Fruit Anthocyanin Research Articles

Transcriptional analysis in high-anthocyanin tomatoes reveals synergistic effect of Aft and atv genes

Anthocyanins are high value plant antioxidants, which are not present in the fruits of the cultivated tomato. However, both the dominant gene Anthocyanin fruit ( Aft) and the recessive gene atroviolacea ( atv), when introgressed into the domesticated tomato from two different wild Solanum species, stimulate a limited anthocyanin pigmentation. Surprisingly, the double mutant Aft/ Aft atv/ atv gives rise to intensely purple pigmented tomatoes. A transcript profiling analysis was carried out using quantitative RT-PCR and GeneChip ® Tomato Genome Arrays to identify differentially expressed genes when comparing Ailsa Craig, Aft/ Aft, atv/ atv, and Aft/ Aft atv/ atv fruits. Anthocyanin levels and the expression of the genes involved in anthocyanin production and compartmentalization were higher in the peel of Aft/ Aft atv/ atv fruits than in the individual parental lines. Moreover, a synergistic effect of the two alleles Aft and atv on the transcription of specific anthocyanin genes and the activation of the whole anthocyanin pathway was observed. Among the differentially expressed transcripts, genes involved in the phenylpropanoid pathway, biotic and abiotic stress responses, cell wall and hormone metabolism were over-represented in Aft/ Aft atv/ atv fruit peel. Transcriptomic analyses thus revealed that the activation of anthocyanin synthesis in the peel of tomato fruit was accompanied by a complex remodulation of gene expression.

Variation in anthocyanins and total phenolics of black raspberry populations

Black raspberry ( Rubus occidentalis L.) has long been recognized as a rich source of anthocyanins. Despite renewed interest in this crop for its potential health benefits, the range of variation in anthocyanin content and other phenolic compounds has not been well examined. Here we present anthocyanin concentration and profiles, as well as total phenolics, in the fruit of 26 black raspberry seedling populations (190 samples over two growing seasons) derived from cultivated and wild parents. There was more than a twofold difference in total anthocyanin concentration between the lowest and highest pigmented populations (ranging from 244.8 to 541.3 mg 100 mL −1). The relative amounts of the two major anthocyanins (cyanidin-3-rutinoside and cyanidin-3-xylosylrutinoside) in black raspberry fruit were significantly different. The range of total phenolics found was much lower (206.7–330.4 mg 100 mL −1). This information will provide a valuable baseline for researchers interested in studying the health effects of these compounds, product developers in the nutraceutical market, and breeders interested in developing new cultivars with improved fruit chemistry traits.

Effect of BTH on antioxidant enzymes, radical-scavenging activity and decay in strawberry fruit

The effect of benzo-thiadiazole-7-carbothioic acid S-methyl ester (BTH), at 0.2 g/l, on antioxidant enzymes, radical-scavenging activity and decay development in strawberry ( Fragaria × ananassa Duch.) fruit, was investigated. The results showed that BTH treatment enhanced the activities of antioxidant enzymes, including superoxide dismutase, ascorbate peroxidase, and glutathione reductase. The treatment increased the contents of phenolic and anthocyanin in strawberry fruit, as well as radical-scavenging capacity, expressed as 1,1-diphenyl-2-picrylhydrazyl and superoxide radical values and reducing power. The treatment also decreased the development of decay compared to control fruit. These results indicated that BTH might increase the disease resistance of strawberry fruits by enhancing their antioxidant systems and their free radical-scavenging capabilities.

Direct profiling of saccharides, organic acids and anthocyanins in fruits using electrospray droplet impact/secondary ion mass spectrometry

Electrospray droplet impact (EDI)/secondary ion mass spectrometry (SIMS) is a new desorption/ionization technique for mass spectrometry in which highly charged water clusters produced from atmospheric-pressure electrospray are accelerated in vacuum by several kV and impact on the sample deposited on the metal substrate. In this study, we applied EDI/SIMS directly to fruits, such as bananas, strawberries, grapes and apples. The major components in the fruits--fructose, glucose, sucrose and organic acids--could be observed with strong signal intensities. EDI/SIMS was also applied to the analysis of different regions of strawberries and apples. Compared with matrix-assisted laser desorption/ionization (MALDI), ion signals with lower background signals could be obtained, particularly for the low molecular weight analytes.

