Pelargonidin Derivatives Research Articles

Allele dosage–dependent selection of recessive F3’H allele homozygote altered anthocyanin composition in sweetpotato

To change the major anthocyanin in the storage roots of sweetpotato (Ipomoea batatas) from purplish cyanidin derivatives to reddish pelargonidin derivatives, the dosage of a recessive allele in the F3’H gene locus was enriched by successive crossing and allele dosage estimation. A putative recessive allele of the F3’H gene, named f3’htm, was found in the cultivar Tanegeshimamurasaki and contained a 57-bp deletion causing a premature stop codon. Starting with the parents having a simplex f3’htm allele, plants considered to be homozygous with f3’htm were obtained in the F4 generation. The storage roots and other pigmented tissues of the homozygous plants showed a reddish color, whereas the original parents or F4 plants without homozygous f3’htm were purplish. More than approximately 90% of the anthocyanins in the storage roots of plants with homozygous f3’htm were pelargonidin derivatives. These results suggest that allele dosage–dependent selection is also possible in autohexaploid crops to develop mutants with novel phenotypes. In addition, the obtained plants with homozygous f3’htm can be important materials for breeding sweetpotato cultivars for pelargonidin production.

Hybrid-Transcriptome Sequencing and Associated Metabolite Analysis Reveal Putative Genes Involved in Flower Color Difference in Rose Mutants.

Gene mutation is a common phenomenon in nature that often leads to phenotype differences, such as the variations in flower color that frequently occur in roses. With the aim of revealing the genomic information and inner mechanisms, the differences in the levels of both transcription and secondary metabolism between a pair of natural rose mutants were investigated by using hybrid RNA-sequencing and metabolite analysis. Metabolite analysis showed that glycosylated derivatives of pelargonidin, e.g., pelargonidin 3,5 diglucoside and pelargonidin 3-glucoside, which were not detected in white flowers (Rosa ‘Whilte Mrago Koster’), constituted the major pigments in pink flowers. Conversely, the flavonol contents of petal, such as kaempferol-3-glucoside, quercetin 3-glucoside, and rutin, were higher in white flowers. Hybrid RNA-sequencing obtained a total of 107,280 full-length transcripts in rose petal which were annotated in major databases. Differentially expressed gene (DEG) analysis showed that the expression of genes involved in the flavonoid biosynthesis pathway was significantly different, e.g., CHS, FLS, DFR, LDOX, which was verified by qRT-PCR during flowering. Additionally, two MYB transcription factors were found and named RmMYBAN2 and RmMYBPA1, and their expression patterns during flowering were also analyzed. These findings indicate that these genes may be involved in the flower color difference in the rose mutants, and competition between anthocyanin and flavonol biosynthesis is a primary cause of flower color variation, with its regulation reflected by transcriptional and secondary metabolite levels.

Metabolite and gene expression analysis reveal the molecular mechanism for petal colour variation in six Centaurea cyanus cultivars

Centaurea cyanus is a popular garden plant native to Europe. Although their petals show abundant colour variations, the flavonoid profiling and the potential molecular mechanisms remain unclear. In the present study, we collected six cornflower cultivars with white, pink, red, blue, mauve and black petals. Ultra-performance liquid chromatography coupled with photodiode array and tandem mass spectrometry (UPLC-MS/MS) was used to investigate the comparative profiling of flavonoids both qualitatively and quantitatively. Ten anthocyanins, six flavones and two flavonols were separated and putatively identified. Except for white petals without any anthocyanins, both pink and red flowers contained pelargonidin derivatives, whereas blue, mauve and black petals accumulated cyanidins. The expression patterns of genes involved in the flavonoid biosynthesis were performed by real-time quantitative reverse transcription-PCR. The anthocyanin biosynthetic pathway in white petals was inhibited starting from flavanone 3-hydroxylase, resulting in the absence of anthocyanin accumulation. The open reading frame of flavonoid 3′-hydroxylase in pink and red petals was truncated; this led to loss of a haem binding site, a conserved motif in the cytochrome P450 family, and loss of conversion from dihydrokaempferol to dihydroquercetin. The significantly higher expression of structural genes corresponding to the hyper-accumulation of flavonoids in black petals may play an important role in black coloration. Remarkably, the mauve and blue petals accumulated the same cyanidin derivative but contained apigenin with different modifications on the 4′ position, which may cause the coloration differences. The results obtained in this study will provide insights into the mechanisms of vivid colour diversities in cornflower.

