Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 4.366
Filtrar
1.
Plant Sci ; 290: 110284, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31779918

RESUMO

The first step in the Phosphorylated Pathway of serine (Ser) Biosynthesis (PPSB) is catalyzed by the enzyme Phosphoglycerate Dehydrogenase (PGDH), coded in Arabidopsis thaliana by three genes. Gene expression analysis indicated that PGDH1 and PGDH2 were induced, while PGDH3 was repressed, by salt-stress. Accordingly, PGDH3 overexpressing plants (Oex PGDH3) were more sensitive to salinity than wild type plants (WT), while plants overexpressing PGDH1 (Oex PGDH1) performed better than WT under salinity conditions. Oex PGDH1 lines displayed lower levels of the salt-stress markers proline and raffinose in roots than WT under salt-stress conditions. Besides, the ratio of oxidized glutathione (GSSG) without and with salt-stress was the highest in Oex PGDH1, and the lowest in Oex PGDH3 compared to WT. These results corroborated that PGDH3 activity could be detrimental, while PGDH1 activity could be beneficial for plant salt tolerance. Under salt-stress conditions, PGDH1 overexpression increased Ser content only in roots, while PGDH3 overexpression increased the amino acid level in both aerial parts and roots, compared to the WT. Our results indicate that the response of PGDH family genes to salt-stress depends on the specific gene studied and that increases in Ser content are not always correlated with enhanced plant salt tolerance.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/fisiologia , Regulação da Expressão Gênica de Plantas/fisiologia , Família Multigênica/fisiologia , Fosfoglicerato Desidrogenase/genética , Tolerância ao Sal/genética , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Fosfoglicerato Desidrogenase/metabolismo , Raízes de Plantas/metabolismo
2.
Plant Mol Biol ; 102(3): 287-306, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31872308

RESUMO

KEY MESSAGE: At the early stage of pollination, the difference in gene expression between compatibility and incompatibility is highly significant about the pollen-specific expression of the LRR gene, resistance, and defensin genes. In Rosaceae, incompatible pollen can penetrate into the style during the gametophytic self-incompatibility response. It is therefore considered a stylar event rather than a stigmatic event. In this study, we explored the differences in gene expression between compatibility and incompatibility in the early stage of pollination. The self-compatible pear variety "Jinzhuili" is a naturally occurring bud mutant from "Yali", a leading Chinese native cultivar exhibiting typical gametophytic self-incompatibility. We collected the styles of 'Yali' and 'Jinzhuili' at 0.5 and 2 h after self-pollination and then performed high-throughput sequencing. According to the KEGG analysis of the differentially expressed genes, several metabolic pathways, such as "Plant hormone signal transduction", "Plant-pathogen interaction", are the main pathways was the most represented pathway. Quantitative PCR was used to validate these differential genes. The expression levels of genes related to pollen growth and disease inhibition, such as LRR (Leucine-rich repeat extensin), resistance, defensin, and auxin, differed significantly between compatible and incompatible pollination. Interestingly, at 0.5 h, most of these genes were upregulated in the compatible pollination system compared with the incompatible pollination system. Calcium transport, which requires ATPase, also demonstrated upregulated expression. In summary, the self-incompatibility reaction was initiated when the pollen land on the stigma.


Assuntos
Pólen/genética , Polinização/genética , Polinização/fisiologia , Pyrus/genética , Pyrus/fisiologia , /métodos , Morte Celular , Técnicas de Reprogramação Celular , Regulação da Expressão Gênica de Plantas/fisiologia , Genes de Plantas/genética , Ácidos Indolacéticos , Oxigenases/genética , Reguladores de Crescimento de Planta , Proteínas de Plantas/genética , Pólen/crescimento & desenvolvimento
3.
PLoS Pathog ; 15(12): e1008110, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31790500

RESUMO

Viroids are small, non-protein-coding RNAs which can induce disease symptoms in a variety of plant species. Potato (Solanum tuberosum L.) is the natural host of Potato spindle tuber viroid (PSTVd) where infection results in stunting, distortion of leaves and tubers and yield loss. Replication of PSTVd is accompanied by the accumulation of viroid-derived small RNAs (sRNAs) proposed to play a central role in disease symptom development. Here we report that PSTVd sRNAs direct RNA silencing in potato against StTCP23, a member of the TCP (teosinte branched1/Cycloidea/Proliferating cell factor) transcription factor family genes that play an important role in plant growth and development as well as hormonal regulation, especially in responses to gibberellic acid (GA). The StTCP23 transcript has 21-nucleotide sequence complementarity in its 3' untranslated region with the virulence-modulating region (VMR) of PSTVd strain RG1, and was downregulated in PSTVd-infected potato plants. Analysis using 3' RNA ligase-mediated rapid amplification of cDNA ends (3' RLM RACE) confirmed cleavage of StTCP23 transcript at the expected sites within the complementarity with VMR-derived sRNAs. Expression of these VMR sRNA sequences as artificial miRNAs (amiRNAs) in transgenic potato plants resulted in phenotypes reminiscent of PSTVd-RG1-infected plants. Furthermore, the severity of the phenotypes displayed was correlated with the level of amiRNA accumulation and the degree of amiRNA-directed down-regulation of StTCP23. In addition, virus-induced gene silencing (VIGS) of StTCP23 in potato also resulted in PSTVd-like phenotypes. Consistent with the function of TCP family genes, amiRNA lines in which StTCP23 expression was silenced showed a decrease in GA levels as well as alterations to the expression of GA biosynthesis and signaling genes previously implicated in tuber development. Application of GA to the amiRNA plants minimized the PSTVd-like phenotypes. Taken together, our results indicate that sRNAs derived from the VMR of PSTVd-RG1 direct silencing of StTCP23 expression, thereby disrupting the signaling pathways regulating GA metabolism and leading to plant stunting and formation of small and spindle-shaped tubers.


