Anthocyanin composition of Brassica oleracea cv. Red Danish

Eight anthocyanins were isolated from illuminated red cabbage seedlings. They were identified as: cyanidin-3-sophoroside-5-glucoside, cyanidin-3-malonyl-sophoroside-5-glucoside, cyanidin-3-p-coumaryl-sophoroside-5-glucoside, cyanidin-3-(di-p-coumaryl)sophoroside-5-glucoside, cyanidin-3-ferulyl-sophoroside-5-glucoside, cyanidin-3-(diferulyl)sophoroside-5-glucoside, cyanidin-3-sinapylsophoroside-5-glucoside, and cyanidin-3-(disinapyl)-sophoroside-5-glucoside.     

A rationale for the shift in colour towards blue in transgenic carnation flowers expressing the flavonoid 3',5'-hydroxylase gene

Recently marketed genetically modified violet carnations cv. Moondust and Moonshadow (Dianthus caryophyllus) produce a delphinidin type anthocyanin that native carnations cannot produce and this was achieved by heterologous flavonoid 3',5'-hydroxylase gene expression. Since wild type carnations lack a flavonoid 3',5'-hydroxylase gene, they cannot produce delphinidin, and instead accumulate pelargonidin or cyanidin type anthocyanins, such as pelargonidin or cyanidin 3,5-diglucoside-6"-O-4, 6"'-O-1-cyclic-malyl diester. On the other hand, the anthocyanins in the transgenic flowers were revealed to be delphinidin 3,5-diglucoside-6"-O-4, 6"'-O-1-cyclic-malyl diester (main pigment), delphinidin 3,5-diglucoside-6"-malyl ester, and delphinidin 3,5-diglucoside-6",6"'- dimalyl ester. These are delphinidin derivatives analogous to the natural carnation anthocyanins. This observation indicates that carnation anthocyanin biosynthetic enzymes are versatile enough to modify delphinidin. Additionally, the petals contained flavonol and flavone glycosides. Three of them were identified by spectroscopic methods to be kaempferol 3-(6"'-rhamnosyl-2"'-glucosyl-glucoside), kaempferol 3-(6"'-rhamnosyl-2"'-(6-malyl-glucosyl)-glucoside), and apigenin 6-C-glucosyl-7-O-glucoside-6"'-malyl ester. Among these flavonoids, the apigenin derivative exhibited the strongest co-pigment effect. When two equivalents of the apigenin derivative were added to 1 mM of the main pigment (delphinidin 3,5-diglucoside-6"-O-4,6"'-O-1-cyclic-malyl diester) dissolved in pH 5.0 buffer solution, the lambda(max) shifted to a wavelength 28 nm longer. The vacuolar pH of the Moonshadow flower was estimated to be around 5.5 by measuring the pH of petal. We conclude that the following reasons account for the bluish hue of the transgenic carnation flowers: (1). accumulation of the delphinidin type anthocyanins as a result of flavonoid 3',5'-hydroxylase gene expression, (2). the presence of the flavone derivative strong co-pigment, and (3). an estimated relatively high vacuolar pH of 5.5.     

Covalent anthocyanin-flavonol complexes from flowers of chive, Allium schoenoprasum

The structures of eight anthocyanins have been determined in acidified methanolic extract of pale-purple flowers of chive, Allium schoenoprasum. Four of them have been identified as the anthocyanin-flavonol complexes (cyanidin 3-O-beta-glucosideAII) (kaempferol 3-O-(2-O-beta-glucosylFIII-beta-glucosideFII)-7-O-beta-gl ucosiduronic acidFIV) malonateAIII (AII-6-->AIII-1, FIV-2-->AIII-3), 1, (cyanidin 3-O-(3-O-acetyl-beta-glucosideAII) (kaempferol 3-O-(2-O-beta-glucosylFIII-beta-glucosideFII)-7-O-beta-gl ucosiduronic acidFIV) malonateAIII (AII-6-->AIII-1, FIV-2-->AIII-3), 2, and their 7-O-(methyl-O-beta-glucosiduronateFIV) analogous, 3 and 4. Pigments 1 and 2 are the first final identification of covalent complexes between an anthocyanin and a flavonol, while 3 and 4 are formed during the isolation process. The other four anthocyanins (5-8) were found to be the 3-acetylglucoside, 3-glucoside, 3-(6-malonylglucoside) and 3-(3,6-dimalonylglucoside) of cyanidin. The three latter pigments have earlier been identified as the major anthocyanins of the chive stem. The covalent anthocyanin-flavonol complexes show intramolecular association between the anthocyanidin (cyanidin) and flavonol (kaempferol) units, which influence the colour.     

