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1.
Plant J ; 115(2): 434-451, 2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-37025007

RESUMO

Plant A/T-rich protein and zinc-binding protein (PLATZ) transcription factors play important roles in plant growth, development and abiotic stress responses. However, how PLATZ influences plant drought tolerance remains poorly understood. The present study showed that PLATZ4 increased drought tolerance in Arabidopsis thaliana by causing stomatal closure. Transcriptional profiling analysis revealed that PLATZ4 affected the expression of a set of genes involved in water and ion transport, antioxidant metabolism, small peptides and abscisic acid (ABA) signaling. Among these genes, the direct binding of PLATZ4 to the A/T-rich sequences in the plasma membrane intrinsic protein 2;8 (PIP2;8) promoter was identified. PIP2;8 consistently reduced drought tolerance in Arabidopsis through inhibiting stomatal closure. PIP2;8 was localized in the plasma membrane, exhibited water channel activity in Xenopus laevis oocytes and acted epistatically to PLATZ4 in regulating the drought stress response in Arabidopsis. PLATZ4 increased ABA sensitivity through upregulating the expression of ABSCISIC ACID INSENSITIVE 3 (ABI3), ABI4 and ABI5. The transcripts of PLATZ4 were induced to high levels in vegetative seedlings under drought and ABA treatments within 6 and 3 h, respectively. Collectively, these findings reveal that PLATZ4 positively influences plant drought tolerance through regulating the expression of PIP2;8 and genes involved in ABA signaling.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Ácido Abscísico/metabolismo , Resistência à Seca , Aquaporina 2/genética , Aquaporina 2/metabolismo , Plantas Geneticamente Modificadas/genética , Secas , Proteínas de Membrana/metabolismo , Membrana Celular/metabolismo , Regulação da Expressão Gênica de Plantas , Estresse Fisiológico/genética , Estômatos de Plantas/fisiologia
2.
Plant Mol Biol ; 114(3): 36, 2024 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-38598012

RESUMO

Increasing evidence indicates a strong correlation between the deposition of cuticular waxes and drought tolerance. However, the precise regulatory mechanism remains elusive. Here, we conducted a comprehensive transcriptome analysis of two wheat (Triticum aestivum) near-isogenic lines, the glaucous line G-JM38 rich in cuticular waxes and the non-glaucous line NG-JM31. We identified 85,143 protein-coding mRNAs, 4,485 lncRNAs, and 1,130 miRNAs. Using the lncRNA-miRNA-mRNA network and endogenous target mimic (eTM) prediction, we discovered that lncRNA35557 acted as an eTM for the miRNA tae-miR6206, effectively preventing tae-miR6206 from cleaving the NAC transcription factor gene TaNAC018. This lncRNA-miRNA interaction led to higher transcript abundance for TaNAC018 and enhanced drought-stress tolerance. Additionally, treatment with mannitol and abscisic acid (ABA) each influenced the levels of tae-miR6206, lncRNA35557, and TaNAC018 transcript. The ectopic expression of TaNAC018 in Arabidopsis also improved tolerance toward mannitol and ABA treatment, whereas knocking down TaNAC018 transcript levels via virus-induced gene silencing in wheat rendered seedlings more sensitive to mannitol stress. Our results indicate that lncRNA35557 functions as a competing endogenous RNA to modulate TaNAC018 expression by acting as a decoy target for tae-miR6206 in glaucous wheat, suggesting that non-coding RNA has important roles in the regulatory mechanisms responsible for wheat stress tolerance.


