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1.
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
2.
J Agric Food Chem ; 67(32): 8783-8793, 2019 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-31310107

RESUMO

Red-fleshed apples are popular as a result of their high anthocyanin content. MdMYB10 and its homologues are known to be important regulators of anthocyanin synthesis in apple, but the roles of other transcription factors are not well-understood. Here, we explored the role of MdWRKY11 in regulating anthocyanin synthesis in apple flesh. Overexpression of MdWRKY11 in apple callus could significantly promote anthocyanin accumulation, and the expression of some MYB transcription factors and structural genes increased significantly. In binding analyses, MdWRKY11 bound to W-box cis-elements in the promoters of MdMYB10, MdMYB11, and MdUFGT. However, MdWRKY11 did not interact with MdMYB10, MdbHLH3, or MdWD40 proteins, the members of the MBW complex. Sequence analyses revealed that another W-box cis-element was present in the promoter of MdHY5 (encoding a photoresponse factor), and MdWRKY11 was able to bind to the promoter of MdHY5 and promote its activity. Our findings clarify the role of MdWRKY11 in anthocyanin synthesis in red-fleshed apple and imply that other novel genes may be involved in anthocyanin synthesis.


Assuntos
Antocianinas/biossíntese , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Regulação da Expressão Gênica de Plantas , Malus/metabolismo , Proteínas de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Fatores de Transcrição de Zíper de Leucina Básica/genética , Frutas/genética , Frutas/crescimento & desenvolvimento , Frutas/metabolismo , Malus/genética , Malus/crescimento & desenvolvimento , Proteínas de Plantas/genética , Fatores de Transcrição/genética
3.
BMC Plant Biol ; 19(1): 287, 2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-31262258

RESUMO

BACKGROUND: The majority of apricot (Prunus armeniaca L.) cultivars display orange or yellow background skin, whereas some cultivars are particularly preferred by consumers because of their red blushed skin on the background. RESULTS: In this study, two blushed ('Jianali' and 'Hongyu') and two nonblushed ('Baixing' and 'Luntaixiaobaixing') cultivars were used to investigate the formation mechanism of blushed skin in apricots. High-performance liquid chromatography (HPLC) analysis showed that the blushed cultivars accumulated higher cyanidin-3-O-glucoside, cyanidin-3-O-rutinoside and peonidin-3-O-rutinoside levels during fruit ripening than the nonblushed cultivars. Based on coexpression network analysis (WGCNA), a putative anthocyanin-related R2R3-MYB, PaMYB10, and seven structural genes were identified from transcriptome data. The phylogenetic analysis indicated that PaMYB10 clustered in the anthocyanin-related MYB clade. Sequence alignments revealed that PaMYB10 contained a bHLH-interaction motif ([DE]Lx2[RK]x3Lx6Lx3R) and an ANDV motif. Subcellular localization analysis showed that PaMYB10 was a nuclear protein. Real-time qRT-PCR analysis demonstrated that the transcript levels of PaMYB10 and seven genes responsible for anthocyanin synthesis were significantly higher in blushed than in nonblushed apricots, which was consistent with the accumulation of anthocyanin. In addition, bagging significantly inhibited the transcript levels of PaMYB10 and the structural genes in 'Jianali' and blocked the red coloration and anthocyanin accumulation. Transient PaMYB10 overexpression in 'Luntaixiaobaixing' fruits resulted in the red blushed skin at the maturation stage. CONCLUSIONS: Taken together, these data reveal that three anthocyanins are responsible for the blushed skin of apricots, identify PaMYB10 as a positive regulator of anthocyanin biosynthesis in apricots, and demonstrate that blush formation depends on light.


Assuntos
Antocianinas/biossíntese , Regulação da Expressão Gênica de Plantas , Pigmentos Biológicos/biossíntese , Proteínas de Plantas/genética , Prunus armeniaca/fisiologia , Fatores de Transcrição/genética , Sequência de Aminoácidos , Antocianinas/genética , Cromatografia Líquida de Alta Pressão , Cor , Frutas/genética , Frutas/fisiologia , Glucosídeos/biossíntese , Glucosídeos/genética , Filogenia , Pigmentos Biológicos/genética , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Prunus armeniaca/genética , Alinhamento de Sequência , Fatores de Transcrição/química , Fatores de Transcrição/metabolismo
4.
BMC Plant Biol ; 19(1): 317, 2019 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-31307384

