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1.
J Agric Food Chem ; 67(37): 10432-10447, 2019 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-31466447

RESUMO

A composite nanogel was developed for cyanidin-3-O-glucoside (C3G) delivery by combining Maillard reaction and heat gelation. The starting materials utilized were ovalbumin, dextran, and pectin. C3G-loaded nanogel was spherical with a diameter of ∼185 nm, which was maintained over a wide range of pH and NaCl concentrations. The composite nanogel enhanced the chemical stability of C3G under accelerated degradation models and a simulated gastrointestinal tract. Clathrin-mediated, caveolae-mediated, and macropinocytosis-related endocytosis contributed to the higher cellular uptake of nano-C3G than that of free-C3G. The apparent permeability coefficients of C3G increased 2.16 times after nanoencapsulation. The transcytosis of the C3G-bearing nanogel occurred primarily through the clathrin-related pathway and macropinocytosis and followed the "common recycling endosomes-endoplasmic reticulum-Golgi complex-basolateral plasma membrane" route. Moreover, nano-C3G was more efficient in restoring the viability of cells and activities of endogenous antioxidant enzymes than free-C3G in oxidative models, which may be attributed to the former's high cellular absorption.


Assuntos
Antocianinas/química , Antocianinas/metabolismo , Antioxidantes/química , Antioxidantes/metabolismo , Portadores de Fármacos/química , Glucosídeos/química , Glucosídeos/metabolismo , Células CACO-2 , Composição de Medicamentos , Estabilidade de Medicamentos , Trato Gastrointestinal/metabolismo , Géis/química , Géis/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Absorção Intestinal , Modelos Biológicos , Nanopartículas/química , Nanopartículas/metabolismo , Tamanho da Partícula
2.
BMC Plant Biol ; 19(1): 339, 2019 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-31382883

RESUMO

BACKGROUND: Tartary buckwheat (Fagopyrum tataricum) is an edible cereal crop whose sprouts have been marketed and commercialized for their higher levels of anti-oxidants, including rutin and anthocyanin. UDP-glucose flavonoid glycosyltransferases (UFGTs) play an important role in the biosynthesis of flavonoids in plants. So far, few studies are available on UFGT genes that may play a role in tartary buckwheat flavonoids biosynthesis. Here, we report on the identification and functional characterization of seven UFGTs from tartary buckwheat that are potentially involved in flavonoid biosynthesis (and have varying effects on plant growth and development when overexpressed in Arabidopsis thaliana.) RESULTS: Phylogenetic analysis indicated that the potential function of the seven FtUFGT proteins, FtUFGT6, FtUFGT7, FtUFGT8, FtUFGT9, FtUFGT15, FtUFGT40, and FtUFGT41, could be divided into three Arabidopsis thaliana functional subgroups that are involved in flavonoid biosynthesis of and anthocyanin accumulation. A significant positive correlation between FtUFGT8 and FtUFGT15 expression and anthocyanin accumulation capacity was observed in the tartary buckwheat seedlings after cold stress. Overexpression in Arabidopsis thaliana showed that FtUFGT8, FtUFGT15, and FtUFGT41 significantly increased the anthocyanin content in transgenic plants. Unexpectedly, overexpression of FtUFGT6, while not leading to enhanced anthocyanin accumulation, significantly enhanced the growth yield of transgenic plants. When wild-type plants have only cotyledons, most of the transgenic plants of FtUFGT6 had grown true leaves. Moreover, the growth speed of the oxFtUFGT6 transgenic plant root was also significantly faster than that of the wild type. At later growth, FtUFGT6 transgenic plants showed larger leaves, earlier twitching times and more tillers than wild type, whereas FtUFGT15 showed opposite results. CONCLUSIONS: Seven FtUFGTs were isolated from tartary buckwheat. FtUFGT8, FtUFGT15, and FtUFGT41 can significantly increase the accumulation of total anthocyanins in transgenic plants. Furthermore, overexpression of FtUFGT6 increased the overall yield of Arabidopsis transgenic plants at all growth stages. However, FtUFGT15 shows the opposite trend at later growth stage and delays the growth speed of plants. These results suggested that the biological function of FtUFGT genes in tartary buckwheat is diverse.


Assuntos
Fagopyrum/genética , Genes de Plantas/genética , Glicosiltransferases/genética , Proteínas de Plantas/genética , Antocianinas/metabolismo , Arabidopsis/genética , Sequência Conservada , Fagopyrum/enzimologia , Flavonoides/metabolismo , Genes de Plantas/fisiologia , Glicosiltransferases/fisiologia , Filogenia , Proteínas de Plantas/fisiologia , Plantas Geneticamente Modificadas , Análise de Sequência de DNA
3.
Gene ; 716: 144024, 2019 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-31390541

RESUMO

The young leaves generally accumulate a certain concentration anthocyanins in the dominant species of the subtropical forest, and the changes of anthocyanin synthesis-related enzyme genes expression levels had an important effect on the study photoprotection of anthocyanins in the young leaves of subtropical forests. The determination of anthocyanin synthesis-related enzyme gene sequences and the selection of appropriate reference genes provide a basis for analyzing the functional properties of anthocyanins. In this study, four dominant subtropical forest species (i.e., Schima superba, Castanopsis fissa, Acmena acuminatissima, Cryptocarya concinna) were taken as materials. To obtain the correct nucleotide sequences of anthocyanin-related enzymes, the nucleotide sequences of CHS, DFR and ANS in each dominant species were obtained by sequencing and comparison. Then, to select the most stable reference genes for leaves at different developmental stages and different light conditions, the expression levels of six reference genes, including 18S, Actin, GAPDH, TUB, EF1 and UBQ, were studied by real-time fluorescent quantitative PCR (qRT-PCR), and reference gene stability was analyzed by GeNorm and NormFinder software. The results showed that the expression level of Actin was the most stable in S. superba, A. acuminatissima and C. concinna, and the expression level of GAPDH was the most stable in C. fissa. Finally, the expression levels of the anthocyanin synthesis genes CHS, DFR and ANS were analyzed and found to be consistent with the accumulation trend of anthocyanins in leaves. This study has important theoretical and practical significance for future research into the expression of anthocyanin synthesis-related enzyme genes in the dominant tree species in subtropical forests and reveals that anthocyanin has a photoprotective effect for young leaves in high-light environments.


Assuntos
Antocianinas/biossíntese , Árvores/genética , Aciltransferases/genética , Aciltransferases/metabolismo , Oxirredutases do Álcool/genética , Oxirredutases do Álcool/metabolismo , Antocianinas/metabolismo , Florestas , Genes de Plantas , Folhas de Planta/genética , Folhas de Planta/metabolismo , RNA de Plantas/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/normas , Padrões de Referência , Alinhamento de Sequência , Análise de Sequência , Árvores/enzimologia , Árvores/metabolismo
4.
Food Chem ; 300: 125169, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31336273

RESUMO

Red drupelet is a postharvest disorder of blackberries with several drupelets turning back to red. This affects visual quality and thus marketability and consumers' acceptance. However, the cause of this disorder as well as metabolite changes during color reversion have not been fully understood. Anthocyanins, cyanidin 3-glucoside, cyanidin 3-malonylglucoside, cyanidin 3-dioxalylglucoside, and total anthocyanin, were significantly lower in red drupelets than in black drupelets after 7 days of storage. Sugars and organic acids, lipids, and free amino acids also changed with storage and by color reversion. The untargeted metabolomics analyses indicated that red drupelets were generally differentiated from berries at harvest or black drupelets at metabolite level. The results of this study help better understand the red drupelet disorder. To our knowledge, this is the first study investigating red drupelet disorder by comparing black and red drupelets at metabolite level.


Assuntos
Metabolômica/métodos , Rubus/metabolismo , Aminoácidos/análise , Aminoácidos/metabolismo , Antocianinas/análise , Antocianinas/metabolismo , Cor , Qualidade dos Alimentos , Armazenamento de Alimentos , Frutas/química , Glucosídeos/análise , Glucosídeos/metabolismo , Lipídeos/análise , Rubus/química
5.
Food Chem ; 298: 124973, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31261005

RESUMO

The aim of the present study was to explore the underlying mechanisms involved in anthocyanin biosynthesis in purple, blue, and white barley using quantitative proteomics analysis. We identified the differences in protein expression and related functions involved in anthocyanin biosynthesis in purple, blue, and white barley (named H, M, and L groups, respectively, based on their anthocyanin content) using TMT-liquid chromatography/mass spectroscopy-based proteomic methods. Totally, 297, 300, 254, and 1421 differentially expressed proteins (DEPs) were found in H vs. L, H vs. M, L vs. M, and H vs. L vs. M groups, respectively. Six clusters of proteins from the 1421 DEPs were mainly involved in carbon metabolism, amino acid and secondary metabolite biosynthesis, and metabolic pathways. Several proteins were validated using parallel reaction monitoring. The proteins involved in amino acid biosynthesis, carbon metabolism, metabolic pathways, and phenylpropanoid biosynthesis were responsible for the color differences in the three barley varieties.


Assuntos
Antocianinas/metabolismo , Hordeum/metabolismo , Proteínas de Plantas/análise , Proteínas de Plantas/metabolismo , Cromatografia Líquida , Pigmentação , Proteômica/métodos , Espectrometria de Massas em Tandem
6.
Sci Total Environ ; 671: 466-473, 2019 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-31331442

RESUMO

Removal of bio-accumulated pesticides in edible fish is a global problem. In this study, we tested protective capability of a phytochemical pelargonidin-loaded non-toxic, biodegradable poly-lactide-co-glycolide nano-particles (NPG) against toxicity induced by a pesticide cypermethrin (CM) in a fish model (Oreochromis mossambica) in vivo and also in L6 muscle cell line, in vitro. First we assessed potential sustainable release of nanoparticles following oral administration of NPG to fish, their ability to cross sub-cellular membranes in several tissues and efficacy to cross blood-brain-barrier. Next, protective ability of NPG, if any, against CM in fish was evaluated deploying parameters like % cell viability, DNA damage in muscle cells and modulation of anti-oxidative-enzymes like superoxide dismutase, catalase and lipid peroxidase. Modulation of reactive oxygen species generation, nuclear condensation and alteration in stress related protein signalling cascade were assessed in L6 cells. Results revealed that NPG had nano-size range (~10-12 nm) and negative zeta potential (-17 mV). Bioavailability and distribution of NPG could be followed by spectrophotometric absorbance of pelargonidin at 293 nm from 6 h onward till 24 h in all important tissues including the brain. Thus, 0.5 mg/g b.w. NPG could demonstrate protective ability in CM-intoxicated fish muscle cells in respect of % cell viability, DNA damage and stress related enzymes. Similar alterations could also be found in signalling protein cascade in L6 cells in response to treatment of 5 µg/ml NPG against CM-induced toxicity and depletion of overall ROS generation and nuclear condensation. Therefore, NPG could be used as a potential drug in management of pesticide toxicity in cultured edible fish.


Assuntos
Antocianinas/metabolismo , Antioxidantes/metabolismo , Ciclídeos/fisiologia , Praguicidas/toxicidade , Poluentes Químicos da Água/toxicidade , Animais , Catalase/metabolismo , Dano ao DNA , Estresse Oxidativo , Transdução de Sinais , Superóxido Dismutase/metabolismo
7.
BMC Plant Biol ; 19(1): 315, 2019 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-31307378

RESUMO

BACKGROUND: Abaxially anthocyanic leaves of deeply-shaded understorey plants play important ecological significance for the environmental adaption. In contrast to the transient pigmentation in other plants, anthocyanins are permanently presented in these abaxially red leaves, however, the mechanism for the pigment maintenance remains unclear. In the present study, we investigated phenolic metabolites that may affect pigment stability and degradation in Excoecaria cochinchinensis (a bush of permanently abaxial-red leaves), via a comparison with Osmanthus fragrans (a bush of transiently red leaves). RESULTS: High levels of galloylated anthocyanins were identified in the Excoecaria but not in the Osmanthus plants. The galloylated anthocyanin showed slightly higher stability than two non-galloylated anthocyanins, while all the 3 pigments were rapidly degraded by peroxidase (POD) in vitro. High levels of hydrolysable tannins [mainly galloylglucoses/ellagitannins (GGs/ETs)] were identified in Excoecaria but none in Osmanthus. GGs/ETs showed inhibition effect on POD, with IC50 ranged from 35.55 to 83.27 µM, correlated to the markedly lower POD activities detected in Excoecaria than in Osmanthus. Strong copigmentation was observed for GGs/ETs and anthocyanins, with more than 30% increase in the red intensity of non-galloylated anthocyanin solutions. In the leaf tissue, the hydrolysable tannins were observed to be co-localized with anthocyanins at the abaxial layer of the Excoecaria leaves, correlated to the low POD activity, more acidity and increased red intensity of the tissue. CONCLUSION: The results suggest that the Excoecaria leaves accumulate a distinct group of phenolic metabolites, mainly GGs/ETs, at the abaxial layer, which prevent anthocyanin degradation and increase the pigment stability, and consequently lead to the permanent maintenance of the red leaves.


Assuntos
Antocianinas/metabolismo , Euphorbiaceae/metabolismo , Taninos Hidrolisáveis/metabolismo , Peroxidase/antagonistas & inibidores , Pigmentação , Folhas de Planta/metabolismo , Euphorbiaceae/enzimologia , Oleaceae/metabolismo , Peroxidase/metabolismo , Folhas de Planta/crescimento & desenvolvimento
8.
Plant Cell Rep ; 38(8): 927-936, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31147728

RESUMO

KEY MESSAGE: A new anthocyanin biosynthesis transcription factor PdMYB118, which could be used for the genetic engineering of colorful tree species, was indentified from a red leaf mutant of Populus deltoids. In higher plants, the biosynthesis of anthocyanins is regulated by several classes of transcription factors (TFs), including R2R3-MYB, bHLH and WD-repeat proteins. In this work, we isolated an MYB gene regulating anthocyanin biosynthesis from a red leaf mutant of Populus deltoids, which accumulated more anthocyanins in the leaves and showed higher expression levels of anthocyanin biosynthesis genes than did the wild type. Gene expression analyses of all TFs regulating anthocyanin biosynthesis demonstrated that only a MYB118 homologous gene, PdMYB118, was up-regulated in the mutant compared with the wide type. Subcellular localization analyses in poplar leaf mesophyll protoplasts showed that PdMYB118-YFP fusion protein was specifically located in nucleus. When transiently expressed in poplar leaf protoplasts, PdMYB118 specifically promoted the expression of anthocyanidin biosynthesis genes. Dual-luciferase assays revealed that PdMYB118 can directly activate the promoters of these genes. When overexpressed in Shanxin Yang (P. davidiana × P. bolleana), a hybrid clone commercially grown for landscaping in the northern part of China, transgenic plants overexpressing PdMYB118 produced more anthocyanins in the leaves and turned their color into redness when grown in both greenhouse and field. Consistently, transcripts of some important anthocyanidin biosynthesis genes were significantly increased in the leaves of transgenic plants. All these results indicate that PdMYB118 functions as an essential transcription factor regulating anthocyanin biosynthesis in poplar and could be used for the genetic engineering of colorful tree species.


Assuntos
Antocianinas/metabolismo , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Populus/metabolismo , Fatores de Transcrição/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Mutação/genética , Folhas de Planta/genética , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética , Populus/genética , Fatores de Transcrição/genética
9.
Food Chem ; 293: 57-65, 2019 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-31151649

RESUMO

The bioavailability of drugs can be improved by regulating the structural properties, particularly lipoid systems, such as niosomes, can increase cellular uptake. Herein, we optimized double emulsion and niosomal formulations for encapsulating anthocyanin-rich black carrot extract. Nanoparticles obtained by selected formulation were characterized in terms of morphology, particle size, drug encapsulation efficiency, in vitro release and cytotoxicity. The optimum conditions for niosomal formulation were elicited as 30 mg of cholesterol, 150 mg of Tween 20 and feeding time of 1 min at a stirring rate of 900 rpm yielding the lowest average particle size of 130 nm. In vitro release data showed the majority of the encapsulated anthocyanins were released at the end of 10 h. A mathematical model was developed to estimate the absorption of anthocyanins released from niosomes and cytotoxicity was assessed against neuroblastoma. Overall, these findings suggest that niosomal vesicles might be suitable delivery systems for anthocyanins.


Assuntos
Antocianinas/química , Lipossomos/química , Antocianinas/metabolismo , Varredura Diferencial de Calorimetria , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Colesterol/química , Daucus carota/química , Daucus carota/metabolismo , Humanos , Lipossomos/toxicidade , Modelos Teóricos , Nanopartículas/química , Tamanho da Partícula , Polissorbatos/química
10.
Food Chem ; 295: 607-612, 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31174802

RESUMO

The purpose of this study is to investigate the effects of sodium nitroprusside (SNP) treatment after harvest on the activity of antioxidative enzymes and the phenylpropanoid pathway of blueberries. Blueberry fruits were dipped in 1.0 mmol/L SNP solution for 10 min and stored at 4 °C. Fruits treated with distilled water were used as the control. The results indicated that SNP significantly inhibited the increase of weight loss and enhanced the ascorbic acid content of blueberry fruit. Moreover, SNP increased the activity of phenylalanine ammonia-lyase, 4-coumarate-CoA ligase, polyphenol oxidase, superoxide dismutase, glutathione reductase, ascorbate peroxidase, peroxidase, and hydrogen peroxide in blueberry fruit. The accumulation of lignin and anthocyanin in the fruit was also stimulated by the SNP treatment. These results demonstrate that SNP treatment could maintain the antioxidant ability of blueberries by regulating the phenylpropanoid pathway and antioxidant enzymes.


Assuntos
Antioxidantes/metabolismo , Mirtilos Azuis (Planta)/efeitos dos fármacos , Nitroprussiato/farmacologia , Antocianinas/metabolismo , Antioxidantes/química , Ascorbato Peroxidases/metabolismo , Ácido Ascórbico/metabolismo , Mirtilos Azuis (Planta)/química , Mirtilos Azuis (Planta)/metabolismo , Frutas/química , Frutas/efeitos dos fármacos , Frutas/metabolismo , Glutationa Redutase/metabolismo , Peróxido de Hidrogênio/metabolismo , Lignina/metabolismo , Peroxidase/metabolismo , Superóxido Dismutase/metabolismo
11.
BMC Plant Biol ; 19(1): 263, 2019 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-31215400

RESUMO

BACKGROUND: Because flavonoids and trichomes play crucial roles in plant defence, their formation requires fine transcriptional control by multiple transcription factor families. However, little is known regarding the mechanism of the R2R3-MYB transcription factors that regulate both flavonoid metabolism and trichome development. RESULTS: Here, we identified a unique SG4-like-MYB TF from Tartary buckwheat, FtMYB8, which harbours the C2 repression motif and an additional TLLLFR repression motif. The expression profiles of FtMYB8 combined with the transcriptional activity of PFtMYB8 promoter showed that FtMYB8 mRNA mainly accumulated in roots during the true leaf stage and flowering stage and in bud trichomes and flowers, and the expression of this gene was markedly induced by MeJA, ABA and UV-B treatments but repressed by dark treatment. Overexpression of FtMYB8 in Arabidopsis reduces the accumulation of anthocyanin/proanthocyanidin by specifically inhibiting TT12 expression, which may depend on the interaction between FtMYB8 and TT8. Interestingly, this interaction may also negatively regulate the marginal trichome initiation in Arabidopsis leaves. CONCLUSIONS: Taken together, our results suggest that FtMYB8 may fine-tune the accumulation of anthocyanin/proanthocyanidin in the roots and flowers of Tartary buckwheat by balancing the inductive effects of transcriptional activators, and probably regulate trichome distribution in the buds of Tartary buckwheat.


Assuntos
Antocianinas/metabolismo , Fagopyrum/metabolismo , Proteínas de Plantas/metabolismo , Proantocianidinas/metabolismo , Fatores de Transcrição/metabolismo , Tricomas/crescimento & desenvolvimento , Arabidopsis , Fagopyrum/genética , Fagopyrum/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/fisiologia , Plantas Geneticamente Modificadas , Alinhamento de Sequência , Fatores de Transcrição/genética , Fatores de Transcrição/fisiologia , Transcriptoma , Tricomas/metabolismo
12.
Plant Mol Biol ; 100(4-5): 451-465, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31079310

RESUMO

KEY MESSAGE: AcGST1, an anthocyanin-related GST, may functions as a carrier to transport anthocyanins from ER to tonoplast in kiwifruit. It was positively regulated by AcMYBF110 through directly binding to its promoter. Anthocyanins are synthesized in the cytoplasmic surface of the endoplasmic reticulum but accumulate predominantly in the vacuole. Previous studies in model and ornamental plants have suggested that a member of the glutathione S-transferase (GST) gene family is involved in sequestration of anthocyanins into the vacuole. However, little is known about anthocyanin-related GST protein in kiwifruit. Here, four putative AcGSTs were identified from the genome of the red-fleshed Actinidia chinensis cv 'Hongyang'. Expression analyses reveal only the expression of AcGST1 was highly consistent with anthocyanin accumulation. Molecular complementation of Arabidopsis tt19 demonstrates AcGST1 can complement the anthocyanin-less phenotype of tt19. Transient expression in Actinidia arguta fruits further confirms that AcGST1 is functional in anthocyanin accumulation in kiwifruit. In vitro assays show the recombinant AcGST1 increases the water solubility of cyanidin-3-O-galactoside (C3Gal) and cyanidin-3-O-xylo-galactoside (C3XG). We further show that AcGST1 protein is localized not only in the ER but also on the tonoplast, indicating AcGST1 (like AtTT19) may functions as a carrier protein to transport anthocyanins to the tonoplast in kiwifruit. Moreover, the promoter of AcGST1 can be activated by AcMYBF110, based on results from transient dual-luciferase assays and yeast one-hybrid assays. EMSAs show that AcMYBF110 binds directly to CAGTTG and CCGTTG motifs in the AcGST1 promoter. These results indicate that AcMYBF110 plays an important role in transcriptional regulation of AcGST1 and, therefore, in controlling accumulation of anthocyanins in kiwifruit.


Assuntos
Actinidia/genética , Antocianinas/metabolismo , Glutationa Transferase/genética , Proteínas de Plantas/genética , Actinidia/enzimologia , Actinidia/metabolismo , Transporte Biológico , Clonagem Molecular , Retículo Endoplasmático/metabolismo , Frutas/enzimologia , Frutas/genética , Frutas/metabolismo , Regulação da Expressão Gênica de Plantas , Genoma de Planta , Glutationa Transferase/química , Glutationa Transferase/fisiologia , Proteínas de Plantas/química , Proteínas de Plantas/fisiologia , Regiões Promotoras Genéticas
13.
Plant Physiol Biochem ; 141: 60-72, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31128564

RESUMO

The MBW complex, consisting of MYB, basic helix-loop-helix (bHLH) and WD40 proteins, regulates multiple traits in plants, such as anthocyanin and proanthocyanidin biosynthesis and cell fate determination. The complex has been widely identified in dicot plants, whereas few studies are concentrated on monocot plants which are of crucial importance to decipher its functional diversities among angiosperms during evolution. In present study, a WD40 gene from Freesia hybrida, designated as FhTTG1, was cloned and functionally characterized. Real-time PCR analysis indicated that it was expressed synchronously with the accumulation of both proanthocyanidins and anthocyanins in Freesia flowers. Transient protoplast transfection and biomolecular fluorescence complementation (BiFC) assays demonstrated that FhTTG1 could interact with FhbHLH proteins (FhTT8L and FhGL3L) to constitute the MBW complex. Moreover, the transportation of FhTTG1 to nucleus was found to rely on FhbHLH factors. Outstandingly, FhTTG1 could highly activate the anthocyanin or proanthocyanidin biosynthesis related gene promoters when co-transfected with MYB and bHLH partners, implying that FhTTG1 functioned as a member of MBW complex to control the anthocyanin or proanthocyanidin biosynthesis in Freesia hybrida. Further ectopic expression assays in Arabidopsis ttg1-1 showed the defective phenotypes of ttg1-1 were partially restored. Molecular biological assays validated FhTTG1 might interact with the endogenous bHLH factors to up-regulate genes responsible for anthocyanin and proanthocyanidin biosynthesis and trichome formation, indicating that FhTTG1 might perform exchangeable roles with AtTTG1. These results will not only contribute to the characterization of FhTTG1 in Freesia but also shed light on the establishment of flavonoid regulatory system in monocot plants, especially in Freesia hybrida.


Assuntos
Antocianinas/metabolismo , Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Iridaceae/metabolismo , Proantocianidinas/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Flores/genética , Flores/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Iridaceae/genética , Mutação , Filogenia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Protoplastos/metabolismo , Tricomas/metabolismo , Repetições WD40
14.
BMC Genomics ; 20(1): 343, 2019 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-31064320

RESUMO

BACKGROUND: Caixin and Zicaitai (Brassica rapa) belong to Southern and Central China respectively. Zicaitai contains high amount of anthocyanin in leaf and stalk resulting to the purple color. Stalk is the major edible part and stalk color is an economically important trait for the two vegetables. The aim of this study is to construct a high density genetic map using the specific length amplified fragment sequencing (SLAF-seq) technique to explore genetic basis for anthocyanin pigmentation traits via quantitative trait loci (QTL) mapping. RESULTS: We constructed a high generation linkage map with a mapping panel of F2 populations derived from 150 individuals of parental lines "Xianghongtai 01" and "Yinong 50D" with purple and green stalk respectively. The map was constructed containing 4253 loci, representing 10,940 single nucleotide polymorphism (SNP) markers spanning 1030.04 centiMorgans (cM) over 10 linkage groups (LGs), with an average distance between markers of 0.27 cM. Quantitative trait loci (QTL) analysis revealed that a major locus on chromosome 7 and 4 minor QTLs explaining 2.69-61.21% of phenotypic variation (PVE) were strongly responsible for variation in stalk color trait. Bioinformatics analysis of the major locus identified 62 protein-coding genes. Among the major locus, there were no biosynthetic genes related to anthocyanin. However, there were several transcription factors like helix-loop-helix (bHLH) bHLH, MYB in the locus. Seven predicted candidate genes were selected for the transcription level analysis. Only bHLH49 transcription factor, was significantly higher expressed in both stalks and young leaves of Xianghongtai01 than Yinong50D. An insertion and deletion (InDel) marker developed from deletion/insertion in the promoter region of bHLH49 showed significant correlation with the stalk color trait in the F2 population. CONCLUSION: Using the constructed high-qualified linkage map, this study successfully identified QTLs for stalk color trait. The identified valuable markers and candidate genes for anthocyanin accumulation in stalk will provide useful information for molecular regulation of anthocyanin biosynthesis. Overall our findings will lay a foundation for functional gene cloning, marker-assisted selection (MAS) and molecular breeding of important economic traits in B. rapa.


Assuntos
Antocianinas/metabolismo , Brassica rapa/anatomia & histologia , Brassica rapa/genética , Cromossomos de Plantas , Locos de Características Quantitativas , Brassica rapa/crescimento & desenvolvimento , Mapeamento Cromossômico , Ligação Genética , Marcadores Genéticos , Técnicas de Genotipagem , Fenótipo , Pigmentação , Análise de Sequência de DNA
15.
J Agric Food Chem ; 67(22): 6241-6247, 2019 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-31117508

RESUMO

A question in cell culture and dietary studies on protection by flavonoids against conditions such as hepatocyte steatosis is whether effects might be due to phenolic breakdown/digestion products. In HepG2 hepatocytes, treatment with quercetin, cyanidin, or their phenolic breakdown/digestion products (protocatechuic acid, 2,4,6-trihydroxybenzaldehyde, and caffeic acid), starting 2 h prior to oleic acid for 24 h, protected similarly against increases in intracellular lipid and reactive oxygen species and decreased mitochondrial membrane potential. Cyanidin or the phenolic products also protected against decreased mitochondrial content. After preincubation for only 1 h (to limit spontaneous degradation) and removal prior to oleic acid, only the phenolic products protected against decreased mitochondrial content, and without adding oleic acid, only protocatechuic acid and caffeic acid, and less so cyanidin, induced mitochondrial content. The results suggest that phenolic breakdown/digestion products of cyanidin and quercetin contribute to the protective effects in vitro, and perhaps in vivo.


Assuntos
Antocianinas/farmacologia , Fígado Gorduroso/tratamento farmacológico , Mitocôndrias/efeitos dos fármacos , Fenóis/metabolismo , Quercetina/farmacologia , Antocianinas/química , Antocianinas/metabolismo , Fígado Gorduroso/metabolismo , Flavonoides/química , Flavonoides/metabolismo , Flavonoides/farmacologia , Células Hep G2 , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Humanos , Mitocôndrias/metabolismo , Modelos Biológicos , Estresse Oxidativo/efeitos dos fármacos , Fenóis/química , Fenóis/farmacologia , Quercetina/química , Quercetina/metabolismo , Espécies Reativas de Oxigênio/metabolismo
16.
Plant Physiol Biochem ; 140: 88-95, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31085450

RESUMO

Although melatonin effects on postharvest fruit ripening have been studied in some detail, information is still scarce during pre-harvest. Here, we examined whether or not melatonin may exert a regulatory role during sweet cherries ripening in orchard trees. We evaluated (i) the endogenous variations in melatonin contents, in comparison to those of well-known phytohormones such as ABA, salicylic acid and jsamonic acid, by ultrahigh performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) during fruit ripening over two consecutive years, and (ii) to what extent melatonin treatments at low and high concentrations (at 10-4 M and 10-5 M, respectively) influence fruit ripening on the tree. Endogenous melatonin contents decreased in parallel to those of salicylic acid and jasmonic acid, while ABA contents increased as fruit ripening progressed, thus suggesting an inhibitory role for melatonin in fruit ripening. Furthermore, melatonin treatment at 10-5 M, which transiently increased endogenous melatonin contents at physiological concentrations, delayed anthocyanin accumulation, thus confirming an inhibitory regulatory role for melatonin in fruit ripening. We also found that the endogenous contents of cytokinins, but not those of ABA were transiently affected by melatonin treatment at 10-5 M. It is concluded that melatonin may delay sweet cherries ripening in orchard trees, probably exerting a modulatory role through a hormonal cross-talk. These results have important implications for the use of melatonin in the control of the timing of sweet cherries ripening in orchard trees.


Assuntos
Melatonina/farmacologia , Prunus avium/efeitos dos fármacos , Prunus avium/metabolismo , Antocianinas/metabolismo , Frutas/efeitos dos fármacos , Frutas/metabolismo , Espectrometria de Massas em Tandem
17.
Plant Sci ; 283: 202-210, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31128690

RESUMO

Salinity is one of the most frequent limiting conditions in pasture production for grazing livestock. Legumes, such as Lotus spp. with high forage quality and capable of adapting to different environments, improves pasture performance in restrictive areas. In order to determine potential cultivars with better forage traits, the current study assess the response to salt stress of L. tenuis, L. corniculatus and a novel L. tenuis x L. corniculatus accession. For this purpose, chlorophyll fluorescence, biomass production, ion accumulation and anthocyanins and proanthocyanidins levels have been evaluated in control and salt-treated plants PSII activity was affected by salt in L. tenuis, but not in L. corniculatus or hybrid plants. Analyzed accessions showed similar values of biomass, Na+ and K+ levels after salt treatment. Increasing Cl- concentrations were observed in all accessions. However, hybrid plants accumulate Cl- in stems at higher levels than their parental. At the same time, the levels of anthocyanins considerably increased in L. tenuis x L. corniculatus stems. Chloride and anthocyanin accumulation in stems could explain the best performance of hybrid plants after a long saline treatment. Finally, as proanthocyanidins levels were no affected by salt, L. tenuis x L. corniculatus plants maintained adequate levels to be used as ruminant feed. In conclusion, these results suggest that hybrid plants have a high potential to be used as forage on salt-affected lands. High Cl- and anthocyanins accumulation in Lotus spp. stems seems to be a trait associated to salinity tolerance, with the possibility of being used in legume breeding programs.


Assuntos
Lotus/metabolismo , Antocianinas/metabolismo , Clorofila/metabolismo , Hibridização Genética , Lotus/crescimento & desenvolvimento , Lotus/fisiologia , Melhoramento Vegetal/métodos , Potássio/metabolismo , Estresse Salino , Plantas Tolerantes a Sal , Sódio/metabolismo
18.
Plant Sci ; 283: 266-277, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31128697

RESUMO

Hormones play an important role in fruit ripening and in response to biotic stress. Nevertheless, analyses of hormonal profiling during plant development and defense are scarce. In this work, changes in hormonal metabolism in grapevine (Vitis vinifera) were compared between a susceptible (Trincadeira) and a tolerant (Syrah) variety during grape ripening and upon infection with Botrytis cinerea. Infection of grapes with the necrotrophic pathogen Botrytis cinerea leads to significant economic losses worldwide. Peppercorn-sized fruits were infected in the field and mock-treated and infected berries were collected at green, veraison and harvest stages for hormone analysis and targeted qPCR analysis of genes involved in hormonal metabolism and signaling. Results indicate a substantial reprogramming of hormonal metabolism during grape ripening and in response to fungal attack. Syrah and Trincadeira presented differences in the metabolism of abscisic acid (ABA), indole-3-acetic acid (IAA) and jasmonates during grape ripening that may be connected to fruit quality. On the other hand, high basal levels of salicylic acid (SA), jasmonates and IAA at an early stage of ripening, together with activated SA, jasmonates and IAA signaling, likely enable a fast defense response leading to grape resistance/ tolerance towards B. cinerea. The balance among the different phytohormones seems to depend on the ripening stage and on the intra-specific genetic background and may be fundamental in providing resistance or susceptibility. In addition, this study indicated the involvement of SA and IAA in defense against necrotrophic pathogens and gains insights into possible strategies for conventional breeding and/or gene editing aiming at improving grape quality and grape resistance against Botrytis cinerea.


Assuntos
Ácido Abscísico/metabolismo , Botrytis/metabolismo , Ciclopentanos/metabolismo , Ácidos Indolacéticos/metabolismo , Oxilipinas/metabolismo , Doenças das Plantas/microbiologia , Reguladores de Crescimento de Planta/metabolismo , Ácido Salicílico/metabolismo , Vitis/metabolismo , Ácido Abscísico/fisiologia , Antocianinas/metabolismo , Reguladores de Crescimento de Planta/fisiologia , Reação em Cadeia da Polimerase em Tempo Real , Especificidade da Espécie , Vitis/crescimento & desenvolvimento , Vitis/microbiologia
19.
Plant Sci ; 283: 32-40, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31128702

RESUMO

Light is an important environmental factor affecting plant growth and development. Additionally, HY5 is a central factor that coordinates light signal transduction and regulates the expression of flower color-related genes. However, there are few reports describing the co-regulation of apple fruit coloration by MdHY5 and MYB transcription factors. In this study, we detected a light-inducible gene, MdMYBDL1, which encodes a MYB-like domain and is homologous to AtMYBD in Arabidopsis thaliana. Moreover, we observed that MdHY5 binds to the G-box element of the MdMYBDL1 promoter to upregulate expression. The overexpression of MdMYBDL1 enhanced anthocyanin accumulation in apple calli and inhibited the expression of MdMYB16 and its homolog, MdMYB308. Furthermore, MdMYB16 can form a dimer with MdMYB308 and functions as a negative regulator of anthocyanin biosynthesis. Interestingly, MdMYB16 and MdMYB308 promoter activities were inhibited by MdMYBDL1 and MdHY5. These findings imply that MdHY5 responds to light signals and functions upstream of different types of MYB transcription factors, ultimately regulating anthocyanin accumulation in apples.


Assuntos
Antocianinas/metabolismo , Malus/metabolismo , Proteínas de Plantas/fisiologia , Fatores de Transcrição/fisiologia , Imunoprecipitação da Cromatina , Ensaio de Desvio de Mobilidade Eletroforética , Glucuronidase/metabolismo , Proteínas de Plantas/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Transcrição/metabolismo , Técnicas do Sistema de Duplo-Híbrido
20.
Plant Sci ; 283: 41-50, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31128712

RESUMO

Litchi (Litchi chinensis Sonn.) is a subtropical fruit known for its attractive red pericarp color, semi-translucent white aril and unique flavor and aroma. Rapid post-harvest pericarp browning strictly limits litchi fruit marketing. In the current research, we hypothesized that modification of litchi fruit pericarp anatomy by hormone application may reduce fruit susceptibility to post-harvest pericarp browning. In this context, we hypothesized that cytokinin treatment, known to induce cell division, may yield fruit with thicker pericarp and reduced susceptibility for fruit surface micro-crack formation, water loss and post-harvest pericarp browning. Exogenous cytokinin treatment was applied at different stages along the course of litchi fruit development and the effect on fruit pericarp anatomy, fruit maturation and postharvest pericarp browning was investigated. Interestingly, cytokinin treatment, applied 4 weeks after full female bloom (WFB), during the phase of pericarp cell division, led to mature fruit with thicker pericarp, reduced rate of post-harvest water loss and reduced susceptibility to post-harvest pericarp browning, as compared to non-treated control fruit. Histological sections ascribe the difference in pericarp anatomy to increased cell proliferation in the parenchymatic tissue and the highly-lignified brachysclereid cell layer. In contrast, exogenous cytokinin treatment applied 7 WFB, following the phase of pericarp cell division, significantly increased epidermal-cell proliferation but had no significant effect on overall fruit pericarp thickness and only minor affect on post-harvest water loss or pericarp browning. Interestingly, the late cytokinin treatment also significantly postponed fruit maturation-associated anthocyanin accumulation and chlorophyll degradation, as previously reported, but had no effect on other parameters of fruit maturation, like total soluble sugars and total titratable acids typically modified during aril maturation. In conclusion, exogenous cytokinin treatment at different stages in fruit development differentially modifies litchi fruit pericarp anatomy by induction of cell-type specific cell proliferation. Early cytokinin treatment during the phase of pericarp cell division may prolong litchi fruit storage by reducing fruit susceptibility to post-harvest water loss and pericarp browning.


Assuntos
Citocininas/farmacologia , Resistência à Doença/efeitos dos fármacos , Frutas/efeitos dos fármacos , Litchi/efeitos dos fármacos , Antocianinas/metabolismo , Clorofila/metabolismo , Produção Agrícola/métodos , Frutas/anatomia & histologia , Frutas/crescimento & desenvolvimento , Litchi/anatomia & histologia , Litchi/crescimento & desenvolvimento
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