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1.
Ecotoxicol Environ Saf ; 202: 110918, 2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-32800253

RESUMO

This work evaluates different generations of transgenic (cp4-EPSPS gene) and non-transgenic soybean plants through proteomics and metabolomics. For proteomics purpose, 24 differentially abundant protein spots were found through 2-D DIGE, being 4 belonging to transgenic plants. From this total, 19 were successfully identified, storage proteins as predominant class. Some identified proteins are involved in growing and cell division, and stress response, such as LEA and dehydrin. For metabolomics, 17 compounds were putatively annotated, mainly belonging to the secondary metabolism, such as flavonoids. From these analyzes, all generations and varieties of the soybean are prone to be differentiate by PLS-DA. According to our results, transgenic plants appear to be more stable than non-transgenic ones. In addition, the omics-based approaches allowed access some relations between those differential spot proteins and metabolites, mainly those storage proteins and flavonoid.


Assuntos
Plantas Geneticamente Modificadas/fisiologia , Soja/fisiologia , Metabolômica , Plantas Geneticamente Modificadas/metabolismo , Proteômica , Sementes/metabolismo , Soja/genética , Soja/metabolismo
2.
Nat Commun ; 11(1): 3439, 2020 07 10.
Artigo em Inglês | MEDLINE | ID: mdl-32651385

RESUMO

Various stress conditions induce the nuclear translocation of cytosolic glyceraldehyde-3-phosphate dehydrogenase (GAPC), but its nuclear function in plant stress responses remains elusive. Here we show that GAPC interacts with a transcription factor to promote the expression of heat-inducible genes and heat tolerance in Arabidopsis. GAPC accumulates in the nucleus under heat stress. Overexpression of GAPC enhances heat tolerance of seedlings and the expression of heat-inducible genes whereas knockout of GAPCs has opposite effects. Screening of Arabidopsis transcription factors identifies nuclear factor Y subunit C10 (NF-YC10) as a GAPC-binding protein. The effects of GAPC overexpression are abolished when NF-YC10 is deficient, the heat-induced nuclear accumulation of GAPC is suppressed, or the GAPC-NF-YC10 interaction is disrupted. GAPC overexpression also enhances the binding ability of NF-YC10 to its target promoter. The results reveal a cellular and molecular mechanism for the nuclear moonlighting of a glycolytic enzyme in plant response to environmental changes.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Arabidopsis/fisiologia , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Fator de Ligação a CCAAT/genética , Fator de Ligação a CCAAT/metabolismo , Núcleo Celular/genética , Núcleo Celular/metabolismo , Núcleo Celular/fisiologia , Citosol/metabolismo , Citosol/fisiologia , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Gliceraldeído-3-Fosfato Desidrogenases/genética , Resposta ao Choque Térmico/genética , Resposta ao Choque Térmico/fisiologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Plântula/genética , Plântula/metabolismo , Plântula/fisiologia , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
3.
PLoS Genet ; 16(5): e1008807, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32407354

RESUMO

Pollen wall consists of several complex layers which form elaborate species-specific patterns. In Arabidopsis, the transcription factor ABORTED MICROSPORE (AMS) is a master regulator of exine formation, and another transcription factor, TRANSPOSABLE ELEMENT SILENCING VIA AT-HOOK (TEK), specifies formation of the nexine layer. However, knowledge regarding the temporal regulatory roles of TEK in pollen wall development is limited. Here, TEK-GFP driven by the AMS promoter was prematurely expressed in the tapetal nuclei, leading to complete male sterility in the pAMS:TEK-GFP (pat) transgenic lines with the wild-type background. Cytological observations in the pat anthers showed impaired callose synthesis and aberrant exine patterning. CALLOSE SYNTHASE5 (CalS5) is required for callose synthesis, and expression of CalS5 in pat plants was significantly reduced. We demonstrated that TEK negatively regulates CalS5 expression after the tetrad stage in wild-type anthers and further discovered that premature TEK-GFP in pat directly represses CalS5 expression through histone modification. Our findings show that TEK flexibly mediates its different functions via different temporal regulation, revealing that the temporal regulation of TEK is essential for exine patterning. Moreover, the result that the repression of CalS5 by TEK after the tetrad stage coincides with the timing of callose wall dissolution suggests that tapetum utilizes temporal regulation of genes to stop callose wall synthesis, which, together with the activation of callase activity, achieves microspore release and pollen wall patterning.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Pólen/fisiologia , Fatores de Transcrição/metabolismo , Arabidopsis/fisiologia , Proteínas de Arabidopsis/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Epigênese Genética , Regulação da Expressão Gênica de Plantas , Glucosiltransferases/metabolismo , Histonas/metabolismo , Metilação , Plantas Geneticamente Modificadas/fisiologia , Pólen/genética , Regiões Promotoras Genéticas
4.
Plant Mol Biol ; 103(6): 689-704, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32472480

