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PubMed1985

Occurrence and detection of natural mutagens and modifying factors in food products.

Various food products of plant origin were investigated for the occurrence of natural mutagens using the Salmonella/microsome assay. In general, food plants were freeze-dried and subsequently extracted with a number of solvents. Solvents were evaporated and the residues obtained were tested for mutagenicity. In addition to S9-mix, gut flora extracts were applied for metabolic activation. From bracken fern (Pteridium aquilinum) a novel mutagen, designated Aquilide A, was isolated and its chemical structure was identified. Aquilide A requires activation to become mutagenic. This activation occurs spontaneously at pH levels above 6-7. Activated Aquilide A was found to be genotoxic in cultured mammalian cells. Natural mutagens were detected in 4 out of 6 vegetables investigated. In addition, broad beans (Vicia faba) were found to be mutagenic after treatment with nitrite. All mutagenic vegetables showed marked intercultivar variations. From lettuce and string beans quercetin was isolated (after chemical hydrolysis) and in rhubarb emodin, an anthraquinon, was detected. The mutagenic activity of these two compounds was further investigated using cultured mammalian cells. Quercetin and emodin responded negative or weakly positive in the systems applied. The genotoxic properties of a number of pyrrolizidin alkaloids, which are reported to occur in various flowering plants and as a result occur in honey and some herbal preparations, were studied using a cocultivation system of V79 Chinese hamster cells and primary cultures of chick embryo hepatocytes (PCCEH/V79). All four pyrrolizidine alkaloids investigated were found to be potent inducers of SCEs in this test system. Anti-mutagenic effects of butylated hydroxyanisole (BHA) and indole-3-carbinol (I3C) were detected using the PCCEH/V79 cocultivation system. This indicates that the cocultivation system described can be a valuable tool for the screening of various products for potential anti-carcinogenic properties. Extracts of lettuce and string beans, and a number of natural chemicals were found to reduce the mutagenic activity of cigarette smoke condensate and benzo (a) pyrene (BaP) as detected in the Salmonella/microsome assay. Intercultivar variation with respect to the antimutagenic activity observed was less pronounced than the variation noted for the mutagenic activity of these vegetables. Measures which may result in a reduction of the exposure to a number of natural mutagens are discussed.

Animals↗
PubMed1989

Photoinhibition at low temperature in chilling-sensitive and -resistant plants.

Photoinhibition resulting from exposure at 7 degrees C to a moderate photon flux density (300 micromoles per square meter per second, 400-700 nanometers) for 20 hours was measured in leaves of annual crops differing widely in chilling tolerance. The incidence of photoinhibition, determined as the decrease in the ratio of induced to total chlorophyll fluorescence emission at 693 nanometers (F(v)/F(max)) measured at 77 Kelvin, was not confined to chilling-sensitive species. The extent of photoinhibition in leaves of all chilling-resistant plants tested (barley [Hordeum vulgare L.], broad bean [Vicia faba L.], pea [Pisum sativum L.], and wheat [Triticum aestivum L.]) was about half of that measured in chilling-sensitive plants (bean [Phaseolus vulgaris L.], cucumber [Cucumis sativus L.], lablab [Lablab purpureus L.], maize [Zea mays L.], pearl millet [Pennisetum typhoides (Burm. f.) Stapf & Hubbard], pigeon pea [Cajanus cajun (L.) Millsp.], sesame [Sesamum indicum L.], sorghum [Sorghum bicolor L. Moench], and tomato [Lycopersicon esculentum Mill.]). Rice (Oryza sativa L.) leaves of the indica type were more susceptible to photoinhibition at 7 degrees C than leaves of the japonica type. Photoinhibition was dependent both on temperature and light, increasing nonlinearly with decreasing temperature and linearly with increasing light intensity. In contrast to photoinhibition during chilling, large differences, up to 166-fold, were found in the relative susceptibility of the different species to chilling injury in the dark. It was concluded that chilling temperatures increased the likelihood of photoinhibition in leaves of both chilling-sensitive and -resistant plants. Further, while the photoinhibition during chilling generally occurred more rapidly in chilling-sensitive plants, this was not related directly to chilling sensitivity.

Journal Article↗
PubMed1977

Phospholipid Biosynthesis and Fatty Acid Content in Relation to Chilling Injury during Germination of Seeds.

