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Sample records for barley peroxidase isozymes

  1. Barley peroxidase isozymes

    Science.gov (United States)

    Laugesen, Sabrina; Bak-Jensen, Kristian Sass; Hägglund, Per; Henriksen, Anette; Finnie, Christine; Svensson, Birte; Roepstorff, Peter

    2007-12-01

    Thirteen peroxidase spots on two-dimensional gels were identified by comprehensive proteome analysis of the barley seed. Mass spectrometry tracked multiple forms of three different peroxidase isozymes: barley seed peroxidase 1, barley seed-specific peroxidase BP1 and a not previously identified putative barley peroxidase. The presence of multiple spots for each of the isozymes reflected variations in post-translational glycosylation and protein truncation. Complete sequence coverage was achieved by using a series of proteases and chromatographic resins for sample preparation prior to mass spectrometric analysis. Distinct peroxidase spot patterns divided the 16 cultivars tested into two groups. The distribution of the three isozymes in different seed tissues (endosperm, embryo, and aleurone layer) suggested the peroxidases to play individual albeit partially overlapping roles during germination. In summary, a subset of three peroxidase isozymes was found to occur in the seed, whereas products of four other barley peroxidase genes were not detected. The present analysis documents the selective expression profiles and post-translational modifications of isozymes from a large plant gene family.

  2. Barley peroxidase isozymes. Expression and post-translational modification in mature seeds as identified by two-dimensional gel electrophoresis and mass spectrometry

    DEFF Research Database (Denmark)

    Laugesen, Sabrina; Bak-Jensen, Kristian Sass; Hägglund, Per

    2007-01-01

    putative barley peroxidase. The presence of multiple spots for each of the isozymes reflected variations in post-translational glycosylation and protein truncation. Complete sequence coverage was achieved by using a series of proteases and chromatographic resins for sample preparation prior to mass....... In summary, a subset of three peroxidase isozymes was found to occur in the seed, whereas products of four other barley peroxidase genes were not detected. The present analysis documents the selective expression profiles and post-translational modifications of isozymes from a large plant gene family. (C...

  3. Purification and some properties of peroxidase isozymes from pineapple stem.

    Science.gov (United States)

    Sung, H Y; Yu, R H; Chang, C T

    1993-01-01

    The enzyme peroxidase is widely distributed among the higher plants. Isozymes of peroxidase are known to occur in a variety of tissues in a large number of plant species. In this study, peroxidase isozymes were purified from the extract of pineapple stem through successive steps of ammonium sulfate fractionation, CM-Sepharose CL-6B chromatographies and DEAE-Sepharose CL-6B chromatographies. By these steps, twelve isozymes of peroxidase were obtained. Some properties of the isozymes were studied and compared.

  4. The Role of Peroxidase Isozymes in Resistance to Wheat Stem Rust Disease 1

    Science.gov (United States)

    Seevers, P. M.; Daly, J. M.; Catedral, F. F.

    1971-01-01

    In common with other disease situations, rust-resistant wheat leaves show a large increase in peroxidase activity during infection. Peroxidase isozymes from healthy or infected lines of wheat (Triticum aestivum L.) near isogenic for resistance and susceptibility to race 56 of Puccinia graminis tritici were separated by gel electrophoresis and the activity of each was estimated by photometric scanning. In order to ensure that the activity of isozymes observed on gels reflected the changes found in peroxidase enzymes assayed spectrophotometrically in extracts, a study was made of extraction procedures, substrates, and reaction conditions for both types of enzyme measurements. Of the 14 isozymes detected in both healthy and infected leaves, increases in only 1 (isozyme 9) were associated consistently with the development of resistant disease reaction at 20 C. Additional evidence was obtained to show that this isozyme can account for the increased peroxidase activity observed in extracts from resistant plants. When plants with high induced peroxidase activity due to resistance at 20 C were treated with ethylene or transferred to 25 C, they reverted to complete susceptibility. However, the disease-induced activity of isozyme 9 did not fall. The data suggest that, in this case, the association of peroxidase with resistance was a consequence of, not a determinant in, resistance. PMID:16657797

  5. The role of peroxidase isozymes in resistance to wheat stem rust disease.

    Science.gov (United States)

    Seevers, P M; Daly, J M; Catedral, F F

    1971-09-01

    In common with other disease situations, rust-resistant wheat leaves show a large increase in peroxidase activity during infection. Peroxidase isozymes from healthy or infected lines of wheat (Triticum aestivum L.) near isogenic for resistance and susceptibility to race 56 of Puccinia graminis tritici were separated by gel electrophoresis and the activity of each was estimated by photometric scanning. In order to ensure that the activity of isozymes observed on gels reflected the changes found in peroxidase enzymes assayed spectrophotometrically in extracts, a study was made of extraction procedures, substrates, and reaction conditions for both types of enzyme measurements. Of the 14 isozymes detected in both healthy and infected leaves, increases in only 1 (isozyme 9) were associated consistently with the development of resistant disease reaction at 20 C. Additional evidence was obtained to show that this isozyme can account for the increased peroxidase activity observed in extracts from resistant plants. When plants with high induced peroxidase activity due to resistance at 20 C were treated with ethylene or transferred to 25 C, they reverted to complete susceptibility. However, the disease-induced activity of isozyme 9 did not fall. The data suggest that, in this case, the association of peroxidase with resistance was a consequence of, not a determinant in, resistance.

  6. Comparative analysis of peroxidase profiles in Chinese kale (Brassica alboglabra L.): evaluation of leaf growth related isozymes.

    Science.gov (United States)

    Tang, Lei; Wang, Chenchen; Huang, Jiabao; Zhang, Jianhua; Mao, Zhonggui; Wang, Haiou

    2013-01-15

    Plant peroxidases (EC 1.11.1.7) with different isoforms catalyze various reactions in plant growth and development. However, it is difficult to elucidate the function of each isozyme in one plant. Here, we compared profiles of entire isozyme in young seedling and mature leaves of Chinese kale (Brassica alboglabra L.) on zymogram and ion exchange chromatography in order to investigate leaf growth related peroxidase isozymes. The results showed that four isozymes were constitutively expressed in kale leaves, whereas other two isozymes were induced in the mature leaves. The Mono Q ion exchange chromatography separated the six isozymes into two major groups due to the difference in their isoelectric points. The results suggested that although there were several isozymes in the leaves of Chinese kale, one isozyme functioned mainly through the leaf development. Two anionic isozymes with molecular weights lower than 32 kDa were considered mature related.

  7. Barley coleoptile peroxidases. Purification, molecular cloning, and induction by pathogens

    DEFF Research Database (Denmark)

    Kristensen, B.K.; Bloch, H.; Rasmussen, Søren Kjærsgård

    1999-01-01

    A cDNA clone encoding the Prx7 peroxidase from barley (Hordeum vulgare L.) predicted a 341-amino acid protein with a molecular weight of 36,515. N- and C-terminal putative signal peptides were present, suggesting a vacuolar location of the peroxidase. Immunoblotting and reverse-transcriptase poly...

  8. Kinetic and thermodynamic properties of two barley thioredoxin h isozymes, HvTrxh1 and HvTrxh2

    DEFF Research Database (Denmark)

    Maeda, Kenji; Hägglund, Per; Björnberg, Olof

    2010-01-01

    Barley thioredoxin h isozymes 1 (HvTrxh1) and barley thioredoxin h isozymes 2 (HvTrxh2) show distinct spatiotemporal distribution in germinating seeds. Using a novel approach involving measurement of bidirectional electron transfer rates between Escherichia coli thioredoxin, which exhibits redox......-dependent fluorescence, and the barley isozymes, reaction kinetics and thermodynamic properties were readily determined. The reaction constants were 60% higher for HvTrxh1 than HvTrxh2, while their redox potentials were very similar. The primary nucleophile, Cys(N), of the active site Trp-Cys(N)-Gly-Pro-Cys...

  9. Peroxidase profiling reveals genetic linkage between peroxidase gene clusters and basal host and non-host resistance to rusts and mildew in barley.

    Directory of Open Access Journals (Sweden)

    Ana M González

    Full Text Available BACKGROUND: Higher plants possess a large multigene family encoding secreted class III peroxidase (Prx proteins. Peroxidases appear to be associated with plant disease resistance based on observations of induction during disease challenge and the presence or absence of isozymes in resistant vs susceptible varieties. Despite these associations, there is no evidence that allelic variation of peroxidases directly determines levels of disease resistance. METHODOLOGY/PRINCIPAL FINDINGS: The current study introduces a new strategy called Prx-Profiling. We showed that with this strategy a large number of peroxidase genes can be mapped on the barley genome. In order to obtain an estimate of the total number of Prx clusters we followed a re-sampling procedure, which indicated that the barley genome contains about 40 peroxidase gene clusters. We examined the association between the Prxs mapped and the QTLs for resistance of barley to homologous and heterologous rusts, and to the barley powdery mildew fungus. We report that 61% of the QTLs for partial resistance to P. hordei, 61% of the QTLs for resistance to B. graminis and 47% of the QTLs for non-host resistance to other Puccinia species co-localize with Prx based markers. CONCLUSIONS/SIGNIFICANCE: We conclude that Prx-Profiling was effective in finding the genetic location of Prx genes on the barley genome. The finding that QTLs for basal resistance to rusts and powdery mildew fungi tend to co-locate with Prx clusters provides a base for exploring the functional role of Prx-related genes in determining natural differences in levels of basal resistance.

  10. Ethylene production and peroxidase activity in aphid-infested barley.

    Science.gov (United States)

    Argandoña, V H; Chaman, M; Cardemil, L; Muñoz, O; Zúñiga, G E; Corcuera, L J

    2001-01-01

    The purpose of this work was to investigate whether ethylene is involved in the oxidative and defensive responses of barley to the aphids Schizaphis graminum (biotype C) and Rhopalophum padi. The effect of aphid infestation on ethylene production was measured in two barley cultivars (Frontera and Aramir) that differ in their susceptibility to aphids. Ethylene evolution was higher in plants infested for 16 hr than in plants infested for 4 hr in both cultivars. Under aphid infestation, the production of ethylene was higher in cv. Frontera than in Aramir, the more aphid susceptible cultivar. Ethylene production also increases with the degree of infestation. Maximum ethylene evolution was detected after 16 hr when plants were infested with 10 or more aphids. Comparing the two species of aphids, Schizaphis graminum induced more ethylene evolution than Rhopalosiphum padi. Infestation with S. graminum increased hydrogen peroxide content and total soluble peroxidase activity in cv. Frontera, with a maximum level of H2O2 observed after 20 min of infestation and the maximum in soluble peroxidase activity after 30 min of infestation. When noninfested barley seedlings from cv. Frontera were exposed to ethylene, an increase in hydrogen peroxide and in total peroxidase activity was detected at levels similar to those of infested plants from cv. Frontera. When noninfested plants were treated with 40 ppm of ethylene, the maximum levels of H2O2 and soluble peroxidase activity were at 10 and 40 min, respectively. Ethylene also increased the activity of both cell-wall-bound peroxidases types (ionically and covalently bound), comparable with infestation. These results suggest that ethylene is involved in the oxidative responses of barley plants induced by infestation.

  11. The Quantum Mixed-Spin Heme State of Barley Peroxidase: A Paradigm for Class III Peroxidases

    Energy Technology Data Exchange (ETDEWEB)

    Howes, B.D.; Ma, J.; Marzocchi, M.P.; Schiodt, C.B.; Shelnutt, J.A.; Smulevich, G.; Welinder, K.G.; Zhang, J.

    1999-03-23

    Electronic absorption and resonance Raman (RR) spectra of the ferric form of barley grain peroxidase (BP 1) at various pH values both at room temperature and 20 K are . reported, together with EPR spectra at 10 K. The ferrous forms and the ferric complex with fluoride have also been studied. A quantum mechanically mixed-spin (QS) state has been identified. The QS heme species co-exists with 6- and 5-cHS heroes; the relative populations of these three spin states are found to be dependent on pH and temperature. However, the QS species remains in all cases the dominant heme spin species. Barley peroxidase appears to be further characterized by a splitting of the two vinyl stretching modes, indicating that the vinyl groups are differently conjugated with the porphyrin. An analysis of the presently available spectroscopic data for proteins from all three peroxidase classes suggests that the simultaneous occurrence of the QS heme state as well as the splitting of the two vinyl stretching modes is confined to class III enzymes. The former point is discussed in terms of the possible influences of heme deformations on heme spin state. It is found that moderate saddling alone is probably not enough to cause the QS state, although some saddling maybe necessary for the QS state.

  12. Effects of calcium ion concentration on starch hydrolysis of barley alpha-amylase isozymes.

    Science.gov (United States)

    Yuk, Jeong-Bin; Choi, Seung-Ho; Lee, Tae-Hee; Jang, Myoung-Uoon; Park, Jung-Mi; Yi, Ah-Rum; Svensson, Birte; Kim, Tae-Jip

    2008-04-01

    Barley alpha-amylase genes, amy1 and amy2, were separately cloned into the expression vector of pPICZalphaA and recombinant Pichia strains were established by homologous recombination. Both AMYs from Pichia shared almost identical hydrolysis patterns on short maltooligosaccharides to result in glucose, maltose, or maltotriose. Against insoluble blue starch, AMY1 showed the highest activity at 0.1-5 mM calcium concentration, whereas 15-20 mM was optimal for AMY2. On the hydrolysis of soluble starch, unexpectedly, there was no significant difference between AMYs with increase of calcium. However, the relative activity on various starch substrates was significantly different between AMYs, which supports that the isozymes are clearly distinguished from each other on the basis of their unique preferences for substrates.

  13. Effects of calcium ion concentration on starch hydrolysis of barley α-amylase isozymes

    DEFF Research Database (Denmark)

    Yuk, Jeong-Bin; Choi, Seung-Ho; Lee, Tae-Hee;

    2008-01-01

    Barley (x-amylase genes, amyl and amy2, were separately cloned into the expression vector of pPICZ alpha A and recombinant Pichia strains were established by homologous recombination. Both AMYs from Pichia shared almost identical hydrolysis patterns on short maltooligosaccharides to result...... in glucose, maltose, or maltotriose. Against insoluble blue starch, AMY1 showed the highest activity at 0.1-5 mM calcium concentration, whereas 15-20 mM was optimal for AMY2. On the hydrolysis of soluble starch, unexpectedly, there was no significant difference between AMYs with increase of calcium. However......, the relative activity on various starch substrates was significantly different between AMYs, which supports that the isozymes are clearly distinguished from each other on the basis of their unique preferences for substrates....

  14. Purification, characterization and stability of barley grain peroxidase BP1, a new type of plant peroxidase

    DEFF Research Database (Denmark)

    Rasmussen, Christine B; Henriksen, Anette; Abelskov, A. Katrine

    1997-01-01

    The major peroxidase of barley grain (BP 1) has enzymatic and spectroscopic properties that are very differeant from those of other known plant peroxidases (EC 1.11.1.7) and can therefore contribute to the understanding of the many physiological functions ascribed to these enzymes. To study...... the structure-function relationships of this unique model peroxidase, large-scale and laboratory-scale purifications have been developed. The two batches of pure BP 1 obtained were identical in their enzymatic and spectral properties, and confirmed that BP 1 is different from the prototypical horseradish...... peroxidase isoenzyme C (HRP C). However, when measuring the specific activity of BP 1 at pH 4.0 in the presence of 1 mM CaCl2, the enzyme was as competent as HRP C at neutral pH towards a variety of substrates (mM mg(-1) min(-1)): coniferyl alcohol (930+/-48), caffeic acid (795+/-53), ABTS (2,2(1)-azino...

  15. Expression of a defence-related intercellular barley peroxidase in transgenic tobacco

    DEFF Research Database (Denmark)

    Kristensen, B.K.; Brandt, J.; Bojsen, K.

    1997-01-01

    Tobacco plants (Nicotiana benthamiana L.) have been transformed with a T-DNA vector construct carrying the cDNA pBH6-301, encoding the major pathogen induced leaf peroxidase (Prx8) of barley, under control of an enhanced CaMV 35S promoter. Progeny from three independent transformants were analyzed...... genetically, phenotypically and biochemically. The T-DNA was steadily inherited through three generations. The barley peroxidase is expressed and sorted to the intercellular space in the transgenic tobacco plants. The peroxidase can be extracted from the intercellular space in two molecular forms from both...... barley and transgenic tobacco. The tobacco expressed forms are indistinguishable from the barley expressed forms as determined by analytical isoelectric focusing (pI 8.5) and Western-blotting. Staining for N-glycosylation showed that one form only was glycosylated. The N-terminus of purified Prx8 from...

  16. cDNA, amino acid carbohydrate sequence of barley seed-specific peroxidase BP 1

    DEFF Research Database (Denmark)

    Johansson, A.; Rasmussen, Søren Kjærsgård; Harthill, J.E.

    1992-01-01

    The major peroxidase of barley seed BP 1 was characterized. Previous studies showed a low carbohydrate content, low specific activity and tissue-specific expression, and suggested that this basic peroxidase could be particularly useful in the elucidation of the structure-function relationship...... plant modified-type structure, Man-alpha-1-6(Xyl-beta-1-2)Man-beta-1-4GlcNAc-beta-1-4(Fuc-alpha-1-3)GlcNAc. The BP 1 gene was RFLP-mapped on barley chromosome 3, and we propose Prx5 as the name for this new peroxidase locus....

  17. Peroxidase profiling reveals genetic linkage between peroxidase gene clusters and basal host and non-host resistance to rusts and mildew in barley

    NARCIS (Netherlands)

    Gonzalez, A.M.; Marcel, T.C.; Kohutova, Z.; Stam, P.; Linden, van der C.G.; Niks, R.E.

    2010-01-01

    Background - Higher plants possess a large multigene family encoding secreted class III peroxidase (Prx) proteins. Peroxidases appear to be associated with plant disease resistance based on observations of induction during disease challenge and the presence or absence of isozymes in resistant vs sus

  18. Involvement of peroxidase activity in developing somatic embryos of Medicago arborea L. Identification of an isozyme peroxidase as biochemical marker of somatic embryogenesis.

    Science.gov (United States)

    Gallego, Piedad; Martin, Luisa; Blazquez, Antonio; Guerra, Hilario; Villalobos, Nieves

    2014-01-15

    The legume Medicago arborea L. is very interesting as regards the regeneration of marginal arid soils. The problem is that it does not have a good germinative yield. It was therefore decided to regenerate via somatic embryogenesis and find a marker of embryogenic potential. In this study, peroxidase activity was evaluated in non-embryogenic and embryogenic calli from M. arborea L. A decrease in soluble peroxidase activity is observed in its embryonic calli at the time at which the somatic embryos begin to appear. This activity is always lower in embryonic calli than in non-embryonic ones (unlike what happens in the case of wall-bound peroxidases). These results suggest that peroxidases can be considered to be enzymes involved in somatic embryogenesis in M. arborea. In addition, isozyme analyses were carried out on protein extracts using polyacrylamide gel electrophoresis. The band called P5 was detected only in embryogenic cultures at very early stages of development. This band was digested with trypsin and analyzed using linear ion trap (LTQ) mass spectrometer. In P5 isoform a peroxidase-L-ascorbate peroxidase was identified. It can be used as a marker that allows the identification of embryological potential.

  19. CDNA cloning, characterization and expression of an endosperm-specific barley peroxidase

    DEFF Research Database (Denmark)

    Rasmussen, Søren Kjærsgård; Welinder, K.G.; Hejgaard, J.

    1991-01-01

    A barley peroxidase (BP 1) of pI ca. 8.5 and M(r) 37000 has been purified from mature barley grains. Using antibodies towards peroxidase BP 1, a cDNA clone (pcR7) was isolated from cDNA expression library. The nucleotide sequence of pcR7 gave a derived amino acid sequence identical to the 158 C......-terminal amino acid residues of mature BP 1. The clone pcR7 encodes an additional C-terminal sequence of 22 residues, which apparently are removed during processing. BP 1 is less than 50% identical to other sequenced plant peroxidases. Analyses of RNA and protein from aleurone, endosperm and embryo tissue showed...

  20. Induction of soluble and cell wall peroxidases by aphid infestation in barley.

    Science.gov (United States)

    Chaman, M E; Corcuera, L J; Zúñiga, G E; Cardemil, L; Argandoña, V H

    2001-05-01

    Peroxidase enzymes have been found in soluble, ionically bound, and covalently bound forms and have been implicated in several physiological processes in plants. This paper investigates the effect of aphid infestation on soluble and bound-cell wall peroxidase activity and bound-cell wall isoform changes of barley plants. Peroxidase activity was measured in control plants and plants infested with the aphid Schizaphis graminum (Rondani). The activity of soluble peroxidases increased with time of infestation, older plants being more affected than younger ones. The increase in bound-cell wall peroxidase activity as a function of age was higher in infested than in control plants, being higher in ionically bound than in covalently bound peroxidases. When the aphids were removed from plants, the activities of both types of peroxidases decreased to control levels. Isoelectrofocusing analyses of the ionically bound peroxidases showed changes in the isoform pattern. A new isoform was induced by infestation. The activities of all covalently bound isoforms increased after infestation. The physiological implications of these changes are discussed.

  1. Interactions of barley alpha-amylase isozymes with Ca2+, substrates and proteinaceous inhibitors

    DEFF Research Database (Denmark)

    Abou Hachem, Maher; Bozonnet, Sophie; Willemoes, Martin

    2006-01-01

    discovered 'sugar tongs' site in domain C of AMY1 is thus critical for binding to starch granules. Furthermore, mutations of binding sites mostly reduced the degree of multiple attack in amylose hydrolysis. AMY1 has higher substrate affinity than AMY2, but isozyme chimeras with AMY2 domain C and other...

  2. Spatio-temporal profiling and degradation of α-amylase isozymes during barley seed germination

    DEFF Research Database (Denmark)

    Bak-Jensen, K.S.; Laugesen, S.; Østergaard, O.

    2007-01-01

    Ten genes from two multigene families encode barley alpha-amylases. To gain insight into the occurrence and fate of individual isoforms during seed germination, the alpha-amylase repertoire was mapped by using a proteomics approach consisting of 2D gel electrophoresis, western blotting, and mass...

  3. Spatio-temporal profiling and degradation of alpha-amylase isozymes during barley seed germination

    DEFF Research Database (Denmark)

    Bak-Jensen, K.S.; Laugesen, Sabrina; Østergaard, Ole

    2007-01-01

    Ten genes from two multigene families encode barley alpha-amylases. To gain insight into the occurrence and fate of individual isoforms during seed germination, the alpha-amylase repertoire was mapped by using a proteomics approach consisting of 2D gel electrophoresis, western blotting, and mass...... increased during germination. Assessing the fragment minimum chain length by peptide mass fingerprinting suggested that alpha-amylase 2 ( gi vertical bar 4699831) initially was cleaved just prior to domain B that protrudes from the (beta alpha)(8)-barrel between beta-strand 3 and alpha-helix 3, followed...... by cleavage on the C-terminal side of domain B and near the C-terminus. Only two shorter fragments were identified of the other alpha-amylase 2 (gi vertical bar 166985). The 2D gels of dissected tissues showed alpha-amylase degradation to be confined to endosperm. In contrast, the aleurone layer contained...

  4. A redox-dependent dimerization switch regulates activity and tolerance for reactive oxygen species of barley seed glutathione peroxidase

    DEFF Research Database (Denmark)

    Navrot, Nicolas; Skjoldager, Nicklas; Bunkenborg, Jakob

    2015-01-01

    Monomeric and dimeric forms of recombinant barley (Hordeum vulgare subsp. vulgare) glutathione peroxidase 2 (HvGpx2) are demonstrated to display distinctly different functional properties in vitro. Monomeric HvGpx2 thus has five fold higher catalytic efficiency than the dimer towards tert-butyl h...... active, but more oxidation-resistant dimer. ...

  5. Barley seed proteomics from spots to structures

    DEFF Research Database (Denmark)

    Finnie, Christine; Svensson, Birte

    2009-01-01

    with information from rice and other cereals facilitate identification of barley proteins. Several hundred barley seed proteins are identified and lower abundance proteins including membrane proteins are now being analysed. In the present review we focus on variation in protein profiles of seed tissues during...... forms on 2D-gels. Specific protein families, including peroxidases and alpha-amylases have been subjected to in-depth analysis resulting in characterisation of different isozymes, post-translational. modifications and processing. A functional proteomics study focusing on the seed thioredoxin system has...

  6. Assessment of peroxidase isozyme marker-based model for cross identifications in hybrids (F(1)) of urdbean [ Vigna mungo (L.) Hepper].

    Science.gov (United States)

    Upadhyay, R.; Shukla, A.; Gaur, K.

    2002-12-01

    Four hybrids (4 F(1)s) were chosen out of crosses in the urdbean [ Vigna mungo (L.) Hepper, 2n = 22] having contrasting morphological characters. Zymograms for isozyme peroxidase were drawn from the patterns obtained from parents and their respective F(1) hybrids on the basis of relative similarities to parental bands. The selfed or crossed nature of hybrid pods was determined from the zymograms and their analysis. The number of bands and their intensities gave an idea about the extent of crossing in F(1) populations. Genetic identity (I) values were indicative of their selfed nature. Dendrograms were constructed on the basis of genetic identity values to display the relative similarities between the populations. Analysis was based on individual pods to confirm their hybrid or selfed nature. Possible use of this technique for identification of F(1) pods and elimination of selfed pods might be implemented to shorten the breeding operations during crossing.

  7. Mechanisms of Salt Tolerance in Transgenic Arabidopsis thaliana Carrying a Peroxisomal Ascorbate Peroxidase Gene from Barley

    Institute of Scientific and Technical Information of China (English)

    XU Wei-Feng; SHI Wei-Ming; A. UEDA; T. TAKABE

    2008-01-01

    Ascorbate peroxidases (APX), localized in the cytosol, peroxisome, mitochondria, and chloroplasts of plant cells,catalyze the reduction of H2O2 to water by using ascorbic acid as the specific electron donor. To determine the role of peroxisomal type ascorbate peroxidasc (pAPX), an antioxidant enzyme, in protection against salt-induced oxidative stress, transgenic Arabidopsis thaliana plant carrying a pAPX gene (HvAPX1) from barley (Hordeum vulgare L.) was analyzed. The transgenic line pAPX3 was found to be more tolerant to salt stress than the wild type. Irrespective of salt stress, there were no significant differences in Na+, K+, Ca2+, and Mg2+ contents and the ratio of K+ to Na+ between pAPX3 and the wild type. Clearly, the salt tolerance in pAPX3 was not due to the maintenance and reestablishment of cellular ion homeostasis. However, the degree of H2O2 and lipid peroxidation (measured as the levels of malondialdehyde)accumulation under salt stress was higher in the wild type than in pAPX3. The mechanism of salt tolerance in transgenic pAPX3 can thus be explained by reduction of oxidative stress injury. Under all conditions tested, activities of superoxide,glutathionc reductase, and catalase were not significantly different between pAPX3 and the wild type. In contrast, the activity of APX was significantly higher in the transgcnic plant than in wild type under salt stress. These results suggested that in higher plants, HvAPX1 played an important role in salt tolerance and was a candidate gene for developing salt-tolerant crop plants.

  8. Genetical control and linkage relationships of isozyme markers in sugar beet (B. vulgaris L.) : 1. Isocitrate dehydrogenase, adenylate kinase, phosphoglucomutase, glucose phosphate isomerase and cathodal peroxidase.

    Science.gov (United States)

    Smed, E; Van Geyt, J P; Oleo, M

    1989-07-01

    Five isozyme systems were genetically investigated. The different separation techniques, the developmental expression and the use as marker system in sugar beet genetics and breeding is discussed. Isocitrate dehydrogenase was controlled by two genes. The gene products form inter- as well as intralocus dimers, even with the gene products of the Icd gene in B. procumbens and B. patellaris. Adenylate kinase was controlled by one gene. Three different allelic forms were detected, which were active as monomeric proteins. Glucose phosphate isomerase showed two zones of activity. One zone was polymorphic. Three allelic variants, active as dimers, were found. Phosphoglucomutase also showed two major zones of activity. One zone was polymorphic and coded for monomeric enzymes. Two allelic forms were found in the accessions studied. The cathodal peroxidase system was controlled by two independent genes, of which only one was polymorphic. The gene products are active as monomers. Linkage was found between red hypocotyl color (R) and Icd 2. Pgm 1, Gpi 2, Ak 1 and the Icd 2-R linkage group segregated independently.

  9. Novel promoter sequence required for manganese regulation of manganese peroxidase isozyme 1 gene expression in Phanerochaete chrysosporium.

    Science.gov (United States)

    Ma, Biao; Mayfield, Mary B; Godfrey, Bruce J; Gold, Michael H

    2004-06-01

    Manganese peroxidase (MnP) is a major, extracellular component of the lignin-degrading system produced by the wood-rotting basidiomycetous fungus Phanerochaete chrysosporium. The transcription of MnP-encoding genes (mnps) in P. chrysosporium occurs as a secondary metabolic event, triggered by nutrient-nitrogen limitation. In addition, mnp expression occurs only under Mn2+ supplementation. Using a reporter system based on the enhanced green fluorescent protein gene (egfp), we have characterized the P. chrysosporium mnp1 promoter by examining the effects of deletion, replacement, and translocation mutations on mnp1 promoter-directed egfp expression. The 1,528-bp mnp1 promoter fragment drives egfp expression only under Mn2+-sufficient, nitrogen-limiting conditions, as required for endogenous MnP production. However, deletion of a 48-bp fragment, residing 521 bp upstream of the translation start codon in the mnp1 promoter, or replacement of this fragment with an unrelated sequence resulted in egfp expression under nitrogen limitation, both in the absence and presence of exogenous Mn2+. Translocation of the 48-bp fragment to a site 120 bp downstream of its original location resulted in Mn2+-dependent egfp expression under conditions similar to those observed with the wild-type mnp1 promoter. These results suggest that the 48-bp fragment contains at least one Mn2+-responsive cis element. Additional promoter-deletion experiments suggested that the Mn2+ element(s) is located within the 33-bp sequence at the 3' end of the 48-bp fragment. This is the first promoter sequence containing a Mn2+-responsive element(s) to be characterized in any eukaryotic organism. Copyright 2004 American Society for Microbiology

  10. Aluminum-induced cell death of barley-root border cells is correlated with peroxidase- and oxalate oxidase-mediated hydrogen peroxide production.

    Science.gov (United States)

    Tamás, L; Budíková, S; Huttová, J; Mistrík, I; Simonovicová, M; Siroká, B

    2005-06-01

    The function of root border cells (RBC) during aluminum (Al) stress and the involvement of oxalate oxidase, peroxidase and H(2)O(2) generation in Al toxicity were studied in barley roots. Our results suggest that RBC effectively protect the barley root tip from Al relative to the situation in roots cultivated in hydroponics where RBC are not sustained in the area surrounding the root tip. The removal of RBC from Al-treated roots increased root growth inhibition, Al and Evans blue uptake, inhibition of RBC production, the level of dead RBC, peroxidase and oxalate oxidase activity and the production of H(2)O(2). Our results suggest that even though RBC actively produce active oxygen species during Al stress, their role in the protection of root tips against Al toxicity is to chelate Al in their dead cell body.

  11. Insights into the "pair of sugar tongs" surface binding site in barley alpha-amylase isozymes and crystallization of appropriate sugar tongs mutants

    DEFF Research Database (Denmark)

    Tranier, S.; Deville, K.; Robert, X.

    2005-01-01

    Recently, the three-dimensional structure of AMY1 in complex with a thio-maltotetraose (thio-DP4) has contributed to the understanding of the isozyme differences between AMY1 and AMY2, particularly the higher activity of AMY1 on starch granules. Indeed, this structure reveals the presence of an a...

  12. Insights into the "pair of sugar tongs" surface binding site in barley alpha-amylase isozymes and crystallization of appropriate sugar tongs mutants

    DEFF Research Database (Denmark)

    Tranier, S.; Deville, K.; Robert, X.

    2005-01-01

    Recently, the three-dimensional structure of AMY1 in complex with a thio-maltotetraose (thio-DP4) has contributed to the understanding of the isozyme differences between AMY1 and AMY2, particularly the higher activity of AMY1 on starch granules. Indeed, this structure reveals the presence of an a...

  13. COMPARED ANALYSIS OF CATALASE AND PEROXIDASE ACTIVITY IN CELLULOLYTIC FUNGUS TRICHODERMA REESEI GROWN ON MEDIUM WITH DIFFERENT CONCENTRATIONS OF GRINDED WHEAT AND BARLEY STRAWS

    Directory of Open Access Journals (Sweden)

    Mihaela Cristica

    2010-09-01

    Full Text Available The purpose of this study was to assess the evolution of catalase and peroxidase activity in Trichoderma reesei grown on medium containing grinded wheat and barley straws. Carbon source of cultivation medium - glucose was replaced by various concentrations of grinded wheat and barley straws, finally resulting three experimental variants as follows: V1 = 20 g/l, V2 = 30 g/l, V3 = 40 g/l. ĂŽn addition to these variants a control sample was added in which composition remainded unchanged. The catalase activity was determined by spectrophotometric Sinha method (Artenie et al., 2008 while peroxidase activity was assesed using the o-dianisidine method (Cojocaru, 2009. Enzymatic determinations were carried out at 7 and 14 days from inoculation, in both fungus mycelium and culture liquid. The enzymatic assay showed significant differences between determinations intervals and work variants. Enzyme activity is influenced by the age of fungus and by the different nature of the substrate used.

  14. Oligosaccharide binding to barley alpha-amylase 1

    DEFF Research Database (Denmark)

    Robert, X.; Haser, R.; Mori, H.

    2005-01-01

    Enzymatic subsite mapping earlier predicted 10 binding subsites in the active site substrate binding cleft of barley alpha-amylase isozymes. The three-dimensional structures of the oligosaccharide complexes with barley alpha-amylase isozyme 1 (AMY1) described here give for the first time a thorough...... in barley alpha-amylase isozyme 2 (AMY2), and the sugar binding modes are compared between the two isozymes. The "sugar tongs" surface binding site discovered in the AMY1-thio-DP4 complex is confirmed in the present work. A site that putatively serves as an entrance for the substrate to the active site...

  15. Isozyme Analysis on Different Varieties of Sugarcane

    Directory of Open Access Journals (Sweden)

    Johnson M.

    2012-05-01

    Full Text Available Isozymic and protein diversity among five sugarcane varieties viz., Co 6304, Co 85019, Co 8371, Co 89003 and Co 91010 were studied to understand the varietal interrelationship and to identify the biochemical marker for the disease resistance and stress tolerance. The standard technique of vertical gel electrophoresis PAGE was employed for size separation of isozymes. The gel was stained with different staining solutions for different isozyme systems viz. peroxidase, esterase, acid phosphatase, alkaline phosphatase and proteins. Rf values of the banding profiles, similarity index and variation between the varieties were analysed. Among the four enzyme systems, peroxidase profile reveals the difference between the disease resistant / susceptible and abiotic stress tolerant / non tolerant varieties. The two isoperoxidase bands with Rf values 0.62 and 0.66 showed their presence in disease resistant and abiotic tolerant varieties. The presence of two marker bands (0.62, 0.66 of resistant and stress tolerant varieties suggest that the variety Co 6304 may also be resistant to smut, wilt and moderately resistant to red rot and tolerant to drought.

  16. Oligosaccharide binding to barley alpha-amylase 1

    DEFF Research Database (Denmark)

    Robert, X.; Haser, R.; Mori, H.;

    2005-01-01

    Enzymatic subsite mapping earlier predicted 10 binding subsites in the active site substrate binding cleft of barley alpha-amylase isozymes. The three-dimensional structures of the oligosaccharide complexes with barley alpha-amylase isozyme 1 (AMY1) described here give for the first time a thorough...... insight into the substrate binding by describing residues defining 9 subsites, namely -7 through +2. These structures support that the pseudotetrasaccharide inhibitor acarbose is hydrolyzed by the active enzymes. Moreover, sugar binding was observed to the starch granule-binding site previously determined...... in barley alpha-amylase isozyme 2 (AMY2), and the sugar binding modes are compared between the two isozymes. The "sugar tongs" surface binding site discovered in the AMY1-thio-DP4 complex is confirmed in the present work. A site that putatively serves as an entrance for the substrate to the active site...

  17. Characterisation of taro (Colocasia esculenta based on morphological and isozymic patterns markers

    Directory of Open Access Journals (Sweden)

    SUGIYARTO

    2011-03-01

    Full Text Available Trimanto, Sajidan, Sugiyarto. 2011. Characterization of taro (Colocasia esculenta based on morphological and isozymic patterns markers. Nusantara Bioscience: 7-14. The aims of this research were to find out: (i the variety of Colocasia esculenta based on the morphological characteristics; (ii the variety of C. esculenta based on the isozymic banding pattern; and (iii the correlation of genetic distance based on the morphological characteristics and isozymic banding pattern. Survey research conducted in the Karanganyar district, which include high, medium and low altitude. The sample was taken using random purposive sampling technique, including 9 sampling points. The morphological data was elaborated descriptively and then made dendogram. The data on isozymic banding pattern was analyzed quantitatively based on the presence or absence of bands appeared on the gel, and then made dendogram. The correlation based on the morphological characteristics and isozymic banding pattern were analyzed based on the product-moment correlation coefficient with goodness of fit criterion. The result showed : (i in Karanganyar was founded 10 variety of C. esculenta; (ii morphological characteristics are not affected by altitude; (iii isozymic banding pattern of peroxides forms 14 banding patterns, esterase forms 11 banding patterns and shikimic dehydrogenase forms 15 banding patterns; (iv the correlation of morphological data and the isozymic banding pattern of peroxidase has good correlation (0.893542288 while esterase and shikimic dehydrogenase isozymes have very good correlation (0.917557716 and 0.9121985446; (v isozymic banding pattern of data supports the morphological character data.

  18. 不同产地、不同年限人参中3种同工酶活力比较%Comparison of the activities of peroxidase, catalase, malate dehydrogenase isozymes in Radix Ginseng from different sources

    Institute of Scientific and Technical Information of China (English)

    刘宏; 赵雨; 邢楠楠; 张惠; 李红艳

    2011-01-01

    目的:对不同产地,不同生长年限(4年、5年)的人参中过氧化物酶(Peroxidase POD)、过氧化氢酶(Catalase CAT)、苹果酸脱氢酶(Malate Dehydrogenase MDH)活力进行比较.方法:采用中性缓冲液提取总蛋白,应用愈创木酚比色法测定过氧化物酶活力,过氧化氢比色法测定过氧化氢酶活力,草酰乙酸比色法测定苹果酸脱氢酶活力.结果:不同产地人参的同工酶活力存在一定差异,4年、5年生人参同工酶活力具有相似性,差异较小.结论:POD、CAT、MDH的活力可以作为人参品种鉴定及药材优选的评价指标.

  19. Molecular size and net charge of pathogenesis-related enzymes from barley (Hordeum vulgare L., v. Karat) infected with Drechslera teres f. teres (Sacch.) Shoem.

    Science.gov (United States)

    Rothe, G M; Welschbillig, N; Reiss, E

    1998-05-01

    Molecular size and net charge of isoforms of pathogenesis-related (PR) chitinase, beta-1,3-glucanase and peroxidase were studied in uninfected barley (Hordeum vulgare L., v. Karat) leaves and in barley leaves infected with the pathogenic fungus Drechslera teres f. teres (Sacch.) Shoem. Molecular characteristics were determined by time-dependent polyacrylamide gradient gel electrophoresis under native conditions and by applying an extended version of the computer program MOL-MASS (Rothe, G. M., Weidmann, H., Electrophoresis 1991, 12, 703-709). Uninfected barley leaves contained predominantly one peroxidase isozyme but also three very weak peroxidases. Activities of all of these three peroxidases increased considerably after infection with Drechslera teres. The molecular masses of peroxidases 1 and 3 were estimated to be 38 +/- 5 and 42 +/- 7 kDa and their apparent valences at pH 8.4 were Z = 3.13 and 3.20, respectively. Amongst the chitinase isoforms, chitinase 1 and chitinase 2 appeared after infection, while chitinase 3 was also observed in uninfected leaves of barley. The molecular mass of chitinase 3 (31 +/- 6 kDa; f/fo = 1.20) was larger than that of chitinase 1 (20 +/- 2 kDa; f/fo = 1.04) and chitinase 2 (23 +/- 3 kDa; f/fo = 1.06). The valence of constitutive chitinase 3 (Z = 1.44 +/- 0.81) at pH 8.4 was lower than that of adaptive chitinase 1 (Z = 3.27 +/- 1.02) and chitinase 2 (Z = 2.96 +/- 1.38). Infection of barley leaves with Drechslera teres also induced the hydrolytic enzyme beta-1,3-glucanase 1; beta-1,3-glucanase 2 appeared in uninfected and in infected leaves. Constitutive beta-1,3-glucanase 2 was smaller (molecular mass 19 +/- kDa; f/fo = 1.05) than adaptive beta-1,3-glucanase 1 (molecular mass 26 +/- 4 kDa; f/fo = 1.07). The valence of adaptive beta-1,3-glucanase 1 (Z = 9.58 +/- 4.17) was approximately threefold that of beta-1,3-glucanase 2 (Z = 2.80 +/- 0.93).

  20. Reconstitution of cyanogenesis in barley (Hordeum vulgare L.) and its implications for resistance against the barley powdery mildew fungus.

    Science.gov (United States)

    Nielsen, Kirsten A; Hrmova, Maria; Nielsen, Janni Nyvang; Forslund, Karin; Ebert, Stefan; Olsen, Carl E; Fincher, Geoffrey B; Møller, Birger Lindberg

    2006-04-01

    Barley (Hordeum vulgare L.) produces a leucine-derived cyanogenic beta-D-glucoside, epiheterodendrin that accumulates specifically in leaf epidermis. Barley leaves are not cyanogenic, i.e. they do not possess the ability to release hydrogen cyanide, because they lack a cyanide releasing beta-D-glucosidase. Cyanogenesis was reconstituted in barley leaf epidermal cells through single cell expression of a cDNA encoding dhurrinase-2, a cyanogenic beta-D-glucosidase from sorghum. This resulted in a 35-60% reduction in colonization rate by an obligate parasite Blumeria graminis f. sp. hordei, the causal agent of barley powdery mildew. A database search for barley homologues of dhurrinase-2 identified a (1,4)-beta-D-glucan exohydrolase isozyme betaII that is located in the starchy endosperm of barley grain. The purified barley (1,4)-beta-D-glucan exohydrolase isozyme betaII was found to hydrolyze the cyanogenic beta-D-glucosides, epiheterodendrin and dhurrin. Molecular modelling of its active site based on the crystal structure of linamarase from white clover, demonstrated that the disposition of the catalytic active amino acid residues was structurally conserved. Epiheterodendrin stimulated appressoria and appressorial hook formation of B. graminis in vitro, suggesting that loss of cyanogenesis in barley leaves has enabled the fungus to utilize the presence of epiheterodendrin to facilitate host recognition and to establish infection.

  1. Barley germination

    DEFF Research Database (Denmark)

    Daneri-Castro, Sergio N.; Svensson, Birte; Roberts, Thomas H.

    2016-01-01

    conditions continue to be key to discovering the roles of individual protein forms and posttranslational modifications, such as glycosylation. Activity-based proteomics, particularly in combination with new gene editing technologies, has great potential to elucidate the network of enzymes in barley...

  2. Characterization of lignin and Mn peroxidases from Phanerochaete chrysosporium

    Energy Technology Data Exchange (ETDEWEB)

    1991-01-01

    Long-term objectives are to elucidate the role and mechanism of the various isozymes in lignin biodegradation. Work is described on electrochemical studies on lignin and Mn peroxidases. This study was performed to investigate the structural aspects which confer the lignin and Mn peroxidases with their high reactivity. The experimentally determined redox potential of the Fe{sup 3+}/Fe{sup 2+} couple for the lignin peroxidase isozymes H1, H2, H8 and H10 are very similar, near-130 mV. The redox potential for the Mn peroxidase isozymes H3 and H4 are similar to each other ({minus}88 mV and {minus}95 mV, respectively) and are more positive than the lignin peroxidases. The higher redox potential for the Fe{sup 3+}/Fe{sup 2+} couple is consistent with the heme active site of these fungal peroxidases being more electron deficient. To investigate the accessibility of the heme active site to the substrate which is oxidized (veratryl alcohol and Mn (II)), we investigated whether these substrates had any affect on the redox potential of the heme. The E{sub m7} value for lignin and Mn peroxidases are not affected by their respective substrates, veratryl alcohol and Mn (II). These results suggest that substrates do not directly interact with the ferric heme-iron as axial ligands. This is consistent with the present model for peroxidase catalysis. Suicide inhibitor (1) and nmr studies (2) indicate that the heme-iron of horseradish peroxidase (HRP) is not fully accessible to bulky substrates occur at the periphery of the heme.

  3. Short Communication: Variation in isozymic pattern of germplasm from three ginger (Zingiber officinale varieties

    Directory of Open Access Journals (Sweden)

    AHMAD DWI SETYAWAN

    2014-06-01

    Full Text Available Setyawan AD, Wiryanto, Suranto, Bermawie N. 2014. Variation in isozymic pattern of germplasm from three of ginger (Zingiber officinale varieties. Nusantara Bioscience 6: 86-93. Ginger (Zingiber officinale Rosc. has long been as spices, flavoring agent and raw material for herbal medicines. In Indonesia, there were three varieties based on color and size of the rhizome, i.e. gajah (big-white ginger, merah (red ginger, and emprit (small-white ginger. This research was conducted to find out: (i isozymic pattern of three ginger varieties, and (ii phylogenetic relationship of those three varieties. The plant materials were gathered from Wonogiri, Surakarta and Kulonprogo, Yogyakarta. Two enzyme systems, namely esterase (EST and peroxidase (PER, PRX were used in this study. The relationship among ginger varieties was determined by UPGMA. The result indicated that EST showed two bands (i.e. Rf 0.04 and 0.10, and PRX showed six bands (i.e. Rf 0.04, 0.05, 0.09, 0.10, 0.11, and 0.15. Peroxidase produce more numerous and more diverse isozymic bands than esterase, resulting in a more complex relationship. The data used to compile dendrogram affect the grouping; the more data used, the more obvious clustering of accessions in a population. Dendrogam generated from esterase and peroxidase banding patterns produced distinct clusters based on varieties and location.

  4. Characterization of white grubs (Melolonthidae: Coleoptera at salak pondoh agroecosystem in Mount Merapi based on isozymic banding patterns

    Directory of Open Access Journals (Sweden)

    SRI WARDANI

    2009-03-01

    Full Text Available Wardani S, Sugiyarto. 2009. Characterization of white grubs (Melolonthidae: Coleoptera at salak pondoh agroecosystem in Mount Merapi based on isozymic banding patterns. Nusantara Bioscience 1: 38-42. The aim of this research is to know the characteristics of white grubs (Melolonthidae: Coleoptera based on isozyme banding patterns. This research was conducted at Sleman, Yogyakarta and Magelang-Central Java for the morphological purposes. The sample was taken from 5 places with different height in wich 5 samples were taken from each location. The method used in this research was polyacrylamide gel electrophoresis (PAGE using the vertical type. The enzyme system used in this research were peroxidase and esterase to detect the isozyme banding patterns. The results showed that there was a variation in isozyme banding patterns of white grubs (Melolonthidae: Coleoptera at salak pondoh agroecosystem in Mount Merapi’s slope (peroxidase in station II and IV while esterase in station III and V. It’s mean that genetic variation on white grubs population at salak pondoh agroecosystem in Mount Merapi’s slope was found. The environmental condition also contributed to the influence of the appear of isozyme banding pattern’s variation because each location had a different condition.

  5. Overexpression, purification, and characterization of recombinant barley alpha-amylases 1 and 2 secreted by the methylotrophic yeast Pichia pastoris

    DEFF Research Database (Denmark)

    Juge, N; Andersen, Jens S.; Tull, D

    1996-01-01

    Recombinant barley alpha-amylase isozymes 1 and 2 were secreted by Pichia pastoris at up to 50 and 1 mg/liter, respectively, representing approximately a 50-fold increase compared to the levels of the heterologous expression by Saccharomyces cerevisiae. The cDNA clones E or pM/C encoding isozymes 1...... on beta-cyclodextrin-Sepharose. The N-terminal sequence, pI, and Mr indicated that native-like processing took place. Electrospray ionization mass spectrometry, however, revealed microheterogeneity for recombinant isozyme 1. While Mr of one recombinant isozyme 1 form of 45,452 was in excellent agreement...... with a value of 45,447 calculated from the sequence, liquid chromatography/mass spectrometry of endo Lys C-generated peptides followed by tandem mass spectrometry on a nanoelectrospray ionization/mass spectrometry/mass spectrometry system identified additional recombinant isozyme 1 forms to be glycosylated...

  6. Brewing with fractionated barley

    OpenAIRE

    Donkelaar, van, CC René

    2016-01-01

    Brewing with fractionated barley Beer is a globally consumed beverage, which is produced from malted barley, water, hops and yeast. In recent years, the use of unmalted barley and exogenous enzymes have become more popular because they enable simpler processing and reduced environmental impact. Raw barley, however, contains less endogenous enzymes and more undesired components for the use of beer brewing, compared to malted barley.  The overall aim of this thesis is to investigate how ba...

  7. Short Communication: Comparisons of isozyme diversity in local Java cardamom (Amomum compactum and true cardamom (Elettaria cardamomum

    Directory of Open Access Journals (Sweden)

    AHMAD DWI SETYAWAN

    2014-05-01

    Full Text Available Setyawan AD, Wiryanto, Suranto, Bermawie N, Sudarmono. 2014. Comparisons of isozyme diversity in local Java cardamom (Amomum compactum and true cardamom (Elettaria cardamomum. Nusantara Bioscience 6: 94-101. Fruits of Java cardamoms (Amomum compactum and true cardamoms (Elettaria cardamomum had long been used as spices, flavoring agent, garnishing plants etc. This research was conducted to find out: (i variation of isozymic bands in some population of Java cardamoms and true cardamoms; and (ii phylogenetic relationship of these cardamoms based on variation of isozymic bands. Plant material (i.e. rhizome of Java cardamoms was collected from Bogor Botanical Garden, and plant material of true cardamoms was gathered from Indonesian Medicinal and Aromatic Crops Research Institute, Bogor, Indonesia. Ten accessions were assayed in every population. The two isozymic systems was assayed, namely esterase (EST and peroxidase (PER, PRX. Phylogenetic relationship was determined by UPGMA method. The results showed that esterase gave nine isozymic bands, i.e. Rf 0.15, 0.26, 0.29, 0.33, 0.38, 0.42, 0.45, 0.53, and Rf 0.58., while peroxidase gave 10 isozymic bands, i.e. Rf 0.06, 0.14, 0.18, 0.22, 0.26, 0.29, 0.32, 0.37, 0.41, and Rf 0.46. Relationship dendrogram indicated that the number of data will affect the grouping based on species similarity; more and more data was increasingly apparent groupings; within these groups there were variations among its members.

  8. Brewing with fractionated barley

    NARCIS (Netherlands)

    Donkelaar, van L.H.G.

    2016-01-01

    Brewing with fractionated barley Beer is a globally consumed beverage, which is produced from malted barley, water, hops and yeast. In recent years, the use of unmalted barley and exogenous enzymes have become more popular because they enable simpler processing and reduced environmental impact. Raw

  9. Brewing with fractionated barley

    NARCIS (Netherlands)

    Donkelaar, van L.H.G.

    2016-01-01

    Brewing with fractionated barley Beer is a globally consumed beverage, which is produced from malted barley, water, hops and yeast. In recent years, the use of unmalted barley and exogenous enzymes have become more popular because they enable simpler processing and reduced environmental impact. Raw

  10. Bioactive phytochemicals in barley

    Directory of Open Access Journals (Sweden)

    Emmanuel Idehen

    2017-01-01

    Full Text Available Epidemiological studies have consistently shown that regular consumption of whole grain barley reduces the risk of developing chronic diseases. The presence of barley fiber, especially β-glucan in whole grain barley, has been largely credited for these health benefits. However, it is now widely believed that the actions of the fiber component alone do not explain the observed health benefits associated with the consumption of whole grain barley. Whole grain barley also contains phytochemicals including phenolic acids, flavonoids, lignans, tocols, phytosterols, and folate. These phytochemicals exhibit strong antioxidant, antiproliferative, and cholesterol lowering abilities, which are potentially useful in lowering the risk of certain diseases. Therefore, the high concentration of phytochemicals in barley may be largely responsible for its health benefits. This paper reviews available information regarding barley phytochemicals and their potential to combat common nutrition-related diseases including cancer, cardiovascular disease, diabetes, and obesity.

  11. Study on The Isozyme of Wild Yak

    Institute of Scientific and Technical Information of China (English)

    许玉德

    2005-01-01

    We have adopted PAGE method to isolate lactic dehydrogenase(LDH)and α-amylase(α-Am)from wild yak's serum and measure the physical-chemical properties of the two components respectively. The results showed that the yak LDH isozyme distributed into five bands. The sequence according to their activity reads as LDH1 > LDH2> LDH3 > LDH4 > LDH5. The subunit B is relatively occupied a dominant position. It got a wide operating pH range, high thermal stability,well denature resistance and a fairly large adaptability threshold on the physical-chemical change of surrounding environment. α-Am isozyme distributed into seven bands and also got a wide operating pH range. α-Am isozyme is high sensitivity to environmental temperature.

