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PubMed1992

A broad bean cDNA clone encoding a DNA-binding protein resembling mammalian CREB in its sequence specificity and DNA methylation sensitivity.

A plant cDNA has been cloned that encodes a DNA-binding protein displaying a nucleotide (nt) sequence specificity similar to that of the mammalian cyclic AMP response element-binding protein/activating transcription factor (CREB/ATF) family of mammalian proteins. This cDNA was cloned in Escherichia coli from a broad bean (Vicia faba) cDNA expression library using a recognition site probe. The deduced amino acid (aa) sequence of the recombinant cDNA-encoded protein, called VBP1, has a basic region adjacent to a leucine zipper motif, of the type seen in the DNA-binding domains of many eukaryotic DNA-binding proteins, including mammalian CREB/ATF. Although this aa sequence has homology to regions of deduced aa sequences of other cloned plant cDNAs, it is distinct in both the derived primary structure and in its nt sequence specificity. VBP1, as well as proteins in nuclear extracts of V. faba with similar nt sequence specificity, have their binding to DNA suppressed more than tenfold by cytosine methylation at the CREB/ATF consensus sequence.

Amino Acid Sequence↗
PubMed2004

Study of temperature influence on electron transport in higher plants via delayed luminescence method: experiment, theory.

The temperature dependence of steady-state millisecond delayed luminescence (DL) is studied within the temperature range from -23 to 45 degrees C in leaf segments of chinese rose (Hibiscus rosa-sinensis L.) and bean (Ficia faba). To describe the experimental dependence of DL steady-state intensity on temperature theoretically, we suggest the temperature dependences of rate constants in earlier proposed model of photosynthesis. Under these conditions, the temperature dependence of DL steady-state value has the same form as experimental curve.

Biosensing Techniques↗
PubMed2005

Assessment of the genotoxicity of 137Cs radiation using Vicia-micronucleus, Tradescantia-micronucleus and Tradescantia-stamen-hair mutation bioassays.

Since the middle of the 20th century, ionizing radiations from radioactive isotopes including 137Cs have been investigated to determine their genotoxic impact on living organisms. The present study was designed to compare the effectiveness of three plant bioassays to assess DNA damage induced by low doses of 137Cs: Vicia-micronucleus test (Vicia-MCN), Tradescantia-micronucleus test (Trad-MCN) and Tradescantia-stamen-hair mutation test (Trad-SH) were used. Vicia faba (broad bean) and Tradescantia clone 4430 (spiderwort) were exposed to 137Cs according to different scenarios: external and internal (contamination) irradiations. Experiments were conducted with various levels of radioactivity in solution or in soil, using solid or liquid 137Cs sources. The three bioassays showed different sensitivities to the treatments. Trad-MCN appeared to be the most sensitive test (significative response from 1.5 kBq/200 ml after 30 h of contamination). Moreover, at comparable doses, internal irradiations led to larger effects for the three bioassays. These bioassays are effective tests for assessing the genotoxic effects of radioactive 137Cs pollution.

Biological Assay↗
PubMed1999

Assessment of the genotoxicity of contaminated soil with the Allium/Vicia-micronucleus and the Tradescantia-micronucleus assays.

The present study concerns the genotoxicity of contaminated soil near Metz, France. Three plant bioassays, the Vicia faba (broad bean), the Allium cepa (white onion) and the Tradescantia (spiderwort) micronuclei tests were used to evaluate for genotoxicity. Two soil samples were tested: soil sample A (from an industrial waste site) and soil sample B (from a cokeworks waste site). Maleic hydrazide was used as the positive control. Aqueous extracts of the soil samples were used to treat the roots of Vicia and Allium, and plant cuttings of Tradescantia according to the standard protocol for these plant assays established by the International Program on Chemical Safety and the World Health Organization. The results of these tests showed differential sensitivity in the three different bioassays. Soil sample A was more toxic than soil sample B.

Allium↗
PubMed1996

The alkaline comet test on plant cells: a new genotoxicity test for DNA strand breaks in Vicia faba root cells.

The alkaline comet assay (single cell gel electrophoresis assay) is a sensitive method for the detection of DNA damage. This paper describes the first application of this assay to plant cells for genotoxicological assessment. Germinating Vicia faba (field bean) seedlings were kept in water with either methyl methanesulfonate (MMS), ethyl methanesulfonate (EMS), mitomycin C (MMC), cycloheximide (CH), cadmium chloride (CdCl2), potassium dichromate (K2Cr2O7), or chromium trichloride (CrCl3). Nuclei were isolated from the root cells and evaluated for the extent of DNA migration. With the exception of cycloheximide, all agents induced a significant increase in DNA migration. These results indicate that the comet assay may be a valuable tool for monitoring DNA damage in plant systems. However, there was a significant heterogeneity in the extent of DNA migration within and between seedlings, which may be intrinsic to the assay or indicative of sampling problems.