Effect of exogenous sucrose on anthocyanin synthesis in postharvest strawberry fruit

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.

Differential accumulation of pelargonidin glycosides in petals at three different developmental stages of the orange-flowered gentian (Gentiana lutea L. var. aurantiaca).

Corolla color in Gentiana lutea L. exhibits a yellow/orange variation. We previously demonstrated that the orange petal color of G. lutea L. var. aurantiaca is predominantly caused by newly synthesized pelargonidin glycosides that confer a reddish hue to the yellow background color, derived from the carotenoids. However, the anthocyanin molecules of these pelargonidin glycosides are not yet fully identified and characterized. Here, we investigated the regulation, content and type of anthocyanins determining the petal coloration of the orange-flowered G. lutea L. var. aurantiaca. Anthocyanins from the petals of G. lutea L. var. aurantiaca were characterized and quantified by HPLC-ESI-MS/MS (High-performance liquid chromatography-electrospray ionization-tandem mass spectrometry) coupled with a diode array detector in flowers at three different stages of development (S1, S3 and S5). Eleven pelargonidin derivatives were identified in the petals of G. lutea L. var. aurantiaca for the first time, but quantitative and qualitative differences were observed at each developmental stage. The highest levels of these pelargonidin derivatives were reached at the fully open flower stage (S5) where all anthocyanins were detected. In contrast, not all the anthocyanins were detected at the budlet stage (S1) and mature bud stage (S3) and those corresponded to more complex pelargonidin derivatives. The major pelargonidin derivatives found at all the stages were pelargonidin 3-O-glucoside, pelargonidin 3,5-O-diglucoside and pelargonidin 3-O-rutinoside. Furthermore, the expression of DFR (dihydroflavonol 4-reductase), ANS (anthocyanidin synthase), 3GT (UDP-glucose:flavonoid 3-O-glucosyltransferase), 5GT (UDP-glucose:flavonoid 5-O-glucosyltransferase) and 5AT (anthocyanin 5-aromatic acyltransferase) genes was analyzed in the petals of three developmental stages, showing that the expression level of DFR, ANS and 3GT parallels the accumulation of the pelargonidin glucosides. Overall, this study enhances the knowledge of the biochemical basis of flower coloration in Gentiana species, and lays a foundation for breeding of flower color and genetic variation studies on Gentiana varieties.

The rare orange-red colored Euphorbia pulcherrima cultivar \u2018Harvest Orange\u2019 shows a nonsense mutation in a flavonoid 3\u2019-hydroxylase allele expressed in the bracts

BackgroundCommercially available poinsettia (Euphorbia pulcherrima) varieties prevalently accumulate cyanidin derivatives and show intense red coloration. Orange-red bract color is less common. We investigated four cultivars displaying four different red hues with respect to selected enzymes and genes of the anthocyanin pathway, putatively determining the color hue.ResultsRed hues correlated with anthocyanin composition and concentration and showed common dark red coloration in cultivars ‘Christmas Beauty’ and ‘Christmas Feeling’ where cyanidin derivatives were prevalent. In contrast, orange-red bract color is based on the prevalent presence of pelargonidin derivatives that comprised 85% of the total anthocyanin content in cv. ‘Premium Red’ and 96% in cv. ‘Harvest Orange’ (synonym: ‘Orange Spice’). cDNA clones of flavonoid 3′-hydroxylase (F3′H) and dihydroflavonol 4-reductase (DFR) were isolated from the four varieties, and functional activity and substrate specificity of the corresponding recombinant enzymes were studied. Kinetic studies demonstrated that poinsettia DFRs prefer dihydromyricetin and dihydroquercetin over dihydrokaempferol, and thus, favor the formation of cyanidin over pelargonidin. Whereas the F3′H cDNA clones of cultivars ‘Christmas Beauty’, ‘Christmas Feeling’, and ‘Premium Red’ encoded functionally active enzymes, the F3′H cDNA clone of cv. ‘Harvest Orange’ contained an insertion of 28 bases, which is partly a duplication of 20 bases found close to the insertion site. This causes a frameshift mutation with a premature stop codon after nucleotide 132 and, therefore, a non-functional enzyme. Heterozygosity of the F3′H was demonstrated in this cultivar, but only the mutated allele was expressed in the bracts. No correlation between F3′H-expression and the color hue could be observed in the four species.ConclusionsRare orange-red poinsettia hues caused by pelargonidin based anthocyanins can be achieved by different mechanisms. F3′H is a critical step in the establishment of orange red poinsettia color. Although poinsettia DFR shows a low substrate specificity for dihydrokaempferol, sufficient precursor for pelargonidin formation is available in planta, in the absence of F3’H activity.