RESUMO

KEY MESSAGE: CmCAD2 and CmCAD3 function more positively than CmCAD1 in oriental melon for lignin synthesis which is important to ensure internal water status and thus for drought tolerance. Well-lignification may be the guarantee of efficient axial water transport and barrier of lateral water flow in oriental melon tolerating drought stress, however remains to be verified. As an important enzyme in monolignol synthesis pathway, five cinnamyl alcohol dehydrogenase (CAD) genes were generally induced in melon seedlings by drought. Here we further revealed the roles of CmCAD1, 2, and 3 in lignin synthesis and for drought tolerance. Results found that overexpressing CmCAD2 or 3 strongly recovered CAD activities, lignin synthesis and composition in Arabidopsis cadc cadd, whose lignin synthesis is disrupted, while CmCAD1 functioned modestly. In melon seedlings, silenced CmCAD2 and 3 individually or collectively decreased CAD activities and lignin depositions drastically, resulting in dwarfed phenotypes. Reduced lignin, mainly composed by guaiacyl units catalyzed by CmCAD3, is mainly due to the limited lignification in tracheary elements and development of Casparion strip. While CmCAD1 and 2 exhibited catalysis to p-coumaraldehyde and sinapaldehyde, respectively. Compared with CmCAD1, drought treatments revealed higher sensitivity of CmCAD2 and/or 3 silenced melon seedlings, accompanying with lower relative water contents, water potentials and relatively higher total soluble sugar contents. Slightly up-regulated expressions of aquaporin genes together with limited lignification might imply higher lateral water loss in stems of silenced lines. In Arabidopsis, CmCAD2 and 3 transgenic lines enhanced cadc cadd drought tolerance through recovering lignin synthesis and root development, accompanying with decreased electrolyte leakage ratios and increased RWCs, thus improved survival rates. Briefly, lignin synthesized by CmCAD2 and 3 functions importantly for drought tolerance in melon.


Assuntos
Plântula/metabolismo , Cucumis melo/metabolismo , Cucumis melo/fisiologia , Secas , Regulação da Expressão Gênica de Plantas/fisiologia , Lignina/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Plântula/fisiologia
5.
Artigo em Inglês | MEDLINE | ID: mdl-32229406

RESUMO

NAC [No apical meristem (NAM), Arabidopsis transcription activation factor (ATAF), Cup-shaped cotyledon (CUC)] transcription factors (TFs) play an important role in plant growth and response to various environmental stress. Drought stress is the primary factor limiting the growth and fruit quality of grapevines worldwide. However, the biological function of the NAC family members in grapevine is not clear. In this study, we reported that VvNAC08, a novel NAC transcription factor gene, was expressed after drought, salicylic acid (SA) and abscisic acid (ABA), jasmonic acid (JA) and melatonin (MT) treatments in grapevine. VvNAC08 was expressed in various tissues. The open reading frame (ORF) of VvNAC08 was 792 bp, encoding 263 amino acids. The VvNAC08 protein could bind to NACRS [CGTA/CACG] in yeast. When subjected to drought and dehydration stress, VvNAC08-overexpression (OE) Arabidopsis had a higher survival rate and a lower water loss rate than wild type (WT) plants. Under drought conditions, transgenic Arabidopsis overexpressing VvNAC08 had a lower malondialdehyde (MDA), H2O2 contents, but a higher peroxidase (POD), superoxide dismutase (SOD) and catalase (CAT) activities as well as more proline content. Moreover, the expressions of marker genes, including ABI5, AREB1, COR15A, COR47, P5CS, RD22, and RD29A, were up-regulated in VvNAC08-overexpression lines when subjected to drought treatments. The results suggest that the transgenic Arabidopsis overexpressing VvNAC08 enhances resistance to drought while up-regulating the expressions of ABA- and stress-related genes.


Assuntos
Arabidopsis/fisiologia , Secas , Proteínas de Plantas/genética , Estresse Fisiológico , Fatores de Transcrição/genética , Vitis/genética , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Plantas Geneticamente Modificadas/fisiologia
6.
Science ; 367(6485): 1482-1485, 2020 03 27.
Artigo em Inglês | MEDLINE | ID: mdl-32217727

RESUMO

The premature abscission of flowers and fruits limits crop yield under environmental stress. Drought-induced flower drop in tomato plants was found to be regulated by phytosulfokine (PSK), a peptide hormone previously known for its growth-promoting and immune-modulating activities. PSK formation in response to drought stress depends on phytaspase 2, a subtilisin-like protease of the phytaspase subtype that generates the peptide hormone by aspartate-specific processing of the PSK precursor in the tomato flower pedicel. The mature peptide acts in the abscission zone where it induces expression of cell wall hydrolases that execute the abscission process. Our results provide insight into the molecular control of abscission as regulated by proteolytic processing to generate a small plant peptide hormone.