  12. Natural polyamines inhibit soybean (Glycine max) lipoxygenase-1, but not the lipoxygenase-2 isozyme.

    Science.gov (United States)

    Maccarrone, M; Baroni, A; Finazzi-Agrò, A

    1998-08-01

    Natural polyamines are shown to inhibit dioxygenase activity of soybean lipoxygenase-1, but they were ineffective toward the lipoxygenase-2 isozyme. The inhibitory power was dependent on the number of basic groups in the molecule, in the order spermine > spermidine > cadaverine >/= putrescine. Both spermidine and spermine acted as uncompetitive inhibitors of lipoxygenase-1 with respect to linoleic acid, the inhibition constants being 2.70 and 0.80 mM, respectively. The inhibitory power apparently correlated with the radical-trapping ability of the polyamines. Spermidine and spermine also inhibited the co-oxidase and peroxidase activities of lipoxygenase-1 and were effective inhibitors of lipoxygenase activity in lentil root protoplasts.

  13. Analysis of Isozyme in Different Leaf Tyoes of Pinellia ternata (Thunb.) Breit%不同类型栽培半夏同工酶分析

    Institute of Scientific and Technical Information of China (English)

    韩凤; 韦波; 封孝兰; 全健; 曹厚强; 梁正杰

    2011-01-01

    采用聚丙烯酰胺凝胶电泳技术,对田间农艺性状表现不同的4种类型半夏叶片中过氧化物酶(POD)和酯酶(EST)的同工酶进行比较,并根据同工酶分析结果探讨半夏种内的亲缘关系.研究表明,过氧化物和酯酶同工酶谱在不同类型的半夏之间有着很高的多态性,除少数共有特征谱带外,各类型之间存在明显的谱带差异.%The isozymes of peroxidase isozyme (POD) and esterase isozyme (EST) in four different types of Pinellia ternate (Thunb.) Breit leaves were compared by using vertical polyacrylamide gel eleetrophoresis, and the relationships among different species of Pinellia ternate (Thunb.) Breit were discussed according to the results of isozyme analysis. The results indicated that the isozyme zymogram of POD and EST in different types of Pinellia ternate (Thunb.) Breit leaves had abundant polyrnorphism, and the differences of POD and EST isozyme zymogram among different types of Pinellia ternate (Thunb.) Breit leaves were obvious except for a few common characteristic zymogram belts.

  14. Characterization of lignin and Mn peroxidases from Phanerochaete chrysosporium. Progress report

    Energy Technology Data Exchange (ETDEWEB)

    1991-12-31

    Long-term objectives are to elucidate the role and mechanism of the various isozymes in lignin biodegradation. Work is described on electrochemical studies on lignin and Mn peroxidases. This study was performed to investigate the structural aspects which confer the lignin and Mn peroxidases with their high reactivity. The experimentally determined redox potential of the Fe{sup 3+}/Fe{sup 2+} couple for the lignin peroxidase isozymes H1, H2, H8 and H10 are very similar, near-130 mV. The redox potential for the Mn peroxidase isozymes H3 and H4 are similar to each other ({minus}88 mV and {minus}95 mV, respectively) and are more positive than the lignin peroxidases. The higher redox potential for the Fe{sup 3+}/Fe{sup 2+} couple is consistent with the heme active site of these fungal peroxidases being more electron deficient. To investigate the accessibility of the heme active site to the substrate which is oxidized [veratryl alcohol and Mn (II)], we investigated whether these substrates had any affect on the redox potential of the heme. The E{sub m7} value for lignin and Mn peroxidases are not affected by their respective substrates, veratryl alcohol and Mn (II). These results suggest that substrates do not directly interact with the ferric heme-iron as axial ligands. This is consistent with the present model for peroxidase catalysis. Suicide inhibitor (1) and nmr studies (2) indicate that the heme-iron of horseradish peroxidase (HRP) is not fully accessible to bulky substrates occur at the periphery of the heme.

  15. Kynurenine Aminotransferase Isozyme Inhibitors: A Review

    Directory of Open Access Journals (Sweden)

    Alireza Nematollahi

    2016-06-01

    Full Text Available Kynurenine aminotransferase isozymes (KATs 1–4 are members of the pyridoxal-5’-phosphate (PLP-dependent enzyme family, which catalyse the permanent conversion of l-kynurenine (l-KYN to kynurenic acid (KYNA, a known neuroactive agent. As KATs are found in the mammalian brain and have key roles in the kynurenine pathway, involved in different categories of central nervous system (CNS diseases, the KATs are prominent targets in the quest to treat neurodegenerative and cognitive impairment disorders. Recent studies suggest that inhibiting these enzymes would produce effects beneficial to patients with these conditions, as abnormally high levels of KYNA are observed. KAT-1 and KAT-3 share the highest sequence similarity of the isozymes in this family, and their active site pockets are also similar. Importantly, KAT-2 has the major role of kynurenic acid production (70% in the human brain, and it is considered therefore that suitable inhibition of this isozyme would be most effective in managing major aspects of CNS diseases. Human KAT-2 inhibitors have been developed, but the most potent of them, chosen for further investigations, did not proceed in clinical studies due to the cross toxicity caused by their irreversible interaction with PLP, the required cofactor of the KAT isozymes, and any other PLP-dependent enzymes. As a consequence of the possibility of extensive undesirable adverse effects, it is also important to pursue KAT inhibitors that reversibly inhibit KATs and to include a strategy that seeks compounds likely to achieve substantial interaction with regions of the active site other than the PLP. The main purpose of this treatise is to review the recent developments with the inhibitors of KAT isozymes. This treatise also includes analyses of their crystallographic structures in complex with this enzyme family, which provides further insight for researchers in this and related studies.

  16. Morphological and isozymic banding pattern study of white grubs (Coleoptera: Melolonthidae as pest of bark crop in mounth Merapi’s slope.

    Directory of Open Access Journals (Sweden)

    SUGIYARTO

    2008-07-01

    Full Text Available White grub (Coleoptera: Melolonthidae is a group of soil pest at any agrosystem., especially at Salak pondoh (Salacca zalacca (Gaert. Voss. crop. The characteristics of this specimen were very crucial to be studied in order to find the exact biocontrol. The aim of this research was to know the characteristics of white grubs (Melolonthidae: Coleoptera based on morphological and isozyme banding patterns. This research was conducted on August - November 2007 at Sleman and Magelang districts for the morphological purposes, while for the isozyme data were conducted at Sub Laboratory Biology, Central Laboratory of Sebelas Maret University Surakarta. Sample was taken by using stratified random sampling method, on five stations. Polyacrylamide gel electrophoresis (PAGE using the vertical type was taken to isozyme analysis. The enzyme used in this research were peroxidase and esterase to detect the isozyme banding patterns. The results showed that there was no morphological variation of white grubs (Melolonthidae: Coleoptera at salak pondoh agroecosystem in Mounth Merapi’s slope. Based on this character, there was one species of white grub found, i.e. Holotrichia javana. There was a genetic variation based on the variation of isozyme banding patterns.

  17. Identification and properties of insect resistance-associated maize anionic peroxidases.

    Science.gov (United States)

    Dowd, Patrick F; Johnson, Eric T; Pinkerton, T Scott

    2010-08-01

    Previous studies with transgenic plants have indicated a tobacco anionic peroxidase can confer enhanced resistance to a variety of insects when expressed in different plant species. Tissue that expresses high levels of this enzyme often browns rapidly when damaged. Maize roots damaged under sterile conditions browned and had an anionic peroxidase induced. When introduced biolistically, maize callus transformants expressing a maize peroxidase gene with a predicted isoelectric point of ca. 5.1 produced browner callus compared to a corresponding beta-glucuronidase (GUS) transformant as callus aged. Higher production of only one isozyme of ca. pI 4.5 was noted. When the callus was fed to two maize pest caterpillar species, growth rates were slower (as reflected by weights) relative to the GUS callus. Based on examination of published information and electrophoretic properties, this gene appears to code for Px11, a peroxidase isozyme that is primarily produced in root tissue and callus. When sequence of the gene in several inbreds was examined, coding variations were noted, and abilities to utilize ferulic and p-coumaric acids differed. These coding differences may influence the ability of corresponding forms of the peroxidase to promote resistance. In addition to potential use in marker assisted breeding, enhanced expression of this anionic peroxidase through breeding or genetic engineering may lead to enhanced insect or disease resistance. Published by Elsevier Ltd.

  18. Root growth inhibition by aluminum is probably caused by cell death due to peroxidase-mediated hydrogen peroxide production.

    Science.gov (United States)

    Simonovicová, M; Huttová, J; Mistrík, I; Siroká, B; Tamás, L

    2004-10-01

    The effect of aluminum on hydrogen peroxide production and peroxidase-catalyzed NADH oxidation was studied in barley roots germinated and grown between two layers of moistened filter paper. Guaiacol peroxidase activity significantly increased after 48 h and was approximately two times higher after 72 h in Al-treated roots. The oxidation of NADH was also significantly increased and, like guaiacol peroxidase activity, it was two times higher in A1-treated roots than in controls. Elevated H2O2 production was observed both 48 and 72 h after the onset of imbibition in the presence of A1. Separation on a cation exchange column allowed the detection of two peaks with NADH peroxidase and H2O2 production activity. However, a difference between control and Al-treated plants was found only in one fraction, in which four times higher guaiacol peroxidase activity and five times higher NADH peroxidase activity were expressed and about three times more H2O2 was produced. One anionic peroxidase and three cationic peroxidases were detected in this fraction by native polyacrylamide gel electrophoresis. The anionic peroxidase was activated in the Al-treated root tips and also oxidized NADH but was detectable only after a long incubation time. Two of the cationic peroxidases were capable of oxidizing NADH and producing a significant amount of H2O2, but only one of these was activated by A1 stress. The role of these peroxidases during A1 stress in barley root tips is discussed.

  19. Response of barley seedlings to oxidative stress generated by treatments with growth hormones

    OpenAIRE

    Zenovia Olteanu; Elena Truta; Lacramiora Oprica; Maria Magdalena Zamfirache

    2009-01-01

    The effects induced by growth hormone regulators on soluble protein level and some oxidoreductases in Hordeum vulgare cv. Madalin seedlings were investigated. The study of superoxide dismutase, catalyse and peroxidase behaviour and of protein synthesis was realized in dynamics to evaluate the response of barley seedlings to oxidative stress generated by exposure to hormone factors. During experiments, peroxidase registered smaller limits of variability than superoxide dismutase an...

  20. Peroxidases in nanostructures

    Directory of Open Access Journals (Sweden)

    Ana Maria eCarmona-Ribeiro

    2015-09-01

    Full Text Available Peroxidases are enzymes catalyzing redox reactions that cleave peroxides. Their active redox centers have heme, cysteine thiols, selenium, manganese and other chemical moieties. Peroxidases and their mimetic systems have several technological and biomedical applications such as environment protection, energy production, bioremediation, sensors and immunoassays design and drug delivery devices. The combination of peroxidases or systems with peroxidase-like activity with nanostructures such as nanoparticles, nanotubes, thin films, liposomes, micelles, nanoflowers, nanorods and others is often an efficient strategy to improve catalytic activity, targeting and reusability.

  1. Peroxidase(s) in Environment Protection

    Science.gov (United States)

    Bansal, Neelam; Kanwar, Shamsher S.

    2013-01-01

    Industrial discharges of untreated effluents into water bodies and emissions into air have deteriorated the quality of water and air, respectively. The huge amount of pollutants derived from industrial activities represents a threat for the environment and ecologic equilibrium. Phenols and halogenated phenols, polycyclic aromatic hydrocarbons (PAH), endocrine disruptive chemicals (EDC), pesticides, dioxins, polychlorinated biphenyls (PCB), industrial dyes, and other xenobiotics are among the most important pollutants. Peroxidases are enzymes that are able to transform a variety of compounds following a free radical mechanism, thereby yielding oxidized or polymerized products. The peroxidase transformation of these pollutants is accompanied by a reduction in their toxicity, due to loss of biological activity, reduction in the bioavailability, or the removal from aqueous phase, especially when the pollutant is found in water. The review describes the sources of peroxidases, the reactions catalyzed by them, and their applications in the management of pollutants in the environment. PMID:24453894

  2. Resolution of brewers' yeast pyruvate decarboxylase into two isozymes.

    Science.gov (United States)

    Kuo, D J; Dikdan, G; Jordan, F

    1986-03-01

    A novel purification method was developed for brewers' yeast pyruvate decarboxylase (EC 4.1.1.1) that for the first time resolved the enzyme into two isozymes on DEAE-Sephadex chromatography. The isozymes were found to be distinct according to sodium dodecyl sulfate polyacrylamide gel electrophoresis: the first one to be eluted gave rise to one band, the second to two bands. The isozymes were virtually the same so far as specific activity, KM, inhibition kinetics and irreversible binding properties by the mechanism-based inhibitor (E)-4-(4-chlorophenyl)-2-oxo-3-butenoic acid are concerned. This finding resolves a longstanding controversy concerning the quaternary structure of this enzyme.

  3. General and Versatile Autoinhibition of PLC Isozymes

    Energy Technology Data Exchange (ETDEWEB)

    Hicks, Stephanie N.; Jezyk, Mark R.; Gershburg, Svetlana; Seifert, Jason P.; Harden, T. Kendall; Sondek, John (UNC)

    2008-10-31

    Phospholipase C (PLC) isozymes are directly activated by heterotrimeric G proteins and Ras-like GTPases to hydrolyze phosphatidylinositol 4,5-bisphosphate into the second messengers diacylglycerol and inositol 1,4,5-trisphosphate. Although PLCs play central roles in myriad signaling cascades, the molecular details of their activation remain poorly understood. As described here, the crystal structure of PLC-{beta}2 illustrates occlusion of the active site by a loop separating the two halves of the catalytic TIM barrel. Removal of this insertion constitutively activates PLC-{beta}2 without ablating its capacity to be further stimulated by classical G protein modulators. Similar regulation occurs in other PLC members, and a general mechanism of interfacial activation at membranes is presented that provides a unifying framework for PLC activation by diverse stimuli.

  4. Isozyme-based genetic fingerprinting of Manihot sp

    African Journals Online (AJOL)

    Prof. Ogunji

    1973-06-22

    Jun 22, 1973 ... Development of Crop & Soil Science, University of Port Harcourt P.M.B. 5323 ... In order to use isozyme techniques successfully for cultivar identification, a particular ..... The Principles and Practice of numerical classification.

  5. Malting barley BRS Borema

    Directory of Open Access Journals (Sweden)

    Euclydes Minella

    2006-01-01

    Full Text Available BRS Borema is an early maturing, two-rowed spring barley registered in 2003 for commercial production inSouthern Brazil, bred by Embrapa Trigo. It combines good yield potential with superior malting quality and a reasonable levelof disease (net blotch, powdery mildew, leaf rust resistance. It is well-adapted to all major production regions of maltingbarley in Brazil.

  6. Effects of calcium ion concentration on starch hydrolysis of barley α-amylase isozymes

    DEFF Research Database (Denmark)

    Yuk, Jeong-Bin; Choi, Seung-Ho; Lee, Tae-Hee

    2008-01-01

    in glucose, maltose, or maltotriose. Against insoluble blue starch, AMY1 showed the highest activity at 0.1-5 mM calcium concentration, whereas 15-20 mM was optimal for AMY2. On the hydrolysis of soluble starch, unexpectedly, there was no significant difference between AMYs with increase of calcium. However...

  7. Arabidopsis thaliana peroxidase N

    DEFF Research Database (Denmark)

    Mirza, Osman Asghar; Henriksen, A; Ostergaard, L

    2000-01-01

    The structure of the neutral peroxidase from Arabidopsis thaliana (ATP N) has been determined to a resolution of 1.9 A and a free R value of 20.5%. ATP N has the expected characteristic fold of the class III peroxidases, with a C(alpha) r.m.s.d. of 0.82 A when compared with horseradish peroxidase C...... (HRP C). HRP C is 54% identical to ATP N in sequence. When the structures of four class III plant peroxidases are superimposed, the regions with structural differences are non-randomly distributed; all are located in one half of the molecule. The architecture of the haem pocket of ATP N is very similar...... to that of HRP C, in agreement with the low small-molecule substrate specificity of all class III peroxidases. The structure of ATP N suggests that the pH dependence of the substrate turnover will differ from that of HRP C owing to differences in polarity of the residues in the substrate-access channel. Since...

  8. Changes of Soluble Protein, Peroxidase Activity and Distribution During Regeneration After Girdling in Eucommia ulmoides

    Institute of Scientific and Technical Information of China (English)

    HOUHong-Wei; Kalima-N'KomaMWANGE; WANGYa-Qing; CUIKe-Ming

    2004-01-01

    Peroxidases are known to play important roles in plant wound healing. Biochemical analysisand histochemical localization techniques were used to assess changes and distribution of peroxidases inthe recovering bark after girdling in Eucommia ulmoides Oliv. Between 4 and 21 days after girdling (DAG),peroxidases activity in the girdled trees significantly increased by 30-40 times over that in ungirdled trees.During the whole bark recovery process (from 0 to 63 DAG), the peroxidase signal was not found in thetissue regions subjected to intense cell division activity (regenerating cambial zone and phellogen). However,high peroxidase activity was detected in the callus, cortex-like, mature phloem and xylem. Interestingly, itwas shown that, in maturing xylem and phloem cells, there was respectively an inward and outwardperoxidase activity gradient on both sides of the cambium zone. An isoelectric-focusing electrophoresis ofthe extracted protein displayed two isozyme bands of peroxidase: POD Ⅰ and POD Ⅱ. POD Ⅰ was onlydetected in the xylem fraction and could play a role in xylem differentiation. POD Ⅱ was only identified inthe recovering bark portion and could be more engaged in bark regeneration process. A relationshipbetween IAA and peroxidase is also discussed.

  9. HEALTH BENEFITS OF BARLEY

    Directory of Open Access Journals (Sweden)

    Akula Annapurna

    2013-09-01

    Full Text Available Prevalence of lifestyle diseases is increasing day by day. Mostly the younger generation do not have much awareness about healthy nutritional supplements. One such important cereal grain not used mostly by youngsters is barley It is a good old grain with so many health benefits like weight reduction, decreasing blood pressure, blood cholesterol, blood glucose in Type 2 diabetes and preventing colon cancer. It is easily available and cheap grain. It contains both soluble and insoluble fiber, protein, vitamins B and E, minerals selenium, magnesium and iron, copper, flavonoids and anthocynins. Barley contains soluble fiber, beta glucan binds to bile acids in the intestines and thereby decreasing plasma cholesterol levels. Absorbed soluble fiber decreases cholesterol synthesis by liver and cleansing blood vessels. Insoluble fiber provides bulkiness in the intestines, thereby satiety. decreased appetite. It promotes intestinal movements relieving constipation, cleansing colonic harmful bacteria and reduced incidence of colonic cancer. It is a good source of niacin ,reducing LDL levels and increasing HDL levels. Selenium and vitamin E providing beneficial antioxidant effects. Magnesium, a cofactor for many carbohydrate metabolism enzymes and high fiber content contributes for its blood glucose reducing effect in Type 2 diabetes. It is having good diuretic activity and is useful in urinary tract infections. Barley contains gluten, contraindicated in celiac disease.

  10. Glutamic oxaloacetic transaminase isozymes from rat liver. Purification and physicochemical characterization.

    Science.gov (United States)

    Huynh, Q K; Sakakibara, R; Watanabe, T; Wada, H

    1980-07-01

    Glutamic oxaloacetic transaminase isozymes were purified simultaneously to homogeneity from rat liver with high yields. Three subforms of mitochondrial isozyme and three subforms of cytosolic isozyme were separated by chromatography on CM-Sephadex and electrophoresis on polyacrylamide gel. The general enzymatic properties of the purified isozymes such as their kinetic parameters, isoelectric points, molecular weights, amino acid compositions, NH2-terminal amino acid sequences and COOH-terminal amino acids were determined. Most of these properties of the isozymes are similar to those of the corresponding isozymes from other sources, such as rat brain and pig and human heart. In amino acid compositions, cytosolic isozyme from rat liver has more proline and glycine and less arginine, threonine and leucine than pig heart cytosolic isozyme; the mitochondrial isozyme has more glutamic acid and glycine and less serine than the corresponding pig heart isozyme. The NH2-terminal amino acid sequences of GOT isozymes from rat liver were identical with those of the GOT isozymes from pig heart up to the 10th residues except for the 5th residues. The subforms of mitochondrial isozyme from rat liver were generated on storage at 4 degrees C for 4-8 weeks.

  11. Novel Applications of Peroxidase

    Science.gov (United States)

    Rob, Abdul; Ball, Andrew S.; Tuncer, Munir; Wilson, Michael T.

    1997-02-01

    The article entitled "Novel Biocatalysts Will Work Even Better for Industry" published recently in this Journal (1) was informative and interesting. However it touched only briefly on the application of peroxidase as catalyst. Here, we would like to mention in more detail the novel applications of peroxidase in agricultural, paper pulp, water treatment, pharmaceutical, and medical situations. Firstly, the peroxidase isolated from Phanerochaete chyrosporium has been shown to detoxify herbicides such as atrazine to less toxic compounds and would certainly find potential application in agriculture (2). Secondly, the peroxidase produced by Streptomyces thermoviolaceus may find application in the paper pulp industry as a delignifying agent (3). Thirdly, it has been shown that extracellular peroxidase produced by Streptomyces avermitilis can remove the intense color from paper-mill effluent obtained after semichemical alkaline pulping of wheat straw (4), and thus this enzyme might find application as a catalyst in water treatment plants. Fourthly, the heme-containing horseradish peroxidase enzyme has been exploited in several diagnostic applications in pharmaceutics and medicine, such as the detection of human immunodeficiency virus and cystic fibrosis (5-10). Finally, recent work from our laboratory has suggested that thermophilic nonheme peroxidase produced by Thermomonospora fusca BD25 may find medical use in the diagnosis of myocardial infarction (11, 12). Literature Cited 1. Wiseman, A. J. Chem. Educ. 1996, 73, 55-58. 2. Mougin, C. Appl. Environ. Microbiol. 1994, 60, 705-708. 3. McCarthy A. J.; Peace, W.; Broda, P. Appl. Microbiol. Technol. 1985, 23, 238-244. 4. Hernandez, M; Rodriguez J; Soliveri, J; Copa, J. L; Perez, M. I; Arias, M. E. Appl. Environ. Microbiol. 1994, 60, 3909-3913. 5. Hopfer, S. M.; Aslanzadeh, J. Ann. Clin. Lab. Sci. 1995, 25, 475-480. 6. Suzuki, K; Iman, M. J. Virol. Methods 1995, 55, 347-356. 7. Nielsen, K. J. Immunoassay 1995, 16, 183-197. 8

  12. Curcumin Prevents Aflatoxin B1 Hepatoxicity by Inhibition of Cytochrome P450 Isozymes in Chick Liver

    Directory of Open Access Journals (Sweden)

    Ni-Ya Zhang

    2016-11-01

    Full Text Available This study was designed to establish if Curcumin (CM alleviates Aflatoxin B1 (AFB1-induced hepatotoxic effects and to determine whether alteration of the expression of cytochrome P450 (CYP450 isozymes is involved in the regulation of these effects in chick liver. One-day-old male broilers (n = 120 were divided into four groups and used in a two by two factorial trial in which the main factors included supplementing AFB1 (< 5 vs. 100 μg/kg and CM (0 vs. 150 mg/kg in a corn/soybean-based diet. Administration of AFB1 induced liver injury, significantly decreasing albumin and total protein concentrations and increasing alanine aminotransferase and aspartate aminotransferase activities in serum, and induced hepatic histological lesions at week 2. AFB1 also significantly decreased hepatic glutathione peroxidase, catalase, and glutathione levels, while increasing malondialdehyde, 8-hydroxydeoxyguanosine, and exo-AFB1-8,9-epoxide (AFBO-DNA concentrations. In addition, the mRNA and/or activity of enzymes responsible for the bioactivation of AFB1 into AFBO—including CYP1A1, CYP1A2, CYP2A6, and CYP3A4—were significantly induced in liver microsomes after 2-week exposure to AFB1. These alterations induced by AFB1 were prevented by CM supplementation. Conclusively, dietary CM protected chicks from AFB1-induced liver injury, potentially through the synergistic actions of increased antioxidant capacities and inhibition of the pivotal CYP450 isozyme-mediated activation of AFB1 to toxic AFBO.

  13. Isozyme patterns and protein profiles in neuromuscular disorders.

    Science.gov (United States)

    Edwards, Y H; Tipler, T D; Morgan-Hughes, J A; Neerunjun, J S; Hopkinson, D A

    1982-06-01

    The isozyme patterns of six different enzymes and the polypeptide profiles of soluble proteins have been examined in muscle biopsy specimens from 74 patients with a wide variety of neuromuscular disorders. About half of the samples showed unusual features in at least one, and often several, of the enzymes and proteins tested. The extent of the biochemical abnormalities was roughly proportional to the severity of the disorders. In all cases the unusual isozymes and polypeptide profiles seemed to reflect a reversion to the fetal pattern of gene expression. However, this change appeared to occur in extant muscle and was not dependent on the appearance of new muscle fibres. Among the enzymes, phosphoglycerate mutase followed by creatine kinase appeared to be the most sensitive index of muscle disorder. The extent of the change in the muscle creatine kinase isozyme pattern was not correlated with the levels of serum creatine kinase activity.

  14. Genomic Prediction in Barley

    DEFF Research Database (Denmark)

    Edriss, Vahid; Cericola, Fabio; Jensen, Jens D

    2015-01-01

    Genomic prediction uses markers (SNPs) across the whole genome to predict individual breeding values at an early growth stage potentially before large scale phenotyping. One of the applications of genomic prediction in plant breeding is to identify the best individual candidate lines to contribute...... to next generation. The main goal of this study was to see the potential of using genomic prediction in a commercial Barley breeding program. The data used in this study was from Nordic Seed company which is located in Denmark. Around 350 advanced lines were genotyped with 9K Barely chip from Illumina...

  15. Differential Antioxidative Responses to Water Deficit Among four Barley (Hordeum vulgare L. Genotypes

    Directory of Open Access Journals (Sweden)

    Z Amini

    2013-08-01

    Full Text Available Future climate changes are expected to increase risks of drought, which already represent the most common stress factor for stable barley (Hordeum vulgare L. production in Iran. Up to now, extensive research projects have been done to study effects of drought stress on the antioxidant enzyme activity. While there is a few works of such studies on the field condition. In order to study of water deficit effects on the antioxidant enzymes activities as a secondary stress, we evaluate the effects of mild and severe drought stress on activities of antioxidative enzymes including superoxide dismutases, ascorbate peroxidase, catalase and peroxidase, among four barley genotypes, differing in the capacity to maintain the grain yield under drought condition during beginning on anthesis, kernel watery ripe and late milk stages under field condition. Results showed that drought increased the activity of antioxidant enzymes in all genotypes. At beginning of anthesis, POX activity of Q22 was higher than it in other genotypes ( P

  16. Isozyme Analysis of Spotted Halibut Verasper variegatus Temminck et Schlegel

    Institute of Scientific and Technical Information of China (English)

    LIU Manhong; GAO Tianxiang; ZHANG Xiumei; CHEN Siqing

    2005-01-01

    To investigate the tissue-specificities of isozymes and the genetic structure of wild spotted halibut ( Verasper vari gatus) population, horizontal starch gel electrophoresis was performed on 45 individuals collected in part of the Yellow Sea.The performances of 17 isozymes in 8 kinds of tissues or organs were screened preliminarily in a TC-7.0 buffer system. The results showed that the screened isozymes displayed remarkable tissue-specificities. Finally, 14 enzymes (AAT, ADH, EST,GPI, G3PDH, IDHP, LAP, LDH, MDH, MPI, PGDH, PGM, SDH and SOD) and 4 kinds of tissues (eye, skeleton mus cle, liver and heart) were selected for genetic analysis. Fourteen isozymes are encoded by 20 loci, and 9 of them are poly morphic. The polymorph ic loci are AAT-1*, GPI-2*, G3PDH*, IDHP-1*, LDH*, MPI*, PGM-1*, PGM-2* and SDH* , and the proportion of polymorphic loci is 0.4500 (P0.99). The mean values of observed and expected heterozygosities are 0.027 8 and 0.026 5, respectively and the average effective number of alleles is 1.067 5.

  17. Acrolein-detoxifying isozymes of glutathione transferase in plants.

    Science.gov (United States)

    Mano, Jun'ichi; Ishibashi, Asami; Muneuchi, Hitoshi; Morita, Chihiro; Sakai, Hiroki; Biswas, Md Sanaullah; Koeduka, Takao; Kitajima, Sakihito

    2017-02-01

    Acrolein is a lipid-derived highly reactive aldehyde, mediating oxidative signal and damage in plants. We found acrolein-scavenging glutathione transferase activity in plants and purified a low K M isozyme from spinach. Various environmental stressors on plants cause the generation of acrolein, a highly toxic aldehyde produced from lipid peroxides, via the promotion of the formation of reactive oxygen species, which oxidize membrane lipids. In mammals, acrolein is scavenged by glutathione transferase (GST; EC 2.5.1.18) isozymes of Alpha, Pi, and Mu classes, but plants lack these GST classes. We detected the acrolein-scavenging GST activity in four species of plants, and purified an isozyme showing this activity from spinach (Spinacia oleracea L.) leaves. The isozyme (GST-Acr), obtained after an affinity chromatography and two ion exchange chromatography steps, showed the K M value for acrolein 93 μM, the smallest value known for acrolein-detoxifying enzymes in plants. Peptide sequence homology search revealed that GST-Acr belongs to the GST Tau, a plant-specific class. The Arabidopsis thaliana GST Tau19, which has the closest sequence similar to spinach GST-Acr, also showed a high catalytic efficiency for acrolein. These results suggest that GST plays as a scavenger for acrolein in plants.

  18. Alleviation of Al Toxicity in Barley by Addition of Calcium

    Institute of Scientific and Technical Information of China (English)

    GUO Tian-rong; CHEN Ying; ZHANG Yan-hua; JIN Ye-fei

    2006-01-01

    The potential mechanism by which Ca alleviates Al toxicity was investigated in barley seedlings. It was found that 100 μM Al-alone treatment inhibited barley plant growth and thereby reduced shoot height and root length, and dry weights of root, shoot and leaf; promoted Al accumulation but inhibited Ca absorption in plant tissues; and induced an increase in the activities of superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) and in the level of lipid peroxidation (MDA content) in leaves. Except for the increase in Ca concentration in plant tissues, treatment with 0.5 mM Ca in the absence of Al had less effect on the above-mentioned parameters, compared with the control. Addition of Ca efficiently reduced Al toxicity, which is reflected by the promotion of plant growth, reduction in Al concentration and MDA content,increase in Ca concentration and in SOD, POD, and CAT activities compared with the Al-alone-treatment; with increase in Ca level (3.0 mM), the ameliorative effect became more dominant. This indicated that the alleviation of aluminum toxicity in barley seedlings with Ca supplementation could be associated with less absorption of Al and the enhancement of the protective ability of the cell because of increased activity of the antioxidative enzyme.

  19. Interisland evolution of Trimeresurus flavoviridis venom phospholipase A(2) isozymes.

    Science.gov (United States)

    Chijiwa, Takahito; Yamaguchi, Yoko; Ogawa, Tomohisa; Deshimaru, Masanobu; Nobuhisa, Ikuo; Nakashima, Kinichi; Oda-Ueda, Naoko; Fukumaki, Yasuyuki; Hattori, Shosaku; Ohno, Motonori

    2003-03-01

    Trimeresurus flavoviridis snakes inhabit the southwestern islands of Japan. A phospholipase A(2) (PLA(2)), named PL-Y, was isolated from Okinawa T. flavoviridis venom and its amino acid sequence was determined from both protein and cDNA. PL-Y was unable to induce edema. In contrast, PLA-B, a PLA(2) from Tokunoshima T. flavoviridis venom, which is different at only three positions from PL-Y, is known to induce edema. A new PLA(2), named PLA-B', which is similar to PLA-B, was cloned from Amami-Oshima T. flavoviridis venom gland. Three T. flavoviridis venom basic [Asp(49)]PLA(2) isozymes, PL-Y (Okinawa), PLA-B (Tokunoshima), and PLA-B' (Amami-Oshima), are identical in the N-terminal half but have one to four amino acid substitutions in the beta1-sheet and its vicinity. Such interisland sequence diversities among them are due to isolation in the different environments over 1 to 2 million years and appear to have been brought about by natural selection for point mutation in their genes. Otherwise, a major PLA(2), named PLA2, ubiquitously exists in the venoms of T. flavoviridis snakes from the three islands with one to three synonymous substitutions in their cDNAs. It is assumed that the PLA2 gene is a prototype among T. flavoviridis venom PLA(2) isozyme genes and has hardly undergone nonsynonymous mutation as a principal toxic component. Phylogenetic analysis based on the amino acid sequences revealed that T. flavoviridis PLA(2) isozymes are clearly separated into three groups, PLA2 type, basic [Asp(49)]PLA(2) type, and [Lys(49)]PLA(2) type. Basic [Asp(49)]PLA(2)-type isozymes may manifest their own particular toxic functions different from those of the isozymes of the PLA2 type and [Lys(49)]PLA(2) type.

  20. Barley Transformation Using Biolistic Techniques

    Science.gov (United States)

    Harwood, Wendy A.; Smedley, Mark A.

    Microprojectile bombardment or biolistic techniques have been widely used for cereal transformation. These methods rely on the acceleration of gold particles, coated with plasmid DNA, into plant cells as a method of directly introducing the DNA. The first report of the generation of fertile, transgenic barley plants used biolistic techniques. However, more recently Agrobacterium-mediated transformation has been adopted as the method of choice for most cereals including barley. Biolistic procedures are still important for some barley transformation applications and also provide transient test systems for the rapid checking of constructs. This chapter describes methods for the transformation of barley using biolistic procedures and also highlights the use of the technology in transient assays.

  1. Caracterização isozimática e atividade de peroxidase em folhas de plantas hiperídrica, intermediária e normal de Bidens pilosa L. mantidas in vitro Isoezymatic characterization and peroxidase activity in leaves of hyperhydric, intermediary and normal plants of Bidens pilosa L. grown in vitro

    Directory of Open Access Journals (Sweden)

    José Emílio Zanzirolani de Oliveira

    2008-02-01

    Full Text Available Foram caracterizadas as plantas: hiperídrica, intermediária e normal de um clone de Bidens pilosa mantido em cultivo in vitro por meio de isozimas e da atividade de peroxidase. Empregando-se a eletroforese em géis de amido a 12%, testou-se seis isozimas, sendo detectado polimorfismo em peroxidase e fosfatase ácida, permitindo caracterizar cada tipo de planta. Não houve polimorfismo em fosfogluco isomerase, fosfoglucomutase, glutamato oxaloacetato transaminase e malato desidrogenase. A atividade da peroxidase foi maior nas plantas hiperídricas e intermediárias. Conclui-se que a variabilidade enzimática tem potencial como marcador de hiperidricidade em plantas mantidas in vitro.Activity of peroxidase (EC 1.11.1.7 and isozymes analysis of a Bidens pilosa clone maintained in vitro culture were characterized in hyperhydric, intermediary and normal plants. Electrophorese in starch gels (12% of six isozymes systems was tested, polymorphisms in peroxidase and acid phosphatase (EC 3.1.3.2 were detected. There was absence of polymorphism in phosphoglucoisomerase (EC 5.3.1.9, phosphoglucomutase (EC 5.4.2.2, glutamate oxaloacetate transaminase (EC 2.6.1.1 and malate dehydrogenase (EC 1.1.1.37. Comparing the activity of peroxidase enzyme, it was higher in hyperhydric and intermediary plants in relation to normal ones. Enzymatic variability is a potential tool as hyperhydricity marker in plants grown in vitro.

  2. Functional proteomics of barley and barley chloroplasts – strategies, methods and perspectives

    DEFF Research Database (Denmark)

    Petersen, Jørgen; Rogowska-Wrzesinska, Adelina; Jensen, Ole Nørregaard

    2013-01-01

    tolerance, micronutrient utilization, and photosynthesis in barley. In the present review we present the current state of proteomics research for investigations of barley chloroplasts, i.e., the organelle that contain the photosynthetic apparatus in the plant. We describe several different proteomics...... strategies and discuss their applications in characterization of the barley chloroplast as well as future perspectives for functional proteomics in barley research....

  3. Isozyme variation within and among populations of Microsporum species.

    Science.gov (United States)

    Simpanya, M F; Jarvis, B D; Baxter, M

    1998-12-01

    Isozyme variation among 54 isolates of Microsporum canis, 18 Microsporum cookei isolates and two Diheterospora isolates were studied using starch gel electrophoresis. Of eight enzymes examined, four were polymorphic (EST, G6P, MDH and PEP), having from two to four electrophoretic forms. Within each species, consistent and reproducible isozyme patterns of the eight enzyme systems were obtained. Phenotypic diversity (H) in M. canis was higher than in M. cookei (H = 0.459 and H = 0.408 respectively), but phenotypic differentiation of M. canis isolates from different geographical regions (Auckland, Wellington and Palmerston North, New Zealand) was low, with a proportion of total diversity (Gst) of 0.151 found among the localities. The results suggest that the isolates of M. canis from different geographical regions are closely related, supporting the theory of a common lineage.

  4. Variability among inbred lines and RFLP mapping of sunflower isozymes

    Directory of Open Access Journals (Sweden)

    Carrera Alicia D.

    2002-01-01

    Full Text Available Eight isozyme systems were used in this study: acid phosphatase (ACP, alcohol dehydrogenase (ADH, esterase (EST, glutamate dehydrogenase (GDH, malate dehydrogenase (MDH, phosphoglucoisomerase (PGI, 6-phosphogluconate dehydrogenase (PGD, and phosphoglucomutase (PGM. The polymorphism of these enzyme systems was studied in 25 elite inbred lines. A total of 19 loci were identified, but only eight of them were polymorphic in the germplasm tested. The polymorphic index for the eight informative markers ranged from 0.08 to 0.57, with a mean value of 0.36. Five isozyme loci were mapped in F2:3 populations with existing RFLP data. Est-1, Gdh-2 and Pgi-2 were mapped to linkage groups 3, 14 and 9, respectively. As in previous reports, an ACP locus and a PGD locus were found to be linked, both located in linkage group 2 of the public sunflower map.

  5. Isozymes analysis of the golden cuttlefish Sepia esculenta (Cephalopoda: Sepiidae)

    Science.gov (United States)

    Zheng, Xiaodong; Zhao, Jianmin; Xiao, Shu; Wang, Rucai; Wang, Shidang; Zhou, Weiwu

    2004-04-01

    Thirty-nine isozymes in four tissues (mantle muscle, buccal bulb muscle, eye and liver) of Sepia esculenta were screened for enzymatic analysis using starch gel electrophoretic technique. Eighteen enzymes (G3PDH, LDH, MDH, MEP, IDHP, PGDH, GRS, NP, AAT, CK, AK, EST, ALP, ACP, FBP, MPI, GPI and PGM) show strong activities and good convergence in zymogram. They are proved to be suitable genetic markers in Sepia esculenta. Among the tissues used, mantle muscle is the best for electrophoretic analysis of isozymes. Eye and liver are fairly good for some special enzymes, such as LDH, EST, MPI, etc. Twenty-six loci are detected. The proportion of polymorphic loci is 0.115 in the Qingdao sample and 0.153 in the Rizhao sample (Pvalues of the observed and expected heterozygosity per locus of Qingdao sample are 0.016 and 0.017, while those of the Rizhao sample are 0.023 and 0.025 respectively.

  6. Secretion, purification, and characterisation of barley alpha-amylase produced by heterologous gene expression in Aspergillus niger.

    Science.gov (United States)

    Juge, N; Svensson, B; Williamson, G

    1998-04-01

    Efficient production of recombinant barley alpha-amylase has been achieved in Aspergillus niger. The cDNA encoding alpha-amylase isozyme 1 (AMY1) and its signal peptide was placed under the control of the Aspergillus nidulans glyceraldehyde-3-phosphate dehydrogenase (gpd) promoter and the A. nidulans trpC gene terminator. Secretion yields up to 60 mg/l were obtained in media optimised for alpha-amylase activity and low protease activity. The recombinant AMY1 (reAMY1) was purified to homogeneity and found to be identical to native barley AMY1 with respect to size, pI, and immunoreactivity. N-terminal sequence analysis of the recombinant protein indicated that the endogenous plant signal peptide is correctly processed in A. niger. Electrospray ionisation/mass spectrometry gave a molecular mass for the dominant form of 44,960 Da, in accordance with the loss of the LQRS C-terminal residues; glycosylation apparently did not occur. The activities of recombinant and native barley alpha-amylases are very similar towards insoluble and soluble starch as well as 2-chloro-4-nitrophenol beta-D-maltoheptaoside and amylose (degree of polymerisation = 17). Barley alpha-amylase is the first plant protein efficiently secreted and correctly processed by A. niger using its own signal sequence.

  7. Characterization of thermostable β-amylase isozymes from Lactobacillus fermentum.

    Science.gov (United States)

    Kocabay, Samet; Çetinkaya, Serap; Akkaya, Birnur; Yenidünya, Ali Fazil

    2016-12-01

    A strain of Lactobacillus fermentum producing two isozymes of a 20kDa β-amylase was isolated from the faecal sample of a newborn. The starin was identified by sequencing its 16S rRNA gene. The two β-amylase isozymes were resolved and visualized by two dimensional protein gel electrophoresis (2-D gel electrophoresis). Some of the physical and biochemical properties of the enzymes were characterized. The β-amylase displayed two optimum pH s, 5.0 and 10.0 and two optimum temperatures, 45°C and 37°C, respectively. The isozymes hydrolyzed different substrates: glycogen at pH 5.0, and corn starch at pH 10.0. The activity did not require Ca(2+), though the activity at pH 10.0 was enhanced in the presence of 5.0mM and 10.0mM CaCl2, 110% and 130%, respectively. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Peroxidase activity in plant leaves exposed to gaseous HCl or ozone

    Energy Technology Data Exchange (ETDEWEB)

    Endress, A.G.; Suarez, S.J.; Taylor, O.C.

    1980-01-01

    Previous studies of plants subjected to air pollutant stress indicated that increased total peroxidase (PRO) activity and/or altered PRO isozyme patterns are frequently induced. Phaseolus vulgaris var. UI 111 and Lycopersicon esculentum var. Heinz 1350 plants exposed to either gaseous hydrogen chloride (HCl) for 20 min or ozone (O/sub 3/) for 1 h were employed to determine if (1) total PRO activity increased following pollutant stress, (2) increased PRO activity was associated with non-necrotic injury, and (3) PRO isozyme patterns were altered after pollutant stress. Non-necrotic foliar macroscopic injury was dependent on pollutant concentration in the bean-O/sub 3/, tomato-O/sub 3/, and bean-HCl combinations, but not for the combination of tomato-HCl where there was little injury. Total PRO activity increased for all combinations. In only one (bean-O/sub 3/), however, was elevated PRO activity statistically related to pollutant concentration. In the others, only the post-exposure time interval was related to PRO activity. No differences of isozyme pattern were discerned between PRO samples obtained from tomato exposed to charcoal-filtered air and either O/sub 3/ or HCl. The PRO isozyme pattern of samples obtained from bean varied with imposed stress and sampling time. The results obtained indicate that PRO levels may be elevated by subjection to air pollutant stress and that PRO activity is sensitive to the internal physiological conditions of the plants. Its extreme sensitivity, however, precludes its use as a reliable indicator of pollutant stress.

  9. CO-FERMENTATION OF KOCHO WITH BARLEY

    African Journals Online (AJOL)

    improved protein content in which kocho and barley flour were fermented for 96 hrs with barley flour. ... involving co-fermentation of kocho and barley flour for the production of nutritionally improved ..... chickpeas. J. Food Sci. 44:234-236.

  10. List of isozyme loci - RGP gmap98 | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available List Contact us RGP gmap98 List of isozyme loci Data detail Data name List of isozyme loci DOI 10.18908/lsdb...he present high-density linkage map, and that were putatively identified as isozyme genes. Data file File name: rgp_gmap98_iso...gmap98/LATEST/rgp_gmap98_isozyme_loci.zip File size: 611 B Simple search URL http://togodb.biosciencedbc.jp/...0001 were considered as functionally identical clones. And we have selected the ones that hit the isozyme ge...his Database Database Description Download License Update History of This Database Site Policy | Contact Us List of isozyme loci - RGP gmap98 | LSDB Archive ...

  11. Isozyme, ISSR and RAPD profiling of genotypes in marvel grass (Dichanthium annulatum).

    Science.gov (United States)

    Saxena, Raghvendra; Chandra, Amaresh

    2010-11-01

    Genetic analysis of 30 accessions of marvel grass (Dichanthium annulatum Forsk.), a tropical range grass collected from grasslands and open fields of drier regions, was carried out with the objectives of identifying unique materials that could be used in developing the core germplasm for such regions as well as to explore gene (s) for drought tolerance. Five inter-simple sequence repeat (ISSR) primers [(CA)4, (AGAC), (GACA) 4; 27 random amplified polymorphic DNA (RAPD) and four enzyme systems were employed in the present study. In total, ISSR yielded 61 (52 polymorphic), RAPD 269 (253 polymorphic) and enzyme 55 isozymes (44 polymorphic) bands. The average polymorphic information content (PIC) and marker index (MI) across all polymorphic bands of 3 markers systems ranged from 0.419 to 0.480 and 4.34 to 5.25 respectively Dendrogram analysis revealed three main clusters with all three markers. Four enzymes namely esterase (EST), polyphenoloxidase (PPO), peroxidase (PRX) and superoxide dismutase (SOD) revealed 55 alleles from a total of 16 enzyme-coding loci. Of these, 14 loci and 44 alleles were polymorphic. The mean number of alleles per locus was 3.43. Mean heterozygosity observed among the polymorphic loci ranged from 0.406 (SOD) to 0.836 (EST) and accession wise from 0.679 (1G3108) to 0.743 (IGKMD-10). Though there was intermixing of few accessions of one agro-climatic region to another largely groupings of accessions were with their regions of collections. Bootstrap analysis at 1000 iterations also showed large numbers of nodes (11 to 17) having strong clustering (> 50 bootstrap values) in all three marker systems. The accessions of the arid and drier regions forming one cluster are assigned as distinct core collection of Dichanthium and can be targeted for isolation of gene (s) for drought tolerance. Variations in isozyme allele numbers and high PIC (0.48) and MI (4.98) as observed with ISSR markers indicated their usefulness for germplasm characterization.

  12. Alanine aminotransferase controls seed dormancy in barley

    Science.gov (United States)

    Sato, Kazuhiro; Yamane, Miki; Yamaji, Nami; Kanamori, Hiroyuki; Tagiri, Akemi; Schwerdt, Julian G.; Fincher, Geoffrey B.; Matsumoto, Takashi; Takeda, Kazuyoshi; Komatsuda, Takao

    2016-01-01

    Dormancy allows wild barley grains to survive dry summers in the Near East. After domestication, barley was selected for shorter dormancy periods. Here we isolate the major seed dormancy gene qsd1 from wild barley, which encodes an alanine aminotransferase (AlaAT). The seed dormancy gene is expressed specifically in the embryo. The AlaAT isoenzymes encoded by the long and short dormancy alleles differ in a single amino acid residue. The reduced dormancy allele Qsd1 evolved from barleys that were first domesticated in the southern Levant and had the long dormancy qsd1 allele that can be traced back to wild barleys. The reduced dormancy mutation likely contributed to the enhanced performance of barley in industrial applications such as beer and whisky production, which involve controlled germination. In contrast, the long dormancy allele might be used to control pre-harvest sprouting in higher rainfall areas to enhance global adaptation of barley. PMID:27188711

  13. Localization of the Laevigatum powdery mildew resistance gene to barley chromosome 2 by the use of RFLP markers

    DEFF Research Database (Denmark)

    Giese, H.; Holm-Jensen, A.G.; Jensen, H.P.;

    1993-01-01

    The powdery mildew disease resistance gene Ml(La) was found to belong to a locus on barely chromosome 2. We suggest that this locus be designated MlLa. Linkage analysis was carried out on 72 chromosome-doubled, spring-type progeny lines from a cross between the winter var 'Vogelsanger Gold' and t......' and the spring var 'Alf'. A map of chromosome 2 spanning 119 cM and flanked by two peroxidase gene loci was constructed. In addition to the Laevigatum resistance locus the map includes nine RFLP markers, the two peroxidase gene loci and the six-row locus in barley....

  14. The effect of water deficit on the activity of hydrogen peroxide-scavenging enzymes in two barley genotypes

    Directory of Open Access Journals (Sweden)

    Hanna Bandurska

    2014-01-01

    Full Text Available Two barley (Hordeum vulgare L. genotypes, the cv. Aramir and line R567, were subjected to water deficit by immersing their root systems in polyethylene glycol solution of osmotic potential -1.0 MPa. The stress caused a decline in the leaf-relative-water content (RWC and affected membrane damage in both the genotypes. A higher decline in RWC and a higher membrane injury index was observed in R567 in comparison to 'Aramir'. Water deficit induced an increase in the activity of guaiacol peroxidase (GPO and catalse (CAT. A higher increase of CAT than GPO peroxidase activity has been noted in both the genotypes. The results. together with our earlier reports (Bandurska et al. 1997 show that detoxification of hydrogen peroxide under water stress conditions in those two barley genotypes was associated with the action of GPO and CAT, and that the latter was more involved in that process.

  15. Substrate oxidation sites in versatile peroxidase and other basidiomycete peroxidases.

    Science.gov (United States)

    Ruiz-Dueñas, Francisco J; Morales, María; García, Eva; Miki, Yuta; Martínez, María Jesús; Martínez, Angel T

    2009-01-01

    Versatile peroxidase (VP) is defined by its capabilities to oxidize the typical substrates of other basidiomycete peroxidases: (i) Mn(2+), the manganese peroxidase (MnP) substrate (Mn(3+) being able to oxidize phenols and initiate lipid peroxidation reactions); (ii) veratryl alcohol (VA), the typical lignin peroxidase (LiP) substrate; and (iii) simple phenols, which are the substrates of Coprinopsis cinerea peroxidase (CIP). Crystallographic, spectroscopic, directed mutagenesis, and kinetic studies showed that these 'hybrid' properties are due to the coexistence in a single protein of different catalytic sites reminiscent of those present in the other basidiomycete peroxidase families. Crystal structures of wild and recombinant VP, and kinetics of mutated variants, revealed certain differences in its Mn-oxidation site compared with MnP. These result in efficient Mn(2+) oxidation in the presence of only two of the three acidic residues forming its binding site. On the other hand, a solvent-exposed tryptophan is the catalytically-active residue in VA oxidation, initiating an electron transfer pathway to haem (two other putative pathways were discarded by mutagenesis). Formation of a tryptophanyl radical after VP activation by peroxide was detected using electron paramagnetic resonance. This was the first time that a protein radical was directly demonstrated in a ligninolytic peroxidase. In contrast with LiP, the VP catalytic tryptophan is not beta-hydroxylated under hydrogen peroxide excess. It was also shown that the tryptophan environment affected catalysis, its modification introducing some LiP properties in VP. Moreover, some phenols and dyes are oxidized by VP at the edge of the main haem access channel, as found in CIP. Finally, the biotechnological interest of VP is discussed.

  16. RECOMBINANT HORSERADISH PEROXIDASE FOR ANALYTICAL APPLICATIONS

    OpenAIRE

    2013-01-01

    The article deals with prospects of using recombinant horseradish peroxidase in analytical biochemistry and biotechnology. Problems of recombinant horseradish peroxidase cloning in different expression systems, possible approaches to their solution, advantages of recombinant recombinant horseradish peroxidase and recombinant horseradish peroxidase-fusion proteins for immunoassays are considered. Possibility for development of mediatorless bienzyme biosensor for peroxide and metabolites, yield...

  17. Engineering a horseradish peroxidase C stable to radical attacks by mutating multiple radical coupling sites.

    Science.gov (United States)

    Kim, Su Jin; Joo, Jeong Chan; Song, Bong Keun; Yoo, Young Je; Kim, Yong Hwan

    2015-04-01

    Peroxidases have great potential as industrial biocatalysts. In particular, the oxidative polymerization of phenolic compounds catalyzed by peroxidases has been extensively examined because of the advantage of this method over other conventional chemical methods. However, the industrial application of peroxidases is often limited because of their rapid inactivation by phenoxyl radicals during oxidative polymerization. In this work, we report a novel protein engineering approach to improve the radical stability of horseradish peroxidase isozyme C (HRPC). Phenylalanine residues that are vulnerable to modification by the phenoxyl radicals were identified using mass spectrometry analysis. UV-Vis and CD spectra showed that radical coupling did not change the secondary structure or the active site of HRPC. Four phenylalanine (Phe) residues (F68, F142, F143, and F179) were each mutated to alanine residues to generate single mutants to examine the role of these sites in radical coupling. Despite marginal improvement of radical stability, each single mutant still exhibited rapid radical inactivation. To further reduce inactivation by radical coupling, the four substitution mutations were combined in F68A/F142A/F143A/F179A. This mutant demonstrated dramatic enhancement of radical stability by retaining 41% of its initial activity compared to the wild-type, which was completely inactivated. Structure and sequence alignment revealed that radical-vulnerable Phe residues of HPRC are conserved in homologous peroxidases, which showed the same rapid inactivation tendency as HRPC. Based on our site-directed mutagenesis and biochemical characterization, we have shown that engineering radical-vulnerable residues to eliminate multiple radical coupling can be a good strategy to improve the stability of peroxidases against radical attack.