RESUMO

BACKGROUND: Anthocyanins, which are colored pigments, have long been used as food and pharmaceutical ingredients due to their potential health benefits, but the intermediate signals through which environmental or developmental cues regulate anthocyanin biosynthesis remains poorly understood. Fleshy fruits have become a good system for studying the regulation of anthocyanin biosynthesis, and exploring the mechanism underlying pigment metabolism is valuable for controlling fruit ripening. RESULTS: The present study revealed that ABA accumulated during Lycium fruit ripening, and this accumulation was positively correlated with the anthocyanin contents and the LbNCED1 transcript levels. The application of exogenous ABA and of the ABA biosynthesis inhibitor fluridon increased and decreased the content of anthocyanins in Lycium fruit, respectively. This is the first report to show that ABA promotes the accumulation of anthocyanins in Lycium fruits. The variations in the anthocyanin content were consistent with the variations in the expression of the genes encoding the MYB-bHLH-WD40 transcription factor complex or anthocyanin biosynthesis-related enzymes. Virus-induced LbNCED1 gene silencing significantly slowed fruit coloration and decreased both anthocyanin and ABA accumulation during Lycium fruit ripening. An qRT-PCR analysis showed that LbNCED1 gene silencing clearly reduced the transcript levels of both structural and regulatory genes in the flavonoid biosynthetic pathway. CONCLUSIONS: Based on the results, a model of ABA-mediated development-dependent anthocyanin biosynthesis and fruit coloration during Lycium fruit maturation was proposed. In this model, the developmental cues transcriptionally activates LbNCED1 and thus enhances accumulation of the phytohormone ABA, and the accumulated ABA stimulates transcription of the MYB-bHLH-WD40 transcription factor complex to upregulate the expression of structural genes in the flavonoid biosynthetic pathway and thereby promoting anthocyanin production and fruit coloration. Our results provide a valuable strategy that could be used in practice to regulate the ripening and quality of fresh fruit in medicinal and edible plants by modifying the phytohormone ABA.


Assuntos
Ácido Abscísico/metabolismo , Antocianinas/biossíntese , Frutas/metabolismo , Lycium/metabolismo , Pigmentação , Reguladores de Crescimento de Planta/metabolismo , Dioxigenases/genética , Frutas/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Inativação Gênica , Genes de Plantas , Lycium/genética , Lycium/crescimento & desenvolvimento , Proteínas de Plantas/genética , Transdução de Sinais
5.
BMC Plant Biol ; 19(1): 232, 2019 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-31159725

RESUMO

BACKGROUND: Compared with white-fleshed sweetpotato (WFSP), purple-fleshed sweetpotato (PFSP) is a desirable resource for functional food development because of the abundant anthocyanin accumulation in its tuberous roots. Some studies have shown that the expression regulation mediated by miRNA plays an important role in anthocyanin biosynthesis in plants. However, few miRNAs and their corresponding functions related to anthocyanin biosynthesis in tuberous roots of sweetpotato have been known. RESULTS: In this study, small RNA (sRNA) and degradome libraries from the tuberous roots of WFSP (Xushu-18) and PFSP (Xuzishu-3) were constructed, respectively. Totally, 191 known and 33 novel miRNAs were identified by sRNA sequencing, and 180 target genes cleaved by 115 known ib-miRNAs and 5 novel ib-miRNAs were identified by degradome sequencing. Of these, 121 miRNAs were differently expressed between Xushu-18 and Xuzishu-3. Integrated analysis of sRNA, degradome sequencing, GO, KEGG and qRT-PCR revealed that 26 differentially expressed miRNAs and 36 corresponding targets were potentially involved in the anthocyanin biosynthesis. Of which, an inverse correlation between the expression of ib-miR156 and its target ibSPL in WFSP and PFSP was revealed by both qRT-PCR and sRNA sequencing. Subsequently, ib-miR156 was over-expressed in Arabidopsis. Interestingly, the ib-miR156 over-expressing plants showed suppressed abundance of SPL and a purplish phenotype. Concomitantly, upregulated expression of four anthocyanin pathway genes was detected in transgenic Arabidopsis plants. Finally, a putative ib-miRNA-target model involved in anthocyanin biosynthesis in sweetpotato was proposed. CONCLUSIONS: The results represented a comprehensive expression profiling of miRNAs related to anthocyanin accumulation in sweetpotato and provided important clues for understanding the regulatory network of anthocyanin biosynthesis mediated by miRNA in tuberous crops.


Assuntos
Antocianinas/biossíntese , Regulação da Expressão Gênica de Plantas , Ipomoea batatas/genética , MicroRNAs/genética , RNA de Plantas/genética , Antocianinas/genética , Sequenciamento de Nucleotídeos em Larga Escala , Ipomoea batatas/metabolismo , MicroRNAs/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , RNA de Plantas/metabolismo , Análise de Sequência de RNA
6.
J Agric Food Chem ; 67(26): 7399-7409, 2019 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-31244203