Assuntos
Arabidopsis , MicroRNAs , RNA Longo não Codificante , RNA Endógeno Competitivo , RNA Longo não Codificante/genética , Ácido Abscísico/farmacologia , Arabidopsis/genética , Manitol , MicroRNAs/genética , RNA Mensageiro , Triticum/genética , Ceras
3.
Plant Biotechnol J ; 22(8): 2093-2103, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38491985

RESUMO

Genetic transformation is a critical tool for gene editing and genetic improvement of plants. Although many model plants and crops can be genetically manipulated, genetic transformation systems for fruit trees are either lacking or perform poorly. We used Rhizobium rhizogenes to transfer the target gene into the hairy roots of Malus domestica and Actinidia chinensis. Transgenic roots were generated within 3 weeks, with a transgenic efficiency of 78.8%. Root to shoot conversion of transgenic hairy roots was achieved within 11 weeks, with a regeneration efficiency of 3.3%. Finally, the regulatory genes involved in stem cell activity were used to improve shoot regeneration efficiency. MdWOX5 exhibited the most significant effects, as it led to an improved regeneration efficiency of 20.6% and a reduced regeneration time of 9 weeks. Phenotypes of the overexpression of RUBY system mediated red roots and overexpression of MdRGF5 mediated longer root hairs were observed within 3 weeks, suggesting that the method can be used to quickly screen genes that influence root phenotype scores through root performance, such as root colour, root hair, and lateral root. Obtaining whole plants of the RUBY system and MdRGF5 overexpression lines highlights the convenience of this technology for studying gene functions in whole plants. Overall, we developed an optimized method to improve the transformation efficiency and stability of transformants in fruit trees.


Assuntos
Raízes de Plantas , Brotos de Planta , Plantas Geneticamente Modificadas , Transformação Genética , Plantas Geneticamente Modificadas/genética , Raízes de Plantas/genética , Raízes de Plantas/microbiologia , Raízes de Plantas/crescimento & desenvolvimento , Brotos de Planta/genética , Brotos de Planta/crescimento & desenvolvimento , Actinidia/genética , Actinidia/microbiologia , Malus/genética , Malus/microbiologia , Agrobacterium/genética , Árvores/genética
4.
PLoS Genet ; 17(11): e1009898, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34784357

RESUMO

Increasing evidence points to the tight relationship between alternative splicing (AS) and the salt stress response in plants. However, the mechanisms linking these two phenomena remain unclear. In this study, we have found that Salt-Responsive Alternatively Spliced gene 1 (SRAS1), encoding a RING-Type E3 ligase, generates two splicing variants: SRAS1.1 and SRAS1.2, which exhibit opposing responses to salt stress. The salt stress-responsive AS event resulted in greater accumulation of SRAS1.1 and a lower level of SRAS1.2. Comprehensive phenotype analysis showed that overexpression of SRAS1.1 made the plants more tolerant to salt stress, whereas overexpression of SRAS1.2 made them more sensitive. In addition, we successfully identified the COP9 signalosome 5A (CSN5A) as the target of SRAS1. CSN5A is an essential player in the regulation of plant development and stress. The full-length SRAS1.1 promoted degradation of CSN5A by the 26S proteasome. By contrast, SRAS1.2 protected CSN5A by competing with SRAS1.1 on the same binding site. Thus, the salt stress-triggered AS controls the ratio of SRAS1.1/SRAS1.2 and switches on and off the degradation of CSN5A to balance the plant development and salt tolerance. Together, these results provide insights that salt-responsive AS acts as post-transcriptional regulation in mediating the function of E3 ligase.


Assuntos
Processamento Alternativo , Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiologia , Complexo do Signalossomo COP9/genética , Estresse Salino , Ubiquitina-Proteína Ligases/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Genes de Plantas , Isoformas de Proteínas/genética , Salinidade , Ubiquitina-Proteína Ligases/genética
5.
New Phytol ; 240(2): 710-726, 2023 10.
Artigo em Inglês | MEDLINE | ID: mdl-37547968