  18. Catalysis of acetoin formation by brewers' yeast pyruvate decarboxylase isozymes.

    Science.gov (United States)

    Stivers, J T; Washabaugh, M W

    1993-12-14

    Catalysis of C(alpha)-proton transfer from 2-(1-hydroxyethyl)thiamin diphosphate (HETDP) by pyruvate decarboxylase isozymes (PDC; EC 4.1.1.1) from Saccharomyces carlsbergensis was investigated by determining the steady-state kinetics of the reaction of [1-L]acetaldehyde (L = H, D, or T) to form acetoin and the primary kinetic isotope effects on the reaction. The PDC isozyme mixture and alpha 4 isozyme (alpha 4-PDC) have different steady-state kinetic parameters and isotope effects for acetoin formation in the presence and absence of the nonsubstrate allosteric effector pyruvamide: pyruvamide activation occurs by stabilization of the acetaldehyde/PDC ternary complex. The magnitudes of primary L(V/K)-type (L = D or T) isotope effects on C(alpha)-proton transfer from alpha 4-PDC-bound HETDP provide no evidence for significant breakdown of the Swain-Schaad relationship that would indicate partitioning of the putative C(alpha)-carbanion/enamine intermediate between HETDP and products. The substrate concentration dependence of the deuterium primary kinetic isotope effects provides evidence for an intrinsic isotope effect of 4.1 for C(alpha)-proton transfer from alpha 4-PDC-bound HETDP. A 1.10 +/- 0.02-fold 14C isotope discrimination against [1,2-14C]acetaldehyde in acetoin formation is inconsistent with a stepwise mechanism, in which the addition step occurs after rate-limiting formation of the C(alpha)-carbanion/enamine as a discrete enzyme-bound intermediate, and provides evidence for a concerted reaction mechanism with an important component of carbon-carbon bond formation in the transition state.

  19. Proteolytic susceptibility of creatine kinase isozymes and arginine kinase.

    Science.gov (United States)

    Ercan, Altan; Grossman, Steven H

    2003-07-11

    The time course and dose-response to proteolysis of three dimeric isozymes of creatine kinase, CK-MM (muscle), CK-BB (brain), and CK-MB (heart) and the homologous monomer, arginine kinase were compared. Chymotrypsin and trypsin cause a rapid and significant loss of intact CK-BB, but limited hydrolysis of CK-MM. After 1h of hydrolysis by chymotrypsin, 80% of CK-MM is intact as judged by quantification of monomers after electrophoresis in sodium dodecyl sulfate. While 50% of the intact monomers of CK-MB remain under these conditions, no CK-BB monomers are detected. These results indicate that treatment with chymotrypsin leads to a CK-MB devoid of the B-subunit. When treated with trypsin for 1h, CK-MM is totally resistant to hydrolysis and all CK-BB is highly degraded. However, CK-MB exhibits approximately 90% intact monomers, indicating survival of intact B-subunit in CK-MB. This suggests that heterodimerization of a B-subunit with an M-subunit may have a protective effect against hydrolysis by trypsin. In view of the considerably larger number of potentially tryptic sensitive sites on the muscle isozyme, the resistance of CK-MM and susceptibility of CK-BB dimers to trypsin implies that differences in subunit tertiary structure are a factor in proteolysis of the homodimeric isozymes. Arginine kinase is rapidly degraded by trypsin, but is minimally affected by chymotrypsin. The finding that both a monomeric (arginine kinase) and dimeric (CK-BB) phosphagen kinase are highly susceptible to proteolysis by trypsin indicates that quaternary structure is not, in and of itself, an advantage in resistance to proteolysis. Since both arginine kinase and muscle creatine kinase are resistant to chymotryptic hydrolysis, it seems unlikely that in general, the increased packing density, which may result from dimerization can account for the stability of CK-MM towards trypsin.

  20. V. PRELIMINARY STUDY ON ISOZYMES OF SHOREA JAVANICA

    Directory of Open Access Journals (Sweden)

    U. JUNIARTI and M. I. J. UMBOH

    1987-01-01

    Full Text Available The detection of genetic variability in natural or man-made populations/ plantations is useful in both basic and applied biology. In addition to the various facets of studies on Shorea javanica already initiated by Torquebiau (1984 and alongside with his recommendations on focus for future research, a study on the genetic aspects of the species should be given important considerations. As the trees are tapped for resin, an important forest product, the genetic basis of the production as well as the range of variation in amount of resin production among t he trees must be known. Coupled with this is a thorough investigation on the differences in pest resistance/susceptability among the trees and their genetic basis. While the assumption (Torquebiau 1984 that trees in natural forest areas are-rarely attacked by diseases because of mycorrhizal fungi is interesting, its confirmation is necessary. If this is true, problems would arise when plants are introduced into a new plantation site as experienced by the Forest Research Institute (Ardikoesuma 1954. Thus, we need to look for pest resistant plants i.e. those that can remain healthy even in the absence of mycorrhizae. The above studies on possible genetic variation could give vital information for development of forest plantations of the species and for breeding and tree improvement strategies. By knowing the extent of genetic variation in natural population or in plantations one could be guided to maintain or increase the genetic base in these areas. Biochemical characters such as isozyme banding patterns have been useful in several areas of plant biology, population genetics, evolution and breeding. Isozymes are detected by starch gel electrophoresis and when their genetic control is established, they could be genetic markers in analyzing variation in morphological or physiological characters. The present study is an attempt to detect the isozymes in leaves, seeds and cotyledons of Shorea

  1. Isozyme analysis of Taenia solium isolates from Mexico and Colombia

    Directory of Open Access Journals (Sweden)

    Pablo Maravilla

    2003-12-01

    Full Text Available Mexican and Colombian Taenia solium cysticerci and some species of Taenia adults were assayed using cellulose acetate electrophoresis to distinguish between isolates. Isozyme patterns for ARK, GOT, G3PD, GPI, and MPI were identical in all cysticerci suggesting homozygotic profiles. G6PD and MDH showed different patterns between Mexican and Colombian cysticerci, suggesting regional differences. ME activity was mainly detected in the adult stage suggesting that this enzyme is active in anaerobic environment, while MDH, detected in cysticerci, could be related to an environment that contains oxygen. Finally, the species of taeniid adults analyzed showed different patterns among them.

  2. Isozyme analysis of Taenia solium isolates from Mexico and Colombia.

    Science.gov (United States)

    Maravilla, Pablo; Valera, Aldo; Souza, Valeria; Martinez-Gordillo, Mario; Flisser, Ana

    2003-12-01

    Mexican and Colombian Taenia solium cysticerci and some species of Taenia adults were assayed using cellulose acetate electrophoresis to distinguish between isolates. Isozyme patterns for ARK, GOT, G3PD, GPI, and MPI were identical in all cysticerci suggesting homozygotic profiles. G6PD and MDH showed different patterns between Mexican and Colombian cysticerci, suggesting regional differences. ME activity was mainly detected in the adult stage suggesting that this enzyme is active in anaerobic environment, while MDH, detected in cysticerci, could be related to an environment that contains oxygen. Finally, the species of taeniid adults analyzed showed different patterns among them.

  3. Correlation of isozyme profiles with genomic sequences of Phytophthora ramorum and its P. sojae orthologues

    NARCIS (Netherlands)

    Man in 't Veld, W.A.; Govers, F.; Meijer, H.J.G.

    2007-01-01

    A correct interpretation of isozyme patterns can be seriously hampered by the lack of supporting genetic data. The availability of the complete genome sequence of Phytophthora ramorum, enabled us to correlate isozyme profiles with the gene models predicted for these enzymes. Thirty-nine P. ramorum s

  4. Structure and function of adenylate kinase isozymes in normal humans and muscular dystrophy patients.

    Science.gov (United States)

    Hamada, M; Takenaka, H; Fukumoto, K; Fukamachi, S; Yamaguchi, T; Sumida, M; Shiosaka, T; Kurokawa, Y; Okuda, H; Kuby, S A

    1987-01-01

    Two isozymes of adenylate kinase from human Duchenne muscular dystrophy serum, one of which was an aberrant form specific to DMD patients, were separated by Blue Sepharose CL-6B affinity chromatography. The separated aberrant form possessed a molecular weight of 98,000 +/- 1,500, whereas the normal serum isozyme had a weight of 87,000 +/- 1,600, as determined by SDS-polyacrylamide gel electrophoresis, gel filtration, and sedimentation equilibrium. The sedimentation coefficients were 5.8 S and 5.6 S for the aberrant form and the normal form, respectively. Both serum isozymes are tetramers. The subunit size of the aberrant isozyme (Mr = 24,700) was very similar to that of the normal human liver isozyme, and the subunit size of the normal isozyme (Mr = 21,700) was very similar to that of the normal human muscle enzyme. The amino acid composition of the normal serum isozyme was similar to that of the muscle-type enzyme, and that of the aberrant isozyme was similar to that of the liver enzyme, with some exceptions in both cases.

  5. Analysis of Genetic diversity and reltionships in local Tunisian barley ...

    African Journals Online (AJOL)

    Yomi

    Key words: Barley, RAPD markers, SSR markers, genetic diversity. INTRODUCTION. Barley ... surveyed by each kind of marker, their distribution ..... that belong to the Center. ..... tagged-site facilitated PCR for barley genome mapping. Theor.

  6. Comparative biochemical characterization of peroxidases (class III) tightly bound to the maize root cell walls and modulation of the enzyme properties as a result of covalent binding.

    Science.gov (United States)

    Hadži-Tašković Šukalović, Vesna; Vuletić, Mirjana; Marković, Ksenija; Cvetić Antić, Tijana; Vučinić, Željko

    2015-01-01

    Comparative biochemical characterization of class III peroxidase activity tightly bound to the cell walls of maize roots was performed. Ionically bound proteins were solubilized from isolated walls by salt washing, and the remaining covalently bound peroxidases were released, either by enzymatic digestion or by a novel alkaline extraction procedure that released covalently bound alkali-resistant peroxidase enzyme. Solubilized fractions, as well as the salt-washed cell wall fragments containing covalently bound proteins, were analyzed for peroxidase activity. Peroxidative and oxidative activities indicated that peroxidase enzymes were predominately associated with walls by ionic interactions, and this fraction differs from the covalently bound one according to molecular weight, isozyme patterns, and biochemical parameters. The effect of covalent binding was evaluated by comparison of the catalytic properties of the enzyme bound to the salt-washed cell wall fragments with the corresponding solubilized and released enzyme. Higher thermal stability, improved resistance to KCN, increased susceptibility to H2O2, stimulated capacity of wall-bound enzyme to oxidize indole-3-acetic acid (IAA) as well as the difference in kinetic parameters between free and bound enzymes point to conformational changes due to covalent binding. Differences in biochemical properties of ionically and covalently bound peroxidases, as well as the modulation of the enzyme properties as a result of covalent binding to the walls, indicate that these two fractions of apoplastic peroxidases play different roles.

  7. Isozymes of bovine intestinal alkaline phosphatase. Characterization and functional studies

    Energy Technology Data Exchange (ETDEWEB)

    Besman, M.J.A.

    1986-01-01

    The membrane-associated alkaline phosphatases of calf and adult bovine small intestines have been isolated to homogeneity by a novel method developed to purify large quantities of enzyme. Chromatofocusing revealed the existence of two isozymes in calf tissue while only one form was present in the adult. The three amphiphilic metallo protein dimers were characterized as to total amino acid and carbohydrate content, zinc stoichiometries and mode of carbohydrate linkage. The molecular relationship between the three enzymes was defined by tryptic peptide HPLC-mapping and N-terminal sequencing, and demonstrated the existence of two calf isozymes of unique primary sequence, only one of which is expressed in the adult animal. In the presence of protease inhibitors, two new, higher M/sub r/ species (66,000 and 62,000 daltons vs 60,000 daltons) of adult bovine alkaline phosphatase were demonstrated by electrophoresis of /sup 32/P/sub i/-labeled tissue, probing gels by autoradiography and Western blotting. The in vivo enzyme was isolated using a modified, rapid procedure; the two higher M/sub r/ species copurified.

  8. Roles of Hydroxynitrile Glucosides in Barley

    DEFF Research Database (Denmark)

    Roelsgaard, Pernille Sølvhøj

    , the defense capability of these compounds requires the activity of a specific β-glucosidase, and this β-glucosidase is not found in barley leaf tissue. Therefore, the role of hydroxynitrile glucosides in barley leaves is unclear. In contrast to acting as defense compounds, it has been suggested......) has been reported in the literature. In this thesis, the role of hydroxynitrile glucosides in the interaction between barley and Bgh is investigated. It is shown that the hydroxynitrile glucoside levels increase over time in barley leaves upon Bgh infection. In addition, isolation of fungal hyphae...

  9. Enumeration of fungi in barley

    CSIR Research Space (South Africa)

    Rabie, CJ

    1997-04-01

    Full Text Available et al., 1975), and affect the resultant beer by causing off-flavours and colours and, in some instances, gushing (Haikara, 1983; Vaag, 1985). Under cer- tain circumstances some fungal species and/or their products may... samples Dominant spccics Kernels inl?cctcd,? (?j6) Table I Fungi isolated from baa-ley kernels As high levels of infection in barley are detri- mental to good quality malt and beer. it is impor- tant to quantify fungal...

  10. High-yield production of manganese peroxidase, lignin peroxidase, and versatile peroxidase in Phanerochaete chrysosporium.

    Science.gov (United States)

    Coconi-Linares, Nancy; Magaña-Ortíz, Denis; Guzmán-Ortiz, Doralinda A; Fernández, Francisco; Loske, Achim M; Gómez-Lim, Miguel A

    2014-11-01

    The white-rot fungus Phanerochaete chrysosporium secretes extracellular oxidative enzymes during secondary metabolism, but lacks versatile peroxidase, an enzyme important in ligninolysis and diverse biotechnology processes. In this study, we report the genetic modification of a P. chrysosporium strain capable of co-expressing two endogenous genes constitutively, manganese peroxidase (mnp1) and lignin peroxidase (lipH8), and the codon-optimized vpl2 gene from Pleurotus eryngii. For this purpose, we employed a highly efficient transformation method based on the use of shock waves developed by our group. The expression of recombinant genes was verified by PCR, Southern blot, quantitative real-time PCR (qRT-PCR), and assays of enzymatic activity. The production yield of ligninolytic enzymes was up to four times higher in comparison to previously published reports. These results may represent significant progress toward the stable production of ligninolytic enzymes and the development of an effective fungal strain with promising biotechnological applications.

  11. Expression analysis of polyphenol oxidase isozymes by active staining method and tissue browning of head lettuce (Lactuca sativa L.).

    Science.gov (United States)

    Noda, Takahiro; Iimure, Kazuhiko; Okamoto, Shunsuke; Saito, Akira

    2017-08-01

    Browning of plant tissue is generally considered attributable to enzymatic oxidation by polyphenol oxidase (PPO). Electrophoresis followed by activity staining has been used as an effective procedure to visually detect and isolate isozymes; however, it has not been applied for examination of various PPO isozymes in lettuce. Our study demonstrated that different lettuce PPO isozymes could be detected at different pH in active staining, and multiple isozymes were detected only under alkaline conditions. As a result, we concluded that activity staining with approximately pH 8 enabled to detect various PPO isozymes in lettuce. By expression analysis of the PPO isozymes after wounding, PPO isozymes that correlated with time-course of tissue browning were detected. The wound-induced PPO may play a key role in enzymatic browning.

  12. Proteomic and activity profiles of ascorbate-glutathione cycle enzymes in germinating barley embryo

    DEFF Research Database (Denmark)

    Bønsager, Birgit Christine; Shahpiri, Azar; Finnie, Christine

    2010-01-01

    Enzymes involved in redox control are important during seed germination and seedling growth. Ascorbate-glutathione cycle enzymes in barley embryo extracts were monitored both by 2D-gel electrophoresis and activity measurements from 4 to 144 h post imbibition (PI). Strikingly different activity...... profiles were observed. No ascorbate peroxidase (APX) activity was present in mature seeds but activity was detected after 24 h PI and increased 14-fold up to 144 h PI. In contrast, dehydroascorbate reductase (DHAR) activity was present at 4 h PI and first decreased by 9-fold until 72 h PI followed by a 5...

  13. Genetics Home Reference: eosinophil peroxidase deficiency

    Science.gov (United States)

    ... invaders. EPX gene mutations reduce or prevent eosinophil peroxidase production or result in a protein that is unstable and nonfunctional. As a result, eosinophils have severely reduced amounts of eosinophil peroxidase or none at all. Other proteins within affected ...

  14. Meloidogyne partityla on Pecan Isozyme Phenotypes and Other Host.

    Science.gov (United States)

    Starr, J L; Tomaszewski, E K; Mundo-Ocampo, M; Baldwin, J G

    1996-12-01

    Meloidogyne sp. from five pecan (Carya illinoensis) orchards in Texas were distinctive in host range and iszoyme profiles from common species of Meloidogyne but were morphologically congruent with Meloidogyne partityla Kleynhans, a species previously known only in South Africa. In addition to pecan, species of walnut (Juglans hindsii and J. regia) and hickory (C. ovata) also were hosts. No reproduction was observed on 15 other plant species from nine families, including several common hosts of other Meloidogyne spp. Three esterase phenotypes and two malate dehydrogenase phenotypes of M. partityla were identified by polyacrylamide gel electrophoresis. Each of these isozyme phenotypes was distinct from those of the more common species M. arenaria, M. hapla, M. incognita, and M. javanica.

  15. Towards isozyme-selective HDAC inhibitors for interrogating disease.

    Science.gov (United States)

    Gupta, Praveer; Reid, Robert C; Iyer, Abishek; Sweet, Matthew J; Fairlie, David P

    2012-01-01

    Histone deacetylase (HDAC) enzymes have emerged as promising targets for the treatment of a wide range of human diseases, including cancers, inflammatory and metabolic disorders, immunological, cardiovascular, and infectious diseases. At present, such applications are limited by the lack of selective inhibitors available for each of the eighteen HDAC enzymes, with most currently available HDAC inhibitors having broad-spectrum activity against multiple HDAC enzymes. Such broad-spectrum activity maybe useful in treating some diseases like cancers, but can be detrimental due to cytotoxic side effects that accompany prolonged treatment of chronic diseased states. Here we summarize progress towards the design and discovery of HDAC inhibitors that are selective for some of the eleven zinc-containing classical HDAC enzymes, and identify opportunities to use such isozyme-selective inhibitors as chemical probes for interrogating the biological roles of individual HDAC enzymes in diseases.

  16. Characterization of cationic acid phosphatase isozyme from rat liver mitochondria.

    Science.gov (United States)

    Fujimoto, S; Murakami, K; Hosoda, T; Yamamoto, Y; Watanabe, K; Morinaka, Y; Ohara, A

    1992-05-01

    Acid phosphatase isozyme was highly purified from rat liver mitochondrial fraction. The enzyme showed an isoelectric point value of above 9.5 on isoelectric focusing, and the apparent molecular weight was estimated to be 32000 by Sephadex G-100 gel filtration or 16000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme catalyzed the hydrolysis of adenosine 5'-triphosphate, adenosine 5'-diphosphate, thiamine pyrophosphate, inorganic pyrophosphate, and phosphoprotein such as casein and phosvitin, but not of several phosphomonoesters, except for p-nitrophenyl phosphate and o-phosphotyrosine. The enzyme was not inhibited by L-(+)-tartrate, and was significantly activated by Fe2+ and reducing agents such as ascorbic acid, L-cysteine,and dithiothreitol. The enzyme was found to be distributed in various rat tissues including liver, spleen, kidney, small intestine, lung, stomach, brain and heart, but not in skeletal muscle.

  17. Issues surrounding health claims for barley.

    Science.gov (United States)

    Ames, Nancy P; Rhymer, Camille R

    2008-06-01

    Government-approved health claims support dietary intervention as a safe and practical approach to improving consumer health and provide industry with regulatory guidelines for food product labels. Claims already allowed in the United States, United Kingdom, Sweden, and The Netherlands for reducing cholesterol through consumption of oat or barley soluble fiber provide a basis for review, but each country may have different criteria for assessing clinical evidence for a physiological effect. For example, the FDA-approved barley health claim was based on a petition that included 39 animal model studies and 11 human clinical trials. Since then, more studies have been published, but with few exceptions, clinical data continue to demonstrate that the consumption of barley products is effective for lowering total and LDL cholesterol. More research is needed to fully understand the mechanism of cholesterol reduction and the role of beta-glucan molecular weight, viscosity, and solubility. In an assessment of the physiological efficacy of a dietary intervention, consideration should also be given to the potential impact of physical and thermal food-processing treatments and genotypic variation in the barley source. New barley cultivars have been generated specifically for food use, possessing increased beta-glucan, desirable starch composition profiles, and improved milling/processing traits. These advances in barley production, coupled with the establishment of a government-regulated health claim for barley beta-glucan, will stimulate new processing opportunities for barley foods and provide consumers with reliable, healthy food choices.

  18. Molecular characterization of two lipoxygenases from barley

    NARCIS (Netherlands)

    Mechelen, J.R. van; Schuurink, R.C.; Smits, M.; Graner, A.; Douma, A.C.; Sedee, N.J.A.; Schmitt, N.F.; Valk, B.E.

    1999-01-01

    Two full-length lipoxygenase cDNA sequences (LoxB and LoxC) from barley (Hordeum distichum cv. L. Triumph) are described. The cDNAs share high homology with the barley LoxA cDNA. Southern blotting experiments indicate single copy numbers of the three lipoxygenase genes. RFLP mapping revealed the pre

  19. Rachiplusia nu larva as a biofactory to achieve high level expression of horseradish peroxidase.

    Science.gov (United States)

    Romero, Lucía Virginia; Targovnik, Alexandra Marisa; Wolman, Federico Javier; Cascone, Osvaldo; Miranda, María Victoria

    2011-05-01

    A process based on orally-infected Rachiplusia nu larvae as biological factories for expression and one-step purification of horseradish peroxidase isozyme C (HRP-C) is described. The process allows obtaining high levels of pure HRP-C by membrane chromatography purification. The introduction of the partial polyhedrin homology sequence element in the target gene increased HRP-C expression level by 2.8-fold whereas it increased 1.8-fold when the larvae were reared at 27 °C instead of at 24 °C, summing up a 4.6-fold overall increase in the expression level. Additionally, HRP-C purification by membrane chromatography at a high flow rate greatly increase D the productivity without affecting the resolution. The V(max) and K(m) values of the recombinant HRP-C were similar to those of the HRP from Armoracia rusticana roots.

  20. Calcium homeostasis in barley aleurone

    Energy Technology Data Exchange (ETDEWEB)

    Jones, R.L.

    1990-02-21

    Under the auspices of the Department of Energy we investigated calcium homeostasis in aleurone cells of barley. This investigation was initiated to explore the role played by extracellular Ca{sup 2+} in gibberellic acid (GA)-induced synthesis and secretion of hydrolases in the aleurone layer. We have focused our attention on four topics that relate to the role of Ca{sup 2+} in regulating the synthesis of {alpha}-amylase. First, we determined the stoichiometry of Ca{sup 2+} binding to the two principal classes of barley {alpha}-amylase and examined some of the biochemical and physical properties of the native and Ca{sup 2+}-depleted forms of the enzyme. Second, since {alpha}-amylase is a Ca{sup 2+} containing metalloenzyme that binds one atom of Ca{sup 2+} per molecule, we developed methods to determine the concentration of Ca{sup 2+} in the cytosol of the aleurone cell. We developed a technique for introducing Ca{sup 2+}-sensitive dyes into aleurone protoplasts that allows the measurement of Ca{sup 2+} in both cytosol and endoplasmic reticulum (ER). Third, because the results of our Ca{sup 2+} measurements showed higher levels of Ca{sup 2+} in the ER than in the cytosol, we examined Ca{sup 2+} transport into the ER of control and GA-treated aleurone tissue. And fourth, we applied the technique of patch-clamping to the barley aleurone protoplast to examine ion transport at the plasma membrane. Our results with the patch-clamp technique established the presence of K{sup +} channels in the plasma membrane of the aleurone protoplast, and they showed that this cell is ideally suited for the application of this methodology for studying ion transport. 34 refs.

  1. Akt3 is a privileged first responder in isozyme-specific electrophile response.

    Science.gov (United States)

    Long, Marcus J C; Parvez, Saba; Zhao, Yi; Surya, Sanjna L; Wang, Yiran; Zhang, Sheng; Aye, Yimon

    2017-03-01

    Isozyme-specific post-translational regulation fine tunes signaling events. However, redundancy in sequence or activity renders links between isozyme-specific modifications and downstream functions uncertain. Methods to study this phenomenon are underdeveloped. Here we use a redox-targeting screen to reveal that Akt3 is a first-responding isozyme sensing native electrophilic lipids. Electrophile modification of Akt3 modulated downstream pathway responses in cells and Danio rerio (zebrafish) and markedly differed from Akt2-specific oxidative regulation. Digest MS sequencing identified Akt3 C119 as the privileged cysteine that senses 4-hydroxynonenal. A C119S Akt3 mutant was hypomorphic for all downstream phenotypes shown by wild-type Akt3. This study documents isozyme-specific and chemical redox signal-personalized physiological responses.

  2. Resistance to Barley Leaf Stripe

    DEFF Research Database (Denmark)

    Nørgaard Knudsen, J. C.

    1986-01-01

    in well adapted Northwest European spring cultivars. Virulence matching two hitherto not overcome resistances was demonstrated. Differences in apparent race nonspecific or partial resistance were also present, changing the percentage of infected plants of susceptible genotypes from about 20 to 44 per cent.......Ten barley [Hordeum vulgare] genotypes were inoculated with twelve isolates of Pyrenophora graminea of diverse European and North African origin. Race specific resistance occurred. Four, possibly five, genetically different sources of race-specific resistance were found, three of them occurring...

  3. Recombinant horseradish peroxidase: production and analytical applications.

    Science.gov (United States)

    Grigorenko, V G; Andreeva, I P; Rubtsova, M Yu; Egorov, A M

    2015-04-01

    Horseradish peroxidase is a key enzyme in bio- and immunochemical analysis. New approaches in functional expression of the peroxidase gene in E. coli cells and the subsequent refolding of the resulting protein yield a recombinant enzyme that is comparable in its spectral and catalytic characteristics to the native plant peroxidase. Genetic engineering approaches allow production of recombinant peroxidase conjugates with both protein antigens and Fab antibody fragments. The present article reviews the use of recombinant horseradish peroxidase as the marker enzyme in ELISA procedures as well as in amperometric sensors based on direct electron transfer.

  4. RECOMBINANT HORSERADISH PEROXIDASE FOR ANALYTICAL APPLICATIONS

    Directory of Open Access Journals (Sweden)

    А.M. Egorov

    2012-08-01

    Full Text Available The article deals with prospects of using recombinant horseradish peroxidase in analytical biochemistry and biotechnology. Problems of recombinant horseradish peroxidase cloning in different expression systems, possible approaches to their solution, advantages of recombinant recombinant horseradish peroxidase and recombinant horseradish peroxidase-fusion proteins for immunoassays are considered. Possibility for development of mediatorless bienzyme biosensor for peroxide and metabolites, yielding hydrogen peroxide during their transformations, based on co-adsorption of recombinant horseradish peroxidase and the appropriate oxidase was demonstrated. The possibility to produce a fully active recombinant conjugate of recombinant horseradish peroxidase with human heart-type fatty acid binding protein, which may be used in competitive immunoassay for clinical diagnosis of acute myocardial infarction, and recombinant conjugates (N- and C-terminus of recombinant horseradish peroxidase with Fab-fragments of the antibody against atrazine, which may be applied for atrazine pesticides detection, are demonstra ted for the first time.

  5. First comparative phenetic studies of Argentinean species of Acacia (Fabaceae), using morphometric, isozymal, and RAPD approaches.

    Science.gov (United States)

    Casiva, Paola V; Saidman, Beatriz O; Vilardi, Juan C; Cialdella, Ana M

    2002-05-01

    Morphological and genetic diversity among Acacia aroma, A. macracantha, A. caven, and A. furcatispina were studied with morphometric, isozymal, and RAPD approaches. The analysis of seven isozyme systems revealed 21 loci, and RAPD analysis showed 34 loci. Most of these loci allowed us to differentiate the species, with the exception of A. aroma and A. macracantha, the two most similar species. The levels of genetic variability estimated by isozymes were higher than those obtained from RAPD analyses. Morphometric characters showed highly significant differences among the species, although A. aroma and A. macracantha are differentiated only by thorn length. The phenogram obtained from isozyme data is consistent with morphological data. The RAPD phenogram based on allelic frequencies showed agreement with morphological and isozymal approaches only at the intraspecific levels, while the RAPD phenogram based on Nei and Li's similarity measures agreed with the phenograms constructed from isozyme and morphological data. High similarities and high indirect gene flow were found between A. aroma and A. macracantha, results that call the relationship between them into question.

  6. HIV-1 production is specifically associated with human NMT1 long form in human NMT isozymes.

    Science.gov (United States)

    Takamune, Nobutoki; Gota, Kayoko; Misumi, Shogo; Tanaka, Kenzo; Okinaka, Shigetaka; Shoji, Shozo

    2008-02-01

    The N-myristoylation of the N-terminal of human immunodeficiency virus type-1 (HIV-1) Pr55(gag) by human N-myristoyltransferase (hNMT) is a prerequisite modification for HIV-1 production. hNMT consists of multiple isozymes encoded by hNMT1 and hNMT2. The hNMT1 isozyme consists of long, medium, and short forms. Here, we investigated which isozyme is crucial for HIV-1 production. Human embryonic kidney (HEK) 293 cells transfected with infectious HIV-1 vectors were used as models of HIV-1-infected cells in this study. The significant reduction in HIV-1 production and the failure of the specific localization of Pr55(gag) in a detergent-resistant membrane fraction were dependent on the knockdown of the different forms of the hNMT1 isozyme but not of the hNMT2 isozyme. Additionally, the coexpression of an inactive mutant hNMT1 isozyme, namely the hNMT1 long form (hNMT1(L)), but not that of other hNMT mutants resulted in a significant reduction in HIV-1 production. These results strongly suggest that HIV-1 production is specifically associated with hNMT1, particularly hNMT1(L), but not with hNMT2 in vivo, contributing to the understanding of a step in HIV-1 replication.

  7. Isozymes of beta-N-Acetylhexosaminidase from Pea Seeds (Pisum sativum L.).

    Science.gov (United States)

    Harley, S M; Beevers, L

    1987-12-01

    Four isozymes of beta-N-acetylhexosaminidase (beta-NAHA) from pea seeds (Pisum sativum L.) have been separated, with one, designated beta-NAHA-II, purified to apparent homogeneity by means of an affinity column constructed by ligating p-aminophenyl-N-acetyl-beta-d-thioglucosaminide to Affi-Gel 202. The other three isozymes have been separated and purified 500- to 1750-fold by chromatography on Concanavalin A-Sepharose, Zn(2+) charged immobilized metal affinity chromatography, hydrophobic chromatography, and ion exchange chromatography on CM-Sephadex. All four isozymes are located in the protein bodies of the cotyledons. The molecular weight of each isozyme is 210,000. beta-NAHA-II is composed of two heterogenous subunits. The subunits are not held together by disulfide bonds, but sulfhydryl groups are important for catalysis. All four isozymes release p-nitrophenol from both p-nitrophenyl-N-acetyl-beta-d-glucosaminide and p-nitrophenyl-N-acetyl-beta-d-galactosaminide. The ratio of activity for hydrolysis of the two substrates is pH dependent. The K(m) value for the two substrates and pH optima of the isozymes are comparable to beta-NAHAs from other plant sources.

  8. Cisgenic barley for animal feed

    DEFF Research Database (Denmark)

    Holme, Inger; Dionisio, Giuseppe; Brinch-Pedersen, Henrik

    2011-01-01

    for Cisgenesis. Recently, Dionisio et al. (2011) have cloned and characterized phytases belonging to the purple acid phosphatases (PAPs) in barley. We have isolated the genomic PAP-clone of the isoform expressed during grain filling including 2.3 kb of the promoter region and 600 bp of the terminator region...... using a genomic barley lambda library. The clone has been inserted into a Cisgenic Agrobacterium vector where both the gene of interest and the selection gene are flanked by their own T-DNA borders in order to promote integration of the two genes at unlinked places in the plant genome. T0-plants show...... increases in the phytase activity of mature seeds from 1350 in wild type to 7500 FTU/kg in T0-plants. We have identified two Cisgenic T1-lines without selection gene and vector backbone but with one additional genomic clone of the phytase gene. Lines homozygous for the additional cisgene show 2-3 fold...

  9. Control of. cap alpha. -amylase mRNA accumulation by gibberellic acid and calcium in barley aleurone layers

    Energy Technology Data Exchange (ETDEWEB)

    Deikman, J.; Jones, R.L.

    1985-01-01

    Pulse-labeling of barley (Hordeum vulgare L. cv Himalaya) aleurone layers incubated for 13 hours in 2.5 micromolar gibberellic acid (GA/sub 3/) with or without 5 millimolar CaCl/sub 2/ shows that ..cap alpha..-amylase isozymes 3 and 4 are not synthesized in vivo in the absence of Ca/sup 2 +/. No difference was observed in ..cap alpha..-amylase mRNA levels between layers incubated for 12 hours in 2.5 micromolar GA/sub 3/ with 5 millimolar CaCl/sub 2/ and layers incubated in GA/sub 3/ alone. RNA isolated from layers incubated for 12 hours in GA/sub 3/ with and without CA/sup 2 +/. A cDNA clone for ..cap alpha..-amylase was isolated and used to measure ..cap alpha..-amylase mRNA levels in aleurone layers incubated in the presence and absence of Ca/sup 2 +/ was translated in vitro and was found to produce the same complement of translation products regardless of the presence of Ca/sup 2 +/ in the incubation medium. Immunoprecipitation of translation products showed that the RNA for ..cap alpha..-amylase synthesized in Ca/sup 2 +/-deprived aleurone layers was translatable. Ca/sup 2 +/ is required for the synthesis of ..cap alpha..-amylase isozymes 3 and 4 at a step after mRNA accumulation and processing.

  10. Stable disruption of ethanol production by deletion of the genes encoding alcohol dehydrogenase isozymes in Saccharomyces cerevisiae.

    Science.gov (United States)

    Ida, Yoshihiro; Furusawa, Chikara; Hirasawa, Takashi; Shimizu, Hiroshi

    2012-02-01

    We analyzed the effects of the deletions of genes encoding alcohol dehydrogenase (ADH) isozymes of Saccharomyces cerevisiae. The decrease in ethanol production by ADH1 deletion alone could be partially compensated by the upregulation of other isozyme genes, while the deletion of all known ADH isozyme genes stably disrupted ethanol production. Copyright © 2011 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  11. Carbonic anhydrase isozymes Ⅸ and Ⅻ in gastric tumors

    Institute of Scientific and Technical Information of China (English)

    Mari Leppilampi; Juha Saarnio; Tuomo J. Karttunen; Jyrki Kivel(a); Silvia Pastorekov(a); Jaromir Pastorek; Abdul Waheed; William S. Sly; Seppo Parkkila

    2003-01-01

    AIM: To systematically study the expression of carbonic anhydrase (CA) isowmes Ⅸ and Ⅻ in gastric tumors.METHODS: We analyzed a representative series of specimens from non-neoplastic gastric mucosa and from various dysplastic and neoplastic gastric lesions for the expression of CA IX and XII. Immunohistochemical staining was performed using isozyme-specific antibodies and biotinstreptavidin complex method.RESULTS: CA IX was highly expressed in the normal gastric mucosa and remained positive in many gastric tumors. In adenomas, CA IX expression significantly decreased towards the high grade dysplasia. However, the expression resumed back to the normal level in well differentiated adenocarcinomas,while it again declined in carcinomas with less differentiation.In comparison, CA Ⅻ showed no or weak immunoreaction in the normal gastric mucosa and was slightly increased in tumors.CONCLUSION: These results demonstrate that CA Ⅸexpression is sustained in several types of gastric tumors.The variations observed in the CA Ⅸ levels support the concept that gastric adenomas and carcinomas are distinct entities and do not represent progressive steps of a single pathway.

  12. A fluorogenic, small molecule reporter for mammalian phospholipase C isozymes.

    Science.gov (United States)

    Huang, Weigang; Hicks, Stephanie N; Sondek, John; Zhang, Qisheng

    2011-03-18

    Phospholipase C isozymes (PLCs) catalyze the conversion of the membrane lipid phosphatidylinositol 4,5-bisphosphate (PIP(2)) into two second messengers, inositol 1,4,5-trisphosphate and diacylglycerol. This family of enzymes are key signaling proteins that regulate the physiological responses of many extracellular stimuli such as hormones, neurotransmitters, and growth factors. Aberrant regulation of PLCs has been implicated in various diseases including cancer and Alzheimer's disease. How, when, and where PLCs are activated under different cellular contexts are still largely unknown. We have developed a fluorogenic PLC reporter, WH-15, that can be cleaved in a cascade reaction to generate fluorescent 6-aminoquinoline. When applied in enzymatic assays with either pure PLCs or cell lysates, this reporter displays more than a 20-fold fluorescence enhancement in response to PLC activity. Under assay conditions, WH-15 has comparable K(m) and V(max) with the endogenous PIP(2). This novel reporter will likely find broad applications that vary from imaging PLC activity in live cells to high-throughput screening of PLC inhibitors.

  13. Accelerated evolution of snake venom phospholipase A2 isozymes for acquisition of diverse physiological functions.

    Science.gov (United States)

    Ogawa, T; Nakashima, K; Nobuhisa, I; Deshimaru, M; Shimohigashi, Y; Fukumaki, Y; Sakaki, Y; Hattori, S; Ohno, M

    1996-01-01

    The nucleotide sequences of two cDNAs and four genes encoding Trimeresurus gramineus venom gland phospholipase A2 (PLA2) isozymes were determined and compared internally and externally with those encoding Trimeresurus flavoviridis venom gland PLA2 isozymes. It was revealed that the protein-coding regions are much more diversified than the 5' and 3' untranslated regions (UTRs) and the introns except for the signal peptide domain. The numbers of nucleotide substitutions per site (KN) for the UTRs and the introns were approximately one-quarter of the numbers of nucleotide substitutions per synonymous site (KS) for the protein-coding regions and were at the same level as the KN value of T. gramineus and T. flavoviridis TATA box-binding protein (TBP) genes, indicating that the protein-coding regions of PLA2 isozyme genes are unusually variable and that the UTRs including the introns of venom gland PLA2 isozyme genes have evolved at similar rate to those of non-venomous genes. The numbers of nucleotide substitutions per non-synonymous site (KA) values were close to or larger than the KS values for the protein-coding regions in venom gland PLA2 isozyme genes, indicating that the protein-coding regions of snake venom gland PLA2 isozyme genes have evolved via accelerated evolution. Furthermore, the evolutionary trees derived from the combined sequences of the 5' and 3' UTRs and the signal peptide domain of cDNAs were in accord with the consequences from taxonomy. In contrast, the evolutionary trees from the mature protein-coding region sequences of cDNAs and from the amino acid sequences showed random patterns. Estimations of nucleotide divergence of genes and the phylogenetic analysis reveal that snake venom group IJ PLA2 isozyme genes have been evolving under adaptive pressure to acquire new physiological activities.

  14. Reciprocal regulation of transcription factors and PLC isozyme gene expression in adult cardiomyocytes.

    Science.gov (United States)

    Singal, Tushi; Dhalla, Naranjan S; Tappia, Paramjit S

    2010-06-01

    By employing a pharmacological approach, we have shown that phospholipase C (PLC) activity is involved in the regulation of gene expression of transcription factors such as c-Fos and c-Jun in cardiomyocytes in response to norepinephrine (NE). However, there is no information available regarding the identity of specific PLC isozymes involved in the regulation of c-Fos and c-Jun or on the involvement of these transcription factors in PLC isozyme gene expression in adult cardiomyocytes. In this study, transfection of cardiomyocytes with PLC isozyme specific siRNA was found to prevent the NE-mediated increases in the corresponding PLC isozyme gene expression, protein content and activity. Unlike PLC gamma(1) gene, silencing of PLC beta(1), beta(3) and delta(1) genes with si RNA prevented the increases in c-Fos and c-Jun gene expression in response to NE. On the other hand, transfection with c-Jun si RNA suppressed the NE-induced increase in c-Jun as well as PLC beta(1), beta(3) and delta(1) gene expression, but had no effect on PLC gamma(1) gene expression. Although transfection of cardiomyocytes with c-Fos si RNA prevented NE-induced expression of c-Fos, PLC beta(1) and PLC beta(3) genes, it did not affect the increases in PLC delta(1) and PLC gamma(1) gene expression. Silencing of either c-Fos or c-Jun also depressed the NE-mediated increases in PLC beta(1), beta(3) and gamma(1) protein content and activity in an isozyme specific manner. Furthermore, silencing of all PLC isozymes as well as of c-Fos and c-Jun resulted in prevention of the NE-mediated increase in atrial natriuretic factor gene expression. These findings, by employing gene silencing techniques, demonstrate that there occurs a reciprocal regulation of transcription factors and specific PLC isozyme gene expression in cardiomyocytes.

  15. Lignin peroxidase functionalities and prospective applications

    OpenAIRE

    Falade, Ayodeji O.; Nwodo, Uchechukwu U.; Iweriebor, Benson C.; Green, Ezekiel; Leonard V. Mabinya; Okoh, Anthony I.

    2016-01-01

    Abstract Ligninolytic extracellular enzymes, including lignin peroxidase, are topical owing to their high redox potential and prospective industrial applications. The prospective applications of lignin peroxidase span through sectors such as biorefinery, textile, energy, bioremediation, cosmetology, and dermatology industries. The litany of potentials attributed to lignin peroxidase is occasioned by its versatility in the degradation of xenobiotics and compounds with both phenolic and non‐phe...

  16. Evidence for peroxidase activity in Caralluma umbellata.

    Science.gov (United States)

    Achar, Raghu Ram; Venkatesh, B K; Sharanappa, P; Priya, B S; Swamy, S Nanjunda

    2014-08-01

    Vast applications of peroxidases create an increasing demand to characterize peroxidases from new sources with more applicability potential. The aim of the present study was to check the presence of peroxidase activity from Caralluma umbellata. This is the first report on the C. umbellata peroxidase (CUP). The presence of peroxidase was revealed by the histochemical analysis of the stem sections, zymographic studies, and in vitro peroxidase activity assay using various reducing substrates viz., 2, 2'-azinobis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), guaiacol, o-dianisidine, and ferulic acid. The band pattern in zymogram confirms that CUP has a molecular weight less than that of horseradish peroxidase (44 kDa). Comparative evaluation of peroxidase activity of CUP with respect to horseradish peroxidase (HRP) indicates that CUP catalyzes ABTS and ferulic acid in a similar pattern as HRP but with guaiacol, the extent of catalysis shown by CUP over HRP is high. The standard inhibitors sodium azide and sodium meta bisulphite inhibited CUP activity in a dose dependent manner.

  17. DyP-type peroxidases comprise a novel heme peroxidase family.

    Science.gov (United States)

    Sugano, Y

    2009-04-01

    Dye-decolorizing peroxidase (DyP) is produced by a basidiomycete (Thanatephorus cucumeris Dec 1) and is a member of a novel heme peroxidase family (DyP-type peroxidase family) that appears to be distinct from general peroxidases. Thus far, 80 putative members of this family have been registered in the PeroxiBase database (http://peroxibase.isbsib.ch/) and more than 400 homologous proteins have been detected via PSI-BLAST search. Although few studies have characterized the function and structure of these proteins, they appear to be bifunctional enzymes with hydrolase or oxygenase, as well as typical peroxidase activities. DyP-type peroxidase family suggests an ancient root compared with other general peroxidases because of their widespread distribution in the living world. In this review, firstly, an outline of the characteristics of DyP from T. cucumeris is presented and then interesting characteristics of the DyP-type peroxidase family are discussed.

  18. Functional Analysis of Barley Powdery Mildew Effector Candidates and Identification of their Barley Targets

    DEFF Research Database (Denmark)

    Ahmed, Ali Abdurehim

    The genome of barley powdery mildew fungus (Blumeria graminis f. sp. hordei, Bgh) encodes around 500 Candidate Secreted Effector Proteins (CSEPs), which are believed to be delivered to the barley cells either to interfere with plant defence and/or promote nutrient uptake. So far, little is known...

  19. 2015 nationwide survey revealed Barley stripe mosaic virus in Korean barley fields

    Science.gov (United States)

    A seed-transmitted virus has consistently caused significant economic damage to barley crops in Korea in recent years, and may be increasing because many farmers save seed for replanting. Because some barley seed is imported, there is the potential for introduction of new seed-transmitted viruses, c...

  20. Accelerated evolution of Trimeresurus okinavensis venom gland phospholipase A2 isozyme-encoding genes.

    Science.gov (United States)

    Nobuhisa, I; Nakashima, K; Deshimaru, M; Ogawa, T; Shimohigashi, Y; Fukumaki, Y; Sakaki, Y; Hattori, S; Kihara, H; Ohno, M

    1996-06-26

    Three Trimeresurus okinavensis (To; himehabu snake, Crotalinae) venom gland phospholipase A2 (PLA2) isozymeencoding genes, gPLA2-o1, gPLA2-o2 and gPLA2-o3, were isolated from its genomic DNA library. The nucleotide (nt) sequence analysis revealed that two of the three genes (gPLA2-o2 and gPLA2-o3) occasionally have been converted to inactivated genes by introduction of one base insertion or substitution. It was confirmed from Southern blot analysis that the To haploid genome contains only three venom gland PLA2 isozyme genes herein isolated. Comparison of these genes showed that nonsynonymous nt substitutions have occurred more frequently than synonymous nt substitutions in the protein-coding regions, except for the signal-peptide coding domain, implying that To venom gland PLA2 isozyme genes have evolved via accelerated evolution. Such an evolutionary feature of To venom gland PLA2 isozyme genes proves the general universality of accelerated evolution previously drawn for venom gland PLA2 isozyme genes of other crotalinae snakes. The variability in the mature protein-coding regions of three To venom gland PLA2 isozyme genes appears to have been brought about by natural selection for point mutations.

  1. CHARACTERIZATION OF LACTATE DEHYDROGENASE ISOZYME PATTERN AND MORPHOLOGY OF THREE MARINE FISH CELL LINES

    Institute of Scientific and Technical Information of China (English)

    郭华荣; 张士璀; 李红岩; 童裳亮; 相建海

    2002-01-01

    Three continuous marine fish cell lines of FG (i. e., Hounder Gill) from flounder (Paralichthys olivaceus) gill, SPH (i. e. , Sea Perch Heart) from sea perch (Lateolabrax japonicus) heart and RSBF (i.e., Red Sea Bream Fin) from red sea bream (Pagrosomus major) fin, were characterized by lactate dehydrogenase (LDH) isozyme and morphological analysis. The LDH isozyme patterns of these three cell lines and their corresponding tissues of origin were investigated and compared. The results showed: (1) No difference was found in the LDH isozyme patterns of FG and flounder gill tissue. However, the LDH isozyme patterns of SPH and RSBF were significantly different from their corresponding tissues of origin; (2) LDH isozyme patterns of FG, SPH and RSBF were markedly different from each other and could serve as genetic markers for species identification and detection of cross contamination. Morphological change analysis of these three cell lines in comparison to their original tissues indicated that FG cells still appeared epithelioid without morphological transformation. However, morphological changes were found in SPH and RSBF compared to their original tissues. Therefore, the cellular morphology was still plastic in the relatively stable culture conditions, and it was possible that change of LDH patterns wasrelated to morphological changes of fish cells in vitro.

  2. Expression of steroid 5α-reductase isozymes in prostate of adult rats after environmental stress.

    Science.gov (United States)

    Sánchez, Pilar; Torres, Jesús M; Castro, Beatriz; Olmo, Asunción; del Moral, Raimundo G; Ortega, Esperanza

    2013-01-01

    The elevated incidence of prostate cancer and benign prostatic hypertrophy is a cause of increasing public health concern in the Western world. The normal and pathological growth of the prostate are both dependent on stimulation by dihydrotestosterone, which is synthesized from circulating testosterone by two 5α-reductase (5α-R) isozymes, 5α-reductase type 1 (5α-R1) and 5α-reductase type 2 (5α-R2). Both isozymes have been implicated in prostate disease. We used quantitative RT-PCR and immunohistochemistry, respectively, to quantify mRNA and protein levels of 5α-R isozymes in the ventral prostate of adult rats under environmental stress conditions analogous to those found in some common workplace situations, i.e. artificial light, excessive heat, and the sensation of immobility in a small space. Transcription and expression levels of both 5α-R isozymes were significantly higher in environmentally stressed rats than in unstressed rats. Increased 5α-R isozyme levels may play a role in the development or maintenance of prostate disease. Further research is warranted to explore these effects of environmental stress on human health and their implications for environmental and occupational health policies. © 2012 The Authors Journal compilation © 2012 FEBS.

  3. AMP deaminase histochemical activity and immunofluorescent isozyme localization in rat skeletal muscle

    Science.gov (United States)

    Thompson, J. L.; Sabina, R. L.; Ogasawara, N.; Riley, D. A.

    1992-01-01

    The cellular distribution of AMP deaminase (AMPda) isozymes was documented for rat soleus and plantaris muscles, utilizing immunofluorescence microscopy and immunoprecipitation methods. AMPda is a ubiquitous enzyme existing as three distinct isozymes, A, B and C, which were initially purified from skeletal muscle, liver (and kidney), and heart, respectively. AMPda-A is primarily concentrated subsarcolemmally and intermyofibrillarly within muscle cells, while isozymes B and C are concentrated within non-myofiber elements of muscle tissue. AMPda-B is principally associated with connective tissues surrounding neural elements and the muscle spindle capsule, and AMPda-C is predominantly associated with circulatory elements, such as arterial and venous walls, capillary endothelium, and red blood cells. These specific localizations, combined with documented differences in kinetic properties, suggest multiple functional roles for the AMPda isozymes or temporal segregation of similar AMPda functions. Linkage of the AMPda substrate with adenosine production pathways at the AMP level and the localization of isozyme-C in vascular tissue suggest a regulatory role in the microcirculation.

  4. Turning points in the evolution of peroxidase-catalase superfamily: molecular phylogeny of hybrid heme peroxidases.

    Science.gov (United States)

    Zámocký, Marcel; Gasselhuber, Bernhard; Furtmüller, Paul G; Obinger, Christian

    2014-12-01

    Heme peroxidases and catalases are key enzymes of hydrogen peroxide metabolism and signaling. Here, the reconstruction of the molecular evolution of the peroxidase-catalase superfamily (annotated in pfam as PF00141) based on experimentally verified as well as numerous newly available genomic sequences is presented. The robust phylogenetic tree of this large enzyme superfamily was obtained from 490 full-length protein sequences. Besides already well-known families of heme b peroxidases arranged in three main structural classes, completely new (hybrid type) peroxidase families are described being located at the border of these classes as well as forming (so far missing) links between them. Hybrid-type A peroxidases represent a minor eukaryotic subfamily from Excavates, Stramenopiles and Rhizaria sharing enzymatic and structural features of ascorbate and cytochrome c peroxidases. Hybrid-type B peroxidases are shown to be spread exclusively among various fungi and evolved in parallel with peroxidases in land plants. In some ascomycetous hybrid-type B peroxidases, the peroxidase domain is fused to a carbohydrate binding (WSC) domain. Both here described hybrid-type peroxidase families represent important turning points in the complex evolution of the whole peroxidase-catalase superfamily. We present and discuss their phylogeny, sequence signatures and putative biological function.

  5. The Hydrogen Sulfide Donor NaHS Delays Programmed Cell Death in Barley Aleurone Layers by Acting as an Antioxidant.

    Science.gov (United States)

    Zhang, Ying-Xin; Hu, Kang-Di; Lv, Kai; Li, Yan-Hong; Hu, Lan-Ying; Zhang, Xi-Qi; Ruan, Long; Liu, Yong-Sheng; Zhang, Hua

    2015-01-01

    H2S is a signaling molecule in plants and animals. Here we investigated the effects of H2S on programmed cell death (PCD) in barley (Hordeum vulgare L.) aleurone layers. The H2S donor NaHS significantly delayed PCD in aleurone layers isolated from imbibed embryoless barley grain. NaHS at 0.25 mM effectively reduced the accumulation of superoxide anion (·O2 (-)), hydrogen peroxide (H2O2), and malondialdehyde (MDA), promoted the activity of superoxide dismutase (SOD), guaiacol peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX), and decreased those of lipoxygenase (LOX) in isolated aleurone layers. Quantitative-PCR showed that NaHS treatment of aleurone tissue led to enhanced transcript levels of the antioxidant genes HvSOD1, HvAPX, HvCAT1, and HvCAT2 and repressed transcript levels of HvLOX (lipoxygenase gene) and of two cysteine protease genes HvEPA and HvCP3-31. NaHS treatment in gibberellic acid- (GA-) treated aleurone layers also delayed the PCD process, reduced the content of ·O2 (-), and increased POD activity while decreasing LOX activity. Furthermore, α-amylase secretion in barley aleurone layers was enhanced by NaHS treatment regardless of the presence or absence of GA. These data imply that H2S acted as an antioxidant in delaying PCD and enhances α-amylase secretion regardless of the presence of GA in barley aleurone layers.