Assuntos
Secas , Flores/fisiologia , Lycopersicon esculentum/fisiologia , Hormônios Peptídicos/fisiologia , Proteínas de Plantas/fisiologia , Estresse Fisiológico , Parede Celular/enzimologia , Etilenos , Frutas/fisiologia , Técnicas de Silenciamento de Genes , Hidrolases/fisiologia , Ácidos Indolacéticos , Peptídeos , Plantas Geneticamente Modificadas/fisiologia , Transdução de Sinais
7.
Artigo em Inglês | MEDLINE | ID: mdl-32114414

RESUMO

Heat shock transcription factors (Hsfs) play an important role in regulating heat stress response in plants. Our previous study found that there were 82 non-redundant Hsfs in wheat, 18 of which belonged to subclass A2. In this study, we cloned an A2 member, TaHsfA2-1, which encoded a protein of 346 amino acid residues in wheat. The fusion protein TaHsfA2-1-GFP was localized in the nucleus under normal growth conditions. TaHsfA2-1 was expressed in nearly all the measured tissues, most highly in mature leaves. The expression level of TaHsfA2-1 can be enhanced by heat stress, PEG stress, and signal molecules such as H2O2 and SA. Yeast cells transformed with TaHsfA2-1 improved thermotolerance compared to those with the empty vector. TaHsfA2-1-overexpressing Arabidopsis displayed a better growth state with more green leaves than wild-type seedlings after heat stress. Accordingly, the chlorophyll content and survival rate in the transgenic lines were higher than in the wild type, and relative conductivity in the transgenic lines was lower than in the wild type. Further research found that TaHsfA2-1-overexpressing Arabidopsis up-regulated the expression of some heat shock protein genes (Hsps) compared to wild type after heat stress. These results suggested that TaHsfA2-1 is a new gene that improves thermotolerance in plants by mediating the expression of Hsps. A functional gene was provided for molecular breeding in the subsequent research.


Assuntos
Regulação da Expressão Gênica de Plantas/fisiologia , Fatores de Transcrição de Choque Térmico/genética , Proteínas de Plantas/genética , Termotolerância/genética , Triticum/fisiologia , Sequência de Aminoácidos , Arabidopsis/genética , Arabidopsis/fisiologia , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Fatores de Transcrição de Choque Térmico/química , Fatores de Transcrição de Choque Térmico/metabolismo , Filogenia , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Plântula/genética , Plântula/fisiologia , Alinhamento de Sequência , Triticum/genética
8.
Artigo em Inglês | MEDLINE | ID: mdl-32193092

RESUMO

Tomato (Lycopersicon esculentum Mill [Solanum lycopersicum L.].) is an important food material and cash crop, as well as a model plant for genetic evolution and molecular biology research. However, as a cold-sensitive crop originating from the tropics, the growth and development of tomato is often affected by low temperature stress. Therefore, how processing tomatoes resist this type of stress has important theoretical and practical significance. In this study, the LeCOLD1 gene was cloned from processing tomato. Subcellular localization analysis showed that LeCOLD1 was located in the plasma membrane. Real-time quantitative PCR analysis showed that LeCOLD1 was highly expressed in roots. Drought, salt and low temperatures induced the expression of COLD1. Overexpression and RNA interference vectors of LeCOLD1 were constructed and were transformed into tomato by the Agrobacterium-mediated method, and then obtaining transgenic tomato plants. It was found that LeCOLD1 increased the height of processing tomato plants and increased the length of their roots. In addition, overexpression of LeCOLD1 significantly improved the cold resistance of the plants. Overexpressing LeCOLD1 in tomato plants reduced the damage to the cell membrane, accumulation of ROS and photoinhibition of PSII, and maintained the high activity of antioxidant enzymes and the content of osmotic regulators. Further analysis revealed that during low temperature stress, the cells maintained high levels of antioxidant enzyme activity by regulating the transcription of the genes encoding these enzymes. The results show that overexpressing LeCOLD1 in tomato increases the plants' resistance to low temperatures, and that reducing LeCOLD1 expression makes the plants more sensitive to low temperatures.