The proportion of labeled (14)C-glycerol incorporated into phospholipids and the fatty acid composition of three phospholipids in germinating seeds and seedlings of chilling-sensitive lima beans (Phaseolus lunatus L.) and chilling-resistant broad beans (Vicia faba L.) and peas (Pisum sativum L.) at 10 and 25 C were determined. During the imbibition of seeds (first 24 hours), lima beans were sensitive to chilling injury at 10 C and a higher proportion of label was incorporated into phosphatidylethanolamine and phosphatidylglycerol than in broad beans and peas. Broad beans and peas incorporated a higher proportion of label into phosphatidylcholine. The oleic acid content of phosphatidylcholine was higher and linolenic acid content was lower in peas and broad beans than in lima beans at 10 and 25 C. The unsaturated to saturated fatty acid ratio was much higher for the chilling-resistant seeds than for the chilling-sensitive ones. In the seedling stage, the proportion of label incorporated into the four major phospholipids was similar in the three species regardless of temperature treatment. The fatty acid content of the phospholipids examined was not different in the three species in the seedling stage.

Journal Article↗
PubMed2004

Diversity and specificity of Rhizobium leguminosarum biovar viciae on wild and cultivated legumes.

The symbiotic partnerships between legumes and their root-nodule bacteria (rhizobia) vary widely in their degree of specificity, but the underlying reasons are not understood. To assess the potential for host-range evolution, we have investigated microheterogeneity among the shared symbionts of a group of related legume species. Host specificity and genetic diversity were characterized for a soil population of Rhizobium leguminosarum biovar viciae (Rlv) sampled using six wild Vicia and Lathyrus species and the crop plants pea (Pisum sativum) and broad bean (Vicia faba). Genetic variation among 625 isolates was assessed by restriction fragment length polymorphism (RFLP) of loci on the chromosome (ribosomal gene spacer) and symbiosis plasmid (nodD region). Broad bean strongly favoured a particular symbiotic genotype that formed a distinct phylogenetic subgroup of Rlv nodulation genotypes but was associated with a range of chromosomal backgrounds. Host range tests of 80 isolates demonstrated that only 34% of isolates were able to nodulate V. faba. By contrast, 89% were able to nodulate all the local wild hosts tested, so high genetic diversity of the rhizobial population cannot be ascribed directly to the diversity of host species at the site. Overall the picture is of a population of symbionts that is diversified by plasmid transfer and shared fairly indiscriminately by local wild legume hosts. The crop species are less promiscuous in their interaction with symbionts than the wild legumes.

Base Sequence↗
PubMed2000

N-Methyl-N-nitrosourea-induced DNA damage detected by the comet assay in Vicia faba nuclei during all interphase stages is not restricted to chromatid aberration hot spots.

The genotoxic effect of the monofunctional alkylating agent N:-methyl-N:-nitrosourea (MNU) on root-tip nuclei of the field bean, Vicia faba, has been tested by comparative application of three protocols of the comet assay. While the alkaline denaturation/alkaline electrophoresis (A/A) procedure proved to be most sensitive at low doses, the alkaline denaturation/neutral electrophoresis (A/N) procedure yielded an optimal dose-response curve within a wider dose range. With the neutral electrophoresis without alkaline denaturation (N/N) procedure only minimal response was found. MNU-mediated single-strand breaks occurred in nuclei of all interphase stages. Detection of tandemly repeated FOK:I elements on comets by fluorescence in situ hybridization showed an average involvement of these heterochromatin-specific sequences in MNU-mediated single-strand breaks. This, together with previous results, suggests that the pronounced clustering of chromosomal aberrations in heterochromatic regions after treatment with S phase-dependent mutagens is mainly due to an error-prone interference of recombinative repair and replication in damaged basic repeats of large tandem repeat arrays.

Alkylating Agents↗
PubMed2002

A new peptidic protease inhibitor from Vicia faba seeds exhibits antifungal, HIV-1 reverse transcriptase inhibiting and mitogenic activities.

A new trypsin-chymotrypsin Inhibitor, with an N-terminal sequence showing some differences from the previously reported trypsin-chymotrypsin inhibitor, was isolated from the broad bean Vicia faba. The inhibitor was a peptide with a molecular mass of 13 kDa. It was adsorbed on Affi-gel blue gel and CM-Sepharose. It exerted antifungal activity toward Mycosphaerella arachidicola and Physalospora piricola. In addition, the trypsin-chymotrypsin inhibitor elicited a mitogenic response from mouse splenocytes and inhibited the activity of human immunodeficiency virus-1 reverse transcriptase.