  6. The Hydrogen Sulfide Donor NaHS Delays Programmed Cell Death in Barley Aleurone Layers by Acting as an Antioxidant

    Directory of Open Access Journals (Sweden)

    Ying-Xin Zhang

    2015-01-01

    Full Text Available H2S is a signaling molecule in plants and animals. Here we investigated the effects of H2S on programmed cell death (PCD in barley (Hordeum vulgare L. aleurone layers. The H2S donor NaHS significantly delayed PCD in aleurone layers isolated from imbibed embryoless barley grain. NaHS at 0.25 mM effectively reduced the accumulation of superoxide anion (·O2-, hydrogen peroxide (H2O2, and malondialdehyde (MDA, promoted the activity of superoxide dismutase (SOD, guaiacol peroxidase (POD, catalase (CAT, and ascorbate peroxidase (APX, and decreased those of lipoxygenase (LOX in isolated aleurone layers. Quantitative-PCR showed that NaHS treatment of aleurone tissue led to enhanced transcript levels of the antioxidant genes HvSOD1, HvAPX, HvCAT1, and HvCAT2 and repressed transcript levels of HvLOX (lipoxygenase gene and of two cysteine protease genes HvEPA and HvCP3-31. NaHS treatment in gibberellic acid- (GA- treated aleurone layers also delayed the PCD process, reduced the content of ·O2-, and increased POD activity while decreasing LOX activity. Furthermore, α-amylase secretion in barley aleurone layers was enhanced by NaHS treatment regardless of the presence or absence of GA. These data imply that H2S acted as an antioxidant in delaying PCD and enhances α-amylase secretion regardless of the presence of GA in barley aleurone layers.

  7. Proteome analysis of grain filling and seed maturation in barley.

    Science.gov (United States)

    Finnie, Christine; Melchior, Sabrina; Roepstorff, Peter; Svensson, Birte

    2002-07-01

    In monocotyledonous plants, the process of seed development involves the deposition of reserves in the starchy endosperm and development of the embryo and aleurone layer. The final stages of seed development are accompanied by an increase in desiccation tolerance and drying out of the mature seed. We have used two-dimensional gel electrophoresis for a time-resolved study of the changes in proteins that occur during seed development in barley (Hordeum vulgare). About 1,000 low-salt extractable protein spots could be resolved on the two-dimensional gels. Protein spots were divided into six categories according to the timing of appearance or disappearance during the 5-week period of comparison. Nineteen different proteins or protein fragments in 36 selected spots were identified by matrix-assisted laser-desorption ionization time of flight mass spectrometry (MS) or nano-electrospray tandem MS/MS. Some proteins were present throughout development (for example, cytosolic malate dehydrogenase), whereas others were associated with the early grain filling (ascorbate peroxidase) or desiccation (Cor14b) stages. Most noticeably, the development process is characterized by an accumulation of low-M(r) alpha-amylase/trypsin inhibitors, serine protease inhibitors, and enzymes involved in protection against oxidative stress. We present examples of proteins not previously experimentally observed, differential extractability of thiol-bound proteins, and possible allele-specific spot variation. Our results both confirm and expand on knowledge gained from previous analyses of individual proteins involved in grain filling and maturation.

  8. Isozyme expression of Chinese and Japanese populations of Chlamys farreri and their reciprocal hybrids

    Institute of Scientific and Technical Information of China (English)

    LI Taiwu; LIU Yan; SONG Linsheng; SUN Xiuqin

    2005-01-01

    Chinese and Japanese population of Chlamysfarreri and their reciprocal hybrids were surveyed in isozyme variability at 13 loci by polyacrytamide gel electrophoresis (PAGE). Isozyme banding patterns indicated these hybrids were diploid.Loci that were observed as being monomorphic in inbred populations of C. farreri were also found to be monomorphic in filial progeny;loci that observed to be polymorphic in parental type generations were also polymorphic in hybrid generations.Differences existed among allelic frequency of the four types of cross.Within the reciprocal hybrids the expression of malic enzyme (ME) isozyme was sufficient to distinguishing individual hybrids because of the band, Rf=0.38. However, there were no noticeable variations among all the samples to differentiate one from another. Inbreeding was likely to be the main problem in aquaculture. The introduction of new broodstock can improve the genetic diversity. Hybrid vigor has manifested to a certain extent in the present study.

  9. Conversion of crude Jatropha curcas seed oil into biodiesel using liquid recombinant Candida rugosa lipase isozymes.

    Science.gov (United States)

    Kuo, Ting-Chun; Shaw, Jei-Fu; Lee, Guan-Chiun

    2015-09-01

    The versatile Candida rugosa lipase (CRL) has been widely used in biotechnological applications. However, there have not been feasibility reports on the transesterification of non-edible oils to produce biodiesel using the commercial CRL preparations, mixtures of isozymes. In the present study, four liquid recombinant CRL isozymes (CRL1-CRL4) were investigated to convert various non-edible oils into biodiesel. The results showed that recombinant CRL2 and CRL4 exhibited superior catalytic efficiencies for producing fatty acid methyl ester (FAME) from Jatropha curcas seed oil. A maximum 95.3% FAME yield was achieved using CRL2 under the optimal conditions (50 wt% water, an initial 1 equivalent of methanol feeding, and an additional 0.5 equivalents of methanol feeding at 24h for a total reaction time of 48 h at 37 °C). We concluded that specific recombinant CRL isozymes could be excellent biocatalysts for the biodiesel production from low-cost crude Jatropha oil.

  10. Lignin peroxidase functionalities and prospective applications.

    Science.gov (United States)

    Falade, Ayodeji O; Nwodo, Uchechukwu U; Iweriebor, Benson C; Green, Ezekiel; Mabinya, Leonard V; Okoh, Anthony I

    2017-02-01

    Ligninolytic extracellular enzymes, including lignin peroxidase, are topical owing to their high redox potential and prospective industrial applications. The prospective applications of lignin peroxidase span through sectors such as biorefinery, textile, energy, bioremediation, cosmetology, and dermatology industries. The litany of potentials attributed to lignin peroxidase is occasioned by its versatility in the degradation of xenobiotics and compounds with both phenolic and non-phenolic constituents. Over the years, ligninolytic enzymes have been studied however; research on lignin peroxidase seems to have been lagging when compared to other ligninolytic enzymes which are extracellular in nature including laccase and manganese peroxidase. This assertion becomes more pronounced when the application of lignin peroxidase is put into perspective. Consequently, a succinct documentation of the contemporary functionalities of lignin peroxidase and, some prospective applications of futuristic relevance has been advanced in this review. Some articulated applications include delignification of feedstock for ethanol production, textile effluent treatment and dye decolourization, coal depolymerization, treatment of hyperpigmentation, and skin-lightening through melanin oxidation. Prospective application of lignin peroxidase in skin-lightening functions through novel mechanisms, hence, it holds high value for the cosmetics sector where it may serve as suitable alternative to hydroquinone; a potent skin-lightening agent whose safety has generated lots of controversy and concern. © 2016 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  11. Independent evolution of four heme peroxidase superfamilies.

    Science.gov (United States)

    Zámocký, Marcel; Hofbauer, Stefan; Schaffner, Irene; Gasselhuber, Bernhard; Nicolussi, Andrea; Soudi, Monika; Pirker, Katharina F; Furtmüller, Paul G; Obinger, Christian

    2015-05-15

    Four heme peroxidase superfamilies (peroxidase-catalase, peroxidase-cyclooxygenase, peroxidase-chlorite dismutase and peroxidase-peroxygenase superfamily) arose independently during evolution, which differ in overall fold, active site architecture and enzymatic activities. The redox cofactor is heme b or posttranslationally modified heme that is ligated by either histidine or cysteine. Heme peroxidases are found in all kingdoms of life and typically catalyze the one- and two-electron oxidation of a myriad of organic and inorganic substrates. In addition to this peroxidatic activity distinct (sub)families show pronounced catalase, cyclooxygenase, chlorite dismutase or peroxygenase activities. Here we describe the phylogeny of these four superfamilies and present the most important sequence signatures and active site architectures. The classification of families is described as well as important turning points in evolution. We show that at least three heme peroxidase superfamilies have ancient prokaryotic roots with several alternative ways of divergent evolution. In later evolutionary steps, they almost always produced highly evolved and specialized clades of peroxidases in eukaryotic kingdoms with a significant portion of such genes involved in coding various fusion proteins with novel physiological functions.

  12. Actinobacterial peroxidases: an unexplored resource for biocatalysis.

    Science.gov (United States)

    le Roes-Hill, Marilize; Khan, Nuraan; Burton, Stephanie Gail

    2011-07-01

    Peroxidases are redox enzymes that can be found in all forms of life where they play diverse roles. It is therefore not surprising that they can also be applied in a wide range of industrial applications. Peroxidases have been extensively studied with particular emphasis on those isolated from fungi and plants. In general, peroxidases can be grouped into haem-containing and non-haem-containing peroxidases, each containing protein families that share sequence similarity. The order Actinomycetales comprises a large group of bacteria that are often exploited for their diverse metabolic capabilities, and with recent increases in the number of sequenced genomes, it has become clear that this metabolically diverse group of organisms also represents a large resource for redox enzymes. It is therefore surprising that, to date, no review article has been written on the wide range of peroxidases found within the actinobacteria. In this review article, we focus on the different types of peroxidases found in actinobacteria, their natural role in these organisms and how they compare with the more well-described peroxidases. Finally, we also focus on work remaining to be done in this research field in order for peroxidases from actinobacteria to be applied in industrial processes.

  13. A new versatile peroxidase from Pleurotus.

    Science.gov (United States)

    Ruiz-Dueñas, F J; Camarero, S; Pérez-Boada, M; Martínez, M J; Martínez, A T

    2001-05-01

    Lignin peroxidase (LiP) and manganese peroxidase (MnP) have been investigated in Phanerochaete chrysosporium. A third ligninolytic peroxidase has been described in Pleurotus and Bjerkandera. Two of these versatile peroxidases (VPs) have been cloned, sequenced and characterized. They have high affinity for Mn(2+), hydroquinones and dyes, and also oxidize veratryl alcohol, dimethoxybenzene and lignin dimers. The deduced sequences show higher identity with Ph. chrysosporium LiP than MnP, but the molecular models obtained include a Mn(2+)-binding site. Concerning aromatic substrate oxidation, Pl. eryngii VP shows a putative long-range electron transfer pathway from an exposed trytophan to haem. Mutagenesis and chemical modification of this tryptophan and the acidic residues forming the Mn(2+)-binding site confirmed their role in catalysis. The existence of several substrate oxidation sites is supported further by biochemical evidence. Residue conservation in other fungal peroxidases is discussed.

  14. 紫外线照射对棉铃虫成虫几种同工酶的影响%Effects of ultraviolet light irradiation on several isozymes in Helicoverpa armigera (Hiiber) adults

    Institute of Scientific and Technical Information of China (English)

    孟建玉; 张长禹; 雷朝亮

    2012-01-01

    The effects of ultraviolet (UV) light stress on esterase, peroxidases (POX) , and catalase (CAT) isozymes in Helicoverpa armigera (Hüber) adults were studied by isozyme electrophoresis. When exposed to UV light irradiation, zymogram of esterase isozyme changed mainly in number and activity of isozyme. After 30 min and 60 min exposure, the intensity of isozyme bands E4,E9 and E10 were enhanced, E2 and E8 were weakened. The bands E1, E5, E7 and El 1 disappeared after UV light irradiation, while E3 and E6 newly emerged. At the longest exposure time (90 min), the intensity of isozyme bands E4 and E9 was enhanced, while the intensity of E2 and E8 was weakened. The bands El, E5 and E7 disappeared after UV light irradiation, whereas E3 and E6 newly emerged. The intensity of POX band P5 was enhanced in adults following the exposure to UV light for 30,60,90 minutes. The intensity of CAT band Cl was enhanced in adults following the exposure to UV light for 30,60,90 minutes, but that of band C2 was weakened after 30 min and 90 min exposure in comparison with the control.%采用同工酶电泳的方法,研究了紫外(ultraviolet,UV)胁迫条件下对棉铃虫体内酯酶、过氧化物酶(peroxidase,POX)及过氧化氢酶(catalase,CAT)等同工酶的影响.结果表明:与对照相比,紫外线照射处理组酯酶同工酶谱带发生了质和量的变化,照射时间为30 min和60 min时,谱带E4、E9、E10增强,谱带E2、E8减弱,谱带E1、F5、E7、E11消失,新增了谱带E3、E6;照射时间延长至90 min时,谱带F4、E9增强,谱带E2、E8减弱,谱带E1、E5、E7消失,新增了谱带E3、E6.紫外线照射处理下,棉铃虫POX同工酶谱带P5有所增强,CAT同工酶谱带的变化因照射时间不同而有所差异.与对照相比,紫外线照射处理组的C1谱带增强,但C2谱带在30 min时减弱,60 min时恢复到对照水平,90 min时与对照相比又有所减弱.

  15. Type correlation analysis between six morphological trait index and eight isozyme loci

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    @@ Indica-japonica differentiation is the majority for differentiation of Asian cultivated rice(O. Sativa L.). Sun et al proposed to distinguish indica and japonica by using the parameter Pj value which was quantified from six isozyme loci associated with indica and japonica differentiation, and the classification was consistent with the method of Cheng's "six combined morphological trait index" (CMT index). In this study, we analyzed the correlation between the six morphological traits and eight isozyme markers for indica-japonica classification by using 100 rice lines.

  16. Differential expression of 5-alpha reductase isozymes in the prostate and its clinical implications

    Directory of Open Access Journals (Sweden)

    Kai Wang

    2014-04-01

    Full Text Available The development of human benign or malignant prostatic diseases is closely associated with androgens, primarily testosterone (T and dihydrotestosterone (DHT. T is converted to DHT by 5-alpha reductase (5-AR isozymes. Differential expression of 5-AR isozymes is observed in both human benign and malignant prostatic tissues. 5-AR inhibitors (5-ARI are commonly used for the treatment of benign prostatic hyperplasia (BPH and were once promoted as chemopreventive agents for prostate cancer (PCa. This review discusses the role of the differential expression of 5-AR in the normal development of the human prostate and in the pathogenesis and progression of BPH and PCa.

  17. Aspects of gene structure and functional regulation of the isozymes of Na,K-ATPase

    DEFF Research Database (Denmark)

    Jorgensen, P.L.

    2001-01-01

    genomes, the genes of four alpha-subunit and at least three beta-subunit isoforms of Na,K-ATPase are identified and two gamma-subunits are expressed in kidney. The isoforms combine in a number of Na,K-ATPase isozymes that are expressed in a tissue and cell specific manner. Models of the molecular...... mechanism of regulation of these isozymes have become more reliable due to progress in understanding the three-dimensional protein structure and conformational transitions mediating transfer of energy from the P-domain to intramembrane Na+ and K+ binding sites....

  18. Triple Hybridization with Cultivated Barley (Hordeum vulgare L.)

    DEFF Research Database (Denmark)

    Bothmer, R. von; Claesson, L.; Flink, J.;

    1989-01-01

    represented species closely or distantly related to H. jubatum and H. lechleri. In trispecific crosses with diploid barley, the seed set was 5.7%. Crosses with tetraploid barley were highly unsuccessful (0.2% seed set). Three lines of diploid barley were used in the crosses, i.e. 'Gull', 'Golden Promise...

  19. Effects of n-butanol on barley microspore embryogenesis

    DEFF Research Database (Denmark)

    Castillo, Ana Maria; Nielsen, Nanna; Jensen, Anni

    2014-01-01

    Doubled haploid (DH) production is an efficient tool in barley breeding, but efficiency of DH methods is not consistent. Hence, the aim of this study was to study the effect of n-butanol application on DH barley plant production efficiency. Five elite cultivars of barley and thirteen breeding cro...

  20. MS Based Imaging of Barley Seed Development

    Institute of Scientific and Technical Information of China (English)

    Manuela Peukert; Andrea Matros; Hans-Peter Mock

    2012-01-01

    Spatially resolved analysis of metabolites and proteins is essential to model compartmentalized cellular processes in plants.Within recent years,tremendous progress has been made in MS based imaging (MSI) techniques,mostly MALDI MSI.The technology has been pioneered and is now widely applied in medicinal and pharmacological studies,and in recent years found its way into plant science (Kaspar et al.,2011; Peukert etal.,2012).We are interested in the elucidation of spatially resolved metabolic networks related to barley grain development.An understanding of developmentally and ecologically regulated processes affecting agronomical traits such as final grain weight,seed quality and stress tolerance is of outmost importance,as barley provides one of the staple foods.Barley also serves as a model plant for other cereals such as wheat.The presentation will introduce an untargeted MALDI MSI approach to the analysis of me-tabolite patterns during barley grain development.We analyzed longitudinal and cross sections from developing barley grains (3,7,10 and 14 days after pollination).In the presentation we will address spatial resolution,sensitivity and identification of unknown compounds will also be discussed.A major task is to connect the metabolite patterns to distinct cellular and physiological events.As an example,particular metabolite distributions indicative for nutrient transport into the developing endosperm will be shown.

  1. Evaluation of Barley as Human Food

    Directory of Open Access Journals (Sweden)

    Mehmet Köten

    2013-12-01

    Full Text Available Barley, as animal feed, raw material for malting and human food, constitute an important part among cereal sources in the world. Majority of barley that produced both in Turkey and other countries of the world, is being used as animal feed. Poor baking quality, taste and appearance of barley restricted its use in human nutrition. However, recently high protein, fiber, especially β-glucan and high starch content appeal to food industry. Many scientific researches established that β-glucan, a soluble fiber, has an effect in healing coronary-hearth diseases, lowering blood cholesterol level, balancing blood sugar level, preventing obesity. Being a healthy cereal that can be used in various purposes, and an additive in many food products, barley is considered a very promising cereal, and research to increase possibilities of its use in human nutrition is being increased. In the literature, there has been researches on making noodles, bulgur, kavut (roasted cereal, breakfast cereals. In this study the researches relating to evaluation of barley, importance of which is increased every day, as human food was reviewed.

  2. No effect of 5-fluorouracil on the properties of purified. cap alpha. -amylase from barley half-seeds

    Energy Technology Data Exchange (ETDEWEB)

    Rodaway, S.J.; Kende, H.

    1978-01-01

    Amylase has been purified from de-embryonated seeds of barley (Horedeum vulgare L. cv. Betzes) which have been incubated on 10/sup 6/M gibberellic acid (GA/sub 3/) following 3 days of imbibition in buffer. Incubation of the half-seeds in up to 10/sup -2/ M 5-fluorouracil (5-FU) during the entire incubation period, including imbibition, had no effect on any of the following characteristics of purified ..cap alpha..-amylase: thermal stability in the absence of calcium, molecular weight of the enzyme, isozyme composition, specific activity, or the amount of ..cap alpha..-amylase synthesized by the aleurone tissue. The synthesis of rRNA and tRNA was strongly inhibited by 5-FU, indicating that the analog had entered the aleurone cells. These results are not in agreement with those of Carlson (Nature New Biology 237: 39-41 (1972)) who found that treatment of barley aleurone with 10/sup -4/ M 5-FU to the addition of GA/sub 3/ resulted in decreased thermal stability of GA/sub 3/-induced ..cap alpha..-amylase and who interpreted this as evidence that the mRNA for ..cap alpha..-amylase was synthesized during the imbibition of the aleurone tissue and independently of gibberellin action. Results of the present experiments indicate that the thermal stability of highly purified ..cap alpha..-amylase is not altered by treatment of barley half-seeds with 5-FU, and that 5-FU cannot be used as a probe to examine the timing of ..cap alpha..-amylase mRNA synthesis.

  3. Analysis on Genetic Diversity of Bougainvillea spp.Germplasm Resources Based on Isozyme%宝巾(Bougainvillea spp.)种质资源同工酶的遗传多样性分析

    Institute of Scientific and Technical Information of China (English)

    陈庭; 刘伟; 谢良生; 雷江丽

    2012-01-01

    [Objective] The aim was to analyze the genetic adversity of 12 Bougainvillea spp. Germplasm resources from Shenzhen region. [Method] The esterase (EST) isozyme and peroxidase (POD) isozyme of Bougainvillea spp. Germplasm resources were determined by poly-acrylamide gel electrophoresis and the cluster analysis was used according to statistical results. [ Result ] There was great genetic adversity a-mong different germplasm resources and the resources could be divided into two groups based on 0.63 of similarity coefficient. [Conclusion] The characteristic band of isozyme was suitable for variety identification and defining the blood relationship for various varieties to some extent.%[目的]分析深圳本地12种宝巾种质资源的遗传多样性.[方法]利用聚丙烯酰胺凝胶电泳法研究宝巾种质资源过氧化物同工酶(POD)和酯酶同工酶(EST)的遗传多样性,并对统计结果进行聚类分析.[结果]各宝巾种质资源间有丰富的遗传多样性,在相似系数为0.63水平上可以聚为2大类.[结论]同工酶的特征谱带应用于品种鉴定是可行的,在一定程度上明确了各宝巾品种的亲缘关系.

  4. Immigration of the barley mildew pathogen into field plots of barley

    DEFF Research Database (Denmark)

    O'Hara, R.B.; Brown, J.K.M.

    1996-01-01

    Immigration of the barley powdery mildew pathogen (Erysiphe graminis f.sp. hordei) into field plots of the spring barley variety Tyra (carrying the resistance allele Mla1) was investigated. Spores were trapped from the top of the plot canopies, as well as from control plots of wheat with no barley...... nearby. Comparison of the frequencies of virulent and avirulent single-colony isolates showed that the amount of immigration, relative to the amount of inoculum being produced within the plot, reduced very rapidly, until it could not be detected in the middle of the growing season (mid-June)....

  5. Glycosylation and thermodynamic versus kinetic stability of horseradish peroxidase

    DEFF Research Database (Denmark)

    Tams, J.W.; Welinder, Karen G.

    1998-01-01

    Glycoprotein stability, glycoprotein unfolding, horseradish peroxidase, thermodynamic stability, kinetik stability......Glycoprotein stability, glycoprotein unfolding, horseradish peroxidase, thermodynamic stability, kinetik stability...

  6. Selenium, glutathione peroxidase and other selenoproteins

    Energy Technology Data Exchange (ETDEWEB)

    Wilhelmsen, E.C.

    1983-01-01

    Selenium, as essential trace element, has long been associated with protein. The essentiality of selenium is partially understood as glutathione peroxidase contains an essential selenocysteine. Glutathione peroxidase has been purified from many tissues including rat liver. An estimated molecular weight of 105,000 was obtained for glutathione peroxidase by comparison to standards. A subunit size of 26,000 was obtained by SDS-gel electrophoresis. Glutathione peroxidase is not the only selenoprotein in the rat. In seven rat tissues examined, there were many different subunit sizes and change groups representing between 9 and 23 selenoproteins. Selenocysteine in glutathione peroxidase accounts for ca. 36% of the selenium in the rat. The mode of synthesis of glutathione peroxidase and the other selenoproteins is not understood. Glutathione peroxidase is strongly and reversibly inhibited by mercaptocarboxylic acids and other mercaptans, including some used as slow-acting drugs for the symtomatic treatment of rheumatoid arthritis. The mechanism and chemistry of this inhibition is discussed. This inhibition may provide a link between selenium and arthritis.

  7. Thiol-Based Peroxidases and Ascorbate Peroxidases: Why Plants Rely on Multiple Peroxidase Systems in the Photosynthesizing Chloroplast?

    Science.gov (United States)

    Dietz, Karl-Josef

    2016-01-01

    Photosynthesis is a highly robust process allowing for rapid adjustment to changing environmental conditions. The efficient acclimation depends on balanced redox metabolism and control of reactive oxygen species release which triggers signaling cascades and potentially detrimental oxidation reactions. Thiol peroxidases of the peroxiredoxin and glutathione peroxidase type, and ascorbate peroxidases are the main peroxide detoxifying enzymes of the chloroplast. They use different electron donors and are linked to distinct redox networks. In addition, the peroxiredoxins serve functions in redox regulation and retrograde signaling. The complexity of plastid peroxidases is discussed in context of suborganellar localization, substrate preference, metabolic coupling, protein abundance, activity regulation, interactions, signaling functions, and the conditional requirement for high antioxidant capacity. Thus the review provides an opinion on the advantage of linking detoxification of peroxides to different enzymatic systems and implementing mechanisms for their inactivation to enforce signal propagation within and from the chloroplast.

  8. Nitration of Phenol Catalyzed by Horseradish Peroxidase

    Institute of Scientific and Technical Information of China (English)

    DAI Rong-ji; HUANG Hui; TONG Bin; XIAO Sheng-yuan

    2007-01-01

    Horseradish peroxidase, an acidic peroxidase from the horseradish, is one of the most important enzymes as analytical reagent.The enzymatic nitration of phenol by oxidation of nitrite was studied using horseradish peroxidase in the presence of H2O2.The results showed that nitration occur at 2- and 4- positions of phenol.There were also minor products of hydroquinone and catechol.The influence of various reaction parameters, including pH, organic solvent, and concentration of H2O2, on nitration products were discussed.The best nitration pH was 7.0, and H2O2 should be added to the reaction mixture slowly.

  9. Morphological and Isozyme Variation in Natural Populations of the Genus Medicago L. Prospected in Northern Algeria

    Directory of Open Access Journals (Sweden)

    Imane MEDOUKALI

    2015-04-01

    Full Text Available As part of the evaluation and enhancement of genetic resources, morphological and isozyme variability within and among 169 accessions, representing 14 species of the genus Medicago L. collected in northern Algeria, was assessed using twelve quantitative traits and two enzymatic systems. Phenotype frequencies were scored in six enzyme zones to determine isozyme variability within and among populations. The data analysis resolved a high level of genetic diversity. Ten morphometric characteristics contributed to the discrimination of the species. The relationship between the collection site environment and phenotypic characteristics was also studied. Esterase (EST enzyme system was more polymorphic than glutamate oxaloacetate transaminase (GOT system. Were scored 2 zones with 10 bands and 21 phenotypes for GOT (glutamate oxaloacetate transaminase and 4 zones with 22 bands and 71 phenotypes for EST (esterase Polymorphism index and Jaccard’s genetic distances revealed the existence of a high genetic diversity within and among the studied populations. The annual species M. polymorpha presented an intraspecific polymorphism index of 0.57, which was higher than all other species indices. Clustering of the species based on isozyme markers was in agreement with taxonomic criteria and showed no significant correlation with morphological characteristics. Conservation programs should take into account the level of genetic diversity within and between populations revealed by isozyme markers.

  10. CHARACTERIZATION OF LACTATE DEHYDROGENASE ISOZYME PATTERN AND MORPHOLOGY OF THREE MARINE FISH CELL LINES

    Institute of Scientific and Technical Information of China (English)

    郭华荣; 张士璀; 李红岩; 童裳亮; 相建海

    2002-01-01

    Three continuous marine fish cell lines of FG (i.e. , Flounder Gill) from flounder (Paralichthys olivaceus) gill, SPH (i.e., Sea Perch Heart) fro m sea perch (Lateolabrax japonicus) heart and RSBF (i.e., Red Sea Bream Fin) from red se a bream (Pagrosomus major) fin, were characterized by lactate dehydrogenase (LDH) is ozyme and morphological analysis. The LDH isozyme patterns of these three cell lines and their corresponding tissues of origin were investigated and compared. The results sho wed: (1) No difference was found in the LDH isozyme patterns of FG and flounder gill tissue. However, the LDH isozyme patterns of SPH and RSBF were significantly different from their cor responding tissues of origin; (2) LDH isozyme patterns of FG, SPH and RSBF were markedly di fferent from each other and could serve as genetic markers for species identification and de tection of cross contamination. Morphological change analysis of these three cell lines in compa rison to their original tissues indicated that FG cells still appeared epithelioid without mor phological transformation. However, morphological changes were found in SPH and RSBF compa red to their original tissues. Therefore, the cellular morphology was still plastic in the relatively stable culture conditions, and it was possible that change of LDH patterns was related to morphological changes of fish cells in vitro.

  11. Analyzing genetic diversity in conifers...isozyme resolution by starch gel electrophoresis

    Science.gov (United States)

    M. Thompson Conkle

    1972-01-01

    Enzymes in forest tree materials can be resolved by starch gel electrophoresis. A gel slab is prepared in a mold assembled from glass and plastic. Wicks containing an aqueous extract of macerated plant material are inserted in the gel and processed. The gel is sliced, stained, examined, and photographed. Isozyme bands produced by differential migration of enzymes...

  12. Enzymatic properties of chorismate synthase isozymes of tomato (Lycopersicon esculentum Mill.).

    Science.gov (United States)

    Braun, M; Henstrand, J M; Görlach, J; Amrhein, N; Schmid, J

    1996-01-01

    Three plastidic chorismate synthase isozymes (CS1, CS2 and CS2 delta) of tomato were identified by isolation of the corresponding cDNAs. These three cDNAs are derived from only two genes (LeCS1 and LeCS2). This additional complexity results from differential splicing of the primary transcript of one of the genes (LeCS2) giving rise to two different transcripts (CS2 and CS2 delta transcripts). All three isozymes were individually expressed in Escherichia coli both as precursor proteins with N-terminal transit peptides and as mature proteins. Only the mature but not the precursor isozymes CS1 and CS2 were enzymatically active. The enzyme CS2 delta was unstable in E. coli. Both CS1 and CS2 were purified to near homogeneity and their enzymatic properties were analyzed. They differ substantially in their Km values for the substrate 5-enol-pyruvylshikimate 3-phosphate (11 and 80 microM for the mature forms of CS1 and CS2, respectively). The two isozymes appear to be active only as oligomers, and the potential physiological implications of this result are discussed.

  13. Mechanism of phosphorylation-induced activation of phospholipase C-gamma isozymes.

    Science.gov (United States)

    Gresset, Aurelie; Hicks, Stephanie N; Harden, T Kendall; Sondek, John

    2010-11-12

    The lipase activity of most phospholipases C (PLCs) is basally repressed by a highly degenerate and mostly disordered X/Y linker inserted within the catalytic domain. Release of this auto-inhibition is driven by electrostatic repulsion between the plasma membrane and the electronegative X/Y linker. In contrast, PLC-γ isozymes (PLC-γ1 and -γ2) are structurally distinct from other PLCs because multiple domains are present in their X/Y linker. Moreover, although many tyrosine kinases directly phosphorylate PLC-γ isozymes to enhance their lipase activity, the underlying molecular mechanism of this activation remains unclear. Here we define the mechanism for the unique regulation of PLC-γ isozymes by their X/Y linker. Specifically, we identify the C-terminal SH2 domain within the X/Y linker as the critical determinant for auto-inhibition. Tyrosine phosphorylation of the X/Y linker mediates high affinity intramolecular interaction with the C-terminal SH2 domain that is coupled to a large conformational rearrangement and release of auto-inhibition. Consequently, PLC-γ isozymes link phosphorylation to phospholipase activation by elaborating upon primordial regulatory mechanisms found in other PLCs.

  14. Studies of the role of selenium-independent and selenium-dependent glutathione peroxidases in eicosanoid biosynthesis

    Energy Technology Data Exchange (ETDEWEB)

    Zisman, A.H.

    1990-01-01

    Glutathione S-transferases are involved in the biotransformation and/or detoxification of a wide range of organic compounds, including allylic epoxides. GSTs catalyze the transformation of prostaglandin (PG)H[sub 2] into PGE[sub 2] and/or PGF[sub 2alpha]. Specific GST isozymes possessing non-selenium glutathione-peroxidase activity (NonSe-GSH-PX) catalyze the direct reduction of PGH[sub 2] to PGF[sub 2alpha]. Other GST isozymes have been reported to catalyze the transformation of leukotriene (LT)A[sub 4] into LTC[sub 4]. In this study, human liver GSTs were purified and individual isozymes were characterized by SDS electrophoresis, isoelectric focusing, substrate specificities, immunological cross reactivities, and their ability to catalyze the transformation of PGH[sub 2] to PGF[sub 2alpha], and LTA[sub 4] to LTC[sub 4]. The GST isozyme expression pattern in man varies between individuals. Se-dependent GSH-PX activity (Se-GSH-PX) also plays a role in eicosanoid metabolism. The author infused Se-adequate and Se-deficient dairy cattle with endotoxin into the mammary gland to simulate an inflammation. Arachidonic acid metabolites were extracted and analyzed in both the milk and the milk polymorphonuclear leukocytes (PMNs). PMN's cytosol was assayed for Se- and nonSe-GSH-PX, and GST activity. The results indicate that the Se-deficient cows had lower levels (p < 0.05) of PGE[sub 2] and TXB[sub 2] released into the milk following challenge; however, there was no significant effect on the arachidonic acid metabolites produced by the milk PMNs. Although the Se-deficient cows had significantly lower levels (p < 0.05) of Se-GSH-PX activity, there was no effect on the GST nor NonSe-GSH-PX activity. Overall, eicosanoid biosynthesis is complex, being influenced by both dietary and enzymatic manipulation. The data support Se- and nonSe-GSH-PX playing important roles in eicosanoid formation.

  15. Disulfide bonds and glycosylation in fungal peroxidases.

    Science.gov (United States)

    Limongi, P; Kjalke, M; Vind, J; Tams, J W; Johansson, T; Welinder, K G

    1995-01-15

    Four conserved disulfide bonds and N-linked and O-linked glycans of extracellular fungal peroxidases have been identified from studies of a lignin and a manganese peroxidase from Trametes versicolor, and from Coprinus cinereus peroxidase (CIP) and recombinant C. cinereus peroxidase (rCIP) expressed in Aspergillus oryzae. The eight cysteine residues are linked 1-3, 2-7, 4-5 and 6-8, and are located differently from the four conserved disulfide bridges present in the homologous plant peroxidases. CIP and rCIP were identical in their glycosylation pattern, although the extent of glycan chain heterogeneity depended on the fermentation batch. CIP and rCIP have one N-linked glycan composed only of GlcNAc and Man at residue Asn142, and two O-linked glycans near the C-terminus. The major glycoform consists of single Man residues at Thr331 and at Ser338. T. versicolor lignin isoperoxidase TvLP10 contains a single N-linked glycan composed of (GlcNAc)2Man5 bound to Asn103, whereas (GlcNAc)2Man3 was found in T. versicolor manganese isoperoxidase TvMP2 at the same position. In addition, mass spectrometry of the C-terminal peptide of TvMP2 indicated the presence of five Man residues in O-linked glycans. No phosphate was found in these fungal peroxidases.

  16. The barley Jip23b gene

    DEFF Research Database (Denmark)

    Müller-Uri, Frieder; Cameron-Mills, Verena; Mundy, John

    2002-01-01

    The barley gene (Jip23) encoding a 23,000-Da protein of unknown function was isolated and shown to be induced by jasmonate methyl ester (MeJA) in leaves. 5'upstream Jip23 sequence was isolated and fused to the beta-glucuronidase gene (GUS), and this reporter was introduced by particle bombardment...

  17. Cisgenic barley with improved phytase activity

    DEFF Research Database (Denmark)

    Holme, Inger; Dionisio, Giuseppe; Brinch-Pedersen, Henrik

    2010-01-01

    are accordingly very similar to those generated by conventional breeding. The cisgenesis concept allows for the introduction of extra gene copies of a particular gene to accentuate the trait. We are using a barley purple acid phosphatase expressed during grain filling as candidate gene for cisgenesis. A genomic...

  18. Endoproteolytic activity assay in malting barley

    Directory of Open Access Journals (Sweden)

    Blanca Gómez Guerrero

    2013-12-01

    Full Text Available Hydrolysis of barley proteins into peptides and amino acids is one of the most important processes during barley germination.The degradation of the endosperm stored proteins facilitates water and enzyme movements, enhances modification, liberates starch granules and increases soluble amino nitrogen. Protease activity is the result of the activities of a mixture of exo- and endo-proteases. The barley proteins are initially solubilized by endo-proteases and the further by exo-proteases. Four classes of endo-proteases have been described: serine-proteases, cysteine-proteases, aspartic-proteases and metallo-proteases. The objective of this work was to develop a rapid and colorimetric enzymatic assay to determine the endo-proteolytic activity of the four endo-protease classes using two different substrates: azo-gelatin and azo-casein. Optimum conditions for the assays such as: pH,reaction time and temperature and absorbance scale were determined. Azo-gelatin presented several difficulties in standardizing an “in solution” assay. On the other hand, azo-casein allowed standardization of the assay for the four enzyme classes to produce consistent results. The endo-proteoteolytic method developed was applied to determine the endo-protease activity in barley, malt and wort.

  19. Adaptation of barley to harsh Mediterranean environments.

    NARCIS (Netherlands)

    Oosterom, van E.

    1993-01-01

    Research ObjectivesBarley is in Syria the dominant crop in areas receiving less than 300 mm annual precipitation. Grain yield is often below 1 ton ha -1, and is reduced by low temperatures in winter and terminal drought stress in spring. Variation i

  20. 21 CFR 864.7675 - Leukocyte peroxidase test.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Leukocyte peroxidase test. 864.7675 Section 864... peroxidase test. (a) Identification. A leukocyte peroxidase test is a device used to distinguish certain... peroxidase activity as evidenced by staining. The results of this test are used in the differential...

  1. Effects of Waterlogging on Photosynthesis and Antioxidant Enzyme Activities of Six Barley Genotypes with Different Waterlogging Tolerance

    Institute of Scientific and Technical Information of China (English)

    XIAO Yu-ping; WEI Kang; CHEN Jin-xin; ZHOU Mei-xue; ZHANG Guo-ping

    2005-01-01

    A field experiment was carried out to study genotypic difference in the effect of waterlogging on photosynthesis, chlorophyll content and antioxidative enzyme activities in barley. Waterlogging caused a rapid decline in net photosynthetic rate (Ph) and stomatal conductance (gs), and little change in chlorophyll content during early days of the treatment. A dramatic increase in malondialdehyde (MDA) content, superoxide dismutase (SOD) and peroxidase (POD) in waterlogged plants in the early days of the experiment was found, indicating the occurrence of oxidative stress in barley plants exposed to waterlogging. There was a highly significant difference in the changed extent of all these parameters among genotypes.Franklin and Yongjiahong Liuleng Damai, which were relatively sensitive to waterlogging in terms of growth, photosynthesis and chlorophyll content, accumulated much more MDA than the other two relatively tolerant genotypes (93-3143 and QS).After removal of waterlogging, the genotypic difference became much greater in recovering of these examined parameters.Yongjiahong Liuleng Damai showed higher recovery, while Franklin only recovered to 50% of the control at the 14 day after waterlogging removal. It may be concluded that it is the difference in anti-oxidative stress caused by waterlogging that account for the major difference in photosynthesis among barley genotypes.

  2. Quantitative analysis of proteome extracted from barley crowns grown under different drought conditions.

    Science.gov (United States)

    Vítámvás, Pavel; Urban, Milan O; Škodáček, Zbynek; Kosová, Klára; Pitelková, Iva; Vítámvás, Jan; Renaut, Jenny; Prášil, Ilja T

    2015-01-01

    Barley cultivar Amulet was used to study the quantitative proteome changes through different drought conditions utilizing two-dimensional difference gel electrophoresis (2D-DIGE). Plants were cultivated for 10 days under different drought conditions. To obtain control and differentially drought-treated plants, the soil water content was kept at 65, 35, and 30% of soil water capacity (SWC), respectively. Osmotic potential, water saturation deficit, (13)C discrimination, and dehydrin accumulation were monitored during sampling of the crowns for proteome analysis. Analysis of the 2D-DIGE gels revealed 105 differentially abundant spots; most were differentially abundant between the controls and drought-treated plants, and 25 spots displayed changes between both drought conditions. Seventy-six protein spots were successfully identified by tandem mass spectrometry. The most frequent functional categories of the identified proteins can be put into the groups of: stress-associated proteins, amino acid metabolism, carbohydrate metabolism, as well as DNA and RNA regulation and processing. Their possible role in the response of barley to drought stress is discussed. Our study has shown that under drought conditions barley cv. Amulet decreased its growth and developmental rates, displayed a shift from aerobic to anaerobic metabolism, and exhibited increased levels of several protective proteins. Comparison of the two drought treatments revealed plant acclimation to milder drought (35% SWC); but plant damage under more severe drought treatment (30% SWC). The results obtained revealed that cv. Amulet is sensitive to drought stress. Additionally, four spots revealing a continuous and significant increase with decreasing SWC (UDP-glucose 6-dehydrogenase, glutathione peroxidase, and two non-identified) could be good candidates for testing of their protein phenotyping capacity together with proteins that were significantly distinguished in both drought treatments.

  3. QUANTITATIVE ANALYSIS OF PROTEOME EXTRACTED FROM BARLEY CROWNS GROWN UNDER DIFFERENT DROUGHT CONDITIONS

    Directory of Open Access Journals (Sweden)

    Pavel eVítámvás

    2015-06-01

    Full Text Available Barley cv. Amulet was used to study the quantitative proteome changes through different drought conditions utilizing two-dimensional difference gel electrophoresis (2D-DIGE. Plants were cultivated for ten days under different drought conditions. To obtain control and differentially drought-treated plants, the soil water content was kept at 65%, 35%, and 30% of soil water capacity (SWC, respectively. Osmotic potential, water saturation deficit, 13C discrimination, and dehydrin accumulation were monitored during sampling of the crowns for proteome analysis. Analysis of the 2D-DIGE gels revealed 105 differentially abundant spots; most were differentially abundant between the controls and drought-treated plants, and 25 spots displayed changes between both drought conditions. Seventy-six protein spots were successfully identified by tandem mass spectrometry. The most frequent functional categories of the identified proteins can be put into the groups of: stress-associated proteins, amino acid metabolism, carbohydrate metabolism, as well as DNA & RNA regulation and processing. Their possible role in the response of barley to drought stress is discussed. Our study has shown that under drought conditions barley cultivar Amulet decreased its growth and developmental rates, displayed a shift from aerobic to anaerobic metabolism, and exhibited increased levels of several protective proteins. Comparison of the two drought treatments revealed plant acclimation to milder drought (35% SWC; but plant damage under more severe drought treatment (30% SWC. The results obtained revealed that cv. Amulet is sensitive to drought stress. Additionally, four spots revealing a continuous and significant increase with decreasing SWC (UDP-glucose 6-dehydrogenase, glutathione peroxidase, and two non-identified could be good candidates for testing of their protein phenotyping capacity together with proteins that were significantly distinguished in both drought treatments.

  4. Characterization of barley serpin Z7 that plays multiple roles in malt and beer.

    Science.gov (United States)

    Li, Xiaomin; Jin, Zhao; Gao, Fei; Lu, Jian; Cai, Guolin; Dong, Jianjun; Yu, Junhong; Yang, Mei

    2014-06-18

    Barley protein Z7 (BSZ7) is a well-known serine protease inhibitor that was regarded as a major effector of beer foam stability. Moreover, it has also been suggested to participate in haze formation and affect wort filterability. The present study purified BSZ7 from barley malt and characterized its secondary structure and modification, as well as its relationship with peroxidase, to elucidate the molecular base of BSZ7 that supports its multiple roles in malt and beer. It was found that after 30 min of heating, the secondary structure was not affected. BSZ7 has no inhibiting effect on nonspecific protease originated from malt, suggesting its negative role in wort filterability was accomplished by other means. Furthermore, the glycation of BSZ7 by the Maillard reaction may make some contribution to its survival during wort boiling. The interaction of BSZ7 with polysaccharides and polyphenols found by adding experiment may explain how it acts as a negative factor on wort filterability. Greater understanding of BSZ7 and other proteins of malts will lead to better improvements in brewing quality.

  5. Transgenic barley: a prospective tool for biotechnology and agriculture.

    Science.gov (United States)

    Mrízová, Katarína; Holasková, Edita; Öz, M Tufan; Jiskrová, Eva; Frébort, Ivo; Galuszka, Petr

    2014-01-01

    Barley (Hordeum vulgare L.) is one of the founder crops of agriculture, and today it is the fourth most important cereal grain worldwide. Barley is used as malt in brewing and distilling industry, as an additive for animal feed, and as a component of various food and bread for human consumption. Progress in stable genetic transformation of barley ensures a potential for improvement of its agronomic performance or use of barley in various biotechnological and industrial applications. Recently, barley grain has been successfully used in molecular farming as a promising bioreactor adapted for production of human therapeutic proteins or animal vaccines. In addition to development of reliable transformation technologies, an extensive amount of various barley genetic resources and tools such as sequence data, microarrays, genetic maps, and databases has been generated. Current status on barley transformation technologies including gene transfer techniques, targets, and progeny stabilization, recent trials for improvement of agricultural traits and performance of barley, especially in relation to increased biotic and abiotic stress tolerance, and potential use of barley grain as a protein production platform have been reviewed in this study. Overall, barley represents a promising tool for both agricultural and biotechnological transgenic approaches, and is considered an ancient but rediscovered crop as a model industrial platform for molecular farming.

  6. A robust and extracellular heme-containing peroxidase from Thermobifida fusca as prototype of a bacterial peroxidase superfamily

    NARCIS (Netherlands)

    van Bloois, Edwin; Torres Pazmino, Daniel; Winter, Remko T.; Fraaije, Marco W.

    2010-01-01

    DyP-type peroxidases comprise a novel superfamily of heme-containing peroxidases which is unrelated to the superfamilies of known peroxidases and of which only a few members have been characterized in some detail. Here, we report the identification and characterization of a DyP-type peroxidase (TfuD

  7. [Cold induced cDNA library construction of highland barley (Hordeum vulgare L. var. nudum Hk. f.) using suppression subtractive hybridization technology].

    Science.gov (United States)

    He, Tao; Jia, Jing Fen

    2008-12-01

    Cold-induced genes of highland barley (Hordeum vulgare L. var. nudum Hk. f.) were studied using suppression subtractive hybridization (SSH) technique. The cDNA from the materials treated with 4 degrees C was used as "tester", and that from the materials growing in green house (20+/-2 degrees C) as "driver". A subtractive library of highland barley including 640 cDNA clones was constructed in this study. Enzyme digestion of 32 clones chosen randomly from the library indicated that 87.5% of them contained inserts. The cDNA inserts of 16 clones were sequenced. Blast search analyses showed that these cDNAs were homologies to genes encoding the following proteins: metallothionein, protein kinase, ethylene signal transcription factor, bZIP transcription factor, zing finger transcription factor, ribulose-1,5-bisphosphate carboxylase, ribosomal protein, sodium: hydrogen antiporter, catalase, NADPH-cytochrome reductase, ascorbate peroxidase, DNA binding protein, and sugar transporter-like protein. These results indicated that the cDNA clones in the library were related to cold-induced genes, and suggested that the cold-tolerant mechanism of highland barley might be a complicated, interactive system involving multiple approaches and genes. Construction of subtractive cDNA library provided an advantage for further studies to isolate and clone cold-induced genes in highland barley.

  8. Anionic peroxidase production by Arnebia euchroma callus.

    Science.gov (United States)

    Farhadi, Sahar; Haghbeen, Kamahldin; Marefatjo, Mohammad-Javad; Hoor, Marjan Ghiyami; Zahiri, Hossein Shahbani; Rahimi, Karim

    2011-01-01

    Arnebia euchroma callus, obtained from the root cell culture of an Iranian native specimen, has gained a doubling time of 63 H after regular subculturing on Linsmaier-Skoog (LS) medium containing sugar (50 g/L), 2,4-dichlorophenoxyacetic acid (10(-6) M), and kinetin (10(-5) M) under darkness at 25°C. Despite the observed somaclonal variations, peroxidase production by the A. euchroma calli has been stable over 4 years under the aforementioned conditions. Isoelectric focusing experiments revealed that the partially purified A. euchroma peroxidases (AePoxs) are mainly anionic with pI values of about 5.5 and 6.6. AePox reaches its optimal activity at 55°C and pH 7.5. Results of the various kinetic studies suggest that AePox belongs to the type III plant peroxidases with no activity for the oxidation of 3-indoleacetic acid, but seems to play a role in the lignin biosynthesis and H(2) O(2) regulation during the proliferation of the A. euchroma cells on LS medium. Comparing the biochemical properties of AePox with horseradish peroxidase and in view of the ease of solid cell culture, the A. euchroma callus could be considered as a source of plant peroxidase for some biotechnological applications. Copyright © 2011 International Union of Biochemistry and Molecular Biology, Inc.

  9. Association mapping of partitioning loci in barley

    Directory of Open Access Journals (Sweden)

    Mackay Ian J

    2008-02-01

    Full Text Available Abstract Background Association mapping, initially developed in human disease genetics, is now being applied to plant species. The model species Arabidopsis provided some of the first examples of association mapping in plants, identifying previously cloned flowering time genes, despite high population sub-structure. More recently, association genetics has been applied to barley, where breeding activity has resulted in a high degree of population sub-structure. A major genotypic division within barley is that between winter- and spring-sown varieties, which differ in their requirement for vernalization to promote subsequent flowering. To date, all attempts to validate association genetics in barley by identifying major flowering time loci that control vernalization requirement (VRN-H1 and VRN-H2 have failed. Here, we validate the use of association genetics in barley by identifying VRN-H1 and VRN-H2, despite their prominent role in determining population sub-structure. Results By taking barley as a typical inbreeding crop, and seasonal growth habit as a major partitioning phenotype, we develop an association mapping approach which successfully identifies VRN-H1 and VRN-H2, the underlying loci largely responsible for this agronomic division. We find a combination of Structured Association followed by Genomic Control to correct for population structure and inflation of the test statistic, resolved significant associations only with VRN-H1 and the VRN-H2 candidate genes, as well as two genes closely linked to VRN-H1 (HvCSFs1 and HvPHYC. Conclusion We show that, after employing appropriate statistical methods to correct for population sub-structure, the genome-wide partitioning effect of allelic status at VRN-H1 and VRN-H2 does not result in the high levels of spurious association expected to occur in highly structured samples. Furthermore, we demonstrate that both VRN-H1 and the candidate VRN-H2 genes can be identified using association mapping

  10. Application of repeated aspartate tags to improving extracellular production of Escherichia coli L-asparaginase isozyme II.

    Science.gov (United States)

    Kim, Sun-Ki; Min, Won-Ki; Park, Yong-Cheol; Seo, Jin-Ho

    2015-11-01

    Asparaginase isozyme II from Escherichia coli is a popular enzyme that has been used as a therapeutic agent against acute lymphoblastic leukemia. Here, fusion tag systems consisting of the pelB signal sequence and various lengths of repeated aspartate tags were devised to highly express and to release active asparaginase isozyme II extracellularly in E. coli. Among several constructs, recombinant asparaginase isozyme II fused with the pelB signal sequence and five aspartate tag was secreted efficiently into culture medium at 34.6 U/mg cell of specific activity. By batch fermentation, recombinant E. coli produced 40.8 U/ml asparaginase isozyme II in the medium. In addition, deletion of the gspDE gene reduced extracellular production of asparaginase isozyme II, indicating that secretion of recombinant asparaginase isozyme II was partially ascribed to the recognition by the general secretion machinery. This tag system composed of the pelB signal peptide, and repeated aspartates can be applied to extracellular production of other recombinant proteins.

  11. Barley grain for ruminants: A global treasure or tragedy

    Directory of Open Access Journals (Sweden)

    Nikkhah Akbar

    2012-07-01

    Full Text Available Abstract Barley grain (Hordeum vulgare L. is characterized by a thick fibrous coat, a high level of ß-glucans and simply-arranged starch granules. World production of barley is about 30 % of that of corn. In comparison with corn, barley has more protein, methionine, lysine, cysteine and tryptophan. For ruminants, barley is the third most readily degradable cereal behind oats and wheat. Due to its more rapid starch fermentation rate compared with corn, barley also provides a more synchronous release of energy and nitrogen, thereby improving microbial nutrient assimilation. As a result, feeding barley can reduce the need for feeding protected protein sources. However, this benefit is only realized if rumen acidity is maintained within an optimal range (e.g., > 5.8 to 6.0; below this range, microbial maintenance requirements and wastage increase. With a low pH, microbial endotoxines cause pro-inflammatory responses that can weaken immunity and shorten animal longevity. Thus, mismanagement in barley processing and feeding may make a tragedy from this treasure or pearl of cereal grains. Steam-rolling of barley may improve feed efficiency and post-rumen starch digestion. However, it is doubtful if such processing can improve milk production and feed intake. Due to the need to process barley less extensively than other cereals (as long as the pericarp is broken, consistent and global standards for feeding and processing barley could be feasibly established. In high-starch diets, barley feeding reduces the need for capacious small intestinal starch assimilation, subsequently reducing hindgut starch use and fecal nutrient loss. With its nutritional exclusivities underlined, barley use will be a factual art that can either matchlessly profit or harm rumen microbes, cattle production, farm economics and the environment.

  12. Agrobacterium-mediated transformation of barley (Hordeum vulgare L.).

    Science.gov (United States)

    Ismagul, Ainur; Mazonka, Iryna; Callegari, Corinne; Eliby, Serik

    2014-01-01

    Barley biotechnology requires efficient genetic engineering tools for producing transgenic plants necessary for conducting reverse genetics analyses in breeding and functional genomics research. Agrobacterium-mediated genetic transformation is an important technique for producing barley transgenics with simple low-copy number transgenes. This chapter reports a refined protocol for the systematic high-throughput transformation of the advanced Australian spring barley breeding line WI4330.