Assuntos
Resposta ao Choque Frio , Lycopersicon esculentum/fisiologia , Temperatura Baixa , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Raízes de Plantas/genética , Plantas Geneticamente Modificadas/fisiologia
9.
Artigo em Inglês | MEDLINE | ID: mdl-32005383

RESUMO

In plants, the organ size is one of the most important features and regulated by an elaborate developmental program involving both internal and external signals. The steroidal hormone brassinosteroid (BR) plays an important role in regulating the organ size. BRASSINAZOLE RESISTANT 1 (BZR1) is one of important transcription factors that regulate organ size in BR signal pathway in Arabidopsis. The function of BZR1 on organ size is well characterized in Arabidopsis, but poorly understood in maize (Zea mays). To understand the mechanism of intrinsic organ size regulated by BZR1 during organogenesis, we identified the maize BZR1 and examined its function in Arabidopsis. Overexpression of ZmBZR1 displayed phenotypes of enlarged cotyledons, rosette leaves, floral organ and seed size in Arabidopsis. The cells in rosette leaves as well as other organs in transgenic ZmBZR1 lines were dramatically larger and longer than those in Col-0. ChIP and RNA-seq analysis showed ZmBZR1 can directly bind to the promoter region of organ size related genes, Germination Repression and Cell Expansion receptor-like kinase (GRACE) and KIP-RELATED PROTEIN6 (KRP6) to regulate their expression, suggesting ZmBZR1 is required for the progressive increase in cells during Arabidopsis development. Collectively, our findings provide significant insights into the mechanisms underlying regulation of organ size mediated by maize BZR1.


Assuntos
Organogênese Vegetal/genética , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Zea mays/fisiologia , Sequência de Aminoácidos , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/fisiologia , Proteínas de Ligação a DNA/química , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Plantas Geneticamente Modificadas/fisiologia , Sementes/fisiologia , Alinhamento de Sequência , Fatores de Transcrição/química , Fatores de Transcrição/metabolismo , Zea mays/genética , Zea mays/crescimento & desenvolvimento
10.
Artigo em Inglês | MEDLINE | ID: mdl-32005389

RESUMO

Salicylic acid (SA) plays an important role in the response of plants to abiotic stresses. Starvation stress affects plant cell metabolic activities, which further limits the normal growth and development of plants. It was reported that SA might play a regulatory role in the process of plant against starvation stress, but the mechanism involved in this process is still unclear. Thus, in this study, the transgenic plants overexpressing a SA binding protein 2 (SABP2) gene were exposed to starvation stress and the transgenic lines showed starvation-tolerant phenotype. Compared with wild-type (WT) plants, transgenic plants showed better growth status under poor-nutrition stress. Transgenic plants also showed more vigorous roots than WT plants. Physiological tests indicated that the transgenic plants showed higher relative water content (RWC), chlorophyll content, photosynthetic capacity, endogenous SA content, and lower ROS level compared to WT plants. Transcriptome analysis of tobacco plants identified 3, 748 differentially expressed genes (DEGs) between transgenic and WT plants under starvation stress. These DEGs are mainly involved in glycolysis/gluconeogenesis pathway group, MAPK signaling pathway group and plant hormone signal transduction pathway group. As determined by qPCR, up-regulated expression of fifteen genes such as abscisic acid receptor PYR1-like gene (NtPYR1-like), bidirectional sugar transporter N3-like gene (NtSWEETN3-like) and superoxide dismutase [Fe] chloroplastic-like gene (NtFeSOD-like), etc., was observed in transgenic plants under poor-nutrition stress which was in accordance with RNA-sequencing results. The modified pathways involved in plant hormone signaling are thought to be at least one of the main causes of the increased starvation tolerance of transgenic tobacco plants with altered SA homeostasis.


Assuntos
Esterases/genética , Regulação da Expressão Gênica de Plantas , Nutrientes/metabolismo , Proteínas de Plantas/genética , Ácido Salicílico/metabolismo , Tabaco/fisiologia , Esterases/metabolismo , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Estresse Fisiológico/genética , Tabaco/genética
11.
Artigo em Inglês | MEDLINE | ID: mdl-32062590