Assuntos
Genes de Plantas , Doenças das Plantas/virologia , Solanum tuberosum/virologia , Viroides/patogenicidade , Virulência/fisiologia , Regulação da Expressão Gênica de Plantas/fisiologia , Interferência de RNA/fisiologia , Vírus de RNA , RNA Viral , Solanum tuberosum/genética , Fatores de Transcrição
4.
BMC Plant Biol ; 19(1): 465, 2019 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-31684878

RESUMO

BACKGROUND: Fruit coloration is one of the main quality parameters of Citrus fruit primarily determined by genetic factors. The fruit of ordinary sweet orange (Citrus sinensis) displays a pleasant orange tint due to accumulation of carotenoids, representing ß,ß-xanthophylls more than 80% of the total content. 'Pinalate' is a spontaneous bud mutant, or somatic mutation, derived from sweet orange 'Navelate', characterized by yellow fruits due to elevated proportions of upstream carotenes and reduced ß,ß-xanthophylls, which suggests a biosynthetic blockage at early steps of the carotenoid pathway. RESULTS: To identify the molecular basis of 'Pinalate' yellow fruit, a complete characterization of carotenoids profile together with transcriptional changes in carotenoid biosynthetic genes were performed in mutant and parental fruits during development and ripening. 'Pinalate' fruit showed a distinctive carotenoid profile at all ripening stages, accumulating phytoene, phytofluene and unusual proportions of 9,15,9'-tri-cis- and 9,9'-di-cis-ζ-carotene, while content of downstream carotenoids was significantly decreased. Transcript levels for most of the carotenoid biosynthetic genes showed no alterations in 'Pinalate'; however, the steady-state level mRNA of ζ-carotene isomerase (Z-ISO), which catalyses the conversion of 9,15,9'-tri-cis- to 9,9'-di-cis-ζ-carotene, was significantly reduced both in 'Pinalate' fruit and leaf tissues. Isolation of the 'Pinalate' Z-ISO genomic sequence identified a new allele with a single nucleotide insertion at the second exon, which generates an alternative splicing site that alters Z-ISO transcripts encoding non-functional enzyme. Moreover, functional assays of citrus Z-ISO in E.coli showed that light is able to enhance a non-enzymatic isomerization of tri-cis to di-cis-ζ-carotene, which is in agreement with the partial rescue of mutant phenotype when 'Pinalate' fruits are highly exposed to light during ripening. CONCLUSION: A single nucleotide insertion has been identified in 'Pinalate' Z-ISO gene that results in truncated proteins. This causes a bottleneck in the carotenoid pathway with an unbalanced content of carotenes upstream to ß,ß-xanthophylls in fruit tissues. In chloroplastic tissues, the effects of Z-ISO alteration are mainly manifested as a reduction in total carotenoid content. Taken together, our results indicate that the spontaneous single nucleotide insertion in Z-ISO is the molecular basis of the yellow pigmentation in 'Pinalate' sweet orange and points this isomerase as an essential activity for carotenogenesis in citrus fruits.


Assuntos
Citrus sinensis/fisiologia , Frutas/fisiologia , Regulação da Expressão Gênica de Plantas/fisiologia , Isomerases/genética , Proteínas de Plantas/genética , Alelos , Sequência de Aminoácidos , Citrus sinensis/genética , Cor , Frutas/genética , Isomerases/química , Isomerases/metabolismo , Pigmentação/genética , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Alinhamento de Sequência
5.
Plant Mol Biol ; 101(6): 537-550, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31745746

RESUMO

KEY MESSAGE: MIR159/319 have conserved evolution and diversified function after WGT in Brassica campestris, both of them can lead pollen vitality and germination abnormality, Bra-MIR319c also can function in flower development. MiR159 and miR319 are extensively studied highly conserved microRNAs which play roles in vegetative development, reproduction, and hormone regulation. In this study, the effects of whole-genome triplication (WGT) on the evolution of the MIR159/319 family and the functional diversification of the genes were comprehensively investigated in Brassica campestris. We identified 11 MIR159/319 genes in B. campestris, which produced five mature sequences. After analyzing the precursor sequences and phylogenetic tree, we found that Bra-MIR159/319 have evolutionary conservatism. Furthermore, Bra-MIR159/319 show functional diversification after WGT, as indicated by their expression patterns and the cis-element in their promoter. GUS signal showed that Bra-MIR159a and Bra-MIR319c can be expressed in anther but in different development stages. In B. campestris, overexpressed MIR159a and MIR319c contribute to late anther development and promote pollen abortion. Moreover, Bra-MIR319c can partially assume the function of MIR319a in flower development.


