Lectin variability in Phaseolus coccineus

Lectin variability within Phaseolus coccineus is revealed by non-denatured electrophoretic patterns and immunological labelling of total seed protein extracts, showing that the different cultivars and wild varieties studied can be classified into three main categories according to the number of isolectins (three, two or one) present in each extract. Attempts in the purification of these isolectins were performed on three different affinity systems in which ligands were thyroglobulin (known to purify the P. vulgaris isolectins), pig red cell membrane ghosts (stroma) or antibodies against the P. vulgaris cv. Contender E₂L₂ isolectin. The P. coccineus isolectins exhibit varied affinities towards thyroglobulin and stroma, the cathodic and anodic (pH 4.5) isolectins being respectively retained by the two systems, whereas the antibody affinity system is the only one able to purify the totality of the isolectins present in an extract.     

Isoflavonoids from Phaseolus coccineus

After treatment with CuCl₂, the following isoflavonoids have been isolated from the runner bean, Phaseolus coccineus: daidzein, genistein, isoprunetin, 2′-hydroxygenistein, phaseoluteone, 2′-hydroxydihydrodaidzein, isoferreirin, kievitone, cyclokievitone, glycinol, phaseollidin, phaseollin, demethylvestitol, phaseollinisoflavan, 2′-hydroxyisoprunetin and 7,4′-dihydroxy-5,2′-dimethoxyisoflavanone. The latter two compounds are novel natural products.     

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.     

Squalene biosynthesis by a cell-free extract of Rhizopus arrhizus

Squalene was identified as the principal radioactive component (85%) of the hexane-soluble products formed from mevalonate-[2-¹⁴C] by a cell-free preparation from Rhizopus arrhizus. The system required ATP, NADH and Mn²⁺ to obtain 20–40% incorporation of the substrate into squalene. Temperature and pH optima for the system were 20° and 7.0, respectively.     

Characterization of gibberellins from light-grown Phaseolus coccineus seedlings by combined GC-MS

Following extensive purification of an extract from 5000 light-grown Phaseolus coccineus seedlings by procedures including countercurrent distribution, Sephadex G10, polyvinylpyrrolidone, charcoal-celite and silicic acid partition column chromatography, TLC preparative GLC, gibberellins A₁, A₄, A₅ and A₂₀ were characterized by combined GC-MS. In addition, an unknown compound isomeric with gibberellin A₁₇ but lacking an hydroxyl group, was also detected.     

Polyoxygenated ent-kauranes and water-soluble conjugates in seed of Phaseolus coccineus

C-α and C-β, previously isolated from seed of Phaseolus coccineus, are shown respectively to be the bis-O-isopropylidene and the 16,17-mono-O-isopropylidene derivatives of ent-6α,7α,16β,17-tetrahydroxykauranoic acid. By GC-MS characterization of the products of acidic, basic and enzymatic hydrolysis, water soluble conjugates of the following compounds have been shown to occur in P. coccineus seed: GA₈, GA₁₇, GA₂₀, GA₂₈, ent-6α,7α,13-trihydroxykaurenoic acid, ent-6α,7α,17-trihydroxy-16β-kauranoic acid, ent-6α,7α,16β,17-tetrahydroxykauranoic acid, 7β,13-dihydroxykaurenolide and abscisic acid.     

Cell wall glycoproteins and polysaccharides of parenchyma of Phaseolus coccineus

Fractionation of the cell wall material of parenchyma of mature runner beans with and without chlorite-HOAc treatment, clearly showed that at least two main types of wall proteins were present. One relatively rich in hydroxyproline (HP) associated with α′-cellulose, from which most (90%) of it could be readily liberated by chlorite-HOAc treatment and the other relatively poor in HP associated with hemicellulose A. The chlorite HOAC solubilized “glycoprotein” contained a high proportion of arabinose and galactose. It was purified by PhOH-H₂O fractionation and the molar ratios of HP, arabinose, galactose, xylose, rhamnose, glucose and uronic acid in the purified glycoprotein (“glycoprotein X”) were 1:2·6:2·4:0·2:0·2:0·1:0·3. The principal amino acids of glycoprotein X were HP (43·5 mol%), serine and proline which together comprised 66 mol% of the total. These results suggest that the HP-rich wall glycoprotein is associated with cellulose microfibrils and approximates in conformation to polyhydroxyproline carrying arabinose and galactose oligosaccharide side chains.     

