Effect of diethylpyrocarbonate and sodium deoxycholate on polyribosomes from cotyledons of Vicia faba

Degradation of rRNA during the isolation of polyribosomes in the presence of either sodium deoxycholate or diethylpyrocarbonate gave breakdown products with MW corresponding to 1·13, 1·02, 0·93, 0·79, 0·61, 0·52, 0·44, 0·42, 0·30, 0·28, 0·24, 0·20 and 0·12 x 10⁶ daltons.     

A comparison of acid extracted globulin fractions and vicilin and legumin of Vicia faba

Two globulin fractions, obtained by extraction using acidic conditions, were characterized by their sedimentation properties, subunit compositions and     

Multiple forms of Vicia faba α-galactosidases and their relationships

Three highly purified α-galactosidases, I, II¹ and II² have been isolated from resting Vicia faba seeds. Form I (MW 160 000) is a tetrame     

Dihydrowyerone derivatives as components of the furanoacetylenic phytoalexin response of tissues of Vicia faba

Changes in concentrations of 7 wyerone derivatives in bean tissues undergoing resistant reactions to Botrytis cinerea or B. fabae and in cotyledons in response to mercuric chloride have been examined using high performance liquid chromatography. The proportion of derivatives occurring in their saturated (dihydro) forms varied between cotyledon, leaf and pod tissues and with time after inoculation. Unsaturated derivatives were always present in greater concentrations than their dihydro analogues.     

Specific subunit pairs of legumin from Vicia faba

Four pairs of disulphide-linked acidic (α) and basic (β) subunits were isolated from legumin of Vicia faba. Pairing between α- and β-subunits is nonrandom, supporting the view that each subunit pair arises from a common precursor polypeptide, already containing intramolecular disulphide bonds, when cleavage to the subunit pair takes place. The subunit pairs belong to two structural types: type A contains Met, whereas type B lacks Met. In addition to these four subunit pairs, at least two more pairs are present in legumin in minor amounts.     

Biosynthesis of 3,4-dihydroxyphenylalanine in Vicia faba

Radioactive shikimic acid and l-tyrosine were shown to be efficient precursors of 3,4-dihydroxyphenylalanine (DOPA) in Vicia faba. [1-¹⁴C]Acetate and l[U-¹⁴C]phenylalanine were not incorporated into tyrosine or DOPA. Thus the synthesis of DOPA occurs via the shikimic acid pathway and tyrosine or a very closely related metabolise. Phenolase was present in etiolated plants in much larger quantities after a brief light exposure whereas DOPA concentration was relatively constant during all stages of plant growth. Partially purified phenolase did not catalyze the conversion of tyrosine to DOPA and does not appear to have a role in DOPA synthesis.     

The effect of light intensity on galactolipid synthesis in Vicia faba leaves

The effect of light intensity upon galactolipid synthesis in Vicia faba leaf tissue was studied at two CO₂ concentrations, 0.03 and 1%. The rates of galactolipid synthesis were estimated by determining the amount of radioactivity in each of the two galactoses of digalactosyl diacylglycerol (DGDG) and the single galactose of monogalactosyl diacylglycerol (MGDG), a technique based upon the accepted pathway for galactolipid synthesis in which galactosylation is the terminal step in biosynthesis. The results suggest that the rates of MGDG and DGDG synthesis were similar under all conditions and that galactolipid synthesis was not directly affected by light intensity. The quantity of radioactivity incorporated into the galactoses of individual molecular species of MGDG and DGDG were similar under the light conditions used.     

Ornithine carbamyltransferase activity from the cotyledons of developing and germinating seeds of Vicia faba

During maturation the ornithine carbamyltransferase activity from cotyledons of Vicia faba sharply decreased. It declined further during subsequent germination. On the other hand, arginase activity was low in mature, air-dry seeds but increased considerably during germination. After centrifugation at 40 000 g, more than 90% of the ornithine carbamyltransferase activity remained in the supernatant. The fractions containing tightly coupled mitochondria, showed hardly any omithine carbamyltransferase activity.     

Nitrogen regime and isoenzyme changes in Vicia faba

Plants of Vicia faba were supplied either with fertilizer inorganic nitrogen or they received only nitrogen fixed by their root nodules. The esterase and transaminase isoenzyme profiles and the total water-soluble protein complement of seed cotyledons and of pollen did not vary irrespective of the N regime employed, whereas those of the leaf did. Possible causes and the implications of these differences are discussed.     

UDP-galactose 4′-epimerase from vicia faba seeds

UDP-Galactose 4′-epimerase was purified ca 800-fold through a multi-step procedure which included affinity chromatography using NAD⁺ -Agarose. Three forms of the enzyme were separated by gel-filtration but only the major form was purified. The pH optimum of the enzyme was 9.5. Exogenous NAD⁺ was not required for enzymic activity but its removal caused inactivation. The enzyme was unstable below pH 7.0 but stable at pH 8.0 in the presence of glycerol and at ?20° for two months. The equilibrium constant for the enzyme-catalysed reaction was 3.2 ± 0.15. The K_m for UDP-galactose and UDP-glucose were 0.12 mM and 0.25 mM, respectively. The inhibition by NADH was competitive, with a K_i of 5 μM. The MW of the enzyme was 78 000; the two minor forms showed the values of 158 000 and 39 000, respectively.     

