Identification and characterization of a rhamnosyltransferase involved in rutin biosynthesis in Fagopyrum esculentum (common buckwheat)

Rutin, a 3-rutinosyl quercetin, is a representative flavonoid distributed in many plant species, and is highlighted for its therapeutic potential. In this study, we purified uridine diphosphate-rhamnose: quercetin 3-O-glucoside 6″-O-rhamnosyltransferase and isolated the corresponding cDNA (FeF3G6″RhaT) from seedlings of common buckwheat (Fagopyrum esculentum). The recombinant FeF3G6″RhaT enzyme expressed in Escherichia coli exhibited 6″-O-rhamnosylation activity against flavonol 3-O-glucoside and flavonol 3-O-galactoside as substrates, but showed only faint activity against flavonoid 7-O-glucosides. Tobacco cells expressing FeF3G6″RhaT converted the administered quercetin into rutin, suggesting that FeF3G6″RhaT can function as a rhamnosyltransferase in planta. Quantitative PCR analysis on several organs of common buckwheat revealed that accumulation of FeF3G6″RhaT began during the early developmental stages of rutin-accumulating organs, such as flowers, leaves, and cotyledons. These results suggest that FeF3G6″RhaT is involved in rutin biosynthesis in common buckwheat.     

Stereochemistry of C-methylation in the biosynthesis of rhododendrin in Alnus and Betula

Using differently labelled precursors, it was established that rhododendrin (3-(4-hydroxyphenyl)-1-methylpropyl-β-D-glucopyranoside) is formed through the phenylpropane pathway via p-coumaryl alcohol, dihydro-p-coumaryl alcohol and C-methylation of the γ-C-atom of the C₆C₃ unit with methionine supplying the methyl group. It was demonstrated that the pro-(S)-hydrogen atom of dihydro-p-coumaryl alcohol is replaced stereospecifically by the methyl group.     

The biosynthesis of melanin in Alternaria

A melanin which is insoluble in strong alkali has been isolated from Alternaria mycelium. Alkali fusion of the pigment produced p-hydroxybenzoa     

The biosynthesis of abscisic acid in Cercospora rosicola

1′-Deoxyabscisic acid (1′-deoxy-ABA) has been isolated from cultures of Cercospora rosicola which are actively synthesizing abscisic acid (ABA)     

The biosynthesis of angelic acid in Cynoglossum officinale

Six-month-old Cynoglossum officinale plants were fed via the roots with tiglic acid-[1-¹⁴C]. After a week the plants were harvested, the alkaloid heliosupine was isolated and hydrolysed to heliotridine and angelic acid. The latter contained all the radioactivity of the original heliosupine showing that angelic acid may be formed from tiglic acid by a cis-trans isomerization.     

Kaurene and kaurenol biosynthesis in cell-free system of Phaseolus coccineus suspensor

It is shown that suspensor tissue of Phaseolus coccineus can biosynthesize ent-kaur-16-ene and ent-kaur-16-en-19β-ol, two key precursors in the biosynthesis of gibberellins.     

Pharmacokinetics of chlorogenic acid and rutin in larvae of Heliothis zea

Studies using [³H]chlorogenic acid and [³H]rutin demonstrated that the kinetics of uptake of these plant phenolics into the haemolymph of 5th-instar Heliothis zea (Boddie) following actue oral administration is a first-order process. The total quantity of either phenolic present in the haemolymph within 1 hr amounts to 5% or less of the total ingested dose. Based on TLC analyses, 80% or more of the radioactivity in the haemolymph occurs as the parent phenolic. Retention of [³H]-chlorogenic acid or [³H]-rutin in H. zea following chronic feeding from 1st to 3rd-instar larvae is also linearly related to dietary dose. Chlorogenic acid and rutin are both equitoxic and equivalent in bioavailability to H. zea.Loss of [³H]-rutin from the haemolymph of 5th-instar larvae following injection is biphasic. One half of the injected dose is excreted in the frass in the first 6 hr after injection; the other half is thereafter eliminated at 1/20th of the initial rate. Analyses of extracts of frass by thin-layer chromatography indicate that after either chronic or acute feeding 90% of the ingested phenolic is excreted unchanged. Possible sites and modes of action of phenolics in insects are discussed in light of these findings.     

