis-acting elements. Functional assays demonstrated that PsGSH1 overexpression significantly enhanced Zn tolerance in yeast and Arabidopsis thaliana. Under Zn stress, transgenic A. thaliana exhibited increased antioxidant enzyme activities (superoxide dismutase, peroxidase, catalase), higher levels of GSH and proline, and lower oxidative damage indicators (malondialdehyde, superoxide anion, hydrogen peroxide, and relative electrolyte leakage). Furthermore, expression of heavy metal detoxification-related genes (AtGSH1, AtGSH2, AtPCS1, AtPCS2) was significantly up-regulated. These findings suggest that PsGSH1 mediates Zn tolerance by enhancing antioxidant defenses and chelation capacity, providing a potential candidate gene for developing Zn-tolerant plants.

Arsenate [As(V)], an inorganic form of heavy metal Arsenic (As), severely affects plant growth and overall development. As(V) is structurally analogous to Phosphate (Pi) and its uptake is mediated through the plasma membrane localised Pi transporters in the root. As(V) uptake is enhanced in Pi-deficient conditions and causes severe oxidative damage at the cellular level. The Raffinose Family Oligosaccharides (RFOs), including galactinol, function as osmoprotectants in plants in As(V) stress conditions, yet their precise role remains unclear. This study demonstrates that the rate-limiting enzyme of the RFO biosynthesis pathway, encoded by AtGolS1, a galactinol synthase gene, modulates As(V) stress tolerance in Arabidopsis. Comparative analyses between As(V)-tolerant ecotype Col-0 and sensitive Slavi-1 revealed significantly higher AtGolS1 expression in Col-0 under As(V) and Low Phosphate (Pi)+As(V) stress. Transgenic lines overexpressing AtGolS1 in the Slavi-1 background exhibited enhanced root and shoot growth and reduced reactive oxygen species (ROS) accumulation under As(V) stress, accompanied by elevated galactinol levels. Molecular analyses showed that AtGolS1OX lines downregulated high-affinity Pi transporters (PHT1;1, PHT1;4) and regulators such as PHO1 and PHF1, likely restricting As(V) uptake via Pi transport pathways. Under As(V) stress, AtGolS1OX lines accumulated less Pi and As than Slavi-1. Detoxification genes AtABCC1 and AtABCC2 were more strongly expressed in AtGolS1OX lines than in Slavi-1, suggesting improved vacuolar sequestration of As. A single amino acid substitution in Slavi-1 AtGolS1 did not alter its catalytic domain, implicating transcriptional regulation as the key difference. These results identify AtGolS1 as a critical node linking galactinol metabolism to Pi transporter regulation, thereby mitigating As(V) toxicity in Arabidopsis.

The rare earth element lanthanum (La) as a heavy metal can cause stress damage to plants at high concentrations. Metallothioneins (MTs) are low molecular weight proteins rich in cysteine and play significant roles in detoxification of metal ion stress in plants. In this study, we identified a novel function of CnMT2 from Chrysanthemum naktongense. Two MT homologous genes, namely CnMT2 and CnMT3, were cloned from the wild plant species C. naktongense, which exhibits high resistance to the rare earth element lanthanum (La). qRT-PCR analysis revealed that in C. naktongense, the highest expression level of CnMT2 occurred in roots. Its expression in stems, leaves, or shoot tips was also detected. Low expression levels of CnMT3 occurred in all tissues, with relatively higher expression levels in roots. LaCl3 stress treatment could induce the expression of both CnMT2 and CnMT3, resulting in differential expression patterns between shoots and roots. Transgenic Arabidopsis seeds overexpressing CnMT2 exhibited increased resistance to LaCl3 and H2O2 during germination. Transgenic seedlings also exhibited growth enhancement with prolonged LaCl3 stress, accompanied by significantly elevated enzyme activities of catalase (CAT) and peroxidase (POD). Our data revealed that CnMT2 from C. naktongense was able to enhance plant resistance to lanthanum and antioxidative capacity. Overall, the study unveiled a new function of CnMT2 in C. naktongense, which can be potentially used for engineering genotypes for phytoremediation.

