Proanthocyanidins and potential precursors in needles of douglas fir and in cell suspension cultures derived from seedling shoot tissues

Proanthocyanidins and their potential precursors have been analyzed by paper chromatography and C₁₈ reversed phase columns with high performance liquid chromatography. Total proanthocyanidins on a dry weight basis in cell suspension cultures derived from seedlings of Douglas fir (Pseudotsuga menziesii) were equal to or greater than those found in mature needles of randomly selected outdoor-grown trees. The major monomer and dimer were catechin and epicatechin-catechin, respectively. Although only procyanidins were detectable in cell suspension cultures, mature needles of outdoor-grown trees contained prodelphinidins as well. Immature needles (flush growth) of the same trees contained only trace amounts of prodelphinidins. Eriodictyol-7-glucoside and dihydroquercetin-3′-glucoside were present in all tissues examined. The amount of eriodictyol-7-glucoside was strongly correlated with total proanthocyanidins in immature needles of flush growth (r = 0.89, p = 0.001). The most complex pattern of flavonoids was found in flush growth needles, which contained in addition to the above, naringenin-7-glucoside and five to six flavone glycosides. Chlorogenic acid was detected only in seedlings and in flush growth needles.     

The flavonoids of phragmites australis flowers

The major flavonoid constituents of Phragmites australis flowers are the C-glycosylflavones swertiajaponin, isoswertiajaponin and two new O-glycosides, the 3′-O-gentiobioside and the 3′-O-glucoside of swertiajaponin. Two unusual flavonol glycosides, rhamnetin 3-O-rutinoside and rhamnetin 3-O-glucoside, were also characterized from the same tissue.     

Cyanidin glycosides in flowers of genus Corydalis (Fumariaceae)

Nine taxa of Corydalis were surveyed for their floral anthocyanins. Five cyanidin glycosides: cyanidin 3-glucoside, cyanidin 3-sambubioside, cyanidin 3-rutinoside, cyanidin 3-(2^G-xylosylrutinoside) and cyanidin 3-(2^G-xylosylrutinoside)-7-glucoside were isolated from these taxa and identified by chemical and spectroscopic techniques. A novel anthocyanin was found in the flowers of Corydalis elata and Corydalis flexuosa cultivars, and identified to be cyanidin 3-(2^G-xylosylrutinoside)-7-glucoside. Two anthocyanins, cyanidin 3-sambubioside and cyanidin 3-(2^G-xylosylrutinoside), were also found for the first time in Corydalis flowers. Furthermore, the major anthocyanin constituent of the flowers was cyanidin 3-sambubioside in the outer petals of Corydalis ambigua and Corydalis lineariloba, and cyanidin 3-rutinoside in those of Corydalis decumbens, Corydalis curvicalcarata and Corydalis speciosa. Similarly, Corydalis incisa contained cyanidin 3-(2^G-xylosylrutinoside), and C. flexuosa ‘China Blue’ and ‘Blue Panda’, and C. elata contained the most complex structural pigment, cyanidin 3-(2^G-xylosylrutinoside)-7-glucoside, as their dominant anthocyanin in their outer petals. According to the results of anthocyanin analyses, these nine plants were classified into four groups: groups A (three taxa), B (two taxa), C (one taxa) and D (three taxa). On the other hand, the anthocyanin constituent of their inner petals was composed of cyanidin 3-rutinoside as only one dominant anthocyanin.     

Iridoid glucosides in Plantago alpina and P. Altissima

Two species of Plantago, namely P. Alpina and P. altissima were investigated. From the former, nine iridoid glucosides and verbascoside were isolated. Together with the known iridoids gardoside, geniposidic acid, 8-epi-loganic acid, mussaenosidic acid, aucubin, monomelittoside and melittoside, two new glucosides were found: 10-O-acetylgeniposidic acid and alpinoside, another compound with a 10-O-acetyl group. From P. altissima verbascoside and isoverbascoside were isolated together with the known iridoids gardoside, 8-epi-loganic acid, catalpol, aucubin, and hookerioside as well as the new compound desacetylhookerioside.     

Trans-fused Iridoid GlycosidEs from Penstemon Mucronatus

Two new trans-fused iridoid glycosides (5αH)-6α-8-epidihydrocornin and (5αH)-6α-8-hydroxy-8-epiloganin, were isolated from Penstemon mucronatus, along with cornin, penstemoside and three hastatosides. The trans-fused iridoids are only the second and third known among over 900 described cis-fused iridoid glycosides. Two pairs of iridoids, identical except for the stereochemistry at C-8, were found. Structures were determined by spectroscopic methods.     