RESUMO

MicroRNAs (miRNAs) play crucial roles in regulating plant development and stress responses. However, the functions and mechanism of intronic miRNAs in plants are poorly understood. This study reports a stress-responsive RNA splicing mechanism for intronic miR400 production, whereby miR400 modulates reactive oxygen species (ROS) accumulation and improves plant tolerance by downregulating its target expression. To monitor the intron splicing events, we used an intronic miR400 splicing-dependent luciferase transgenic line. Luciferase activity was observed to decrease after high cadmium concentration treatment due to the retention of the miR400-containing intron, which inhibited the production of mature miR400. Furthermore, we demonstrated that under Cd treatments, Pentatricopeptide Repeat Protein 1 (PPR1), the target of miR400, acts as a positive regulator by inducing ROS accumulation. Ppr1 mutation affected the Complex III activity in the electron transport chain and RNA editing of the mitochondrial gene ccmB. This study illustrates intron splicing as a key step in intronic miR400 production and highlights the function of intronic miRNAs as a 'signal transducer' in enhancing plant stress tolerance.


Assuntos
Arabidopsis , MicroRNAs , MicroRNAs/genética , MicroRNAs/metabolismo , Arabidopsis/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Íntrons/genética , Splicing de RNA/genética , Regulação da Expressão Gênica de Plantas
6.
Int J Mol Sci ; 24(2)2023 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-36674568

RESUMO

Salt stress is one of the major environmental threats to plant growth and development. However, the mechanisms of plants responding to salt stress are not fully understood. Through genetic screening, we identified and characterized a salt-sensitive mutant, ses5 (sensitive to salt 5), in Arabidopsis thaliana. Positional cloning revealed that the decreased salt-tolerance of ses5 was caused by a mutation in the transcription factor BP (BREVIPEDICELLUS). BP regulates various developmental processes in plants. However, the biological function of BP in abiotic stress-signaling and tolerance are still not clear. Compared with wild-type plants, the bp mutant exhibited a much shorter primary-root and lower survival rate under salt treatment, while the BP overexpressors were more tolerant. Further analysis showed that BP could directly bind to the promoter of XTH7 (xyloglucan endotransglucosylase/hydrolase 7) and activate its expression. Resembling the bp mutant, the disruption of XTH7 gave rise to salt sensitivity. These results uncovered novel roles of BP in positively modulating salt-stress tolerance, and illustrated a putative working mechanism.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Proteínas de Homeodomínio , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Regulação da Expressão Gênica de Plantas , Plantas Geneticamente Modificadas/genética , Regiões Promotoras Genéticas , Tolerância ao Sal/genética , Estresse Fisiológico/genética , Proteínas de Homeodomínio/metabolismo
7.
J Integr Plant Biol ; 65(8): 1846-1851, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37052306

RESUMO

Phytohormone abscisic acid (ABA) plays vital roles in stress tolerance, while long-term overactivation of ABA signaling suppresses plant growth and development. However, the braking mechanism of ABA responses is not clear. Protein tyrosine sulfation catalyzed by tyrosylprotein sulfotransferase (TPST) is a critical post-translational modification. Through genetic screening, we identified a tpst mutant in Arabidopsis that was hypersensitive to ABA. In-depth analysis revealed that TPST could interact with and sulfate SnRK2.2/2.3/2.6, which accelerated their degradation and weakened the ABA signaling. Taken together, these findings uncovered a novel mechanism of desensitizing ABA responses via protein sulfation.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Ácido Abscísico/farmacologia , Ácido Abscísico/metabolismo , Sulfotransferases/genética , Sulfotransferases/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo
8.
BMC Genomics ; 23(1): 145, 2022 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-35180840