RESUMO

As a class of peptide hormone, plant natriuretic peptides (PNPs) play an important role in maintaining water and salt balance in plants, as well as in the physiological processes of biotic stress and pathogen resistance. However, in plants, except for some PNPs, such as the Arabidopsis thaliana PNP-A (AtPNP-A), of which the function has not yet been thoroughly revealed, few PNPs in other plants have been reported. In this study, a PNP-A (ScPNP-A) has been identified and characterized in Stellera chamaejasme for the first time. ScPNP-A is a double-psi beta-barrel (DPBB) fold containing protein and is localized in the extracellular (secreted) space. In S. chamaejasme, the expression of ScPNP-A was significantly up-regulated by salt, drought and cold stress. Changes at the physiological and biochemical levels and the expression of resistance-related genes indicated that overexpression of ScPNP-A can significantly improve salt, drought and freezing tolerance in Arabidopsis. ScPNP-A could stimulate the opening, not the closing of stomata, and its expression was not enhanced by external application of ABA. Furthermore, overexpression of ScPNP-A resulted in the elevated expression of genes in the ABA biosynthesis and reception pathway. These suggested that there may be some cross-talk between ScPNP-A and the ABA-dependent signaling pathways to regulate water related stress, however further experimentation is required to understand this relationship. In addition, overexpression of ScPNP-A can enhance the resistance to pathogens by enhancing SAR in Arabidopsis. These results indicate that ScPNP-A could function as a positive regulator in plant response to biotic stress and abiotic stress.


Assuntos
Arabidopsis , Peptídeos Natriuréticos , Estresse Fisiológico , Arabidopsis/genética , Arabidopsis/metabolismo , Secas , Expressão Gênica , Peptídeos Natriuréticos/genética , Peptídeos Natriuréticos/metabolismo , Estômatos de Plantas/genética , Estômatos de Plantas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Estresse Fisiológico/genética , Thymelaeaceae/genética
12.
Proc Natl Acad Sci U S A ; 117(6): 3281-3290, 2020 02 11.
Artigo em Inglês | MEDLINE | ID: mdl-31974310

RESUMO

There is considerable interest in engineering plant cell wall components, particularly lignin, to improve forage quality and biomass properties for processing to fuels and bioproducts. However, modifying lignin content and/or composition in transgenic plants through down-regulation of lignin biosynthetic enzymes can induce expression of defense response genes in the absence of biotic or abiotic stress. Arabidopsis thaliana lines with altered lignin through down-regulation of hydroxycinnamoyl CoA:shikimate/quinate hydroxycinnamoyl transferase (HCT) or loss of function of cinnamoyl CoA reductase 1 (CCR1) express a suite of pathogenesis-related (PR) protein genes. The plants also exhibit extensive cell wall remodeling associated with induction of multiple cell wall-degrading enzymes, a process which renders the corresponding biomass a substrate for growth of the cellulolytic thermophile Caldicellulosiruptor bescii lacking a functional pectinase gene cluster. The cell wall remodeling also results in the release of size- and charge-heterogeneous pectic oligosaccharide elicitors of PR gene expression. Genetic analysis shows that both in planta PR gene expression and release of elicitors are the result of ectopic expression in xylem of the gene ARABIDOPSIS DEHISCENCE ZONE POLYGALACTURONASE 1 (ADPG1), which is normally expressed during anther and silique dehiscence. These data highlight the importance of pectin in cell wall integrity and the value of lignin modification as a tool to interrogate the informational content of plant cell walls.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiologia , Lignina/metabolismo , Caules de Planta/metabolismo , Poligalacturonase/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Parede Celular/genética , Parede Celular/metabolismo , Pectinas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Poligalacturonase/genética
13.
Artigo em Inglês | MEDLINE | ID: mdl-31928668

RESUMO

Salinity threatens the productivity of tomato (Solanum lycopersicum L.). R2R3-type MYB transcription factors are important regulators in response to environmental stress. Here, we analyzed the function of the tomato R2R3-type MYB gene SlMYB102. A transcriptional activation assay showed that SlMYB102 had transactivation activity in yeast. Promoter analysis showed that multiple stress-related elements were found in the promoter of SlMYB102. Furthermore, SlMYB102 was induced by osmotic stress, particularly by salt stress. The overexpression of SlMYB102 in tomato affected multiple parameters under salinity stress. Under long-term salt stress, the degree of growth inhibition was significantly reduced in the two overexpression (OE) lines. In addition, the two OE lines maintained a better K+/Na+ ratio, lower reactive oxygen species (ROS) generation (O2•- production rate and H2O2 content) and lower electrolytic leakage rates than the wild type (WT). The activity of ROS scavenging enzymes including superoxide dismutase, peroxidase, catalase and ascorbate peroxidase, and the accumulation of antioxidants (ascorbic acid and glutathione) and proline was higher in the two OE lines compared with WT. The qRT-PCR analysis confirmed that the transcript abundance of many salt stress-related genes (SlSOS1, SlSOS2, SlNHX3, SlNHX4, SlHAK5, SlCPK1 and SlCPK3) was upregulated in two OE lines under salt stress. Collectively, these results suggest that SlMYB102 participates in tomato tolerance through the regulation of a series of molecular and physiological processes.