Differences in the incorporation pattern of labelled cinnamic acid and l-phenylalanine into the anthocyanins of Petunia hybrida

The incorporation of phenylalanine-[¹⁴C] into anthocyanins of petals of Petunia hybrida is greater than that of cinnamic acid-[¹⁴C]. Moreover, there is a preferential incorporation of phenylalanine-[¹⁴C] into delphinidin 3-monoglucoside, as compared with the incorporation into cyanidin and peonidin 3-monoglucosides.     

Structure and biosynthesis of 5-deoxyfusarubin and anhydro-5-deoxyfusarubin,naphthaquinone pigments from Nectria haematococca

Two naphthaquinone pigments, 5-deoxyfusarubin and anhydro-5-deoxyfusarubin, were isolated from a yellow strain of the fungus Nectria haematococca. The ¹³C NMR spectra of 2 biosynthetically labelled from [1-¹³C]acetate confirmed the structures of the pigments and established the heptaketide origin of the molecules.     

In vitro and in vivo biosynthesis of xanthophylls by the cyanobacterium Aphanocapsa

Of the six carotenoids identified in the cyanobacterium Aphanocapsa, β-carotene, zeaxanthin, echinenone and myxoxanthophyll are the major pigments, whilst β-cryptoxanthin and 3-hydroxy-4-keto-β-carotene are present only in trace amounts. With the exception of zeaxanthin, the other xanthophylls could be formed in vitro from [¹⁴C]phytoene in high yields, especially β-cryptoxanthin and 3-hydroxy-4-keto-β-carotene. In a time course experiment of xanthopyll biosynthesis the flow of radioactivity from [¹⁴C]phytoene was followed through the pools of phytofluene, lycopene, and β-carotene. The reaction sequence from phytoene to xanthophylls is sensitive in vitro to both difunone, an inhibitor of carotene desaturation, and CPTA, an inhibitor of cyclization.     

On the intracellular site of biosynthesis of α-tocopherol in Hordeum vulgare

When barley seedlings were exposed to ¹⁴CO₂ for 1–3 hr, ¹⁴C was preferentially incorporated into plastoquinone, but only in low amounts i     

Biosynthesis and distribution of tomatine in cultured excised Lycopersicon esculentum roots

Growth of cultured excised tomato roots in the presence of ¹⁴C-mevalonic acid lactone results in labelling of tomatine. In the main axis of the root, incorporation was greatest in the apical meristematic region. Tomatine was present in equal concentrations in all parts of the cultured root system.     

In vitro biosynthesis of isopentenylacetophenones in Eupatorium rugosum

A cell free homogenate of Eupatorium rugosum leaves was prepared and utilized to study the biosynthesis of dehydrotremetone (1). Homogenates of young leaves were most efficient in the formation of 1. The furan ring and its side chain were derived from an isoprenoid unit, which appeared to be isopentenyl pyrophosphate and not dimethylallyl pyrophosphate. Potential aromatic precursors such as 4-hydroxyacetophenone and 4-hydroxy-3[isopenten-(2)-yl]-acetophenone were poorly utilized. However, tremetone, especially in the presence of the coenzyme NADP, was very efficiently converted to 1. An apparent intermediate in the pathway leading to 1 was isolated and appeared similar to 4-hydroxy-3[isopenten-(2)-yl]-acetophenone. It is proposed that the aromatic moiety of 1 is derived from acetate via a polyketide intermediate, which undergoes isoprenylation by isopentenyl pyrophosphate, followed by aromatization and furan ring closure.     

Specificity effects in the biosynthesis of fatty acids in Bacillus acidocaldarius

Addition to Bacillus acidocaldarius of acids which can act as primers for fatty acid synthesis promote the synthesis of corresponding fatty acids competitively. The effective acids are n?C₅ to -?₇ (not C₄ or C₈), iso- and anteiso-C, and ?C, (not C4), and a range of cyclic acids from cyclobutylacetic and cyclopentanecarboxylic to cycloheptylacetic. New non-natural ω-cyclobutyl-, ω-cyclopentyl-, and ω-cycloheptyl-fatty acids are obtainable. The range of acceptable primers and the range of fatty acids produced therefrom indicate, respectively, the substrate specificities of the transacylase which introduces acyl species into fatty acids synthesis and the one which removes them. The specificity of the primer transacylase may be similar to that in some rumen anaerobes.     