RESUMO

BACKGROUNDS: Lysine 2-hydroxyisobutyrylation (Khib) is a newly discovered posttranslational modification (PTM) and has been identified in several prokaryotic and eukaryotic organisms. Fusarium graminearum, a major pathogen of Fusarium head blight (FHB) in cereal crops, can cause considerable yield loss and produce various mycotoxins that threaten human health. The application of chemical fungicides such as tebuconazole (TEC) remains the major method to control this pathogen. However, the distribution of Khib in F. graminearum and whether Khib is remodified in response to fungicide stress remain unknown. RESULTS: Here, we carried out a proteome-wide analysis of Khib in F. graminearum, identifying the reshaping of the lysine 2-hydroxyisobutyrylome by tebuconazole, using the most recently developed high-resolution LC-MS/MS technique in combination with high-specific affinity enrichment. Specifically, 3501 Khib sites on 1049 proteins were identified, and 1083 Khib sites on 556 modified proteins normalized to the total protein content were changed significantly after TEC treatment. Bioinformatics analysis showed that Khib proteins are involved in a wide range of biological processes and may be involved in virulence and deoxynivalenol (DON) production, as well as sterol biosynthesis, in F. graminearum. CONCLUSIONS: Here, we provided a wealth of resources for further study of the roles of Khib in the fungicide resistance of F. graminearum. The results enhanced our understanding of this PTM in filamentous ascomycete fungi and provided insight into the remodification of Khib sites during azole fungicide challenge in F. graminearum.


Assuntos
Fusarium , Cromatografia Líquida , Humanos , Lisina/metabolismo , Doenças das Plantas , Proteômica , Espectrometria de Massas em Tandem , Triazóis
9.
J Exp Bot ; 73(18): 6417-6433, 2022 10 18.
Artigo em Inglês | MEDLINE | ID: mdl-35709944

RESUMO

Increasing temperature is one of the major threats to maize growth and yield globally. Under heat stress conditions, intracellular protein homeostasis is seriously disturbed, leading to accumulation of abnormally folded proteins, especially in the endoplasmic reticulum (ER). Molecular chaperones are vital players in the renaturation process and in preventing protein aggregation. However, heat stress tolerance-associated chaperones are not well documented in maize. Here, we characterized the biological roles of HEAT UP-REGULATED GENE 1 (ZmHUG1) in maize. ZmHUG1 encodes a heat-inducible holdase-type molecular chaperone localized in the ER. Knockout mutant of ZmHUG1 exhibited remarkably enhanced sensitivity to heat stress. Accordingly, the zmhug1 mutant showed severe ER stress under high temperature. MAIZE PRENYLATED RAB ACCEPTOR 1.C1 (ZmPRA1.C1) was identified as a client of ZmHUG1, and heat-induced aggregation of ZmPRA1.C1 was accelerated in the zmhug1 mutant. Furthermore, the expression of ZmHUG1 was rapidly transactivated by ER stress sensor BASIC LEUCINE ZIPPER DOMAIN 60 (bZIP60) when heat stress occurred. This study reveals a ZmHUG1-based thermo-protective mechanism in maize.


Assuntos
Agregados Proteicos , Termotolerância , Resposta ao Choque Térmico , Termotolerância/genética , Retículo Endoplasmático/metabolismo , Estresse do Retículo Endoplasmático , Chaperonas Moleculares/metabolismo
10.
J Integr Plant Biol ; 64(12): 2344-2360, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36223079

RESUMO

Active DNA demethylation effectively modulates gene expression during plant development and in response to stress. However, little is known about the upstream regulatory factors that regulate DNA demethylation. We determined that the demethylation regulator 1 (demr1) mutant exhibits a distinct DNA methylation profile at selected loci queried by methylation-sensitive polymerase chain reaction and globally based on whole-genome bisulfite sequencing. Notably, the transcript levels of the DNA demethylase gene REPRESSOR OF SILENCING 1 (ROS1) were lower in the demr1 mutant. We established that DEMR1 directly binds to the ROS1 promoter in vivo and in vitro, and the methylation level in the DNA methylation monitoring sequence of ROS1 promoter decreased by 60% in the demr1 mutant. About 40% of the hyper-differentially methylated regions (DMRs) in the demr1 mutant were shared with the ros1-4 mutant. Genetic analysis indicated that DEMR1 acts upstream of ROS1 to positively regulate abscisic acid (ABA) signaling during seed germination and seedling establishment stages. Surprisingly, the loss of DEMR1 function also caused a rise in methylation levels of the mitochondrial genome, impaired mitochondrial structure and an early flowering phenotype. Together, our results show that DEMR1 is a novel regulator of DNA demethylation of both the nuclear and mitochondrial genomes in response to ABA and plant development in Arabidopsis.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Genoma Mitocondrial , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Genoma Mitocondrial/genética , Proteínas Tirosina Quinases/genética , Proteínas Tirosina Quinases/metabolismo , Desmetilação do DNA , Proteínas Nucleares/metabolismo , Proteínas Proto-Oncogênicas/genética , Arabidopsis/genética , Arabidopsis/metabolismo , Metilação de DNA/genética , Regulação da Expressão Gênica de Plantas/genética
11.
New Phytol ; 230(2): 641-655, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33421141