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Cadmium (Cd) represents a hazardous heavy metal, prevalent in agricultural soil due to industrial and agricultural expansion. Its propensity for being absorbed by edible plants, even at minimal concentrations, and subsequently transferred along the food chain poses significant risks to human health. Accordingly, it is imperative to investigate novel genes and mechanisms that govern Cd tolerance and detoxification in plants. Here, we discovered that the transcription factor MYC2 directly binds to the promoters of HMA2 and HMA4 to repress their expression, thereby altering the distribution of Cd in plant tissues and negatively regulating Cd stress tolerance. Additionally, molecular, biochemical, and genetic analyses revealed that MYC2 interacts and cooperates with MYB43 to negatively regulate the expression of HMA2 and HMA4 and Cd stress tolerance. Notably, under Cd stress conditions, MYC2 undergoes degradation, thereby alleviating its inhibitory effect on HMA2 and HMA4 expression and plant tolerance to Cd stress. Thus, our study highlights the dynamic regulatory role of MYC2, in concert with MYB43, in regulating the expression of HMA2 and HMA4 under both normal and Cd stress conditions. These findings present MYC2 as a promising target for directed breeding efforts aimed at mitigating Cd accumulation in edible plant roots.

Cadmium (Cd) is a significant heavy metal contaminant within the environment, carrying a notable level of toxicity that presents a substantial hazard to both plant and human. Carrot (Daucus carota), a significant root vegetable crop globally, have evolved multiple transcriptional regulatory mechanisms to cope with Cd stress, with a crucial involvement of the myeloblastosis (MYB) transcription factor. In this study, the DcMYB62 gene encoding 288 amino acids, localized in the nucleus and demonstrated transcription activation property, was isolated from carrot (cv. 'Kuroda'). There was a positive relationship observed between the levels of DcMYB62 expression and the accumulation patterns of carotenoids in two distinct carrot cultivars. Further investigation revealed that the expression of DcMYB62 improved Cd tolerance of Arabidopsis by increasing seed germination rate, root length, and overall survival rate. The levels of carotenoids in DcMYB62 transgenic Arabidopsis surpassed those in wild type, accompanied by elevated expression levels of 15-cis-phytoene desaturase, zeta-carotene desaturase, and carotenoid isomerase. Meanwhile, the heterologous expression of DcMYB62 promoted the biosynthesis of abscisic acid (ABA) and hydrogen sulfide (H2S), which in turn suppressed the formation of hydrogen peroxide and superoxide anion, while also stimulating stomatal closure. Furthermore, the heterologous expression of DcMYB62 increased the transcription of genes associated with heavy metal resistance in Arabidopsis, notably nicotianamine synthase. Overall, this study contributes to understanding how DcMYB62 promote Cd stress resistance of plants by regulating the biosynthesis pathways of carotenoids, ABA, and H2S, which offers valuable insights into the regulatory mechanism connecting DcMYBs with Cd stress response of carrot.

Cadmium (Cd), as a heavy metal, not only negatively affects the development and yield of plants, but also threatens human health due to its accumulation in plants. Increasing evidences indicate that the JUMONJI-C DOMAIN-CONTAINING PROTEIN (JMJ) gene family plays a key role in regulating plant development and stress. Therefore, in this study, SlJMJ524, a 1254 bp gene encoding the jumonji C domain (417 amino acids), was highly expressed in tomato leaves and flowers. Interestingly, the transgenic plants exhibited sensitivity to Cd during post-germination stage but showed enhanced tolerance to the heavy metal during adult stage. Overexpression of SlJMJ524 increased the expression level of related proteins gene involved in heavy metal uptake while increasing Cd tolerance through the GSH-PC pathway. The higher transcription of genes related to flavonoid synthesis reflected higher accumulations of flavonoids in transgenic plants. Our study demonstrated that the ectopic expression of SlJMJ524 conferred the transgenic plants many traits for improving cadmium stress tolerance at different developmental stages. This study advances our collective understanding of the functional role of JMJs and can be used to improve the cadmium tolerance and breeding of crops and plants.