EFFECT OF BORON AND CALCIUM SPRAYS ON PHOTOSYNTHETIC PIGMENTS, TOTAL PHENOLS AND FLAVONOID CONTENT OF SWEET CHERRY (PRUNUS AVIUM L.)

The aim of our study was to establish the effect of a three-year-long boron and calcium treatment on some phytochemical compounds of sweet cherry (Prunus avium L.) leaves and fruits. In our trial we used eight-year-old cultivar 'Germersdorfi 3' grafted on Prunus mahaleb rootstock. The boron treatment was applied by spraying disodium-octaborat-tetrahydrat, while calcium was supplied as a calcium nitrate spray. Photosynthetic pigments and total phenol content of sweet cherry leaves and total anthocyanin, flavonoid and phenol contents of sweet cherry fruits were measured, respectively. Boron sprays at blooming time had a positive effect on photosynthetic pigment contents of sweet cherry leaves, on contrary to two times spraying, which reduced their content. Total carotenoids responded similarly to boron spraying: single treatment increased their mass, two-times spraying had a negative effect on the total carotenoid content of the leaves, respectively. Every calcium treatment increased the chlorophyll a and chlorophyll b content. The mass of carotenoids were also in close connection with calcium sprays. It is to be noted that time and frequency of application influenced the mass of both photosynthetic pigments. Effect of boron and calcium sprays on total phenol content was significant. After the double boron treatment, quantity of total phenol content was just ninth of the control leaves. Boron and calcium sprays decreased the total phenol, anthocyanin and flavonoids content in sweet cherry fruits, respectively. However, boron treatment was more effective than calcium sprayings. Conclusion of boron and calcium spraying was significant in most cases of our trial.

Protection of Fibroblasts (NIH-3T3) against Oxidative Damage by Cyanidin-3-rhamnoglucoside Isolated from Fig Fruits (Ficus caricaL.)

Anthocyanins, plant secondary metabolites, have been recognized for their health-promoting properties when consumed by humans. In this study, the antioxidant properties of a major anthocyanin in fresh fig fruits, cyanidin-3-rhamnoglucoside (C3R), were evaluated by various assays in vitro and correlated with the protection afforded by C3R to cultured NIH-3T3 fibroblast cells. C3R inhibited lipid peroxidation from producing peroxy radicals (ROO(*)) and MDA in a dose-dependent manner, and a high calculated stoichiometric coefficient [n] for peroxy radicals was demonstrated. In addition to its scavenging of reactive oxygen species (ROS), C3R showed a strong chelating activity toward the Fe(2+) ion. Finally, pretreatment with C3R inhibited proapoptotic processes that were initiated by the oxidation of lysosome membranes in fibroblast cells. The high antioxidant potential, with several modes of action of purified C3R, may contribute to health benefits gained by the consumption of fresh fig fruits.

EPR Studies of O2•−, OH, and 1O2 Scavenging and Prevention of Glutathione Depletion in Fibroblast Cells by Cyanidin-3-rhamnoglucoside Isolated from Fig (Ficus carica L.) Fruits

Cyanidin-3-rhamnoglucoside (C3R) is the major anthocyanin in fresh fig fruits. In this study, the free radical scavenging potential of C3R was evaluated in vitro using several free radical generators. This naturally occurring anthocyanin was superior to other tested natural antioxidants in scavenging ABTS(*+). Electron paramagnetic resonance served to determine the scavenging properties of C3R toward superoxide radical anion (O(2)(*-)), hydroxyl radical ((*)OH), and singlet radical ((1)O(2)). The protection of NIH-3T3 fibroblast cells was then tested as the inhibition of reactive oxygen species (ROS) formation in a dose-dependent manner. It was further demonstrated that treatment with C3R elevates the reduced glutathione (GSH) concentration and the redox ratio (GSH/GSSG) in fibroblast cells in a dose-dependent manner. Moreover, C3R reduced the induction of ROS by butathionine sulfoximine (BSO) and elevated the redox ratio. Thus, it is suggested that C3R in fresh fig fruits is a potent scavenger and may influence endogenous antioxidant systems of consumers.