Assuntos
Brassica/metabolismo , Proteínas de Plantas/metabolismo , Pólen/metabolismo , Brassica/genética , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Proteínas de Plantas/genética , Pólen/genética
6.
Plant Physiol Biochem ; 144: 455-465, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31655344

RESUMO

Sugarcane is an important sugar and energy crop worldwide. It utilises highly efficient C4 photosynthesis and accumulates sucrose in its culms. The sucrose content in sugarcane culms is a quantitative trait controlled by multiple genes. The regulatory mechanism underlying the maximum sucrose level in sugarcane culms remains unclear. We used transcriptome sequences to identify the potential regulatory genes involved in sucrose accumulation in Saccarum officinarum L. cv. Badila. The sucrose accumulating internodes at the elongation and mature growth stage and the immature internodes with low sucrose content at the mature stage were used for RNA sequencing. The obtained differentially expressed genes (DEGs) related to sucrose accumulation were analysed. Results showed that the transcripts encoding invertase (beta-fructofuranosidase, EC: 3.2.1.26) which catalyses sucrose hydrolysis and 6-phosphofructokinase (PFK, EC: 2.7.1.11), a key glycolysis regulatory enzyme, were downregulated in the high sucrose accumulation internodes. The transcripts encoding key enzymes for ABA, gibberellin and ethylene synthesis were also downregulated during sucrose accumulation. Furthermore, regulated protein kinase, transcription factor and sugar transporter genes were also obtained. This research can clarify the molecular regulation network of sucrose accumulation in sugarcane.


Assuntos
Saccharum/metabolismo , Sacarose/metabolismo , Transcriptoma/genética , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Saccharum/genética
7.
BMC Plant Biol ; 19(1): 423, 2019 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-31610785

RESUMO

BACKGROUND: Pink-flowered strawberry is a promising new ornamental flower derived from intergeneric hybridization (Fragaria × Potentilla) with bright color, a prolonged flowering period and edible fruits. Its flower color ranges from light pink to red. Pigment compounds accumulated in its fruits were the same as in cultivated strawberry fruits, but different from that in its flowers. However, the transcriptional events underlying the anthocyanin biosynthetic pathway have not been fully characterized in petal coloration. To gain insights into the regulatory networks related to anthocyanin biosynthesis and identify the key genes, we performed an integrated analysis of the transcriptome and metabolome in petals of pink-flowered strawberry. RESULTS: The main pigments of red and dark pink petals were anthocyanins, among which cyanidins were the main compound. There were no anthocyanins detected in the white-flowered hybrids. A total of 50,285 non-redundant unigenes were obtained from the transcriptome databases involved in red petals of pink-flowered strawberry cultivar Sijihong at three development stages. Amongst the unigenes found to show significant differential expression, 57 were associated with anthocyanin or other flavonoid biosynthesis, in which they were regulated by 241 differentially expressed members of transcription factor families, such as 40 MYBs, 47 bHLHs, and 41 NACs. Based on a comprehensive analysis relating pigment compounds to gene expression profiles, the mechanism of flower coloration was examined in pink-flowered strawberry. A new hypothesis was proposed to explain the lack of color phenotype of the white-flowered strawberry hybrids based on the transcriptome analysis. The expression patterns of FpDFR and FpANS genes corresponded to the accumulation patterns of cyanidin contents in pink-flowered strawberry hybrids with different shades of pink. Moreover, FpANS, FpBZ1 and FpUGT75C1 genes were the major factors that led to the absence of anthocyanins in the white petals of pink-flowered strawberry hybrids. Meanwhile, the competitive effect of FpFLS and FpDFR genes might further inhibit anthocyanin synthesis. CONCLUSIONS: The data presented herein are important for understanding the molecular mechanisms underlying the petal pigmentation and will be powerful for integrating novel potential target genes to breed valuable pink-flowered strawberry cultivars.