Cadmium↗
PubMed1999

Radiocaesium and radiostrontium uptake by turnips and broad beans via leaf and root absorption.

One of the immediate consequences of massive radioisotope release into the atmosphere is contamination of the biosphere. This contamination can affect plants either by direct deposition onto the leaves, or by contaminating the soil followed by absorption by the roots. Knowledge of the efficacy of the two routes of radionuclide incorporation into the food chain is fundamental to understanding the mechanisms by which radioactive contamination reaches man. The present work analyzes the incorporation of 134Cs and 85Sr via root and leaf uptake into the parts consumed by man, for two very different crops: turnip (Brassica napus) and broad bean (Vicia faba). The root uptake studies consider the available soil fraction for these two radionuclides, and indicate greater availability for 85Sr than for 134Cs which is fixed rapidly in the soil. For the study of leaf uptake, leaves were contaminated at three different stages of plant growth; the results indicate an inverse dependence of the transfer coefficients on the time elapsed from the moment of the contamination to harvesting of the edible parts.

Air Pollutants, Radioactive↗
PubMed1998

Photoinhibition of photosynthesis in vivo results in singlet oxygen production detection via nitroxide-induced fluorescence quenching in broad bean leaves.

In plants experiencing environmental stress, the formation of reactive oxygen is often presumed. In this study, singlet oxygen was detected in broad bean (Vicia faba) leaves that were photoinhibited in vivo. Detection was based on the reaction of singlet oxygen with DanePy (dansyl-2,2,5,5-tetramethyl-2,5-dihydro-1H-pyrrole) yielding a nitroxide radical (DanePyO) which is EPR active and also features lower fluorescence compared to DanePy. The two (fluorescent and spin) sensor fuctions of DanePy are commensurate, which makes detecting singlet oxygen possible with a spectrofluorimeter in samples hard to measure with EPR spectroscopy [Kálai, T., Hideg, E., Vass, I., and Hideg, K. (1998) Free Radical Biol. Med. 24, 649-652]. We found that in leaves saturated with DanePy, the fluorescence of this double sensor was decreased when the leaves were photoinhibited by 1500 micromol m-2 s-1 photosynthetically active radiation. This fluorescence quenching is the first direct experimental evidence that photoinhibition of photosynthesis in vivo is accompanied by 1O2 production and is, at least partly, governed by the process characterized as acceptor side-induced photoinhibition in vitro.

Dansyl Compounds↗
PubMed1998

Scavenging Effect of Water Soluble Proteins in Broad Beans on Free Radicals and Active Oxygen Species.

Water soluble proteins (WSP) in broad beans, Vicia faba, were purified, and their scavenging effects on free radicals and active oxygen species were investigated. The purification steps included ammonium sulfate precipitation followed by sequential chromatography on Sephadex G-75. The final gel filtration step yielded two peaks of scavenging activity, each containing M(r) values of 70 kDa (peak I) and 28 kDa (peak II). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of peak II fraction gave a single band with M(r) of 14 kDa, indicating that peak II protein is dimeric. WSP exhibited a marked scavenging effect on superoxide, and also an effect on hydrogen peroxide, but not so much on 1,1-diphenyl-2-picrylhydrazyl radical. WSP had only a small amount of sulfhydryl groups. Thus, the sulfhydryl groups are not responsible for the scavenging activity of WSP.

Journal Article↗
PubMed1999

Cloning and expression of amino acid transporters from broad bean.

This work describes the isolation of a full-length (VfAAP2) and three partial amino acid transporter genes (VfAAPa, VfAAPb, VfAAPc) from broad bean (Vicia faba L.). The function of VfAAP2 was tested by heterologous expression in a yeast mutant deficient in proline uptake. VfAAP2 mediates proton-dependent proline uptake with an apparent Km of about 1 mM. Analysis of substrate specificity by competition experiments showed that aromatic amino acids, neutral aliphatic acids and L-citrulline are the best competitors, whereas basic amino acids do not compete with proline. Northern analysis indicates that all VfAAPs exhibit different patterns of expression. VfAAP2 is most strongly expressed in the stem and at a lower level in sink leaves and pods. VfAAPa, VfAAPb and VfAAPc are most strongly expressed in the flowers, but their expression in the other organs varies.

Amino Acid Sequence↗
PubMed1971

Characterization of cytoplasmic and chloroplast 5S ribosomal ribonucleic acid from broad-bean leaves.