Analysis and characterisation of anthocyanins in mulberry fruit

The fruit of mulberry (<I>Morus alba</I> L., Moraceae) has been used as medicinal food in China for a long history. The pigment from the fruit extract is a kind of natural colourant for food processing and has potential medical and commercial values. This study focuses on the analysis and characterisation of anthocyanins from mulberry pigment. The fresh mulberry fruits were extracted with the solvent of 95% alcohol/0.1% HC l (1:1, ratio) at room temperature for 4 h in the dark. After the isolation using C-18 column, the pigment was identified with UV-Visible Spectroscopy, HPLC-PAD, LC-MS, and <SUP>1</SUP>HNMR. The results showed that the abundant anthocyanins in mulberry pigment are cyanidin 3-<I>O</I>-rutinoside (60%) and cyanidin 3-<I>O</I>-glucoside (38%). The minor anthocyanins (totally 2%) are pelargonidin 3-<I>O</I>-glucoside and pelargonidin 3-<I>O</I>-rutinoside.

The properties and stability of anthocyanins in mulberry fruits

Mulberry fruit is well known as a good source of anthocyanins with many biological activities. However, there are several colors of mulberry fruits, even from the same species, which may generate different amounts of anthocyanins. This study investigates anthocyanin content and antioxidant levels as well as tyrosinase inhibition activity in the extract from various colors of mulberry fruit, Morus alba. The effects of heat and light, which the extract may be exposed to during food processing, on anthocyanin and antioxidant activities are also evaluated. Our results show that purple-colored mulberry fruit extract contains the highest levels of anthocyanin and strongest antioxidant as well as anti-tyrosinase properties compared with other colors mulberry fruit extracts. Light or heat exposure by incubation of the mulberry fruit extract at 70 °C for 10 h significantly deteriorated total anthocyanin and ascorbic acid content and led to a corresponding increase of the IC50 values.

Electrochemical behaviour and antioxidant capacity of anthocyanins from Chilean red wine, grape and raspberry

abstract The anthocyanin fractions were extracted from Cabernet Sauvignon red wine, skins of Vitis vinifera grapesand raspberry fruits (Rubus idaeus). In red wine extract, 16 anthocyanins were identified, malvidin-3-O-glucoside being the main anthocyanin, which comprised 53.6% of the total anthocyanin in grape extract.Raspberry extract contained mainly delphinidin-3-O-glucoside and cyanidin-3-O-glucoside. The antiox-idant capacity of the extracts was assayed by electrochemical methods. Best resolution of the oxidationpeaks for the extracts and diluted wine was obtained by pulse differential voltammetry. The wine diluted20 presented values of P1 (443 mV) and P2 (676 mV) similar to those corresponding to wine extract,and to the anthocyanin malvidin-3-O-glucoside. The antioxidant capacity of anthocyanins in extractsof wine, grape skin and raspberry fruit was also determined by the Trolox equivalent antioxidant capacity(TEAC) method. 2009 Elsevier Ltd. All rights reserved. 1. IntroductionRed wine is a complex system which contains a large variety ofphenolic compounds, such as flavonols, flavan-3-ols, anthocyaninsand phenolic acids. Anthocyanins are extracted from red grapesand are largely responsible for the colour exhibited by red wine;they play an important role in the antioxidant activity of red wine(Zoecklein, Fugelsang, Gump, & Nury, 2001). In red wine, anthocya-ninsinteractwithphenolicscompounds,pyruvicacidandacetalde-hyde to give new pigments (Francia-Aricha, Guerra, Rivas-Gonzalo,SFulcrand,Bernabdeljalil,Rigaud,Cheynier,M Mateus, Silva, Vercauteren, & De Feitas, 2001).The antioxidative capacity of anthocyanin was studied by differentchemical methods. The Folin–Ciocalteu method, the oxygen radicalabsorbancecapacity(ORAC)assayandtheTroloxequivalentantiox-idantcapacity(TEAC)assayarethemostconsideredmethods(Prior,Wu, & Schaich, 2005). Kilmartin and Hsu (2001) developed anelectrochemical method for the characterization of the antioxidantproperties of wine and wine phenolics. Recently, Janeiro andOliveira Brett (2007) studied the redox behaviour of anthocyaninsby voltammetric techniques, they found that all the phenolichydroxylgroupscanbeelectrochemicallyoxidized.DaSilva,RamosStradiotto,andOliveira(2008)developedamethodforthedetermi-nation of caffeic acid in red wine by differential pulse voltammetry.Zienlin´ska, Nagels, and Piskula (2008) determined the antioxidantcapacity of quercetin and its glycosides of onion by cyclic voltam-metry. Ghiselli, Nardini, Baldi, and Scaccini (1998) reported thatamong the red wine components, anthocyanins were the mosteffectiveinscavengingreactiveoxygenspecies(ROS)andinhibitinglipoprotein oxidation. Anthocyanins comprise around 70% of thetotalphenolsofItalianredwine.Inthisworkweevaluatedtheelec-trochemical behaviour of Chilean Cabernet Sauvignon red wine,Vitis vinifera grape and raspberry anthocyanin extracts in a modelwine solution at pH 3.6, and in acetate buffer.2. Materials and methods2.1. MaterialsCabernet Sauvignon red wine vintage 2005 and V. viniferagrapes were supplied by a local producer. Mature raspberry fruits(Rubus idaeus) were purchased in the local market. All the solventswere reagent grade and/or HPLC grade (Merck, Darmstadt,Germany), all the chemicals were reagent grade (Sigma, St. Louis,MO, USA).2.2. HPLC analysisFor the high performance liquid chromatography (HPLC) analy-sis a Waters 600 HPLC chromatograph (Waters, Mildford, MA, USA)equipped with a Waters 2990 diode array detector, and a