Anthocyanins of Cephaelis, Cynomorium, Euterpe, Lavatera and Pinanga

The two malonylated pigments, malonylmalvin and malvidin 3-malonylglucoside, were identified in petals of Lavatera maritima, which belongs to the Malvaceae, a family known to synthesise such pigments. Zwitterionic anthocyanins could not be detected in four other newly examined sources and common unacylated pigments were recorded. Thus, the fruits of the palms Euterpe edulis and Pinanga polymorpha have a mixture of cyanidin 3-glucoside and cyanidin 3-rutinoside, while the fruit of Cephaelis subcoriacea is coloured by cyanidin 3-glucoside. The latter pigment was also obtained from the reddish brown inflorescence of the parasitic plant Cynomorium coccineum.     

Malonylated Anthocyanins from Bulbs of Red Onion,Allium cepa L.

Four cyanidin-based anthocyanins (1–4) were isolated from the red onion, Allium cepa L. Pigments 1 and 3 were identified as cyanidin 3-glucoside (Cy 3-Glc) and 3-malonylglucoside (Cy 3-MaGlc), respectively, by cochromatography with standard pigments. Anthocyanins 2 and 4 were respectively determined as cyanidin 3-laminaribioside (Cy 3-Lam) and 3-malonyllaminaribioside (Cy 3-MaLam), a new anthocyanin, mainly by NMR tech-niques. Malonylated anthocyanins 3 and 4 were found for the first time in red onions.     

Anthocyanins of grasses

The anthocyanin content of 23 grass species (Poaceae) belonging to five subfamilies has been determined. Altogether 11 anthocyanins were identified; the 3-(6″-malonylglucosides) and 3-glucosides of cyanidin, peonidin and delphinidin, the 3-(3″,6″-dimalonylglucoside), 3-(6″-rhamnosylglucoside) and 3-(6″-glucosylglucoside) of cyanidin, in addition to peonidin 3-(dimalonylglucoside) and delphinidin 3-(6″-rhamnosylglucoside). Anthocyanins acylated with one and/or two malonic acid moieties dominated the anthocyanin profiles of all the species in the subfamilies Pooideae and Panicoideae. On the other hand, the 3-glucoside and 3-rutinoside of cyanidin were the major anthocyanins in Sinarundinaria murielae (subfamily Bambusoideae) and Molinia caerulea (subfamily Arundinoideae), while the 3-glucosides of cyanidin and peonidin were the principal anthocyanins in rice, Oryza sativum (subfamily Oryzoideae). Pelargonidin derivatives and free anthocyanidins have previously been reported to occur in several Poaceae species, however, not identified in any of the species included in this survey.     

Anthocyanins in the epacridaceae

An examination of 73 species of the family Epacridaceae resulted in the identification of the following anthocyanins: cyanidin 3-galactoside, cyanidin 3-glucoside, cyanidin 3-arabinoside, cyanidin 3-rhamnoside, cyanidin 3-rhamnosylgalactoside, cyanidin 3-rhamnosylglucoside, cyanidin 3-xylosylgalactoside, cyanidin 3-xylosylarabinoside, delphinidin 3-galactoside, delphinidin 3-arabinoside, delphinidin 3-rhamnosylgalactoside, delphinidin 3-rhamnosylglucoside and pelargonidin 3-rhamnosylglucoside. No acylated or 5-substituted anthocyanins were detected in any of the species examined. Evidence of methylated anthocyanidin was found only in one species, Woollsia pungens. The occurrence of cyanidin 3-galactoside and cyanidin 3-arabinoside forms a chemical link between this family and the related Ericaceae.     

