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PubMed1984

Intervening sequences in chloroplast genomes.

Chloroplast DNAs from the green algae Euglena gracilis and from broad beans (Vicia faba) were hybridized with homologous chloroplast RNA. The hybrids were analyzed by electron microscopy. Almost all transcribed regions of the Euglena genome, except for the rRNA genes, contain intervening sequences. A minimum of 50 introns was observed amounting to about 32 kb of the 145 kb genome. Only four spliced transcripts with a total number of six introns (about 0.8 kb each) were detected in the bean chloroplast RNA. A complete transcription map of the Euglena chloroplast genome was established. Transcribed regions occur on the strand that codes for the rRNAs (plus strand) as well as on the complementary strand (minus strand). The polarity of transcription reverses at least six times.

Base Sequence↗
PubMed1975

Metabolism of trans-3-hexadecenoic acid in broad bean.

1. Broad bean (Vicia faba) leaves contain rather high concentrations (about 4% of total fatty acids) of the trans-3-hexadecenoic acid. 2. Amounts of the acid increase with the age of the leaves and are absent from etiolated tissue. 3. Changes in the levels of trans-delta-4-hexadecenoic acid can be produced by subjecting the intact plants to various light/dark periods. 4. Chloroplasts isolated from broad-bean leaves show high rates of fatty acid synthesis from [1-14C]acetate. Synthesis is dependent on coenzyme A and ATP but is insensitive to the addition of exogenous acyl carrier protein. 5. The pattern of acids made includes about 20% palmitic, 5% hexadeconoic, 10% stearic and 60% oleic. trans-3-Hexadecenoic acid synthesis was most active in chloroplasts from plants exposed to the dark for 5 days and light for 3 days. 6. Arsenite addition inhibited stearate formation by isolated chloroplasts but resulted in a two-fold stimulation of overall synthesis. 7. The rate of fatty acid synthesis by isolated chloroplasts paralleled the changes in endogenous trans-3-hexadecenoic acid levels in the leaves from which they were isolated.

Acetates↗
PubMed1996

The broad bean gene VfNOD32 encodes a nodulin with sequence similarities to chitinases that is homologous to (alpha/beta)8-barrel-type seed proteins.

Nodulin gene transcripts isolated from a broad bean (Vicia faba L.) root nodule cDNA library and designated VfNOD32 are detectable in the nitrogen-fixing zone III of nodules and in much smaller amounts in flowers. In nodules, these transcripts are detectable for the first time 7 d after inoculation, at least 1 d before leghemoglobin gene transcription starts. Two putative full-length cDNAs representing different transcript sequences of 92.5% identity were sequenced. The corresponding broad bean genes were termed VfNOD32-A1 and VfNOD32-A2, and the encoded proteins were termed Nvf32-A1 and Nvf32-A2. The derived amino acid sequences of the Nvf32 proteins are highly homologous to the Vicia narbonensis (alpha/beta)8-barrel seed protein narbonin. Considering this homology, Nvf32 is assumed to have a similar structure consisting of beta-sheets forming a central barrel surrounded by alpha-helices. The two Nvf32 sequences also contain two conserved amino acid motifs that are characteristic of class-III chitinases. Several amino acids demonstrated to be essential for chitinase activity are conserved in both regions, whereas one essential glutamic acid was changed to glycine in the Nvf32-A1 isoform but not in the Nvf32-A2 isoform.

Amino Acid Sequence↗
PubMed2006

Adsorption at the air-water interface and emulsification properties of grain legume protein derivatives from pea and broad bean.

Functional properties of native and modified (through induced autolysis) pea (Pisum sativum L.) and broad bean (Vicia faba L.) protein derivatives are studied. In specific, protein solubility and behavior at the air-water interface through surface pressure measurements are investigated. Furthermore the ability of the protein products to act as emulsifying agents and to stabilize emulsions is studied through oil droplet size distribution measurements and by the protein adsorbed at the oil-water interface. The data reveal that the ability of the proteins to act as surfactants and build up a rigid film around the oil droplets, mainly depends on their suitable molecular configuration and structure. Hydrolysis did not promote the functionality of the legume proteins. Broad bean exhibited better functionality than pea, before and after hydrolysis. Some comparisons were also made with lupin (Lupinus albus L.) protein isolate.

Adsorption↗
PubMed1995

Immunostaining and interphase arrangement of field bean kinetochores.