RESUMO

Flavonol synthase (FLS) belongs to the 2-oxoglutarate-dependent dioxygenase (2-ODD) superfamily. We isolated OsFLS from the rice ( Oryza sativa) cultivar "Ilmi" OsFLS includes highly conserved 2-ODD-specific motifs and FLS-specific regions. Recombinant OsFLS exhibited both FLS and flavanone 3ß-hydroxylase (F3H) activities, converting dihydroflavonols into flavonols and flavanones into dihydroflavonols, respectively, and more efficiently used dihydrokaempferol than dihydroquercetin as a substrate. OsFLS was expressed in both nonpigmented and pigmented rice seeds and was developmentally regulated during seed maturation. Transgenic tobacco ( Nicotiana tabacum) plants expressing OsFLS produced pale pink or white flowers with significantly increased levels of kaempferol-3- O-rutinoside and dramatically reduced levels of anthocyanin in their petals. Additionally, pod size and weight were reduced compared to the wild type. Several early and late biosynthetic genes of flavonoid were downregulated in the transgenic flowers. We demonstrated that OsFLS is a bifunctional 2-ODD enzyme and functions in flavonol production in planta.


Assuntos
Dioxigenases/genética , Dioxigenases/metabolismo , Oryza/enzimologia , Oxirredutases/genética , Oxirredutases/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Antocianinas/biossíntese , Cor , Flavonóis/biossíntese , Flores/genética , Flores/metabolismo , Regulação da Expressão Gênica de Plantas , Ácidos Cetoglutáricos/metabolismo , Oryza/genética , Tabaco/genética , Tabaco/metabolismo
7.
Int J Mol Sci ; 20(9)2019 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-31064132

RESUMO

Flavonoids play multiple roles in plant coloration and stress resistance and are closely associated with human health. Flavonoids and non-flavonoids (such as phenolic acids) are produced via the phenylpropanoid-derived pathway. Anthocyanidin synthase (ANS) catalyzes the synthesis of anthocyanins from leucoanthocyanidin in the flavonoids branched pathway. In this study, SmANS from Salvia miltiorrhiza was cloned and mainly localized in the endoplasmic reticulum (ER), plastids, Golgi, plasma membrane, and nucleus of tobacco epidermal cells, and was most highly expressed in purple petals in S. miltiorrhiza, whereas it showed almost no expression in white petals, green calyxes, and pistils in S. miltiorrhiza Bge f. alba. Overexpressed SmANS enhanced anthocyanin accumulation but reduced salvianolic acid B (SAB) and rosmarinic acid (RA) biosynthesis in S. miltiorrhiza and S. miltiorrhiza Bge f. alba plantlets, meanwhile, it restored the purple-red phenotype in S. miltiorrhiza Bge f. alba. These changes were due to reallocation of the metabolic flow, which was influenced by the SmANS gene. These findings indicate that SmANS not only plays a key role in anthocyanin accumulation in S. miltiorrhiza, but also acts as a "switch" for the coloration of S. miltiorrhiza Bge f. alba. This study provides baseline information for further research on flavonoids metabolism and improvement of anthocyanin or phenolic acid production by genetic engineering.


Assuntos
Antocianinas/biossíntese , Hidroxibenzoatos/metabolismo , Oxigenases/genética , Proteínas de Plantas/genética , Salvia miltiorrhiza/genética , Oxigenases/metabolismo , Proteínas de Plantas/metabolismo , Salvia miltiorrhiza/metabolismo
8.
Plant Sci ; 283: 165-176, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31128686

RESUMO

The effect of temperature on the concentrations of anthocyanins and endogenous plant hormones [abscisic acid (ABA), auxin, and cytokinin] were investigated using the detached berries of two related red-skinned cultivars cv. 'Aki Queen' and 'Ruby Roman' of the table grape Vitis labrusca L. × Vitis vinifera L. The total anthocyanin concentration of both cultivars was lower when exposed to high rather than low temperatures after véraison (the onset of ripening). However, the responses to temperature differed between the two cultivars, and anthocyanin accumulation could occur in 'Ruby Roman' at a higher temperature than in 'Aki Queen'. High temperatures increased the expression of VlMybA1-2 and VlMybA1-3, which encode myeloblastosis (MYB)-related transcription factors; however, the expression of the anthocyanin biosynthesis-related structural genes uridine diphosphate-d-glucose: flavonoid 3-O-glucosyltransferase, flavonoid 3'5' hydroxylase, and flavonoid O-methyltransferase at different temperatures did not correspond with that of the expression of MybAs. The concentration of ABA and its derivatives increased under high temperatures, but that of auxin and cytokinin decreased. The observation that high temperatures induced the accumulation of ABA and expression of VlMybA1s but not the expression of anthocyanin biosynthesis-related structural genes implied the operation of a mechanism different from up-regulation of anthocyanin synthesis by VlMybA1s in the temperature response of grape berries.