Acylated anthocyanins and flavonols from purple flowers of Dendrobium cv. ‘Pompadour’

Two rare anthocyanins, cyanidin 3-(6-malonylglucoside)-7,3′-di(6-sinapylglucoside) and the demalonyl derivative, were characterised as the purple floral pigments of Dendrobium cv. ‘Pompadour’. Nine known flavonol glycosides were also identified, including the 3-rutinoside-7-glucosides of kaempferol and quercetin. One new glycoside was detected: the ferulyl ester of quercetin 7-rutinoside-7-glucoside. These flavonoid patterns are typical for plants in the family Orchidaceae.     

Anthocyanins from red flower tea (Benibana-cha), Camellia sinensis

Three anthocyanins were isolated from the leaves of red flower tea (Benibana-cha), Camellia sinensis, and their structures were determined by means of chemical and spectroscopic analyses. Two are the anthocyanins, delphinidin and cyanidin 3-O-beta-D-galactosides, respectively. Whereas the third, delphinidin 3-0-beta-D-(6-(E)-p-coumaryl)galactopyranoside. The anthocyanins were also contained in the flowers of Benibana-cha in different compositions.     

Distribution of anthocyanins in aceraceae leaves

The distribution of anthocyanins in spring sprouted and/or autumn coloured leaves of Dipteronia sinensis and Acer (119 taxa) was studied.

Dipteronia contained four cyanidin glycosides: the 3-glucoside, 3-rutinoside, 3-galloylglucoside and 3,5-diglucoside. Acer contained five cyanidin glycosides: 3-glucoside, 3-rutinoside, 3-galloylglucoside, 3-galloylrutinoside and 3,5-diglucoside, two delphinidin glucosides: 3-glucoside and 3-rutinoside and three unidentified anthocyanins. Both Dipteronia and Acer contained the recently reported cyanidin 3-galloylglucoside. The anthocyanin constituents in spring leaves were more complex than those found in autumn coloured leaves: nine in spring and six in autumn. The presence/absence of the major anthocyanins in the spring sprouted leaves of 111 Acer taxa analysed were grouped into 17 distribution patterns. In the autumn the number of anthocyanin distribution patterns was found to be 11. In Acer, cyanidin glycosides were found in 20 sections and delphinidin glycosides in 17 out of the 21 sections analysed. Although the distribution of anthocyanins showed no clear relations among sections, delphinidin glycosides were mainly found in sections Macrantha, Goniocarpa and Saccharina. There were no differences in the pigment constituents in the species native to different countries, such as A. rubrum in North America and A. pycnanthum in Japan, both containing the same pigments: cyanidin 3-glucoside, 3-rutinoside, 3-galloylglucoside, 3-galloylrutinoside and 3,5-diglucoside.     

Uridine 5′-diphosphate-xylose: anthocyanidin 3-O-glucose-xylosyltransferase from petals of Matthiola incana R.Br.

Petals of genetically defined lines of Matthiola incana R.Br. contain a glycosyltransferase which catalyzes the transfer of the xylosyl moiety of uridine 5-diphosphate-xylose to the glucose of cyanidin 3-glucoside. The enzyme also uses 3-glucosides of pelargonidin and delphinidin, cyanidin 3-(p-coumaroyl)-glucoside and 3-(caffeoyl)-glucoside as substrates. The xylosyltransferase exhibits a pH optimum of 6.5. The enzyme activity depends on the stage of bud and flower development. Accumulation of cyanidin 3-glucoside during flower development is correlated with xylosyltransferase activity.Abbreviations HPLC high-performance liquid chromatography - UDP uridine 5-diphosphate     