The toxic heavy metal cadmium (Cd) restricts plant growth. However, how plants fine-tune their growth to modulate Cd resistance has not been determined. Ethylene response factors (ERFs) are key regulators of Cd stress, and Arabidopsis thaliana ERF13 and ERF6 (AtERF13 and AtERF6) negatively regulate growth. We previously demonstrated that AtERF13 is a transcriptional activator that binds a Cd-responsive element. Herein, we report that Arabidopsis plants improve Cd tolerance by repressing AtERF13 and AtERF6. We found that AtERF13 and AtERF6 were strongly downregulated by Cd stress and that AtERF6 weakly bound Cd-responsive elements. Moreover, AtERF13 physically interacted with AtERF6. Importantly, AtERF13 and AtERF6 double knockout mutants, but not single mutants or overexpression lines, grew better, tolerated more Cd and had higher Cd contents than did the wild type. Comparative transcriptome analysis revealed that the double mutants regulate the defense response to cope with Cd toxicity. Accordingly, we propose that, upon Cd stress, Arabidopsis plants repress AtERF13 and AtERF6 to relieve their growth inhibition effects and adjust the transcriptome to adapt to Cd stress, leading to increased Cd tolerance. Our findings thereby provide deep mechanical insights into how dual-function transcription factors fine-tune growth and the transcriptome to promote Cd tolerance in plants.

White clover (Trifolium repens L.) is an important forage and aesthetic plant species, but it is susceptible to drought and heat stress. The phytohormone auxin regulates several aspects of plant development and alleviates the effects of drought stress in plants, including white clover, by involving auxin/indole acetic acid (Aux/IAA) family genes. However, Aux/IAA genes and the underlying mechanism of auxin-mediated drought response remain elusive in white clover. To extend our understanding of the multiple functions of Aux/IAAs, the current study described the characterization of a member of the Aux/IAA family TrIAA27 of white clover. TrIAA27 protein had conserved the Aux/IAA family domain and shared high sequence similarity with the IAA27 gene of a closely related species and Arabidopsis. Expression of TrIAA27 was upregulated in response to heavy metal, drought, salt, NO, Ca2+, H2O2, Spm, ABA, and IAA treatments, while downregulated under cold stress in the roots and leaves of white clover. TrIAA27 protein was localized in the nucleus. Constitutive overexpression of TrIAA27 in Arabidopsis thaliana led to enhanced hypocotyl length, root length, plant height, leaf length and width, and fresh and dry weights under optimal and stress conditions. There was Improved photosynthesis activity, chlorophyll content, survival rate, relative water content, endogenous catalase (CAT), and peroxidase (POD) concentration with a significantly lower electrolyte leakage percentage, malondialdehyde (MDA) content, and hydrogen peroxide (H2O2) concentration in overexpression lines compared to wild-type Arabidopsis under drought and salt stress conditions. Exposure to stress conditions resulted in relatively weaker roots and above-ground plant growth inhibition, enhanced endogenous levels of major antioxidant enzymes, which correlated well with lower lipid peroxidation, lower levels of reactive oxygen species, and reduced cell death in overexpression lines. The data of the current study demonstrated that TrIAA27 is involved in positively regulating plant growth and development and could be considered a potential target gene for further use, including the breeding of white clover for higher biomass with improved root architecture and tolerance to abiotic stress.

Heavy metal exposure is a serious environmental stress in plants. However, plants have evolved several strategies to improve their heavy metal tolerance. Heavy metal-associated proteins (HMPs) participate in heavy metal detoxification. Here, we identified 46 and 55 HMPs in rice and Arabidopsis, respectively, and named them OsHMP 1–46 and AtHMP 1–55 according to their chromosomal locations. The HMPs from both plants were divided into six clades based on the characteristics of their heavy metal-associated domains (HMA). The HMP gene structures and motifs varied greatly among the different classifications. The HMPs had high collinearity and were segmentally duplicated. A cis-element analysis revealed that the HMPs may be regulated by different transcription factors. An expression profile analysis disclosed that only eight OsHMPs were constitutive in rice tissues. Of these, the expression of OsHMP37 was far higher than that of the other seven genes while OsHMP28 was expressed exclusively in the roots. For Arabidopsis, nine AtHMPs presented with very high transcript levels in all organs. Most of the selected OsHMPs were differentially expressed in various tissues under different heavy metal stresses. Only OsHMP09, OsHMP18, and OsHMP22 showed higher expression levels in all tissues under different heavy metal stresses. In contrast, most of the selected AtHMPs had nearly constant expression levels in different tissues under various heavy metal stresses. The AtHMP20, AtHMP23, AtHMP25, AtHMP31, AtHMP35, AtHMP46 expression levels under different heavy metal stresses were higher in the leaves and roots. The foregoing discoveries elucidated HMP evolution in monocotyledonous and dicotyledonous plants and may helpful functionally characterize HMPs in the future.