More than 100 sera from patients with scleroderma CREST (calcinosis, Raynaud phenomenon, esophageal dismotility, sclerodactyly, telangiectasia) were tested in order to detect antigenic nuclear components of the field bean Vicia faba (2n = 12). Kinetochores of mitotic chromosomes and prekinetochores of interphase cells from root-tip meristems were specifically labelled via an indirect immunofluorescence procedure by antibodies of one of these sera. In 44% of interphase nuclei in which centromeres could be identified, only half (6) of the number of expected prekinetochores (12) was detected, circumstantially indicating at least transient association of homologous centromeres. Some nuclei showed clustering of centromeres at one pole (Rabl configuration). In metaphase chromosomes, each sister kinetochore contained a fluorescent spot. Western blotting of field bean nuclear proteins revealed four antigenic proteins of 28, 30, 64 and 68 kDa.

Blotting, Western↗
PubMed2005

Phytotoxic, clastogenic and bioaccumulation effects of the environmental endocrine disruptor bisphenol A in various crops grown hydroponically.

The effects of the endocrine disruptor bisphenol A (BPA) at concentrations of 10 and 50 mg l(-1) were evaluated on the germination and morphology, micronuclei (MN) content in root tip cells and BPA bioaccumulation of hydroponic seedlings of broad bean (Vicia faba L.), tomato (Lycopersicon esculentum Mill.), durum wheat (Triticum durum Desf.) and lettuce (Lactuca sativa L.) after 6 and 21 days of growth. In general, BPA at any dose used did not inhibit germination and early growth (6 days) of seedlings of the species examined, with the exception of primary root length of tomato which decreased at the higher BPA dose. In contrast, an evident phytotoxicity was induced by BPA in all species after 21 days of growth with evident morphological anomalies and significant reductions of the lengths and fresh and dry weights of shoots and roots of seedlings. With respect to the nutrient medium without seedlings, BPA concentration decreased markedly during the growth period in the presence of broad bean and tomato seedlings, and limitedly in the presence of durum wheat and, especially, lettuce. Further, the presence of BPA measured in roots and shoots of broad bean and tomato after 21-day growth indicated that bioaccumulation of BPA had occurred. The number of MN in broad bean and durum wheat root tip cells increased markedly by treatment with BPA at both concentrations, thus suggesting a potential clastogenic activity of BPA in these species.

Benzhydryl Compounds↗
PubMed1997

Formation and repair of O6-methylguanine in recombination hot spots of plant chromosomes.

Mutagen-induced chromatid aberrations are not randomly distributed along the metaphase chromosomes. In the field bean (Vicia faba), defined late-replicating and transcriptionally inactive heterochromatic regions are preferentially involved. After exposure to the alkylating agent N-methyl-N-nitrosourea (MNU) (10(-3) M, 1 hour), 70% of all aberrations are clustered within 6 segments containing tandemly repeated FokI elements of 59 bp, which comprise approximately 10% of the genome. Using immuno-slot-blot analyses, we have studied the frequency of O6-methylguanine (O6-MeG), a mutagenic lesion important for aberration induction, in total genomic DNA as well as in FokI sequences of the field bean after exposure to MNU. In either case, similar numbers of adducts per nucleotide were found immediately after treatment as well as after 18 hours of recovery, when most adducts were removed and significant amounts of chromatid aberrations were detectable. Peculiarities of long FokI element arrays (e.g., formation of specific tertiary structures), resulting in error-prone recombination repair, rather than preferential formation or delayed repair of O6-MeG are apparently responsible for aberration clustering in these hot spot regions.

Alkylating Agents↗
PubMed1988

Nutritional and technological characteristics of new broad bean flaked products.

The effects of the technological processes (soaking in water or alkaline solutions, drying, puree preparation) and the supplementation with maize flour on the nutritional value and on the organoleptic characteristics of broad bean (Vicia faba, L. major) flakes have been studied. Protein content was not affected by technological process. The addition of maize flour decreased the protein content of the final product depending on the amount of the maize flour added. Amino acid composition showed a decrease of tryptophan due to technological processing. Supplementation with maize flour improved the amino acid pattern and, except for tryptophan, the amount of essential amino acids in the flakes supplemented with 25% or more maize flour well compared with the provisional pattern by F.A.O. In vitro digestibility trials did not evidence significant changes due to technological processes or to integration of broad beans with maize flour. Broad bean toxic factors (vicine and convicine glycosides) were only slightly affected by the alkaline treatment of the flakes. Glycosides content decreased with the increasing supplementation with maize flour but the relationship was not linear. The organoleptic tests were positive for texture and taste, whereas the appearance of the products should be improved.