Differences in the floral anthocyanin content of red petunias and Petunia exserta

In order to resolve a conflict between previous papers regarding the floral anthocyanins of red flowers of Petunia exserta, a naturally occurring species, the HPLC profile of this species was compared with that of commercial red garden petunias. Both HPLC profiles extremely superficially resemble each other in terms of relative amounts and retention times of the major anthocyanins. However, co-elution on HPLC of the mixed sample resulted in clear separation of the components. Three major anthocyanins in red petunias were determined to be cyanidin 3-sophoroside, cyanidin 3-glucoside and peonidin 3-glucoside, which exhibited similar behaviors on HPLC to delphinidin 3-glucoside. delphinidin-3-rutinoside and petunidin 3-rutinoside, respectively, the major floral anthocyanins of P. exserta.     

Anthocyanins in berries of Maqui (Aristotelia chilensis (Mol.) Stuntz)

The anthocyanin composition of berries of Maqui [Aristotelia chilensis (Mol.) Stuntz] was determined by HPLC with photodiode array and MS detection. Eight pigments corresponding to the 3-glucosides, 3,5-diglucosides, 3-sambubiosides and 3-sambubioside-5-glucosides of delphinidin and cyanidin were identified, the principal anthocyanin being delphinidin 3-sambubioside-5-glucoside (34% of total anthocyanins). The average total anthocyanin content was 137.6 +/- 0.4mg/100g of fresh fruit (211.9 +/- 0.6 mg/100g of dry fruit). The relative high anthocyanin content and the important presence of polar polyglycosylated derivatives makes the fruits of A. chilensis an interesting source of anthocyanin extracts for food and pharmaceutical uses.     

Correlations between anthocyanin chemistry and pollination ecology in the polemoniaceae

A study of the anthocyanins in a representative sample (34 species from 14 genera) of Polemoniaceae has shown that the pigment type in the flowers is broadly correlated with pollination ecology. Thus, hummingbird pollinated species such as Ipomopsis aggregata generally contain pelargonidin sometimes with cyanidin, while bee and beefly pollinated species (e.g. Gilia latiflora) contain mainly delphinidin. On the other hand, lepidopteran species such as Leptodactylon californicum have cyanidin or mixtures of cyanidin with delphinidin. The above three anthocyanidins occur usually as the 3-glucoside, 3,5-diglucoside, $\text{3-(p-}\text{coumarylglucoside}\text{)}$ and $\text{3-(p-}\text{coumarylglucoside}\text{)-5-}\text{glucoside}$, although other types are occasionally found. The distribution of glycosidic types and of acylation, unlike that of the anthocyanidins, is more closely correlated with systematic position than with pollinating vectors. In autogamous species where animal pollination is absent or unimportant, anthocyanin pigmentation in the flowers retains the complexity present in related animal-pollinated taxa. Anthocyanins were also identified in hummingbird pollinated plants from two related families and pelargonidin derivatives were detected. In Fouquieria splendens (Fouquieriaceae), the glycosidic pattern was different from that in Polemoniaceae in being 3-galactoside. In Penstemon (Scrophulariaceae) a study of flower anthocyanins was consistent with Straw's hypothesis that the wasp-pollinated P. spectabilis originated by hybridization between the hummingbird-pollinated P. centranthifolius and the bee-pollinated P. grinnellii.     

Cyanidin 3-[6-(p-coumaroyl)-2-(xylosyl)-glucoside]-5-glucoside and other anthocyanins from fruits of Sambucus canadensis.