Great Cause-Small Effect: Undeclared Genetically Engineered Orange Petunias Harbor an Inefficient Dihydroflavonol 4-Reductase.

A recall campaign for commercial, orange flowering petunia varieties in spring 2017 caused economic losses worldwide. The orange varieties were identified as undeclared genetically engineered (GE)-plants, harboring a maize dihydroflavonol 4-reductase (DFR, A1), which was used in former scientific transgenic breeding attempts to enable formation of orange pelargonidin derivatives from the precursor dihydrokaempferol (DHK) in petunia. How and when the A1 cDNA entered the commercial breeding process is unclear. We provide an in-depth analysis of three orange petunia varieties, released by breeders from three countries, with respect to their transgenic construct, transcriptomes, anthocyanin composition, and flavonoid metabolism at the level of selected enzymes and genes. The two possible sources of the A1 cDNA in the undeclared GE-petunia can be discriminated by PCR. A special version of the A1 gene, the A1 type 2 allele, is present, which includes, at the 3′-end, an additional 144 bp segment from the non-viral transposable Cin4-1 sequence, which does not add any functional advantage with respect to DFR activity. This unequivocally points at the first scientific GE-petunia from the 1980s as the A1 source, which is further underpinned e.g., by the presence of specific restriction sites, parts of the untranslated sequences, and the same arrangement of the building blocks of the transformation plasmid used. Surprisingly, however, the GE-petunia cannot be distinguished from native red and blue varieties by their ability to convert DHK in common in vitro enzyme assays, as DHK is an inadequate substrate for both the petunia and maize DFR. Recombinant maize DFR underpins the low DHK acceptance, and, thus, the strikingly limited suitability of the A1 protein for a transgenic approach for breeding pelargonidin-based flower color. The effect of single amino acid mutations on the substrate specificity of DFRs is demonstrated. Expression of the A1 gene is generally lower than the petunia DFR expression despite being under the control of the strong, constitutive p35S promoter. We show that a rare constellation in flavonoid metabolism—absence or strongly reduced activity of both flavonol synthase and B-ring hydroxylating enzymes—allows pelargonidin formation in the presence of DFRs with poor DHK acceptance.

Inhibition of α-glucosidase, α-amylase, and aldose reductase by potato polyphenolic compounds.

Diabetes mellitus is a chronic disease that is becoming a serious global health problem. Diabetes has been considered to be one of the major risks of cataract and retinopathy. Synthetic and natural product inhibitors of carbohydrate degrading enzymes are able to reduce type 2 diabetes and its complications. For a long time, potatoes have been portrayed as unhealthy for diabetic patients by some nutritionist due to their high starch content. However, purple and red potato cultivars have received considerable attention from consumers because they have high levels of polyphenolic compounds that have potent antioxidant activities. In this study, we screened the total phenolics (TP) and total anthocyanins (TA) and analyzed the phenolic and anthocyanin compounds in selected potato cultivars and advanced selections with distinct flesh colors (purple, red, yellow and white). Purple and red potato cultivars had higher levels of TP and TA than tubers with other flesh colors. Chlorogenic acid is the predominant phenolic acid, and major anthocyanin is composed of the derivatives of petunidin, peonidin, malvidin and pelargonidin. We tested the potential inhibitory effect of potato extracts on the activities of α-amylase and α-glucosidase, which were targeted to develop antidiabetic therapeutic agents. We also measured inhibitory effect of potato extracts on aldose reductase (AR) which is a key enzyme that has been a major drug target for the development of therapies to treat diabetic complications. Purple flesh tubers extract showed the most effective inhibition of α-amylase, α-glucosidase, and aldose reductase with IC50 values 25, 42, and 32 μg/ml, respectively. Kinetic studies showed that anthocyanins are noncompetitive inhibitors of these enzymes, whereas phenolic acids behaved as mixed inhibitors for α-amylase and α-glucosidase and noncompetitive inhibitors for AR. This study supports the development of a positive and healthful image of potatoes, which is an important issue for consumers.