Assuntos
Antocianinas/metabolismo , Flores/metabolismo , Fragaria/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Transcriptoma , Cor , Fragaria/metabolismo
8.
BMC Plant Biol ; 19(1): 419, 2019 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-31604421

RESUMO

BACKGROUND: Eruca vesicaria subsp. sativa is one of the Cruciferae species most tolerant to drought stress. In our previous study some extremely drought-tolerant/sensitive Eruca lines were obtained. However little is known about the mechanism for drought tolerance in Eruca. METHODS: In this study two E. vesicaria subs. sativa lines with contrasting drought tolerance were treated with liquid MS/PEG solution. Total RNA was isolated from 7-day old whole seedlings and then applied to Illumina sequencing platform for high-throughput transcriptional sequencing. RESULTS: KEGG pathway analysis indicated that differentially expressed genes (DEGs) involved in alpha-Linolenic acid metabolism, Tyrosine metabolism, Phenylalanine, Tyrosine and tryptophan biosynthesis, Galactose metabolism, Isoquinoline alkaloid biosynthesis, Tropane, Piperidine and pyridine alkaloid biosynthesis, Mineral absorption, were all up-regulated specifically in drought-tolerant (DT) Eruca line under drought stress, while DEGs involved in ribosome, ribosome biogenesis, Pyrimidine metabolism, RNA degradation, Glyoxylate and dicarboxylate metabolism, Aminoacyl-tRNA biosynthesis, Citrate cycle, Methane metabolism, Carbon fixation in photosynthetic organisms, were all down-regulated. 51 DEGs were found to be most significantly up-regulated (log2 ratio ≥ 8) specifically in the DT line under PEG treatment, including those for ethylene-responsive transcription factors, WRKY and bHLH transcription factors, calmodulin-binding transcription activator, cysteine-rich receptor-like protein kinase, mitogen-activated protein kinase kinase, WD repeat-containing protein, OPDA reductase, allene oxide cyclase, aquaporin, O-acyltransferase WSD1, C-5 sterol desaturase, sugar transporter ERD6-like 12, trehalose-phosphate phosphatase and galactinol synthase 4. Eight of these 51 DEGs wre enriched in 8 COG and 17 KEGG pathways. CONCLUSIONS: DEGs that were found to be most significantly up-regulated specifically in the DT line under PEG treatment, up-regulation of DEGs involved in Arginine and proline metabolism, alpha-linolenic acid metabolism and down-regulation of carbon fixation and protein synthesis might be critical for the drought tolerance in Eruca. These results will be valuable for revealing mechanism of drought tolerance in Eruca and also for genetic engineering to improve drought tolerance in crops.


Assuntos
Brassicaceae/fisiologia , Regulação da Expressão Gênica de Plantas/fisiologia , Polietilenoglicóis/administração & dosagem , Estresse Fisiológico/fisiologia , Transcriptoma/fisiologia , Brassicaceae/genética , Secas , Perfilação da Expressão Gênica , Sequenciamento de Nucleotídeos em Larga Escala , Estresse Fisiológico/genética
9.
Plant Physiol Biochem ; 144: 312-323, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31606716

RESUMO

Plants are subjected to a variety of abiotic stresses during their lifetime, and drought and salt stress are some of the main causes of reduced crop yields. Previous studies have shown that AREB/ABFs within bZIP transcription factors are involved in plant drought and salt stress responses in an ABA-dependent manner. However, the properties and functions of AREB/ABFs in Fagopyrum tataricum, a cereal with good resistance to abiotic stresses, are poorly understood. In this study, a gene encoding an AREB/ABF, designated FtbZIP83, was first isolated from Tartary buckwheat. Expression analysis in Tartary buckwheat indicated that FtbZIP83 was significantly induced by abscisic acid (ABA), NaCl and polyethylene glycol (PEG). The overexpression of FtbZIP83 in Arabidopsis resulted in increased drought/salt tolerance, which was attributed not only to higher proline (Pro) contents and antioxidant enzyme activity in transgenic lines compared with controls but also to the lower reactive oxygen species (ROS) accumulation and malondialdehyde (MDA) content. In addition, we found that FtbZIP83 was able to respond to drought and salt stress by upregulating the transcript abundance of downstream ABA-inducible gene. Furthermore, promoter sequence analysis showed that ABREs were present, and the activity of the FtbZIP83 promoter in transgenic Arabidopsis after drought stress was significantly higher than that under normal conditions. Based on the potential signalling pathways involved in AREB/ABFs, we also screened for the interaction protein FtSnRK2.6/2.3, which may phosphorylate FtbZIP83. Collectively, these results provide evidence that FtbZIP83, as a positive regulator, responds to drought/salt stress via an ABA-dependent signalling pathway composed of SnRK2-AREB/ABF.