Green leaves of the broad bean (Vicia faba) contain two 5S RNA components that can be separated from each other by polyacrylamide-gel electrophoresis. The major component is located in the larger subunit of cytoplasmic ribosomes, whereas the minor component occurs in the larger subunit of chloroplast ribosomes. Their electrophoretic mobilities relative to those of Escherichia coli 5S RNA (120 nucleotides) and plant 4S RNA (78 nucleotides) suggest that they consist of 118 and 122 nucleotide residues respectively. Thermal ;melting' profiles of plant cytoplasmic and chloroplast 5S RNA species at 260nm indicate the similarity of their secondary structures, not only to each other, but also to those of E. coli and mammalian 5S RNA species. The base compositions of the two plant 5S RNA species have more in common with each other than with the corresponding molecules from either E. coli or mammalian cells.

Acrylates↗
PubMed1971

The formation of glycoproteins in tissues of higher plants. Specific labelling with D-(1- 14 C)glucosamine.

1. Radioactivity from d-[l-(14)C]glucosamine is incorporated into ethanol-insoluble compounds of high molecular weight in a number of plant tissues, including roots of corn (Zea mays), callus cells of sycamore (Acer pseudoplatanus), axenic cultures of duckweed (Lemna minor) and germinating seedlings of corn, broad bean (Vicia faba) and barley (Hordeum vulgare). 2. Except in the case of Lemna, where some of the radioactivity was recovered in glucose, hydrolysis of these ethanol-insoluble materials with acid released [(14)C]glucosamine as the major radioactive product. 3. The labelled compounds isolated from Zea roots and the Acer cells are believed to be glycoproteins rather than polysaccharides on the basis of their solubility properties, their charge characteristics and their susceptibility to hydrolysis by 0.5m-potassium hydroxide and by the proteases trypsin and Pronase. Further, radioactive peptides were isolated and purified after Pronase treatment and shown to contain glucosamine as well as a number of amino acids. 4. The experiments therefore indicate that d-[(14)C]glucosamine can be used as a specific precursor of the amino sugar units of plant as well as animal glycoproteins.

Alkalies↗
PubMed1993

[Broad beans as a cause of acute hemolytic anemia].

A previously healthy 17-year-old Greek boy suddenly developed jaundice of sclerae and skin. In addition, physical examination revealed a pale appearance. He also reported feeling tired and weak. The haemoglobin level was 9.6 g/dl, lactate dehydrogenase activity 335 U/l, bilirubin concentration 3.2 mg/dl (direct bilirubin 0.7 mg/dl, indirect bilirubin 2.5 mg/dl), haptoglobin concentration 48.8 mg/dl. As haemolytic anaemia was assumed, direct questioning elicited the fact that the patient had, for the first time in his life, eaten 300 g of broad beans (Vicia faba) on each of two days, namely 3 and 2 days before the appearance of jaundice. Absence of glucose-6-phosphate dehydrogenase activity in the red blood corpuscles confirmed the diagnosis of favism. On symptomatic treatment both the enzyme activities and the bilirubin level fell to normal within one week, and the haemoglobin level was 15.7 g/dl after 4 weeks.

Acute Disease↗
PubMed2004

Amino acid requirement of growing pigs depending on amino acid efficiency and level of protein deposition 1st communication: lysine.

The study was conducted to evaluate the variability of efficiency of lysine utilisation in different feed proteins for growing pigs including wheat, unprocessed soybean flakes (SF), hydrothermal processed SF, corn gluten meal (CGM), two batches of soybean protein concentrate (SPC-1, SPC-2), different batches of peas (Pisum sativum) and field beans (Vicia faba). Data about efficiency of lysine are important for further conclusions related to lysine requirement in dependence on level of daily protein deposition. In N-balance studies 161 growing barrows (40-65 kg BW) of the genotype [Piétrain x (Duroc x Landrace)] were randomly allotted into 23 experimental treatments (n = 7) with diets in which lysine was the first limiting amino acid. Data from the N-balance trials were used to calculate efficiency of lysine and consequently lysine requirement based on an exponential N-utilisation model. Results from the present studies indicate that efficiency of lysine in different feed proteins varies in a very wide range and this variation greatly affected the calculated lysine requirement. Therefore, the variation in efficiency of lysine should be taken into account in requirement calculations and consequently in diet formulation for pigs. The results of model calculation for the lysine requirement depending on daily protein deposition (130, 145 and 160 g/d) and efficiency of lysine are given for different standards for comparison (g x kg(-0.67) x d(-1), g x d(-1) and percentage of lysine in the diet). The calculated lysine requirement of growing barrows (50 kg BW) corresponding to an average lysine efficiency was 15.5, 18.0 and 21.1 g/d for daily protein deposition of 130, 145 and 160 g, respectively. The results of the model calculation for 145 or 160 g daily protein deposition are in agreement with actual studies and recommendations of NRC (1998) and DLG (2002) for lysine supply.