Dihydroflavonol 4-Reductase Genes from Freesia hybrida Play Important and Partially Overlapping Roles in the Biosynthesis of Flavonoids.

Dihydroflavonol-4-reductase (DFR) is a key enzyme in the reduction of dihydroflavonols to leucoanthocyanidins in both anthocyanin biosynthesis and proanthocyanidin accumulation. In many plant species, it is encoded by a gene family, however, how the different copies evolve either to function in different tissues or at different times or to specialize in the use of different but related substrates needs to be further investigated, especially in monocot plants. In this study, a total of eight putative DFR-like genes were firstly cloned from Freesia hybrida. Phylogenetic analysis showed that they were classified into different branches, and FhDFR1, FhDFR2, and FhDFR3 were clustered into DFR subgroup, whereas others fell into the group with cinnamoyl-CoA reductase (CCR) proteins. Then, the functions of the three FhDFR genes were further characterized. Different spatio-temporal transcription patterns and levels were observed, indicating that the duplicated FhDFR genes might function divergently. After introducing them into Arabidopsis dfr (tt3-1) mutant plants, partial complementation of the loss of cyanidin derivative synthesis was observed, implying that FhDFRs could convert dihydroquercetin to leucocyanidin in planta. Biochemical assays also showed that FhDFR1, FhDFR2, and FhDFR3 could utilize dihydromyricetin to generate leucodelphinidin, while FhDFR2 could also catalyze the formation of leucocyanidin from dihydrocyanidin. On the contrary, neither transgenic nor biochemical analysis demonstrated that FhDFR proteins could reduce dihydrokaempferol to leucopelargonidin. These results were consistent with the freesia flower anthocyanin profiles, among which delphinidin derivatives were predominant, with minor quantities of cyanidin derivatives and undetectable pelargonidin derivatives. Thus, it can be deduced that substrate specificities of DFRs were the determinant for the categories of anthocyanins aglycons accumulated in F. hybrida. Furthermore, we also found that the divergence of the expression patterns for FhDFR genes might be controlled at transcriptional level, as the expression of FhDFR1/FhDFR2 and FhDFR3 was controlled by a potential MBW regulatory complex with different activation efficiencies. Therefore, it can be concluded that the DFR-like genes from F. hybrida have diverged during evolution to play partially overlapping roles in the flavonoid biosynthesis, and the results will contribute to the study of evolution of DFR gene families in angiosperms, especially for monocot plants.

Red-purple flower color and delphinidin-type pigments in the flowers of Pueraria lobata (Leguminosae)

A previously undescribed acylated anthocyanin was extracted from the red-purple flowers of Pueraria lobata with 5% HOAc-H2O, and determined to be petunidin 3-O-(β-glucopyranoside)-5-O-[6-O-(malonyl)-β-glucopyranoside], by chemical and spectroscopic methods. In addition, two known acylated anthocyanins, delphinidin 3-O-(β-glucopyranoside)-5-O-[6-O-(malonyl)-β-glucopyranoside] and malvidin 3-O-(β-glucopyranoside)-5-O-[6-O-(malonyl)-β-glucopyranoside] were identified. Delphinidin 3,5-di-glucoside, petunidin 3,5-di-glucoside, and malvidin 3,5-di-glucoside, have been known as major components of P. lobata in the former study. However, malonyl esters amounts were detected over 10 times compared with non-malonyl esters amounts. In those anthocyanins the most abundant anthocyanin was petunidin 3-O-(β-glucopyranoside)-5-O-[6-O-(malonyl)-β-glucopyranoside] in total flowers.On the visible absorption spectral curve of fresh red-purple petals, one characteristic absorption maximum was observed at 520 nm, which is similar to those of flowers containing pelargonidin derivatives. In contrast, the absorption spectral curve of old violet petals was observed at 500(sh), 536, 564(sh), and 613(sh) nm, which are similar to those of violet flowers containing delphinidin-type pigments. Pressed juices of both fresh red-purple petals and old violet petals had pH5.2 and 5.5 respectively, and had the same flavonoid constitution. Crude fresh red-purple petal pigments extracted by pH 2.2 and pH 5.2 buffers exhibited the same color and spectral curves as fresh red-purple petals and old violet petals, respectively. Moreover, in a cross-TLC experiment of crude extracted pigments, red-purple color was exhibited by the anthocyanin region and the crossed region of anthocyanins and isoflavone. Thus, it may be assumed that the unusually low pH in the vacuole of fresh petals plays an important role to form red-purple flower color against weak acidic pH in the vacuole of old violet P. lobata petals.