Flavonoids of someVigna-plants in leguminosae

Flavonoids in five lines ofVigna mungo, fifteen lines ofV. radiata var.radiata and two lines ofV. radiata var.sublobata, which belong to the subgenus Ceratotropis, were examined. In hypocotyls, seed-coats and mature leaves of those plants, twelve kinds of flavonoid including three anthocyanins, two leucoanthocyanins, two glycoflavones and five flavonol glycosides were found, and their distribution pattern in twenty-two lines of the legumes is discussed. The leaves ofV. radiata var.radiata and var.sublobata contained the glycosides (mainly rutin) of both quercetin and kaempferol, while those ofV. mungo contained only kaempferol glycosides, with robinin as the predominant pigment, and the purple-red hypocotyls of the former group contained delphinidin 3-p-coumaroylglucoside, while those of the latter contained cyanidin 3-glucoside, although delphinidin 3-glucoside was commonly found in all of the plants. With the exception of two lines, all of the seed-coats examined contained in common four compounds of vitexin, isovitexin, leucocyanidin and leucodelphinidin, whereas in addition to these pigments the black seed-coats ofV. mungo andV. radiata var.sublobata TC 1965 contained delphindin 3-glucoside.     

The Structures of Two Flavonoid Glycoside from Oxytropis ochrocephala

Two flavonoid glycosides separated from water soluble fraction of Oxytropis ochrocephala Bge. were identified by using IR, NMR, and MS spectroscopic methods. The aglycones and sugars of the two compounds were determined directly from the hydrolytic solutions by using a sugar column and a bonded phase column. The results showed that compound 1 contained two kinds of sugars which have the same retention times as galactose and glucose, compound 2 only contained galactose. The aglycones of the two compounds have the same retention times and UV-spectra of rhamnocitrin. Finally, the structures of the two compounds were elucidated as rhamnocitrin-3-galactoside-4’-glucoside and rhamnocitrin-3-galactoside. The first one is a new flavonoid glycoside and the second one is, for the first time, found in the Oxytropis species.     

Flavonoid glycosides and the chemosystematics of Eucalyptus camaldulensisis

Leaf flavonoid glycosides of Eucalyptus camaldulensis were identified as kaempferol 3-glucoside and 3-glucuronide; quercetin 3-glucoside, 3-glucuronide, 3-rhamnoside, 3-rutinoside and 7-glucoside, apigenin 7-glucuronide and luteolin 7-glucoside and 7-glucuronide. Two chemical races were observed based on the flavonoid glycosides. These races correspond to the northern and southern populations of species growing in Australia. The Middle Eastern species examined were found to belong to the southern Australian chemical race. The major glycosides of E. occidentalis proved to be quercetin and myricetin 3-glucuronide.     

An evaluation of the role of cytokinins in the development of abnormal inflorescences in oil palms (Elaeis guineensis Jacq.) regenerated from tissue culture