  13. Isozyme variation of Microsporum canis and M. cookei from New Zealand.

    Science.gov (United States)

    Simpanya, M F; Jarvis, B D; Baxter, M

    1998-10-01

    Fifty-four isolates of Microsporum canis (Arthroderma otae) from humans, cats and dogs obtained from Auckland, Palmerston North and Wellington, New Zealand and 18 M. cookei and two Diheterospori spp. from soils were examined for variation using eight isozyme loci. M. canis isolates were from infected and non-infected cases. Isozyme analysis separated the three species which were further subdivided into electrophoretic types (ETs). Clustering analysis using normalized percentage disagreement (PTC) average linkage method revealed two clusters for M. cookei with two subclusters in cluster 2. M. canis had three main divisions (clusters 3, 4 and 5) and Diheterospora formed a separate division. The presence of isolates from different sources in the same clusters and lack of statistical significance as measured by confidence intervals suggests the existence of isolates with common lineage.

  14. Protein kinase C isozymes, novel phorbol ester receptors and cancer chemotherapy.

    LENUS (Irish Health Repository)

    Barry, O P

    2012-02-03

    Recent years have seen extensive growth in the understanding of the role(s) of the various PKC isozymes and novel receptors for the phorbol ester tumor promoters. The PKC family of serine-threonine kinases is an important regulator of signaling cascades that control cell proliferation and death, and therefore represent targets for cancer therapy. While past interests have focused on PKC-selective inhibitors, more recently, intensive research has been underway for selective activators and inhibitors for each individual PKC isozyme. In the past few years a large number of PKC activators and inhibitors with potential as anticancer agents have been developed. A number of these compounds are already in Phase II clinical testing. As a new generation of cancer chemotherapeutic agents are designed, developed and put through a series of rigorous clinical trials, we can anticipate achieving exquisite control over PKC-mediated regulatory pathways, leading ultimately to a greater understanding of different cancers.

  15. Intrinsic Peroxidase-like Activity of Ficin

    Science.gov (United States)

    Yang, Yufang; Shen, Dongjun; Long, Yijuan; Xie, Zhixiong; Zheng, Huzhi

    2017-02-01

    Ficin is classified as a sulfhydryl protease isolated from the latex of fig trees. In most cases, a particular enzyme fits a few types of substrate and catalyzes one type of reaction. In this investigation, we found sufficient proofs for the intrinsic peroxidase-like activity of ficin and designed experiments to examine its effectiveness in a variety of scenarios. Ficin can transform peroxidase substrates to colored products in the existence of H2O2. Our results also indicate that the active sites of peroxidase-like activity of ficin are different from that of protease, which reveals that one enzyme may catalyze more than one kind of substrate to perform different types of reactions. On the basis of these findings, H2O2 releasing from MCF-7 cells was detected successfully. Our findings support a wider application of ficin in biochemistry and open up the possibility of utilizing ficin as enzymatic mimics in biotechnology and environmental monitoring.

  16. Changes of Limiting Dextrinase in Germinating Process of Malting Barley

    Institute of Scientific and Technical Information of China (English)

    LIANG Xiu-mei; LI Fen; WANG Hong-zhen; WANG Xing-zhi

    2002-01-01

    Based on five different species of barley, the foot layer analytic method was used to examine the activity and heat-resistance of the limiting dextrinase. The study was conducted on the dynamic changes of several types of the dextrinase in barley germinating process, the effect of temperature on the dextrinase and the divergence of dextrinase in different barley variety. The probability of the dextrinase that as reference index is used for screening and evaluating beer barley was discussed. The importance of dextrinase in brewing and its significant function was also discussed.

  17. Urinary Lactate Dehydrogenase Activity and Its Isozyme Patterns in Kawasaki Disease

    Science.gov (United States)

    Kawamura, Yoichi; Kanai, Takashi; Takizawa, Mari; Yoshida, Yusuke; Tsujita, Yuki; Nonoyama, Shigeaki

    2017-01-01

    Abnormal urinary findings, such as sterile pyuria, proteinuria, and microscopic hematuria, are often seen in the acute phase of Kawasaki disease (KD). We investigated the potential significance of urinary lactate dehydrogenase (U-LDH) activity and its isozyme patterns in KD. Total U-LDH activity and its isozymes (U-LDH1-5) levels were compared among 120 patients with KD, 18 patients with viral infection (VI), and 43 patients with upper urinary tract infection (UTI) and additionally compared between intravenous immunoglobulin (IVIG) responders (n = 89) and nonresponders (n = 31) with KD. Total U-LDH activity was higher in KD (35.4 ± 4.8 IU/L, P < 0.05) and UTI patients (66.0 ± 8.0 IU/L, P < 0.01) than in VI patients (17.0 ± 6.2 IU/L). In the isozyme pattern analysis, KD patients had high levels of U-LDH1 and U-LDH2, while UTI patients had high levels of U-LDH3, U-LDH4, and U-LDH5. Furthermore, IVIG nonresponders of KD had significantly higher levels of total U-LDH activity (45.1 ± 4.7 IU/L, P < 0.05), especially U-LDH1 and U-LDH2 (P < 0.05), than IVIG responders (32.0 ± 2.8 IU/L). KD patients have increased levels of total U-LDH activity, especially U-LDH-1 and U-LDH2, indicating a unique pattern of U-LDH isozymes different from that in UTI patients. PMID:28348604

  18. Urinary Lactate Dehydrogenase Activity and Its Isozyme Patterns in Kawasaki Disease

    Directory of Open Access Journals (Sweden)

    Yoichi Kawamura

    2017-01-01

    Full Text Available Abnormal urinary findings, such as sterile pyuria, proteinuria, and microscopic hematuria, are often seen in the acute phase of Kawasaki disease (KD. We investigated the potential significance of urinary lactate dehydrogenase (U-LDH activity and its isozyme patterns in KD. Total U-LDH activity and its isozymes (U-LDH1-5 levels were compared among 120 patients with KD, 18 patients with viral infection (VI, and 43 patients with upper urinary tract infection (UTI and additionally compared between intravenous immunoglobulin (IVIG responders (n=89 and nonresponders (n=31 with KD. Total U-LDH activity was higher in KD (35.4±4.8 IU/L, P<0.05 and UTI patients (66.0±8.0 IU/L, P<0.01 than in VI patients (17.0±6.2 IU/L. In the isozyme pattern analysis, KD patients had high levels of U-LDH1 and U-LDH2, while UTI patients had high levels of U-LDH3, U-LDH4, and U-LDH5. Furthermore, IVIG nonresponders of KD had significantly higher levels of total U-LDH activity (45.1±4.7 IU/L, P<0.05, especially U-LDH1 and U-LDH2 (P<0.05, than IVIG responders (32.0±2.8 IU/L. KD patients have increased levels of total U-LDH activity, especially U-LDH-1 and U-LDH2, indicating a unique pattern of U-LDH isozymes different from that in UTI patients.

  19. Comparative characterization of pulmonary surfactant aggregates and alkaline phosphatase isozymes in human lung carcinoma tissue.

    Science.gov (United States)

    Iino, Nozomi; Matsunaga, Toshiyuki; Harada, Tsuyoshi; Igarashi, Seiji; Koyama, Iwao; Komoda, Tsugikazu

    2007-05-01

    Alkaline phosphatase (AP) isozymes are surfactant-associated proteins (SPs). Since several different AP isozymes have been detected in the pneumocytes of lung cancer patients, we attempted to identify the relationship between pulmonary surfactant aggregate subtypes and AP isozymes. Pulmonary surfactant aggregates were isolated from carcinoma and non-carcinoma tissues of patients with non-small cell carcinoma of the lung. Upon analysis, ultraheavy, heavy, and light surfactant aggregates were detected in the non-carcinoma tissues, but no ultraheavy surfactant aggregates were found in the carcinoma tissues. Surfactant-associated protein A (SP-A) was detected as two bands (a 27-kDa band and a 54-kDa band) in the ultraheavy, heavy, and light surfactant aggregates found in the non-carcinoma tissues. Although both SP-A bands were detected in the heavy and light surfactant aggregates from adenocarcinoma tissues, the 54-kDa band was not detected in squamous cell carcinoma tissues. Liver AP (LAP) was detected in the heavy and light surfactant aggregates from both non-carcinoma and squamous carcinoma tissues, but not in heavy surfactant aggregates from adenocarcinoma tissues. A larger amount of bone type AP (BAP) was found in light surfactant aggregate fractions from squamous cell carcinomas than those from adenocarcinoma tissues or non-carcinoma tissues from patients with either type of cancer. LAP, BAP, and SP-A were identified immunohistochemically in type II pneumocytes from non-carcinoma tissues and adenocarcinoma cells, but no distinct SP-A staining was observed in squamous cell carcinoma tissues. The present study has thus revealed several differences in pulmonary surfactant aggregates and AP isozymes between adenocarcinoma tissue and squamous cell carcinoma tissue.

  20. Contributions of two cytosolic glutamine synthetase isozymes to ammonium assimilation in Arabidopsis roots.

    Science.gov (United States)

    Konishi, Noriyuki; Ishiyama, Keiki; Beier, Marcel Pascal; Inoue, Eri; Kanno, Keiichi; Yamaya, Tomoyuki; Takahashi, Hideki; Kojima, Soichi

    2016-12-21

    Glutamine synthetase (GS) catalyzes a reaction that incorporates ammonium into glutamate and yields glutamine in the cytosol and chloroplasts. Although the enzymatic characteristics of the GS1 isozymes are well known, their physiological functions in ammonium assimilation and regulation in roots remain unclear. In this study we show evidence that two cytosolic GS1 isozymes (GLN1;2 and GLN1;3) contribute to ammonium assimilation in Arabidopsis roots. Arabidopsis T-DNA insertion lines for GLN1;2 and GLN1;3 (i.e. gln1;2 and gln1;3 single-mutants), the gln1;2:gln1;3 double-mutant, and the wild-type accession (Col-0) were grown in hydroponic culture with variable concentrations of ammonium to compare their growth, and their content of nitrogen, carbon, ammonium, and amino acids. GLN1;2 and GLN1;3 promoter-dependent green fluorescent protein was observed under conditions with or without ammonium supply. Loss of GLN1;2 caused significant suppression of plant growth and glutamine biosynthesis under ammonium-replete conditions. In contrast, loss of GLN1;3 caused slight defects in growth and Gln biosynthesis that were only visible based on a comparison of the gln1;2 single- and gln1;2:gln1;3 double-mutants. GLN1;2, being the most abundantly expressed GS1 isozyme, markedly increased following ammonium supply and its promoter activity was localized at the cortex and epidermis, while GLN1;3 showed only low expression at the pericycle, suggesting their different physiological contributions to ammonium assimilation in roots. The GLN1;2 promoter-deletion analysis identified regulatory sequences required for controlling ammonium-responsive gene expression of GLN1;2 in Arabidopsis roots. These results shed light on GLN1 isozyme-specific regulatory mechanisms in Arabidopsis that allow adaptation to an ammonium-replete environment.

  1. [Genetic control of isozymes in European spruces (Picea abies (L) Karst) of the Ukrainian Carpathian mountains].

    Science.gov (United States)

    Privalikhin, S N; Korshikov, I I; Pirko, N N; Velikorid'ko, T I; Pirko, Ia V

    2006-01-01

    Genetical control of the enzymes GOT, GDH, DIA, MDH, SOD, FDH, ADH, ACP and LAP has been studied in nine natural Carpathian populations of Norway spruce (Picea abies (L.) Karst.) using polyacrylamide gel elecrophoresis and analysis of isozyme variability in 346 trees. Seventy one allel products of 20 gene loci have been clearly established. Segregation analysis of the revealed allele variants confirms their monogenic inheritance.

  2. Shifts in the myosin heavy chain isozymes in the mouse heart result in increased energy efficiency

    Science.gov (United States)

    Hoyer, Kirsten; Krenz, Maike; Robbins, Jeffrey; Ingwall, Joanne S.

    2007-01-01

    Cardiac-specific transgenesis in the mouse is widely used to study the basic biology and chemistry of the heart and to model human cardiovascular disease. A fundamental difference between mouse and human hearts is the background motor protein: mouse hearts contain predominantly the αα-myosin heavy chain (MyHC) isozyme while human hearts contain predominantly the ββ-MyHC isozyme. Although the intrinsic differences in mechanical and enzymatic properties of the αα- and ββ-MyHC molecules are well known, the consequences of isozyme shifts on energetic of the intact beating heart remain unknown. Therefore, we compared the free energy of ATP hydrolysis (|ΔG~ATP|) determined by 31P NMR spectroscopy in isolated perfused littermate mouse hearts containing the same amount of myosin comprised of either >95% αα-MyHC or ~83% ββ-MyHC. |ΔG~ATP| was ~2 kJ mol−1 higher in the ββ-MyHC hearts at all workloads. Furthermore, upon inotropic challenge, hearts containing predominantly ββ-MyHC hearts increased developed pressure more than αα-MyHC hearts whereas heart rate increased more in αα-MyHC hearts. Thus, hearts containing predominantly the ββ-MyHC isozyme are more energy efficient than αα-MyHC hearts. We suggest that these fundamental differences in the motor protein energy efficiency at the whole heart level should be considered when interpreting results using mouse-based cardiovascular modeling of normal and diseased human heart. PMID:17054980

  3. FERTILIZING BREWING BARLEY (Hordeum vulgare L.

    Directory of Open Access Journals (Sweden)

    I. Kádár

    2000-12-01

    Full Text Available Four levels of N, P and K nutrition (poor, moderate, satisfactory and high and all their possible combinations with 64 treatments in two replications (128 plots were studied in a long term field trial on barley yield and malting quality. A standard East-European spring barley "Opal" (bred in Czechoslovakia was grown in 1986, 13th year of the agricultural experiment, involving various crops in previous years, on a calcareous loamy chernozem soil. The optimum fertility levels for yield enhancement resulted in the poorest malting quality: low modification and extract but long saccharification time and high protein. To solve this problem the brewing industry will have to apply the well-known technological methods available since growers are not likely to give up their fertilizers. Applying soil and plant analysis data, having knowledge about both soil and plant optimum values, the danger of the excessive use of fertilizers can be realized and decreased.

  4. Urinary Lactate Dehydrogenase Activity and Its Isozyme Patterns in Kawasaki Disease.

    Science.gov (United States)

    Kawamura, Yoichi; Takeshita, Seiichiro; Kanai, Takashi; Takizawa, Mari; Yoshida, Yusuke; Tsujita, Yuki; Nonoyama, Shigeaki

    2017-01-01

    Abnormal urinary findings, such as sterile pyuria, proteinuria, and microscopic hematuria, are often seen in the acute phase of Kawasaki disease (KD). We investigated the potential significance of urinary lactate dehydrogenase (U-LDH) activity and its isozyme patterns in KD. Total U-LDH activity and its isozymes (U-LDH1-5) levels were compared among 120 patients with KD, 18 patients with viral infection (VI), and 43 patients with upper urinary tract infection (UTI) and additionally compared between intravenous immunoglobulin (IVIG) responders (n = 89) and nonresponders (n = 31) with KD. Total U-LDH activity was higher in KD (35.4 ± 4.8 IU/L, P IVIG nonresponders of KD had significantly higher levels of total U-LDH activity (45.1 ± 4.7 IU/L, P IVIG responders (32.0 ± 2.8 IU/L). KD patients have increased levels of total U-LDH activity, especially U-LDH-1 and U-LDH2, indicating a unique pattern of U-LDH isozymes different from that in UTI patients.

  5. Effects of Pristane on Cytochrome P450 Isozyme Expression in Rat Tissues

    Directory of Open Access Journals (Sweden)

    Marvin A. Cuchens

    2005-04-01

    Full Text Available Chemical carcinogenesis studies are powerful tools to obtain information on potential mechanisms of chemical factors for malignancies. In this study Western blot analyses, using monoclonal antibodies specific for three different cytochrome P450 (CYP isozymes (CYP1A1, CYP1A2 and CYP2B, were employed to examine the effect(s of 3-methylcholanthrene and/or pristane (2,6,10,14-tetramethylpentadecane on the basal and inducible levels of expression of CYP proteins within Copenhagen rat tissues. Pristane exposure led to tissue specific differences in the CYP isozymes expressed and elicited increased CYP protein expression over 3-methylcholanthrene induced levels in microsomes isolated from liver, Peyer's Patches, and thymus. Within the context of the chemical carcinogenesis model employed in this study, these observations correlated with the induction of B-cell malignancies by low doses of 3-methylcholanthrene and of thymic lymphomas by a high 3-methylcholanthrene dose. The data suggest that pristane treatment affects CYP isozyme expression. This pristane-mediated effect clearly could be a contributing factor in the chemical carcinogenesis of the previously observed lymphoid malignancies, and a possible basis for the tumor enhancing effects of pristane.

  6. Effect of biliary obstruction and internal biliary drainage on hepatic cytochrome P450 isozymes in rats

    Institute of Scientific and Technical Information of China (English)

    Shintaro Fukushima; Hiroyasu Okuno; Nobuyuki Shibatani; Yoshitsugu Nakahashi; Toshihito Seki; Kazuichi Okazaki

    2008-01-01

    AIM: To investigate the total cytochrome P450 (CYP)content, microsomal mixed-function oxidase (MFO)activity, and expression of mRNAs for various CYP isozymes in a simple rat model of reversible obstructive jaundice.METHODS: Obstructive jaundice was created in male rats by causing bile duct obstruction with polyester tape.In another group of rats, bile duct obstruction was followed by internal biliary drainage after releasing the tape.The expression of various CYP isozyme mRNAs was semi-quantitatively assessed by competitive RTPCR.RESULTS: The total CYP content and microsomal MFO activity showed a significant decrease after biliary obstruction, but returned to respective control levels after biliary drainage.A marked reduction in the expression of CYPIA2, 2B1/2, 2Cll, 2E1, 3A1, and 3A2 mRNA was detected during biliary obstruction,while expression increased significantly toward the control level after biliary drainage.Although expression of CYP4A1 mRNA showed no reduction during biliary obstruction, it still increased significantly after biliary drainage.CONCLUSION: These results suggest that not only obstructive jaundice, but also the subsequent internal biliary drainage may affect regulatory medications of the synthesis of individual CYP isozymes differently.

  7. Tissue expression and stock variation of isozymes of stone flounder ( Kareius bicoloratus)

    Science.gov (United States)

    Xu, Jianpeng; Zhang, Quanqi; Qi, Jie; Wang, Zhigang

    2007-04-01

    Tissue expression and stock variation of isozymes of stone flounder ( Kareius bicoloratus) were analyzed with horizontal starch gel electrophoresis. For the fourteen enzymes assayed, 31 loci were recorded. The results indicated that all the isozymes examined were obviously tissue-specific. The expressions of SOD *, GDH *, G3PDH-2 * and ADH-2 * were detected only in liver, SDH-1 *, MDH-1 * and ADH-1 * only in muscle, and LDH-B * and LDH-C * only in eyes. In comparison, MDH-2 *, GPI-3 * and SDH-2 * were detected in all tissues examined. Other loci examined were detected in a variety of tissues. Muscle and liver were selected to detect the isozyme variation of the two geographic stocks of Qingdao and Weihai, Shandong Province, China. The percentages of polymorphic loci ( P 0.99) were 29.17% and 25.00%, the observed heterozygosities ( H 0) were 0.028 ± 0.014 and 0.040 ± 0.019, and the expected heterozygosities ( He) were 0.039 ± 0.017 and 0.052 ± 0.022 in Qingdao and Weihai stock, respectively. The coefficient of gene differentiation ( F st) and genetic distance ( D) between the two stocks was 0.012 and 0.0011, respectively, indicating that the genetic differentiation is low between them. Compared with other species of Pleuronectiformes, both the percentage of polymorphic loci and the mean heterozygosity of K. bicoloratus were at a middle level.

  8. Allosteric regulation of pyruvate kinase M2 isozyme involves a cysteine residue in the intersubunit contact.

    Science.gov (United States)

    Ikeda, Y; Noguchi, T

    1998-05-15

    Pyruvate kinase M2 isozyme mutants with amino acid substitutions in the subunit interface were prepared and characterized. The substitutions were made in the allosteric M2 isozyme by the corresponding residues of the nonallosteric M1 isozyme to identify the residue involved in the allosteric effects. The replacement of Cys-423 by Leu led to substantial loss of both homotropic and heterotropic allosteric effects while the substitutions at Phe-389, Arg-398, Ala-401, Pro-402, Thr-408, and Ile-427 did not. The altered kinetic properties of the Cys-423-substituted mutant resulted from the shift of the allosteric transition toward the active R-state since the mutant exhibits the allosteric properties in the presence of an allosteric inhibitor, L-phenylalanine. The inverse correlation between the hydrophobicity of residue 423 and the extent of stabilization of the R-state was found by analysis of mutants with un-ionizable amino acids at position 423. Furthermore, the modification of Cys-423 with methyl methanethiosulfonate led to a shift of the allosteric transition toward the R-state, probably the result of increased hydrophobicity of the residue. These results suggest that Cys-423 is involved in the allosteric regulation of the enzyme through hydrophobic interactions.

  9. Isozyme and RAPD studies in Prosopis glandulosa and P. Velutina (Leguminosae, Mimosoideae

    Directory of Open Access Journals (Sweden)

    Bessega Cecilia

    2000-01-01

    Full Text Available Allozyme and random amplified polymorphic DNA (RAPD techniques have been compared for their usefulness for genetic and taxonomic studies in Prosopis glandulosa and P. velutina populations. Isozymes and RAPDs yielded similarly high estimates of genetic variability. Genetic structure and differentiation were analyzed through non-hierarchical Wright's F DT. For all populations considered, both markers produced low gene flow (Nm 1, in agreement with that expected for conspecific populations. However, in RAPD data the expected reduction in F DT and the increase in Nm were not observed. Correlation between F DT and geographical distance matrices (Mantel test for all populations was significant (P = 0.02 when based on isozymes, but not so (P = 0.33 when based on RAPDs. No significant associations among genetic and geographical or climatic variables were observed. Two isoenzyme systems (GOT and PRX enabled us to distinguish between P. glandulosa and P. velutina, but no diagnostic band for recognition of populations or species studied here were detected by RAPD. However, RAPD markers showed higher values for genetic differentiation among conspecific populations of P. glandulosa and a lower coefficient of variation than those obtained from isozymes.

  10. Taxonomy Icon Data: barley [Taxonomy Icon

    Lifescience Database Archive (English)

    Full Text Available barley Hordeum vulgare Hordeum_vulgare_L.png Hordeum_vulgare_NL.png Hordeum_vulgare_S.png Hordeum_vu...lgare_NS.png http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Hordeum+vulgare&t=L http://bi...osciencedbc.jp/taxonomy_icon/icon.cgi?i=Hordeum+vulgare&t=NL http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Hordeum+vu...lgare&t=S http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Hordeum+vulgare&t=NS ...

  11. Search for endophytic diazotrophs in barley seeds

    Directory of Open Access Journals (Sweden)

    Myriam S. Zawoznik

    2014-06-01

    Full Text Available Eight endophytic isolates assigned to Pseudomonas, Azospirillum, and Bacillus genera according to pheno-genotypic features were retrieved from barley seeds under selective pressure for nitrogen-fixers. Genetic relationships among related isolates were investigated through RAPD. Six isolates displayed nitrogen-fixing ability, while all could biosynthesize indolacetic acid in vitro and showed no antibiosis effects against Azospirillum brasilense Az39, a recognized PGPR.

  12. Transgenic Wheat, Barley and Oats: Future Prospects

    Science.gov (United States)

    Dunwell, Jim M.

    Following the success of transgenic maize and rice, methods have now been developed for the efficient introduction of genes into wheat, barley and oats. This review summarizes the present position in relation to these three species, and also uses information from field trial databases and the patent literature to assess the future trends in the exploitation of transgenic material. This analysis includes agronomic traits and also discusses opportunities in expanding areas such as biofuels and biopharming.

  13. Search for endophytic diazotrophs in barley seeds.

    Science.gov (United States)

    Zawoznik, Myriam S; Vázquez, Susana C; Díaz Herrera, Silvana M; Groppa, María D

    2014-01-01

    Eight endophytic isolates assigned to Pseudomonas, Azospirillum, and Bacillus genera according to pheno-genotypic features were retrieved from barley seeds under selective pressure for nitrogen-fixers. Genetic relationships among related isolates were investigated through RAPD. Six isolates displayed nitrogen-fixing ability, while all could biosynthesize indolacetic acid in vitro and showed no antibiosis effects against Azospirillum brasilense Az39, a recognized PGPR.

  14. Functional Analysis of Barley Powdery Mildew Effector Candidates and Identification of their Barley Targets

    DEFF Research Database (Denmark)

    Ahmed, Ali Abdurehim

    about the function of many CSEPs in virulence and the identities of their host targets. In this PhD study, we investigated the function of nine CSEPs and found that CSEP0081, CSEP0105, CSEP0162 and CSEP0254 act as effectors by promoting the Bgh infection success. Independent silencing of these CSEPs...... to the cytosol and the nucleus of barley epidermal cells. Furthermore, CSEP0162 and CSEP0254 accumulated in the extrahaustorial matrix in Bgh-infected cells. This implies that their virulence targets may localize in the same cellular compartments. Using yeast two-hybrid screens, two barley small heat shock...... misfolding and aggregation. Through their chaperone activity, some sHsps contribute to pathogen defence by stabilizing intracellular proteins, including resistance and defence signalling proteins. In this study, we validated the chaperone activity of the barley Hsp16.9, which prevented the aggregation...

  15. Stability of Barley stripe mosaic virus induced gene silencing in barley

    DEFF Research Database (Denmark)

    Bruun-Rasmussen, Marianne; Madsen, Christian Toft; Jessing, Stine

    2007-01-01

    Virus-induced gene silencing (VIGS) can be used as a powerful tool for functional genomics studies in plants. With this approach, it is possible to target most genes and downregulate the messenger (m)RNA in a sequence-specific manner. Barley stripe mosaic virus (BSMV) is an established VIGS vector...... for barley and wheat; however, silencing using this vector is generally transient, with efficient silencing often being confined to the first two or three systemically infected leaves. To investigate this further, part of the barley Phytoene desaturase (PDS) gene was inserted into BSMV and the resulting...... inoculation, although large parts of the insert had been lost from the virus vector. The instability of the insert, observed consistently throughout our experiments, offers an explanation for the transient nature of silencing when using BSMV as a VIGS vector....

  16. Peroxidase complex with concomitant anodal and cathodal variation in red-fruited tomato species.

    Science.gov (United States)

    Rick, C M; Fobes, J F

    1976-03-01

    Four groups of bands (a-d) are controlled by 19 alleles of the Peroxidase-4 (Prx-4) complex in the red-fruited tomato species, Lycopersicon esculentum and L. pimpinellifolium. Heterozygotes can be detected by virtue of codominance in all combinations except a few in which bands of single groups are absent ("semi-null" alleles). No recombinations were detected in 7419 F(2) segregants of 53 different combinations of alleles. A maximum fiducial limit (P = 0.01) of 0.08% crossing-over between any Prx-4 band groups is estimated. Variation of the anodal b bands is absolutely associated with that of the cathodal d band in respect to presence versus absence and direction of migration. In respect to the origin of these variants, the probability of 18 instances of simultaneous mutation of genes at two loci, always in such complete agreement, is so remote that no more than one locus could conceivably govern b and d. The disposition of a is not similarly associated with that of the other bands, while that of the faint-staining c could not always be reliably resolved. The negation of all save extremely low recombination rates and the observed concomitant variation of b and d strongly support the concept of single locus control of all Prx-4 banding, this hypothesis being espoused until rejection should be required be required by future research. Models of single locus control of several isozymes are discussed.

  17. The NAC transcription factors of barley

    DEFF Research Database (Denmark)

    Wagner, Michael; Holm, Preben Bach; Gregersen, Per L.

    2011-01-01

    ). From these data we have identified not only putative regulators of leaf senescence (HvNAC005, HvNAC027 and HvNAC029), but also possible regulators of secondary wall synthesis (HvNAC033, HvNAC034 and HvNAC039), lateral root formation (HvNAC022) and seed development (HvNAC017, HvNAC018, HvNAC019 and Hv...... genes characterized so far have regulatory functions in a broad range of plant developmental processes and tolerances to both biotic and abiotic stresses. This makes the NAC family highly interesting target genes for plant researchers and breeders. As part of a larger project on the identification...... of Hordeum vulgare (barley) leaf senescence regulators, we have attempted to characterize for the first time all presently available barley NAC genes (HvNACs). By searching the NCBI barley EST database using the tBLASTn function, with all known NAC genes from Brachypodium and rice as input, in combination...

  18. Peroxidase-benzhydroxamic acid complexes: spectroscopic evidence that a Fe-H2O distance of 2.6 A can correspond to hexa-coordinate high-spin heme.

    Science.gov (United States)

    Smulevich, G; Feis, A; Indiani, C; Becucci, M; Marzocchi, M P

    1999-02-01

    Resonance Raman (RR) spectra have been obtained for single-crystal horseradish peroxidase isozyme C complexed with benzhydroxamic acid (BHA). The data are compared with those obtained in solution by both RR and electronic absorption spectroscopies at room and low (12-80 K) temperatures. Moreover, the analysis has been extended to Coprinus cinereus peroxidase complexed with BHA. The results obtained for the two complexes are very similar and are consistent with the presence of an aqua six-coordinate high-spin heme. Therefore it can be concluded that despite the rather long Fe-H2O distance of 2.6-2.7 A found by X-ray crystallography in both complexes, the distal water molecule can still coordinate to the heme iron.

  19. Characterization of aldehyde dehydrogenase isozymes in ovarian cancer tissues and sphere cultures

    Directory of Open Access Journals (Sweden)

    Saw Yu-Ting

    2012-08-01

    Full Text Available Abstract Background Aldehyde dehydrogenases belong to a superfamily of detoxifying enzymes that protect cells from carcinogenic aldehydes. Of the superfamily, ALDH1A1 has gained most attention because current studies have shown that its expression is associated with human cancer stem cells. However, ALDH1A1 is only one of the 19 human ALDH subfamilies currently known. The purpose of the present study was to determine if the expression and activities of other major ALDH isozymes are associated with human ovarian cancer and ovarian cancer sphere cultures. Methods Immunohistochemistry was used to delineate ALDH isozyme localization in clinical ovarian tissues. Western Blot analyses were performed on lysates prepared from cancer cell lines and ovarian cancer spheres to confirm the immunohistochemistry findings. Quantitative reverse transcription-polymerase chain reactions were used to measure the mRNA expression levels. The Aldefluor® assay was used to measure ALDH activity in cancer cells from the four tumor subtypes. Results Immunohistochemical staining showed significant overexpression of ALDH1A3, ALDH3A2, and ALDH7A1 isozymes in ovarian tumors relative to normal ovarian tissues. The expression and activity of ALDH1A1 is tumor type-dependent, as seen from immunohistochemisty, Western blot analysis, and the Aldefluor® assay. The expression was elevated in the mucinous and endometrioid ovarian epithelial tumors than in serous and clear cell tumors. In some serous and most clear cell tumors, ALDH1A1 expression was found in the stromal fibroblasts. RNA expression of all studied ALDH isozymes also showed higher expression in endometrioid and mucinous tumors than in the serous and clear cell subtypes. The expression of ALDH enzymes showed tumor type-dependent induction in ovarian cancer cells growing as sphere suspensions in serum-free medium. Conclusions The results of our study indicate that ALDH enzyme expression and activity may be associated

  20. Variation of morphology, isozymic and vitamin C content of dragon fruit varieties

    Directory of Open Access Journals (Sweden)

    EDWI MAHAJOENO

    2009-11-01

    Full Text Available Rahmawati B, Mahajoeno E. 2009. Variation of morphology, isozymic and vitamin C content of dragon fruit varieties. Nusantara Bioscience 1: 131-137. The aims of the research was to study the variation of morphology, the band pattern of isozyme, and vitamin C content of dragon fruit (Hylocereus spp. varieties such as super red, red and white from Pasuruan (East Java, Sukoharjo and Klaten (Central Java, and Bantul districts (Yogyakarta. Morphological character were carried include fruit, stem, and flowers of each variety of dragon fruit. The isozymic pattern was analyzed using NTSYS 2.02i. The data matrix was counted based on the DICE coefficient. The clustering was done by applying UPGMA which counted through SHAN. Vitamin C content measured by titration method then analyzed descriptively. The results showed that the higher vitamin C content was found from super red of Pasuruan (6.00 and then followed by red color (5.376 and super red (5.113 both from Bantul. The morphological variation on the stem and petal colors, and fruits were also shown by the isozymic data of three varieties of dragon fruits collected from four separated locations. Esterase (EST showed 18 bands and forming four (4 groups based on 75% genetic similarity index. The specific band occurred on Rf 0.633 of red varieties of dragon fruit from Bantul and on Rf 0.755 from Pasuruan. The specific band also occurs on Rf 0.347 of white variety from Bantul and on Rf 0.510 and on Rf 0.633 from Klaten. Glutamic oxaloacetic transaminase (GOT enzyme shows 12 bands and also forming four groups with a little difference for member in the fourth group. The specific band occurs on Rf 0.321 of red color fruit from Pasuruan. The specific band also occurs on the white from Pasuruan on Rf 0.446 and on Rf 0.482. The variation of dragon fruits were also supported by isozymic data indicated that the morphological character were in accordance with the genetics data.

  1. Widespread occurrence of expressed fungal secretory peroxidases in forest soils.

    Science.gov (United States)

    Kellner, Harald; Luis, Patricia; Pecyna, Marek J; Barbi, Florian; Kapturska, Danuta; Krüger, Dirk; Zak, Donald R; Marmeisse, Roland; Vandenbol, Micheline; Hofrichter, Martin

    2014-01-01

    Fungal secretory peroxidases mediate fundamental ecological functions in the conversion and degradation of plant biomass. Many of these enzymes have strong oxidizing activities towards aromatic compounds and are involved in the degradation of plant cell wall (lignin) and humus. They comprise three major groups: class II peroxidases (including lignin peroxidase, manganese peroxidase, versatile peroxidase and generic peroxidase), dye-decolorizing peroxidases, and heme-thiolate peroxidases (e.g. unspecific/aromatic peroxygenase, chloroperoxidase). Here, we have repeatedly observed a widespread expression of all major peroxidase groups in leaf and needle litter across a range of forest ecosystems (e.g. Fagus, Picea, Acer, Quercus, and Populus spp.), which are widespread in Europe and North America. Manganese peroxidases and unspecific peroxygenases were found expressed in all nine investigated forest sites, and dye-decolorizing peroxidases were observed in five of the nine sites, thereby indicating biological significance of these enzymes for fungal physiology and ecosystem processes. Transcripts of selected secretory peroxidase genes were also analyzed in pure cultures of several litter-decomposing species and other fungi. Using this information, we were able to match, in environmental litter samples, two manganese peroxidase sequences to Mycena galopus and Mycena epipterygia and one unspecific peroxygenase transcript to Mycena galopus, suggesting an important role of this litter- and coarse woody debris-dwelling genus in the disintegration and transformation of litter aromatics and organic matter formation.

  2. Occurrence and properties of petunia peroxidase a.

    NARCIS (Netherlands)

    Hendriks, Th.

    1989-01-01

    Peroxidases are probably the most extensively studied enzymes in higher plants. Various isoenzymes occur as soluble proteins in the apoplast and in the vacuole, or are bound to membranes and cell walls. Their occurrence is often organ-specific and developmentally controlled, and there is circumstant

  3. Occurrence and properties of Petunia peroxidase a

    NARCIS (Netherlands)

    Hendriks, T.

    1989-01-01

    Peroxidases are probably the most extensively studied enzymes in higher plants. Various isoenzymes occur as soluble proteins in the apoplast and in the vacuole, or are bound to membranes and cell walls. Their occurrence is often organ-specific and developmentally controlled, and there is

  4. Inhibition of Heme Peroxidases by Melamine

    Directory of Open Access Journals (Sweden)

    Pattaraporn Vanachayangkul

    2012-01-01

    Full Text Available In 2008 melamine-contaminated infant formula and dairy products in China led to over 50,000 hospitalizations of children due to renal injuries. In North America during 2007 and in Asia during 2004, melamine-contaminated pet food products resulted in numerous pet deaths due to renal failure. Animal studies have confirmed the potent renal toxicity of melamine combined with cyanuric acid. We showed previously that the solubility of melamine cyanurate is low at physiologic pH and ionic strength, provoking us to speculate how toxic levels of these compounds could be transported through the circulation without crystallizing until passing into the renal filtrate. We hypothesized that melamine might be sequestered by heme proteins, which could interfere with heme enzyme activity. Four heme peroxidase enzymes were selected for study: horseradish peroxidase (HRP, lactoperoxidase (LPO, and cyclooxygenase-1 and -2 (COX-1 and -2. Melamine exhibited noncompetitive inhibition of HRP (9.5±0.7mM, and LPO showed a mixed model of inhibition (14.5±4.7mM. The inhibition of HRP and LPO was confirmed using a chemiluminescent peroxidase assay. Melamine also exhibited COX-1 inhibition, but inhibition of COX-2 was not detected. Thus, our results demonstrate that melamine inhibits the activity of three heme peroxidases.

  5. Occurrence and properties of Petunia peroxidase a

    NARCIS (Netherlands)

    Hendriks, T.

    1989-01-01

    Peroxidases are probably the most extensively studied enzymes in higher plants. Various isoenzymes occur as soluble proteins in the apoplast and in the vacuole, or are bound to membranes and cell walls. Their occurrence is often organ-specific and developmentally controlled, and there is ci

  6. Guaiacol Peroxidase Zymography for the Undergraduate Laboratory

    Science.gov (United States)

    Wilkesman, Jeff; Castro, Diana; Contreras, Lellys M.; Kurz, Liliana

    2014-01-01

    This laboratory exercise presents a novel way to introduce undergraduate students to the specific detection of enzymatic activity by electrophoresis. First, students prepare a crude peroxidase extract and then analyze the homogenate via electrophoresis. Zymography, that is, a SDS-PAGE method to detect enzyme activity, is used to specifically…

  7. Bioconjugation of antibodies to horseradish peroxidase (hrp)

    Science.gov (United States)

    The bioconjugation of an antibody to an enzymatic reporter such as horseradish peroxidase (HRP) affords an effective mechanism by which immunoassay detection of a target antigen can be achieved. The use of heterobifunctional cross—linkers to covalently link antibodies to HRP provides a simple and c...

  8. Heterologous Expression of Peroxidases : Chapter 12

    NARCIS (Netherlands)

    Lokman, Christien; Weert, S. de

    2010-01-01

    This monograph describes many applications of peroxidase-based biocatalysis in the biotechnology industry. The need for such a book emerges from the considerable amount of new data regarding the phylogeny, reaction mechanisms, thermodynamic characterization and structural features of fungal and plan

  9. Calnexin overexpression increases manganese peroxidase production in Aspergillus niger

    NARCIS (Netherlands)

    Conesa, A.; Jeenes, D.; Archer, D.B.; Hondel, C.A.M.J.J. van den; Punt, P.J.

    2002-01-01

    Heme-containing peroxidases from white rot basidiomycetes, in contrast to most proteins of fungal origin, are poorly produced in industrial filamentous fungal strains. Factors limiting peroxidase production are believed to operate at the posttranslational level. In particular, insufficient

  10. Calnexin overexpression increases manganese peroxidase production in Aspergillus niger

    NARCIS (Netherlands)

    Conesa, A.; Jeenes, D.; Archer, D.B.; Hondel, C.A.M.J.J. van den; Punt, P.J.

    2002-01-01

    Heme-containing peroxidases from white rot basidiomycetes, in contrast to most proteins of fungal origin, are poorly produced in industrial filamentous fungal strains. Factors limiting peroxidase production are believed to operate at the posttranslational level. In particular, insufficient availabil

  11. 7 CFR 457.118 - Malting barley crop insurance.

    Science.gov (United States)

    2010-01-01

    ... Barley Price and Quality Endorsement (This is a continuous endorsement. Refer to section 2 of the Common... all quality criteria contained herein or grades U.S. No. 4 or lower in accordance with the grades and... coverage for malting barley production and quality losses at a price per bushel greater than that offered...

  12. Barley metallothioneins differ in ontogenetic pattern and response to metals

    DEFF Research Database (Denmark)

    Schiller, Michaela; Hegelund, Josefine Nymark; Pedas, Pai

    2014-01-01

    The barley genome encodes a family of 10 metallothioneins (MTs) that have not previously been subject to extensive gene expression profiling. We show here that expression of MT1a, MT2b1, MT2b2 and MT3 in barley leaves increased more than 50-fold during the first 10 d after germination. Concurrent...

  13. Analysis of Pregerminated Barley Using Hyperspectral Image Analysis

    DEFF Research Database (Denmark)

    Arngren, Morten; Hansen, Per Waaben; Eriksen, Birger

    2011-01-01

    Pregermination is one of many serious degradations to barley when used for malting. A pregerminated barley kernel can under certain conditions not regerminate and is reduced to animal feed of lower quality. Identifying pregermination at an early stage is therefore essential in order to segregate ...

  14. Progressive hull removal from barley using the Fitzpatrick comminuting mill

    Science.gov (United States)

    The objective of the study was to explore an alternative use of the Fitzpatrick Comminuting Machine: to use it to remove the hull from hulled barley while keeping the barley kernel intact. Traditionally, this mill is used to grind material, but we have recently discovered that it also has the abili...

  15. Combining unmalted barley and pearling gives good quality brewing

    NARCIS (Netherlands)

    Donkelaar, van Laura H.G.; Hageman, Jos A.; Oguz, Serhat; Noordman, Tom R.; Boom, Remko M.; Goot, van der Atze Jan

    2016-01-01

    Brewing with unmalted barley can reduce the use of raw materials, thereby increasing the efficiency of the brewing process. However, unmalted barley contains several undesired components for brewing and has a low enzymatic activity. Pearling, an abrasive milling method, has been proposed as a pre

  16. Low Phytic Acid Barley Responses to Phosphorus Rates

    Science.gov (United States)

    Low phytic acid (LPA) barley (Hordeum vulgare L.) cultivars partition phosphorus in seed tissue differently than conventional barley cultivars through a reduction in seed phytic acid (myo-inositol-1,2,3,4,5,6-hexkisphosphate) coupled with an increase in inorganic phosphorus. The response of the LPA...

  17. Feruloylated arabinoxylans are oxidatively cross-linked by extracellular maize peroxidase but not by horseradish peroxidase.

    Science.gov (United States)

    Burr, Sally J; Fry, Stephen C

    2009-09-01

    Covalent cross-linking of soluble extracellular arabinoxylans in living maize cultures, which models the cross-linking of wall-bound arabinoxylans, is due to oxidation of feruloyl esters to oligoferuloyl esters and ethers. The oxidizing system responsible could be H2O2/peroxidase, O2/laccase, or reactive oxygen species acting non-enzymically. To distinguish these possibilities, we studied arabinoxylan cross-linking in vivo and in vitro. In living cultures, exogenous, soluble, extracellular, feruloylated [pentosyl-3H]arabinoxylans underwent cross-linking, beginning abruptly 8 d after sub-culture. Cross-linking was suppressed by iodide, an H2O2 scavenger, indicating dependence on endogenous H2O2. However, exogenous H2O2 did not cause precocious cross-linking, despite the constant presence of endogenous peroxidases, suggesting that younger cultures contained natural cross-linking inhibitors. Dialysed culture-filtrates cross-linked [3H]arabinoxylans in vitro only if H2O2 was also added, indicating a peroxidase requirement. This cross-linking was highly ionic-strength-dependent. The peroxidases responsible were heat-labile, although relatively heat-stable peroxidases (assayed on o-dianisidine) were also present. Surprisingly, added horseradish peroxidase, even after heat-denaturation, blocked the arabinoxylan-cross-linking action of maize peroxidases, suggesting that the horseradish protein was a competing substrate for [3H]arabinoxylan coupling. In conclusion, we show for the first time that cross-linking of extracellular arabinoxylan in living maize cultures is an action of apoplastic peroxidases, some of whose unusual properties we report.

  18. Evidence for thiocyanate-sensitive peroxidase activity in human saliva.

    OpenAIRE

    Cowman, R A; Baron, S S; Obenauf, S D; Byrnes, J J

    1983-01-01

    A procedure was developed for determining the relative levels of lactoperoxidase, leukocyte myeloperoxidase, and thiocyanate-sensitive peroxidase in human saliva. With this procedure, most of the peroxidase activity in whole saliva from normal (those without cancer) subjects was found to be associated with lactoperoxidase and thiocyanate-sensitive peroxidase, with only a minor contribution from leukocyte myeloperoxidase. In contrast, thiocyanate-sensitive peroxidase and leukocyte myeloperoxid...

  19. Barley Stripe Mosaic Virus and the Frequency of Triploids and Aneuploids in Barley

    DEFF Research Database (Denmark)

    Sandfær, J.

    1973-01-01

    BSMV infection caused a pronounced increase in the frequency of triploid and aneuploid seeds in eleven barley varieties, but with considerable variation in frequency among varieties. In some of the varieties triploids exceeded three per cent. In virus-free material a few triploids were found in m...

  20. Giemsa C-banding of Barley Chromosomes. IV. Chromosomal Constitution of Autotetraploid Barley

    DEFF Research Database (Denmark)

    Linde-Laursen, Ib

    1984-01-01

    The progeny of an autotetraploid barley plant (C1) consisted of 45 tetraploids and 33 aneuploids. Giemsa C-banding was used to identify each of the chromosomes in 20 euploid and 31 aneuploid C2--seedlings, and in 11 C3--offspring of aneuploid C2--plants. The euploid C2--seedlings all had four hom...

  1. Studies on the production of fungal peroxidases in Aspergillus niger

    NARCIS (Netherlands)

    Conesa, A.; Hondel, C.A.M.J.J. van den; Punt, P.J.

    2000-01-01

    To get insight into the limiting factors existing for the efficient production of fungal peroxidase in filamentous fungi, the expression of the Phanerochaete chrysosporium lignin peroxidase H8 (lipA) and manganese peroxidase (MnP) H4 (mnp1) genes in Aspergillus niger has been studied. For this

  2. Studies on the production of fungal peroxidases in Aspergillus niger

    NARCIS (Netherlands)

    Conesa, A.; Hondel, C.A.M.J.J. van den; Punt, P.J.

    2000-01-01

    To get insight into the limiting factors existing for the efficient production of fungal peroxidase in filamentous fungi, the expression of the Phanerochaete chrysosporium lignin peroxidase H8 (lipA) and manganese peroxidase (MnP) H4 (mnp1) genes in Aspergillus niger has been studied. For this purpo

  3. Studies on the production of fungal peroxidases in Aspergillus niger

    NARCIS (Netherlands)

    Conesa, A.; Hondel, C.A.M.J.J. van den; Punt, P.J.

    2000-01-01

    To get insight into the limiting factors existing for the efficient production of fungal peroxidase in filamentous fungi, the expression of the Phanerochaete chrysosporium lignin peroxidase H8 (lipA) and manganese peroxidase (MnP) H4 (mnp1) genes in Aspergillus niger has been studied. For this purpo

  4. [Characterization of lignin and Mn peroxidases from Phanerochaete chrysosporium

    Energy Technology Data Exchange (ETDEWEB)

    1992-01-01

    Lignin peroxidases were investigated with respect to enzyme kinetics and NMR spectroscopy of the heme domain. MN peroxidases were studied with respect to the role of oxalate in enzyme activity, the NMR spectroscopy of the heme domain. Gene expression of both lignin and MN peroxidases were examined as well as expression of site-directed mutants aimed at scale up production of these enzymes.

  5. Saline-Dependent Regulation of Manganese Peroxidase Genes in the Hypersaline-Tolerant White Rot Fungus Phlebia sp. Strain MG-60▿

    Science.gov (United States)

    Kamei, Ichiro; Daikoku, Chieko; Tsutsumi, Yuji; Kondo, Ryuichiro

    2008-01-01

    The expression pattern of manganese peroxidases (MnPs) in nitrogen-limited cultures of the saline-tolerant fungus Phlebia sp. strain MG-60 is differentially regulated under hypersaline conditions at the mRNA level. When MG-60 was cultured in nitrogen-limited medium (LNM) containing 3% (wt/vol) sea salts (LN-SSM), higher activity of MnPs was observed than that observed in normal medium (LNM). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis demonstrated that two MnP isoenzymes were de novo synthesized in the culture of LN-SSM. Three MnP-encoding genes (MGmnp1, MGmnp2, and MGmnp3) were isolated by reverse transcription (RT)-PCR and rapid amplification of cDNA ends PCR techniques. The corresponding isozymes were identified by peptide mass fingerprinting analysis. MnP isozymes encoded by MGmnp2 and MGmnp3 were observed mainly in LN-SSM. Real-time RT-PCR analysis revealed high levels of MGmnp2 and MGmnp3 transcripts in LN-SSM 48 h after the addition of 2% NaCl. The induction of MnP production and the accumulation of gene transcripts by saline were well correlated in the presence of Mn2+. However, in the absence of Mn2+, there was no clear correlation between mnp transcripts levels and MnP activity, suggesting posttranscriptional regulation by Mn2+. PMID:18310430

  6. Saline-dependent regulation of manganese peroxidase genes in the hypersaline-tolerant white rot fungus Phlebia sp. strain MG-60.

    Science.gov (United States)

    Kamei, Ichiro; Daikoku, Chieko; Tsutsumi, Yuji; Kondo, Ryuichiro

    2008-05-01

    The expression pattern of manganese peroxidases (MnPs) in nitrogen-limited cultures of the saline-tolerant fungus Phlebia sp. strain MG-60 is differentially regulated under hypersaline conditions at the mRNA level. When MG-60 was cultured in nitrogen-limited medium (LNM) containing 3% (wt/vol) sea salts (LN-SSM), higher activity of MnPs was observed than that observed in normal medium (LNM). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis demonstrated that two MnP isoenzymes were de novo synthesized in the culture of LN-SSM. Three MnP-encoding genes (MGmnp1, MGmnp2, and MGmnp3) were isolated by reverse transcription (RT)-PCR and rapid amplification of cDNA ends PCR techniques. The corresponding isozymes were identified by peptide mass fingerprinting analysis. MnP isozymes encoded by MGmnp2 and MGmnp3 were observed mainly in LN-SSM. Real-time RT-PCR analysis revealed high levels of MGmnp2 and MGmnp3 transcripts in LN-SSM 48 h after the addition of 2% NaCl. The induction of MnP production and the accumulation of gene transcripts by saline were well correlated in the presence of Mn(2+). However, in the absence of Mn(2+), there was no clear correlation between mnp transcripts levels and MnP activity, suggesting posttranscriptional regulation by Mn(2+).

  7. Pleurotus ostreatus, an edible mushroom, enhances glucose 6-phosphate dehydrogenase, ascorbate peroxidase and reduces xanthine dehydrogenase in major organs of aged rats.

    Science.gov (United States)

    Thomas, Philip Aloysius; Geraldine, Pitchairaj; Jayakumar, Thanasekaran

    2014-05-01

    Aging is now considered to be associated with an elevation in oxidative damage to macromolecules and enhanced levels of inflammation. Therefore, inhibition of age-related oxidative stress by natural supplement is an important study. To investigate whether the treatment with Pleurotus ostreatus (Jacq.: Fr) Kumm, (Pleurotaceae) can ameliorate oxidative damage in aged rats. Male Wistar rats were divided into three groups of six each: group 1, normal young rats; group 2, normal aged untreated rats; group 3, normal aged rats treated with P. ostreatus (200 mg/kg body wt administered intraperitoneally for 21 days). On the 22nd day, rats were sacrificed by decapitation; the liver, kidneys, heart and brain were removed from each rat for the biochemical and isozyme analyses of the antioxidant enzymes glucose 6-phosphate dehydrogenase (G6PDH), ascorbate peroxidase (Apx) and xanthine dehydrogenase (XDH). An elevated activity of XDH was observed in the liver (G2:13.72 ± 4.1 versus G1: 7.57 ± 1.15; p ostreatus to aged rats resulted in decreased XDH and increased G6PDH and Apx activities in liver, kidneys, heart and brain. Interestingly, analyses of isozyme pattern of these enzymes are support the results obtained from the spectrophotometric determinations. These results suggest that an extract of P. ostreatus can protect the age-related oxidative damage in major organs of Wistar rats by enhancing the antioxidant enzymes G6PDH and Apx and by reducing XDH.

  8. A novel major gene on chromosome 6H for resistance of barley against the barley yellow dwarf virus

    NARCIS (Netherlands)

    Niks, R.E.; Habekuss, A.; Bekele, B.; Ordon, F.

    2004-01-01

    In a mapping population derived from the Ethiopian barley line L94 x Vada, natural infection by barley yellow dwarf virus (BYDV) occurred. While line L94 hardly showed symptoms, Vada was severely affected. The 103 recombinant inbred lines segregated bimodally. The major gene responsible for this res

  9. Accelerated evolution in the protein-coding regions is universal in crotalinae snake venom gland phospholipase A2 isozyme genes.

    Science.gov (United States)

    Nakashima, K; Nobuhisa, I; Deshimaru, M; Nakai, M; Ogawa, T; Shimohigashi, Y; Fukumaki, Y; Hattori, M; Sakaki, Y; Hattori, S

    1995-06-06

    The nucleotide sequences of four genes encoding Trimeresurus gramineus (green habu snake, crotalinae) venom gland phospholipase A2 (PLA2; phosphatidylcholine 2-acylhydrolase, EC 3.1.1.4) isozymes were compared internally and externally with those of six genes encoding Trimeresurus flavoviridis (habu snake, crotalinae) venom gland PLA2 isozymes. The numbers of nucleotide substitutions per site (KN) for the noncoding regions including introns were one-third to one-eighth of the numbers of nucleotide substitutions per synonymous site (KS) for the protein-coding regions of exons, indicating that the noncoding regions are much more conserved than the protein-coding regions. The KN values for the introns were found to be nearly equivalent to those of introns of T. gramineus and T. flavoviridis TATA box-binding protein genes, which are assumed to be a general (nonvenomous) gene. Thus, it is evident that the introns of venom gland PLA2 isozyme genes have evolved at a similar rate to those of nonvenomous genes. The numbers of nucleotide substitutions per nonsynonymous site (KA) were close to or larger than the KS values for the protein-coding regions in venom gland PLA2 isozyme genes. All of the data combined reveal that Darwinian-type accelerated evolution has universally occurred only in the protein-coding regions of crotalinae snake venom PLA2 isozyme genes.