Amino Acid Sequence↗
PubMed2005

Effects of magnesium availability on the activity of plasma membrane ion transporters and light-induced responses from broad bean leaf mesophyll.

Considering the physiological significance of Mg homeostasis in plants, surprisingly little is known about the molecular and ionic mechanisms mediating Mg transport across the plasma membrane and the impact of Mg availability on transport processes at the plasmalemma. In this study, a non-invasive ion-selective microelectrode technique (MIFE) was used to characterize the effects of Mg availability on the activity of plasma membrane H+, K+, Ca2+, and Mg2+ transporters in the mesophyll cells of broad bean (Vicia faba L.) plants. Based on the stoichiometry of ion-flux changes and results of pharmacological experiments, we suggest that at least two mechanisms are involved in Mg2+ uptake across the plasma membrane of bean mesophyll cells. One of them is a non-selective cation channel, also permeable to K+ and Ca2+. The other mechanism, operating at concentrations below 30 microM, was speculated to be an H+/Mg+ exchanger. Experiments performed on leaves grown at different levels of Mg availability (from deficient to excessive) showed that Mg availability has a significant impact on the activity of plasma-membrane transporters for Ca2+, K+, and H+. We discuss the physiological significance of Mg-induced changes in leaf electrophysiological responses to light and the ionic mechanisms underlying this process.

Biological Transport↗
PubMed1991

In situ evaluation of the ruminal and intestinal degradability of extruded whole horse beans.

Four rumen and proximal duodenum fistulated non-lactating Holstein cows were used to determine the effect of extrusion at 120 degrees C of whole horse beans (Vicia faba cv Talo) on in vitro nitrogen (N) solubility and in situ degradation of dry matter (DM) and crude protein (CP) in the rumen and intestine. Cows were fed a ration of 30% whole horse beans (WHB) and 70% Italian rye-grass hay. The degradation of DM and CP was estimated using nylon bags suspended in the rumen for 2, 4, 7, 16, 24 and 48 h; the effective ruminal degradability of DM and CP was evaluated assuming a ruminal outflow rate of 0.06/h. Bags incubated in the rumen for 16 h were introduced into the small intestine through the duodenal cannula and subsequently recovered in the feces. Extrusion of WHB reduced N-solubility in buffer solution (21.1 vs 74.9%). Processing diminished the effective rumen degradability of DM (74.6 vs 80.4%) and CP (70.2 vs 89.2%). Meanwhile, the amounts of DM and CP digested in the intestine increased: 9.6 vs 1.4% and 25.2 vs 3.0% respectively. Therefore, feeds containing extruded WHB increase the availability of dietary proteins in the intestine compared with diets containing raw WHB.

Animal Feed↗
PubMed1962

Synthetic capacitie of chromosome fragments correlated with their ability to maintain nucleolar material.

Onion (Allium cepa) and bean (Vicia faba) root tip cells containing many micronuclei, derived from x-ray-induced chromosome fragments, were exposed to H(3)-thymidine and H(3)-cytidine to determine the ability of such fragments to undergo DNA and RNA synthesis. Only a few micronuclei in onion and many in bean roots synthesize nucleic acid simultaneously with their main nuclei. A few micronuclei labeled with H(3)-thymidine undergo mitotic chromosome condensation along with the main nuclei, while the unlabeled ones never do so. The onset of nucleic acid synthesis as well as mitosis in micronuclei appears to be under generalized cellular control. Although all chromosomes and chromosome fragments at telophase give a positive reaction for a silver stainable nucleolar fraction, in the subsequent interphase only some micronuclei, derived from such chromosome fragments, are found to maintain nucleoli; others lose them with time. Those micronuclei which maintain nucleoli, perhaps due to the presence of specific chromosomal regions, are also active in DNA and RNA synthesis. These results are compatible with the concept that nucleoli and associated chromosome regions play an important role in the primary biosynthetic processes of the cell.

Cell Nucleolus↗
PubMed1996

Tomographic distribution of acetylated histone H4 in plant chromosomes, nuclei and nucleoli.