Tissue cultures and regenerant plants from cell lines producing palms with normal and abnormal flowers were analyzed for cytokinin content and compared with zygotic embryos and seedlings. Immature inflorescences at the critical stage of flower development dissected from normal and abnormal palms were also analyzed. High performance liquid chromatography (HPLC)/radioimmunoassay and HPLC/enzyme-linked immunosorbent assay methods were used over a period of several years to measure the isoprenoid cytokinins. The results of analyses of endogenous aromatic cytokinins, present at comparable levels, will be reported separately. Oil palm cultures and regenerant plants contained relatively high concentrations of the 9-glucosides of isopentenyladenine ([9G]iP) and zeatin ([9G]Z). The predominant biologically active isoprenoid cytokinin present was zeatin riboside ([9R]Z), with lesser amounts of isopentenyladenine (iP) and isopentenyladenosine ([9R]iP). There was evidence of small amounts of dihydrozeatin compounds, but high concentrations (mainly as dihydrozeatin-9-glucoside ([9G]DHZ)) were confined to the haustorium of the zygotic embryo. Callus tissue contained very low concentrations of cytokinin. Frequently only [9G]iP could be detected, at about 1 pmol · g^-1 fresh weight, with [9R]Z at less than 0.05 pmol · g^-1. In comparison, nodular embryogenic tissues in vitro contained between 30 and 1,500 pmol · g^-1 of [9G]iP, 5–50 pmol · g^-1 of [9G]Z, and up to 12 pmol · g^-1 of [9R]Z. Shoots of regenerant plantlets and seedlings contained lower concentrations of [9G]iP (3–30 pmol · g^-1), although this was still the predominant cytokinin. [9R]Z and [9G]Z were present at between 2 and 15 pmol · g^-1, with iP at 1–5 pmol · g^-1 and [9R]iP at between 1 and 12 pmol · g^-1. Seedlings contained similar amounts with the exception of a lower [9G]iP content (5–10 pmol · g^-1) and more [9R]iP (10–20 pmol · g^-1). Root tissues of ramets contained significantly higher concentrations of [9G]iP than shoots. Comparison of two isogenic lines of one clone giving rise to normal and abnormal palms showed significantly higher concentrations of [9R]Z and [9G]Z in the normal than in the abnormal line and, in embryoids only, higher [9G]iP in the normal line. In all other cases the between-done differences were greater than any normal/abnormal differences. There was a general tendency for increased concentrations of [9G]iP in abnormal lines and for this compound to be in a higher concentration in embryoids and plants derived from culture than in zygotic embryos and seedlings. Analysis of cytokinins in immature female inflorescences of normal and abnormal palms of a single clone showed the abnormal inflorescences to have higher concentrations of [9R]Z and [9R]DHZ and less [9G]Z than the normal inflorescences at comparable stages of development.Abbreviations HPLC high performance liquid chromatography - [9G]iP 9-glucoside of isopentenyladenine - [9G]Z 9-glucoside of zeatin - [9R]Z zeatin riboside - iP isopentenyladenine - [9R]iP isopentenyladenosine - [9G]DHZ dihydrozeation-9-glucoside - ELISA enzyme-linked radioimmunosorbentassay - ANOVAR analysis of variance     

Further iridoids from the roots of Patrinia scabra

Two new iridoids, 1,3-dimethoxy-4,7-dimethyl-octahyhro-cyclopenta[c]pyran-6,7-diol (1) and isovillosol (2), together with two known compounds, were isolated from the roots of Patrinia scabra. Their structures were elucidated by different spectroscopic methods including IR, HRESIMS, 1D and 2D NMR experiments as well as X-ray crystallography.     

Hypoglycemic activity of a novel anthocyanin-rich formulation from lowbush blueberry,Vaccinium angustifolium Aiton

Blueberry fruits are known as a rich source of anthocyanin components. In this study we demonstrate that anthocyanins from blueberry have the potency to alleviate symptoms of hyperglycemia in diabetic C57b1/6J mice. The anti-diabetic activity of different anthocyanin-related extracts was evaluated using the pharmaceutically acceptable self-microemulsifying drug delivery system: Labrasol. Treatment by gavage (500 mg/kg body wt) with a phenolic-rich extract and an anthocyanin-enriched fraction formulated with Labrasol lowered elevated blood glucose levels by 33 and 51%, respectively. The hypoglycemic activities of these formulae were comparable to that of the known anti-diabetic drug metformin (27% at 300 mg/kg). The extracts were not significantly hypoglycemic when administered without Labrasol, demonstrating its bio-enhancing effect, most likely due to increasing the bioavailability of the administered preparations. The phenolic-rich extract contained 287.0±9.7 mg/g anthocyanins, while the anthocyanin-enriched fraction contained 595±20.0 mg/g (cyanidin-3-glucoside equivalents), as measured by HPLC and pH differential analysis methods. The greater hypoglycemic activity of the anthocyanin-enriched fraction compared to the initial phenolic-rich extract suggested that the activity was due to the anthocyanin components. Treatment by gavage (300 mg/kg) with the pure anthocyanins, delphinidin-3-O-glucoside and malvidin-3-O-glucoside, formulated with Labrasol, showed that malvidin-3-O-glucoside was significantly hypoglycemic while delphinidin-3-O-glucoside was not.     