Plant glycine-rich RNA-binding proteins (GRPs) play crucial roles in the response to environmental stresses. However, the functions of AtGRP7 in plants under heavy metal stress remain unclear. In the present study, in Arabidopsis, the transcript level of AtGRP7 was markedly increased by Ni but was decreased by Pb. AtGRP7-overexpressing plants improved Ni tolerance, whereas the knockout mutant (grp7) was more susceptible than the wild type to Ni. In addition, grp7 showed greatly enhanced Pb tolerance, whereas overexpression lines showed high Pb sensitivity. Ni accumulation was reduced in overexpression lines but increased in grp7, whereas Pb accumulation in grp7 was lower than that in overexpression lines. Ni induced glutathione synthase genes GS1 and GS2 in overexpression lines, whereas Pb increased metallothionein genes MT4a and MT4b and phytochelatin synthase genes PCS1 and PCS2 in grp7. Furthermore, Ni increased CuSOD1 and GR1 in grp7, whereas Pb significantly induced FeSOD1 and FeSOD2 in overexpression lines. The mRNA stability of GS2 and PCS1 was directly regulated by AtGRP7 under Ni and Pb, respectively. Collectively, these results indicate that AtGRP7 plays a crucial role in Ni and Pb tolerance by reducing Ni and Pb accumulation and the direct or indirect post-transcriptional regulation of genes related to heavy metal chelators and antioxidant enzymes.

Cadmium (Cd) is a harmful heavy metal that is highly toxic to plants and animals. Expansins are cell wall proteins inducing cell wall loosening and participate in all plant growth and development processes which are associated with cell wall modifications. We investigated lettuce's expansin gene LsEXPA6 and found that LsEXPA6 overexpression Arabidopsis lines were much more resistant to cadmium stress. Our results revealed that the root system of the expa6 mutant was suppressed under cadmium stress, resulting in shorter plant height, reduced biomass, and a significant increase in cadmium content in the plants compared with wild-type plants, whereas LsEXPA6 overexpression lines had a well-developed root system and reduced cadmium accumulation in the roots and shoots of the plants. The above results indicated that overexpression of LsEXPA6 affected root development and reduced Cd absorption in Arabidopsis. In addition, the higher absorption capacity of nutrients, increased antioxidant enzymes activities, improved chlorophyll and photosynthetic function in the overexpression Arabidopsis plants, supported the Cd stress tolerance mechanism. Taken together, these results provided a new insight on the role of expansin proteins in the tolerance of plants to Cd stress by root cell elongation.

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Heavy metal (HM) toxicity has become a grave problem in the world since it leads to hazardous effects on living organisms. Transcriptomic/proteomic studies in plants have identified a large number of metal-responsive gene families. Of these, cytochrome-P450 (CYPs) family members are composed of enzymes carrying out detoxification of exogenous molecules. Here, we report a CYP-like protein encoded by Os08g01480 locus in rice that helps the plant to combat HM and other abiotic stresses. To functionally characterize CYP-like gene, cDNA and promoter were isolated from rice to develop Arabidopsis transgenic lines. Heterologous expression of Os08g01480 in Arabidopsis provided significant tolerance towards abiotic stresses. In silico analysis reveals that Os08g01480 might help plants to combat environmental stress via modulating auxin metabolism. Transgenic lines expressing reporter gene under control of Os08g01480 promoter demonstrated differential promoter activity in different tissues during environmental stresses. These studies indicated that differential expression of Os08g01480 might be modulating response of plants towards environmental stresses as well as in different developmental stages.