RESUMO

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 serine/arginine-rich (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 messenger RNA (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.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Processamento Alternativo/genética , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Arginina , Regulação da Expressão Gênica de Plantas , Fatores de Processamento de RNA , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Serina
12.
J Exp Bot ; 72(18): 6260-6273, 2021 09 30.
Artigo em Inglês | MEDLINE | ID: mdl-34097059

RESUMO

C-terminally encoded peptides (CEPs) are small peptides, typically post-translationally modified, and highly conserved in many species. CEPs are known to inhibit plant growth and development, but the mechanisms are not well understood. In this study, 14 CEPs were identified in Setaria italica and divided into two groups. The transcripts of most SiCEPs were more abundant in roots than in other detected tissues. SiCEP3, SiCEP4, and SiCEP5 were also highly expressed in panicles. Moreover, expression of all SiCEPs was induced by abiotic stresses and phytohormones. SiCEP3 overexpression and application of synthetic SiCEP3 both inhibited seedling growth. In the presence of abscisic acid (ABA), growth inhibition and ABA content in seedlings increased with the concentration of SiCEP3. Transcripts encoding eight ABA transporters and six ABA receptors were induced or repressed by synthetic SiCEP3, ABA, and their combination. Further analysis using loss-of-function mutants of Arabidopsis genes functioning as ABA transporters, receptors, and in the biosynthesis and degradation of ABA revealed that SiCEP3 promoted ABA import at least via NRT1.2 (NITRATE TRANSPORTER 1.2) and ABCG40 (ATP-BINDING CASSETTE G40). In addition, SiCEP3, ABA, or their combination inhibited the kinase activities of CEP receptors AtCEPR1/2. Taken together, our results indicated that the CEP-CEPR module mediates ABA signaling by regulating ABA transporters and ABA receptors in planta.


Assuntos
Proteínas de Arabidopsis , Setaria (Planta) , Ácido Abscísico , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Regulação da Expressão Gênica de Plantas , Peptídeos , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
13.
Phytopathology ; 111(8): 1420-1427, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-33399013

RESUMO

Fusarium graminearum is one of the most important causal agents of Fusarium head blight disease and is controlled mainly by chemicals such as demethylation inhibitor (DMI) fungicides. FgCYP51B is one of the DMI targets in F. graminearum, and Tyrosine123 (Y123) is an important amino acid in F. graminearum CYP51B, located in one of predicted substrate binding pockets based on the binding mode between DMIs and CYP51B. Previous studies suggest that resistance to DMI fungicides is attributed primarily to point mutations in the CYP51 gene and that the Y123H mutation in F. verticillioides CYP51 confers prochloraz resistance in the laboratory. To investigate the function of FgCYP51B Y123 residue in the growth and development, pathogenicity, and DMI resistance, we generated and analyzed the FgCYP51B Y123H mutant. Results revealed that the Y123H mutation led to reduced conidial sporulation and affected ascospore development; moreover, the mutation conferred reduced sensitivity to prochloraz. Quantitative PCR and molecular docking were performed to investigate the resistance mechanism. Results indicated that Y123H mutation changed the target gene expression and decreased the binding affinity of FgCYP51 to prochloraz. These results will attract more attention to the potential DMI-resistant mutation of F. graminearum and increase our understanding of the DMI resistance mechanism.