Root tip cells of broad bean (Vicia faba L. cv. 'Wase soramame') and barley (Hordeum vulgare L. cv. 'Minorimugi') were immunostained with antibodies specific for acetylated histone H4. With an antiserum that recognizes histone H4 acetylated at lysine-5, the nucleolar organizing region (NOR) in mitotic chromosomes was strongly labeled in both species. The broad bean had two signals in the metaphase and telophase chromosome complements and four signals in the prophase and anaphase chromosome complements, while the barley had four signals in the metaphase and telophase chromosome complements and eight signals in the prophase and anaphase complements. Five different patterns of signals were observed at interphase: in type I only nucleoli were wholly stained; in type II perinucleolar knob-like signals and/or fiber-like signals emanated from the nucleus; in type III aggregate signals appeared in the nucleolus; in type IV many small dot-like signals were distributed throughout the nucleus, except nucleoli; and in type V string-like or some granule-like signals appeared in the nucleoli. Type II was very similar to previous results by in situ hybridization with sense rDNA probes. Type III was similar to the patterns of DNA synthesis recognized as chromatin domains by anti-BrdU antibodies. Type V was very similar to the results of in situ hybridization with pTa71, rDNA probes and the appearance of the dense fibrillar components of the nucleolus.

Acetylation↗
PubMed1988

Ribosomal protein S14 genes in broad bean mitochondrial DNA.

Broad bean (Vicia faba) mtDNA contains an open reading frame with a predicted amino acid sequence that is 41% homologous to the ribosomal protein S14 (RPS14) of Escherichia coli, and which is located 1232 ntp upstream from a gene for cytochrome b (cob). A second putative rpS14 gene occurs in broad bean mtDNA, 344 ntp upstream from a gene for ATPase subunit 9 (atp9). However, the atp9-linked rpS14 gene is 12 codons shorter than the cob-linked rpS14 gene. Sequence homology is found upstream (for 218 ntp) but not downstream from the two rpS14 genes. Transcripts were detected in broad bean mtRNA only for the cob-linked rpS14 gene. All RNA molecules that include a transcript of the rpS14 gene also include a transcript of the cob gene. Sequences homologous to the broad bean mitochondrial rpS14 gene were detected in soybean mtDNA, but not in corn mtDNA. Relationships between the amino acid sequences of RPS14s encoded in broad bean mtDNA, in chloroplast DNAs of various angiosperms, and in E. coli are consistent with the view that the ancestral lines of these three kinds of DNA diverged from each other within a relatively short time period.

Amino Acid Sequence↗
PubMed1983

Secretion of plant storage globulin polypeptides by Xenopus laevis oocytes.

Xenopus oocytes injected with poly(A)-containing RNA from developing cotyledons of field beans (Vicia faba L. var. minor) synthesize precursor polypeptides to the major storage globulins legumin and vicilin. These polypeptides are secreted into the medium without proteolytic cleavage of the legumin propolypeptides into the mature disulfide-linked alpha and beta chains. Similarly, storage globulin polypeptides from pea (Pisum sativum L.) and french bean (Phaseolus vulgaris L.) were secreted from oocytes. Inhibition of glycosylation by tunicamycin does not prevent secretion. This first report on the secretion of plant polypeptides by Xenopus oocytes shows that (a) intracellular deposition of storage proteins in membrane-bounded organelles (protein bodies) of plants and extracellular secretion have step(s) in common, and (b) the cell, in addition to the mRNA, determines the final destination of these polypeptides.

Animals↗
PubMed2006

VfCPK1, a gene encoding calcium-dependent protein kinase from Vicia faba, is induced by drought and abscisic acid.

Calcium, one of the most ubiquitous second messengers, has been shown to be involved in a wide variety of responses in plants. Calcium-dependent protein kinases (CDPKs) (EC 2.7.1.37) are the predominant Ca(2+)-regulated serine/threonine protein kinase in plants and play an important role in plant calcium signal transduction. CDPKs are encoded by a large multigene family in many plants, which has been showed so far; however, the precise role of each specific CDPK is still largely unknown. A novel CDPK gene designated as VfCPK1 was cloned from epidermal peels of broad bean (Vicia faba L.) leaves using the rapid amplification of cDNA ends (RACE)-PCR technique and its expression was studied in detail. The VfCPK1 cDNA is 1783 bp long and contains an open reading frame of 1482 bp encoding 493 amino acids. VfCPK1 contains all conserved regions found in CDPKs and shows a high level of sequence similarity to many other plant CDPKs. VfCPK1 was highly expressed in leaves, especially in leaf epidermal peels of broad bean in mRNA and protein levels. Expressions of VfCPK1 at both the mRNA and protein levels were increased in leaves treated with abscisic acid or subjected to drought stress. Potential roles of VfCPK1 in epidermal peels are discussed. The nucleotide sequence data reported here were deposited in the GenBank database under accession number AY753552.