  10. REGULATING FUNCTION OF COENZYMIZATION AND DECOENZYMIZATION OF THE LACTATE DEHYDROGENASE ISOZYMES IN THE MOUSE TISSUES DURING HYPOXIA

    Institute of Scientific and Technical Information of China (English)

    袁明秀; 蒋晖; 邓爱萍; 周晴中

    2003-01-01

    Objective. To study the characteristics of changes of LDH enzyme patterns of mice under slight hypoxia.Methods. Mice treated with artificial hypoxia, various tissues were made for the test of LDH enzy-matic activity by the specific staining technique. LDH (1-5) relative percentage enzymatic activity(RPEA) were measured with CS-910 dual-wavelength thin layer chromatography scanner.Results. The RPEA of LDH isozymes of various tissues after slight hypoxia shifted to the isozymesLDH1 and LDH2, whose principal subunits are H subunits, and the RPEA of LDH1(H4), LDH2(H3M)increased, while RPEA of LDH5(M4) in vahous tissues decreased prominently except the cardiac muscle,and that of LDH4(HM3) decreased as well. After polyacrylamide gel electrophoresis (PAGE) of the hypox-ia treated cardiac muscle specimen was made, activity subbands originated regularly in the isozyme pat-terns of LDH, with the regularity of LDH1 (0 subband), LDH2 (0-1 subbands), LDH3 (0-2 subbands),LDH4 (1-3 subbands), LDHs (2-4 subbands). After adding appropriate amount of NAD+ to the hypoxiatreated cardiac muscle specimen, PAGE showed the subbands of four isozymes (LDH2-LDH5) reduced oreven totally disappeared in the isozyme patterns.Conclusions. The negative feedback regulation of coenzymization and decoenzymization of LDHisozymes is one of the mouse stress responses to slight hypoxia.

  11. 孔雀草POD同工酶及主要观赏性状研究%Study on Peroxidase Isozyme and Main Ornamental Characteristics of Tagetes patula Breeds

    Institute of Scientific and Technical Information of China (English)

    梁顺祥; 郭洋楠; 唐道城; 张恩

    2009-01-01

    采用垂直平板聚丙烯酰胺凝胶电泳(PAGE)方法对6个孔雀草品种的过氧化物酶(POD)同工酶进行了研究,并观测了其主要观赏性状,初步探讨了两者间的关系.结果表明:6个孔雀草品种间的POD同工酶酶谱存在不同程度的差异,同一品种不同器官的POD同工酶间存在明显的差异;分析了6个品种的主要观赏性状,雪域的综合表现最好;主要观赏性状与POD同工酶酶谱间未表现出明显的规律性的关系.

  12. Amino acid sequence of Coprinus macrorhizus peroxidase and cDNA sequence encoding Coprinus cinereus peroxidase. A new family of fungal peroxidases.

    Science.gov (United States)

    Baunsgaard, L; Dalbøge, H; Houen, G; Rasmussen, E M; Welinder, K G

    1993-04-01

    Sequence analysis and cDNA cloning of Coprinus peroxidase (CIP) were undertaken to expand the understanding of the relationships of structure, function and molecular genetics of the secretory heme peroxidases from fungi and plants. Amino acid sequencing of Coprinus macrorhizus peroxidase, and cDNA sequencing of Coprinus cinereus peroxidase showed that the mature proteins are identical in amino acid sequence, 343 residues in size and preceded by a 20-residue signal peptide. Their likely identity to peroxidase from Arthromyces ramosus is discussed. CIP has an 8-residue, glycine-rich N-terminal extension blocked with a pyroglutamate residue which is absent in other fungal peroxidases. The presence of pyroglutamate, formed by cyclization of glutamine, and the finding of a minor fraction of a variant form lacking the N-terminal residue, indicate that signal peptidase cleavage is followed by further enzymic processing. CIP is 40-45% identical in amino-acid sequence to 11 lignin peroxidases from four fungal species, and 42-43% identical to the two known Mn-peroxidases. Like these white-rot fungal peroxidases, CIP has an additional segment of approximately 40 residues at the C-terminus which is absent in plant peroxidases. Although CIP is much more similar to horseradish peroxidase (HRP C) in substrate specificity, specific activity and pH optimum than to white-rot fungal peroxidases, the sequences of CIP and HRP C showed only 18% identity. Hence, CIP qualifies as the first member of a new family of fungal peroxidases. The nine invariant residues present in all plant, fungal and bacterial heme peroxidases are also found in CIP. The present data support the hypothesis that only one chromosomal CIP gene exists. In contrast, a large number of secretory plant and fungal peroxidases are expressed from several peroxidase gene clusters. Analyses of three batches of CIP protein and of 49 CIP clones revealed the existence of only two highly similar alleles indicating less

  13. Genomic Prediction of Barley Hybrid Performance

    Directory of Open Access Journals (Sweden)

    Norman Philipp

    2016-07-01

    Full Text Available Hybrid breeding in barley ( L. offers great opportunities to accelerate the rate of genetic improvement and to boost yield stability. A crucial requirement consists of the efficient selection of superior hybrid combinations. We used comprehensive phenotypic and genomic data from a commercial breeding program with the goal of examining the potential to predict the hybrid performances. The phenotypic data were comprised of replicated grain yield trials for 385 two-way and 408 three-way hybrids evaluated in up to 47 environments. The parental lines were genotyped using a 3k single nucleotide polymorphism (SNP array based on an Illumina Infinium assay. We implemented ridge regression best linear unbiased prediction modeling for additive and dominance effects and evaluated the prediction ability using five-fold cross validations. The prediction ability of hybrid performances based on general combining ability (GCA effects was moderate, amounting to 0.56 and 0.48 for two- and three-way hybrids, respectively. The potential of GCA-based hybrid prediction requires that both parental components have been evaluated in a hybrid background. This is not necessary for genomic prediction for which we also observed moderate cross-validated prediction abilities of 0.51 and 0.58 for two- and three-way hybrids, respectively. This exemplifies the potential of genomic prediction in hybrid barley. Interestingly, prediction ability using the two-way hybrids as training population and the three-way hybrids as test population or vice versa was low, presumably, because of the different genetic makeup of the parental source populations. Consequently, further research is needed to optimize genomic prediction approaches combining different source populations in barley.

  14. Purification and characterization of soluble (cytosolic) and bound (cell wall) isoforms of invertases in barley (Hordeum vulgare) elongating stem tissue

    Science.gov (United States)

    Karuppiah, N.; Vadlamudi, B.; Kaufman, P. B.

    1989-01-01

    Three different isoforms of invertases have been detected in the developing internodes of barley (Hordeum vulgare). Based on substrate specificities, the isoforms have been identified to be invertases (beta-fructosidases EC 3.2.1.26). The soluble (cytosolic) invertase isoform can be purified to apparent homogeneity by diethylaminoethyl cellulose, Concanavalin-A Sepharose, organo-mercurial Sepharose, and Sephacryl S-300 chromatography. A bound (cell wall) invertase isoform can be released by 1 molar salt and purified further by the same procedures as above except omitting the organo-mercurial Sepharose affinity chromatography step. A third isoform of invertase, which is apparently tightly associated with the cell wall, cannot be isolated yet. The soluble and bound invertase isoforms were purified by factors of 60- and 7-fold, respectively. The native enzymes have an apparent molecular weight of 120 kilodaltons as estimated by gel filtration. They have been identified to be dimers under denaturing and nondenaturing conditions. The soluble enzyme has a pH optimum of 5.5, Km of 12 millimolar, and a Vmax of 80 micromole per minute per milligram of protein compared with cell wall isozyme which has a pH optimum of 4.5, Km of millimolar, and a Vmax of 9 micromole per minute per milligram of protein.

  15. Structure of soybean seed coat peroxidase: a plant peroxidase with unusual stability and haem-apoprotein interactions

    DEFF Research Database (Denmark)

    Henriksen, A; Mirza, O; Indiani, C

    2001-01-01

    Soybean seed coat peroxidase (SBP) is a peroxidase with extraordinary stability and catalytic properties. It belongs to the family of class III plant peroxidases that can oxidize a wide variety of organic and inorganic substrates using hydrogen peroxide. Because the plant enzyme is a heterogeneous...

  16. Replication of DNA during barley endosperm development

    DEFF Research Database (Denmark)

    Giese, H.

    1992-01-01

    The incorporation of [6-H-3]-thymidine into DNA of developing barley end sperm was examined by autoradiography of cross sections of seeds and DNA analysis. The majority of nuclear divisions took place in the very young endosperm, but as late as 25 days after anthesis there was evidence for DNA...... replication. The DNA content of the endosperm increases during development and in response to nitrogen application in parallel to the storage protein synthesis profile. The hordein genes were hypersensitive to DNase I treatment throughout development....

  17. Dynamic Allocation of Sugars in Barley

    Science.gov (United States)

    Cumberbatch, L. C.; Crowell, A. S.; Fallin, B. A.; Howell, C. R.; Reid, C. D.; Weisenberger, A. G.; Lee, S. J.; McKisson, J. E.

    2014-03-01

    Allocation of carbon and nitrogen is a key factor for plant productivity. Measurements are carried out by tracing 11C-tagged sugars using positron emission tomography and coincidence counting. We study the mechanisms of carbon allocation and transport from carbohydrate sources (leaves) to sinks (stem, shoot, roots) under various environmental conditions such as soil nutrient levels and atmospheric CO2 concentration. The data are analyzed using a transfer function analysis technique to model transport and allocation in barley plants. The experimental technique will be described and preliminary results presented. This work was supported in part by USDOE Grant No. DE-FG02-97-ER41033 and DE-SC0005057.

  18. Transglycosylation by barley α-amylase 1

    DEFF Research Database (Denmark)

    Mótyán, János A.; Fazekas, Erika; Mori, Haruhide

    2011-01-01

    The transglycosylation activity of barley α-amylase 1 (AMY1) and active site AMY1 subsite mutant enzymes was investigated. We report here the transferase ability of the V47A, V47F, V47D and S48Y single mutants and V47K/S48G and V47G/S48D double mutant AMY1 enzymes in which the replaced amino acid...... DP 2, DP 3 and DP 5 were successfully applied to detect activity of Bacillus stearothermophilus maltogenic α-amylase, human salivary α-amylase and Bacillus licheniformis α-amylase, respectively in a fast and simple fluorometric assay....

  19. Feruloylated Arabinoxylans Are Oxidatively Cross-Linked by Extracellular Maize Peroxidase but Not by Horseradish Peroxidase

    Institute of Scientific and Technical Information of China (English)

    Sally J. Burr; Stephen C. Fry

    2009-01-01

    Covalent cross-linking of soluble extraceUular arabinoxylans in living maize cultures, which models the cross-linking of wall-bound arabinoxylans, is due to oxidation of feruloyl esters to oligoferuloyl esters and ethers. The oxidizing system responsible could be H_2O_2/peroxidase, O_2/laccase, or reactive oxygen species acting non-enzymically. To distinguish these possibilities, we studied arabinoxylan cross-linking in vivo and in vitro. In living cultures, exogenous, soluble, extra-cellular, feruloylated [pentosyl-~3H]arabinoxylans underwent cross-linking, beginning abruptly 8 d after sub-culture. Cross-linking was suppressed by iodide, an H_2O_2 scavenger, indicating dependence on endogenous H2O2. However, exogenous H_2O_2 did not cause precocious cross-linking, despite the constant presence of endogenous peroxidases, suggesting that younger cultures contained natural cross-linking inhibitors. Dialysed culture-filtrates cross-linked [~3H]arabinoxylans in vitro only if H_20_2 was also added, indicating a peroxiclase requirement. This cross-linking was highly ionic-strength-dependent. The peroxidases responsible were heat-labile, although relatively heat-stable peroxidases (assayed on o-dianisidine) were also present, Surprisingly, added horseradish peroxidase, even after heat-denaturation, blocked the arabinoxylan-cross-linking action of maize peroxidases, suggesting that the horseradish protein was a competing substrate for [~3H]arabino-xylan coupling. In conclusion, we show for the first time that cross-linking of extracellular arabinoxylan in living maize cultures is an action of apoplastic peroxidases, some of whose unusual properties we report.

  20. [Response of isozyme and stress indexes of Coptis chinensis to UV-B radiation].

    Science.gov (United States)

    Wen, Quan; Zhang, Nan; Cao, Rui-Xia; Zhou, Xin-Yu; Tang, Juan; Wu, Neng-Biao

    2012-03-01

    To study the physiological mechanism of anti-stress of Coptic chinensis and provide theoretical basis for its cultivation and promoting its quality. Different degrees of the range of time and intensity of UV-B radiation were set in the experiment. Used the technique of polyacrylamide gelatin vertical board electrophoresis (PAGE) to analyse the isozyme and related stress index. The isoenzymic bands of SOD1 (Rf = 0.125), SOD2 (Rf = 0.312), CAT1 (Rf = 0.428), POD3 (Rf = 0.290), POD4 (Rf = 0.636) were induced by UV-B radiation after 3 hours, with the increase of the time of UV-B radiation, those isoenzymic bands was going to vanish or became unclear. Moreover, isoenzymic bands of CAT1 (Rf = 0.428), POD3 (Rf = 0.290) disappeared in advance under heavy intensity of UV-B radiation. Furthermore, the contents of MDA, soluble sugar, proline were higher dramatically than those of control group under UV-B radiation. However, excluding the increases of proline in UL group, the content of MDA, soluble sugar, proline of other groups commenced to decrease slowly and isoenzymic bands of soluble protein increase after 7 hours of UV-B radiation. The increase of the expression of antioxidase isozyme, accumulation of soluble sugar, soluble protein and other antioxidase matter is induced by the short-time UV-B radiation, which can protect Coptis chinensis from being harmed by UV-B radiation. However, regulation system of Coptis chinensis are broken, metabolism is disordered, the bands of antioxidase isozyme vanish or weaken, the bands of soluble protein are increased and widened, these phenomenon is caused by 7 hours of UV-B radiation.

  1. Reducing isozyme competition increases target fatty acid accumulation in seed triacylglycerols of transgenic Arabidopsis.

    Science.gov (United States)

    van Erp, Harrie; Shockey, Jay; Zhang, Meng; Adhikari, Neil D; Browse, John

    2015-05-01

    One goal of green chemistry is the production of industrially useful fatty acids (FAs) in crop plants. We focus on hydroxy fatty acids (HFAs) and conjugated polyenoic FAs (α-eleostearic acids [ESAs]) using Arabidopsis (Arabidopsis thaliana) as a model. These FAs are found naturally in seed oils of castor (Ricinus communis) and tung tree (Vernicia fordii), respectively, and used for the production of lubricants, nylon, and paints. Transgenic oils typically contain less target FA than that produced in the source species. We hypothesized that competition between endogenous and transgenic isozymes for substrates limits accumulation of unique FAs in Arabidopsis seeds. This hypothesis was tested by introducing a mutation in Arabidopsis diacylglycerol acyltransferase1 (AtDGAT1) in a line expressing castor FA hydroxylase and acyl-Coenzyme A:RcDGAT2 in its seeds. This led to a 17% increase in the proportion of HFA in seed oil. Expression of castor phospholipid:diacylglycerol acyltransferase 1A in this line increased the proportion of HFA by an additional 12%. To determine if our observations are more widely applicable, we investigated if isozyme competition influenced production of ESA. Expression of tung tree FA conjugase/desaturase in Arabidopsis produced approximately 7.5% ESA in seed lipids. Coexpression of VfDGAT2 increased ESA levels to approximately 11%. Overexpression of VfDGAT2 combined with suppression of AtDGAT1 increased ESA accumulation to 14% to 15%. Our results indicate that isozyme competition is a limiting factor in the engineering of unusual FAs in heterologous plant systems and that reduction of competition through mutation and RNA suppression may be a useful component of seed metabolic engineering strategies.

  2. BIOCHEMICAL GENETIC STUDIES ON CUTTLEFISH SEPIELLA MAINDRONI (CEPHALOPODA: SEPIIDAE)- ACTIVE LOCI SCREENING OF ISOZYME

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Screening of 46 putative enzyme-coding loci and 4 different kinds of tissues of Sepiella maindroni de Rochebrone, 1884 for enzymatic activities using starch gel electrophoretic technique proved that the 21 enzymes such as AAT, AK, ALP, AP, CK, DIA, ES, FBP, G3PDH, GPI, GRS,IDH, LDH, MDH, MEP, MPI, NP, PGDH, PGM, SOD and XO* , were active to Sepiella maindroni after being stained. The tissue exhibiting stable and clear bands was also determined. Among tissues tested, mantle muscle tissue was the best for electrophoretic survey of isozymes. Buccal bulb muscle, eye and liver were fairly good for some special enzymes, such as DIA, ES, MPI, NP, etc.

  3. Ostensible enzyme promiscuity: alkene cleavage by peroxidases.

    Science.gov (United States)

    Mutti, Francesco G; Lara, Miguel; Kroutil, Markus; Kroutil, Wolfgang

    2010-12-17

    Enzyme promiscuity is generally accepted as the ability of an enzyme to catalyse alternate chemical reactions besides the 'natural' one. In this paper peroxidases were shown to catalyse the cleavage of a C=C double bond adjacent to an aromatic moiety for selected substrates at the expense of molecular oxygen at an acidic pH. It was clearly shown that the reaction occurs due to the presence of the enzyme; furthermore, the reactivity was clearly linked to the hemin moiety of the peroxidase. Comparison of the transformations catalysed by peroxidase and by hemin chloride revealed that these two reactions proceed equally fast; additional experiments confirmed that the peptide backbone was not obligatory for the reaction and only a single functional group of the enzyme was required, namely in this case the prosthetic group (hemin). Consequently, we propose to define such a promiscuous activity as 'ostensible enzyme promiscuity'. Thus, we call an activity that is catalysed by an enzyme 'ostensible enzyme promiscuity' if the reactivity can be tracked back to a single catalytic site, which on its own can already perform the reaction equally well in the absence of the peptide backbone.

  4. Redox thermodynamics of lactoperoxidase and eosinophil peroxidase.

    Science.gov (United States)

    Battistuzzi, Gianantonio; Bellei, Marzia; Vlasits, Jutta; Banerjee, Srijib; Furtmüller, Paul G; Sola, Marco; Obinger, Christian

    2010-02-01

    Eosinophil peroxidase (EPO) and lactoperoxidase (LPO) are important constituents of the innate immune system of mammals. These heme enzymes belong to the peroxidase-cyclooxygenase superfamily and catalyze the oxidation of thiocyanate, bromide and nitrite to hypothiocyanate, hypobromous acid and nitrogen dioxide that are toxic for invading pathogens. In order to gain a better understanding of the observed differences in substrate specificity and oxidation capacity in relation to heme and protein structure, a comprehensive spectro-electrochemical investigation was performed. The reduction potential (E degrees ') of the Fe(III)/Fe(II) couple of EPO and LPO was determined to be -126mV and -176mV, respectively (25 degrees C, pH 7.0). Variable temperature experiments show that EPO and LPO feature different reduction thermodynamics. In particular, reduction of ferric EPO is enthalpically and entropically disfavored, whereas in LPO the entropic term, which selectively stabilizes the oxidized form, prevails on the enthalpic term that favors reduction of Fe(III). The data are discussed with respect to the architecture of the heme cavity and the substrate channel. Comparison with published data for myeloperoxidase demonstrates the effect of heme to protein linkages and heme distortion on the redox chemistry of mammalian peroxidases and in consequence on the enzymatic properties of these physiologically important oxidoreductases.

  5. Characterization of volatile aroma compounds in different brewing barley cultivars.

    Science.gov (United States)

    Dong, Liang; Hou, Yingmin; Li, Feng; Piao, Yongzhe; Zhang, Xiao; Zhang, Xiaoyu; Li, Cheng; Zhao, Changxin

    2015-03-30

    Beer is a popular alcoholic malt beverage resulting from fermentation of the aqueous extract of malted barley with hops. The aroma of brewing barley impacts the flavor of beer indirectly, because some flavor compounds or their precursors in beer come from the barley. The objectives of this research were to study volatile profiles and to characterize odor-active compounds of brewing barley in order to determine the variability of the aroma composition among different brewing barley cultivars. Forty-one volatiles comprising aldehydes, ketones, alcohols, organic acids, aromatic compounds and furans were identified using solid phase microextraction combined with gas chromatography/mass spectrometry, among which aldehydes, alcohols and ketones were quantitatively in greatest abundance. Quantitative measurements performed by means of solvent extraction and calculation of odor activity values revealed that acetaldehyde, 2-methylpropanal, 3-methylbutanal, 2-methylbutanal, hexanal, heptanal, octanal, nonanal, 3-methyl-1-butanol, cyclopentanol, 2,3-butanedione, 2,3-pentanedione, 2-heptanone, acetic acid, ethyl acetate, 2-pentylfuran and benzeneacetaldehyde, whose concentrations exceeded their odor thresholds, could be considered as odor-active compounds of brewing barley. Principal component analysis was employed to evaluate the differences among cultivars. The results demonstrated that the volatile profile based on the concentrations of aroma compounds enabled good differentiation of most barley cultivars. © 2014 Society of Chemical Industry.

  6. Influence of Temperature on the Extractibility of Polysaccharides in Barley

    Directory of Open Access Journals (Sweden)

    Rodica Căpriţă

    2011-10-01

    Full Text Available Barley contains substantial amounts of both soluble and insoluble non-starch polysaccharides (NSP. The main watersoluble NSP in barley are highly viscous β-glucans. Monogastric animals, including humans and birds, cannotsynthesize β-glucanase, and the amount of β-glucanase derived from barley grain and bacteria in the gastrointestinaltract is insufficient to completely hydrolyze β-glucans. In the present investigation, we have studied the influence oftemperature and heating time on the extractibility of soluble polysaccharides in barley. Heating the barley samples at60°C and 80°C before extraction has the effect of lowering the soluble fraction of the polysaccharides. The dynamicviscosity values of water extracts from barley decreased up to 21.68% when heating at 60ºC for 15 minutes, and upto 25.30% when heating at 80ºC for 15 minutes, when the determinations were made immediately after extractseparation. Heating the barley samples for 15 minutes at 80°C deactivates the endogenous hydrolytic enzymes.

  7. Fungal laccase, manganese peroxidase and lignin peroxidase: gene expression and regulation.

    Science.gov (United States)

    Janusz, Grzegorz; Kucharzyk, Katarzyna H; Pawlik, Anna; Staszczak, Magdalena; Paszczynski, Andrzej J

    2013-01-10

    Extensive research efforts have been dedicated to characterizing expression of laccases and peroxidases and their regulation in numerous fungal species. Much attention has been brought to these enzymes broad substrate specificity resulting in oxidation of a variety of organic compounds which brings about possibilities of their utilization in biotechnological and environmental applications. Research attempts have resulted in increased production of both laccases and peroxidases by the aid of heterologous and homologous expression. Through analysis of promoter regions, protein expression patterns and culture conditions manipulations it was possible to compare and identify common pathways of these enzymes' production and secretion. Although laccase and peroxidase proteins have been crystallized and thoroughly analyzed, there are still a lot of questions remaining about their evolutionary origin and the physiological functions. This review describes the present understanding of promoter sequences and correlation between the observed regulatory effects on laccase, manganese peroxidase and lignin peroxidase genes transcript levels and the presence of specific response elements. Copyright © 2012 Elsevier Inc. All rights reserved.

  8. Methane emissions from feedlot cattle fed barley or corn diets.

    Science.gov (United States)

    Beauchemin, K A; McGinn, S M

    2005-03-01

    Methane emitted from the livestock sector contributes to greenhouse gas emissions worldwide. Understanding the variability in enteric methane production related to diet is essential to decreasing uncertainty in greenhouse gas emission inventories and to identifying viable greenhouse gas reduction strategies. Our study focused on measuring methane in growing beef cattle fed corn- or barley-based diets typical of those fed to cattle in North American feedlots. The experiment was designed as a randomized complete block (group) design with two treatments, barley and corn. Angus heifer calves (initial BW = 328 kg) were allocated to two groups (eight per group), with four cattle in each group fed a corn or barley diet. The experiment was conducted over a 42-d backgrounding phase, a 35-d transition phase and a 32-d finishing phase. Backgrounding diets consisted of 70% barley silage or corn silage and 30% concentrate containing steam-rolled barley or dry-rolled corn (DM basis). Finishing diets consisted of 9% barley silage and 91% concentrate containing barley or corn (DM basis). All diets contained monensin (33 mg/kg of DM). Cattle were placed into four large environmental chambers (two heifers per chamber) during each phase to measure enteric methane production for 3 d. During the backgrounding phase, DMI was greater by cattle fed corn than for those fed barley (10.2 vs. 7.6 kg/d, P cattle were in the chambers; thus, methane emissions (g/d) reported may underestimate those of the feedlot industry. Methane emissions per kilogram of DMI and as a percentage of GE intake were not affected by grain source during the backgrounding phase (24.6 g/kg of DMI; 7.42% of GE), but were less (P methane emissions of cattle fed high-forage backgrounding diets and barley-based finishing diets. Mitigating methane losses from cattle will have long-term environmental benefits by decreasing agriculture's contribution to greenhouse gas emissions.

  9. STUDIES ON SYNBIOTIC BARLEY GRAIN EXTRACT AGAINST SOME HUMAN PATHOGENS

    Directory of Open Access Journals (Sweden)

    T. Sheela

    2012-01-01

    Full Text Available This study evaluated that effect of prebiotic food containing oligosaccharide to enhance the growth and activity of probiotic strains. Barley grains probioticated using different strains of probiotics are Lactobacillus kefiranofaciens, Candida kefir,and saccharomyces boluradii. To select a suitable prebiotics like inulin for the development of Synbiotic barley and tested for antibacterial activity against diarrhoea causing pathogen such as Esherichia coli, Staphylococcus aureus, Salmonella paratyphi A, Shigella dysenteriae, Vibrio cholerae. Analysis of identified compound from synbiotic barley grain using GC-MS.

  10. Comparison of beer quality attributes between beers brewed with 100% barley malt and 100% barley raw material.

    Science.gov (United States)

    Steiner, Elisabeth; Auer, Andrea; Becker, Thomas; Gastl, Martina

    2012-03-15

    Brewing with 100% barley using the Ondea® Pro exogenous brewing enzyme product was compared to brewing with 100% barley. The use of barley, rather than malt, in the brewing process and the consequences for selected beer quality attributes (foam formation, colloidal stability and filterability, sensory differences, protein content and composition) was considered. The quality attributes of barley, malt, kettle-full-wort, cold wort, unfiltered beer and filtered beer were assessed. A particular focus was given to monitoring changes in the barley protein composition during the brewing process and how the exogenous OndeaPro® enzymes influenced wort protein composition. All analyses were based on standard brewing methods described in ASBC, EBC or MEBAK. To monitor the protein changes two-dimensional polyacrylamide gel electrophoresis was used. It was shown that by brewing beer with 100% barley and an appropriate addition of exogenous Ondea® Pro enzymes it was possible to efficiently brew beer of a satisfactory quality. The production of beers brewed with 100% barley resulted in good process efficiency (lautering and filtration) and to a final product whose sensory quality was described as light, with little body and mouthfeel, very good foam stability and similar organoleptic qualities compared to conventional malt beer. In spite of the sensory evaluation differences could still be seen in protein content and composition. Copyright © 2011 Society of Chemical Industry.

  11. Identification of the Ndh (NAD(P)H-plastoquinone-oxidoreductase) complex in etioplast membranes of barley: changes during photomorphogenesis of chloroplasts.

    Science.gov (United States)

    Guéra, A; de Nova, P G; Sabater, B

    2000-01-01

    In the last few years the presence in thylakoid membranes of chloroplasts of a NAD(P)H-plastoquinone oxidoreductase complex (Ndh complex) homologous to mitochondrial complex I has been well established. Herein, we report the identification of the Ndh complex in barley etioplast membranes. Two plastid DNA-encoded polypeptides of the Ndh complex (NDH-A and NDH-F) were relatively more abundant in etioplast membranes than in thylakoids from greening chloroplasts. Conversion of etioplast into chloroplast, after light exposure of barley seedlings grown in the dark, was accompanied by a decrease in the NADH dehydrogenase activity associated to plastid membranes. Using native-PAGE and immunolabelling techniques we have determined that a NADH specific dehydrogenase activity associated with plastid membranes, which was more active in etioplasts than in greening chloroplasts, contained the NDH-A and NDH-F polypeptides. These results complemented by those obtained through blue-native-PAGE indicated that NDH-A and NDH-F polypeptides are part of a 580 kDa NADH dependent dehydrogenase complex present in etioplast membranes. This finding proves that accumulation of the Ndh complex is independent of light. The decrease in the relative levels and specific activity of this complex during the transition from etioplast to chloroplasts was accompanied by a parallel decrease in the specific activity of peroxidase associated to plastid membranes. Based on the mentioned observations it is proposed that an electron transport chain from NADH to H2O2 could be active in barley etioplasts.

  12. Isozyme and allozyme markers distinguishing two morphologically similar, medically important Mastomys species (Rodentia: Muridae

    Directory of Open Access Journals (Sweden)

    Van der Bank Herman FH

    2001-09-01

    Full Text Available Abstract Background Two common southern African mice species, Mastomys coucha and M. natalensis, are widely distributed throughout the subregion and overlap in many areas. They also share a high degree of morphological similarity, making them impossible to distinguish in the field at present. These multimammate mice are documented carriers of serious disease vectors causing Lassa fever, plague and encephalomyocarditis, which coupled to their cohabitation with humans in many areas, could pose a significant health risk. A preliminary study reported the presence of isozyme markers at three loci (GPI-2, PT-2, -3 in one population each of M. coucha and M. natalensis. Two additional populations (from the Vaal Dam and Richards Bay were sampled to determine the reliability of these markers, and to seek additional genetic markers. Results Fifteen proteins or enzymes provided interpretable results at a total of 39 loci. Additional fixed allele differences between the species were detected at AAT-1, ADH, EST-1, PGD-1, Hb-1 and -2. Average heterozygosities for M. coucha and M. natalensis were calculated as 0.018 and 0.032 respectively, with a mean genetic distance between the species of 0.26. Conclusions The confirmation of the isozyme and the detection of the additional allozyme markers are important contributions to the identification of these two medical and agricultural pest species.

  13. Inhibitory properties of nerve-specific human glutamate dehydrogenase isozyme by chloroquine.

    Science.gov (United States)

    Choi, Myung-Min; Kim, Eun-A; Choi, Soo Young; Kim, Tae Ue; Cho, Sung-Woo; Yang, Seung-Ju

    2007-11-30

    Human glutamate dehydrogenase exists in hGDH1 (housekeeping isozyme) and in hGDH2 (nerve-specific isozyme), which differ markedly in their allosteric regulation. In the nervous system, GDH is enriched in astrocytes and is important for recycling glutamate, a major excitatory neurotransmitter during neurotransmission. Chloroquine has been known to be a potent inhibitor of house-keeping GDH1 in permeabilized liver and kidney-cortex of rabbit. However, the effects of chloroquine on nerve-specific GDH2 have not been reported yet. In the present study, we have investigated the effects of chloroquine on hGDH2 at various conditions and showed that chloroquine could inhibit the activity of hGDH2 at dose-dependent manner. Studies of the chloroquine inhibition on enzyme activity revealed that hGDH2 was relatively less sensitive to chloroquine inhibition than house-keeping hGDH1. Incubation of hGDH2 was uncompetitive with respect of NADH and non-competitive with respect of 2-oxoglutarate. The inhibitory effect of chloroquine on hGDH2 was abolished, although in part, by the presence of ADP and L-leucine, whereas GTP did not change the sensitivity to chloroquine inhibition. Our results show a possibility that chloroquine may be used in regulating GDH activity and subsequently glutamate concentration in the central nervous system.

  14. Transmembrane carbonic anhydrase isozymes IX and XII in the female mouse reproductive organs

    Directory of Open Access Journals (Sweden)

    Tomas Eija

    2004-10-01

    Full Text Available Abstract Background Carbonic anhydrase (CA classically catalyses the reversible hydration of dissolved CO2 to form bicarbonate ions and protons. The twelve active CA isozymes are thought to regulate a variety of cellular functions including several processes in the reproductive systems. Methods The present study was designed to investigate the expression of transmembrane CAs, CA IX and XII, in the mouse uterus, ovary and placenta. The expression of CA IX and XII was examined by immunoperoxidase staining method and western blotting. CA II and XIII served as positive controls since they are known to be present in the mouse reproductive tract. Results The data of our study indicated that CA XII is expressed in the mouse endometrium. Only very faint signal was observed in the corpus luteum of the ovary and the placenta remained mainly negative. CA IX showed weak reaction in the endometrial epithelium, while it was completely absent in the ovary and placenta. Conclusion The conservation of CA XII expression in both mouse and human endometrium suggests a role for this isozyme in reproductive physiology.

  15. Isozymic variations in specific and nonspecific esterase and its thermostability in silkworm, Bombyx mori L.

    Science.gov (United States)

    Patnaik, Bharat Bhusan; Biswas, Tapati Datta; Nayak, Sandeepta Kumar; Saha, A K; Majumdar, M K

    2012-09-01

    Esterase isozymic variations were documented in the haemolymph of developed multivoltine and bivoltine silkworm breeds during unfavorable seed crop seasons of May - September using á- and â- napthylacetate separately to identify specific and nonspecific esterase having thermotolerant potentiality. Variations existed in the isozyme pattern with three bands (Est-2, 3 and 4) in pure Nistari race and other developed multivoltine and bivoltine breeds. Est-2 and Est-3 were non-specific esterases as they were observed when both á- and â-napthylacetate was used as substrates separately. Est-4 band was observed only with á-napthylacetate as substrate and was therefore confirmed to be specific á-esterase band in the haemolymph of silkworm, Bombyx mori L. Zymograms showed that the non-specific esterase band (Est-3) with R1 of 0.43 and specific á-esterase band (Est-4) with R(f) of 0.32 predominately withstood a temperature of 70 +/- 2 degrees C for a duration of 10 min and were confirmed as thermostable esterases in haemolymph of silkworm, Bombyx mori L. This also categorized the presence of thermostable esterases in developed multivoltine and bivoltine breeds of silkworm, even though the qualitative activity was more in the former than the latter. The qualitative presence of thermostable esterases and their activity could be adopted as an indicative biochemical marker in relation to thermotolerance in silkworm.

  16. Expression of rapeseed microsomal lysophosphatidic acid acyltransferase isozymes enhances seed oil content in Arabidopsis.

    Science.gov (United States)

    Maisonneuve, Sylvie; Bessoule, Jean-Jacques; Lessire, René; Delseny, Michel; Roscoe, Thomas J

    2010-02-01

    In higher plants, lysophosphatidic acid acyltransferase (LPAAT), located in the cytoplasmic endomembrane compartment, plays an essential role in the synthesis of phosphatidic acid, a key intermediate in the biosynthesis of membrane phospholipids in all tissues and storage lipids in developing seeds. In order to assess the contribution of LPAATs to the synthesis of storage lipids, we have characterized two microsomal LPAAT isozymes, the products of homoeologous genes that are expressed in rapeseed (Brassica napus). DNA sequence homologies, complementation of a bacterial LPAAT-deficient mutant, and enzymatic properties confirmed that each of two cDNAs isolated from a Brassica napus immature embryo library encoded a functional LPAAT possessing the properties of a eukaryotic pathway enzyme. Analyses in planta revealed differences in the expression of the two genes, one of which was detected in all rapeseed tissues and during silique and seed development, whereas the expression of the second gene was restricted predominantly to siliques and developing seeds. Expression of each rapeseed LPAAT isozyme in Arabidopsis (Arabidopsis thaliana) resulted in the production of seeds characterized by a greater lipid content and seed mass. These results support the hypothesis that increasing the expression of glycerolipid acyltransferases in seeds leads to a greater flux of intermediates through the Kennedy pathway and results in enhanced triacylglycerol accumulation.

  17. Novel dehydroepiandrosterone benzimidazolyl derivatives as 5α-reductase isozymes inhibitors.

    Science.gov (United States)

    Arellano, Yazmín; Bratoeff, Eugene; Segura, Tania; Mendoza, Maria Eugenia; Sánchez-Márquez, Araceli; Medina, Yesica; Heuze, Yvonne; Soriano, Juan; Cabeza, Marisa

    2016-12-01

    5α-R isozymes (types 1 and 2) play an important role in prostate gland development because they are responsible for intraprostatic dihydrotestosterone (DHT) levels when the physiological serum testosterone (T) concentration is low. In this study, we synthesized seven novel dehydroepiandrosterone derivatives with benzimidazol moiety at C-17, and determined their effect on the activity of 5α-reductase types 1 and 2. The derivatives with an aliphatic ester at C-3 of the dehydroepiandrosterone scaffold induced specific inhibition of 5α-R1 activity, whereas those with a cycloaliphatic ester (cyclopropyl, cyclobutyl, or cyclopentyl ring) or an alcohol group at C-3 inhibited the activity of both isozymes. Derivatives with a cyclohexyl or cycloheptyl ester at C-3 showed no inhibitory activity. In pharmacological experiments, derivatives with esters having an alcohol or the aliphatic group or one of the three smaller cycloaliphatic rings at C-3 decreased the diameter of male hamster flank organs, with the cyclobutyl and cyclopentyl esters exhibiting higher effect. With exception of the cyclobutyl and cyclopentyl esters, these compounds reduced the weight of the prostate and seminal vesicles.

  18. Isozyme Analysis of Jin Silver Carp ( Hypophthalmichthys molitrix Var Jin)

    Institute of Scientific and Technical Information of China (English)

    Qiang YANG; Jun HAO; Di BAO; Aijun LIANG; Wankun JIN; Chongwen LI; Xinghua ZHANG; Shi DONG

    2012-01-01

    Abstract [Objective] The aim was to carry out isozyme analysis of jin silver carp (Hypophthalmichthys mofitrix Var Jing). [Method] The isozyme of AAT, EST, cc-GPD, GPI, IDH, LDH, MDH, ME, PGM and PROT of muscles and liver in two populations of the silver carp (Hypophthalmichthys molitrix): Jin silver carp (a breed through se- lective breeding) and artificially propagated population bought from Jingzhou city, Hubei Province were examined by horizontal starch gel electrophoresis. [Result] Eigh- teen loci were observed in two populations. Two loci of GPI and PGM in Jing sil- ver carp population and the locus of GPI in Jingzhou population were polymorphic. The proportions of polymorphic loci (maximum allele frequency-〈0.99) of Jing silver carp and Jingzhou populations were 11.11% and 5.56% respectively, expected het- erozygosity were 0.015 0 and 0.001 1 respectively. The Nei's genetic distances were 0.000 59 between two populations. The result of chi-square test of the GPI gene in two populations showed that their genetic structure has very significant dif- ference. [Conclusion] This study provided a theoretical basis for large-scale extension of Jing silver carp.

  19. Studies on isozymic variation among the South Indian species of Sphaerostephanos

    Institute of Scientific and Technical Information of China (English)

    Irudayaraj Varaprasadham; Johnson Marimuthu

    2011-01-01

    Objective: To explore the identity and phylogenetic relationships among the three medicinally important species of Sphaerostephanos from South India using isozymic profile. Methods: The young fronds were homogenized with 3.5 mL of ice-cold homogenizing buffer in a pre-chilled pestle and mortar. The supernatant was subjected to electrophoresis as described by Anbalagan poly acrylamide gel electrophoresis. Staining solutions for isoperoxidase was prepared as per Smila method for the detection of isoenzymes. Results: A total of six different bands in five different positions with different molecular weight/Rf values and four active zones have been observed in the isoperoxidase enzyme system of Sphaerostephanos. Only one band with MW/Rf 0.399 is common to two different species i.e. Sphaerostephanos arbuscula (S. arbuscula) andSphaerostephanos unitus (S. unitus). Among the remaining four bands, two bands (Rf. 0.23, 0.47) are present in Sphaerostephanos subtruncatus (S. subtruncatus) and one distinct band has been observed individually in S. arbuscula (Rf. 0.507) and S. unitus (Rf. 0.56). Conclusions: The present preliminary molecular study through isozymic analysis shows the identity of all the three species and the present results confirm distinctness of these three species based on macro-micromorphology, phytochemistry and cytology.

  20. Role of PKC isozymes in low-power light-stimulated proliferation of cultured skin cells

    Science.gov (United States)

    Grossman, Nili; Kleitman, Vered; Meller, Julia; Kaufmann, Roland; Akgun, Nermin; Ruck, Angelika; Livneh, Etta; Lubart, Rachel

    2000-11-01

    Exposure of cultured skin cells to low power visible light leads to a transiently stimulated proliferation. Facilitation of this response requires the presence of active PKC, elevation of intracellular calcium, and involves reactive oxygen species. In the present study, the role of PKC(alpha) and PCK(eta) was examined using paired murine fibroblasts, differing in the level of these isozymes expression. The ability of the cells to respond to low power UVA light or HeNe laser by stimulated proliferation was correlated with an active state or overexpression of PKC(alpha) , but not PKC(eta) . A parallel response was obtained in cells that were loaded with A1PcS4 before photosensitization. Whenever this latter treatment caused a light-stimulated inhibition, it was accompanied by the intracellular calcium and photosensitizer dynamics typical of the effect of PDT on rate epithelial cells. Accordingly, added antioxidants that suppressed light-stimulated proliferation also suppressed this light-stimulated inhibition. The model systems employed in this study are the first to demonstrate the specific effect of PKC isozymes on light-stimulated proliferation, in relation to oxidative stress, and indicate their dual role in light-tissue interaction.

  1. A barley SKP1-like protein controls abundance of the susceptibility factor RACB and influences the interaction of barley with the barley powdery mildew fungus.

    Science.gov (United States)

    Reiner, Tina; Hoefle, Caroline; Hückelhoven, Ralph

    2016-02-01

    In an increasing number of plant-microbe interactions, it has become evident that the abundance of immunity-related proteins is controlled by the ubiquitin-26S proteasome system. In the interaction of barley with the biotrophic barley powdery mildew fungus Blumeria graminis f.sp. hordei (Bgh), the RAC/ROP [RAT SARCOMA-related C3 botulinum toxin substrate/RAT SARCOMA HOMOLOGUE (RHO) of plants] guanosine triphosphatase (GTPase) HvRACB supports the fungus in a compatible interaction. By contrast, barley HvRBK1, a ROP-binding receptor-like cytoplasmic kinase that interacts with and can be activated by constitutively activated HvRACB, limits fungal infection success. We have identified a barley type II S-phase kinase 1-associated (SKP1)-like protein (HvSKP1-like) as a molecular interactor of HvRBK1. SKP1 proteins are subunits of the SKP1-cullin 1-F-box (SCF)-E3 ubiquitin ligase complex that acts in the specific recognition and ubiquitination of protein substrates for subsequent proteasomal degradation. Transient induced gene silencing of either HvSKP1-like or HvRBK1 increased protein abundance of constitutively activated HvRACB in barley epidermal cells, whereas abundance of dominant negative RACB only weakly increased. In addition, silencing of HvSKP1-like enhanced the susceptibility of barley to haustorium establishment by Bgh. In summary, our results suggest that HvSKP1-like, together with HvRBK1, controls the abundance of HvRACB and, at the same time, modulates the outcome of the barley-Bgh interaction. A possible feedback mechanism from RAC/ROP-activated HvRBK1 on the susceptibility factor HvRACB is discussed.

  2. Effect of phytase supplementation to barley-canola meal and barley-soybean meal diets on phosphorus and calcium balance in growing pigs

    NARCIS (Netherlands)

    Sauer, W.C.; Cervantes, M.; He, J.M.M.; Schulze, H.

    2003-01-01

    Two metabolism experiments were carried out, to determine the effect of microbial phytase addition to barley-canola meal and barley-soybean meal diets on P and Ca balance in growing. pigs; In experiment 1, six barrows (29.6kg: initial LW) were fed a barley-canola meal diet, without or. with phytase

  3. DyP, a unique dye-decolorizing peroxidase, represents a novel heme peroxidase family: ASP171 replaces the distal histidine of classical peroxidases.

    Science.gov (United States)

    Sugano, Yasushi; Muramatsu, Riichi; Ichiyanagi, Atsushi; Sato, Takao; Shoda, Makoto

    2007-12-14

    DyP, a unique dye-decolorizing enzyme from the fungus Thanatephorus cucumeris Dec 1, has been classified as a peroxidase but lacks homology to almost all other known plant peroxidases. The primary structure of DyP shows moderate sequence homology to only two known proteins: the peroxide-dependent phenol oxidase, TAP, and the hypothetical peroxidase, cpop21. Here, we show the first crystal structure of DyP and reveal that this protein has a unique tertiary structure with a distal heme region that differs from that of most other peroxidases. DyP lacks an important histidine residue known to assist in the formation of a Fe4+ oxoferryl center and a porphyrin-based cation radical intermediate (compound I) during the action of ubiquitous peroxidases. Instead, our tertiary structural and spectrophotometric analyses of DyP suggest that an aspartic acid and an arginine are involved in the formation of compound I. Sequence analysis reveals that the important aspartic acid and arginine mentioned above and histidine of the heme ligand are conserved among DyP, TAP, and cpop21, and structural and phylogenetic analyses confirmed that these three enzymes do not belong to any other families of peroxidase. These findings, which strongly suggest that DyP is a representative heme peroxidase from a novel family, should facilitate the identification of additional new family members and accelerate the classification of this novel peroxidase family.

  4. Differences in phytase activity and phytic acid content between cultivated and Tibetan annual wild barleys.

    Science.gov (United States)

    Dai, Fei; Qiu, Long; Xu, Yang; Cai, Shengguan; Qiu, Boyin; Zhang, Guoping

    2010-11-24

    The Qinghai-Tibetan Plateau in China is considered to be one of the original centers of cultivated barley. At present, little is known about the phytase activity (Phy) or phytic acid content (PA) in grains of Tibetan annual wild barley. Phy and PA were determined in grains of 135 wild and 72 cultivated barleys. Phy ranged from 171.3 to 1299.2 U kg(-1) and from 219.9 to 998.2 U kg(-1) for wild and cultivated barleys, respectively. PA and protein contents were much higher in wild barley than in cultivated barley. Tibetan annual wild barley showed a larger genetic diversity in phytase activity and phytic acid and protein contents and is of value for barley breeding. There is no significant correlation between phytase activity and phytic acid or protein content in barley grains, indicating that endogenous phytase activity had little effect on the accumulation of phytic acid.

  5. Localisation of morphological traits on the genetic map of potato using RFLP and isozyme markers = [Localisatie van morfologische eigenschappen op de genetische kaart van de aardappel door middel van RFLP en isozym merkers

    NARCIS (Netherlands)

    Eck, van H.J.

    1995-01-01


    The thesis describes the construction of a genetic linkage map of the potato genome, comprising molecular, isozyme and morphological markers. The linkage map is based on the offspring from non-inbred parents. The computer program JOINM

  6. Amylolytic strains of Lactobacillus plantarum isolated from barley

    African Journals Online (AJOL)

    aghomotsegin

    2015-01-28

    Jan 28, 2015 ... Key words: Lactobacillus plantarum, starch hydrolysis, barley, malting. ... especially in environments rich in glucose or disac- charides such as sucrose ..... numbers produce less lactic acid, which in turn is less stringent on ...

  7. Implementation of biochemical screening to improve baking quality of barley

    DEFF Research Database (Denmark)

    Vincze, Éva; Dionisio, Giuseppe; Aaslo, Per;

    2011-01-01

    Barley (Hordeum vulgare) has the potential to offer considerable human nutritional benefits, especially as supplement to wheat-based breads. Under current commercial baking conditions it is not possible to introduce more that 20% barley flour to the wheat bread without negative impact...... proteins. Changing the storage protein composition can lessen this problem. Our working hypothesis was that exploiting the substantial genetic variation within the gene pool for storage proteins could enable improving the baking qualities of barley flour. We characterised forty-nine barley cultivars...... for variations in storage protein and AA composition. These cultivars were selected based on their higher protein contents (11.8–17.6%). The results obtained indicated that substantial variation not only in the distribution of the hordein polypeptides but also in the relative proportions of the storage proteins...

  8. Evaluation of fermented whole crop wheat and barley feeding on ...

    African Journals Online (AJOL)

    이창희

    2017-07-11

    Jul 11, 2017 ... After maize, wheat and barley are produced in large quantities and account ... Through fermentation, beneficial bacteria are increased and harmful .... LDL cholesterol, triglyceride, cortisol, and blood urea nitrogen (BUN).

  9. Immobilization of Peroxidase onto Magnetite Modified Polyaniline

    Directory of Open Access Journals (Sweden)

    Eduardo Fernandes Barbosa

    2012-01-01

    Full Text Available The present study describes the immobilization of horseradish peroxidase (HRP on magnetite-modified polyaniline (PANImG activated with glutaraldehyde. After the optimization of the methodology, the immobilization of HRP on PANImG produced the same yield (25% obtained for PANIG with an efficiency of 100% (active protein. The optimum pH for immobilization was displaced by the effect of the partition of protons produced in the microenvironment by the magnetite. The tests of repeated use have shown that PANImG-HRP can be used for 13 cycles with maintenance of 50% of the initial activity.

  10. Hydrothermal liquefaction of barley straw to bio-crude oil

    DEFF Research Database (Denmark)

    Zhu, Zhe; Rosendahl, Lasse; Toor, Saqib;

    2015-01-01

    Hydrothermal liquefaction (HTL) of barley straw with K2CO3 at different temperatures (280–400 C) was conducted and compared to optimize its process conditions; the aqueous phase as a co-product from this process was recycled to explore the feasibility of implementing wastewater reuse for bio...... a detailed insight into the HTL behavior of barley straw, and offers potential opportunities and benefits for bio-crude oil production through the reuse of aqueous phase....

  11. Androgenesis in anther culture of Lithuanian spring barley cultivars

    OpenAIRE

    Asakavičiūtė, Rita; Pašakinskienė, Izolda

    2006-01-01

    The method of anther culture was used for the production of doubled haploids in Lithuanian spring barley cultivars. Two methods, (i) regeneration from callus (Szarjeko’s method) and (ii) direct regeneration from embryoids (Caredda’s method) were applied to determine the androgenic potential according to the green regenerant yield and other morphogenetic factors. Green double haploid regenerants were obtained in four Lithuanian spring barley cultivars (‘Aura’, ‘Aidas’, ‘Alsa’ and ‘Auksiniai’) ...

  12. Weed suppression ability of spring barley varieties

    DEFF Research Database (Denmark)

    Christensen, Svend

    1995-01-01

    Three years of experiments with spring barley showed significant differences in weed suppression ability among varieties. Weed dry matter in the most suppressive variety, Ida, was 48% lower than the mean weed dry matter of all varieties, whereas it was 31% higher in the least suppressive variety......, Grit. Ranking varietal responses to weed competition in terms of grain yield loss corresponded well to ranking weed dry matter produced in crop weed mixtures. There was no correspondence between the varietal grain yields in pure stands and their competitiveness, suggesting that breeding to optimize...... interception model was developed to describe the light interception profiles of the varieties. A study of the estimated parameters showed significant correlation between weed dry matter, rate of canopy height development and the light interception profile. However, when estimates were standardized to eliminate...

  13. The spontaneous chlorophyll mutation frequency in barley

    DEFF Research Database (Denmark)

    Jørgensen, Jørgen Helms; Jensen, Hans Peter

    1986-01-01

    A total of 1866 barley plants were progeny tested in the greenhouse. Twenty-five plants segregated for newly arisen, spontaneous chlorophyll mutant genes. Among the total of 470,129 seedlings screened there were 79 mutants (1.7 .+-. 0.6 .times. 10-4). The data are added to data from three similar...... materials and the resulting estimate of the chlorophyll mutant frequency is 1.6 .times. 10-4 in about 1.43 million seedlings. The estimate of the chlorophyll mutation rate per generation is close to 67.3 .times. 10-4 per diploid genome or in the order of 6 .times. 10-7 per locus and haploid genome....

  14. The Barley Chromosome 5 Linkage Map

    DEFF Research Database (Denmark)

    Jensen, J.; Jørgensen, Jørgen Helms

    1975-01-01

    The literature is surveyed for data on recombination between loci on chromosome 5 of barley; 13 loci fall into the category “mapped” loci, more than 20 into the category “associated” loci and nine into the category “loci once suggested to be on chromosome 5”. A procedure was developed...... for estimating a linkage map; it involves (1) transformation by the Kosambi mapping function of the available recombination percentages to additive map distances, (2) calculations of a set of map distances from the transformed recombination percentages by a maximum likelihood method in which all the available...... data are utilized jointly, and (3) omission of inconsistent data and determination of the most likely order of the loci. This procedure was applied to the 42 recombination percentages available for the 13 “mapped” loci. Due to inconsistencies 14 of the recombination percentages and, therefore, two...