From the fruits of Sambucus canadensis four anthocyanin glycosides have been isolated by successive application of an ion-exchange resin, droplet-counter chromatography and gel filtration. The structure of the novel, major (69.8%) pigment, cyanidin 3-O-[6-O-(E-p-coumaroyl-2-O-(beta-D-xylopyranosyl)-beta-D- glucopyranoside]-5-O-beta-D-glucopyranoside, was determined by means of chemical degradation, chromatography and spectroscopy, especially homo- and heteronuclear two-dimensional NMR techniques. The other anthocyanins were identified as cyanidin 3-sambubioside-5-glucoside (22.7%), cyanidin 3-sambubioside (2.3%) and cyanidin 3-glucoside (2.1%).     

Isolation and structural determination of 13 flavonoid glycosides in Hibiscus esculentus (okra)

Eleven flavonol glycosides and two anthocyanins, only one of which was previously identified, were isolated from the flower petals of okra, Hibiscus esculentus L. On the basis of chromatographic, spectral, and degradative evidence, the following structural assignments were made: quercetin 4′-glucoside, quercetin 7-glucoside, quercetin 5-glucoside, quercetin 3-diglucoside, quercetin 4′-diglucoside, quercetin 3-triglucoside, quercetin 5-rhamnoglucoside, gossypetin 8-glucoside, gossypetin 8-rhamnoglucoside, gossypetin 3-glucosido-8-rhamnoglucoside, cyanidin 4′-glucoside, and cyanidin 3-glucosido-4′ glucoside. Some evidence was obtained of a pentahy-droxy, monomethoxy-flavone glycoside. The total flavonoid content in the red portion of the petal was 0.48% of fresh weight; that in the white portion was 2.51%. The two anthocyanins comprised 28.5% of the flavonoid content of the red flower but only a trace of the content of the white.     

Relationship between the composition of flavonoids and flower colors variation in tropical water lily (Nymphaea) cultivars

Water lily, the member of the Nymphaeaceae family, is the symbol of Buddhism and Brahmanism in India. Despite its limited researches on flower color variations and formation mechanism, water lily has background of blue flowers and displays an exceptionally wide diversity of flower colors from purple, red, blue to yellow, in nature. In this study, 34 flavonoids were identified among 35 tropical cultivars by high-performance liquid chromatography (HPLC) with photodiode array detection (DAD) and electrospray ionization mass spectrometry (ESI-MS). Among them, four anthocyanins: delphinidin 3-O-rhamnosyl-5-O-galactoside (Dp3Rh5Ga), delphinidin 3-O-(2"-O-galloyl-6"-O-oxalyl-rhamnoside) (Dp3galloyl-oxalylRh), delphinidin 3-O-(6"-O-acetyl-β-glucopyranoside) (Dp3acetylG) and cyanidin 3- O-(2"-O-galloyl-galactopyranoside)-5-O-rhamnoside (Cy3galloylGa5Rh), one chalcone: chalcononaringenin 2'-O-galactoside (Chal2'Ga) and twelve flavonols: myricetin 7-O-rhamnosyl-(1 → 2)-rhamnoside (My7RhRh), quercetin 7-O-galactosyl-(1 → 2)-rhamnoside (Qu7GaRh), quercetin 7-O-galactoside (Qu7Ga), kaempferol 7-O-galactosyl-(1 → 2)-rhamnoside (Km7GaRh), myricetin 3-O-galactoside (My3Ga), kaempferol 7-O-galloylgalactosyl-(1 → 2)-rhamnoside (Km7galloylGaRh), myricetin 3-O-galloylrhamnoside (My3galloylRh), kaempferol 3-O-galactoside (Km3Ga), isorhamnetin 7-O-galactoside (Is7Ga), isorhamnetin 7-O-xyloside (Is7Xy), kaempferol 3-O-(3"-acetylrhamnoside) (Km3-3"acetylRh) and quercetin 3-O-acetylgalactoside (Qu3acetylGa) were identified in the petals of tropic water lily for the first time. Meanwhile a multivariate analysis was used to explore the relationship between pigments and flower color. By comparing, the cultivars which were detected delphinidin 3-galactoside (Dp3Ga) presented amaranth, and detected delphinidin 3'-galactoside (Dp3'Ga) presented blue. However, the derivatives of delphinidin and cyanidin were more complicated in red group. No anthocyanins were detected within white and yellow group. At the same time a possible flavonoid biosynthesis pathway of tropical water lily was presumed putatively. These studies will help to elucidate the evolution mechanism on the formation of flower colors and provide theoretical basis for outcross breeding and developing health care products from this plant.     