Evidence for a photoprotective function of low‐temperature‐induced anthocyanin accumulation in apple and pear peel

The light requirement and low-temperature stimulation of anthocyanin synthesis in peel of apple (Malus domestica) and pears (Pyrus communis) and the presence of anthocyanins in immature fruits are not congruent with a visual function in dispersal. We hypothesized that anthocyanins afford photoprotection to peel during low-temperature-induced light stress and that the protection is not a fortuitous side-effect of light absorption by anthocyanin. The extent of photoinhibition at harvest and after light stress treatment in pear cultivars differing in redness decreased with increasing red color on the sun-exposed sides of fruits. Green-shaded sides of the pears showed comparable levels of photoinhibition indicating that pears did not differ in their inherent photosensitivity. Apple and pear peel show considerable short-term fluctuation in redness in response to temperature, with red color increasing rapidly in response to low temperature and just as quickly fading in response to high temperature. Briefly, shading pears and apples during cold conditions for 2 days reduced the accumulation of anthocyanin and increased the photosensitivity of peel. Subsequent shading during warm conditions did not affect the accumulation of anthocyanin or the photosensitivity of peel indicating that the response at low temperature was not due to shade adaptation. The assessment of photosystem II (PSII) efficiency and quenching of chlorophyll fluorescence between 16 and 40 degrees C indicated that 'Forelle' pear peel was particularly sensitive to photostress at low temperature. The photosynthetic system in mature 'Forelle' leaves was comparatively much less sensitive to light stress at low temperature. Results support the view that anthocyanins are adaptable light screens deployed to modulate light absorption in sensitive tissues such as fruit peel in response to environmental triggers such as cold front snaps.

Antioxidant properties of blackberry and blueberry fruits grown in the Black Sea Region of Turkey

Seven wild and ten cultivated blackberries (Arapaho, Bartin, Black Satin, Bursa 1, Bursa 2, Cherokee, Chester, Jumbo, Navaho, and Ness), and six lowbush ( Vaccinium arctostaphylos) and four highbush ( Vaccinium corymbosum) blueberries fruits (Ivanhoe, Jersey, Northland, and Rekord) were analyzed for total anthocyanins, total phenolics, and antioxidant activity as ferric reducing antioxidant power (FRAP) in this study. The respective ranges of total anthocyanin and total phenolic contents of the tested samples were: blackberries, 0.95–1.97 and 1.73–3.79 mg g −1 and blueberries, 0.18–2.94 and 0.77–5.42 mg g −1. FRAP values varied from 35.05 to 70.41 μmol g −1 for blackberries, 7.41 to 57.92 μmol g −1 for blueberries. Wild blackberries had the highest FRAP values while wild blueberries had the highest total phenolic and total anthocyanin contents. A linear relationship was observed between FRAP values and total phenolics for blueberries ( r = 0.981). The anthocyanin pigments in samples were isolated and characterized by high-performance liquid chromatography (HPLC) with UV–visible detection. Cyn-3-glu was the predominant anthocyanin in all blackberry fruits.