Influence of White Clover Mosaic Potexvirus Infection on the Endogenous Cytokinin Content of Bean

The cytokinin content in the primary leaves of bean (Phaseolus vulgaris) was monitored for 10 d after inoculation with white clover mosaic potexvirus. The cytokinins were isolated, purified, separated by high-performance liquid chromatography, and quantified by radioimmunoassay. The cytokinins detected at the time of inoculation (d 0) were: (a) the free bases, zeatin (Z), dihydrozeatin (DZ), and isopentenyladenine; (b) the riboside, DZ riboside (DZR); (c) the O-glucosides of DZ, DZR, and Z riboside; (d) the nucleotides, Z riboside-5′-monophosphate and isopentenyladenosine-5′-monophosphate; and (e) trace amounts of Z-9-glucoside and DZ-9-glucoside. During the 10 d after inoculation with white clover mosaic potexvirus, marked quantitative changes in this cytokinin profile were observed. The concentration of the free bases and DZR decreased, accompanied by an increase in the 9-glucosides and the nucleotides. Virus titer increased rapidly 3 d after inoculation, attaining a maximum level at d 5. This increase coincided with the increases in the 9-glucosides and the nucleotides. We propose that the decline in the cytokinin free bases and riboside may allow the increase of virus titer in bean and lead to the senescence of infected leaves.     

Flavonoid constituents of Ephedra alata

Two new flavonol glucosides have been identified in Ephedra alata, namely, herbacetin 8-methyl ether 3-O- glucoside-7-O-rutinoside and herbacetin 7-O-(6″-quinylglucoside). The known flavonoids vicenin II, lucenin III, kaempferol 3-rhamnoside, quercetin 3-rhamnoside and herbacetin 7-glucoside were also found. The structure of the isolated compounds was determined mostly by FABMS and ¹H NMR spectroscopy. The final structure of the new compounds and of herbacetin 7-glucoside was confirmed by ¹³C NMR spectroscopy.     

Isolation, structural elucidation, and neuroprotective effects of iridoids from Valeriana jatamansi

Two new iridoids, jatadoids A (1) and B (2), and two known compounds (3 and 4) were isolated from Valeriana jatamansi. Their structures were elucidated on the basis of extensive spectroscopic analyses (IR, ESI-MS, HR-ESI-MS, 1D and 2D NMR). Compound 1 possessed an isovaleroxy group at the C-3 position that has previously been unreported in the class of iridoids. Four compounds were evaluated and compounds 1 and 3 showed moderate neuroprotective effects against MPP⁺-induced neuronal cell death in human dopaminergic neuroblastoma SH-SY5Y cells.     

The distribution of fatty acids including petroselinic and tariric acids in the fruit and seed oils of the Pittosporaceae, Araliaceae, Umbelliferae, Simarubaceae and Rutaceae

The fatty acid composition of the fruit oils or seed oils of Pittosporaceae (eight genera, 10 species), Araliaceae (two species), Simarubaceae (three species), and of one umbelliferous and one rutaceous species were determined by gas chromatography, argentation TLC and ozonolysis. In the Pittosporaceae, in which the major C₁₈ fatty acid of all species was either oleic acid (18:1, 9c) or linoleic acid (18:2, 9c, 12c), large amounts of C₂₀ and C₂₂ fatty acids seem to occur regularly. Petroselinic (18:1, 6c) and tariric (18:1, 6a) acids were absent. However, petroselinic acid was the major fatty acid in the Araliaceae and Umbelliferae. In these two families only small amounts of C₂₀ and C₂₂ acids were detected and tariric acid was absent. The Rutales contained relatively high amounts of trans-octadecenoic acids (18:1, 9t). Tariric acid was the major fatty acid in the two species of Picramnia (Simarubraceae), which also contained small amounts of petroselinic acid. The major fatty acids in Ailanthus glandulosa (Simarubaceae) and Phellodendron amurense (Rutaceae) were linoleic or linolenic acid (18:3, 9c, 12c, 15c); these species contained neither tariric nor petroselinic acid and the levels of C₂₀ and C₂₂ fatty acids were low. The appearance of schizogenous resin canals and polyacetylenes and the absence of iridoids and petroselinic acid allows the Pittosporaceae to be separated from the Rutales and Araliales and to be placed in an independent order, the Pittosporales. Arguments for a rather close relationship of the Pittosporales to the Araliales and Cornales (including the Escalloniaceae) are presented.     

Polyphenols of Eucalyptus sideroxylon wood

Twenty-three major components were detected in the methanol extractives of the heartwood of Eucalyptus sideroxylon. The components identified include resveratrol, resveratrol-β-glucoside, 3,3′-di- and 3,3′,4-tri-o-methylellagic acids and their glucosides. The 3,3′-di-o-methylellagic acid 4′-glucoside isolated had properties significantly different from those previously reported for this compound. Also present were gailic acid, catechin, ellagic acid, an unidentified stilbene, the ellagitannins D-6 and D-13, polymerized leucocyanidin and an oily material. The sapwood contained gailic acid, small amounts of ellagitannins and ellagic acids and traces of other components. The heartwood extractives of related eucalypt species were also examined.     