Anthocyanins and flavonols from the blue flowers of six Meconopsis species in Bhutan

Blue flowers of six Bhutani Meconopsis species, M. bhutanica, M. bella, M. horridula, M. simplicifolia, M. primulina and M. polygonoides, were surveyed for anthocyanins and other flavonoids. Four anthocyanins were isolated and identified as cyanidin 3-O-sambubioside-7-O-glucoside (1), cyanidin 3-O-[xylosyl-(1 → 2)-(6″-malonylglucoside)]-7-O-glucoside (2), cyanidin 3-O-sambubioside (4) and cyanidin 3-O-[xylosyl-(1 → 2)-(6″-malonylglucoside)] (5). On the other hand, 12 flavonols were isolated from their Meconopsis species with various combination and characterized as kaempferol 3-O-glycosides (8–12), kaempferol 3,7-O-glycosides (13–16), quercetin 3-O-glycosides (17 and 18) and isorhamnetin 3-O-glycoside (19). Of six Meconopsis species which were surveyed in this experiment, anthocyanin and flavonol composition of five species except for M. horridula was clarified for the first time. Their Meconopsis species showed the different flavonoid profiles, respectively, and flavonoid diversity within the glycosylation level of Meconopsis flowers were indicated.     

The identification of flavonoids and the expression of genes of anthocyanin biosynthesis in the chrysanthemum flowers

In order to provide additional information on the coloration of chrysanthemum flowers, the flavonoid composition and the expression of six structural genes involved in anthocyanin pathway in the ray florets of a pink flowering (cv. H5) and two white flowering (cvs. Keikai and Jinba) Chrysanthemum grandiflorum cultivars were examined. HPLCDAD/ESI-MSⁿ analysis showed that cyanidin 3-O-(6″-O-malonylglucoside) and cyanidin 3-O-(3″,6″-O-dimalonylglucoside) were the two major flavonoids presented in H5, while white flowering cultivars contained flavones instead of anthocyanins. Nine flavone derivatives were detected in the three cultivars, the amount of each flavone varied upon cultivars, and seven of these were identified as luteolin 7-O-arabinosylglucuronide, apigenin 7-O-glucoside, luteolin 7-O-malonylglucoside, apigenin 7-O-malonylglucoside, chrysoeriol 7-O-malonylglucoside, acacetin 7-O-rutinoside and acacetin 7-O-malonylglucoside. The two white flowering cultivars showed similar total flavonoid content, which was about two fold higher than that in H5. A high expression of the genes encoding dihydroflavonol 4-reductase and 3-O-glucosyltransferase was detected only in H5 but not in Keikai or Jinba. Chalcone synthase, chalcone isomerase, flavanone 3-hydroxylase, and flavonoid 3′-hydroxylase were expressed in all flowers, suggesting that the lack of anthocyanin in white flowering cultivars cannot be due to any blockage of their expression.     

A survey of anthocyanins in fruits of some angiosperms,I

The anthocyanin pigments in the fruits of fifty-two species belonging to seventeen families of angiosperms were investigated paper-chromatographicallly. They were identified as cyanidin 3-monoglucoside, pelargonidin 3-monoglucoside, cyanidin 3-rutinoside, pelargonidin 3-rutinoside, cyanidin 3-xylosylglucoside, cyanidin 3-xylosylgalactoside, delphinidin 3-xylosylglucoside and delphinidin 3-sophorosido-5-monoglucoside. Of those anthocyanins detected, the most common was cyanidin 3-monoglucoside. In general, the plants belonging to a certain genus contained the same anthocyanin.