Identification of flavonoid 3′-hydroxylase in the yellow flower of Delphinium zalil

The flowers of delphinium cultivars owe their coloration to anthocyanins such as delphinidin or pelargonidin derivatives. To date, no delphinium cultivars have been found with red flowers due to the presence of cyanidin derivatives. This suggests that delphiniums do not have cyanidin biosynthesis ability because of the loss of function of flavonoid 3′ hydroxylase (F3′H). Here, we show that the wild delphinium species Delphinium zalil (synonym semibarbatum) can accumulate quercetin 3-glucosides in its sepals, presumably through F3′H activity. We isolated F3′H cDNA from D. zalil (DzF3′H) and produced a recombinant enzyme from a yeast transformant. The recombinant DzF3′H protein could convert naringenin, apigenin, dihydrokaempferol and kaempferol to eriodictyol, luteolin, dihydroquercetin and quercetin, respectively. An expression analysis confirmed that blue flowered D. grandiflorum does not express F3′H, and also showed that flavonoid 3′,5′-hydroxylase and anthocyanidin synthase do not function in D. zalil sepals. DzF3′H can act as a flavonoid hydroxylase to produce cyanidin accumulation. The introduction of the DzF3′H gene into other delphinium species by conventional breeding may enable development of cultivars with novel flower colors.

Co-pigmentation of pelargonidin derivatives in strawberry and red radish model solutions by the addition of phenolic fractions from mango peels

Pelargonidin-based colors suffer from notorious instability. A phenolic mango peel extract and defined phenolic fractions thereof were shown to effectively modulate the visible absorption of anthocyanins from strawberry (Fragaria x ananassa Duch.) and red radish (Raphanus sativus L.) by intermolecular co-pigmentation. Consistently, non-acylated pelargonidin derivatives from strawberry exerted significantly greater hyper- and bathochromic spectral shifts than their acylated counterparts from red radish. The addition of low molecular-weight co-pigments such as gallic acid and monogalloyl glucoses to strawberry anthocyanins led to strong hyperchromic shifts from 30% to 48%, while gallotannins (>six galloyl units) exerted smaller co-pigmentation effects (36±2%; Δλmax 13nm), possibly due to steric hindrances. In contrast, penta- and hexa-O-galloyl-glucose induced greatest and most stable co-pigmentation effects (53±2%; Δλmax 13nm). Irrespective of the underlying mechanisms and the responsible compounds, phenolic mango peel extracts might represent suitable color enhancers for coloring foodstuff, particularly for those containing non-acylated pelargonidin derivatives.

Genetics and Anthocyanin Analysis of Flower Color in Mexican Petunia

The genetics and anthocyanins responsible for flower color were studied in Ruellia simplex Wright (mexican petunia). An F2 population with 153 individuals segregating for four flower colors was developed from a cross between a maternal individual with white corolla with purple throat (WP) and a paternal individual with pink corolla (PK). All the F1 generation had fully purple flowers (P). The F2 generation segregated 94 P:30 PK:24 WP:5 WPK (WPK is a new color combination of white corolla limb and pink throat). These data were separated into groups for corolla limb color and for throat color. The ratio for corolla limb color segregated 94 P:30 PK:29 W, which fits a 9:3:4 recessive epistasis interaction (P = 0.22). The data for corolla throat segregated 118 P:35 PK, which fits a 3:1 ratio (P = 0.54). High-performance liquid chromatography mass spectrometry analyses were performed to elucidate the anthocyanins responsible for the four obtained flower colors. We found that delphinidin derivatives conferred purple corolla color, whereas pelargonidin derivatives were responsible for the pink corolla color. Purple corolla throat color was the result of delphinidin derivatives, whereas the pink color was the result of peonidin derivatives.