Assuntos
Secas , Fagopyrum/metabolismo , Fatores de Transcrição/metabolismo , Ácido Abscísico/metabolismo , Fagopyrum/genética , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/efeitos dos fármacos , Plantas Geneticamente Modificadas/genética , Tolerância ao Sal/genética , Tolerância ao Sal/fisiologia , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Fatores de Transcrição/genética
10.
Plant Sci ; 289: 110215, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31623776

RESUMO

14-3-3 proteins are a family of conserved proteins present in eukaryotes as several isoforms, playing a regulatory role in many cellular and physiological processes. In plants, 14-3-3 proteins have been reported to be involved in the response to stress conditions, such as drought, salt and cold. In the present study, 14-3-3ε and 14-3-3ω isoforms, which were representative of ε and non-ε phylogenetic groups, were overexpressed in Arabidopsis thaliana plants; the effect of their overexpression was investigated on H+-ATPase activation and plant response to cold stress. Results demonstrated that H+-ATPase activity was increased in 14-3-3ω-overexpressing plants, whereas overexpression of both 14-3-3 isoforms brought about cold stress tolerance, which was evaluated through ion leakage, lipid peroxidation, osmolyte synthesis, and ROS production assays. A dedicated tandem mass tag (TMT)-based proteomic analysis demonstrated that different proteins involved in the plant response to cold or oxidative stress were over-represented in 14-3-3ε-overexpressing plants.


Assuntos
Proteínas 14-3-3/genética , Arabidopsis/genética , Temperatura Baixa , Regulação da Expressão Gênica de Plantas/fisiologia , Proteínas de Plantas/genética , Estresse Fisiológico/genética , Proteínas 14-3-3/metabolismo , Aclimatação/genética , Arabidopsis/enzimologia , Arabidopsis/metabolismo , Proteínas de Plantas/metabolismo , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , ATPases Vacuolares Próton-Translocadoras/genética , ATPases Vacuolares Próton-Translocadoras/metabolismo
11.
Plant Physiol Biochem ; 144: 334-344, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31622936

RESUMO

Constitutive overexpression of the rice heterotrimeric G protein beta subunit gene (RGB1) in the commercial rice cultivar BRRI Dhan 55 resulted in improved tolerance to heat or salinity or their combination. Two independently in planta transformed plants with the gene confirmed to be integrated at T2 by Southern hybridization and showing high expression at the T3 seedling stage showed better physiological performance after 8 days in 120 mM salt stress than the wild type. The plants had significantly lower electrolyte leakage and malondialdehyde production, while showing higher levels of chlorophyll. Significantly higher germination at 48 °C or with combined stresses of 42/40 °C day/night stress in the presence of 120 mM salt for 2 days was also observed. Stress responsive genes such as OsAPX1, OsSOD, OsHKT1, OsHSP1, OsHSP2 and OsCOR47 showed higher expression in the RGB1 positive plants. These RGB1 transgenic plants can likely provide a strong defense against climate change.


Assuntos
Oryza/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Temperatura Alta , Oryza/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Tolerância ao Sal/genética , Tolerância ao Sal/fisiologia
12.
Plant Physiol Biochem ; 144: 375-385, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31622940

RESUMO

Soybeans are known for its good source of protein (40%), oil (20%) and also serve as a source of nutraceutical compounds including tocopherols (toc). To know the molecular basis of differential α-toc accumulation in two contrasting soybean genotypes: DS74 (low α-toc - 1.36 µg/g and total-toc -29.72 µg/g) and Bragg (high α-toc - 10.48 µg/g and total-toc 178.91 µg/g), the analysis of γ-TMT3 promoter activity and its methylation patterns were carried out. The sequencing results revealed nucleotide variation between Bragg:γ-TMT3-P and DS74:γ-TMT3-P, however none of the variations were found in core-promoter region or in cis-elements. The histochemical GUS assay revealed higher promoter activity of Bragg:γ-TMT3-P than that of DS74:γ-TMT3-P and correlated with significantly higher and lower (P < 0.05) expression of γ-TMT3 gene respectively. To know the molecular basis of differential accumulation of α-toc in these contrasting soybean genotypes, the DNA methylation pattern of γ-TMT3 gene body and its promoter was studied in both varieties. The results showed higher percentage (62.5%) of methylation in DS74:γ-TMT3-P than in Bragg:γ-TMT3-P (50%). Out of all the methylation sites in the promoter region, one of methylation site was found at CAAT box (-190 bp) of DS74:γ-TMT3-P. Further gene body methylation patterns revealed lowest % (40%) of CG methylation in DS74:γ-TMT3 gene as compared to Bragg:γ-TMT3 (64.2%). Thus our study revealed that, expression of γ-TMT3 gene was influenced by its promoter activity and methylation patterns in cis-elements of γ-TMT3 promoter and gene body. This study will help us to understand the possible role of methylation and promoter activity in determining the α-toc content in soybean seeds.


Assuntos
Soja/metabolismo , Tocoferóis/metabolismo , alfa-Tocoferol/metabolismo , gama-Tocoferol/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Regiões Promotoras Genéticas/genética
13.
Plant Physiol Biochem ; 143: 190-202, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31518850

RESUMO

Temperature is one of the most important environmental factors limiting tea plant growth and tea production. Previously we reported that both Ca2+ and ROS signals play important roles in tea plant cold acclimation. Here, we identified 26 CsCPK transcripts, analyzed their phylogenetic and sequence characters, and detected their transcriptions to monitor Ca2+ signaling status. Tissue-specific expression profiles indicated that most CsCPK genes were constitutively expressed in tested tissues, suggesting their possible roles in development. Cold along with calcium inhibitor assays suggested that CsCPKs are important cold regulators and CsCPK30/5/4/9 maybe the key members. Moreover, LaCl3 or EGTA pre-treatment could result in impaired Ca2+ signaling and compromised cold-responding network, but higher catechins accumulation revealed their potential positive roles in cold responses. Those findings indicated that catechins and other secondary metabolites in tea plant may form an alternative cold-responding network that closely correlated with Ca2+ signaling status.