Amino Acids↗
PubMed1997

Partial purification of mitochondrial ribosomes from broad bean and identification of proteins encoded by the mitochondrial genome.

An approach towards the identification at the protein level of the ribosomal proteins encoded by the mitochondrial genome of broad bean (Vicia faba) has been developed. After Triton X-100 treatment of isolated mitochondria, a fraction enriched in mitochondrial ribosomes was obtained by successive centrifugation, first onto a sucrose cushion, and then in a sucrose gradient. Mitochondrial translation products were labelled in isolated mitochondria with [35S]methionine and added to the enriched mitochondrial ribosomal proteins before separation by two-dimensional gel electrophoresis. Six spots, identified both by Coomassie blue staining and autoradiography, were analysed by protein micro-sequencing. Two of these were shown to correspond to ribosomal proteins S10 and S12. We conclude that these two proteins are encoded by the mitochondrial genome of broad bean and that the method described here can be used to identify other proteins encoded by the mitochondrial genome.

Amino Acid Sequence↗
PubMed1987

Plant small nuclear RNAs. II. U6 RNA and a 4.5SI-like RNA are present in plant nuclei.

Two small nuclear RNA species (U6 RNA and a 4.5SI-like RNA) not described so far for plants were detected in broad bean (Vicia faba L.) nuclei. U6 RNA is 98 nucleotides long, contains psi and methylated nucleotides and shows a surprisingly high degree of sequence homology (80%) with its rat counterpart, particularly in the middle part (a 57 nucleotide-long stretch) of the molecule, where it amounts to 98%. The 4.5SI-like RNA, similar in its structure to 4.5SI RNA detected so far only in rodent nuclei, is 94 nucleotides long, contains psi and an unidentified nucleotide and exhibits 52% overall sequence homology with rat 4.5SI RNA. A block of 20 consecutive nucleotides at the 5' end of the molecule is conserved between broad bean 4.5SI-like RNA and rat 4.5SI RNA. The presence of the two RNA polymerase III internal promoter consensus sequences in 4.5SI-like RNA suggests that it is an RNA polymerase III transcript.

Animals↗
PubMed1991

Subrepeats of rDNA intergenic spacer present as prominent independent satellite DNA in Vigna radiata but not in Vigna angularis.

Subrepeats located in the rDNA intergenic spacer are also present as independently occurring, tandemly arranged satellite DNA clusters in the genome of Vigna radiata (mung bean). These 174-bp satellite repeats are identified as non-rDNA repeats by the presence of an AluI site. In the closely related Vigna angularis (adzuki bean), 174-bp repeats characterized by an AluI site occur in the rDNA with high sequence homology to the V. radiata rDNA subrepeats. A part of the 174-bp element that shows high similarity to a Xenopus terminator box (T2/T3) is slightly modified in V. angularis. However, a characteristic stem-loop structure can be formed, as in the case of V. radiata. Two highly conserved 12-bp regions occur within the 174-bp rDNA repeats of the two plants investigated. One of these 12-bp stretches exhibits some sequence identity to an element repeated twice in the 325-bp repeats in the intergenic spacer region of Vicia faba (broad bean).

Animals↗
PubMed1983

The alpha subunit of a plant mitochondrial F1-ATPase is translated in mitochondria.

The mitochondrial F1-ATPase from bean (Vicia faba L.) was solubilized by a chloroform treatment of mitochondrial membranes and purified by centrifugation on a glycerol gradient. The active fraction contained 5 subunits: alpha (Mr = 52,000), beta (Mr = 51,000), gamma (Mr = 34,000), delta (Mr = 23,800), and epsilon (Mr = 22,900). Purified coupled mitochondria were incubated in the presence of [ 35S ]methionine and malate to allow mitochondrial translation to occur. The largest labeled polypeptide (Mr = 52,000) was present in the chloroform extract, co-sedimented with the F1-ATPase on glycerol gradient and co-migrated with the alpha subunit upon two-dimensional electrophoresis. The results indicate that the alpha subunit of bean mitochondrial ATPase is translated on mitoribosomes, in contrast to the situation in other organisms.

Electrophoresis, Polyacrylamide Gel↗
PubMed2001

Carrier-mediated uptake and phloem systemy of a 350-Dalton chlorinated xenobiotic with an alpha-amino acid function.