Assuntos
Regulação da Expressão Gênica de Plantas , Lycopersicon esculentum/fisiologia , Proteínas de Plantas/genética , Tolerância ao Sal/genética , Fatores de Transcrição/genética , Lycopersicon esculentum/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Salinidade , Fatores de Transcrição/metabolismo , Transcrição Genética
14.
Artigo em Inglês | MEDLINE | ID: mdl-31928677

RESUMO

NAC (NAM, ATAF1/2 and CUC2) proteins are plant-specific transcription factors (TFs) that are important in plant abiotic stress responses. In this study we isolated a NAC gene from Capsicum annuum leaves, designated as CaNAC064. We characterized the amino acid sequence of CaNAC064 and found that it contain conserved domains of the NAC family, including a highly conserved N-terminus domain and a highly variable C-terminus domain. Expression analysis showed that the 40C, 400C, salicylic acid (SA) and abscisic acid (ABA) treatments strongly induced the expression of CaNAC064 through silencing of CaNAC064 in pepper and overexpressing in Arabidopsis. CaNAC064-silenced pepper plants exhibited more serious wilting, higher MDA contents and chilling injury index, lower proline content, and more accumulation of ROS in the leaves after cold stress. The CaNAC064-overexpressing Arabidopsis plants exhibited lower MDA content, chilling injury index and relative electrolyte leakage content as compared to WT plants under cold stress. Transcriptional activation activity analysis indicated that CaNAC064 has transcriptional activation activity in the 691-1071 bp key region. We identified 45 proteins that putatively interact with CaNAC064 using the Yeast Two-Hybrid method. According to the Yeast Two-Hybrid and BIFC results, CaNAC064 interacted with low temperature-induced haplo-proteinase proteins in plant cell. These results suggested that CaNAC064 positively modulates plant cold-tolerance, laying the foundation for future investigations into the role of NACs as regulatory proteins of cold tolerance in plants.


Assuntos
Capsicum/fisiologia , Resposta ao Choque Frio/genética , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Sequência de Aminoácidos , Capsicum/genética , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Alinhamento de Sequência , Fatores de Transcrição/química , Fatores de Transcrição/metabolismo
15.
Int J Mol Sci ; 21(2)2020 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-31963715

RESUMO

Jatropha curcas L. is monoecious with a low female-to-male ratio, which is one of the factors restricting its seed yield. Because the phytohormone cytokinins play an essential role in flower development, particularly pistil development, in this study, we elevated the cytokinin levels in J. curcas flowers through transgenic expression of a cytokinin biosynthetic gene (AtIPT4) from Arabidopsis under the control of a J. curcas orthologue of TOMATO MADS BOX GENE 6 (JcTM6) promoter that is predominantly active in flowers. As expected, the levels of six cytokinin species in the inflorescences were elevated, and flower development was modified without any alterations in vegetative growth. In the transgenic J. curcas plants, the flower number per inflorescence was significantly increased, and most flowers were pistil-predominantly bisexual, i.e., the flowers had a huge pistil surrounded with small stamens. Unfortunately, both the male and the bisexual flowers of transgenic J. curcas were infertile, which might have resulted from the continuously high expression of the transgene during flower development. However, the number and position of floral organs in the transgenic flowers were well defined, which suggested that the determinacy of the floral meristem was not affected. These results suggest that fine-tuning the endogenous cytokinins can increase the flower number and the female-to-male ratio in J. curcas.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/metabolismo , Citocininas/metabolismo , Jatropha/genética , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Vias Biossintéticas , Citocininas/genética , Flores/genética , Flores/fisiologia , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Inflorescência , Jatropha/fisiologia , Plantas Geneticamente Modificadas/fisiologia , Regiões Promotoras Genéticas , Reprodução Assexuada
16.
Plant Cell Rep ; 39(4): 511-525, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31938834