Assuntos
Camellia sinensis/metabolismo , Catequina/metabolismo , Proteínas Quinases/metabolismo , Camellia sinensis/genética , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteínas Quinases/genética
14.
Plant Cell Physiol ; 60(11): 2360-2368, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31529098

RESUMO

The circadian clock is a timekeeping system for regulation of numerous biological daily rhythms. One characteristic of the circadian clock is that period length remains relatively constant in spite of environmental fluctuations, such as temperature change. Here, using the curated collection of in-house small molecule chemical library (ITbM chemical library), we show that small molecule 3,4-dibromo-7-azaindole (B-AZ) lengthened the circadian period of Arabidopsis thaliana (Arabidopsis). B-AZ has not previously been reported to have any biological and biochemical activities. Target identification can elucidate the mode of action of small molecules, but we were unable to make a molecular probe of B-AZ for target identification. Instead, we performed other analysis, gene expression profiling that potentially reveals mode of action of molecules. Short-term treatment of B-AZ decreased the expression of four dawn- and morning-phased clock-associated genes, CIRCADIAN CLOCK-ASSOCIATED 1 (CCA1), LATE ELONGATED HYPOCOTYL (LHY), PSEUDO-RESPONSE REGULATOR 9 (PRR9) and PRR7. Consistently, amounts of PRR5 and TIMING OF CAB EXPRESSION 1 (TOC1) proteins, transcriptional repressors of CCA1, LHY, PRR9 and PRR7 were increased upon B-AZ treatment. B-AZ inhibited Casein Kinase 1 family (CK1) that phosphorylates PRR5 and TOC1 for targeted degradation. A docking study and molecular dynamics simulation suggested that B-AZ interacts with the ATP-binding pocket of human CK1 delta, whose amino acid sequences are highly similar to those of Arabidopsis CK1. B-AZ-induced period-lengthening effect was attenuated in prr5 toc1 mutants. Collectively, this study provides a novel and simple structure CK1 inhibitor that modulates circadian clock via accumulation of PRR5 and TOC1.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Relógios Circadianos/fisiologia , Fatores de Transcrição/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Caseína Quinase I/genética , Caseína Quinase I/metabolismo , Relógios Circadianos/genética , Ritmo Circadiano/genética , Ritmo Circadiano/fisiologia , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Fatores de Transcrição/genética
15.
Plant Mol Biol ; 101(6): 521-536, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31549344

RESUMO

KEY MESSAGE: Castor patatin-like phospholipase A IIIß facilitates the exclusion of hydroxy fatty acids from phosphatidylcholine in developing transgenic Arabidopsis seeds. Hydroxy fatty acids (HFAs) are industrial useful, but their major natural source castor contains toxic components. Although expressing a castor OLEATE 12-HYDROXYLASE in Arabidopsis thaliana leads to the synthesis of HFAs in seeds, a high proportion of the HFAs are retained in phosphatidylcholine (PC). Thus, the liberation of HFA from PC seems to be critical for obtaining HFA-enriched seed oils. Plant phospholipase A (PLA) catalyzes the hydrolysis of PC to release fatty acyl chains that can be subsequently channeled into triacylglycerol (TAG) synthesis or other metabolic pathways. To further our knowledge regarding the function of PLAs from HFA-producing plant species, two class III patatin-like PLA cDNAs (pPLAIIIß or pPLAIIIδ) from castor or Physaria fendleri were overexpressed in a transgenic line of A. thaliana producing C18-HFA, respectively. Only the overexpression of RcpPLAIIIß resulted in a significant reduction in seed HFA content with concomitant changes in fatty acid composition. Reductions in HFA content occurred in both PC and TAG indicating that HFAs released from PC were not incorporated into TAG. These results suggest that RcpPLAIIIß may catalyze the removal of HFAs from PC in the developing seeds synthesizing these unusual fatty acids.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimologia , Arabidopsis/metabolismo , Ácidos Graxos/metabolismo , Fosfatidilcolinas/metabolismo , Fosfolipases/metabolismo , Plantas Geneticamente Modificadas/enzimologia , Plantas Geneticamente Modificadas/metabolismo , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Plantas Geneticamente Modificadas/genética
16.
Int J Mol Sci ; 20(18)2019 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-31505781