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Pyruvate is a central metabolite in cellular respiration and metabolism. It can neutralize reactive oxygen species (ROS), safeguard mitochondrial membrane potential, and regulate gene expression under oxidative stress. However, its role in abiotic stress tolerance in plants needs to be explored. Therefore, the current study investigated the role of pyruvate and its metabolism in response to different abiotic stresses in the model plant Arabidopsis thaliana. We retrieved transcript profiling data for pyruvate metabolism and transportation genes (D-LDH, AlaAT, PK, MPC, PDC, PDH, NAD-ME) from public databases. The study's findings indicate that these genes' expression is regulated in response to different abiotic stresses. Moreover, the promoter region of these genes contained multiple cis-acting elements like ABRE, ARE, P-box, and MBS, which are associated with plants' abiotic stress response. Furthermore, colorimetric analysis showed higher pyruvate content under different abiotic stresses. Therefore, exogenous pyruvate treatment was given before and after different abiotic stresses, which could combat the toxicity of pro-oxidant molecules by pyruvate intake. The semiquantitative RT-PCR analysis revealed that exogenous pyruvate treatment enhances the expression of important transcription factors WRKY2, GH3.3, DREB2A, and bZIP1, and stress-responsive genes e.g., APX1, ERD5, ADC2, and HSP70 in addition to abiotic stresses. Moreover, Arabidopsis plants pre-treated with pyruvate before oxidative stress showed less RBOHD expression. Additionally, pyruvate's cytoprotective role was compared to other well-known antioxidants such as NAC, Trolox, and GSH. Finally, untargeted GC-MS/MS analysis of abiotic stress-treated Arabidopsis plants showed a higher metabolite level of β-hydroxy-pyruvic acid, indicating the crucial role of pyruvate during abiotic stress.

Arabidopsis SIZ1 encodes a SUMO E3 ligase to regulate abiotic and biotic stress responses. Among SIZ1 or mammalian PIAS orthologs, plant SIZ1 proteins contain the plant homeodomain (PHD) finger, a C4HC3 zinc finger. Here, we investigated the importance of PHD of Arabidopsis SIZ1. The ProSIZ1::SIZ1(ΔPHD):GFP was unable to complement growth retardation, ABA hypersensitivity, and the cold-sensitive phenotype of the siz1 mutant, but ProSIZ1::SIZ1:GFP could. Substitution of C162S in the PHD finger was unable to complement the siz1 mutation. Tri-methylated histone H3K4 (H3K4me3) was recognized by PHD, not by PHD(C162S). WRKY70 was up-regulated in the siz1-2 mutant and H3K4me3 accumulated at high levels in the WRKY70 promoter. PHD interacts with ATX, which mediates methylation of histone, probably leading to suppression of ATX’s function. These results suggest that the PHD finger of SIZ1 is important for recognition of the histone code and is required for SIZ1 function and transcriptional suppression. Kenji Miura et al. investigate the role of the plant homeodomain (PHD) finger of the Arabidopsis SIZ1 protein. They show that the PHD finger is involved in hormone response and temperature sensitivity, and plays an important role in H3K4 methylation, thereby affecting recognition of histone code and transcriptional suppression.

Abstract N 6-methyladenosine (m6A) is the most abundant mRNA modification and plays diverse roles in eukaryotes, including plants. It regulates various processes, including plant growth, development, and responses to external or internal stress responses. However, the mechanisms underlying how m6A is related to environmental stresses in both mammals and plants remain elusive. Here, we identified EVOLUTIONARILY CONSERVED C-TERMINAL REGION 8 (ECT8) as an m6A reader protein and showed that its m6A-binding capability is required for salt stress responses in Arabidopsis (Arabidopsis thaliana). ECT8 accelerates the degradation of its target transcripts through direct interaction with the decapping protein DECAPPING 5 within processing bodies. We observed a significant increase in the ECT8 expression level under various environmental stresses. Using salt stress as a representative stressor, we found that the transcript and protein levels of ECT8 rise in response to salt stress. The increased abundance of ECT8 protein results in the enhanced binding capability to m6A-modified mRNAs, thereby accelerating their degradation, especially those of negative regulators of salt stress responses. Our results demonstrated that ECT8 acts as an abiotic stress sensor, facilitating mRNA decay, which is vital for maintaining transcriptome homeostasis and enhancing stress tolerance in plants. Our findings not only advance the understanding of epitranscriptomic gene regulation but also offer potential applications for breeding more resilient crops in the face of rapidly changing environmental conditions.