Influência do pH da solução extrativa no teor de antocianinas em frutos de morango

Nos últimos anos, os estudos sobre a composição química do fruto do morangueiro vêm ganhando destaque devido ao seu elevado consumo e às atividades biológicas atribuídas a ele. Além disso, os principais metabólitos secundários encontrados nesses frutos são as antocianinas, compostos responsáveis pela intensa coloração vermelha e amplamente utilizados como corantes naturais pela indústria alimentícia. Neste trabalho, foram preparados diferentes extratos de frutos de morango com a cultivar Oso Grande, com o intuito de verificar a influência do pH no perfil espectroscópico do extrato e no teor de antocianos. O extrato em pH 1,0 forneceu um perfil clássico para antocianinas na análise por espectroscopia no ultravioleta, assim como um maior teor de antocianinas nos frutos frescos devido a uma extração mais eficiente das substâncias de interesse.

Chemical stability of açai fruit (Euterpe oleracea Mart.) anthocyanins as influenced by naturally occurring and externally added polyphenolic cofactors in model systems

Abstract The influence of different classes of naturally occurring and externally added polyphenolic cofactors on the phytochemical and colour stability of anthocyanins in acai fruit (Euterpe oleracea) was investigated. Model systems were based on anthocyanin isolates from acai fruit, rich in cyanidin-3-rutinoside (311 ± 27 mg/l) and cyanidin-3-glucoside (208 ± 18 mg/l), and isolated groups of naturally occurring polyphenolic cofactors in acai fruit (phenolic acids, procyanidins, and flavone-C-glycosides, each adjusted to ∼50 mg/l). Anthocyanin degradation kinetics were assessed as a function of pH (3.0, 3.5, and 4.0) and storage temperature (5, 20 and 30 °C). During storage, anthocyanins experienced pH and temperature-dependent losses, and the half life cyanidin-3-rutinoside (t1/2 = 2.67–210 days) was consistently longer than cyanidin-3-glucoside (t1/2 = 1.13–144 days). The presence of flavone-C-glycosides induced significant hyperchromic shifts and enhanced anthocyanin stability at all pH and temperature combinations, while no significant effects were attributed to the presence of phenolic acids or procyanidins. Additional models using externally added cofactors from rooibos tea, also rich in flavone-C-glycosides, resulted in up to 45.5% higher anthocyanin colour and up to 40.7% increased anthocyanin stability compared to uncopigmented anthocyanin isolates and had similar copigmentation effects to a commercial rosemary-based colour enhancer. Results suggest flavone-C-glycosides offer potential for their use as colour enhancers and stabilizing agents in products rich in cyanidin glycosides, particularly acai fruit-containing foods, juice blends, and beverages.

Structure of Anthocyanins from Eugenia Jambolana Fruit

The purple color of the ripe Eugenia jambolana fruit is attributed to anthocyanins, a class of plant pigments that has attracted immense attention due to the potential health benefits of these compounds. There is disagreement in the literature on whether E. jambolana fruit anthocyanins are found as 3,5- or 3-diglucosides. Therefore, we used a combination of HPLC-UV, tandem LC-MS, and NMR techniques to identify the structures of anthocyanins present in E. jambolana fruit collected in the U.S.A. and India. Our results indicate that the anthocyanins from both locations occur as 3,5-, but not 3-diglucosides, of delphinidin, cyanidin, petunidin, peonidin, and malvidin. This is the first report to use a combination of spectrometric and spectroscopic methods to identify unequivocally the structures of E. jambolana fruit anthocyanins.