Iridoids and alkaloids from Castilleja host plants for platyptilia pica. Rhexifoline content of P. pica

Iridoid and alkaloid analyses were conducted on Castilleja sulphurea, C. occidentalis, C. rhexifolia and C. hispida (Scrophulariaceae). Pyrrolizidine alkaloids were found in C. rhexifolia and some C. sulphurea populations, but other C. sulphurea populations had quinolizidine alkaloids or none at all. No C. occidentalis populations contained alkaloids. C. hispida was found to contain lamprolobine and the quinolizidine alkaloid anagyrine. All taxa contained the pyridinemonoterpene rhexifoline. The iridoid content of the Castilleja species were all qualitatively similar. Major iridoids were aucubin, catalpol, penstemonoside and shanzhiside methyl ester, with traces of 8-epiloganin and gardoside methyl ester. Larvae of Platyptilia pica (Pterophoridae) hosted by Castilleja were found to excrete and not sequester iridoids. The adult moths contained rhexifoline alkaloid, but at a low concentration level. Systematic implications of the results for Castilleja are discussed.     

Chemical constituents from Gentianella turkestanorum (Gentianaceae)

Phytochemical investigation of Gentianella turkestanorum (Gentianaceae) afforded nineteen compounds, including six xanthones (1–6), two triterpenoids (7–8), eight flavones (9–16) and three iridoids (17–19). Here, we firstly reported that 1-hydroxy-3,5-dimethoxyxanthone (4), 1, 8-dihydroxy-3-methoxyxanthone (5), apigenin (9), quercetin (10), luteolin-7-O-glucoside (12) and three other compounds (1, 8-dihydroxy-3-methoxyxanthone (5), apigenin-7-O-gluco (1″ → 3‴) glucoside (15) and luteolin-7-O-gluco (1″ → 3‴) glucoside (16)) could be isolated from G. turkestanorum. The occurrence of chemical data and the sequence data might be employed as common constituents of the genera Gentianella, Lomatogonium and Swertia.     

Protein subunits and amino acid composition of wild lentil

Three wild species of lentil, Lens orientalis, L. ervoides and L. nigricans were investigated for protein subunits of the albumin protein fraction (APF), globulin protein fraction (GPF) and for protein and free amino acid composition. The APF and GPF formed 12.7–16.8 % and 34.7–49.0 %, respectively, of the meal nitrogen. SDS-PAGE showed APF to contain 15 to 20 major and a similar number of minor protein subunits ranging in M_r at least from 14 400 to 94 000. The GPF was also heterogenous and contained some subunits having M_r similar to APF subunits but none < 15 000. The three wild lentil species were distinguishable by their protein subunit composition. The protein amino acid composition of the wild species was identical and similar to that of the cultivated lentil. The wild species, like the cultivated species (L. culinaris), contained major amounts of free arginine, glutamic and aspartic acids, serine and a number of unidentified amino acids. L. orientalis, L. nigricans and the cultivated lentil contained two acidic and two basic unidentified amino acids. However, L. ervoides was distinctly different in that it contained only the two acidic plus one neutral unidentified amino acid, but none of the two basic unidentified amino acids.     

Blueing of sepal colour of Hydrangea macrophylla

Blue and red sepals of Hydrangea macrophylla were quantitatively analyzed for aluminium, anthocyanin (delphinidin 3-glucoside) and copigments (caffeoyl- and p-coumaroylquinic acids). All the blue sepals examined contained both Al and copigments (especially 3-caffeoylquinic acid) in considerable amounts. In in vitro experiments using 3- and 5-caffeoylquinic acids, Al and delphinidin 3-glucoside, it was shown that 3-caffeoylquinic acid and Al formed a blue complex with the anthocyanin. Absorption spectra of the blue complex were practically identical with those of the blue solutions obtained from blue hydrangea sepals by extraction with 4 M NACl. In contrast, 5-caffeoylquinic acid (chlorogenic acid) which was also present in hydrangea sepals gave only a red-purple colour with Al and the anthocyanin. Neither 3-caffeoylquinic acid nor Al independently produced blue colour when mixed with the anthocyanin in the mole ratios of 1–30, this being the range that the compounds were found in blue sepals. These results suggest that blue colour of hydrangea sepals is due mainly to the blue complex of delphinidin 3-glucoside-aluminium-3-caffeoylquinic acid. The role of aluminium may be to stabilize an interaction between the quinic ester and the anthocyanin.