Assuntos
Ácido Abscísico/biossíntese , Antocianinas/biossíntese , Frutas/metabolismo , Reguladores de Crescimento de Planta/biossíntese , Vitis/metabolismo , Temperatura Baixa , Regulação da Expressão Gênica de Plantas/genética , Temperatura Alta , Redes e Vias Metabólicas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Transcriptoma , Vitis/genética , Vitis/fisiologia
9.
BMC Plant Biol ; 19(1): 169, 2019 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-31035916

RESUMO

BACKGROUND: Lycium ruthenicum Murray is an important economic plant in China and contains higher levels of anthocyanins in its fruits than other Lyciums. However, the genetic mechanism of anthocyanin production in this plant is unknown. RESULTS: Based on previous transcriptome analysis, LrAN2 and LbAN2, encoding MYB transcription factors, were isolated from L. ruthenicum and L. barbarum, respectively. Both genes contained two introns, encoded 257 amino acids with two-Aa difference, and carried the unabridged HTH-MYB, MYB-like DNA-binding, and SANT domains. In the phylogenetic trees, LrAN2 and LbAN2 were found to be closely related to NtAN2, which regulates anthocyanin biosynthesis in tobacco. Overexpression of LrAN2 and LbAN2 induced anthocyanin biosynthesis in all tissues of tobacco. The anthocyanin content in the leaves of transgenic lines with LbAN2 was lower than LrAN2. It indicated that the function of LbAN2 was weaker than LrAN2. The AN2 transcript could be detected only in the fruits of L. ruthenicum and increased during fruit development, accompanied by anthocyanin accumulation. In natural population, the alleles LrAN2 and LrAN2 were associated strictly with L. ruthenicum and L. barbarum, respectively. Moreover, an AN2 genetic diversity study suggested that Lyciums with yellow, white, purple, and jujube red fruits were derived from L. ruthenicum. CONCLUSIONS: Two AN2 alleles, from L. ruthenicum and L. barbarum, were functional MYB transcriptor regulating anthocyanin biosynthesis. The functional diversity and high expression level of LrAN2 could be the reason for high anthocyanin content in the fruit of L. ruthenicum. Lyciums with yellow, white, purple, and jujube red fruits were derived from L. ruthenicum based on AN2 sequence diversity. The results may be advantageous in identifying new varieties and breeding new cultivars.


Assuntos
Antocianinas/biossíntese , Genes de Plantas , Lycium/genética , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Alelos , China , Frutas/genética , Frutas/crescimento & desenvolvimento , Genes myb , Variação Genética , Lycium/crescimento & desenvolvimento , Lycium/metabolismo , Proteínas de Plantas/metabolismo , Fatores de Transcrição/metabolismo
10.
BMC Genomics ; 20(1): 353, 2019 May 09.
Artigo em Inglês | MEDLINE | ID: mdl-31072309

RESUMO

BACKGROUND: Bagging is commonly used to enhance red pigmentation and thereby improve fruit quality of apples (Malus domestica). The green-skinned apple cultivar 'Granny Smith' develops red pigmentation after bagging removal, but the signal transduction pathways mediating light-induced anthocyanin accumulation in apple peel are yet to be defined. The aim of this study was to identify the mechanisms underpinning red pigmentation in 'Granny Smith' after bag removal based on transcriptome sequencing. RESULTS: The anthocyanin content in apple peel increased considerably after bag removal, while only trace amounts of anthocyanins were present in the peel of unbagged and bagged fruits. RNA sequencing identified 18,152 differentially expressed genes (DEGs) among unbagged, bagged, and bag-removed fruits at 0, 4, and 10 days after bag removal. The DEGs were implicated in light signal perception and transduction, plant hormone signal transduction, and antioxidant systems. Weighted gene co-expression network analysis of DEGs generated a module of 23 genes highly correlated with anthocyanin content. The deletion of - 2026 to - 1870 bp and - 1062 to - 964 bp regions of the MdMYB1 (LOC103444202) promoter induced a significant decrease in glucuronidase activity and anthocyanin accumulation in apple peel. CONCLUSIONS: Bagging treatment can induce red pigmentation in 'Granny Smith' via altering the expression patterns of genes involved in crucial signal transduction and biochemical metabolic pathways. The - 2026 to - 1870 bp and - 1062 to - 964 bp regions of the MdMYB1 promoter are essential for MdMYB1-mediated regulation of anthocyanin accumulation in the 'Granny Smith' apple cultivar. The findings presented here provide insight into the mechanisms of coloration in the peel of 'Granny Smith' and other non-red apple cultivars.


Assuntos
Antocianinas/biossíntese , Frutas/genética , Perfilação da Expressão Gênica/métodos , Malus/genética , Frutas/metabolismo , Regulação da Expressão Gênica de Plantas , Malus/metabolismo , Pigmentação , Proteínas de Plantas/genética
11.
Food Chem ; 289: 112-120, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-30955592

RESUMO

Sucrose acts as a vital signal that modulates fruit ripening. In current study, 50 mM sucrose was applied in strawberry fruit to investigate the regulation of sucrose in anthocyanin synthesis after harvest. The results showed that sucrose treatment increased the contents of glucose, fructose and sucrose, which were 19.76%, 15.83% and 16.50% higher, respectively, compared with control at the end of storage. The increase of glucose and fructose contents resulted from the activation of acid invertase by sucrose treatment. In addition, sucrose treatment specifically increased four pelargonidin derivatives, pelargonidin 3-glucoside, pelargonidin 3-rutinoside, pelargonidin 3-malonylglucoside and pelargonidin 3-methylmalonyglucoside, during the storage. Further, transcriptional profiles and enzyme activities analysis revealed that the accumulation of pelargonidin derivatives was related to the activation of the pentose phosphate pathway, shikimate pathway, phenylpropanoid pathway, and flavonoid pathway. These results provided new insights into the regulation of sucrose on the accumulation of individual anthocyanins.