RESUMO

Biodiversity in plant shape is mainly attributable to the diversity of leaf shape, which is largely determined by the transient morphogenetic activity of the leaf margin that creates leaf serrations. However, the precise mechanism underlying the establishment of this morphogenetic capacity remains poorly understood. We report here that INDOLE-3-BUTYRIC ACID RESPONSE 5 (IBR5), a dual-specificity phosphatase, is a key component of leaf-serration regulatory machinery. Loss-of-function mutants of IBR5 exhibited pronounced serrations due to increased cell area. IBR5 was localized in the nucleus of leaf epidermis and petiole cells. Introducing a C129S mutation within the highly conserved VxVHCx2GxSRSx5AYLM motif of IBR5 rendered it unable to rescue the leaf-serration defects of the ibr5-3 mutant. In addition, auxin reporters revealed that the distribution of auxin maxima was expanded ectopically in ibr5-3. Furthermore, we found that the distribution of PIN1 on the plasma membrane of the epidermal and cells around the leaf vein was compromised in ibr5-3. We concluded that IBR5 is essential for the establishment of PIN-FORMED 1 (PIN1)-directed auxin maxima at the tips of leaf serration, which is vital for the elaborated regulation during its formation.


Assuntos
Proteínas de Arabidopsis/biossíntese , Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Fosfatases de Especificidade Dupla/metabolismo , Regulação da Expressão Gênica de Plantas/fisiologia , Proteínas de Membrana Transportadoras/biossíntese , Epiderme Vegetal/crescimento & desenvolvimento , Folhas de Planta/crescimento & desenvolvimento , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Fosfatases de Especificidade Dupla/genética , Proteínas de Membrana Transportadoras/genética , Mutação , Folhas de Planta/genética
17.
Int J Mol Sci ; 20(18)2019 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-31505838

RESUMO

WAX INDUCER1/SHINE1 (WIN1) belongs to the AP2/EREBP transcription factor family and plays an important role in wax and cutin accumulation in plants. Here we show that BnWIN1 from Brassica napus (Bn) has dual functions in wax accumulation and oil synthesis. Overexpression (OE) of BnWIN1 led to enhanced wax accumulation and promoted growth without adverse effects on oil synthesis under salt stress conditions. Lipid profiling revealed that BnWIN1-OE plants accumulated more waxes with elevated C29-alkanes, C31-alkanes, C28-alcohol, and C29-alcohol relative to wild type (WT) under salt stress. Moreover, overexpression of BnWIN1 also increased seed oil content under normal growth conditions. BnWIN1 directly bound to the promoter region of genes encoding biotin carboxyl carrier protein 1 (BCCP1), glycerol-3-phosphate acyltransferase 9 (GPAT9), lysophosphatidic acid acyltransferase 5 (LPAT5), and diacylglycerol acyltransferase 2 (DGAT2) involved in the lipid anabolic process. Overexpression of BnWIN1 resulted in upregulated expression of numerous genes involved in de novo fatty acid synthesis, wax accumulation, and oil production. The results suggest that BnWIN1 is a transcriptional activator to regulate the biosynthesis of both extracellular and intracellular lipids.


Assuntos
Brassica napus/metabolismo , Regulação da Expressão Gênica de Plantas/fisiologia , Lipídeos/biossíntese , Pressão Osmótica , Óleos Vegetais/metabolismo , Proteínas de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Brassica napus/genética , Lipídeos/genética , Proteínas de Plantas/genética , Regiões Promotoras Genéticas/fisiologia , Fatores de Transcrição/genética
18.
Int J Mol Sci ; 20(18)2019 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-31505875

RESUMO

he onset of leaf senescence is triggered by external cues and internal factors such as phytohormones and signaling pathways involving transcription factors (TFs). Abscisic acid (ABA) strongly induces senescence and endogenous ABA levels are finely tuned by many senescence-associated TFs. Here, we report on the regulatory function of the senescence-induced TF OsWRKY5 TF in rice (Oryza sativa). OsWRKY5 expression was rapidly upregulated in senescing leaves, especially in yellowing sectors initiated by aging or dark treatment. A T-DNA insertion activation-tagged OsWRKY5-overexpressing mutant (termed oswrky5-D) promoted leaf senescence under natural and dark-induced senescence (DIS) conditions. By contrast, a T-DNA insertion oswrky5-knockdown mutant (termed oswrky5) retained leaf greenness during DIS. Reverse-transcription quantitative PCR (RT-qPCR) showed that OsWRKY5 upregulates the expression of genes controlling chlorophyll degradation and leaf senescence. Furthermore, RT-qPCR and yeast one-hybrid analysis demonstrated that OsWRKY5 indirectly upregulates the expression of senescence-associated NAM/ATAF1/2/CUC2 (NAC) genes including OsNAP and OsNAC2. Precocious leaf yellowing in the oswrky5-D mutant might be caused by elevated endogenous ABA concentrations resulting from upregulated expression of ABA biosynthesis genes OsNCED3, OsNCED4, and OsNCED5, indicating that OsWRKY is a positive regulator of ABA biosynthesis during leaf senescence. Furthermore, OsWRKY5 expression was suppressed by ABA treatment. Taken together, OsWRKY5 is a positive regulator of leaf senescence that upregulates senescence-induced NAC, ABA biosynthesis, and chlorophyll degradation genes.