RESUMO

KEY MESSAGE: A simple and robust Agrobacterium-mediated gene expression system in the C4 panicoid model crop, foxtail millet has been developed with up to 27 % transformation efficiency. Foxtail millet (Setaria italica L.) is a model crop to study C4 photosynthesis, abiotic stress tolerance, and bioenergy traits. Advances in molecular genetics and genomics had identified several potential genes in this crop that would serve as candidates for imparting climate-resilient traits in related millets, cereals, and biofuel crops. However, the lack of an efficient genetic transformation system has been impeding the functional characterization of these genes in foxtail millet per se. Given this, an easy and efficient regeneration and transformation protocol was optimized using mature seeds as a choicest explant. The suitability of secondary embryogenic calli over primary calli is underlined due to their high competence. The use of perfect combinations of plant growth regulators together with the ionic strength of organic and inorganics salts was found to influence regeneration and genetic transformation. We studied and optimized various crucial factors that affect the genetic transformation of foxtail millet calli using Agrobacterium tumefaciens-mediated approach. Secondary embryogenic calli and LBA44404 strain were found to be the best targets for transformation. The use of high sucrose and glucose, together with freshly prepared tobacco leaves extract, Silwet L-77 and acetosyringone, improved the efficiency of the genetic transformation of foxtail millet. Moreover, the use of an in vitro regeneration system with 84% callusing efficiency and 70-74% regeneration frequency led to a high recovery of transformants. Altogether, the present study reports a highly efficient (~ 27%) transformation system in foxtail millet that will expedite forward and reverse genetic studies in this important crop.


Assuntos
Agrobacterium tumefaciens/genética , Produtos Agrícolas/genética , Setaria (Planta)/genética , Transformação Genética , Técnicas de Cultura de Células/métodos , Células Cultivadas , Técnicas Genéticas , Vetores Genéticos , Fenótipo , Células Vegetais/efeitos dos fármacos , Células Vegetais/microbiologia , Células Vegetais/fisiologia , Reguladores de Crescimento de Planta/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Regeneração/genética , Regeneração/fisiologia , Sementes/efeitos dos fármacos , Sementes/genética , Sementes/metabolismo , Sementes/microbiologia , Setaria (Planta)/metabolismo , Setaria (Planta)/microbiologia
17.
Planta ; 251(2): 38, 2020 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-31907623

RESUMO

MAIN CONCLUSION: miRNAs control leaf size of maize crop during chilling stress tolerance by regulating developmentally important transcriptional factors and sustaining redox homeostasis of cells. Chilling temperature (0-15 °C) is a major constraint for the cultivation of maize (Zea mays) which inhibits the early growth of maize leading to reduction in leaf size. Growth and development take place in meristem, elongation, and mature zones that are linearly located along the leaf base to tip. To prevent shortening of leaf caused by chilling, this study aims to elucidate the regulatory roles of microRNA (miRNA) genes in the controlling process switching between growth and developmental stages. In this respect, hybrid maize ADA313 seedlings were treated to the chilling temperature which caused 26% and 29% reduction in the final leaf length and a decline in cell production of the fourth leaf. The flow cytometry data integrated with the expression analysis of cell cycle genes indicated that the reason for the decline was a failure proceeding from G2/M rather than G1/S. Through an miRNome analysis of 321 known maize miRNAs, 24, 6, and 20 miRNAs were assigned to putative meristem, elongation, and mature zones, respectively according to their chilling response. To gain deeper insight into decreased cell production, in silico, target prediction analysis was performed for meristem specific miRNAs. Among the miRNAs, miR160, miR319, miR395, miR396, miR408, miR528, and miR1432 were selected for confirming the potential of negative regulation with their predicted targets by qRT-PCR. These findings indicated evidence for improvement of growth and yield under chilling stress of the maize.


Assuntos
MicroRNAs/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , RNA de Plantas/metabolismo , Zea mays/metabolismo , Zea mays/fisiologia , Temperatura Baixa , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , MicroRNAs/genética , Folhas de Planta/genética , Folhas de Planta/metabolismo , Folhas de Planta/fisiologia , RNA de Plantas/genética , Zea mays/genética
18.
Plant Mol Biol ; 102(4-5): 447-462, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31898148