Assuntos
Antocianinas/biossíntese , Fragaria/metabolismo , Antocianinas/análise , Antocianinas/metabolismo , Flavonoides/metabolismo , Fragaria/química , Frutas/química , Frutas/metabolismo , Glucose/análise , Glucosefosfato Desidrogenase/genética , Glucosefosfato Desidrogenase/metabolismo , Espectrometria de Massas , Via de Pentose Fosfato/efeitos dos fármacos , Fenilalanina Amônia-Liase/metabolismo , Sacarose/farmacologia , Transcinamato 4-Mono-Oxigenase/metabolismo
12.
Biol Res ; 52(1): 17, 2019 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-30935421

RESUMO

BACKGROUND: Prunella vulgaris L. has been an important medicinal plant for the treatment of thyroid gland malfunction and mastitis in China for over 2000 years. There is an urgent need to select effective wavelengths for greenhouse cultivation of P. vulgaris as light is a very important factor in P. vulgaris growth. Here, we described the effects of natural light (control) and UV solar exclusion on the morphological and physiological traits, secondary metabolites contents and antioxidant activities of P. vulgaris. RESULTS: The results showed that UV solar exclusion resulted in remarkable alterations to morphological and biomass traits; significantly reduced the chlorophyll a, chlorophyll b and total chlorophyll contents; significantly enhanced the ratio of chlorophyll a to b; and significantly increased the carotenoid and anthocyanin contents in P. vulgaris. UV solar exclusion significantly increased the catalase (CAT) and peroxidase (POD) activities, increased superoxide dismutase (SOD) and ascorbate peroxidase (APX) activities and slightly decreased the glutathione (GSH) content. UV solar exclusion significantly increased the soluble sugar and H2O2 contents and increased the soluble protein content but significantly decreased the proline content and slightly decreased the MDA content. The secondary metabolite contents (total phenolics, rosmarinic acid, caffeic acid, hyperoside, ursolic acid and oleanolic acid) and in vitro antioxidative properties (DPPH· and ABTS·+scavenging activities) were significantly increased in P. vulgaris spicas under UV solar exclusion. Additionally, the total polysaccharide and total flavonoids contents were slightly increased by UV solar exclusion. The salviaflaside content was significantly reduced by UV solar exclusion. CONCLUSION: Our study demonstrated that P. vulgaris activates several antioxidant defence systems against oxidative damage caused by UV solar exclusion.


Assuntos
Antocianinas/biossíntese , Antioxidantes/metabolismo , Fotossíntese/fisiologia , Prunella/metabolismo , Antioxidantes/efeitos da radiação , Prunella/química , Prunella/efeitos da radiação , Raios Ultravioleta
13.
Plant Physiol Biochem ; 138: 130-139, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30870763

RESUMO

The flower color of Paeonia 'Coral Sunset' and 'Pink Hawaiian Coral' changes from coral to pink to pale yellow during flowering, which confers high ornamental value to these two cultivars. However, the molecular mechanism underlying flower color change is still unclear. In this study, flavonoids in petals of Paeonia 'Coral Sunset' and 'Pink Hawaiian Coral' at seven flowering stages were analyzed to explore the effects of the flavonoid component on changes in flower color. In addition, four cDNA libraries of 'Coral Sunset' during the critical blooming stages were constructed and the transcriptome was sequenced to investigate the molecular mechanism underlying changes to flower color. Two anthocyanins (cyanidin-3,5-di-O-glucoside and peonidin-3,5-di-O-glucoside) were detected in both cultivars. Total anthocyanin content in both cultivars accumulated continuously from stages 1-3 and then decreased sharply. Correlation analysis showed that the change in flower color from coral to pink to pale yellow is due to a significant decrease in anthocyanin content. A total of 91,583 Unigenes were obtained in 'Coral Sunset', 33,962 (37.08%) of which were annotated to major databases. Based on the enrichment analysis of differentially expressed genes, eight structural genes (CHS, F3H, F3'H, FLS, DRF, ANS, ANR and UFGT) and 13 transcription factors (five MYB, three bHLH, one WD40, one HY5, one PIF3, one COP1 and two PHY) related to flavonoid biosynthesis were screened. The qRT-PCR results were generally consistent with the high-throughput sequencing results. This research will provide a foundation to clarify the mechanisms underlying changes in flower color of herbaceous peony.