Hormonal control of sterol biosynthesis in Phaseolus aureus

Naphthalene acetic acid increased the sterol content of mung bean hypocotyl sections mainly in the zone of elongation growth. The increased sterol synthesis can be explained by a stimulated conversion rate of cycloartenol into sterols. During the 20-hr incubation period the stigmasterol: sitosterol ratio increased considerably.     

The biosynthesis of long-chain hydrocarbons in the green alga Botryococcus braunii

The green colonial alga Botryococcus braunii has unusually high levels of hydrocarbons. Two distinct sites of hydrocarbon accumulation are present in the species: an internal pool present in cytoplasmic inclusions and an external pool in the trilaminar outer walls and associated globules. It is generally assumed that the hydrocarbons are produced within the cells and then excreted into the external pool to maintain the intracellular content at a normal value. Various feeding experiments showed, however, that the radioactivity of the external pool is much higher than the internal one. On the other hand, there was no decrease in the labelling of internal hydrocarbons in chase experiments. Therefore, an excretory process apparently does not take place in B. braunii. The outer wall, therefore, is the main site of hydrocarbon accumulation and also the place where the bulk of B. braunii hydrocarbons are produced. The outer wall also is involved in the matrix of colony formation and the above findings account for the sharp decrease of hydrocarbon production which is associated with the loss of colonial habit. The cultures were also shown to be unable, under usual growth conditions, to catabolize their own hydrocarbons. Such a feature, along with the extracellular location of the main site of production, may account for the abnormally high content of hydrocarbons typical of B. braunii.     

Kinetic study of possible intermediates in the biosynthesis of chlorogenic acid in Cestrum poeppigii

p-Coumaric and 3-O-p-coumarylquinic acid seem to be important precursors of chlorogenic acid in the leaves of Cestrum poeppigii. 3-O-Cinnamylquinic acid, which has a very small metabolic activity, is of little importance in this respect. The kinetics of incorporation of radioactivity from t-cinnamic acid-3-[¹⁴C] into p-coumaric, 3-O-p-coumarylquinic, chlorogenic and 3-O-cinnamylquinic acid showed that the biosynthetic rates for these products decrease in the order shown. For p-coumaric acid, which has a markedly high metabolic activity, a turnover rate of 28 μg/hr and per gram fresh plant leaf, was calculated. Some trapping experiments with caffeic acid, and the acids mentioned above and using either t-cinnamic acid-3-[¹⁴C] or p-coumaric acid-2-[¹⁴C] as precursor, are discussed. A HPLC method for the rapid determination of phenolic acids in plant extracts, is described.     

Biosynthesis of furanosesquiterpenoid stress metabolites in Ipomoea batatas: Isotopic oxygen incorporation into ipomeamarone

All three oxygen atoms in ipomeamarone, a major stress metabolise of the sweet potato, were found to originate from molecular oxygen and not from water or the sesquiterpene precursor, farnesol.     

The biosynthesis of feruloyltyramine in Nicotiana tabacum

The biosynthesis of feruloyltyramine in Nicotiana tabacum Xanthi n.c. leaves is achieved through the action of the enzyme feruloyl-CoA tyramine N-feruloyl-CoA transferase. Its activity is increased 5- to 8-fold following infection by tobacco mosaic virus at 20°. The enzyme is soluble, its MW is 45 000, and it can synthesize a wide range of amides due to its low specificity for cinnamoyl-CoA thioesters and aromatic amines. Its affinity for feruloyl-CoA fragments is also described.     

Isoflavone biosynthesis in Onobrychis viciifolia: Formononetin and texasin as precursors of afrormosin

4,2′,4′-Trihydroxychalcone- [carbonyl-¹⁴C], formononetin- [Me-¹⁴C] and texasin- [Me-¹⁴C] were all good precursors of afrormosin (7-hydroxy-6,4′-dimethoxyisoflavone) in Onobrychis viciifolia seedlings, and a biosynthetic pathway involving these intermediates is proposed. 2′,4′-Dihydroxy-4-methoxychalcone- [carbonyl-¹⁴C] and daidzein-[carbonyl-¹⁴C] were poor precursors. Incorporations into formononetin were also recorded.