RESUMO

KEY MESSAGE: ZjICE2 works as a positive regulator in abiotic stress responses and ZjICE2 is a valuable genetic resource to improve abiotic stress tolerance in the molecular breeding program of Zoysia japonica. The basic helix-loop-helix (bHLH) family transcription factors (TFs) play an important role in response to biotic or abiotic stresses in plants. However, the functions of bHLH TFs in Zoysia japonica, one of the warm-season turfgrasses, remain poorly understood. Here, we identified ZjICE2 from Z. japonica, a novel MYC-type bHLH transcription factor that was closely related to ICE homologs in the phylogenetic tree, and its expression was regulated by various abiotic stresses. Transient expression of ZjICE2-GFP in onion epidermal cells revealed that ZjICE2 was a nuclear-localized protein. Also, ZjICE2 bound the MYC cis-element in the promoter of dehydration responsive element binding 1 of Z. japonica (ZjDREB1) using yeast one-hybrid assay. A phenotypic analysis showed that overexpression of the ZjICE2 in Arabidopsis enhanced tolerance to cold, drought, and salt stresses. The transgenic Arabidopsis and Z. japonica accumulated more transcripts of cold-responsive DREB/CBFs and their downstream genes than the wild type (WT) after cold treatment. Furthermore, the transgenic plants exhibited an enhanced Reactive oxygen species (ROS) scavenging ability, which resulted in an efficient maintenance of oxidant-antioxidant homeostasis. In addition, overexpression of the ZjICE2 in Z. japonica displayed intensive cold tolerance with increases in chlorophyll contents and photosynthetic efficiency. Our study suggests that ZjICE2 works as a positive regulator in abiotic stress responses and the ICE-DREB/CBFs response pathway involved in cold stress tolerance is also conserved in Z. japonica. These results provide a valuable genetic resource for the molecular breeding program especially for warm-season grasses as well as other leaf crop plants.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/fisiologia , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/fisiologia , Poaceae/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Estresse Fisiológico , Arabidopsis/genética , Arabidopsis/fisiologia , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/fisiologia , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Temperatura Baixa , Resposta ao Choque Frio , Secas , Filogenia , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/fisiologia , Poaceae/genética , Regulon , Tolerância ao Sal , Fatores de Transcrição/genética , Fatores de Transcrição/fisiologia , Ativação Transcricional
19.
Cell Mol Life Sci ; 77(4): 705-718, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31250033

RESUMO

Oxidative stress can lead to plant growth retardation, yield loss, and death. The atr7 mutant of Arabidopsis thaliana exhibits pronounced tolerance to oxidative stress. Using positional cloning, confirmed by knockout and RNA interference (RNAi) lines, we identified the atr7 mutation and revealed that ATR7 is a previously uncharacterized gene with orthologs in other seed plants but with no homology to genes in lower plants, fungi or animals. Expression of ATR7-GFP fusion shows that ATR7 is a nuclear-localized protein. RNA-seq analysis reveals that transcript levels of genes encoding abiotic- and oxidative stress-related transcription factors (DREB19, HSFA2, ZAT10), chromatin remodelers (CHR34), and unknown or uncharacterized proteins (AT5G59390, AT1G30170, AT1G21520) are elevated in atr7. This indicates that atr7 is primed for an upcoming oxidative stress via pathways involving genes of unknown functions. Collectively, the data reveal ATR7 as a novel seed plants-specific nuclear regulator of oxidative stress response.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Sementes/genética , Arabidopsis/fisiologia , Genes de Plantas , Mutação , Estresse Oxidativo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Fatores de Transcrição/genética
20.
Physiol Plant ; 168(1): 38-57, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30714160

RESUMO

In plants, CALCIUM-DEPENDENT PROTEIN KINASES (CDPKs/CPKs) are involved in calcium signaling in response to endogenous and environmental stimuli. Here, we report that ZmCPK11, one of maize CDPKs, participates in salt stress response and tolerance. Salt stress induced expression and upregulated the activity of ZmCPK11 in maize roots and leaves. Activation of ZmCPK11 upon salt stress was also observed in roots and leaves of transgenic Arabidopsis plants expressing ZmCPK11. The transgenic plants showed a long-root phenotype under control conditions and a short-root phenotype under NaCl, abscisic acid (ABA) or jasmonic acid (JA) treatment. Analysis of ABA and JA content in roots indicated that ZmCPK11 can mediate root growth by regulating the levels of these phytohormones. Moreover, 4-week-old transgenic plants were more tolerant to salinity than the wild-type plants. Their leaves were less chlorotic and showed weaker symptoms of senescence accompanied by higher chlorophyll content and higher quantum efficiency of photosystem II. The expression of Na+ /K+ transporters (HKT1, SOS1 and NHX1) and transcription factors (CBF1, CBF2, CBF3, ZAT6 and ZAT10) with known links to salinity tolerance was upregulated in roots of the transgenic plants upon salt stress. Furthermore, the transgenic plants accumulated less Na+ in roots and leaves under salinity, and showed a higher K+ /Na+ ratio in leaves. These results show that the improved salt tolerance in ZmCPK11-transgenic plants could be due to an upregulation of genes involved in the maintenance of intracellular Na+ and K+ homeostasis and a protection of photosystem II against damage.


Assuntos
Arabidopsis/fisiologia , Complexo de Proteína do Fotossistema II/metabolismo , Proteínas Quinases/metabolismo , Tolerância ao Sal , Zea mays/enzimologia , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Homeostase , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas , Plantas Geneticamente Modificadas/fisiologia , Potássio/análise , Proteínas Quinases/genética , Sódio , Simportadores de Cloreto de Sódio-Potássio , Fatores de Transcrição , Zea mays/genética
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