European elderberry ( Sambucus nigra L.) rich in sugars, organic acids, anthocyanins and selected polyphenols

Sugars and organic acids in the fruit of two cultivars and three selections of black elderberry ( Sambucus nigra L.): ‘Haschberg’, ‘Rubini’, ‘Selection 13’, ‘Selection 14’ and ‘Selection 25’ were quantified. The anthocyanin as well as quercetin profiles of this plant material were also established by the use of HPLC/MS. Significant differences in the concentration of sugars and organic acids were detected between the widely spread cultivar ‘Haschberg’ and all other cultivars/selections; ‘Haschberg’ was the richest in organic acids (6.38 g kg −1 FW), and it contained the least sugar (68.5 g kg −1 FW). The following major cyanidin based anthocyanins were identified in the fruit of black elderberry: cyanidin 3-sambubioside-5-glucoside, cyanidin 3,5-diglucoside, cyanidin 3-sambubioside, cyanidin 3-glucoside and cyanidin 3-rutinoside. The most abundant anthocyanin in elderberry fruit was cyanidin 3-sambubioside, which accounted for more than half of all anthocyanins identified in the berries. The ‘Rubini’ cultivar had the highest amount of the anthocyanins identified (1265 mg/100 g FW) and the lowest amount was measured in berries of the ‘Selection 14’ (603 mg/100 g FW). The ‘Haschberg’ cultivar contained a relatively low amount of anthocyanins in ripe berries (737 mg/100 g FW). From the quercetin group, quercetin, quercetin 3-rutinoside and quercetin 3-glucoside were identified; the latter prevailing in black elderberry fruit. The cultivar with the highest amount of total quercetins was ‘Selection 25’ (73.4 mg/100 g FW), while the ‘Haschberg’ cultivar contained average amounts of quercetins (61.3 mg/100 g FW). The chemical composition of the ‘Haschberg’ cultivar, the most commonly planted, conforms to the standards for sugars, anthocyanins and quercetins and exceeds them in the content levels of organic acids, the most important parameter in fruit processing.

Anthocyanins Reduce Fungal Growth in Fruits

As the most visible role of anthocyanins is to impart colors, the adaptive significance of anthocyanins in plant reproductive organs is invariably attributed to the attraction of seed dispersers and pollinators. However, few studies suggested that anthocyanins in vegetative tissue might also function in plant defence, although the evidence for such a function is not particularly strong. Here, we investigated whether anthocyanins contribute to fruit defence against fruit-rot fungi. We found that the risk of fruit-rot in grape varieties infected with Botrytis cinerea decreased with increasing anthocyanin contents. Moreover, anthocyanin contents directly inhibited growth rates of nine fruit-rot fungi on agar plates. Anthocyanins reduced fungal growth by 50% in the concentrations that typically characterise unripe blackberries and by 95% in the concentrations that typify ripe blackberries. We conclude that anthocyanins in fruits not only function to attract animal vectors, but that they also contribute to fruit defence. The antifungal activity of anthocyanins might more widely explain their occurrence in various plant organs such as vegetative tissue and roots.

Total content of phenols and anthocyanins in fruits from Bosnia

Total content of phenols and anthocyanins in fruits from Bosnia

MYB transcription factors that colour our fruit

Anthocyanin concentration is a primary determinant of plant colour. Fruit anthocyanin biosynthesis is controlled by a distinct clade of R2R3 MYB transcription factors. In apple, three recent papers describe the discovery of MYB genes activating skin, flesh and foliage anthocyanic colour. These findings lead the way to new approaches in the breeding and biotechnological development of fruit with new colour patterns.

Molecular Aspects of Anthocyanin fruit Tomato in Relation to high pigment-1

The tomato Anthocyanin fruit (Aft) genotype is characterized by purple color in skin and outer pericarp of its fruits due to higher levels of anthocyanins-flavonoid metabolites. Our objectives were to carry out metabolic and molecular characterization of this genotype, emphasizing its interaction with the high pigment-1 (hp-1) mutation, known to increase flavonoids in tomato fruits. These objectives fit the growing interest in developing tomato fruits with higher levels of functional metabolites. Our results show that 1) Aft fruits are also characterized by significantly higher levels of the flavonols quercetin and kaempferol, thus enhancing their functional value; 2) the tomato Anthocyanin1 (Ant1) gene, encoding a Myb transcription factor, displayed nucleotide and amino acid polymorphisms between the Aft genotype and cultivated genotypes; 3) a DNA marker based on Ant1 showed that the Aft trait is encoded by a single locus on chromosome 10 fully associated with Ant1; and 4) double homozygotes Aft/Aft hp-1/hp-1 plants displayed a more-than-additive effect on the production of fruit anthocyanidins and flavonols. This effect was manifested by approximately 5-, 19-, and 33-fold increase of petunidin, malvidin, and delphinidin, respectively, in the double mutants compared with the cumulative levels of their parental lines.