Background Small interfering RNAs (siRNAs) target homologous genomic DNA sequences for cytosine methylation, known as RNA-directed DNA methylation (RdDM), plays an important role in transposon control and regulation of gene expression in plants. Repressor of silencing 1 (ROS1) can negatively regulate the RdDM pathway. Results In this paper, we investigated the molecular mechanisms by which an upstream regulator ACD6 in the salicylic acid (SA) defense pathway, an ABA pathway-related gene ACO3 , and GSTF14 , an endogenous gene of the glutathione S-transferase superfamily, were induced by various abiotic stresses. The results demonstrated that abiotic stresses, including water deficit, cold, and salt stresses, induced demethylation of the repeats in the promoters of ACD6 , ACO3 , and GSTF14 and transcriptionally activated their expression. Furthermore, our results revealed that ROS1-mediated DNA demethylation plays an important role in the process of transcriptional activation of ACD6 and GSTF14 when Arabidopsis plants are subjected to cold stress. Conclusions This study revealed that ROS1 plays an important role in the molecular mechanisms associated with genes involved in defense pathways in response to abiotic stresses.

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Many plant genes have their expression modulated by stress conditions. Here, we used Arabidopsis FtsH5 protease, which expression is regulated by light stress, as bait in a yeast two-hybrid screen to search for new proteins involved in the stress response. As a result, we found FIP (FtsH5 Interacting Protein), which possesses an amino proximal cleavable transit peptide, a hydrophobic membrane-anchoring region, and a carboxyl proximal C4-type zinc-finger domain. In vivo experiments using FIP fused to green fluorescent protein (GFP) showed a plastid localization. This finding was corroborated by chloroplast import assays that showed FIP inserted in the thylakoid membrane. FIP expression was down-regulated in plants exposed to high light intensity, oxidative, salt, and osmotic stresses, whereas mutant plants expressing low levels of FIP were more tolerant to these abiotic stresses. Our data shows a new thylakoid-membrane protein involved with abiotic stress response in Arabidopsis thaliana.

Homeobox transcription factors are well known regulators of plant growth and development. In this study, we carried out functional analysis of two candidate stress-responsive HD-ZIP I class homeobox genes from rice, OsHOX22, and OsHOX24. These genes were highly up-regulated under various abiotic stress conditions at different stages of rice development, including seedling, mature and reproductive stages. The transcript levels of these genes were enhanced significantly in the presence of plant hormones, including abscisic acid (ABA), auxin, salicylic acid, and gibberellic acid. The recombinant full-length and truncated homeobox proteins were found to be localized in the nucleus. Electrophoretic mobility shift assay established the binding of these homeobox proteins with specific DNA sequences, AH1 (CAAT(A/T)ATTG) and AH2 (CAAT(C/G)ATTG). Transactivation assays in yeast revealed the transcriptional activation potential of full-length OsHOX22 and OsHOX24 proteins. Homo- and hetero-dimerization capabilities of these proteins have also been demonstrated. Further, we identified putative novel interacting proteins of OsHOX22 and OsHOX24 via yeast-two hybrid analysis. Over-expression of OsHOX24 imparted higher sensitivity to stress hormone, ABA, and abiotic stresses in the transgenic Arabidopsis plants as revealed by various physiological and phenotypic assays. Microarray analysis revealed differential expression of several stress-responsive genes in transgenic lines as compared to wild-type. Many of these genes were found to be involved in transcriptional regulation and various metabolic pathways. Altogether, our results suggest the possible role of OsHOX22/OsHOX24 homeobox proteins as negative regulators in abiotic stress responses.

Adverse environmental conditions are detrimental to plant growth and development. Acclimation to abiotic stress conditions involves activation of signaling pathways which often results in changes in gene expression via networks of transcription factors (TFs). Mediator is a highly conserved co-regulator complex and an essential component of the transcriptional machinery in eukaryotes. Some Mediator subunits have been implicated in stress-responsive signaling pathways; however, much remains unknown regarding the role of plant Mediator in abiotic stress responses. Here, we use RNA-seq to analyze the transcriptional response of Arabidopsis thaliana to heat, cold and salt stress conditions. We identify a set of common abiotic stress regulons and describe the sequential and combinatorial nature of TFs involved in their transcriptional regulation. Furthermore, we identify stress-specific roles for the Mediator subunits MED9, MED16, MED18 and CDK8, and putative TFs connecting them to different stress signaling pathways. Our data also indicate different modes of action for subunits or modules of Mediator at the same gene loci, including a co-repressor function for MED16 prior to stress. These results illuminate a poorly understood but important player in the transcriptional response of plants to abiotic stress and identify target genes and mechanisms as a prelude to further biochemical characterization.