Assuntos
Ácido Abscísico/metabolismo , Regulação da Expressão Gênica de Plantas/fisiologia , Oryza/metabolismo , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Clorofila/genética , Clorofila/metabolismo , Técnicas de Silenciamento de Genes , Oryza/genética , Folhas de Planta/genética , Proteínas de Plantas/genética , Fatores de Transcrição/genética
19.
BMC Plant Biol ; 19(1): 407, 2019 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-31533618

RESUMO

BACKGROUND: NAC transcription factors contain five highly conserved subdomains which are required for protein dimerisation and DNA binding. Few residues within these subdomains have been identified as essential for protein function, and fewer still have been shown to be of biological relevance in planta. Here we use a positive regulator of senescence in wheat, NAM-A1, to test the impact of missense mutations at specific, highly conserved residues of the NAC domain on protein function. RESULTS: We identified missense mutations in five highly conserved residues of the NAC domain of NAM-A1 in a tetraploid TILLING population. TILLING lines containing these mutations, alongside synonymous and non-conserved mutation controls, were grown under glasshouse conditions and scored for senescence. Four of the five mutations showed a significant and consistent delay in peduncle senescence but had no consistent effects on flag leaf senescence. All four mutant alleles with the delayed senescence phenotype also lost the ability to interact with the homoeolog NAM-B1 in a yeast two-hybrid assay. Two of these residues were previously shown to be involved in NAC domain function in Arabidopsis, suggesting conservation of residue function between species. Three of these four alleles led to an attenuated cell death response compared to wild-type NAM-A1 when transiently over-expressed in Nicotiana benthamiana. One of these mutations was further tested under field conditions, in which there was a significant and consistent delay in both peduncle and leaf senescence. CONCLUSIONS: We combined field and glasshouse studies of a series of mutant alleles with biochemical analyses to identify four residues of the NAC domain which are required for NAM-A1 function and protein interaction. We show that mutations in these residues lead to a gradient of phenotypes, raising the possibility of developing allelic series of mutations for traits of agronomic importance. We also show that mutations in NAM-A1 more severely impact peduncle senescence, compared to the more commonly studied flag leaf senescence, highlighting this as an area deserving of further study. The results from this integrated approach provide strong evidence that conserved residues within the functional domains of NAC transcription factors have biological significance in planta.


Assuntos
Folhas de Planta/metabolismo , Folhas de Planta/fisiologia , Proteínas de Plantas/metabolismo , Triticum/metabolismo , Triticum/fisiologia , Envelhecimento , Alelos , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Proteínas de Plantas/genética , Ligação Proteica , Técnicas do Sistema de Duplo-Híbrido
20.
BMC Plant Biol ; 19(1): 397, 2019 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-31510928

RESUMO

BACKGROUND: Drought stress is a major abiotic stress that causes huge losses in agricultural production. Proso millet (Panicum miliaceum L.) can efficiently adapt to drought stress and provides important information and gene resources to improve drought tolerance. However, its complex drought-responsive mechanisms remain unclear. RESULTS: Among 37 core Chinese proso millet cultivars, Jinshu 6 (JS6) was selected as the drought-sensitive test material, whereas Neimi 5 (NM5) was selected as the drought-tolerant test material under PEG-induced water stress. After sequencing, 1695 differentially expressed genes (DEGs) were observed in JS6 and NM5 without PEG-induced water stress (JS6CK and NM5CK). A total of 833 and 2166 DEGs were found in the two cultivars under simulated drought by using 20% PEG-6000 for 6 (JS6T6 and NM5T6) and 24 h (JS6T24 and NM5T24), respectively. The DEGs in JS6T6 and JS6T24 treatments were approximately 0.298- and 0.754-fold higher than those in NM5T6 and NM5T24, respectively. Compared with the respective controls, more DEGs were found in T6 treatments than in T24 treatments. A delay in the transcriptional responses of the ROS scavenging system to simulated drought treatment and relatively easy recovery of the expression of photosynthesis-associated genes were observed in NM5. Compared with JS6, different regulation strategies were observed in the jasmonic acid (JA) signal transduction pathway of NM5. CONCLUSION: Under PEG-induced water stress, NM5 maintained highly stable gene expression levels. Compared with drought-sensitive cultivars, the different regulation strategies in the JA signal transduction pathway in drought-tolerant cultivars may be one of the driving forces underlying drought stress tolerance.


Assuntos
Secas , Regulação da Expressão Gênica de Plantas/fisiologia , Panicum/fisiologia , Transcriptoma , Panicum/genética , Folhas de Planta/genética , Folhas de Planta/fisiologia , Estresse Fisiológico
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA