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1.
Plant Mol Biol ; 104(3): 309-325, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32833148

RESUMO

KEY MESSAGE: FtMYB18 plays a role in the repression of anthocyanins and proanthocyanidins accumulation by strongly down-regulating the CHS and DFR genes in Tartary buckwheat, and the C5 motif plays an important role in this process. Anthocyanins and proanthocyanidins (PAs) are important flavonoids in Tartary buckwheat (Fagopyrum tataricum Gaertn.), which provides various vibrant color and stronge abiotic stress resistance. Their synthesis is generally regulated by MYB transcription factors at transcription level. However, the negative regulations of MYB and their effects on flavonol metabolism are poorly understood. A SG4-like MYB subfamily TF, FtMYB18, containing C5 motif was identified from Tartary buckwheat. The expression of FtMYB18 was not only showed a negative correlation with anthocyanins and PAs content but also strongly respond to MeJA and ABA. As far as the transgenic lines with FtMYB18 overexpression, anthocyanins and PAs accumulations were decreased through down-regulating expression levels of NtCHS and NtDFR in tobacco, AtDFR and AtTT12 in Arabidopsis, FtCHS, FtDFR and FtANS in Tartary buckwheat hairy roots, respectively. However, FtMYB18 showed no effect on the FLS gene expression and the metabolites content in flavonol synthesis branch. The further molecular interaction analysis indicated FtMYB18 could mediate the inhibition of anthocyanins and PAs synthesis by forming MBW transcriptional complex with FtTT8 and FtTTG1, or MYB-JAZ complex with FtJAZ1/-3/-4/-7. Importantly, in FtMYB18 mutant lines with C5 motif deletion (FtMYB18-C), both of anthocyanins and PAs accumulations had recovered to the similar level as that in wild type, which was attributed to the weakened MBW complex activity or the deficient molecular interaction between FtMYB18ΔC5 with FtJAZ3/-4. The results showed that FtMYB18 could suppress anthocyanins and PAs synthesis at transcription level through the specific interaction of C5 motif with other proteins in Tartary buckwheat.


Assuntos
Antocianinas/biossíntese , Fagopyrum/metabolismo , Proteínas de Plantas/metabolismo , Proantocianidinas/biossíntese , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Arabidopsis , Fagopyrum/genética , Flavonoides/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Estresse Fisiológico , Tabaco/genética , Fatores de Transcrição/química
2.
Gene ; 760: 144990, 2020 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-32721476

RESUMO

The MYB transcription factors are involved in the regulation of plant secondary metabolism, cell development and morphogenesis, and stress response. Here, a full-length, 816-bp NtMYB4a cDNA, which encodes a protein comprising 271 amino acids, was isolated from tobacco leaves. Phylogenetic analysis revealed that NtMYB4a is most similar to Nicotiana. attenuata MYB4, followed by Eriobotrya japonica MYB4, and NtMYB4a clustered with transcriptional activators rather than repressors. Subcellular localization assays showed that NtMYB4 localized in the nucleus, membrane, and cytoplasm. Expression analyses revealed differential expression of NtMYB4a among different tissues and organs and between different developmental stages, with most expression occurring in the stems and leaves during the full-bloom stage. Moreover, NtMYB4a expression was induced by cold, NaCl, PEG, abscisic acid, methyl jasmonate, and dark stressors, and the expression patterns and maximum expression levels varied with the type of stress. Overexpression of NtMYB4a upregulated NtPAL, Nt4CL, NtCHS, NtCHI, NtF3H, NtDFR, NtANS, and NtUFGT, which resulted in increased anthocyanin content in the tobacco corolla and darker colors. However, CRISPR/Cas9-mediated knockout of NtMYB4a downregulated NtPAL, NtC4H, Nt4CL, NtCHS, NtCHI, NtF3H, NtANS, and NtUFGT, which resulted in reduced anthocyanin content, and lighter corolla colors. These results indicated that NtMYB4a positively regulates anthocyanin biosynthesis and is involved in abiotic stress responses in tobacco plants.


Assuntos
Tabaco/metabolismo , Fatores de Transcrição/isolamento & purificação , Fatores de Transcrição/metabolismo , Ácido Abscísico/metabolismo , Sequência de Aminoácidos , Antocianinas/biossíntese , Regulação da Expressão Gênica de Plantas/genética , Filogenia , Folhas de Planta/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Estresse Fisiológico/genética , Tabaco/genética , Fatores de Transcrição/genética , Ativação Transcricional/genética
3.
Nature ; 583(7815): 277-281, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32528176

RESUMO

Plant hormones known as strigolactones control plant development and interactions between host plants and symbiotic fungi or parasitic weeds1-4. In Arabidopsis thaliana and rice, the proteins DWARF14 (D14), MORE AXILLARY GROWTH 2 (MAX2), SUPPRESSOR OF MAX2-LIKE 6, 7 and 8 (SMXL6, SMXL7 and SMXL8) and their orthologues form a complex upon strigolactone perception and play a central part in strigolactone signalling5-10. However, whether and how strigolactones activate downstream transcription remains largely unknown. Here we use a synthetic strigolactone to identify 401 strigolactone-responsive genes in Arabidopsis, and show that these plant hormones regulate shoot branching, leaf shape and anthocyanin accumulation mainly through transcriptional activation of the BRANCHED 1, TCP DOMAIN PROTEIN 1 and PRODUCTION OF ANTHOCYANIN PIGMENT 1 genes. We find that SMXL6 targets 729 genes in the Arabidopsis genome and represses the transcription of SMXL6, SMXL7 and SMXL8 by binding directly to their promoters, showing that SMXL6 serves as an autoregulated transcription factor to maintain the homeostasis of strigolactone signalling. These findings reveal an unanticipated mechanism through which a transcriptional repressor of hormone signalling can directly recognize DNA and regulate transcription in higher plants.


Assuntos
Arabidopsis/genética , Arabidopsis/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Compostos Heterocíclicos com 3 Anéis/metabolismo , Lactonas/metabolismo , Reguladores de Crescimento de Planta/metabolismo , Transdução de Sinais/genética , Transcrição Genética , Antocianinas/biossíntese , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Genes de Plantas/genética , Reguladores de Crescimento de Planta/biossíntese , Folhas de Planta/anatomia & histologia , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Brotos de Planta/genética , Brotos de Planta/crescimento & desenvolvimento , Brotos de Planta/metabolismo , Regiões Promotoras Genéticas , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
4.
Gene ; 752: 144788, 2020 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-32439375

RESUMO

Primulina genus is an ideal wild ornamental flower and emerging model for studying biosynthesis, diversity, and evolution of flower pigment. However, the molecular mechanism underlying anthocyanin biosynthesis and regulation in Primulina remains unknown. Here, changes in anthocyanin content and the expression profiles of anthocyanin biosynthetic structural genes were examined in developing Primulina swinglei flowers and three other organs. Seventy-three R2R3-MYB transcription factor genes were identified from transcriptome of P. swinglei flowers, two of which, PsMYB1 and PsMYB2, are candidate regulators of anthocyanin biosynthesis according to clustering analysis. Furthermore, transient over-expression studies using tobacco leaves showed distinct pigment accumulation following co-infection with PsMYB1 and MrbHLH1 (a previously confirmed anthocyanin regulator from Morella rubra). Additionally, dual luciferase assays showed that PsMYB1 trans-activated the PsANS promoter, with the addition of MrbHLH1 resulting in a 5-fold increase in the intensity of this interaction. PsMYB1 did not, however, have any effect on the PsF3H promoter. The expression profile and dual luciferase assays showed that PsMYB2 plays no roles in anthocyanin regulation. Therefore, PsMYB1 is proposed to be the transcription factor gene regulating anthocyanin biosynthesis in P. swinglei.


Assuntos
Antocianinas/biossíntese , Antocianinas/genética , Fatores de Transcrição/genética , Sequência de Aminoácidos/genética , Antocianinas/metabolismo , Proteínas de Arabidopsis/metabolismo , Flores/genética , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica de Plantas/genética , Genes de Plantas/genética , Lamiales/genética , Magnoliopsida/genética , Pigmentação/genética , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética , Tabaco/genética , Transativadores/genética , Fatores de Transcrição/metabolismo , Transcriptoma/genética
5.
PLoS One ; 15(5): e0232090, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32357153

RESUMO

Dihydroflavonol 4-reductase (DFR), a key enzyme involved in the biosynthesis of anthocyanins, has been cloned from various species. However, little research has been conducted on this enzyme in ferns, which occupy a unique evolutionary position. In this study, we isolated two novel DFR genes from the fern Dryopteris erythrosora. In vitro enzymatic analysis revealed that DeDFR1 and DeDFR2 enzymes can catalyze dihydrokaempferol and dihydroquercetin but cannot catalyze dihydromyricetin. Amino acid sequence analysis showed that DeDFR1 and DeDFR2 have an arginine at the same substrate-specificity-determining site as that in the ferns Salvinia cucullata and Azolla filiculoides. Thus, we speculate that the Arg-type DFR is a new DFR functional type. To further verify the substrate preferences of the Arg-type DFR, an amino acid substitution assay was conducted. When N133 was mutated to R133, Arabidopsis DFR protein completely lost its catalytic activity for dihydromyricetin, as observed for DeDFR1 and DeDFR2. Additionally, heterologous expression of DeDFR2 in the Arabidopsis tt3-1 mutant resulted in increasing anthocyanin accumulation. In summary, DeDFR1 and DeDFR2 are considered to be a new type of DFR with unique structures and functions. The discovery of the Arg-type DFR provides new insights into the anthocyanin biosynthesis pathway in ferns.


Assuntos
Oxirredutases do Álcool/genética , Antocianinas/biossíntese , Dryopteris/genética , Oxirredutases do Álcool/metabolismo , Sequência de Aminoácidos , Catálise , Dryopteris/enzimologia , Dryopteris/metabolismo , Mutagênese Sítio-Dirigida , Filogenia , Alinhamento de Sequência , Especificidade por Substrato
6.
PLoS One ; 15(5): e0232986, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32407419

RESUMO

Here we focus on the highly conserved MYB-bHLH-WD repeat (MBW) transcriptional complex model in eggplant, which is pivotal in the transcriptional regulation of the anthocyanin biosynthetic pathway. Through a genome-wide approach performed on the recently released Eggplant Genome (cv. 67/3) previously identified, and reconfirmed by us, members belonging to the MBW complex (SmelANT1, SmelAN2, SmelJAF13, SmelAN1) were functionally characterized. Furthermore, a regulatory R3 MYB type repressor (SmelMYBL1), never reported before, was identified and characterized as well. Through a qPCR approach, we revealed specific transcriptional patterns of candidate genes in different plant tissue/organs at two stages of fruit development. Two strategies were adopted for investigating the interactions of bHLH partners (SmelAN1, SmelJAF13) with MYB counterparts (SmelANT1, SmelAN2 and SmelMYBL1): Yeast Two Hybrid (Y2H) and Bimolecular Fluorescent Complementation (BiFC) in A. thaliana mesophylls protoplast. Agro-infiltration experiments highlighted that N. benthamiana leaves transiently expressing SmelANT1 and SmelAN2 showed an anthocyanin-pigmented phenotype, while their co-expression with SmelMYBL1 prevented anthocyanin accumulation. Our results suggest that SmelMYBL1 may inhibits the MBW complex via the competition with MYB activators for bHLH binding site, although this hypothesis requires further elucidation.


Assuntos
Antocianinas/biossíntese , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo , Solanum melongena/genética , Solanum melongena/metabolismo , Sequência de Aminoácidos , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Genes Reguladores , Família Multigênica , Filogenia , Plantas Geneticamente Modificadas , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Tabaco/genética , Tabaco/metabolismo
7.
PLoS One ; 15(4): e0231729, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32330148

RESUMO

Radish (Raphanus sativus L.), is an important root vegetable crop grown worldwide, and it contains phyto-anthocyanins. However, only limited studies have been conducted to elucidate the molecular mechanisms underlying anthocyanin biosynthesis in the different color variants of the radish fleshy root. In this study, Illumina paired-end RNA-sequencing was employed to characterize the transcriptomic changes in seven different types of radish fleshy roots. Approximately, 126 co-modulated differentially expressed genes were obtained, and most DEGs were more likely to participate in anthocyanin biosynthesis, including two transcription factors RsMYB_9 and RsERF070, and four functional genes RsBRICK1, RsBRI1-like2, RsCOX1, and RsCRK10. In addition, some related genes such as RsCHS, RsCHI, RsANS, RsMT2-4, RsUF3GT, glutathione S-transferase F12, RsUFGT78D2-like and RsUDGT-75C1-like significantly contributed to the regulatory mechanism of anthocyanin biosynthesis in the radish cultivars. Furthermore, gene ontology analysis revealed that the anthocyanin-containing compound biosynthetic process, anthocyanin-containing compound metabolic process, and significantly enriched pathways of the co-modulated DEGs were overrepresented in these cultivars. These results will expand our understanding of the complex molecular mechanism underlying anthocyanin synthesis-related genes in radish.


Assuntos
Antocianinas/biossíntese , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Raphanus/genética , Verduras/genética , Vias Biossintéticas/genética , Ontologia Genética , Genes de Plantas/genética , Sequenciamento de Nucleotídeos em Larga Escala , Proteínas de Plantas/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , RNA-Seq , Raphanus/metabolismo , Verduras/metabolismo
8.
Plant Mol Biol ; 103(4-5): 443-456, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32270430

RESUMO

KEY MESSAGE: The simultaneous expression of AmRosea1 and AmDelila transcription factors from snapdragon can activate the anthocyanin pathway in orange carrots, leading to the synthesis and accumulation of anthocyanins in the taproots. Anthocyanins are phenolic compounds produced in various parts of plants. They are used as natural food dyes and are reported as beneficial antioxidants for humans. Black carrot is an important source for anthocyanins; however, the reason for the lack of anthocyanin production in the orange carrot is unknown. Anthocyanins are synthesized by a specific branch of the phenylpropanoid pathway that has previously been reported to be activated by a triad of R2R3-MYB, basic helix-loop helix (bHLH) and WD40 transcription factors (TFs). In the current study, orange carrots were turned purple by simultaneous expression of R2R3-MYB and bHLH TFs, i.e. AmRosea1 and AmDelila from snapdragon (Antirrhinum majus). Simultaneous transgenic expression of the TFs under a constitutive promoter in the orange carrot cultivar 'Danvers 126' lead to consistent upregulation of anthocyanin-related biosynthetic genes and significant accumulation of anthocyanins in leaves, stems and taproots. Highest overall content of soluble anthocyanins in the taproot among the transformants amounted to 44.38 mg g-1 dry weight. The anthocyanin profile of the transformants were significantly different from the profile in the reference black carrot 'Deep Purple'. The main anthocyanins present in the transformed taproots were cyanidin 3-xylosyl(sinapoylglucosyl)galactoside, whereas the main anthocyanin present in Deep Purple was cyanidin 3-xylosyl(feruloylglucosyl)galactoside. This study confirms the presence of the necessary biosynthetic genes in orange carrots for production of anthocyanins and demonstrates the absence of suitable R2R3-MYB and bHLH TFs for stimulating anthocyanin biosynthesis in the orange carrot.


Assuntos
Antocianinas/biossíntese , Antocianinas/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Daucus carota/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Vias Biossintéticas/genética , Cor , Daucus carota/genética , Genes de Plantas/genética , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Fenótipo , Pigmentos Biológicos/biossíntese , Pigmentos Biológicos/genética , Folhas de Planta/metabolismo , Plantas Geneticamente Modificadas , Fatores de Transcrição , Transformação Genética
9.
Artigo em Inglês | MEDLINE | ID: mdl-32247249

RESUMO

Anthocyanins in cornflower (Centaurea cyanus) is catalysed by a set of biosynthesis genes, however, the potential mechanism of transcriptional regulation remains unclear. In the present study, we traced the dynamic changes of petal colour development from white to violet and finally to blue on the same petal in cornflower. Pigment analysis showed that anthocyanin accumulation dramatically increased with petal colour development. Subsequently, nine libraries from above three colour regions were constructed for RNA-seq and 105,506 unigenes were obtained by de novo assembling. The differentially expressed genes among three colour regions were significantly enriched in the phenylpropanoid biosynthesis and flavonoid biosynthesis pathways, leading to the excavation and analysis of 46 biosynthesis genes involved in this process. Furthermore, four R2R3-CcMYBs clustered into subgroup 4 or subgroup 6 and one CcbHLH1 clustered into IIIf subgroup were screened out by phylogenetic analysis with Arabidopsis homologues. The promoters of flavanone 3-hydroxylase (CcF3H) and dihydroflavonol 4-reductase (CcDFR) were further isolated to investigate upstream regulation mechanism. CcMYB6-1 significantly upregulated the activity of above two promoters and stimulated anthocyanin accumulation by dual luciferase assay and transient expression in tobacco leaves, and its activity was obviously enhanced when co-infiltrated with CcbHLH1. Moreover, both yeast two-hybrid and bimolecular fluorescence complementation assays indicated the protein-protein interaction between these two activators. Based on these obtained results, it reveals that CcMYB6-1 and CcbHLH1 are two novel transcription factors synergistically involved in regulating anthocyanin biosynthesis. This study provides insights into the regulatory mechanism of anthocyanin accumulation in cornflower.


Assuntos
Antocianinas , Centaurea , Regulação da Expressão Gênica de Plantas , Fatores de Transcrição , Antocianinas/biossíntese , Antocianinas/genética , Centaurea/classificação , Centaurea/genética , Centaurea/metabolismo , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Fatores de Transcrição/metabolismo
10.
Artigo em Inglês | MEDLINE | ID: mdl-32005382

RESUMO

Ultraviolet-B (UV-B) radiation promotes anthocyanin synthesis in many plants. Although several transcription factors promote anthocyanin synthesis in response to UV-B radiation, the underlying mechanism remains unclear. In this study, the MdWRKY72 transcription factor gene was isolated from the 'Taishanzaoxia' apple genome. Quantitative real-time PCR analyses revealed that the genes encoding enzymes and transcription factors involved in the anthocyanin synthesis pathway (MdANS, MdDFR, MdUFGT, and MdMYB1) were more highly expressed in MdWRKY72-overexpressing transgenic calli than in the wild-type 'Orin' apple calli. The results indicated that MdWRKY72 increases anthocyanin synthesis in transgenic calli exposed to UV-B radiation. The results of a gel shift assay and chromatin immunoprecipitation proved that MdWRKY72 promotes MdMYB1 expression indirectly by binding to a W-box element in the MdHY5 promoter and directly by binding to a W-box element in the MdMYB1 promoter. Thus, MdWRKY72 increases anthocyanin synthesis via direct and indirect mechanisms. These findings may be useful for elucidating the molecular mechanism underlying UV-B-induced anthocyanin synthesis mediated by MdWRKY72.


Assuntos
Antocianinas/biossíntese , Malus/genética , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Sequência de Aminoácidos , Antocianinas/genética , Sequência de Bases , Imunoprecipitação da Cromatina , Ensaio de Desvio de Mobilidade Eletroforética , Malus/metabolismo , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Alinhamento de Sequência , Fatores de Transcrição/química , Fatores de Transcrição/metabolismo
11.
Artigo em Inglês | MEDLINE | ID: mdl-32005403

RESUMO

Sumoylation is one of post-translational modification (PTM) in which SUMO (small ubiquitin-like modifier) are covalently conjugated to protein substrates through a range of biochemical steps. This paper presents evidence that SUMO E3 ligase SIZ1 positively regulates anthocyanin accumulation. Loss-of-function siz1 mutant seedlings exhibit anthocyanin accumulation-reduced phenotype under high light conditions. Moreover, SIZ1 interacts and sumoylates MYB75/PAP1, a key transcription factor in anthocyanin accumulation. Loss-of-function siz1 or K246R substitution in MYB75 blocked SIZ1-mediated sumoylation in vitro and in vivo. Anthocyanin accumulation in mutant myb75-c can not be rescued by expressing MYB75K246R, but expression of wild-type MYB75WT complements the mutant phenotype. It suggested that sumoylation is important for MYB75 function. We further prove that sumoylation is essential for MYB75 protein stability. And SIZ1 is involved in the light-induced accumulation of anthocyanins. Our findings reveal an important role for sumoylation of MYB in regulation of anthocyanin accumulation in plants.


Assuntos
Antocianinas/biossíntese , Proteínas de Arabidopsis/genética , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Ligases/genética , Fatores de Transcrição/genética , Sequência de Aminoácidos , Antocianinas/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/metabolismo , Ligases/metabolismo , Luz , Plântula/genética , Plântula/metabolismo , Sumoilação , Fatores de Transcrição/química , Fatores de Transcrição/metabolismo
12.
Gene ; 743: 144484, 2020 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-32081694

RESUMO

Glutathione S-transferases (GSTs) are a large complex family of enzymes (EC 2.5.1.18) that play vital roles in flavonoid metabolism and plant growth and development and are responsive to heavy metal stress. However, knowledge about GST genes in radish (a vegetable crop with an extraordinary capacity to adapt to heavy metal stresses) is limited. Therefore, it is critical to identify putative candidate GST genes responsible for heavy metal stress tolerance and anthocyanin biosynthesis. In this study, we first identified 82 R. sativus GST (RsGST) genes using various bioinformatic approaches, and their expression profiles were characterized from RNAseq data. These RsGST genes could be grouped into 7 major subclasses: tau (43 members), phi (21 members), tetrachlorohydroquinone dehalogenase (7 members), dehydroascorbat reductase (5 members), zeta (3 members), lambda (2 members) and theta (1 member). In addition, most of the RsGST genes showed organ-specific expression in our study. Moreover, the transcripts of RsGSTF12-1 and RsGSTF12-2, belonging to the phi class, might be candidates encoding anthocyanin transporters in carmine radish, whereas the tau class, consisting of RsGSTU13-1, RsGSTU19, RsGSTU24-1, and RsGSTU3, and theta class, consisting of RsGSTT1-1, might be defend radish against adverse heavy metal stresses. These results will aid in understanding the functions of the GST family related to heavy metal stress and anthocyanin biosynthesis, thereby potentially improving radish breeding programs for high-pigment-content material as well as HM-tolerant material.


Assuntos
Antocianinas/biossíntese , Glutationa Transferase/genética , Metais Pesados/efeitos adversos , Proteínas de Plantas/genética , Raphanus/enzimologia , Adaptação Fisiológica , Biologia Computacional , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genes de Plantas/genética , Glutationa Transferase/isolamento & purificação , Glutationa Transferase/metabolismo , Filogenia , Melhoramento Vegetal , Proteínas de Plantas/isolamento & purificação , Proteínas de Plantas/metabolismo , Raízes de Plantas , RNA-Seq , Raphanus/genética , Raphanus/metabolismo , Estresse Fisiológico/efeitos dos fármacos
13.
Planta ; 251(3): 60, 2020 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-32030477

RESUMO

MAIN CONCLUSION: The phosphorylation status of MYB75 at T-131 affects protein stability, flavonoid profiles, and patterns of gene expression. The Arabidopsis transcription factor Myeloblastosis protein 75 (MYB75, AT1G56650) is known to act as a positive transcriptional regulator of genes required for flavonoid and anthocyanin biosynthesis. MYB75 was also shown to negatively regulate lignin and other secondary cell wall biosynthetic genes (Bhargava et al. in Plant Physiol 154(3):1428-1438, 2010). While transcriptional regulation of MYB75 has been described in numerous publications, little is known about post-translational control of MYB75 protein function. In a recent publication, light-induced activation of a MAP kinase (MPK4, AT4G01370) in Arabidopsis was reported to lead to MYB75 phosphorylation at two canonical MPK target sites, threonines, T-126 and T-131. This double phosphorylation event positively influenced MYB75 protein stability (Li et al. in Plant Cell 28(11):2866-2883, 2016). We have examined this phenomenon through use of phosphomutant forms of MYB75 and found that MYB75 is phosphorylated primarily at T-131, and that the phosphorylation of MYB75 recombinant protein in vitro can be catalyzed by multiple MAP kinases, including MPK3 (AT3G45640), MPK6 (AT2G43790), MPK4 and MPK11 (AT1G01560). We also demonstrate that MYB75 can bind to a large number of Arabidopsis MPK's in vitro, suggesting it could be a target of multiple signalling pathways. The impact of MYB75 phosphorylation at T-131 on the function of this transcription factor, in terms of localization, stability, and protein-protein interactions with known binding partners was examined in transgenic lines expressing phosphomimic and phosphonull versions of MYB75, to capture the behaviour of permanently phosphorylated and unphosphorylated MYB75 protein, respectively. In addition, we describe how ectopic over-expression of different phosphovariant forms of MYB75 (MYB75WT, MYB75T131A, and MYB75T131E) affects flavonoid biochemical profiles and global changes of gene expression in the corresponding transgenic Arabidopsis plants.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Fatores de Transcrição/metabolismo , Antocianinas/biossíntese , Antocianinas/química , Arabidopsis/efeitos dos fármacos , Arabidopsis/genética , Arabidopsis/efeitos da radiação , Proteínas de Arabidopsis/genética , Vias Biossintéticas/genética , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Genes de Plantas , Luz , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Fosforilação/efeitos dos fármacos , Fosforilação/efeitos da radiação , Plantas Geneticamente Modificadas , Ligação Proteica/efeitos dos fármacos , Ligação Proteica/efeitos da radiação , Estabilidade Proteica/efeitos dos fármacos , Transporte Proteico , Proteínas Recombinantes/metabolismo , Saccharomyces cerevisiae/metabolismo , Plântula/efeitos dos fármacos , Plântula/metabolismo , Plântula/efeitos da radiação , Sacarose/farmacologia , Fatores de Transcrição/genética
14.
Planta ; 251(3): 61, 2020 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-32036464

RESUMO

MAIN CONCLUSION: MiMYB1 and MibHLH2 play key roles in anthocyanin biosynthesis in Matthiola incana flowers. We established a transient expression system using Turnip mosaic virus vector in M. incana. Garden stock (Matthiola incana (L.) R. Br.) is a popular flowering plant observed from winter to spring in Japan. Here we observed that anthocyanin accumulation in 'Vintage Lavender' increased with flower development, whereas flavonol accumulation remained constant throughout flower development. We obtained five transcription factor genes, MiMYB1, MibHLH1, MibHLH2, MiWDR1, and MiWDR2, from M. incana floral cDNA contigs. Yeast two-hybrid analyses revealed that MiMYB1 interacted with MibHLH1, MibHLH2, and MiWDR1, but MiWDR2 did not interact with any transcription factor. Expression levels of MiMYB1 and MibHLH2 increased in petals during floral bud development. Their expression profiles correlated well with the temporal profiles of MiF3'H, MiDFR, MiANS, and Mi3GT transcripts and anthocyanin accumulation profile. On the other hand, MibHLH1 was expressed weakly in all organs of 'Vintage Lavender'. However, high expression levels of MibHLH1 were detected in petals of other cultivars with higher levels of anthocyanin accumulation than 'Vintage Lavender'. MiWDR1 and MiWDR2 maintained constant expression levels in petals during flower development and vegetative organs. Transient MiMYB1 expression in 1-month-old M. incana seedlings using a Turnip mosaic virus vector activated transcription of the endogenous anthocyanin biosynthetic genes MiF3'H, MiDFR, and MiANS and induced ectopic anthocyanin accumulation in leaves. Therefore, MiMYB1 possibly interacts with MibHLH2 and MiWDR1, and this trimeric protein complex activates the transcription of anthocyanin biosynthetic genes in M. incana flowers. Moreover, MibHLH1 acts as an enhancer of anthocyanin biosynthesis with the MiMYB1-MibHLH2-MiWDR1 complex. This study revealed the molecular mechanism involved in the regulation of anthocyanin accumulation levels in M. incana flowers.


Assuntos
Antocianinas/metabolismo , Brassicaceae/genética , Flores/genética , Genes de Plantas , Pigmentação/genética , Antocianinas/biossíntese , Vias Biossintéticas/genética , Flavonoides/metabolismo , Regulação da Expressão Gênica de Plantas , Potyvirus/fisiologia , Ligação Proteica , Plântula/virologia , Fatores de Tempo , Tabaco/virologia
15.
Genes (Basel) ; 11(1)2020 01 10.
Artigo em Inglês | MEDLINE | ID: mdl-31936856

RESUMO

To elucidate the effect of low temperature on anthocyanin biosynthesis in purple head Chinese cabbage, we analyzed anthocyanin accumulation and related gene expression in the seedlings of purple head Chinese cabbage, white head parent Chinese cabbage, and its purple male parent under a normal 25 °C temperature and a low 12 °C temperature. Anthocyanin accumulation in purple lines was strongly induced by low temperature, and the total anthocyanin content of seedlings was significantly enhanced. In addition, nearly all phenylpropanoid metabolic pathway genes (PMPGs) were down-regulated, some early biosynthesis genes (EBGs) were up-regulated, and nearly all late biosynthesis genes (LBGs) directly involved in anthocyanin biosynthesis showed higher expression levels in purple lines after low-temperature induction. Interestingly, a R2R3-MYB transcription factor (TF) gene 'BrMYB2' and a basic-helix-loop-helix (bHLH) regulatory gene 'BrTT8' were highly up-regulated in purple lines after low temperature induction, and two negative regulatory genes 'BrMYBL2.1' and 'BrLBD38.2' were up-regulated in the white line. BrMYB2 and BrTT8 may play important roles in co-activating the anthocyanin structural genes in purple head Chinese cabbage after low-temperature induction, whereas down-regulation of BrMYB2 and up-regulation of some negative regulators might be responsible for white head phenotype formation. Data presented here provide new understanding into the anthocyanin biosynthesis mechanism during low temperature exposure in Brassica crops.


Assuntos
Antocianinas/biossíntese , Antocianinas/genética , Brassica rapa/genética , Brassica/genética , Temperatura Baixa , Criobiologia/métodos , Expressão Gênica/genética , Regulação da Expressão Gênica de Plantas/genética , Proteínas de Plantas/genética , Plântula/genética , Temperatura , Fatores de Transcrição/genética
16.
J Photochem Photobiol B ; 203: 111779, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31927487

RESUMO

Light is a key environmental factors affecting anthocyanin accumulation in plants. Ubiquitin E3 ligase COP1 has been proved to be a negative regulator involved in light-regulated plant development process, whereas the function and expression specificity of COP1 in anthocyanin biosynthesis in sweet cherry remains unclear. In the present study, we identified a COP1 in sweet cherry, named PacCOP1, it exhibited apparent different expression patterns in red-colored 'Hongdeng' and bi-colored 'Satonishiki', with increasing trend largely in 'Satonishiki', but decreasing trend in 'Hongdeng' after veraison, which was contrary to their variation tendency of anthocyanin content. While the expression abundance of anthocyanin biosynthesis related genes were largely increased after veraison, in accordance with anthocyanin content. Correlation analysis proved that the expression of PacCOP1 was negative correlated with the major genes on anthocyanin accumulation in 'Hongdeng' and 'Satonishiki' fruit, in especial PacDFR, PacANS, PacMYBA and PacbHLH33. Furthermore, over-expression of PacCOP1 in Arabidopsis displayed increased COP1 transcript level with negligible pigmentation and corresponding lower expression level of AtPAP1, AtDFR, AtLDOX, and AtUFGT. These results revealed the negative regular role of PacCOP1 in anthocyanin biosynthesis by repressing the PacMYBA transcription level, followed by down-regulating the structural genes expression abundance, eventually leading to attenuated anthocyanin accumulation in fruits.


Assuntos
Antocianinas/biossíntese , Proteínas de Plantas/metabolismo , Prunus avium/enzimologia , Ubiquitina-Proteína Ligases/metabolismo , Sequência de Aminoácidos , Arabidopsis/metabolismo , Frutas/metabolismo , Regulação da Expressão Gênica de Plantas , Filogenia , Pigmentação/genética , Proteínas de Plantas/classificação , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/metabolismo , Prunus avium/metabolismo , Alinhamento de Sequência , Ubiquitina-Proteína Ligases/classificação , Ubiquitina-Proteína Ligases/genética
17.
Food Chem ; 314: 126170, 2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-31978717

RESUMO

Regulated deficit irrigation (RDI) is a new type of water-saving irrigation technology developed in recent years which was well suited to arid and semi-arid grape plant areas. The anthocyanin synthesis of grapes under RDI was revealed through omics in this study. RDI slightly decreased the hundred-grain weight and increased the soluble solid content, juice pH, reducing sugar content, and total anthocyanin content. Meanwhile, the total acid content decreased before ripening. Transcriptomics and metabolomics analyses revealed that large numbers of differentially expressed genes (DEGs) and significantly changed metabolites (SCMs) were filtered in the RDI groups. RDI1 with 30% ETc upregulated 7 related gene expression levels in the anthocyanin biosynthetic pathway and also increased some metabolites contents. Eventually, the contents of most monomeric anthocyanins in the RDI groups were increased, and the proportion of Mv increased in the ripe grapes of the RDI groups. In all, RDI is a useful water-saving irrigation method which could also increase anthocyanin content in grapes.