Assuntos
Antocianinas , Flores , Perfilação da Expressão Gênica , Genes de Plantas , Paeonia , Pigmentação/genética , Pigmentos Biológicos , Antocianinas/biossíntese , Antocianinas/genética , Flores/genética , Flores/metabolismo , Paeonia/genética , Paeonia/metabolismo , Pigmentos Biológicos/biossíntese , Pigmentos Biológicos/genética
14.
Int J Mol Sci ; 20(5)2019 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-30823465

RESUMO

Flower color is a charming phenotype with very important ornamental and commercial values. Anthocyanins play a critical role in determining flower color pattern formation, and their biosynthesis is typically regulated by R2R3-MYB transcription factors (TFs). Paeonia suffruticosa is a famous ornamental plant with colorful flowers. However, little is known about the R2R3-MYB TFs that regulate anthocyanin accumulation in P. suffruticosa. In the present study, two R2R3-MYB TFs, namely, PsMYB114L and PsMYB12L, were isolated from the petals of P. suffruticosa 'Shima Nishiki' and functionally characterized. Sequence analysis suggested that PsMYB114L contained a bHLH-interaction motif, whereas PsMYB12L contained two flavonol-specific motifs (SG7 and SG7-2). Subsequently, the in vivo function of PsMYB114L and PsMYB12L was investigated by their heterologous expression in Arabidopsis thaliana and apple calli. In transgenic Arabidopsis plants, overexpression of PsMYB114L and of PsMYB12L caused a significantly higher accumulation of anthocyanins, resulting in purple-red leaves. Transgenic apple calli overexpressing PsMYB114L and PsMYB12L also significantly enhanced the anthocyanins content and resulted in a change in the callus color to red. Meanwhile, gene expression analysis in A. thaliana and apple calli suggested that the expression levels of the flavonol synthase (MdFLS) and anthocyanidin reductase (MdANR) genes were significantly downregulated and the dihydroflavonol 4-reductase (AtDFR) and anthocyanin synthase (AtANS) genes were significantly upregulated in transgenic lines of PsMYB114L. Moreover, the expression level of the FLS gene (MdFLS) was significantly downregulated and the DFR (AtDFR/MdDFR) and ANS (AtANS/MdANS) genes were all significantly upregulated in transgenic lines plants of PsMYB12L. These results indicate that PsMYB114L and PsMYB12L both enhance anthocyanin accumulation by specifically regulating the expression of some anthocyanin biosynthesis-related genes in different plant species. Together, these results provide a valuable resource with which to further study the regulatory mechanism of anthocyanin biosynthesis in P. suffruticosa and for the breeding of tree peony cultivars with novel and charming flower colors.


Assuntos
Antocianinas/biossíntese , Paeonia/genética , Proteínas de Plantas/genética , Proteínas Proto-Oncogênicas c-myb/genética , Arabidopsis/genética , Arabidopsis/metabolismo , Regulação da Expressão Gênica de Plantas , Oxirredutases/genética , Oxirredutases/metabolismo , Paeonia/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Proteínas Proto-Oncogênicas c-myb/química , Proteínas Proto-Oncogênicas c-myb/metabolismo
15.
BMC Evol Biol ; 19(Suppl 1): 45, 2019 02 26.
Artigo em Inglês | MEDLINE | ID: mdl-30813913

RESUMO

BACKGROUND: The members of the Triticeae tribe are characterised by the presence of orthologous and homoeologous gene copies regulating flavonoid biosynthesis. Among transcription factors constituting a regulatory MBW complex, the greatest contribution to the regulation of flavonoid biosynthetic pathway is invested by R2R3-Myb-type TFs. Differently expressed R2R3-Myb copies activate the synthesis of various classes of flavonoid compounds in different plant tissues. The aim of this research was the identification, comparison and analysis of full-length sequences of the duplicated R2R3-Myb Mpc1 (Myb protein c1) gene copies in barley and wheat genomes. RESULTS: The Mpc1 genes were identified in homoeologous group 4 and 7 chromosomes: a total of 3 copies in barley (Hordeum vulgare L.) and 8 copies in bread wheat (Triticum aestivum L.) genomes. All Mpc1 genes have a similar two-exon structure, and almost all of them are transcriptionally active. The calculation of the divergence time revealed that first duplication between 4 and 7 chromosomes of the common ancestor of the Triticeae tribe occurred about 35-46 million years ago (MYA); the last duplication arised about 16-19 MYA before the divergence Triticum and Hordeum genera The connection between gene expression and the appearance of anthocyanin pigmentation was found for three genes from homoeologous group 4 chromosomes: TaMpc1-A2 (5AL) in wheat coleoptile, HvMpc1-H2 (4HL) in barley lemma and aleurone layer, and HvMpc1-H3 (4HL) in barley aleurone layer. TaMpc1-D4 (4DL) from the wheat genome showed a strong level of expression regardless of the colour of coleoptile or pericarp. It is assumed, that this gene regulates the biosynthesis of uncoloured flavonoids in analysed tissues. CONCLUSIONS: The regulatory R2R3-Myb genes involved in anthocyanin synthesis were identified and characterised in Triticeae tribe species. Genes designated HvMpc1-H2 and HvMpc1-H3 appeared to be the main factors underlying intraspecific variation of H. vulgare by lemma and aleurone colour. TaMpc1-A2 is the co-regulator of the Mpc1-1 genes in bread wheat genome controlling anthocyanin synthesis in coleoptile.