FURTHER RESEARCH CONCERNING THE EXTRACTION OF ANTHOCYANINS FROM STRAWBERRIES (FRAGARIA ×ANANASSA) AND ANALYSIS USING HPLC/Q-TOF MASS SPECTROMETRY

Anthocyanins were extracted from strawberries (Fragaria A—ananassa ‘Tochiotome’) using eight types of extraction media. The extracts were analyzed using high performance liquid chromatography-mass spectrometry. The quantity of anthocyanins recovered was influenced by the concentration and composition of the extracting solutions. Lower levels of anthocyanins were extracted using various concentrations of MeOH, ethanol, and aqueous HCl, while the use of 45% formic acid, 45% formic acid in MeOH, 45% formic acid in ethanol, or 1% HCl in MeOH or ethanol resulted in much higher extraction yields. Extraction using solutions containing formic acid yielded 16 anthocyanins , in contrast to other solutions from which only 7-10 anthocyanins were isolated. Pelargonidin (Pg) derivatives were the primary anthocyanins recovered. Ten Pg derivatives were detected, with the most abundant being Pg 3-glucoside, followed by Pg 3-rutinoside and Cyanidin 3-glucoside. We found that aqueous solutions containing more than 10% formic acid were the most productive in terms of extraction yield.

Antioxidant and pro-oxidant properties of acylated pelargonidin derivatives extracted from red radish (Raphanus sativus var. niger, Brassicaceae)

The antioxidant and pro-oxidant potential of an extract from red radish, in which the major compounds were acylated pelargonidin derivatives, were assessed with a variety of assays in vitro. The extract appeared to form a complex with Fe3+ or Cu2+. It displayed a concentration-dependant reducing power (1.16OD700 nm at a concentration of 4mM) and scavenging effect against 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radicals (with IC50=1.74±0.03mM). It could promote the cleavage of plasmid DNA with Cu(II)/H2O2 or Cu(II) alone. This DNA damage could be inhibited by horseradish peroxidase, catalase, and EDTA, respectively. The extract also showed growth inhibition of Bel-7402 cells at lower concentration. The results suggested that the formation of reactive oxygen species might be involved in the mechanism of DNA damage. The acylated pelargonidin derivatives extracted from red radish could act as antioxidant and pro-oxidant and their antioxidant and pro-oxidant properties were relative to the reaction conditions. It might provide novel antioxidant and anticarcinogenic agents.

Influence of the Genotype on the Anthocyanin Composition, Antioxidant Capacity and color of Chilean Pomegranate (Punica granatum L.) Juices

The bioactive compounds content in pomegranate (Punica granatum L.) juice is variable and depends on agronomic factors, genetic and fruit maturity. In Chile, the areas with this crop have increased more than 65% in the last 5 years. This paper aims to determine the phenolic composition, antioxidant capacity and chromatic parameters of juices from eight Chilean pomegranate genotypes (PG2 to PG9), containing red (PG2 to PG6) or pink arils (PG7 to PG9). Color, total polyphenol content (using Folin-Ciocalteu reagent), total anthocyanins (by differences in absorbance at different pHs of the juice) and antioxidant capacity (oxygen-radical absorbance capacity, ORAC) were determined. Anthocyanins were identified and quantify using HPLC-DAD and HPLC/ESI-MS, the results were expressed as cyanidin-3-glucoside. ANOVA was used for establish differences between total polyphenols (TP), antioxidant capacity, total anthocyanins (TA), total polyphenols/total anthocyanins relations (TP/TA) and colour parameters. Statgraphics plus 7.0 was used. Genotypes PG2-PG6 (red arils) showing higher values for total polyphenols (10551280 mg L -1 ) and anthocyanins (588-1328 mg L -1 ) and higher ORAC values (21.2-24.4 μmol Trolox mL -1 ) than the pink arils PG7-PG9 (12.7-16.8 μmol Trolox mL -1 ) values. The latter genotypes showed the highest values of pH (3.5-3.7). The anthocyanin compounds identified in the juices were the 3-glucoside and 3,5-diglucoside derivatives of delphinidin, cyanidin and pelargonidin. Differences in their relative amounts were found among the Chilean pomegranate genotypes studied. The relationships between TP/TA and Pg/Cy (pelargonidin 3-glucoside and pelargonidine 3,5-diglucoside/cyanidin 3-glucoside and cyanidin 3,5-diglucoside) could be useful in the analytical characterization of Chilean pomegranate genotypes, as well as in the differentiation with other juices containing anthocyanins.