Assuntos
Fungicidas Industriais , Fusarium , Farmacorresistência Fúngica/genética , Fungicidas Industriais/farmacologia , Fusarium/efeitos dos fármacos , Fusarium/genética , Imidazóis , Simulação de Acoplamento Molecular , Mutação , Doenças das Plantas/microbiologia
14.
J Integr Plant Biol ; 63(3): 484-493, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32970364

RESUMO

Mitogen activated protein kinase kinase kinase 18 (MAPKKK18) mediated signaling cascade plays important roles in Arabidopsis drought stress tolerance. However, the post-translational modulation patterns of MAPKKK18 are not characterized. In this study, we found that the protein level of MAPKKK18 was tightly controlled by the 26S proteasome. Ubiquitin ligases RGLG1 and RGLG2 ubiquitinated MAPKKK18 at lysine residue K32 and K154, and promoted its degradation. Deletion of RGLG1 and RGLG2 stabilized MAPKKK18 and further enhanced the drought stress tolerance of MAPKKK18-overexpression plants. Our data demonstrate that RGLG1 and RGLG2 negatively regulate MAPKKK18-mediated drought stress tolerance in Arabidopsis.


Assuntos
Adaptação Fisiológica , Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiologia , Secas , MAP Quinase Quinase Quinases/metabolismo , Domínios RING Finger , Estresse Fisiológico , Ubiquitina-Proteína Ligases/metabolismo , Lisina/metabolismo , Mutação/genética , Complexo de Endopeptidases do Proteassoma/metabolismo , Ligação Proteica , Proteólise , Ubiquitinação
15.
J Exp Bot ; 71(18): 5589-5602, 2020 09 19.
Artigo em Inglês | MEDLINE | ID: mdl-32453821

RESUMO

Although the salt overly sensitive (SOS) pathway plays essential roles in conferring salt tolerance in Arabidopsis thaliana, the regulatory mechanism underlying SOS gene expression remains largely unclear. In this study, AtPLATZ2 was found to function as a direct transcriptional suppressor of CBL4/SOS3 and CBL10/SCaBP8 in the Arabidopsis salt stress response. Compared with wild-type plants, transgenic plants constitutively overexpressing AtPLATZ2 exhibited increased sensitivity to salt stress. Loss of function of PLATZ2 had no observed salt stress phenotype in Arabidopsis, while the double mutant of PLATZ2 and PLATZ7 led to weaker salt stress tolerance than wild-type plants. Overexpression of AtPLATZ2 in transgenic plants decreased the expression of CBL4/SOS3 and CBL10/SCaBP8 under both normal and saline conditions. AtPLATZ2 directly bound to A/T-rich sequences in the CBL4/SOS3 and CBL10/SCaBP8 promoters in vitro and in vivo, and inhibited CBL4/SOS3 promoter activity in the plant leaves. The salt sensitivity of #11 plants constitutively overexpressing AtPLATZ2 was restored by the overexpression of CBL4/SOS3 and CBL10/SCaBP8. Salt stress-induced Na+ accumulation in both the shoots and roots was more exaggerated in AtPLATZ2-overexpressing plants than in the wild type. The salt stress-induced Na+ accumulation in #11 seedlings was also rescued by the overexpression of CBL4/SOS3 and CBL10/SCaBP8. Furthermore, the transcription of AtPLATZ2 was induced in response to salt stress. Collectively, these results suggest that AtPLATZ2 suppresses plant salt tolerance by directly inhibiting CBL4/SOS3 and CBL10/SCaBP8, and functions redundantly with PLATZ7.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Ligação ao Cálcio/metabolismo , Regulação da Expressão Gênica de Plantas , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Tolerância ao Sal/genética , Plântula/genética , Plântula/metabolismo
16.
Mol Cell ; 48(4): 521-31, 2012 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-23063528