Animal Feed↗
PubMed2004

Amino acid requirement of growing pigs depending on amino acid efficiency and level of protein deposition. 2nd communication: threonine.

This study was conducted to evaluate the variability of the efficiency of threonine in different feed proteins for growing pigs. This information is of importance for actual conclusions about threonine requirement within the exponential N-utilization model (Liebert and Gebhardt, 1986) used in our investigations. Wheat (as basal protein), high-protein soybean meal, low-protein soybean meal, rapeseed meal, field bean (Vicia faba), peas (Pisum sativum), corn gluten meal and soybean protein concentrate were used as protein sources. Fifty-six growing barrows (40-65 kg BW) of the genotype Piétrain x (Duroc x Landrace) were randomly allotted to eight N-balance experiments (n = 7). Diets were formulated with two main ingredients (wheat + one feed protein) with threonine as the first limiting amino acid in the mixture which was partly supplemented with crystalline amino acids. Based on N-balance data, the efficiency of threonine was determined in protein mixtures and individual feed proteins. Threonine requirement was calculated depending on efficiency of threonine and level of daily protein deposition. The results from the present studies indicate that the efficiency of threonine in different feed proteins varied in a wide range. Consequently, this factor has to be taken into account for requirement calculations. The threonine requirement depending on daily protein deposition (130, 145 and 160 g) and the efficiency of threonine according to different reference units (g/BW(kg)(-0.67)/d, g/d and % of threonine in the diet) were calculated. The threonine requirement of growing barrows (50 kg BW) corresponding to an average threonine efficiency was 8.52, 9.92 and 11.61 g/d for a daily protein deposition of 130, 145 and 160 g, respectively. The results for a daily protein deposition of 145 or 160 g are in agreement with actual studies and recommendations for threonine supply.

Animal Feed↗
PubMed1958

Structure and mode of formation of the nucleolus in meristematic cells of Vicia faba and Allium cepa.

Interphase nucleoli from Vicia faba and Allium cepa meristematic cells are roughly classified into two categories: (a) those that commonly show a rather homogeneous texture (except for small light spaces of various sizes) and frequently contain dense particles 140 A in diameter; (b) those found more frequently in Vicia characterized by a very sharp boundary between a dense outer cortex and a much lighter central core. The dense particles are not found in such nucleoli. In Allium the boundary is more irregular and dense particles are sometimes observed in the outer layer. Many nucleoli show a structure intermediate between these two types. They are characterized by a gradient of increasing density from the center to the periphery and occasionally contain dense 140 A granules. During interphase, certain nucleoli are closely associated with segments of chromatin strands which undoubtedly represent nucleolar organizing regions. The dense 140 A granules are followed during the mitotic cycle. In Allium, they are first seen in loose clusters between arms of late anaphase chromosomes where they become more concentrated in early telophase. The substance within which they are scattered slowly increases in density during that time until finally, the particles are limited to small bodies of distinctive character. Evidence is presented suggesting that these small prenucleolar bodies fuse during telophase to give rise to the mature interphase nucleoli. Similar events are described in Vicia material except that a coating of dense substance appears around telophase chromosomes before the formation of prenucleolar bodies.

Anaphase↗
PubMed1958

A cytological analysis of the antimetabolite activity of 5-hydroxyuracil in Vicia faba roots.

The effects of 5-hydroxyuracil (5-HU) (isobarbituric acid) upon cell elongation, mitosis, and DNA synthesis were studied in Vicia faba roots. 5-HU had no consistent effect upon root elongation. It blocked DNA synthesis (analyzed by photometric measurements of Feulgen dye in nuclei) during the first 6 hours of treatment; the block spontaneously disappeared by the 12th hour of treatment. Uracil and thymine had no effect upon this block of synthesis. Both thymidine and uridine reversed the block in 6 and 9 hours respectively. In all cases blockage of DNA synthesis was followed by inhibition of mitosis (determined by changes in the percentage of cells in mitosis) and resumption of DNA synthesis was followed by resumption of mitosis. Inhibition indices calculated from the mitotic data indicated a competitive relationship between 5-HU and thymidine and 5-HU and uridine. 5-HU is considered to block DNA synthesis by competing with thymidine for sites on enzymes involved in the synthesis. It is suggested that uridine reverses the block in synthesis by undergoing a conversion to thymidine.

Antimetabolites↗