No abstract available

N6-methyladenosine (m6A), the most abundant modification found in eukaryotic mRNAs, is interpreted by m6A "readers," thus playing a crucial role in regulating RNA metabolism. The YT521-B homology-domain (YTHD) proteins, also known as EVOLUTIONARILY CONSERVED C-TERMINAL REGION (ECT), are recognized as m6A reader proteins in plants and animals. Among the 13 potential YTHD family proteins in Arabidopsis thaliana, the functions of only a few members are known. In this study, we determined the function of ECT12 (YTH11) as a potential m6A reader that plays a crucial role in response to abiotic stresses. The loss-of-function ect12 mutants showed no noticeable developmental defects under normal conditions but displayed hypersensitivity to salt or dehydration stress. The salt- or dehydration-hypersensitive phenotypes were correlated with altered levels of several m6A-modified stress-responsive transcripts. Notably, the increased or decreased transcript levels were associated with each transcript's reduced or enhanced decay, respectively. Electrophoretic mobility shift and RNA-immunoprecipitation assays showed that ECT12 binds to m6A-modified RNAs both in vitro and in planta, suggesting its role as an m6A reader. Collectively, these results indicate that the potential m6A reader ECT12 regulates the stability of m6A-modified RNA transcripts, thereby facilitating the response of Arabidopsis to abiotic stresses.

B-cell lymphoma2 (Bcl-2)-associated athanogene (BAG) family proteins are evolutionary conserved across all eukaryotes. These proteins interact with HSP70/HSC70 and function as co-chaperones during stress response and developmental pathways. Compared to the animal counterpart, the BAG proteins in plants are much less studied and primarily Arabidopsis BAG proteins have been identified and characterized for their role in programmed cell death, homeostasis, growth and development, abiotic and biotic stress response. Here, we have identified BAG protein family (SlBAGs) in tomato, an economically important and a model fruit crop using genome-wide scanning. We have performed phylogenetic analysis, genes architecture assessment, chromosomal location and in silico promoter analysis. Our data suggest that SlBAGs show differential tissue specific expression pattern during plant development particularly fruit development and ripening. Furthermore, we reported that expression of SlBAGs is modulated during abiotic stresses and is regulated by stress hormones ABA and ethylene. In planta subcellular localization reveals their diverse subcellular localization, and many members are localized in nucleus and cytoplasm. Like previous reports, our protein–protein interaction network and yeast two-hybrid analysis uncover that SlBAGs interact with HSP70. The current study provides insights into role of SlBAGs in plant development particualry fruit ripening and abiotic stress response.

Dehydration response element binding (DREB) proteins are vital for plant abiotic stress responses, but the understanding of DREBs in bamboo, an important sustainable non-timber forest product, is limited. Here we conducted a comprehensive genome-wide analysis of the DREB gene family in Moso bamboo, representing the most important running bamboo species in Asia. In total, 44 PeDREBs were identified, and information on their gene structures, protein motifs, phylogenetic relationships, and stress-related cis-regulatory elements (CREs) was provided. Based on the bioinformatical analysis, we further analyzed PeDREBs from the A5 group and found that four of five PeDREB transcripts were induced by salt, drought, and cold stresses, and their proteins could bind to stress-related CREs. Among these, PeDREB28 was selected as a promising candidate for further functional characterization. PeDREB28 is localized in nucleus, has transcriptional activation activity, and could bind to the DRE- and coupling element 1- (CE1) CREs. Overexpression of PeDREB28 in Arabidopsis and bamboo improved plant abiotic stress tolerance. Transcriptomic analysis showed that broad changes due to the overexpression of PeDREB28. Furthermore, 628 genes that may act as the direct PeDREB28 downstream genes were identified by combining DAP-seq and RNA-seq analysis. Moreover, we confirmed that PeDREB28 could bind to the promoter of pyrabactin-resistance-like gene (DlaPYL3), which is a homolog of abscisic acid receptor in Arabidopsis, and activates its expression. In summary, our study provides important insights into the DREB gene family in Moso bamboo, and contributes to their functional verification and genetic engineering applications in the future.