Biosynthesis of ent-kaurene in cell-free extracts of Pisum sativum shoot tips

Soluble enzyme preparations from pea shoot tips incorporated mevalonic acid-2-¹⁴C into ent-kaurene-¹⁴C, squalene-¹⁴C and other products. The assay for either ent-kaurene or squalene is quite direct; both products can be obtained apparently free of radioactive contaminants by TLC on silica gel G in hexane. The enzyme system is dependent upon added ATP and Mn²⁺ or Mg²⁺, with Mn²⁺ being a more effective activator than Mg²⁺ under the experimental conditions. Reduced pyridine nucleotide had no effect on ent-kaurene production but stimulated squalene synthesis. The accumulation of both ent-kaurene and squalene was stimulated by dithiothreitol and carbon monoxide and was reduced by the addition of particulate cell components. AMO-1618 inhibited ent-kaurene production and had no effect on the synthesis of squalene. Enzyme extracts from shoot tips are much less active in ent-kaurene synthesis than extracts from the cotyledons of immature seeds on either a fresh weight or protein basis.     

Biosynthesis of the phytoalexin phaseollin in Phaseolus vulgaris

Feeding experiments in CuCl₂-treated French bean (Phaseolus vulgaris) seedlings have demonstrated that labelled 2′,4′,4-trihydroxychalcone, daidzein, 7,2′,4′-trihydroxyisoflavone, 3,9-dihydroxypterocarpan and phaseollidin are all good precursors of the pterocarpan phytoalexin phaseollin. These compounds represent a logical sequence in the biosynthetic pathway to phaseollin.     

Isolation of legumin-like protein from Phaseolus aureus and Phaseolus vulgaris

An 11S seed globulin has been isolated from Phaseolus aureus and P. vulgaris by zonal isoelectric precipitation and the MWs of the constituent subunits determined. The protein of P. vulgaris occurs in the protein body fraction and its chemical composition, including the N-terminal amino acids and amino acid composition has been determined. The similarity between the 11S globulin of the two Phaseolus spp. and legumin from other leguines is discussed.     

Cyanoglucoside biosynthesis in white clover (Trifolium repens)

l-valine-(U-¹⁴C), isobutyraldoxime-(U-¹⁴C) and 2-hydroxyisobutyraldoxime-(1,3-¹⁴C) were fed to white clover shoots which contained or lacked cyanoglucoside. Labelling results indicated that plants which were unable to synthesize linamarin from these precursors lacked the ability to glucosylate 2-hydroxyisobutyronitrile. When l-valine-(U-¹⁴C) was fed to cyanoglucoside-producing plants in the presence of cold isobutyraldoxime, radioactivity could be trapped in this compound. No radioactivity was trapped in isobutyraldoxime fed to non-cyanoglucoside-producing shoots demonstrating that this phenotype also lacks the ability to convert valine into the aldoxime. This suggests that the non-cyanoglucoside-producing plants have at least two steps in the biosynthesis of linamarin missing. Differences between the linamarin biosynthetic enzymes of white clover and flax were demonstrated using the inhibitor O-methylthreonine and by the failure of cyanoglucoside-producing white clover shoots to synthesize linamarin from fed 2-hydroxyisobutyraldoxime.     

The effect of exogenous gibberellic acid on gibberellin biosynthesis by Gibberella fujikuroi

The addition of gibberellic acid and some other gibberellins to cultures of Gibberella fujikuroi suppresses the incorporation of [2-¹⁴C]MVA and ¹⁴C-labelled ent-kaurene into the gibberellin metabolises.     