  15. Development of endosperm transfer cells in barley

    Directory of Open Access Journals (Sweden)

    Johannes eThiel

    2014-03-01

    Full Text Available Endosperm transfer cells (ETCs are positioned at the intersection of maternal and filial tissues in seeds of cereals and represent a bottleneck for apoplasmic transport of assimilates into the endosperm. Endosperm cellularization starts at the maternal-filial boundary and generates the highly specialized ETCs. During differentiation barley ETCs develop characteristic flange-like wall ingrowths to facilitate effective nutrient transfer. A comprehensive morphological analysis depicted distinct developmental time points in establishment of transfer cell morphology and revealed intracellular changes possibly associated with cell wall metabolism. Embedded inside the grain, ETCs are barely accessible by manual preparation. To get tissue-specific information about ETC specification and differentiation, laser microdissection(LM-based methods were used for transcript and metabolite profiling. Transcriptome analysis of ETCs at different developmental stages by microarrays indicated activated gene expression programs related to control of cell proliferation and cell shape, cell wall and carbohydrate metabolism reflecting the morphological changes during early ETC development. Transporter genes reveal distinct expression patterns suggesting a switch from active to passive modes of nutrient uptake with the onset of grain filling. Tissue-specific RNA-seq of the differentiating ETC region from the syncytial stage until functionality in nutrient transfer identified a high number of novel transcripts putatively involved in ETC differentiation. An essential role for two-component signaling (TCS pathways in transfer cell development of barley emerged from this analysis. Correlative data provide evidence for ABA and ethylene influences on ETC differentiation and hint at a crosstalk between hormone signal transduction and TCS phosphorelays. Collectively, the data expose a comprehensive view on ETC development, associated pathways and identified candidate genes for

  16. Global identification and analysis of isozyme-specific possible substrates crosslinked by transglutaminases using substrate peptides in mouse liver fibrosis

    Science.gov (United States)

    Tatsukawa, Hideki; Tani, Yuji; Otsu, Risa; Nakagawa, Haruka; Hitomi, Kiyotaka

    2017-01-01

    The transglutaminase (TG) family comprises eight isozymes that form the isopeptide bonds between glutamine and lysine residues and contribute to the fibrotic diseases via crosslinking-mediated stabilization of ECM and the activation of TGF-β in several tissues. However, despite a growing body of evidence implicating TG2 as a key enzyme in fibrosis, the causative role of TG2 and the involvement of the other isozymes have not yet been fully elucidated. Therefore, here we clarified the distributions of TG isozymes and their in situ activities and identified the isozyme-specific possible substrates for both TG1 and TG2 using their substrate peptides in mouse fibrotic liver. We found that TG1 activity was markedly enhanced intracellularly over a widespread area, whereas TG2 activity increased in the extracellular space. In total, 43 and 42 possible substrates were identified for TG1 and TG2, respectively, as involved in chromatin organization and cellular component morphogenesis. These included keratin 18, a biomarker for hepatic injury, which was accumulated in the fibrotic liver and showed the partly similar distribution with TG1 activity. These findings suggest that TG1 activity may be involved in the functional modification of intracellular proteins, whereas TG2 activity contributes to the stabilization of extracellular proteins during liver fibrosis. PMID:28327670

  17. 3D structure prediction of lignolytic enzymes lignin peroxidase and manganese peroxidase based on homology modelling

    Directory of Open Access Journals (Sweden)

    SWAPNIL K. KALE

    2016-04-01

    Full Text Available Lignolytic enzymes have great biotechnological value in biopulping, biobleaching, and bioremediation. Manganese peroxidase (EC 1:11:1:13 and lignin peroxidase (EC 1:11:1:14 are extracellular and hem-containing peroxidases that catalyze H2O2-dependent oxidation of lignin. Because of their ability to catalyse oxidation of a wide range of organic compounds and even some inorganic compounds, they got tremendous industrial importance. In this study, 3D structure of lignin and manganese peroxidase has been predicted on the basis of homology modeling using Swiss PDB workspace. The physicochemical properties like molecular weight, isoelectric point, Grand average of hydropathy, instability and aliphatic index of the target enzymes were performed using Protparam. The predicted secondary structure of MnP has 18 helices and 6 strands, while LiP has 20 helices and 4 strands. Generated 3D structure was visualized in Pymol. The generated model for MnP and LiP has Z-score Qmean of 0.01 and -0.71, respectively. The predicted models were validated through Ramachandran Plot, which indicated that 96.1 and 95.5% of the residues are in most favored regions for MnP and LiP respectively. The quality of predicted models were assessed and confirmed by VERIFY 3D, PROCHECK and ERRAT. The modeled structure of MnP and LiP were submitted to the Protein Model Database.

  18. The effect of added enzymes on process potentials derived from different qualities of barley

    DEFF Research Database (Denmark)

    Shetty, Radhakrishna; Zhuang, Shiwen; Hansen, Preben Bøje;

    Barley sorting is an important step for picking up grain of desired quality. Whilst brewing with 100% sorted barley (picked high quality) has become realistic with the addition of exogenous enzymes, the effect of added enzymes on process potentials derived from un-sorted barley (mixed) and sorted...... filterability, the Ondea® Pro treatment resulted in significantly lower turbidity and smaller particle size compared to Cellic® CTec2; however, this effect was observed in sorted and un-sorted barley but not in sorted-out barley. Consequently the un-sorted barley demonstrated great potential in brewing process...

  19. Long-term reconstitution of dry barley increased phosphorus digestibility in pigs

    DEFF Research Database (Denmark)

    Ton Nu, Mai Anh; Blaabjerg, Karoline; Poulsen, Hanne Damgaard

    of reconstitution compared to dry stored barley on phosphorus (P) digestibility in pigs. Materials and Methods: Dry barley (13% moisture; phytate P, 1.7 g/kg DM) was rolled and stored directly or reconstituted with water to produce rolled barley with 35% moisture that was stored in air-tight conditions. After 49......: Reconstituted barley had higher soluble P (2.56 g/kg DM) and lower phytate P (0.93 g/ kg DM) compared with dry barley (0.78 and 1.7 g/kg DM, respectively). Pigs fed the reconstituted barley diet showed increased P absorption (52%) and decreased P excretion in feces (21%) (P

  20. A PRELIMINARY STUDY OF CERVICOVAGINAL PEROXIDASES AS INDICATORS FOR OVULATION

    Institute of Scientific and Technical Information of China (English)

    XIANGHong-Fa; HANZi-Yan; LIANGZang-Guang; XIESu-Xiang

    1989-01-01

    There were many studies using cervicovaginal peroxidases to predict ovulation. Some resuits suggested that cervieovaginal peroxidases are reliable indicators for ovulation; but others did not. The present study was designed to determine whether the change patterns of ccrvicovaginal guaiacul peroxidase activity in fertile period of Chinese women can also be served as a basis for development of a technique to predict ovulation time in natural family planning.

  1. Constructing the barley model for genetic transformation in Triticeae

    Institute of Scientific and Technical Information of China (English)

    LÜ Bo; WU Jia-jie; FU Dao-lin

    2015-01-01

    Barley (Hordeum vulgare L.) is one of the oldest domesticated crops, showing dramatic adaptation to various climate and environmental conditions. As a major cereal crop, barley ranks the 4th after wheat, maize and rice in terms of planting area and production al over the world. Due to its diploid nature, the cultivated barley is considered as an ideal model to study the polyploid wheat and other Triticeae species. Here, we reviewed the development, optimization, and application of transgenic approaches in barley. The most efifcient and robust genetic transformation has been built on the Agrobacterium-mediated transfer in conjunction with the immature embryo-based regeneration. We then discussed future considerations of using more practical technologies in barley transformation, such as the T-DNA/transposon tagging and the genome editing. As a cereal crop amenable to genetic transformation, barley wil serve as the most valuable carrier for global functional genomics in Triticeae and is becoming the most practical model for generating value-added products.

  2. Glycaemic response to barley porridge varying in dietary fibre content.

    Science.gov (United States)

    Thondre, Pariyarath S; Wang, Ke; Rosenthal, Andrew J; Henry, Christiani J K

    2012-03-01

    The interest in barley as a food is increasing worldwide because of its high dietary fibre (DF) content and low glycaemic index (GI). DF in cereals may prove beneficial in improving blood glucose response in the long term. However, a dose-dependent effect of insoluble fibre on reducing postprandial blood glucose levels is yet to be proven. The objective of the present study was to determine the glycaemic response to two barley porridges prepared from whole barley grains varying in fibre content. In two separate non-blind randomised crossover trials, ten human subjects consumed barley porridge with 16 g/100 g and 10 g/100 g fibre content provided in different serving sizes (equivalent to 25 and 50 g available carbohydrate). The glycaemic response to both barley porridges was significantly lower than the reference glucose (P porridges. We concluded that irrespective of the difference in total fibre content or serving size of barley porridges, their GI values did not differ significantly.

  3. Initial-rate kinetics of human NMN-adenylyltransferases: substrate and metal ion specificity, inhibition by products and multisubstrate analogues, and isozyme contributions to NAD+ biosynthesis.

    Science.gov (United States)

    Sorci, Leonardo; Cimadamore, Flavio; Scotti, Stefania; Petrelli, Riccardo; Cappellacci, Loredana; Franchetti, Palmarisa; Orsomando, Giuseppe; Magni, Giulio

    2007-04-24

    Initial-rate and product inhibition studies revealed distinctive ordered ternary complex kinetic mechanisms, substrate specificities, and metal ion preferences for the three isozymes of human nicotinamide mononucleotide adenylyl-transferase (NMNAT, EC 2.7.7.1). ATP binds before NMN with nuclear isozyme NMNAT1 and Golgi apparatus NMNAT2, but the opposite order is observed with the mitochondrial isozyme NMNAT3. Only the latter utilizes ITP efficiently in place of ATP, and while NMNH conversion to NADH by NMNAT1 and NMNAT3 occurs at similar rates, conversion by NMNAT2 is much slower. These isozymes can also be discriminated by their action on tiazofurin monophosphate (TrMP), a metabolite of the antineoplastic prodrug tiazofurin. Our finding that TrMP is only a substrate with NMNAT1 and NMNAT3 reveals for the first time an organelle selectivity in the metabolism of this important drug. In search of additional ways to discriminate these isozymes, we synthesized and tested the P1-(nicotinamide/nicotinate-riboside-5')-Pn-(adenosine-5') dinucleotides Np3AD, Np4AD, and Nap4AD. In addition to being highly effective inhibitors, these multisubstrate geometric inhibitors gave inhibition patterns that are consistent with the aforementioned isozyme differences in substrate binding order. Distinctive differences in their substrate specificity and metal ion selectivity also permitted us to quantify individual isozyme contributions to NAD+ formation in human cell extracts.

  4. Roles of apoplastic peroxidases in plant response to wounding.

    Science.gov (United States)

    Minibayeva, Farida; Beckett, Richard Peter; Kranner, Ilse

    2015-04-01

    Apoplastic class III peroxidases (EC 1.11.1.7) play key roles in the response of plants to pathogen infection and abiotic stresses, including wounding. Wounding is a common stress for plants that can be caused by insect or animal grazing or trampling, or result from agricultural practices. Typically, mechanical damage to a plant immediately induces a rapid release and activation of apoplastic peroxidases, and an oxidative burst of reactive oxygen species (ROS), followed by the upregulation of peroxidase genes. We discuss how plants control the expression of peroxidases genes upon wounding, and also the sparse information on peroxidase-mediated signal transduction pathways. Evidence reviewed here suggests that in many plants production of the ROS that comprise the initial oxidative burst results from a complex interplay of peroxidases with other apoplastic enzymes. Later responses following wounding include various forms of tissue healing, for example through peroxidase-dependent suberinization, or cell death. Limited data suggest that ROS-mediated death signalling during the wound response may involve the peroxidase network, together with other redox molecules. In conclusion, the ability of peroxidases to both generate and scavenge ROS plays a key role in the involvement of these enigmatic enzymes in plant stress tolerance. Copyright © 2014 Elsevier Ltd. All rights reserved.

  5. Nematode assemblages in the rhizosphere of spring barley (Hordeum vulgare L.) depended on fertilisation and plant growth phase

    DEFF Research Database (Denmark)

    Madsen, Mette Vestergård

    2004-01-01

    rhizosphere; nitrogen and phosphorus fertilisation; nematode assemblages; plant parasites; barley......rhizosphere; nitrogen and phosphorus fertilisation; nematode assemblages; plant parasites; barley...

  6. Competition and Facilitation in Hairy Vetch-Barley Intercrops

    Directory of Open Access Journals (Sweden)

    Giacomo Tosti

    2010-09-01

    Full Text Available Intercrops between legumes and non-legumes are widely used for fodder production and as cover crops, but little quantitative data are available on competition between species in the mixture. The objective of the present study was to assess the interaction between hairy vetch (Vicia villosa Roth. and barley (Hordeum vulgare L. grown as pure crops or intercrops with different proportions of seed rates at sowing. A 4-year field study was conducted using hairy vetch and barley as pure stands at full sowing density and as intercrops at different proportions of their own full seed rate according to the replacement principle. Interaction between species was evaluated on the basis of Land Equivalent Ratio (LER, Relative Neighbour Effect (RNE and Aggressivity (A calculated on biomass and nitrogen (N accumulation. The N accumulation of the mixed crops increased linearly with the legume proportion in the mixture. The mixtures were more efficient than the pure crops in terms of N use (LER > 1. Partial LER values indicated that the barley component benefited from the presence of the legume, while the hairy vetch partial LER decreased with increasing barley proportion in the mixture. The competitive response in terms of biomass accumulation was high for both species when their density in the mixture was high. Concerning N accumulation, barley benefited from an asymmetric interspecific facilitation while the vetch behaviour was similar to that observed for biomass accumulation. Barley dominance progressively increased reaching a maximum just before the last sampling date. At the last sampling date the competitive ability of hairy vetch showed a considerable increase in all mixtures (A ≈ 0. These findings indicate that the use of mixtures between hairy vetch and barley allows an increase in the use efficiency of N resource with respect to pure crops. Barley is the dominant component of the mixture and the hairy vetch is able to cope with the cereal

  7. Physiological and Ecological Studies in Environmental Adaptation of Plants. : III. Altitudinal Variation in Some Characters of Cytochrome Oxidase Isozymes in Polygonum cuspidatum Sieb. et Zucc.

    OpenAIRE

    1987-01-01

    The number and the activity of cytochrome oxidase isozymes in Polygonum cuspidatum Sieb. et Zucc. were investigated on the mature plants growing in a few different altitudes and on these seedlings grown under various levels of a constant temperature. The number of the isozymes in the mature plants was 3 at low altitude and 2 at high altitude, but that in the plants transplanted from high to low altitude was 4. The number and the activity of the isozymes in the seedlings derived from the plant...

  8. Genetic Variation of Isozyme Polyphenol Oxidase (PPO Profiles in Different Varieties of Capsicum annuum L.

    Directory of Open Access Journals (Sweden)

    Owk ANIEL KUMAR

    2013-12-01

    Full Text Available The genus Capsicum commonly known as chilli pepper is a major spice crop and is of cosmopolitan in distribution. Native polyacrylamide gel electrophoresis (Native PAGE was used to study the polyphenol oxidase (PPO isozyme variation in 21 varieties of Capsicum annuum L. A maximum of 4 PPO bands were scored in five varieties i.e., Ca14, Ca15, Ca16, Ca19 & Ca20, while the minimum (2 bands was observed in four varieties (Ca3, Ca10, Ca13 & Ca17. 15 pair wise combinations showed highest average per cent similarity (100% and the UPGMA dendrogram represented low genetic diversity. The present study revealed that considerable intraspecific differences were found in the varieties. Thus the results obtained could be used in fingerprinting the genotypes.

  9. Isozymes in Larrea divaricata and Larrea tridentata (Zygophyllaceae): a study of two amphitropical vicariants and autopolyploidy.

    Science.gov (United States)

    Cortes, M C; Hunziker, J H

    1997-10-01

    Electrophoretic variants for seven isozyme systems - probably encoded by 18 structural gene loci - in diploid populations of Larrea divaricata and diploid and tetraploid populations of its North American vicariant derivative L. tridentata were assayed by polyacrilamide and starch gel electrophoresis. High molecular similarity of diploid and tetraploid cytotypes of L. tridentata supports the hypothesis of interracial autopolyploidy. The absence of fixed heterozygosity and additive profiles indicates a low level of divergence between the parental diploids and the tetraploids. The phenogram based on the I coefficient showed the similarities between the populations of diploid L. divaricata and also between the diploid populations of L. tridentata. Both groups of diploid populations were more distantly connected to tetraploid L. tridentata.

  10. Metabolism of cellulose by Phanerochaete chrysosporium in continuously agitated culture is associated with enhanced production of lignin peroxidase.

    Science.gov (United States)

    Zacchi, L; Burla, G; Zuolong, D; Harvey, P J

    2000-03-10

    Production of the extracellular heme protein lignin peroxidase (LiP) by Phanerochaete chrysosporium is currently associated with a number of requirements, namely exposure of the cultures to oxygen; limiting nutrient nitrogen or carbon and static or semi-static culture conditions. To obtain LiP activity in continuously agitated liquid culture requires the inclusion of a surfactant. However, using cellulose as the carbon source, we obtained high titres (0.2-0.4 U ml(-1)) of LiP in submerged liquid cultures under conditions of continuous agitation, without substrate limitation or the need to add oxygen or surfactant. Comparison of the morphological and physiological traits of hyphae maintained on either cellulose or free glucose supports observations that the synthesis of extracellular polysaccharide in the cultures grown on glucose, restricts oxygen diffusion into the hyphae, which is necessary for LiP induction. They also suggest that isozymes of LiP synthesised under these conditions may be triggered in response to oxidant stress.

  11. Regional and accelerated molecular evolution in group I snake venom gland phospholipase A2 isozymes.

    Science.gov (United States)

    Chuman, Y; Nobuhisa, I; Ogawa, T; Deshimaru, M; Chijiwa, T; Tan, N H; Fukumaki, Y; Shimohigashi, Y; Ducancel, F; Boulain, J C; Ménez, A; Ohno, M

    2000-03-01

    In accordance with detection of a few phospholipase A2 (PLA2) isozyme genes by Southern blot analysis, only two cDNAs, named NnkPLA-I , and NnkPLA-II, encoding group I PLA2s, NnkPLA-I and NnkPLA-II, respectively, were isolated from the venom gland cDNA library of Elapinae Naja naja kaouthia of Malaysia. NnkPLA-I and NnkPLA-II showed four amino acid substitutions, all of which were brought about by single nucleotide substitution. No existence of clones encoding CM-II and CM-III, PLA2 isozymes which had been isolated from the venom of N. naja kaouthia of Thailand, in Malaysian N. naja kaouthia venom gland cDNA library was verified by dot blot hybridization analysis with particular probes. NnkPLA-I and NnkPLA-II differed from CM-II and CM-III with four and two amino acid substitutions, respectively, suggesting that their molecular evolution is regional. The comparison of NnkPLA-I, NnkPLA-II and cDNAs encoding other group I snake venom gland PLA2s indicated that the 5'- and 3'-untranslated regions are more conserved than the mature protein-coding region and that the number of nucleotide substitutions per nonsynonymous site is almost equal to that per synonymous site in the protein-coding region, suggesting that accelerated evolution has occurred in group I venom gland PLA2s possibly to acquire new physiological functions.

  12. Identification of CYP isozymes involved in benzbromarone metabolism in human liver microsomes.

    Science.gov (United States)

    Kobayashi, Kaoru; Kajiwara, Eri; Ishikawa, Masayuki; Oka, Hidenobu; Chiba, Kan

    2012-11-01

    Benzbromarone (BBR) is metabolized to 1'-hydroxy BBR and 6-hydroxy BBR in the liver. 6-Hydroxy BBR is further metabolized to 5,6-dihydroxy BBR. The aim of this study was to identify the CYP isozymes involved in the metabolism of BBR to 1'-hydroxy BBR and 6-hydroxy BBR and in the metabolism of 6-hydroxy BBR to 5,6-dihydroxy BBR in human liver microsomes. Among 11 recombinant P450 isozymes examined, CYP3A4 showed the highest formation rate of 1'-hydroxy BBR. The formation rate of 1'-hydroxy BBR significantly correlated with testosterone 6β-hydroxylation activity in a panel of 12 human liver microsomes. The formation of 1'-hydroxy BBR was completely inhibited by ketoconazole in pooled human liver microsomes. On the other hand, the highest formation rate of 6-hydroxy BBR was found in recombinant CYP2C9. The highest correlation was observed between the formation rate of 6-hydroxy BBR and diclofenac 4'-hydroxylation activity in 12 human liver microsomes. The formation of 6-hydroxy BBR was inhibited by tienilic acid in pooled human liver microsomes. The formation of 5,6-dihydroxy BBR from 6-hydroxy BBR was catalysed by recombinant CYP2C9 and CYP1A2. The formation rate of 5,6-dihydroxy BBR was significantly correlated with diclofenac 4'-hydroxylation activity and phenacetin O-deethylation activity in 12 human liver microsomes. The formation of 5,6-dihydroxy BBR was inhibited with either tienilic acid or α-naphthoflavone in human liver microsomes. These results suggest that (i) the formation of 1'-hydroxy BBR and 6-hydroxy BBR is mainly catalysed by CYP3A4 and CYP2C9, respectively, and (ii) the formation of 5,6-dihydroxy BBR is catalysed by CYP2C9 and CYP1A2 in human liver microsomes.

  13. Efficient production of tetraploid barley (Hordeum vulgare L. by colchicine treatment of diploid barley

    Directory of Open Access Journals (Sweden)

    Ayed Sourour

    2014-03-01

    Full Text Available An experiment was conducted to induce tetraploidy in three diploid barley varieties (Martin, Rihane and Manel through different colchicines treatments. Colchicine was added for three different concentrations at three different stages of plant development i.e. on seed (0.05% for 48 hours, on pre-germinated seeds (0.1% for 2 hours and on three leaf stage (0.1% for 16 hours. Colchicine application reduced significantly germination percentage and viability of plants. Seed germination was completely inhibited in Martin, while a reduction of 20% and 30% for germination percentage compared to control was recorded in varieties Manel and Rihane, respectively at 0.1% colchicine concentration. Ploidy evaluation showed no tetraploidy in all the three tested varieties by colchicine application of 0.05% for 48 hours on seeds and 0.1% for 2 hours on pre-germinated seeds. However, tetraploid plants were produced only by treatment with 0.1% for 16 hours of seedlings. The percentages of plants were 40%, 44% and 100% for Rihane, Manel and Martin, respectively. Cytological analyses showed the increase of chromosome numbers from 2n=2x=14 to 2n=4x=28. The increase of ploidy levels caused major changes in some morphological traits. In fact, the induced tetraploids in barley was accompanied by significant (P<0.01 decrease in plant height, tiller height, leaf number and leaf length compared to diploid control plants. colchicine treatment induce successfully the production of tetraploid barley plants and could be used in breeding programs.

  14. Molecular characterization of barley 3H semi-dwarf genes.

    Directory of Open Access Journals (Sweden)

    Haobing Li

    Full Text Available The barley chromosome 3H accommodates many semi-dwarfing genes. To characterize these genes, the two-rowed semi-dwarf Chinese barley landrace 'TX9425' was crossed with the Australian barley variety 'Franklin' to generate a doubled haploid (DH population, and major QTLs controlling plant height have been identified in our previous study. The major QTL derived from 'TX9425' was targeted to investigate the allelism of the semi-dwarf gene uzu in barley. Twelve sets of near-isogenic lines and a large NILF2 fine mapping population segregating only for the dwarfing gene from 'TX9425' were developed. The semi-dwarfing gene in 'TX9425' was located within a 2.8 cM region close to the centromere on chromosome 3H by fine mapping. Molecular cloning and sequence analyses showed that the 'TX9425'-derived allele contained a single nucleotide substitution from A to G at position 2612 of the HvBRI1 gene. This was apparently the same mutation as that reported in six-rowed uzu barley. Markers co-segregating with the QTL were developed from the sequence of the HvBRI1 gene and were validated in the 'TX9425'/'Franklin' DH population. The other major dwarfing QTL derived from the Franklin variety was distally located on chromosome 3HL and co-segregated with the sdw1 diagnostic marker hv20ox2. A third dwarfing gene, expressed only in winter-sown trials, was identified and located on chromosome 3HS. The effects and interactions of these dwarfing genes under different growing conditions are discussed. These results improve our understanding of the genetic mechanisms controlling semi-dwarf stature in barley and provide diagnostic markers for the selection of semi-dwarfness in barley breeding programs.

  15. Cytochrome c as a peroxidase : tuning of heme reactivity

    NARCIS (Netherlands)

    Diederix, Rutger Ernest Michiel

    2003-01-01

    This thesis describes the peroxidase activity of the electron-transfer protein cytochrome c, and how it is controlled by the protein matrix. It is shown that unfolding cytochrome c has the effect to significantly enhance its peroxidase activity of (up to several thousand-fold). This can be achieved

  16. ATP-enhanced peroxidase-like activity of gold nanoparticles.

    Science.gov (United States)

    Shah, Juhi; Purohit, Rahul; Singh, Ragini; Karakoti, Ajay Singh; Singh, Sanjay

    2015-10-15

    Gold nanoparticles (AuNPs) are known to possess intrinsic biological peroxidase-like activity that has applications in development of numerous biosensors. The reactivity of the Au atoms at the surface of AuNPs is critical to the performance of such biosensors, yet little is known about the effect of biomolecules and ions on the peroxidase-like activity. In this work, the effect of ATP and other biologically relevant molecules and ions over peroxidase-like activity of AuNPs are described. Contrary to the expectation that nanoparticles exposed to biomolecules may lose the catalytic property, ATP and ADP addition enhanced the peroxidase-like activity of AuNPs. The catalytic activity was unaltered by the addition of free phosphate, sulphate and carbonate anions however, addition of ascorbic acid to the reaction mixture diminished the intrinsic peroxidase-like activity of AuNPs, even in the presence of ATP and ADP. In contrast to AuNPs, ATP did not synergize and improve the peroxidase activity of the natural peroxidase enzyme, horseradish peroxidase.

  17. Hydrogen peroxide-mediated inactivation of two chloroplastic peroxidases, ascorbate peroxidase and 2-cys peroxiredoxin.

    Science.gov (United States)

    Kitajima, Sakihito

    2008-01-01

    Reactive oxygen species (ROS), such as the superoxide anion and hydrogen peroxide, are generated by the photosystems because photoexcited electrons are often generated in excess of requirements for CO2 fixation and used for reducing molecular oxygen, even under normal environmental conditions. Moreover, ROS generation is increased in chloroplasts if plants are subjected to stresses, such as drought, high salinity and chilling. Chloroplast-localized isoforms of ascorbate peroxidase and possibly peroxiredoxins assume the principal role of scavenging hydrogen peroxide. However, in vitro studies revealed that both types of peroxidases are easily damaged by hydrogen peroxide and lose their catalytic activities. This is one contributing factor for cellular damage that occurs under severe oxidative stress. In this review, I describe mechanisms of hydrogen peroxide-mediated inactivation of these two enzymes and discuss a reason why they became susceptible to damage by hydrogen peroxide.

  18. The relationship between lignin peroxidase and manganese peroxidase production capacities and cultivation periods of mushrooms.

    Science.gov (United States)

    Xu, Jian Z; Zhang, Jun L; Hu, Kai H; Zhang, Wei G

    2013-05-01

    Mushrooms are able to secrete lignin peroxidase (LiP) and manganese peroxidase (MnP), and able to use the cellulose as sources of carbon. This article focuses on the relation between peroxidase-secreting capacity and cultivation period of mushrooms with non-laccase activity. Methylene blue and methyl catechol qualitative assay and spectrophotometry quantitative assay show LiP secreting unvaryingly accompanies the MnP secreting in mushroom strains. The growth rates of hyphae are detected by detecting the dry hyphal mass. We link the peroxidase activities to growth rate of mushrooms and then probe into the relationship between them. The results show that there are close relationships between LiP- and/or MnP-secretory capacities and the cultivation periods of mushrooms. The strains with high LiP and MnP activities have short cultivation periods. However, those strains have long cultivation periods because of the low levels of secreted LiP and/or MnP, even no detectable LiP and/or MnP activity. This study provides the first evidence on the imitate relation between the level of secreted LiP and MnP activities and cultivation periods of mushrooms with non-laccase activity. Our study has significantly increased the understanding of the role of LiP and MnP in the growth and development of mushrooms with non-laccase activity. © 2012 The Authors. Microbial Biotechnology © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.

  19. Toxoplasma gondii: demonstration of intrinsic peroxidase activity during lacto-peroxidase mediated radioiodination of tachyzoites

    Energy Technology Data Exchange (ETDEWEB)

    Gallois, Y.; Tricaud, A.; Foussard, F.; Hodbert, J.; Girault, A.; Mauras, G.; Dubremetz, J.F.

    1986-01-01

    Tachyzoites of Toxoplasma gondii have been radioiodinated under various conditions with or without lactoperoxidase, with glucose oxidase being used to generate hydrogen peroxide. Erythrocytes were iodinated simultaneously as a control. In our conditions, tachyzoites were more intensely labelled in the absence of lactoperoxidase. This result can be explained by the existence of an intrinsic peroxidase activity which interfere with the exogenously added enzyme during surface radioiodination.

  20. Applications and Prospective of Peroxidase Biocatalysis in the Environmental Field

    Science.gov (United States)

    Torres-Duarte, Cristina; Vazquez-Duhalt, Rafael

    Environmental protection is, doubtless, one of the most important challenges for the human kind. The huge amount of pollutants derived from industrial activities represents a threat for the environment and ecologic equilibrium. Phenols and halogenated phenols, polycyclic aromatic hydrocarbons, endocrine disruptive chemicals, pesticides, dioxins, polychlorinated biphenyls, industrial dyes, and other xenobiotics are among the most important pollutants. A large variety of these xenobiotics are substrates for peroxidases and thus susceptible to enzymatic transformation. The literature reports mainly the use of horseradish peroxidase, manganese peroxidase, lignin peroxidase, and chloroperoxidase on the transformation of these pollutants. Peroxidases are enzymes able to transform a variety of compounds following a free radical mechanism, giving oxidized or polymerized products. The peroxidase transformation of these pollutants is accompanied by a reduction in their toxicity, due to a biological activity loss, a reduction in the bioavailability or due to the removal from aqueous phase, especially when the pollutant is found in water. In addition, when the pollutants are present in soil, peroxidases catalyze a covalent binding to soil organic matter. In most of cases, oxidized products are less toxic and easily biodegradable than the parent compounds. In spite of their versatility and potential use in environmental processes, peroxidases are not applied at large scale yet. Diverse challenges, such as stability, redox potential, and the production of large amounts, should be solved in order to apply peroxidases in the pollutant transformation. In this chapter, we critically review the transformation of different xenobiotics by peroxidases, with special attention on the identified transformation products, the probable reaction mechanisms, and the toxicity reports. Finally, the design and development of an environmental biocatalyst is discussed. The design challenges are

  1. Pearling barley to alter the composition of the raw material before brewing

    NARCIS (Netherlands)

    Donkelaar, van L.H.G.; Noordman, T.R.; Boom, R.M.; Goot, van der A.J.

    2015-01-01

    Partly replacing malt with unmalted barley is a trend in brewing. The use of unmalted barley, however, leads to issues such as haze and high mash viscosity, due to its higher content of undesired components. Pearling, an abrasive method to remove the outer layers of the barley kernels has been shown

  2. Interaction between powdery mildew and barley with ¤mlo5¤ mildew resistance

    DEFF Research Database (Denmark)

    Lyngkjær, M.F.; Østergård, Hanne

    1998-01-01

    Powdery mildew infection of barley with the mlo5 barley powdery mildew resistance gene was examined, using near-isogenic barley lines, with and without mlo5 resistance, and two near-isogenic powdery mildew isolates, HL3/5 and GE3 with high (virulent) or low (avirulent) penetration efficiency...

  3. 7 CFR 457.102 - Wheat or barley winter coverage endorsement.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 6 2010-01-01 2010-01-01 false Wheat or barley winter coverage endorsement. 457.102... INSURANCE CORPORATION, DEPARTMENT OF AGRICULTURE COMMON CROP INSURANCE REGULATIONS § 457.102 Wheat or barley... Wheat or Barley Winter Coverage Endorsement (This is a continuous endorsement) 1. In return for...

  4. Genetic dissection of grain beta-glucan and amylose content in barley (Hordeum vulgare L.)

    Science.gov (United States)

    High beta glucan (BG) barleys (Hordeum vulgare L.) have major potential as food ingredients due to the well know health benefits. Quantitative trait loci (QTLs) associated with BG have been reported in hulled barley, however no QTL studies have been reported in hulless barley. In this study, QTL an...

  5. Reactions of the class II peroxidases, lignin peroxidase and Arthromyces ramosus peroxidase, with hydrogen peroxide. Catalase-like activity, compound III formation, and enzyme inactivation.

    Science.gov (United States)

    Hiner, Alexander N P; Hernández-Ruiz, Josefa; Rodríguez-López, José Neptuno; García-Cánovas, Francisco; Brisset, Nigel C; Smith, Andrew T; Arnao, Marino B; Acosta, Manuel

    2002-07-26

    The reactions of the fungal enzymes Arthromyces ramosus peroxidase (ARP) and Phanerochaete chrysosporium lignin peroxidase (LiP) with hydrogen peroxide (H(2)O(2)) have been studied. Both enzymes exhibited catalase activity with hyperbolic H(2)O(2) concentration dependence (K(m) approximately 8-10 mm, k(cat) approximately 1-3 s(-1)). The catalase and peroxidase activities of LiP were inhibited within 10 min and those of ARP in 1 h. The inactivation constants were calculated using two independent methods; LiP, k(i) approximately 19 x 10(-3) s(-1); ARP, k(i) approximately 1.6 x 10(-3) s(-1). Compound III (oxyperoxidase) was detected as the majority species after the addition of H(2)O(2) to LiP or ARP, and its formation was accompanied by loss of enzyme activity. A reaction scheme is presented which rationalizes the turnover and inactivation of LiP and ARP with H(2)O(2). A similar model is applicable to horseradish peroxidase. The scheme links catalase and compound III forming catalytic pathways and inactivation at the level of the [compound I.H(2)O(2)] complex. Inactivation does not occur from compound III. All peroxidases studied to date are sensitive to inactivation by H(2)O(2), and it is suggested that the model will be generally applicable to peroxidases of the plant, fungal, and prokaryotic superfamily.

  6. The role of root hairs in cadmium acquisition by barley

    Energy Technology Data Exchange (ETDEWEB)

    Zheng Ruilun; Li Huafen [Key Laboratory of Plant-Soil Interactions of the Ministry of Education, College of Resources and Environmental Sciences, China Agricultural University, Beijing 100094 (China); Jiang Rongfeng, E-mail: rfjiang@cau.edu.c [Key Laboratory of Plant-Soil Interactions of the Ministry of Education, College of Resources and Environmental Sciences, China Agricultural University, Beijing 100094 (China); Roemheld, Volker [Institute of Plant Nutrition, University of Hohenheim, D-70593 Stuttgart (Germany); Zhang Fusuo [Key Laboratory of Plant-Soil Interactions of the Ministry of Education, College of Resources and Environmental Sciences, China Agricultural University, Beijing 100094 (China); Zhao Fangjie [Soil Science Department, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ (United Kingdom)

    2011-02-15

    The role of root hairs in Cd acquisition from soil was investigated in three pot experiments using a root hairless mutant (bald root barley, brb) and its wild-type (WT) cultivar of barley (Hordeum vulgare). brb had significantly lower concentrations and lower total amounts of Cd in shoots than WT. The Cd uptake efficiency based on total root length was 8-45% lower in brb than in WT. The difference between brb and WT increased with increasing extractable Cd in soil under the experimental conditions used. Additions of phosphate to soil decreased Cd extractability. Both soil and foliar additions of phosphate decreased root length, and root hair formation in WT. These effects resulted in decreased Cd uptake with increasing P supply. Cd uptake in WT correlated significantly with root length, root hair length and density, and soil extractable Cd. Root hairs contribute significantly to Cd uptake by barley. - Research highlights: The Cd uptake efficiency was significantly lower in brb than in WT. Additions of phosphate to soil decreased Cd extractability and Cd uptake. Both soil and foliar additions of phosphate decreased root length, and root hair formation in WT. Root hairs contribute significantly to Cd uptake by barley. - The Cd uptake efficiency based on total root length was 8-45% lower in a barley root hairless mutant than in its wild-type, indicating an important role of root hairs in Cd acquisition.

  7. Identification of a phytase gene in barley (Hordeum vulgare L..

    Directory of Open Access Journals (Sweden)

    Fei Dai

    Full Text Available BACKGROUND: Endogenous phytase plays a crucial role in phytate degradation and is thus closely related to nutrient efficiency in barley products. The understanding of genetic information of phytase in barley can provide a useful tool for breeding new barley varieties with high phytase activity. METHODOLOGY/PRINCIPAL FINDINGS: Quantitative trait loci (QTL analysis for phytase activity was conducted using a doubled haploid population. Phytase protein was purified and identified by the LC-ESI MS/MS Shotgun method. Purple acid phosphatase (PAP gene was sequenced and the position was compared with the QTL controlling phytase activity. A major QTL for phytase activity was mapped to chromosome 5 H in barley. The gene controlling phytase activity in the region was named as mqPhy. The gene HvPAP a was mapped to the same position as mqPhy, supporting the colinearity between HvPAP a and mqPhy. CONCLUSIONS/SIGNIFICANCE: It is the first report on QTLs for phytase activity and the results showed that HvPAP a, which shares a same position with the QTL, is a major phytase gene in barley grains.

  8. Mutagenesis of barley malting quality QTLs with Ds transposons.

    Science.gov (United States)

    Singh, Surinder; Tan, Han Qi; Singh, Jaswinder

    2012-03-01

    Various functional genomic tools are being used to identify and characterize genes in plants. The Activator/Dissociation (Ac/Ds) transposon-based approach offers great potential, especially in barley, due to its limited success of genetic transformation and its large genome size. The bias of the Ac/Ds system towards genic regions and its tendency toward localized transpositions can greatly enhance the discovery and tagging of genes linked to Ds. Barley is a key ingredient in malting and brewing industry; therefore, gene discovery in relation to malting has an industrial perspective. Malting quality in barley is a complex and quantitatively inherited trait. Two major quantitative trait loci (QTLs) affecting malting quality traits have been located on chromosome 4H. In this study, Ds was reactivated from parent transposants (TNP) lines, TNP-29 and TNP-79, where Ds was mapped in the vicinity of important malting QTLs. Reactivation of Ds was carried out both by conventional breeding and in vitro approaches. A threefold increase in reactivation frequency through the in vitro approach enabled the development of a new genomic resource for the dissection of malting QTL and gene discovery in barley. Identification of unique flanking sequences, using high-efficiency thermal asymmetric interlaced PCR and inverse PCR from these populations, has further emphasized the new location of Ds in the barley genome and provided new transposon mutants especially in β-GAL1, β-amylase-like gene and ABC transporter for functional genomic studies.

  9. Products based on a high fiber barley genotype, but not on common barley or oats, lower postprandial glucose and insulin responses in healthy humans.

    Science.gov (United States)

    Liljeberg, H G; Granfeldt, Y E; Björck, I M

    1996-02-01

    Postprandial blood glucose and insulin responses to cereal products made from common barley, oats or a barley genotype containing elevated levels of beta-glucans were evaluated in nine healthy subjects. Porridges were made from commercial Swedish whole-meal barley or oat flours, and a mixed whole-meal porridge using the high fiber barley genotype and commercial Swedish common barley (50:50). Also studied were two types of flour-based bread products composed of high fiber barley and common barley in ratios of 50:50 or 80:20, respectively. The common oat and barley porridges produced postprandial glucose and insulin responses similar to the white wheat bread reference, suggesting that the naturally occurring dietary fiber in these whole-meal flours has no impact on the glucose tolerance. In contrast, all high fiber barley products induced significantly lower responses than did the reference product, with the glycemic and insulin indices ranging from 57 to 72 or 42 to 72%, respectively. It is concluded that "lente" products of high sensory quality can be prepared from a barley genotype with an elevated content of soluble dietary fiber. The glycemic index of these products compares favorably with that of products made from common cereals, suggesting their use as a potential component of diets for patients with diabetes and hyperlipidemia, and for individuals predisposed to metabolic disease.

  10. Lysine metabolism in antisense C-hordein barley grains

    DEFF Research Database (Denmark)

    Schmidt, Daiana; Rizzi, Vanessa; Gaziola, Salete A

    2015-01-01

    ) and five antisense C-hordein transgenic barley lines. Considering the amounts of soluble and protein-bound aspartate-derived amino acids together with the analysis of key enzymes of aspartate metabolic pathway, we suggest that the C-hordein suppression did not only alter the metabolism of at least one......The grain proteins of barley are deficient in lysine and threonine due to their low concentrations in the major storage protein class, the hordeins, especially in the C-hordein subgroup. Previously produced antisense C-hordein transgenic barley lines have an improved amino acid composition......, with increased lysine, methionine and threonine contents. The objective of the study was to investigate the possible changes in the regulation of key enzymes of the aspartate metabolic pathway and the contents of aspartate-derived amino acids in the nontransgenic line (Hordeum vulgare L. cv. Golden Promise...

  11. Application of proteomics to investigate barley-Fusarium graminearum interaction

    DEFF Research Database (Denmark)

    Yang, Fen

    the disease. Due to the advantages of gel-based proteomics that differentially expressed proteins involved in the interaction can be directly detected by comparing protein profiles displayed on 2-D gels, it is used as a tool for studying the barley- Fusarium graminearum interaction form three different....... The functional characterization of two proteins is undergoing. In Chapter 6, microarray data of F. graminearum during interaction with barley and wheat was analysed. The expression patterns of 11fungal genes in microarray analysis were different from qRT-PCR results in Chapter 4. Overall, our results will give...... some insights into the cellular activities during the interaction between barley and Fusarium graminearum for designing new efficient strategies for the control of FHB disease....

  12. Factors underlying restricted crossover localization in barley meiosis.

    Science.gov (United States)

    Higgins, James D; Osman, Kim; Jones, Gareth H; Franklin, F Chris H

    2014-01-01

    Meiotic recombination results in the formation of cytological structures known as chiasmata at the sites of genetic crossovers (COs). The formation of at least one chiasma/CO between homologous chromosome pairs is essential for accurate chromosome segregation at the first meiotic division as well as for generating genetic variation. Although DNA double-strand breaks, which initiate recombination, are widely distributed along the chromosomes, this is not necessarily reflected in the chiasma distribution. In many species there is a tendency for chiasmata to be distributed in favored regions along the chromosomes, whereas in others, such as barley and some other grasses, chiasma localization is extremely pronounced. Localization of chiasma to the distal regions of barley chromosomes restricts the genetic variation available to breeders. Studies reviewed herein are beginning to provide an explanation for chiasma localization in barley. Moreover, they suggest a potential route to manipulating chiasma distribution that could be of value to plant breeders.

  13. Origin of worldwide cultivated barley revealed by NAM-1 gene and grain protein content

    Directory of Open Access Journals (Sweden)

    Yonggang eWang

    2015-09-01

    Full Text Available The origin, evolution and distribution of cultivated barley provides powerful insights into the historic origin and early spread of agrarian culture. Here, population-based genetic diversity and phylogenetic analyses were performed to determine the evolution and origin of barley and how domestication and subsequent introgression have affected the genetic diversity and changes in cultivated barley on a worldwide scale. A set of worldwide cultivated and wild barleys from Asia and Tibet of China were analyzed using the sequences for NAM-1 gene and gene-associated traits-GPC (grain protein content. Our results showed Tibetan wild barley distinctly diverged from Near Eastern barley, and confirmed that Tibet is one of the origin and domestication centers for cultivated barley, and in turn supported a polyphyletic origin of domesticated barley. Comparison of haplotype composition among geographic regions revealed gene flow between Eastern and Western barley populations, suggesting that the Silk Road might have played a crucial role in the spread of genes. The GPC in the 118 cultivated and 93 wild barley accessions ranged from 6.73% to 12.35% with a mean of 9.43%. Overall, wild barley had higher averaged GPC (10.44% than cultivated barley. Two unique haplotypes (Hap2 and Hap7 caused by a base mutations (at position 544 in the coding region of the NAM-1 gene might have a significant impact on the GPC. SNPs and haplotypes of NAM-1 associated with GPC in barley could provide a useful method for screening GPC in barley germplasm. The Tibetan wild accessions with lower GPC could be useful for malt barley breeding

  14. Prognostic significance of glutathione peroxidase 2 in gastric carcinoma.

    Science.gov (United States)

    Liu, Dongzhe; Sun, Liang; Tong, Jinxue; Chen, Xiuhui; Li, Hui; Zhang, Qifan

    2017-06-01

    Increasing evidence suggests that the glutathione peroxidase 2 may actually play important roles in tumorigenesis and progression in various human cancers such as colorectal carcinomas and lung adenocarcinomas. However, the role of glutathione peroxidase 2 in gastric carcinoma remains to be determined. In this study, the expression and prognostic significance of glutathione peroxidase 2 in gastric carcinoma were investigated and the well-known prognostic factor Ki-67 labeling index was also assessed as positive control. Glutathione peroxidase 2 expression levels in the tumor tissue specimens, the matched adjacent normal tissue specimens, and the lymph node metastases of 176 patients with gastric carcinoma were evaluated by quantitative polymerase chain reaction, western blotting, and immunohistochemical staining. The associations between glutathione peroxidase 2 expression levels, as determined by immunohistochemical staining, and multiple clinicopathological characteristics were determined by Pearson's chi-square test and Spearman's correlation analysis. The relationships between glutathione peroxidase 2 expression and other clinicopathological variables and patient prognoses were analyzed further by the Kaplan-Meier method, the log-rank test, and Cox multivariate regression. The quantitative polymerase chain reaction, western blotting, and immunohistochemical staining results showed that glutathione peroxidase 2 expression levels were upregulated in both the primary tumor foci and the lymph node metastases of patients with gastric carcinoma (all p values gastric carcinoma (all p values gastric carcinoma that may be used to devise personalized therapeutic regimens and precision treatments for this disease.

  15. Purification and characterization of peroxidase from papaya (Carica papaya) fruit.

    Science.gov (United States)

    Pandey, Veda P; Singh, Swati; Singh, Rupinder; Dwivedi, Upendra N

    2012-05-01

    Ripening of papaya fruit was found to be characterized with a decrease in peroxidase activity and its transcript. This peroxidase was purified to homogeneity through successive steps of ammonium sulfate fractionation, ion exchange and molecular exclusion chromatography. The peroxidase was purified 30.22-folds with overall recovery of 44.37% and specific activity of 68.59. Purified peroxidase was found to be a heterotrimer of ~240 kDa, containing two subunits each of 85 and one of 70 kDa. Purified enzyme exhibited pH and temperature optima of 7.0 and 40 °C, respectively. K(m) values for substrates o-dianicidin, guaiacol and ascorbic acid were found to be 0.125, 0.8 and 5.2 mM, respectively. K(m) for H(2)O(2) was found to be 0.25 mM. Salicylic acid was found to activate peroxidase up to 50 μM concentration, beyond which it acted as inhibitor. Ca(2+) and Mg(2+) activated peroxidase while sodium azide, SDS, and Triton X-100 were found to inhibit peroxidase.

  16. The molecular characterization of the lignin-forming peroxidase

    Energy Technology Data Exchange (ETDEWEB)

    Lagrimini, L.M.

    1992-01-01

    This laboratory is committed to understanding the function of plant peroxidases via a multi-disciplinary approach. We have chosen the lignin-forming peroxidase from tobacco as the first isoenzyme to be subjected to this comprehensive approach. The goals which were set out upon the initiation of this project were as follows: (1) utilize a cDNA clone to the tobacco anionic peroxidase to generate transgenic plants which either over-produced this isoenzyme or specifically under-produced this isoenzyme via antisense RNA, (2) describe any phenotypic changes resulting from altered peroxidase expression, (3) perform morphological, physiological, and biochemical analysis of the above mentioned plants to help in determining the in planta function for this enzyme, and (4) clone and characterize the gene for the tobacco anionic peroxidase. A summary of progress thus far which includes both published and unpublished work will be presented in three sections: generation and characterization of transgenic plants, description of phenotypes, and biochemical and physiological analysis of peroxidase function, and cloning and characterization of the tobacco anionic peroxidase gene.

  17. Limits of Versatility of Versatile Peroxidase

    Science.gov (United States)

    Knop, Doriv; Levinson, Dana; Makovitzki, Arik; Agami, Avi; Lerer, Elad; Mimran, Avishai; Yarden, Oded

    2016-01-01

    ABSTRACT Although Mn2+ is the most abundant substrate of versatile peroxidases (VPs), repression of Pleurotus ostreatus vp1 expression occurred in Mn2+-sufficient medium. This seems to be a biological contradiction. The aim of this study was to explore the mechanism of direct oxidation by VP1 under Mn2+-deficient conditions, as it was found to be the predominant enzyme during fungal growth in the presence of synthetic and natural substrates. The native VP1 was purified and characterized using three substrates, Mn2+, Orange II (OII), and Reactive Black 5 (RB5), each oxidized by a different active site in the enzyme. While the pH optimum for Mn2+ oxidation is 5, the optimum pH for direct oxidation of both dyes was found to be 3. Indeed, effective in vivo decolorization occurred in media without addition of Mn2+ only under acidic conditions. We have determined that Mn2+ inhibits in vitro the direct oxidation of both OII and RB5 while RB5 stabilizes both Mn2+ and OII oxidation. Furthermore, OII was found to inhibit the oxidation of both Mn2+ and RB5. In addition, we could demonstrate that VP1 can cleave OII in two different modes. Under Mn2+-mediated oxidation conditions, VP1 was able to cleave the azo bond only in asymmetric mode, while under the optimum conditions for direct oxidation (absence of Mn2+ at pH 3) both symmetric and asymmetric cleavages occurred. We concluded that the oxidation mechanism of aromatic compounds by VP1 is controlled by Mn2+ and pH levels both in the growth medium and in the reaction mixture. IMPORTANCE VP1 is a member of the ligninolytic heme peroxidase gene family of the white rot fungus Pleurotus ostreatus and plays a fundamental role in biodegradation. This enzyme exhibits a versatile nature, as it can oxidize different substrates under altered environmental conditions. VPs are highly interesting enzymes due to the fact that they contain unique active sites that are responsible for direct oxidation of various aromatic compounds

  18. Gene Targeting Without DSB Induction Is Inefficient in Barley.

    Science.gov (United States)

    Horvath, Mihaly; Steinbiss, Hans-Henning; Reiss, Bernd

    2016-01-01

    Double strand-break (DSB) induction allowed efficient gene targeting in barley (Hordeum vulgare), but little is known about efficiencies in its absence. To obtain such data, an assay system based on the acetolactate synthase (ALS) gene was established, a target gene which had been used previously in rice and Arabidopsis thaliana. Expression of recombinases RAD51 and RAD54 had been shown to improve gene targeting in A. thaliana and positive-negative (P-N) selection allows the routine production of targeted mutants without DSB induction in rice. We implemented these approaches in barley and analysed gene targeting with the ALS gene in wild type and RAD51 and RAD54 transgenic lines. In addition, P-N selection was tested. In contrast to the high gene targeting efficiencies obtained in the absence of DSB induction in A. thaliana or rice, not one single gene targeting event was obtained in barley. These data suggest that gene targeting efficiencies are very low in barley and can substantially differ between different plants, even at the same target locus. They also suggest that the amount of labour and time would become unreasonably high to use these methods as a tool in routine applications. This is particularly true since DSB induction offers efficient alternatives. Barley, unlike rice and A. thaliana has a large, complex genome, suggesting that genome size or complexity could be the reason for the low efficiencies. We discuss to what extent transformation methods, genome size or genome complexity could contribute to the striking differences in the gene targeting efficiencies between barley, rice and A. thaliana.

  19. Effects of metoclopramide on mRNA levels of steroid 5α-reductase isozymes in prostate of adult rats.

    Science.gov (United States)

    Sánchez, Pilar; Torres, Jesús M; Castro, Beatriz; Frías, José F; Ortega, Esperanza

    2013-03-01

    The rising incidence of prostate cancer and benign prostatic hypertrophy in the Western world is a cause of increasing public health concern. The most active androgen in the prostate is 5α-dihydrotestosterone obtained from testosterone (T) by the enzyme 5α-reductase (5α-R), expressed in the prostate as two isozymes, 5α-R1 and 5α-R2. These isozymes are involved in the growth and development of normal prostate and in the onset and progression of prostate disease. Besides androgens, prolactin (PRL) may also play a role, although it is not clear whether its effects on the prostate are in synergism with or independent of those of androgens. We previously demonstrated that sulpiride, an inductor of hyperprolactinemia, increased mRNA levels of 5α-R isozymes in prostate of adult rat. We hypothesized a possible interrelationship between PRL levels and 5α-R, although the effects of sulpiride per se cannot be ruled out. In the present study, one-step quantitative reverse transcription polymerase chain reaction coupled with laser-induced fluorescence capillary electrophoresis was used to quantify mRNA levels of both 5α-R isozymes in prostate of adult rat after administration of metoclopramide (MTC), another inductor of PRL secretion. With the administration regimens studied, MTC produced an increase in prostate weight and mRNA levels of 5α-R1 and 5α-R2 in adult rats. Given our finding that MTC per se or MTC-induced hyperprolactinemia modifies prostate disease-related parameters in animals with reduced plasma T levels, further investigation is warranted into the possibility that MTC use by aging males may increase their risk of prostate disease.

  20. Stomach Chitinase from Japanese Sardine Sardinops melanostictus: Purification, Characterization, and Molecular Cloning of Chitinase Isozymes with a Long Linker.