Comparison of globulin proteins from Phaseolus vulgaris with those from Vicia faba

Extraction of maturing Phaseolus vulgaris seeds with an ascorbic acid—NaCI medium facilitated the preparation of two globulin fractions which wer     

Binding of α-galactosidase I from vicia faba to potato starch granules and sheep erythrocytes

α-Galactosidase I from Vicia faba seeds binds to potato starch and sheep erythrocytes. With the aid of fluorescence microscopy and using 4-methylumbelliferyl α-D-galactoside as the substrate it has been demonstrated that the binding is via the lectin sites of the enzyme leaving catalytic sites free and detectable. The lectin site is specific for D-glucose/D-mannose residues.     

Light control of periodicity of bleeding from roots of Vicia faba

The light control of the periodicity of bleeding sap was investigated, and it appears that the metabolic activity of the roots controls the exudation. Nitrite, nitrate, fructose, glucose and sucrose, various amino acids and some other unidentified compounds, were detected in the bleeding sap.     

Composition of cell walls from cotyledons of Pisum sativum,Vicia faba and Glycine max

Cell walls from cotyledons of smooth field pea, broad bean and soya bean contain ca 55% pectic polysaccharides associated with 9% cellulose. Arabinose is the major pectic sugar of pea and broad bean walls whereas soya bean pectic polymers are constituted of galactose and arabinose in the ratio (2:1). Galacturonic acid represents ca 20% of the walls. In addition, pea and broad bean cell walls contain, respectively, 12% and 6% of non-starchy and non-cellulosic glucans bearing 4,6-linked and 3-linked glycosyl units. EDTA-soluble acidic pectic substances are distinct rhamnogalacturonans bearing decreasing proportions of interrupting rhamnose from highly interrupted moieties to nearly homogenous homogalacturonans. Pea and broad bean rhamnogalacturonans are associated with arabinose-containing polymers of average DP ca 30–35 whereas soya bean ones have side chains of arabinose and galactose of DP ca 40.     

Interconversion of gibberellin A5 to gibberellin 3 in seedlings of dwarf Pisum sativum

[³H]-Gibberellin A₅ ([³H]-GA₅) applied to seedlings of dark-grown dwarf pea (Pisum sativum L. cv. Meteor), was converted to two acidic compounds, GA₃ and a chromatographically similar unknown. Identification of GA₃ was made by gas-liquid radiochromatography using three stationary phases.     

Lipoxygenase in Vicia faba minor

Lipoxygenase activity was demonstrated in partially purified preparations from small faba beans. The enzyme was shown to possess a pH optimum of 6·5 and was inactivated by exposure to 70° for 15 min. The K_m value for linoleic acid was calculated to be 0·57 mM. Ammonium sulphate fractionation yielded two highly active preparations, which were both active towards linoleic and linolenic acids. Neither fraction was inhibited by either cyanide or p-chloromercuribenzoate. The two fractions showed markedly differing responses to calcium ions, suggesting the presence of two lipoxygenases in faba beans. Activation of the enzyme by calcium ions was eliminated by the addition of EDTA.     

Fractionation and characterization of polyadenylated RNA from broad bean meristematic root cells

Several populations of polyadenylated RNA from Vicia faba méristematic root cells were fractionated by stepwise thermal elution from poly(U)-Sepharose following sequential phenol extraction. Analysis of these fractions showed that the size of the poly(A) segment could influence this fractionation, but in some cases other characteristics of the molecule are involved. Evidence was obtained that 45–60% of the nucleotides of plant polyadenylated RNA are in base paired regions, as was previously demonstrated for mammalian mRNA.

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Resume Après extraction séquentielle au phénol des ARN totaux de cellules méristématiques de racine de Fêve, les ARN polyadénylés, fixés sur colonne de poly(U)-Sépharose, sont séparés en plusieurs populations par une élution thermique à 40° et 50° C. L'analyse structurale de ces fractions montre que si la taille du segment des poly(A) joue le principal rôle dans ce partage, d'autres caractéristiques de molécules d'ARN peuvent interférer. Nous montrons de plus pour la première fois que 45 à 60% des nucléotides des ARN polyadénylés végétaux sont engagés dans des structures à double brin, comme c'est le cas dans les ARN polyadénylés animaux.

     

Formation of vicine and convicine by Vicia faba

Methods are described for the quantitative extraction and separation of the pyrimidine glucosides, vicine and convicine. The contents of these two substances in germinating seeds and young seedlings of Vicia faba remain constant for the first 2 weeks. Net synthesis and accumulation of vicine and convicine occurs in developing seeds. That the synthesis occurs within the pod and the pyrimidine glucosides are not translocated into them, was shown by injection of ¹⁴C-labelled precursors into the pods. [1-¹⁴C]- and [2-¹⁴C]-acetate were weakly incorporated but much greater incorporation was observed with [U-¹⁴C]-aspartic acid and [6-¹⁴C]-orotic acid. The results indicate that the orotic acid pathway is involved in the formation of the pyrimidine ring of both vicine and convicine.     

Protection of polyribosomes by attachment to membranes and by diethylpyrocarbonate during their extraction from Vicia faba

It was shown that membrane-bound polyribosomes of developing cotyledons wore protected during extraction by their membrane attachment and by the presen     

Interconversion of gibberellin A20 to gibberellin A29 by etiolated seedlings and germinating seeds of dwarf Pisum sativum

Tritium labelled gibberellin A₂₀ ([³H]-GA₂₀) applied to etiolated shoots and germinating seeds of dwarf pea (Pisum sativum L. cv. Meteor) was converted to gibberellin A₂₉. Identifications were made by GLRC and GC-MS.