RESUMO

MicroRNAs (miRNAs) have emerged as a class of regulators of gene expression through posttranscriptional degradation or translational repression in living cells. Increasing evidence points to the important relationship between miRNAs and environmental stress responses, but the regulatory mechanisms in plants are poorly understood. Here, we found that Arabidopsis thaliana intronic miR400 was cotranscribed with its host gene (At1g32583) and downregulated by heat treatment. Intriguingly, an alternative splicing (AS) event that occurred in the intron (306 bp) where MIR400 was located was specifically induced by heat stress. A 100 bp fragment was excised, and the remaining 206 bp intron containing MIR400 transcripts was retained in the host gene. The stress-induced AS event thus resulted in greater accumulation of miR400 primary transcripts and a low level of mature miR400. Together, these results provide the direct evidence that AS acts as a regulatory mechanism linking miRNAs and environmental stress in plants.


Assuntos
Processamento Alternativo , Arabidopsis/genética , Arabidopsis/metabolismo , Temperatura Alta , MicroRNAs/metabolismo , Processamento Pós-Transcricional do RNA , Estresse Fisiológico/genética , Processamento Alternativo/genética , Arabidopsis/citologia , Íntrons , MicroRNAs/genética , Transcrição Gênica
17.
J Integr Plant Biol ; 62(4): 403-420, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31001913

RESUMO

Low molecular weight secreted peptides have recently been shown to affect multiple aspects of plant growth, development, and defense responses. Here, we performed stepwise BLAST filtering to identify unannotated peptides from the Arabidopsis thaliana protein database and uncovered a novel secreted peptide family, secreted transmembrane peptides (STMPs). These low molecular weight peptides, which consist of an N-terminal signal peptide and a transmembrane domain, were primarily localized to extracellular compartments but were also detected in the endomembrane system of the secretory pathway, including the endoplasmic reticulum and Golgi. Comprehensive bioinformatics analysis identified 10 STMP family members that are specific to the Brassicaceae family. Brassicaceae plants showed dramatically inhibited root growth upon exposure to chemically synthesized STMP1 and STMP2. Arabidopsis overexpressing STMP1, 2, 4, 6, or 10 exhibited severely arrested growth, suggesting that STMPs are involved in regulating plant growth and development. In addition, in vitro bioassays demonstrated that STMP1, STMP2, and STMP10 have antibacterial effects against Pseudomonas syringae pv. tomato DC3000, Ralstonia solanacearum, Bacillus subtilis, and Agrobacterium tumefaciens, demonstrating that STMPs are antimicrobial peptides. These findings suggest that STMP family members play important roles in various developmental events and pathogen defense responses in Brassicaceae plants.


Assuntos
Brassicaceae/imunologia , Brassicaceae/microbiologia , Peptídeos/metabolismo , Desenvolvimento Vegetal , Sequência de Aminoácidos , Arabidopsis/metabolismo , Bactérias/efeitos dos fármacos , Brassicaceae/genética , Brassicaceae/crescimento & desenvolvimento , Cromossomos de Plantas/genética , Regulação da Expressão Gênica de Plantas , Anotação de Sequência Molecular , Peptídeos/química , Peptídeos/farmacologia , Fenótipo , Filogenia , Regiões Promotoras Genéticas/genética , Especificidade da Espécie , Estresse Fisiológico/genética , Frações Subcelulares/metabolismo
18.
Proteins ; 87(4): 257-264, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30561148

RESUMO

MoSub1 is an ortholog of yeast single stranded DNA binding protein Sub1 or human PC4 from rice blast fungus. All of them share a similar DNA binding region and may have similar biological roles. The well-studied Sub1/PC4 has been reported to play multiple roles in DNA metabolic processes, such as transcription and DNA repair and their DNA binding capacity is significantly affected by phosphorylation. Here, we determined the crystal structure of MoSub1 complexed with ssDNA in a phosphate solution. The crystal structure of the MoSub1-ssDNA complex was solved to a resolution of 2.04 Å. A phosphate ion at the interface of the protein-DNA interaction of the complex bridged the lys84 of the protein and two nucleotides. The DNA was bound in novel mode (L mode) in the MoSub1 complex in the presence of phosphate ions, while DNA bound in the straight mode in the absence of the phosphate ion and in U mode in the same binding motif of the PC4-ssDNA complex. The crystal structure of the complex and a small-angle X-ray scattering analysis revealed that the phosphate ion at the protein-DNA interface affected the DNA binding mode of MoSub1 to oligo-DNA and provided a new structural clue for studying its functions.