Assuntos
Hordeum/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Triticum/genética , Antocianinas/biossíntese , Cromossomos de Plantas/genética , Regulação da Expressão Gênica de Plantas , Genômica , Hordeum/metabolismo , Especificidade da Espécie , Triticum/metabolismo
16.
J Plant Physiol ; 236: 34-38, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30856405

RESUMO

The antioxidant properties of black rice are attributed to the high anthocyanin content in the pericarp. Light-dependent regulation of anthocyanin biosynthesis and the associated regulatory genes have been extensively studied in many plant species, including rice. Light is considered indispensable for anthocyanin accumulation in plants. Here, we report that anthocyanin biosynthesis and accumulation in the dark is negatively regulated by ethylene, as the ethylene biosynthesis inhibitor aminoethoxyvinylglycine hydrochloride (AVG)-treated samples in the dark exhibited significantly higher transcript levels of biosynthesis genes, including CHI, DFR, ANS, ANR, F3H, F'3H, CHS and UGFT, compared to untreated controls. Upregulation of these biosynthesis genes was accompanied by simultaneous de-repression of the R2-R3 domain and bHLH-containing transcription factors Kala3 and Kala4 at the transcript level. Additionally, bHLH152, which shows high similarity to Arabidopsis PIF3, is involved in the regulation of anthocyanin. These findings highlight the role of ethylene in modulating tissue-specific regulation of anthocyanin biosynthesis genes in the dark.


Assuntos
Antocianinas/metabolismo , Grão Comestível/metabolismo , Etilenos/metabolismo , Oryza/metabolismo , Reguladores de Crescimento de Planta/metabolismo , Antocianinas/biossíntese , Escuridão , Regulação da Expressão Gênica de Plantas , Reação em Cadeia da Polimerase em Tempo Real
17.
Molecules ; 24(5)2019 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-30866466

RESUMO

Red coleoptiles can help crops to cope with adversity and the key genes that are responsible for this trait have previously been isolated from Triticum aestivum, Triticum urartu, and Aegilops tauschii. This report describes the use of transcriptome analysis to determine the candidate gene that controls the trait for white coleoptiles in T. monococcum by screening three cultivars with white coleoptiles and two with red coleoptiles. Fifteen structural genes and two transcription factors that are involved in anthocyanin biosynthesis were identified from the assembled UniGene database through BLAST analysis and their transcript levels were then compared in white and red coleoptiles. The majority of the structural genes reflected lower transcript levels in the white than in the red coleoptiles, which implied that transcription factors related to anthocyanin biosynthesis could be candidate genes. The transcript levels of MYC transcription factor TmMYC-A1 were not significantly different between the white and red coleoptiles and all of the TmMYC-A1s contained complete functional domains. The deduced amino acid sequence of the MYB transcription factor TmMYB-A1 in red coleoptiles was homologous to TuMYB-A1, TaMYB-A1, TaMYB-B1, and TaMYB-D1, which control coleoptile color in corresponding species and contained the complete R2R3 MYB domain and the transactivation domain. TmMYB-a1 lost its two functional domains in white coleoptiles due to a single nucleotide deletion that caused premature termination at 13 bp after the initiation codon. Therefore, TmMYB-A1 is likely to be the candidate gene for the control of the red coleoptile trait, and its loss-of-function mutation leads to the white phenotype in T. monococcum.


Assuntos
Cotilédone/genética , Perfilação da Expressão Gênica/métodos , Fatores de Transcrição/genética , Triticum/anatomia & histologia , Antocianinas/biossíntese , Cotilédone/anatomia & histologia , Regulação da Expressão Gênica de Plantas , Sequenciamento de Nucleotídeos em Larga Escala , Mutação , Proteínas de Plantas/genética , Locos de Características Quantitativas , Análise de Sequência de RNA , Triticum/genética
18.
Plant Physiol Biochem ; 139: 273-282, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30925437