Assuntos
Antocianinas/análise , Metabolômica , Transcriptoma , Vitis/química , Antocianinas/biossíntese , Frutas/química
18.
Food Chem ; 310: 125862, 2020 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-31767480

RESUMO

'Friar' plum (Prunus salicina Lindl.) fruit were transferred to shelf life (25 °C) following different storage periods at low (0 °C) or intermediate (5 °C) temperature. The earliest flesh reddening appeared in plums during shelf life removed after 28 d at 0 °C and 14 d at 5 °C, respectively, in comparison with turning yellow in plums remained at 25 °C immediately after harvest. The flesh reddening developed rapidly thereafter. Anthocyanins, in particular, cyanidin 3-O-glucoside, significantly accumulated in the reddening tissue, and activities of enzymes associated with the phenylpropanoid pathway were considerably activated after the removal. The removal elicited extremely high ethylene production in plums, which might mediate the activation of the anthocyanin biosynthesis in response to cold stress signal. The results provided a potential approach for postharvest regulation of flesh color and accumulation of bioactive substances in plums, which could lead to attractive attributes and health-promoting effects on consumers.


Assuntos
Antocianinas/biossíntese , Temperatura Baixa , Cor , Prunus/metabolismo , Antocianinas/isolamento & purificação , Antocianinas/metabolismo , Antioxidantes/metabolismo , Etilenos , Frutas/metabolismo , Prunus domestica , Temperatura
19.
Biosci Biotechnol Biochem ; 84(4): 797-799, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31790630

RESUMO

Insect gall structures have many characteristic forms and colors, which are distinguishable from host plants. In this study, we identified an anthocyanin from red color insect galls and revealed that the anthocyanin biosynthesis of plants was induced by the gall extracts. The galling insects presumably regulate the anthocyanin biosynthesis of host plants to protect their larvae from environmental stresses.


Assuntos
Antocianinas/química , Ceratopogonidae/fisiologia , Fagus/parasitologia , Galactosídeos/química , Interações Hospedeiro-Parasita , Animais , Antocianinas/biossíntese , Ceratopogonidae/crescimento & desenvolvimento , Fagus/metabolismo , Larva/fisiologia
20.
J Sci Food Agric ; 100(4): 1505-1514, 2020 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-31756273

RESUMO

BACKGROUND: Greater proportions of purple tea buds and leaves usually appear in the summer, which seriously affects the color and taste quality of green tea products, yet the metabolism of purple tea shoots in summer remains unclear. Here, the metabolomic profiles and gene expression of related flavonoid metabolic pathways in the purple and normal green shoots of 'Longjing 43', and the quality of green tea made with these two phenotypes, were analyzed and compared. RESULTS: Differential metabolites identified using high-performance liquid chromatography-Orbitrap/mass spectrometry indicated that anthocyanin biosynthesis in purple leaves was enriched, with higher levels of anthocyanidins (delphinidin-hexose-coumaroyl showed the greatest increase), proanthocyanidins (oligomers of catechins) and kaempferol glycoside. Expression patterns of the genes ANR, ANS, FLS, LAR, C4H, PAL, CHI, CHS and DFR revealed that the metabolism of anthocyanin is positively regulated by high temperature and/or light levels in summer. Gas chromatography-mass spectrometry results showed that, in purple tea shoots, the metabolism of carbohydrates was enriched whereas that of amino acids was diminished, while their mannose, fructose, d-galactose, sorbose and d-glucose contents were more than double those found in green leaves. A sensory evaluation confirmed that a greater quantity of purple shoots had a greater negative impact on green tea quality because of a bitter taste and dark color (leaves and infusions were tested). CONCLUSIONS: These results highlight the need for and possibility of improving commercial tea quality via cultivation that controls the temperature or light of tea gardens during the summer. © 2019 Society of Chemical Industry.


Assuntos
Antocianinas/biossíntese , Camellia sinensis/metabolismo , Brotos de Planta/metabolismo , Camellia sinensis/química , Camellia sinensis/genética , Camellia sinensis/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Folhas de Planta/química , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Brotos de Planta/química , Brotos de Planta/genética , Brotos de Planta/crescimento & desenvolvimento , Estações do Ano
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