Assuntos
DNA de Cadeia Simples/metabolismo , Proteínas de Ligação a DNA/metabolismo , Proteínas Fúngicas/metabolismo , Magnaporthe/metabolismo , Sequência de Aminoácidos , Sítios de Ligação , Soluções Tampão , Cristalografia por Raios X , DNA de Cadeia Simples/química , Proteínas de Ligação a DNA/química , Proteínas Fúngicas/química , Magnaporthe/química , Modelos Moleculares , Fosfatos/química , Fosfatos/metabolismo , Ligação Proteica , Conformação Proteica
19.
Plant Mol Biol ; 99(4-5): 395-406, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30701352

RESUMO

KEY MESSAGE: CYSTM3, a small mitochondrial protein, acts as a negative regulator in salt stress response by preventing Na+ efflux and disturbing reactive oxygen species (ROS) homeostasis in Arabidopsis. Cysteine-rich transmembrane module (CYSTM) is a not well characterized small peptide family in plants. In this study, we identified a novel mitochondrion-localized CYSTM member CYSTM3 from Arabidopsis, which was ubiquitously expressed in different tissues and dramatically induced by salt stress. Transgenic plants overexpressing CYSTM3 (OE) displayed hypersensitivity to salt stress compared with wild type (WT) plants, whereas a knockout mutant cystm3 was more tolerant to high salinity than WT. Moreover, OE lines accumulated higher contents of Na+ and ROS than WT and cystm3 upon exposure to high salinity. Further analysis revealed that CYSTM3 could deter root Na+ efflux and inhibit the activities of a range of ROS scavenging enzymes in Arabidopsis. In addition, the transcripts of nuclear salt stress-responsive genes were over-activated in cystm3 than those in WT and OE lines. Taken together, Arabidopsis CYSTM3 acts as a negative regulator in salt stress tolerance.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Estresse Salino , Tolerância ao Sal/fisiologia , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Clorofila/análise , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Técnicas de Inativação de Genes , Homeostase , Proteínas Mitocondriais/genética , Proteínas Mitocondriais/metabolismo , Estresse Oxidativo , Plantas Geneticamente Modificadas , Espécies Reativas de Oxigênio , Salinidade , Tolerância ao Sal/genética , Sódio/metabolismo
20.
Biochem Biophys Res Commun ; 513(3): 582-588, 2019 06 04.
Artigo em Inglês | MEDLINE | ID: mdl-30981503

RESUMO

Heat stress significantly disturbs the protein folding and processing capability in plants. Molecular chaperones are vital players in unfolded/misfolded protein assembly and abiotic stress tolerance. Here, we reported SES1, which encodes an endoplasmic reticulum (ER) localized molecular chaperone, is required for Arabidopsis heat tolerance. SES1 is obviously induced by heat treatment and ses1 mutants are hypersensitive to heat stress. The unfolded protein response genes were up-regulated, while cytosolic protein response genes were down-regulated in ses1 after heat stress. Furthermore, ER stress sensor basic leucine zipper 28 (bZIP28) acts as the upstream transcriptional activator of SES1 by binding to its promoter region. These results provide new insights into heat stress responses and ER stress, and shed lights on the mechanism of SES1 in modulating heat resistance.


Assuntos
Proteínas de Arabidopsis/fisiologia , Arabidopsis/genética , Chaperonas Moleculares/fisiologia , Termotolerância/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/biossíntese , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Regulação da Expressão Gênica de Plantas , Chaperonas Moleculares/biossíntese , Chaperonas Moleculares/genética , Ativação Transcricional , Resposta a Proteínas não Dobradas
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