Abscisic Acid↗
PubMed1982

[Two forms of carbonic anhydrase from bean chloroplasts].

The intracellular localization of carboanhydrase from the bean (Vicia faba) leaves was investigated. The bulk of the enzyme activity (about 70%) is localized in the chloroplasts. The enzyme exists both in the soluble and in membrane-bound state. Disc electrophoresis in polyacrylamide gel with specific staining for the enzyme activity demonstrated that the soluble and membrane-bound carboanhydrases from bean chloroplasts are two molecular forms (presumably isoenzymes) differing in their electrophoretic mobilities. Protein fractionation with (NH4)2SO4 and a subsequent gel filtration on Sephadex G-200 and isoelectrofocusing resulted in separation of the two forms and their purification to homogeneity. Both carboanhydrases differ in pH, specific activity and stability.

Carbonic Anhydrases↗
PubMed1980

Interactions between Rhizobia and Lectins of Lentil, Pea, Broad Bean, and Jackbean.

A quantitative method was developed to measure the binding of fluorescent-labeled lentil (Lens esculenta Moench), pea (Pisum sativum L.), broad bean (Vicia faba L.), and jackbean (Canavalia ensiformis L., DC.) lectins to various Rhizobium strains. Lentil lectin bound to three of the five Rhizobium leguminosarum strains tested. The number of lentil lectin molecules bound per R. leguminosarum 128C53 cell was 2.1 x 10(4). Lentil lectin also bound to R. japonicum 61A133. Pea and broad bean lectins bound to only two of the five strains of R. leguminosarum, whereas concanavalin A (jackbean lectin) bound to all strains of R. leguminosarum, R. phaseoli, R. japonicum, and R. sp. tested. Since these four lectins have similar sugarbinding properties but different physical properties, the variation in bindings of these lectins to various Rhizobium strains indicates that binding of lectin to Rhizobium is determined not only by the sugar specificity of the lectin but also by its physical characteristics.The binding of lentil lectin and concanavalin A to R. leguminosarum 128C53 could be inhibited by glucose, fructose, and mannose. However, even at 150 millimolar glucose, about 15% of the binding remained. The binding of lentil lectin to R. japonicum 61A133 could be inhibited by glucose but not by galactose. It is concluded that the binding site of lentil lectin to R. japonicum is different from the binding site of soybean lectin to R. japonicum.

Journal Article↗
PubMed2004

Thermodynamic and functional properties of legumin (11S globulin from Vicia faba) in the presence of small-molecule surfactants: effect of temperature and pH.

We report on the effect of a set of water-dispersible small-molecule surfactants (the main and the longest-hydrocarbon components of which are a citric acid ester of monostearate, a sodium salt of stearol-lactoyl lactic acid, and a polyglycerol ester of stearic acid) on molecular, thermodynamic, and functional properties of the major storage protein of broad beans (Vicia faba) legumin in different molecular states (native, heated, and acid-denatured). The interaction between legumin and the surfactants has been characterized by a combination of thermodynamic methods, namely, mixing calorimetry and multiangle laser static and dynamic light scattering. It was found that hydrogen bonds, electrostatic interactions, and hydrophobic contacts provided a basis for the interactions between the surfactants and both the native and the denatured protein in aqueous medium. Intensive association of the protein molecules in a bulk aqueous medium in the presence of the surfactants was revealed by static and dynamic laser light scattering. In consequence of this, both the surface activity and the gel-forming ability of legumin increased markedly, which has been shown by tensiometry, estimation of protein foaming capacity, and steady-state viscometry. A likely molecular mechanism underlying the effects of small-molecule surfactants on legumin structure-forming properties at the interface and in a bulk aqueous medium is discussed.

Acids↗
PubMed1981

[Carboanhydrase from bean leaves].

A highly purified preparation of carboanhydrase from the leaves of the bean Vicia faba var. major (Harz) F. Janthina was obtained. The enzyme was homogeneous during analytical disc-electrophoresis in polyacrylamide gel and analytical ultracentrifugation. The molecular weight of the enzyme is 270,000. The enzyme molecule contains six Zn atoms, has a quaternary structure and is made up of six subunits with molecular weights of 45,000. Some properties of the enzymes from bean leaves and pea leaves were compared. The enzymes differ in molecular weights and behaviour during DEAE-cellulose chromatography and gel filtration through Sephadex G-200.

Carbonic Anhydrases↗