In a previous paper we have shown that epsilon-(phenoxyalkanecarboxylyl)-L-Lys conjugates are potent inhibitors of amino acid transport systems and that it is possible to modulate the uptake inhibition by hydrophobic or hydrophilic additions in the 4-position of the aromatic ring (J.F. Chollet, C. Delétage, M. Faucher, L. Miginiac, J.L. Bonnemain [1997] Biochem Biophys Acta 1336: 331-341). In this report we demonstrate that epsilon-(2,4-dichlorophenoxyacetyl)-L-Lys (2,4D-Lys), one of the largest molecules of the series and one of the most potent inhibitors, is a highly permeant conjugate. Uptake of 2,4D-Lys by broad bean (Vicia faba) leaf discs is mediated by an active carrier system (Km1 = 0.2 mM; Vmax1 = 2.4 nmol x cm(-2) x h(-1) at pH 5.0) complemented by an important diffusive component. Among the compounds tested (neutral, basic, and acidic amino acids, auxin, glutathione, and sugars), only the aromatic amino acids clearly compete with 2,4D-Lys. The conjugate accumulates in the vein network, is exported toward the growing organs, and exhibits a distribution pattern different from that of the herbicide moiety. However, over time 2,4D-Lys progressively splits into 2,4D and lysine. Analyses by high-performance liquid chromatography and liquid scintillation spectrometry of the phloem sap collected from the castor bean system, used as a systemy test, indicate decreasing capacities of 2,4D, 2,4D-Lys, and glyphosate, respectively, to move from the epidermis cell wall to the sieve element. Our results show that it is possible to design synthesis of large-size xenobiotics (approximately 350 D) with a lipophilic pole, exhibiting high mobility within the vascular system.

4-Chloromercuribenzenesulfonate↗
PubMed2000

Laser pretreatment protects cells of broad bean from UV-B radiation damage.

In order to determine the role of lasers in the stress resistance of broad bean (Vicia faba L.) to ultraviolet-B (UV-B) radiation, the embryos in seeds were exposed to He-Ne laser or CO2 laser radiation. Afterwards they were cultivated in Petri dishes in a constant temperature incubator until the lengths of epicotyls were nearly 3 cm. The epicotyls were then exposed to 1.02, 3.03 or 4.52 kJ m(-2) UV-B radiation, respectively, under 70 micromol m(-2) s(-1) photosynthetically active radiation (PAR) in a growth cabinet. Changes in the concentration of malondialdehyde (MDA), ascorbic acid (AsA) and UV-B absorbing compounds (absorbance at 300 nm) were measured to test the effects of laser pretreatment. The results showed that laser pretreatment of embryos enhanced UV-B stress resistance in the epicotyls of the broad bean by decreasing the MDA concentration and increasing the content of AsA and UV-B absorbing compounds. We suggest that those changes in MDA, AsA and UV-B absorbing compounds were responsible for the increase in stress resistance observed in the broad bean. This is the first investigation reporting the use of laser pretreatment to protect the cells of the broad bean from UV-B-induced damage.

Ascorbic Acid↗
PubMed1998

Detection of Ca2+-dependent transglutaminase activity in root and leaf tissue of monocotyledonous and dicotyledonous plants

Protein extracted from root and leaf tissue of the dicotyledonous plants pea (Pisum sativum) and broad bean (Vicia faba) and the monocotyledonous plants wheat (Triticum aestivum) and barley (Hordeum vulgare) were shown to catalyze the incorporation of biotin-labeled cadaverine into microtiter-plate-bound N', N'-dimethylcasein and the cross-linking of biotin-labeled casein to microtiter-plate-bound casein in a Ca2+-dependent manner. The cross-linking of biotinylated casein and the incorporation of biotin-labeled cadaverine into N',N'-dimethylcasein were time-dependent reactions with a pH optimum of 7.9. Transglutaminase activity was shown to increase over a 2-week growth period in both the roots and leaves of pea. The product of transglutaminase's protein-cross-linking activity, epsilon-(gamma-glutamyl)-lysine isodipeptide, was detected in root and shoot protein from pea, broad bean, wheat, and barley by cation-exchange chromatography. The presence of the isodipeptide was confirmed by reversed-phase chromatography. Hydrolysis of the isodipeptide after cation-exchange chromatography confirmed the presence of glutamate and lysine.