Characterisation of major taste and health-related compounds of four strawberry genotypes grown at different Swiss production sites

Individual sugars, organic acids, anthocyanins and vitamin C were quantified in strawberry fruits of four newly-bred cultivars grown at two production sites in Switzerland with different soil, climatic conditions and altitudes (1060 and 480 m above sea level). All the measured compounds were significantly influenced by genotype. Pelargonidin-3-glucoside was the main anthocyanin present in all cultivars, while the presence of other pelargonidin derivatives was genotype-dependent. Differences of about 2-fold were observed among the studied cultivars for their vitamin C content. In the mountain region, where plants produced a higher fruit yield over a shorter period, the concentration of both health and taste-related compounds was detrimentally affected. In particular, the vitamin C content in the fruits was negatively related to the average yield per day. However, the compositional variations of strawberry fruits in response to different production sites were genotype specific. Within the four cultivars studied, cv. Antea was most affected by the production site, showing generally lower contents of all analysed compounds when cultivated at higher altitudes, whereas cv. Clery seemed to have the more consistent chemical composition, regardless of production site. The results presented in this work corroborate the dominant role of strawberry genotype over environmental factors.

2,4-Dichlorophenoxyacetic Acid Increases Peonidin Derivatives in Red Norland Periderm

Studies were conducted to determine the effects of selected plant growth regulators on red color development of Red Norland potato periderm, and the mechanism by which color changes occurred. Two derivatives of the auxin 2,4-dichlorophenoxyacetic acid (2,4-D ester and 2,4-D amine), S-abscisic acid, n-propyl dihydrojasmonate, and the ethylene synthesis inhibitor aminoethoxyvinylglycine were foliar applied at 4, 6, and 8 weeks after emergence. Only the 2,4-D ester and 2,4-D amine significantly improved tuber periderm red color in two seasons of evaluation. Red color faded during storage regardless of treatment. Anthocyanin content at vine kill was significantly increased by 2,4-D treatment in 2004 but not 2005. The 2,4-D treatments increased the ratio of peonidin derivatives to pelargonidin derivatives compared with controls. This change in ratio may explain the observed change in hue angle with 2,4-D treatment compared to controls.

Characterization of dihydroflavonol 4-reductases for recombinant plant pigment biosynthesis applications

Anthocyanins are colorful plant pigments with promising applications as pharmaceuticals and colorants. In order to engineer efficient pigment biosynthesis in Escherichia coli, the activities of various dihydroflavonol 4-reductases (DFRs) were characterized for the three primary dihydroflavonol substrates. The biochemical assays demonstrated variable DFR activities for dihydroflavonol with one B-ring hydroxyl group, the precursor of pelargonidin derivatives. In contrast, dihydroflavonols with two and three B-ring hydroxylation were metabolized with comparable efficiency. Furthermore, the catalysis of DFR for the secondary substrates, flavanones, also depended on the number of B-ring hydroxyl groups. Engineering the expression of the DFR clones together with plant-specific 4-coumaroyl:CoA ligase, chalcone synthase, chalcone isomerase, and flavanone 3-hydroxylase in E. coli resulted in the synthesis of pelargonidin at various levels, from p-coumaric acids. The identification of a robust DFR from this study can also be used for engineering recombinant synthesis of other bioactive flavonoids, such as flavan-3-ols.

Strawberry pelargonidin glycosides are excreted in urine as intact glycosides and glucuronidated pelargonidin derivatives in rats

Anthocyanins are natural dietary pigments with a wide array of biological properties that are possibly involved in the prevention of various diseases. These properties depend on their absorption and metabolism in the body. In the present study we first examined the gastric and intestinal absorption of pelargonidin 3-glucoside (Pg 3-glc) using rat in situ models. A high proportion of Pg 3-glc was rapidly absorbed from both the stomach (23 %) and small intestine (24 %). Its metabolism was further studied by feeding rats during 8 d with a diet enriched in freeze-dried strawberries. Only low amounts of total anthocyanins were recovered in 24 h urine (0.163 (SEM 0.013) % of ingested anthocyanins; n 8). Strawberry anthocyanins were analysed in urine by HPLC-electrospray ionisation-tandem MS. Similar proportions of intact glycosides (about 53 %) and glucuronidated metabolites (about 47 %) were found. Pg 3-glc was thus glucuronidated to a larger extent than cyanidin 3-glucoside. These results highlight the influence of the aglycone structure on anthocyanin metabolism.