Biosynthesis of coumestrol in Phaseolus aureus

Feeding experiments with 4′,7-dihydroxyisoflavone-[4-¹⁴C] (daidzein), 2′,4′,7-trihydroxyisoflavone-[T] and (±)-4′,7-dihydroxyisoflavanone-[T] (dihydrodaidzein) in suspension cultures of mung bean (Phaseolus aureus Roxb.) roots have shown that daidzein is a better precursor of the coumestan coumestrol than is the trihydroxyisoflavone and that dihydrodaidzein can also be converted very efficiently. The results provide further evidence for the intermediacy of a pterocarp-6a-en in coumestrol biosynthesis, and also indicate the possible existence of a 'metabolic grid' of isoflavones and isoflavanones in P. aureus.     

The fate of C-20 in C19 gibberellin biosynthesis

Experiments with ent-kaur-16-ene-[¹⁴C], prepared biosynthetically from sodium acetate-[2-¹⁴C], have shown that the C-20 carbon atom of the C₂₀ gibberellins is evolved as carbon dioxide during the biosynthesis of the C₁₉ gibberellins by Gibberella fujikuroi.     

The stereospecificity of biosynthesis of squalene and β-amyrin in Pisum sativum

The distribution of deuterium in squalene and β-amyrin, biosynthesized from mevalonic acid-6,6,6-d₃ in Pisum sativum, has been examined b     

Protein-polysaccharide linkages in glycoproteins from Phaseolus vulgaris

Serine and hydroxyproline participate in protein-polysaccharide linkages in hydroxyproline-poor glycoproteins from Phaseolus vulgaris cv Pinto. Most substituted hydroxyproline residues contain arabinose, galactose and glucose, but some have arabinose only. Serine residues contain arabinose, galactose and glucose.     

Attempts to demonstrate incorporation of labelled precursors into acetylcholine by Phaseolus vulgaris seedlings

The labelling of acetylcholine was investigated in bean seedlings (Phaseolus vulgaris) and hypocotyl hooks. Labelling could only be found after red light irradiation but not in the dark. Acetate was a much better precursor than choline. Glucose gave no detectable amounts of acetylcholine labelling during 24 hr of incubation. The rate of acetylcholine labelling was higher in whole seedlings than in hypocotyl hook preparations.     

Metabolic transformations of some ent-kaurenes in Gibberella fujikuroi

The conversion of ent-kaur-16-enes to gibberellic acid in Gibberella fujikuroi is blocked by A-ring modifications. Thus ent-3β-hydroxykaur-16-en-19-yl succinate gives good conversion (46%) to the 7β-hydroxy derivative.* Under the same conditions the 3β-epimer gives 7β- or 6α-hydroxylation and the former occurs for the 3-oxo analogue. The succinoyloxy function acts as a less efficient block and ent-kaur-16-en-19-yl succinate is converted to 7β-hydroxy and 6β,7β-dihydroxy derivatives along with gibberellic acid. Hydrolysis of the succinate block of the metabolities provides the 7β, 19-diol and 6β,7β, 19-triol. Of this pair only the former was effectively metabolized to gibberellic acid in G. fujikuroi.     

Starch synthesis in isolated Phaseolus vulgaris chloroplasts

Isolated bean (Phaseolus vulgaris) chloroplasts were used to investigate the mode of synthesis of transitory amylose and amylopectin from ADP-glucose. Pulse chase experiments showed that labelled glucose in amylose decreased when chased with cold substrate as compared to controls. A significant portion of this decrease appeared in the amylopectin fraction indicating that amylopectin was formed from amylose. However, time course experiments showed that the rate of amytopectin synthesis is higher than that of amylose at the early stages of incubation, suggesting a certain degree of independent synthesis of the two fractions. High concentration of citrate increased the rate of amylopectin synthesis.     

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.