Journal Article↗
PubMed1996

Loss of particulate contaminants from plant canopies under wet and dry conditions.

There is a requirement for data describing the loss with time of particulate contamination from plant canopies. Measurements were made of the loss rates of monodispersed silica spheres (three sizes, with Mass Median Aerodynamic Diameters (MMADs) 1.9, 5.3 and 8.4 microm) from wheat (Triticum aestivum) and broad bean (Vicia faba) canopies. The spheres were labelled with tracers detectable by Instrumental Neutron Activation Analysis (INAA). Canopies were contaminated under realistic turbulence conditions in a wind tunnel, then removed to sheltered and exposed field sites or to a glasshouse containing a rain simulator. Samples were taken periodically, and the level of contamination per plant determined by INAA. Statistical analysis of the resulting data suggested an offset exponential loss model, with a residue of deposit that is not lost over time. Loss half-lives in the order of 1-2 days were obtained for an exposed wheat crop and 3-4 days for a partially sheltered wheat crop, with permanent residues of initial deposit for the exposed crop of 4-8%, and for the partially sheltered crop of 22-52%. A broad bean crop under glasshouse conditions showed loss half-lives of 0.5-1.5 days with residues of 22-26% initial contamination. A double exponential loss model also fitted the data well in some cases, and it is possible that a slow loss of the residual deposit occurs, being masked by noise in the current data set.

Journal Article↗
PubMed1992

Broad bean leaf polyphenol oxidase is a 60-kilodalton protein susceptible to proteolytic cleavage.

Polyphenol oxidase (PPO) in leaves is generally considered to be a 45-kilodalton protein. PPO purified to homogeneity from broad bean (Vicia faba L.) leaves in the presence of protease inhibitors had an apparent molecular mass of 60 kD determined by denaturing sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. Under partially denaturing conditions, the protein had an apparent molecular mass of 43 kilodaltons, but this was shifted to 60 kilodaltons in the presence of sulfhydryl reductants, suggesting the presence of disulfide bonding. The purified protein was totally latent; i.e. PPO activity was only detected when assayed with sodium dodecyl sulfate. Treatment with proteases in the presence of 0.1% SDS inhibited enzyme activity, but in the absence of SDS the 60-kilodalton PPO was proteolytically cleaved with no loss of PPO activity. This yielded a 42-kilodalton peptide that had PPO activity and inactive peptides of 12 to 18 kilodaltons. Amino acid sequencing established that the 42-kilodalton protein was derived from the N-terminal end of the 60-kilodalton form of PPO. By comparison with the sequence of a cDNA clone for broad bean leaf PPO, the 18-kilodalton peptide was located at the C-terminal end of the 60-kilodalton protein. Northern analysis of mRNA from broad bean leaves probed with a cDNA clone of PPO identified a transcript of 2.2 kilobase pairs, which is more than sufficient to encode the 60-kilodalton protein. It is concluded that PPO is a 60-kilodalton protein in broad bean leaves but that it is susceptible to proteolysis during extraction, removing a peptide of 15 to 18 kilodaltons from the C-terminal end without decreasing enzyme activity.

Journal Article↗
PubMedDate not supplied

Chemical analysis and hemolytic activity of the fava bean aglycon divicine.

Divicine is an unstable aglycon metabolite of the fava bean pyrimidine beta-glucoside vicine. Divicine has long been thought to be a mediator of an acute hemolytic crisis, known as favism, in susceptible individuals who ingest fava beans (Vicia faba). However, a recent report has questioned the chemical identity of the divicine that was used in most of the studies on divicine hemotoxicity. The present study was undertaken to examine the hemolytic potential of synthetic divicine. Divicine was synthesized and its identity and purity were confirmed by HPLC, mass spectrometry, and NMR spectroscopy. The stability and redox behavior of divicine, under physiological conditions, were examined by HPLC and cyclic voltammetry. The data indicate that divicine is readily oxidized under aerobic conditions; however, it was sufficiently stable at pH 7.4 to permit its experimental manipulation. When 51Cr-labeled rat erythrocytes were exposed in vitro to the parent glucoside, vicine (5 mM), and then readministered to rats, no decrease in erythrocyte survival was observed. In contrast, erythrocyte survival was dramatically reduced by in vitro exposure to divicine (1.5 mM). These data demonstrate that divicine is a direct-acting hemolytic agent and thus may be a mediator of the hemolytic crisis induced by fava bean ingestion.

Animals↗