RESUMO

Anthocyanins in apple species are important secondary metabolites that are beneficial for human health. Previous studies revealed that methyl jasmonate (MeJA) promotes anthocyanin accumulation by up-regulating the transcription of related genes. In this study, we isolated a jasmonate (JA)-induced apple MYB gene, MdMYB24-like (MdMYB24L). The encoded nuclear protein contains a conserved R2R3 domain and is homologous to Arabidopsis thaliana AtMYB24. Additionally, MdMYB24L was observed to interact with JA signaling factors (MdJAZ8, MdJAZ11, and MdMYC2) in yeast and in planta. The MdMYC2 protein was also targeted by MdJAZ8 and MdJAZ11, which are rapidly degraded under MeJA treatment. The overexpression of MdMYB24L resulted in higher anthocyanin contents in the transgenic apple 'Orin' calli than in the wild-type control calli. Moreover, the expression levels of the anthocyanin biosynthesis structural genes MdUFGT and MdDFR were up-regulated in the transgenic calli. Furthermore, MdMYB24L positively regulated the transcription of MdDFR and MdUFGT by binding to the MYB-binding site motifs in their promoters. Interestingly, the interaction between MdMYC2 and MdMYB24L further enhanced the transcription of MdUFGT, whereas MdJAZ8 and MdJAZ11 attenuated this effect. We herein provide new details regarding the molecular mechanism by which MYB transcription factors help regulate anthocyanin biosynthesis via JA signaling pathways.


Assuntos
Acetatos/farmacologia , Antocianinas/biossíntese , Arabidopsis/metabolismo , Ciclopentanos/farmacologia , Malus/metabolismo , Oxilipinas/farmacologia , Fatores de Transcrição/metabolismo , Arabidopsis/efeitos dos fármacos , Proteínas de Arabidopsis/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Malus/efeitos dos fármacos
19.
Genome ; 62(4): 253-266, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30807237

RESUMO

Broccoli (Brassica oleracea var. italica L.) is a highly nutritious vegetable that typically forms pure green or purple florets. However, green broccoli florets sometimes accumulate slight purplish pigmentation in response environmental factors, decreasing their market value. In the present study, we aimed to develop molecular markers to distinguish broccoli genotypes as pure green or purplish floret color at the early seedling stage. Anthocyanins are known to be involved in the purple pigmentation in plants. The purplish broccoli lines were shown to accumulate purple pigmentation in the hypocotyls of very young seedlings; therefore, the expression profiles of the structural and regulatory genes of anthocyanin biosynthesis were analyzed in the hypocotyls using qRT-PCR. BoPAL, BoDFR, BoMYB114, BoTT8, BoMYC1.1, BoMYC1.2, and BoTTG1 were identified as putative candidate genes responsible for the purple hypocotyl color. BoTT8 was much more highly expressed in the purple than green hypocotyls; therefore, it was cloned and sequenced from various broccoli lines, revealing SNP and InDel variations between these genotypes. We tested four SNPs (G > A; A > T; G > C; T > G) in the first three exons and a 14-bp InDel (ATATTTATATATAT) in the BoTT8 promoter in 51 broccoli genotypes, and we found these genetic variations could distinguish the green lines, purple lines, and F1 hybrids. These novel molecular markers could be useful in broccoli breeding programs to develop a true green or purple broccoli cultivar.


Assuntos
Antocianinas/biossíntese , Brassica/genética , Hipocótilo/anatomia & histologia , Brassica/anatomia & histologia , Clonagem Molecular , DNA de Plantas , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Marcadores Genéticos , Hipocótilo/metabolismo , Pigmentação/genética , Polimorfismo de Nucleotídeo Único , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de DNA
20.
Food Chem ; 283: 404-413, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-30722891

RESUMO

The grapevine (Vitis vinifera L.) berry coloring mechanism in response to seasonal rain during grape ripening remains poorly understood. Therefore, anthocyanin biosynthesis regulation, dynamic changes in anthocyanin accumulation, biosynthetic enzyme activities, and related gene expression patterns were investigated in Cabernet Sauvignon grown under rain-shelter cultivation and open-field cultivation. Results showed that anthocyanin biosynthesis was strongly repressed during the rainy season. Environmental fluctuation from seasonal rain provoked metabolic responses in grapes, and there was a significantly greater accumulation of most of the anthocyanins, mainly the compositions of non-acylated and non-methylated, under rain-shelter cultivation; these findings indicate that rain-shelter cultivation may help improve tolerance to seasonal rain-induced stresses. Obvious resilience was observed in anthocyanins of open-field-cultivated grapes at harvest. Hierarchical cluster analysis indicated strong correlations between anthocyanin contents, CHI and DFR activities, and VvMYB5b transcriptional level. These findings provide novel insight into the crucial factors that directly modulate anthocyanin biosynthesis and consequently control grape coloration.


Assuntos
Agricultura/métodos , Antocianinas/biossíntese , Vitis/química , Antocianinas/análise , Cromatografia Líquida de Alta Pressão , Análise por Conglomerados , Análise Discriminante , Proteínas de Plantas/genética , Chuva , Estações do Ano , Transcrição Genética , Vitis/crescimento & desenvolvimento , Vitis/metabolismo
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