    Science.gov (United States)

    Kawashima, Satoshi; Ikehata, Hiroki; Tada, Chihiro; Ogino, Tomohiro; Kakizaki, Hiromi; Ikeda, Mana; Fukushima, Hideto; Matsumiya, Masahiro

    2016-01-20

    Fish express two different chitinases, acidic fish chitinase-1 (AFCase-1) and acidic fish chitinase-2 (AFCase-2), in the stomach. AFCase-1 and AFCase-2 have different degradation patterns, as fish efficiently degrade chitin ingested as food. For a comparison with the enzymatic properties and the primary structures of chitinase isozymes obtained previously from the stomach of demersal fish, in this study, we purified chitinase isozymes from the stomach of Japanese sardine Sardinops melanostictus, a surface fish that feeds on plankton, characterized the properties of these isozymes, and cloned the cDNAs encoding chitinases. We also predicted 3D structure models using the primary structures of S. melanostictus stomach chitinases. Two chitinase isozymes, SmeChiA (45 kDa) and SmeChiB (56 kDa), were purified from the stomach of S. melanostictus. Moreover, two cDNAs, SmeChi-1 encoding SmeChiA, and SmeChi-2 encoding SmeChiB were cloned. The linker regions of the deduced amino acid sequences of SmeChi-1 and SmeChi-2 (SmeChi-1 and SmeChi-2) are the longest among the fish stomach chitinases. In the cleavage pattern groups toward short substrates and the phylogenetic tree analysis, SmeChi-1 and SmeChi-2 were classified into AFCase-1 and AFCase-2, respectively. SmeChi-1 and SmeChi-2 had catalytic domains that consisted of a TIM-barrel (β/α)₈-fold structure and a deep substrate-binding cleft. This is the first study showing the 3D structure models of fish stomach chitinases.

  1. Purification and properties of three NAD(P)+ isozymes of L-glutamate dehydrogenase of Chlamydomonas reinhardtii.

    Science.gov (United States)

    Moyano, E; Cárdenas, J; Muñoz-Blanco, J

    1992-02-13

    Three isozymes of glutamate dehydrogenase (GDH) of Chlamydomonas reinhardtii, induced under different trophic and stress conditions, have been purified about 800-1000-fold to electrophoretic homogeneity. They are hexamers of Mr 266,000-269,000 as deduced from gel filtration and sedimentation coefficient data. GDH1 consisted of six identical subunits of 44 kDa each, whereas both GDH2 and GDH3 consisted of six similar-sized monomers (4 of 44 kDa and 2 of 46 kDa). Optimum pH for the three activities with each pyridine nucleotide was identical (8.5 with NADH; 7.7 with NADPH; and 9.0 with NAD+). The isozymes exhibited similar high optimum temperature values (60-62 degrees C) and isoelectric points (7.9-8.1). Activity was enhanced in vitro by Ca2+ ions and strongly inhibited by pyridoxal 5'-phosphate, KCN, o-phenanthroline and EDTA, and to a lesser extent by pHMB and methylacetimidate. In the aminating reaction the three isozymes were inhibited in a concentration-dependent process by both NADH and NADPH, with apparent Km values for NH4+ ranging from 13-53 mM; 0.36-1.85 mM for 2-oxoglutarate and 0.07-0.78 mM for NADH and NADPH. In the deaminating reaction apparent Km values ranged from 0.64-3.52 mM for L-glutamate and 0.20-0.32 for NAD+. In addition, the three isozymes exhibited a non-hyperbolic kinetics for NAD+ with negative cooperativity (n = 0.8).

  2. Trypanosoma cruzi: expression of antigenic component 5 among 35 laboratory clones obtained from 18 different isozymic variants

    Directory of Open Access Journals (Sweden)

    Simone F. Breniere

    1987-04-01

    Full Text Available Two monoclonal antibodies anti-component 5 of Trypanosoma cruzi (I-35/115 and II-190/30 were tested in IFA and ELISA respectively against 35 T. cruzi laboratory clones. Among the 35 clones tested, 18 different isozyme patterns were detected. All clones were recognized by both monoclonal antibodies except one clone which did not react with II-190/30. These results support the universal expression of specific component 5 within the taxon T. cruzi.

  3. PHYSIOLOGICAL AND AGROECOLOGICAL ASPECTS OF CADMIUM INTERACTIONS WITH BARLEY PLANTS: AN OVERVIEW

    Directory of Open Access Journals (Sweden)

    A VASSILEV

    2003-07-01

    Full Text Available This work is a review of author’s previous publications, unpublished results as well as available literature on barley responses to Cd contamination. The physiological backgrounds of the acute Cd toxicity in barley plants are briefly described. Some data characterizing the chronic Cd toxicity in barley have been also provided in relation to its possible use for seed production and Cd phytoextraction on Cd-contaminated agricultural soils. Information about the main physiological factors limiting growth of Cd-exposed barley plants and grain yield, seedling quality as well as Cd phytoextraction capacity of barley grown in Cd-contaminated soils is presented.

  4. Identification and characterization of barley RNA-directed RNA polymerases

    DEFF Research Database (Denmark)

    Madsen, Christian Toft; Stephens, Jennifer; Hornyik, Csaba

    2009-01-01

    in dicot species. In this report, we identi!ed and characterized HvRDR1, HvRDR2 and HvRDR6 genes in the monocot plant barley (Hordeum vulgare). We analysed their expression under various biotic and abiotic stresses including fungal and viral infections, salicylic acid treatment as well as during plant...... development. The different classes and subclasses of barley RDRs displayed contrasting expression patterns during pathogen challenge and development suggesting their involvement in speci!c regulatory pathways. Their response to heat and salicylic acid treatment suggests a conserved pattern of expression...

  5. Occurrence of barley leaf disease and control strategies in Denmark

    DEFF Research Database (Denmark)

    Jørgensen, Lise Nistrup; Ørum, Jens Erik; Heick, Thies Marten

    Barley (Hordeum vulgare) is one of the major crops in Denmark and of special importance for malting and for pig feed. In 2016, the crop was grown covering a total area of 700,000 ha; approximately 25% of arable area in Denmark. To ensure high yield of around 60 dt ha-1, disease-tolerant cultivars......), mildew of barley (Erysiphe graminis f.sp. hordei) and Ramularia (Ramularia collo-cygni). In recent years, brown rust and net blotch have been the most important disease in terms of yield losses. As most cultivars have mlo resistance, powdery mildew is today seen as a minor problem. Significant attack...

  6. Biotin Carboxyl Carrier Protein in Barley Chloroplast Membranes

    DEFF Research Database (Denmark)

    Kannangara, C. G.; Jense, C J

    1975-01-01

    Biotin localized in barley chloroplast lamellae is covalently bound to a single protein with an approximate molecular weight of 21000. It contains one mole of biotin per mole of protein and functions as a carboxyl carrier in the acetyl-CoA carboxylase reaction. The protein was obtained by solubil......Biotin localized in barley chloroplast lamellae is covalently bound to a single protein with an approximate molecular weight of 21000. It contains one mole of biotin per mole of protein and functions as a carboxyl carrier in the acetyl-CoA carboxylase reaction. The protein was obtained...

  7. Barley starch bioengineering for high phosphate and amylose

    DEFF Research Database (Denmark)

    Blennow, Per Gunnar Andreas; Carciofi, Massimiliano; Shaik, Shahnoor Sultana

    2011-01-01

    of the three genes encoding the starch-branching enzymes SBEI, SBEIIa, and SBEIIb using a triple RNAi chimeric hairpin construct we generated a virtually amylopectin-free barley. The grains of the transgenic lines were shrunken and had a yield of around 80% of the control line. The starch granules were...... irregular and showed no distinct melting enthalpy and very weak X-ray scattering. Hyperphosphorylated barley starch was achieved by endosperm specific overexpression of the potato glucan water dikinase1 (StGWD1). The content of phosphate esters in this starch was tenfold higher than the control lines...

  8. Implementation of biochemical screening to improve baking quality of Barley

    DEFF Research Database (Denmark)

    Aaslo, Per; Langkilde, Ane; Dionisio, Giuseppe

    2011-01-01

    Barley (Hordeum vulgare) is mostly used in feed and malt production but has the ability to provide humans nutritional benefits. The current wheat based “barley” breads can unfortunately not exceed more than 20% barley flour mixed into the dough due to poor leavening properties. Therefore...... the opportunity to give a forecast of the taste of the bread, as the AA composition is known to control certain aspects of the taste. We uses a MSE approach on a time of flight instrument coupled to a UPLC and in gel digestion to identify and characterize the different D-hordeins responsible for baking quality...

  9. The transfer of {sup 137}Cs from barley to beer

    Energy Technology Data Exchange (ETDEWEB)

    Proehl, G.; Mueller, H.; Voigt, G. [Institut fuer Strahlenschutz, Oberschleibheim (Germany)] [and others

    1997-01-01

    Beer has been brewed from barley contaminated with {sup 137}Cs as a consequence of the Chernobyl accident. The {sup 137}Cs activity has been measured in all intermediate steps and in the by-products of the production process. About 35 % of the {sup 137}Cs in barley were recovered in beer. Processing factors defined as the concentration ratio of processed and raw products were determined to be 0.61, 3.3, 0.1 and 0.11 for malt, malt germs, spent grains and beer, respectively. 4 refs., 2 tabs.

  10. Bisphenol A Modifies the Regulation Exerted by Testosterone on 5α-Reductase Isozymes in Ventral Prostate of Adult Rats

    Directory of Open Access Journals (Sweden)

    Pilar Sánchez

    2013-01-01

    Full Text Available The development, growth, and function of the prostate gland depend on androgen stimulation. The primary androgen in prostate is 5-dihydrotestosterone (DHT which is synthesized from circulating testosterone (T through the action of 5-reductase (5-R. Although 5-R occurs as five isozymes, only 5-R1 and 5-R2 are physiologically involved in steroidogenesis. The endocrine disruptor bisphenol A (BPA alters sexual organs, including the prostate. Our previous findings indicated that BPA decreased the expression of 5-R1 and 5-R2 in rat prostate but also circulating T. Thus, it is unclear whether BPA exerts this effect on 5-R isozymes by reducing circulating T or by any other mechanism. In this study, we examine the effects of short-term exposure to BPA at doses below 25 g/Kg/d and above 300 g/Kg/d of the TDI on mRNA levels of 5-R1 and 5-R2 in prostate of adult castrated rats supplemented with T to achieve constant circulating T levels. mRNA levels were measured by absolute quantitative RT-PCR, T levels by RIA, and DHT levels by ELISA. Our results indicated that in castrated rats treated with T BPA at the two doses studied significantly decreased the mRNA levels of both 5-R isozymes in a dose-dependent manner without modifications in circulating T.

  11. Changes of serum amylase, its isozyme fractions and amylase-creatinine clearance ratio in dogs with experimentally induced acute pancreatitis.

    Science.gov (United States)

    Akuzawa, M; Morizono, M; Nagata, K; Hayano, S; Sakamoto, H; Yasuda, N; Okamoto, K; Kawasaki, Y; Deguchi, E

    1994-04-01

    To investigate the diagnostic application of amylase to canine pancreatic diseases, serum amylase activities, its isozyme fractions and amylase-creatinine clearance ratio (ACCR) were analyzed in normal intact dogs and dogs experimentally induced acute pancreatitis. There was no statistic difference between normal male and female dogs. Amylase specific activities in pancreatic tissue extracts were more than 2,300 times higher than that in serum, and were also higher than those in other tissues; parotid and mandibular salivary glands, lung, heart, liver, spleen, duodenum, jejunum, ileum and kidney. Following the chloroform injection into the pancreatic tissue, WBC increased from 6 to 240 hr and serum glucose significantly increased at 72 and 96 hr, and no urine glucose was detected. BUN as well as serum and urine creatinine showed normal levels. ACCR increased until 96 hr without statistic significance. Serum amylase activities increased significantly after 3 hr and its isozyme was separated into 4 fractions (Amy1-Amy4) in contrast to 3 fractions (Amy2-Amy4) in intact dogs. Since this extra Amy1 seen from 1 hr increasing after 6 hr similarly to other 3 fractions, the evaluation of serum amylase and its isozyme fractions was indicated to be useful for the diagnosis of acute pancreatitis in dogs.

  12. Diversity evaluation based on morphological, physiological and isozyme variation in genetic resources of garlic (Allium sativum L.) collected worldwide.

    Science.gov (United States)

    Hirata, Sho; Abdelrahman, Mostafa; Yamauchi, Naoki; Shigyo, Masayoshi

    2016-11-26

    The aim of this study was to obtain primary information about the global diversity of garlic (Allium sativum L.) by evaluating morphological, physiological and isozyme variation. A total of 107 garlic accessions collected worldwide were grown in Yamaguchi, Japan. Five morphological traits (bulb weight, bulb diameter, number of cloves per bulb, number of bulbils and scape length) and one physiological trait (bolting period) of the collected garlic showed wide variation. Meanwhile, a total of 140 garlic accessions, including the 107 mentioned above, were characterized by leucine aminopeptidase (LAP) and phosphoglucoisomerase (PGI) isozyme analyses; they clearly showed polymorphisms in putative isozyme loci (Lap-1, Lap-2 and Pgi-1). Allelic frequencies were estimated in each group of accessions categorized by their geographical origin, and the observed (Ho) and expected (He) heterozygosities were calculated. The allelic frequencies differed between groups. A principal component analysis based on morpho-physiological data indicated a grouping of the garlic accessions into Central Asian and Northern Mediterranean groups as well as others. We discuss the roles of artificial and natural selection that may have caused differentiation in these traits, on the assumption that ancestral domesticated garlic populations have adapted in various regions using standing variation or mutations that accumulated during expansion, and have evolved along with human-preferred traits over a long history of cultivation.

  13. Consistency of population genetics parameters estimated from isozyme and RAPDs dataset in species of genus Prosopis (Leguminosae, Mimosoideae).

    Science.gov (United States)

    Ferreyra, Laura Inés; Bessega, Cecilia; Vilardi, Juan C; Saidman, Beatriz O

    2007-11-01

    Genetic variability, population structure and differentiation among 17 populations of 5 species and 2 natural interspecific hybrids of section Algarobia of genus Prosopis were analyzed from data of 23 isozyme and 28 RAPD loci. Both markers indicated that the studied populations are highly variable. P. alba populations in average showed lower values of genetic variability estimates from isozyme data, but this trend was not observed for RAPD markers. The hierarchical analyses of the distribution of genetic variability showed that the highest proportion of variation occurred within populations, the differentiation among species was intermediate and the lowest component was observed among populations within species. The consistency between results from both dataset implies that they are not biased and reflect the actual genetic structure of the populations analyzed. The matrices of Euclidean distances obtained from the two sets of markers were highly correlated according to Mantel test. In both cases the corresponding phenogram and MDS plot tended to cluster conspecific populations while hybrid populations were not intermediate between putative parents. Some disagreements between isozyme and RAPD phenograms were observed mainly in the affinities of hybrid populations. Such inconsistencies might result from reticular rather than dichotomic evolutionary relationships. The phenetic associations retrieved gave no support to the division of the section Algarobia into series.

  14. Arabidopsis GERANYLGERANYL DIPHOSPHATE SYNTHASE 11 is a hub isozyme required for the production of most photosynthesis-related isoprenoids.

    Science.gov (United States)

    Ruiz-Sola, M Águila; Coman, Diana; Beck, Gilles; Barja, M Victoria; Colinas, Maite; Graf, Alexander; Welsch, Ralf; Rütimann, Philipp; Bühlmann, Peter; Bigler, Laurent; Gruissem, Wilhelm; Rodríguez-Concepción, Manuel; Vranová, Eva

    2016-01-01

    Most plastid isoprenoids, including photosynthesis-related metabolites such as carotenoids and the side chain of chlorophylls, tocopherols (vitamin E), phylloquinones (vitamin K), and plastoquinones, derive from geranylgeranyl diphosphate (GGPP) synthesized by GGPP synthase (GGPPS) enzymes. Seven out of 10 functional GGPPS isozymes in Arabidopsis thaliana reside in plastids. We aimed to address the function of different GGPPS paralogues for plastid isoprenoid biosynthesis. We constructed a gene co-expression network (GCN) using GGPPS paralogues as guide genes and genes from the upstream and downstream pathways as query genes. Furthermore, knock-out and/or knock-down ggpps mutants were generated and their growth and metabolic phenotypes were analyzed. Also, interacting protein partners of GGPPS11 were searched for. Our data showed that GGPPS11, encoding the only plastid isozyme essential for plant development, functions as a hub gene among GGPPS paralogues and is required for the production of all major groups of plastid isoprenoids. Furthermore, we showed that the GGPPS11 protein physically interacts with enzymes that use GGPP for the production of carotenoids, chlorophylls, tocopherols, phylloquinone, and plastoquinone. GGPPS11 is a hub isozyme required for the production of most photosynthesis-related isoprenoids. Both gene co-expression and protein-protein interaction likely contribute to the channeling of GGPP by GGPPS11. © 2015 The Authors. New Phytologist © 2015 New Phytologist Trust.

  15. Arginase deficiency manifesting delayed clinical sequelae and induction of a kidney arginase isozyme.

    Science.gov (United States)

    Grody, W W; Kern, R M; Klein, D; Dodson, A E; Wissman, P B; Barsky, S H; Cederbaum, S D

    1993-03-01

    Deficiency of liver arginase (AI) is characterized clinically by hyperargininemia, progressive mental impairment, growth retardation, spasticity, and periodic episodes of hyperammonemia. The rarest of the inborn errors of urea cycle enzymes, it has been considered the least life-threatening, by virtue of the typical absence of catastrophic neonatal hyperammonemia and its compatibility with a longer life span. This has been attributed to the persistence of some ureagenesis in these patients through the activity of a second isozyme of arginase (AII) located predominantly in the kidney. We have treated a number of arginase-deficient patients into young adulthood. While they are severely retarded and wheelchair-bound, their general medical care has been quite tractable. Recently, however, two of the oldest (M.U., age 20, and M.O., age 22) underwent rapid deterioration, ending in hyperammonemic coma and death, precipitated by relatively minor viral respiratory illnesses inducing a catabolic state with increased endogenous nitrogen load. In both cases, postmortem examination revealed severe global cerebral edema and aspiration pneumonia. Enzyme assays confirmed the absence of AI activity in the livers of both patients. In contrast, AII activity (identified by its different cation cofactor requirements and lack of precipitation with anti-AI antibody) was markedly elevated in kidney tissues, 20-fold in M.O. and 34-fold in M.U. Terminal plasma arginine (1500 mumols/l) and ammonia (1693 mmol/l) levels of M.U. were substantially higher than those of M.O. (348 mumols/l and 259 mumols/l, respectively). By Northern blot analysis, AI mRNA was detected in M.O.'s liver but not in M.U.'s; similarly, anti-AI crossreacting material was observed by Western blot in M.O. only. These findings indicate that, despite their more long-lived course, patients with arginase deficiency remain vulnerable to the same catastrophic events of hyperammonemia that patients with other urea cycle

  16. Peroxidase gene expression during tomato fruit ripening

    Energy Technology Data Exchange (ETDEWEB)

    Biggs, M.S.; Flurkey, W.H.; Handa, A.K.

    1987-04-01

    Auxin oxidation has been reported to play a critical role in the initiation of pear fruit ripening and a tomato fruit peroxidase (POD) has been shown to have IAA-oxidase activity. However, little is known about changes in the expression of POD mRNA in tomato fruit development. They are investigating the expression of POD mRNA during tomato fruit maturation. Fruit pericarp tissues from six stages of fruit development and ripening (immature green, mature green, breaker, turning, ripe, and red ripe fruits) were used to extract poly (A)/sup +/ RNAs. These RNAs were translated in vitro in a rabbit reticulocyte lysate system using L-/sup 35/S-methionine. The /sup 35/S-labeled products were immunoprecipitated with POD antibodies to determine the relative proportions of POD mRNA. High levels of POD mRNA were present in immature green and mature green pericarp, but declined greatly by the turning stage of fruit ripening. In addition, the distribution of POD mRNA on free vs bound polyribosomes will be presented, as well as the presence or absence of POD mRNA in other tomato tissues.

  17. Immobilization of horseradish peroxidase onto kaolin.

    Science.gov (United States)

    Šekuljica, Nataša Ž; Prlainović, Nevena Ž; Jovanović, Jelena R; Stefanović, Andrea B; Djokić, Veljko R; Mijin, Dušan Ž; Knežević-Jugović, Zorica D

    2016-03-01

    Kaolin showed as a very perspective carrier for the enzyme immobilization and it was used for the adsorption of horseradish peroxidase (HRP). The effects of the enzyme concentration and pH on the immobilization efficiency were studied in the reaction with pyrogallol and anthraquinone dye C.I. Acid Violet 109 (AV 109). In addition, Fourier transform infrared spectroscopy, scanning electron microscopy and analysis by Brunauer-Emmett-Teller were performed for kaolin, thermally activated kaolin and the immobilized enzyme. It has been shown that 0.1 IU of HRP-kaolin decolorized 87 % of dye solution, under the optimal conditions (pH 5.0, temperature 24 °C, dye concentration 40 mg/L and 0.2 mM of H2O2) within 40 min. The immobilized HRP decolorization follows the Ping Pong Bi-Bi mechanism with dead-end inhibition by the dye. The biocatalyst retained 35 ± 0.9 % of the initial activity after seven cycles of reuse in the decolorization reaction of AV 109 under optimal conditions in a batch reactor. The obtained kinetic parameters and reusability study confirmed improvement in performances of k-HRP compared to free, indicating that k-HRP has a great potential for environmental purposes.

  18. Class III peroxidases in plant defence reactions.

    Science.gov (United States)

    Almagro, L; Gómez Ros, L V; Belchi-Navarro, S; Bru, R; Ros Barceló, A; Pedreño, M A

    2009-01-01

    When plants are attacked by pathogens, they defend themselves with an arsenal of defence mechanisms, both passive and active. The active defence responses, which require de novo protein synthesis, are regulated through a complex and interconnected network of signalling pathways that mainly involve three molecules, salicylic acid (SA), jasmonic acid (JA), and ethylene (ET), and which results in the synthesis of pathogenesis-related (PR) proteins. Microbe or elicitor-induced signal transduction pathways lead to (i) the reinforcement of cell walls and lignification, (ii) the production of antimicrobial metabolites (phytoalexins), and (iii) the production of reactive oxygen species (ROS) and reactive nitrogen species (RNS). Among the proteins induced during the host plant defence, class III plant peroxidases (EC 1.11.1.7; hydrogen donor: H(2)O(2) oxidoreductase, Prxs) are well known. They belong to a large multigene family, and participate in a broad range of physiological processes, such as lignin and suberin formation, cross-linking of cell wall components, and synthesis of phytoalexins, or participate in the metabolism of ROS and RNS, both switching on the hypersensitive response (HR), a form of programmed host cell death at the infection site associated with limited pathogen development. The present review focuses on these plant defence reactions in which Prxs are directly or indirectly involved, and ends with the signalling pathways, which regulate Prx gene expression during plant defence. How they are integrated within the complex network of defence responses of any host plant cell will be the cornerstone of future research.

  19. Carbon Nanodots as Peroxidase Nanozymes for Biosensing

    Directory of Open Access Journals (Sweden)

    Bhaskar Garg

    2016-12-01

    Full Text Available ‘Nanozymes’, a term coined by Scrimin, Pasquato, and co-workers to describe nanomaterials with enzyme-like characteristics, represent an exciting and emerging research area in the field of artificial enzymes. Indubitably, the last decade has witnessed substantial advancements in the design of a variety of functional nanoscale materials, including metal oxides and carbon-based nanomaterials, which mimic the structures and functions of naturally occurring enzymes. Among these, carbon nanodots (C-dots or carbon quantum dots (CQDs offer huge potential due to their unique properties as compared to natural enzymes and/or classical artificial enzymes. In this mini review, we discuss the peroxidase-like catalytic activities of C-dots and their applications in biosensing. The scope intends to cover not only the C-dots but also graphene quantum dots (GQDs, doped C-dots/GQDs, carbon nitride dots, and C-dots/GQDs nanocomposites. Nevertheless, this mini review is designed to be illustrative, not comprehensive.

  20. Redundancy among manganese peroxidases in Pleurotus ostreatus.

    Science.gov (United States)

    Salame, Tomer M; Knop, Doriv; Levinson, Dana; Yarden, Oded; Hadar, Yitzhak

    2013-04-01

    Manganese peroxidases (MnPs) are key players in the ligninolytic system of white rot fungi. In Pleurotus ostreatus (the oyster mushroom) these enzymes are encoded by a gene family comprising nine members, mnp1 to -9 (mnp genes). Mn(2+) amendment to P. ostreatus cultures results in enhanced degradation of recalcitrant compounds (such as the azo dye orange II) and lignin. In Mn(2+)-amended glucose-peptone medium, mnp3, mnp4, and mnp9 were the most highly expressed mnp genes. After 7 days of incubation, the time point at which the greatest capacity for orange II decolorization was observed, mnp3 expression and the presence of MnP3 in the extracellular culture fluids were predominant. To determine the significance of MnP3 for ligninolytic functionality in Mn(2+)-sufficient cultures, mnp3 was inactivated via the Δku80 strain-based P. ostreatus gene-targeting system. In Mn(2+)-sufficient medium, inactivation of mnp3 did not significantly affect expression of nontargeted MnPs or their genes, nor did it considerably diminish the fungal Mn(2+)-mediated orange II decolorization capacity, despite the significant reduction in total MnP activity. Similarly, inactivation of either mnp4 or mnp9 did not affect orange II decolorization ability. These results indicate functional redundancy within the P. ostreatus MnP gene family, enabling compensation upon deficiency of one of its members.

  1. Antioxidant and isozyme features of two strains of Laminaria japonica (Phaeophyceae)

    Science.gov (United States)

    Wang, You; Tang, Xuexi; Li, Yongqi; Yu, Zhiming

    2007-01-01

    Healthy sporophytes of two gametophyte mutants of Laminaria japonica with different heat resistances: kelp 901 ( 901, with comparatively stronger heat-resistance) and Rongcheng No.1 ( RC, sensitive to heat stress), were respectively collected during October to December 2002 from Yantai and Rongcheng Sea Farm in the Shandong Peninsula of China. The contents of some biochemical materials and antioxidant capacity were analyzed under controlled laboratory conditions to identify if there is any relation between the overall antioxidant capacity and the heat-resistance in L. japonica and to understand possible mechanism of heat-resistance. Results show that: (1) the overall antioxidant capacity in healthy sporophyte of 901, such as vitamin E, polyphenol, and ascorbic acid contents and the enzymatic activity of SOD, POD, CAT, Gpx, PPO, and PAL, were not always higher than that of RC under controlled laboratory conditions, and no significance ( P>0.05) was shown in total antioxidant capacity (T-AOC) in 901 and RC. Result suggested that the difference in antioxidant capacity was not a decisive factor for different heat-resistances in L. japonica; (2) the simultaneous assay on isozymes was carried out using vertical polyacrylamide gel electrophoresis (PAGE). Considerable differences in peroxide (PRX), malate dehydrogenase (MDH), malic enzyme (ME), polyphenol oxidase (PPO) and glutamate dehydrogenase (GDH) were obtained in 901 and RC from either the band number, relative mobility ( R f ), or staining intensity, and ME could be used as an indicator to distinguish healthy sporophyte of 901 and RC under controlled laboratory conditions.

  2. Antioxidant and isozyme features of two strains of Laminaria japonica (Phaeophyceae)

    Institute of Scientific and Technical Information of China (English)

    WANG You; TANG Xuexi; LI Yongqi; YU Zhiming

    2007-01-01

    Healthy sporophytes of two gametophyte mutants of Laminariajaponica with different heat resistances: kelp 901 (901, with comparatively stronger heat-resistance) and Rongcheng No. 1 (RC, sensitive to heat stress), were respectively collected during October to December 2002 from Yantai and Rongcheng Sea Farm in the Shandong Peninsula of China. The contents of some biochemical materials and antioxidant capacity were analyzed under controlled laboratory conditions to identify if there is any relation between the overall antioxidant capacity and the heat-resistance in L. japonica and to understand possible mechanism of heat-resistance. Results show that: (1) the overall antioxidant capacity in healthy sporophyte of 901, such as vitamin E, polyphenol, and ascorbic acid contents and the enzymatic activity of SOD, POD, CAT, Gpx, PPO, and PAL, were not always higher than that of RC under controlled laboratory conditions, and no significance (P>0.05) was shown in total antioxidant capacity (T-AOC) in 901and RC. Result suggested that the difference in antioxidant capacity was not a decisive factor for different heat-resistances in L. japonica; (2) the simultaneous assay on isozymes was carried out using vertical polyacrylamide gel electrophoresis (PAGE). Considerable differences in peroxide (PRX), malate dehydrogenase (MDH), malic enzyme (ME), polyphenol oxidase (PPO) and glutamate dehydrogenase (GDH)were obtained in 901 and RC from either the band number, relative mobility (Rf), or staining intensity, and ME could be used as an indicator to distinguish healthy sporophyte of 901 and RC under controlled laboratory conditions.

  3. Esterase isozymes patterns of grape vine (Vitis vinifera L. are altered in response to fungicide exposure

    Directory of Open Access Journals (Sweden)

    Gleice Ribeiro Orasmo

    2015-10-01

    Full Text Available Current analysis characterizes the effect of different fungicides often applied for pest control on a-and b-esterase patterns of four economically important table-wine grape cultivars (Italia, Rubi, Benitaka and Brasil of Vitis vinifera. The a- and b-esterase patterns in bud leaves of the cultivars were assessed by native PAGE analysis. Cabrio Top® compound inhibited Est-2, Est-5, Est-6, Est-7, Est-8, Est-9 and Est-10 carboxylesterases, whereas Est-4, Est-11, Est-12, Est-13, Est-14 acetylesterases and Est-16 carboxylesterase were detected as weakly stained bands. Carboxylesterases and acetylesterases were also detected as weakly stained bands when exposed to fungicides Orthocide 500®, Positron Duo® and Folicur PM®. No changes in a- and b-esterase patterns were reported when the vines were exposed to the fungicides Rovral SC®, Kumulus DF®, Curzate M®, Score® or Cuprogarb 500®. The evidence of functional changes in carboxylesterase and acetylesterase levels in current study is a warning to grape producers on the dangers inherent in the indiscriminate use of potent and modern fungicides extensively used in agriculture. The inhibition effect of fungicides on esterase isozyme molecules seems to be independent of the fungicide chemical.

  4. Auto-inhibition and phosphorylation-induced activation of PLC-γ isozymes

    Science.gov (United States)

    Hajicek, Nicole; Charpentier, Thomas H.; Rush, Jeremy R.; Harden, T. Kendall; Sondek, John

    2013-01-01

    Multiple extracellular stimuli, such as growth factors and antigens, initiate signaling cascades through tyrosine phosphorylation and activation of phospholipase C (PLC)-γ isozymes. Like most other PLCs, PLC-γ1 is basally auto-inhibited by its X-Y linker, which separates the X-and Y-boxes of the catalytic core. The C-terminal SH2 (cSH2) domain within the X-Y linker is the critical determinant for auto-inhibition of phospholipase activity. Release of auto-inhibition requires an intramolecular interaction between the cSH2 domain and a phosphorylated tyrosine, Tyr783, also located within the X-Y linker. The molecular mechanisms that mediate auto-inhibition and phosphorylation-induced activation have not been defined. Here, we describe structures of the cSH2 domain both alone and bound to a PLC-γ1 peptide encompassing phosphorylated Tyr783. The cSH2 domain remains largely unaltered by peptide engagement. Point mutations in the cSH2 domain located at the interface with the peptide were sufficient to constitutively activate PLC-γ1 suggesting that peptide engagement directly interferes with the capacity of the cSH2 domain to block the lipase active site. This idea is supported by mutations in a complimentary surface of the catalytic core that also enhanced phospholipase activity. PMID:23777354

  5. Structures of starches from rice mutants deficient in the starch synthase isozyme SSI or SSIIIa.

    Science.gov (United States)

    Hanashiro, Isao; Higuchi, Toshiyuki; Aihara, Satomi; Nakamura, Yasunori; Fujita, Naoko

    2011-05-09

    Amylose and amylopectin of rice mutants deficient in a starch synthase (SS) isozyme in the endosperm, either SSI (ΔSSI) or SSIIIa (ΔSSIIIa), were structurally altered from those of their parent (cv. Nipponbare, Np). The amylose content was higher in the mutants (Np, 15.5%; ΔSSI, 18.2%; ΔSSIIIa, 23.6%), and the molar ratio of branched amylose and its side chains was increased. The chain-length distribution of the β-amylase limit dextrins of amylopectin showed regularity, which appeared consistent with the generally accepted cluster structure, and the degrees of polymerization found at the intersections were taken as the boundaries of the B-chain fractions. The mole % of the B(1)-B(3) fractions was changed slightly in ΔSSI, which is consistent with the proposed role of SSI in elongating the external part of clusters. In ΔSSIIIa, a significant increase in the B(1) fraction and a decrease in the B(2) and B(3) fractions were observed. The internal chain length of the B(2) and B(3) fractions appeared to be slightly altered, suggesting that the deficiency in SS affected the actions of branching enzyme(s).

  6. Antibodies reacting to carbonic anhydrase isozymes (I and II) and albumin in sera from dogs.

    Science.gov (United States)

    Nishita, Toshiho; Miyazaki, Rui; Miyazaki, Takae; Ochiai, Hideharu; Orito, Kensuke

    2016-06-01

    IgGs to carbonic anhydrase isozymes (CA-I and CA-II) and albumin were identified in dog serum. IgG titers were determined in the sera of asymptomatic dogs, and in dogs with atopic dermatitis, diarrhea and/or vomiting, diabetes and/or pancreatitis, kidney disease, hepatic disease, and thyroid gland disease, using ELISA. Low titres of IgG-reactive CA-I, CA-II, BSA, and CSA were found in the sera of healthy beagles. Compared with healthy beagles, there was a significant difference in the titers of antibodies against CA-I in asymptomatic dogs, dogs with diabetes and/or pancreatitis, or thyroid gland disease, or hepatic disease. Compared with healthy beagles, there was a significant difference in the antibody titer of anti-CA-II IgG in asymptomatic dogs and in those with hepatic disease. There was a significant difference in the antibody titer of anti-BSA IgG between healthy beagles and dogs with hepatic disease.

  7. Changes in specific activity of ascorbate peroxidase during seed ...

    African Journals Online (AJOL)

    USER

    2010-08-16

    Aug 16, 2010 ... modes of application of SA, it was observed that maximum specific activity of .... One gram seeds were homogenized at 4°C with pestle and mortar .... properties and distribution of ascorbate peroxidase in legume root nodules.

  8. Altered phenotypes in plants transformed with chimeric tobacco peroxidase genes

    Energy Technology Data Exchange (ETDEWEB)

    Lagrimini, L.M.

    1990-12-31

    Peroxidases have been implicated in a variety of secondary metabolic reactions including lignification, cross-linking of cell wall polysaccharides, oxidation of indole-3-acetic acid, regulation of cell elongation, wound-healing, phenol oxidation, and pathogen defense. However, due to the many different isoenzymes and even more potential substrates, it has proven difficult to verify actual physiological roles for peroxidase. We are studying the molecular biology of the tobacco peroxidase genes, and have utilized genetic engineering techniques to produce transgenic plants which differ only in their expression of an individual peroxidase isoenzyme. Many of the in planta functions for any individual isoenzyme may be predicted through the morphological and physiological analysis of transformed plants.

  9. Altered phenotypes in plants transformed with chimeric tobacco peroxidase genes

    Energy Technology Data Exchange (ETDEWEB)

    Lagrimini, L.M.

    1990-01-01

    Peroxidases have been implicated in a variety of secondary metabolic reactions including lignification, cross-linking of cell wall polysaccharides, oxidation of indole-3-acetic acid, regulation of cell elongation, wound-healing, phenol oxidation, and pathogen defense. However, due to the many different isoenzymes and even more potential substrates, it has proven difficult to verify actual physiological roles for peroxidase. We are studying the molecular biology of the tobacco peroxidase genes, and have utilized genetic engineering techniques to produce transgenic plants which differ only in their expression of an individual peroxidase isoenzyme. Many of the in planta functions for any individual isoenzyme may be predicted through the morphological and physiological analysis of transformed plants.

  10. Optimization of extracellular fungal peroxidase production by 2 Coprinus species

    National Research Council Canada - National Science Library

    Keisuke Ikehata; Michael A. Pickard; Ian D. Buchanan; Daniel W. Smith

    2004-01-01

    .... Of the 7 factors examined in the screening study, the concentrations of carbon (glucose) and nitrogen (peptone or casitone) sources showed significant effects on the peroxidase production by Coprinus sp...

  11. Biomimetic Synthesis of Resveratrol Trimers Catalyzed by Horseradish Peroxidase

    National Research Council Canada - National Science Library

    Jian-Qiao Zhang; Gan-Peng Li; Yu-Long Kang; Bin-Hao Teng; Chun-Suo Yao

    2017-01-01

    Biotransformation of trans-resveratrol and synthetic (±)-ε-viniferin in aqueous acetone using horseradish peroxidase and hydrogen peroxide as oxidants resulted in the isolation of two new resveratrol trimers (3 and 4...

  12. Heme electron transfer in peroxidases: the propionate e-pathway.

    Science.gov (United States)

    Guallar, Victor

    2008-10-23

    Computational modeling offers a new insight about the electron transfer pathway in heme peroxidases. Available crystal structures have revealed an intriguing arrangement of the heme propionate side chains in heme-heme and heme-substrate complexes. By means of mixed quantum mechanical/molecular mechanics calculations, we study the involvement of these propionate groups into the substrate oxidation in ascorbate peroxidase and into the heme to heme electron transfer in bacterial cytochrome c peroxidase. By selectively turning on/off different quantum regions, we obtain the electron transfer pathway which directly involves the porphyrin ring and the heme propionates. Furthermore, in ascorbate peroxidase the presence of the substrate appears to be crucial for the activation of the electron transfer channel. The results might represent a general motif for electron transfer from/to the heme group and change our view for the propionate side chains as simple electrostatic binding anchors. We name the new mechanism "the propionate e-pathway".

  13. Cell wall bound anionic peroxidases from asparagus byproducts.

    Science.gov (United States)

    Jaramillo-Carmona, Sara; López, Sergio; Vazquez-Castilla, Sara; Jimenez-Araujo, Ana; Rodriguez-Arcos, Rocio; Guillen-Bejarano, Rafael

    2014-10-08

    Asparagus byproducts are a good source of cationic soluble peroxidases (CAP) useful for the bioremediation of phenol-contaminated wastewaters. In this study, cell wall bound peroxidases (POD) from the same byproducts have been purified and characterized. The covalent forms of POD represent >90% of the total cell wall bound POD. Isoelectric focusing showed that whereas the covalent fraction is constituted primarily by anionic isoenzymes, the ionic fraction is a mixture of anionic, neutral, and cationic isoenzymes. Covalently bound peroxidases were purified by means of ion exchange chromatography and affinity chromatography. In vitro detoxification studies showed that although CAP are more effective for the removal of 4-CP and 2,4-DCP, anionic asparagus peroxidase (AAP) is a better option for the removal of hydroxytyrosol (HT), the main phenol present in olive mill wastewaters.

  14. Kinetic Study on Horseradish Peroxidase Interacting with Cyclodextrin

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    The catalyst reactivity of Horseradish peroxidase was enhanced in the presence of β- cyclodextrin. During this course, β- cyclodextrin played a role of stabilizing the intermediates of HRP. The results have been investigated using spectra and calculation.

  15. The effect of heavy metals on peroxidase from Jerusalem artichoke ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-07-04

    Jul 4, 2008 ... African Journal of Biotechnology Vol. ... EU/mg. The substrate specificity of peroxidase was investigated ... compounds, removal of phenolics from waste waters and ... Heavy metal pollution occurs in many industrial waste-.

  16. Spatio-temporal changes in germination and radical elongation of barley seeds tracked by proteome analysis of dissected embryo, aleurone layer and endosperm tissues

    DEFF Research Database (Denmark)

    Bønsager, Birgit Christine; Finnie, Christine; Roepstorff, P.

    2007-01-01

    Germination of barley is accompanied by changes in water-soluble seed proteins. 2-DE was used to describe spatio-temporal proteome differences in dissected seed tissues associated with germination and the subsequent radicle elongation. Protein identification by MS enabled assignment of proteins...... protein and an ABA-induced protein, suggesting that these proteins are also involved in desiccation stress. Several redox-related proteins differed in spatio-temporal patterns at the end of germination and onset of radicle elongation. Notably, ascorbate peroxidase that was observed only in the embryo......, increased in abundance at 36 h PI. The surprisingly early changes seen in the protein profiles already 4 h after imbibition indicate that germination is programmed during seed maturation...

  17. Modulation of Kv3.1b potassium channel phosphorylation in auditory neurons by conventional and novel protein kinase C isozymes.

    Science.gov (United States)

    Song, Ping; Kaczmarek, Leonard K

    2006-06-02

    In fast-spiking neurons such as those in the medial nucleus of the trapezoid body (MNTB) in the auditory brainstem, Kv3.1 potassium channels are required for high frequency firing. The Kv3.1b splice variant of this channel predominates in the mature nervous system and is a substrate for phosphorylation by protein kinase C (PKC) at Ser-503. In resting neurons, basal phosphorylation at this site decreases Kv3.1 current, reducing neuronal ability to follow high frequency stimulation. We used a phospho-specific antibody to determine which PKC isozymes control serine 503 phosphorylation in Kv3.1b-tranfected cells and in auditory neurons in brainstem slices. By using isozyme-specific inhibitors, we found that the novel PKC-delta isozyme, together with the novel PKC-epsilon and conventional PKCs, contributed to the basal phosphorylation of Kv3.1b in MNTB neurons. In contrast, only PKC-epsilon and conventional PKCs mediate increases in phosphorylation produced by pharmacological activation of PKC in MNTB neurons or by metabotropic glutamate receptor activation in Kv3.1/mGluR1-cotransfected cells. We also measured the time course of dephosphorylation and recovery of basal phosphorylation of Kv3.1b following brief high frequency electrical stimulation of the trapezoid body, and we determined that the recovery process is mediated by both novel PKC-delta and PKC-epsilon isozymes and by conventional PKCs. The association between Kv3.1b and PKC isozymes was confirmed by reciprocal coimmunoprecipitation of Kv3.1b with multiple PKC isozymes. Our results suggest that the Kv3.1b channel is regulated by both conventional and novel PKC isozymes and that novel PKC-delta contributes specifically to the maintenance of basal phosphorylation in auditory neurons.

  18. Ionically Bound Peroxidase from Peach Fruit

    Directory of Open Access Journals (Sweden)

    Neves Valdir Augusto

    2002-01-01

    Full Text Available Soluble, ionically bound peroxidase (POD and polyphenoloxidase (PPO were extracted from the pulp of peach fruit during ripening at 20°C. Ionically bound form was purified 6.1-fold by DEAE-cellulose and Sephadex G-100 chromatography. The purified enzyme showed only one peak of activity on Sephadex G-100 and PAGE revealed that the enzyme was purified by the procedures adopted. The purified enzyme showed a molecular weight of 29000 Da, maximum activity at pH 5.0 and at 40ºC. The calculated apparent activation energy (Ea for the reaction was10.04 kcal/mol. The enzyme was heat-labile in the temperature range of 60 to 75ºC with a fast inactivation at 75ºC. Measurement of residual activity showed a stabilizing effect of sucrose at various temperature/sugar concentrations (0, 10, 20 %, w/w, with an activation energy (Ea for inactivation increasing with sucrose concentration from 0 to 20% (w/w. The Km and Vmax values were 9.35 and 15.38 mM for 0-dianisidine and H2O2, respectively. The bound enzyme was inhibited competitively by ferulic, caffeic and protocatechuic acids with different values of Ki,. L-cysteine, p-coumaric and indolacetic acid and Fe++ also inhibited the enzyme but at a lower grade. N-ethylmaleimide and p-CMB were not effective to inhibit the enzyme demonstrating the non-essentiality of SH groups.

  19. Wheat and barley exposure to nanoceria: Implications for agricultural productivity

    Science.gov (United States)

    The impacts of man-made nanomaterials on agricultural productivity are not yet well understood. A soil microcosm study was performed to assess the physiological, phenological, and yield responses of wheat (Triticum aestivum) and barley (Hordeum vulgare L.) exposed to nanoceria (n...

  20. Classification and salt tolerance analysis of barley varieties

    NARCIS (Netherlands)

    Katerji, N.; Hoorn, van J.W.; Hamdy, A.; Mastrorilli, M.; Fares, C.; Ceccarelli, S.; Grando, S.; Oweis, T.

    2006-01-01

    Six varieties of barley (Hordeum vulgare), five of which were provided by ICARDA, were tested in a green house experiment for their salt tolerance. Afterwards the ICARDA variety Melusine, selected from this experiment for its combination of high yield and salt tolerance, was compared in a lysimeter

  1. Durum wheat and barley productivity in saline-drought environments

    NARCIS (Netherlands)

    Katerji, N.; Mastrorilli, M.; Hoorn, van J.W.; Lahmer, F.Z.; Hamdy, A.; Oweis, T.

    2009-01-01

    In two Successive years, durum wheat (Triticum turgidum Desf.) and barley (Hodeum vulgare L.) were tested in a factorial salinity-drought experiment, combining three levels of salinity and two levels of drought. The two drought treatments were obtained by applying irrigation water when the pre-dawn

  2. Zinc biofortification of cereals: rice differs from wheat and barley

    NARCIS (Netherlands)

    Stomph, T.J.; Jiang, W.; Struik, P.C.

    2009-01-01

    In their review, mainly focused on bread wheat (Triticum aestivum), durum wheat (Triticum durum) and barley (Hordeum vulgare), Palmgren et al. 1 M.G. Palmgren et al., Zinc biofortification of cereals: problems and solutions, Trends Plant Sci. 13 (2008), pp. 464–473. Article | PDF (905 K) | View Reco

  3. Isolating Barley (Hordeum vulgare L.) B1 Hordein Gene Promoter ...

    African Journals Online (AJOL)

    Yomi

    2012-04-10

    Apr 10, 2012 ... region of B1 hordein gene was isolated from the genomic DNA of Walfajre and Alger barley by ... plasmid DNA extraction kits were provided from Bioneer ... The E. coli competent cells were used for transformation by 5 µL of.

  4. Characterization of Gibberellin Receptor Mutants of Barley (Hordeum vulgare L.)

    Institute of Scientific and Technical Information of China (English)

    Peter M.Chandler; Carol A.Harding; Anthony R.Ashton; Mark D.Mulcair; Nicholas E.Dixon; Lewis N.Mander

    2008-01-01

    The sequence of Gidl (a gene for a gibberellin (GA) receptor from rice) was used to identify a putative orthoIogue from barley.This was expressed in E.coil,and produced a protein that was able to bind GA in vitro with both structural specificity and saturability.Its potential role in GA responses was investigated using barley mutants with reduced GA sensitivity (gsel mutants).Sixteen different gsel mutants each carried a unique nucleotide substitution in this sequence.In all but one case,these changes resulted in single amino acid substitutions,and,for the remaining mutant,a substitution in the 5' untranslated region of the mRNA is proposed to interfere with translation initiation.There was perfect linkage in segregating populations between new mutant alleles and the gsel phenotype,leading to the conclusion that the putative GID1 GA receptor sequence in barley corresponds to the Gsel locus.Determination of endogenous GA contents in one of the mutants revealed enhanced accumulation of bioactive GA1,and a deficit of C20 GA precursors.All of the gsel mutants had reduced sensitivity to exogenous GA3,and to AC94377 (a GA analogue) at concentrations that are normally 'saturating',but,at much higher concentrations,there was often a considerable response.The comparison between barley and rice mutants reveals interesting differences between these two cereal species in GA hormonal physiology.

  5. Evaluation of a malting barley quality assessment system

    NARCIS (Netherlands)

    Lonkhuijsen, H.J. van; Douma, A.C.; Angelino, S.A.G.F.

    1998-01-01

    New malting barley varieties are annually tested for their malting and brewing potential according to a field trial set-up combined with quality evaluation on pilot scale. To assess the effects of trial year and location on quality evaluation data, a data base consisting of quality data from Dutch

  6. Evaluation of a malting barley quality assessment system

    NARCIS (Netherlands)

    Lonkhuijsen, H.J. van; Douma, A.C.; Angelino, S.A.G.F.

    1998-01-01

    New malting barley varieties are annually tested for their malting and brewing potential according to a field trial set-up combined with quality evaluation on pilot scale. To assess the effects of trial year and location on quality evaluation data, a data base consisting of quality data from Dutch m

  7. Spatial aggregation of pathotypes of barley powdery mildew

    DEFF Research Database (Denmark)

    O'Hara, R.B.; Brown, J.K.M.

    1997-01-01

    Aggregation in the distribution of pathotypes of Erysiphe graminis f.sp. hordei, the barley powdery mildew pathogen, was investigated in field plots of 'Golden Promise', 'Proctor' and 'Tyra'. 'Golden Promise' and 'Proctor' have no effective mildew resistance alleles, whereas 'Tyra' has Mla1, which...

  8. Latent manganese deficiency increases transpiration in barley (Hordeum vulgare)

    DEFF Research Database (Denmark)

    Hebbern, Christopher Alan; Laursen, Kristian Holst; Ladegaard, Anne Hald

    2009-01-01

    To investigate if latent manganese (Mn) deficiency leads to increased transpiration, barley plants were grown for 10 weeks in hydroponics with daily additions of Mn in the low nM range. The Mn-starved plants did not exhibit visual leaf symptoms of Mn deficiency, but Chl a fluorescence measurements...

  9. Microgeographic Edaphic Differentiation in Hordein Polymorphisms of Wild Barley

    DEFF Research Database (Denmark)

    Nevo, E.; Beiles, A.; Storch, N.

    1983-01-01

    and topography. Likewise, significant correlations were found between hordein phenotypes and allozyme types detected in a previous study. Our results suggest that at least part of the hordein polymorphisms in wild barley is adaptive and selected by soil and topographic differences over very short distances....

  10. Wheat and barley seed systems in Ethiopia and Syria

    NARCIS (Netherlands)

    Bishaw, Z.

    2004-01-01

    Keywords: Wheat,Triticumspp., Barley,Hordeumvulgare L., Seed Systems, Formal Seed Sector, Informal Seed Sector, National Seed Program, Seed Source, Seed Selection, Seed Management, Seed Quality,

  11. Reclamation of Sodic-Saline Soils. Barley Crop Response

    Directory of Open Access Journals (Sweden)

    Giovanna Cucci

    2008-12-01

    Full Text Available The research was aimed at assessing the salinity and sodicity effects of two soil types submitted to correction on barley crop. The two soils, contained in cylindrical pots (0.40 m in size and 0.60 m h supplied with a bottom valve for the collection of drainage water and located under shed to prevent the leaching action of rainfall, were clay-textured and saline and sodic-saline at barley seeding, as they had been cultivated for 4 consecutive years with different herbaceous species irrigated with 9 types of brackish water. In 2002-2003 the 2 salinized and sodium-affected soils (ECe and ESP ranging respectively from 5.84-20.27 dSm-1 to 2.83-11.19%, submitted to correction, were cultivated with barley cv Micuccio, and irrigated with fresh water (ECw = 0.5 dS m-1 and SAR = 0.45 whenever 30% of the maximum soil available moisture was lost by evapotranspiration. Barley was shown to be a salt-tolerant species and did not experience any salt stress when grown in soils with an initial ECe up to 11 dS m-1. When it was grown in more saline soils (initial ECe of about 20 dS m-1, despite the correction, it showed a reduction in shoot biomass and kernel yield by 26% and 36% respectively, as compared to less saline soils.

  12. Wheat and barley seed systems in Ethiopia and Syria

    NARCIS (Netherlands)

    Bishaw, Z.

    2004-01-01

    Keywords: Wheat,Triticumspp., Barley,Hordeumvulgare L., Seed Systems, Formal Seed Sector, Informal Seed Sector, National Seed Program, Seed Source, Seed Selection, Seed Management, Seed Quality,

  13. Wheat and barley differently affect porcine intestinal microbiota

    DEFF Research Database (Denmark)

    Weiss, Eva; Aumiller, Tobias; Spindler, Hanns K

    2016-01-01

    Diet influences the porcine intestinal microbial ecosystem. Barrows were fitted with ileal T-cannulas to compare short-term effects of eight different wheat or barley genotypes and period-to-period effects on seven bacterial groups in ileal digesta and faeces by qPCR. Within genotypes of wheat an...

  14. Zinc biofortification of cereals: rice differs from wheat and barley

    NARCIS (Netherlands)

    Stomph, T.J.; Jiang, W.; Struik, P.C.

    2009-01-01

    In their review, mainly focused on bread wheat (Triticum aestivum), durum wheat (Triticum durum) and barley (Hordeum vulgare), Palmgren et al. 1 M.G. Palmgren et al., Zinc biofortification of cereals: problems and solutions, Trends Plant Sci. 13 (2008), pp. 464–473. Article | PDF (905 K) | View Reco

  15. Genetic diversity of some Saudi barley (Hordeum Vulgare L ...

    African Journals Online (AJOL)

    enoh

    2012-03-13

    Mar 13, 2012 ... These results could be used for barley germplasm management in terms of biodiversity ... animal feed, malt manufactures and human food. Its importance ... indigenous crop genetic resources of KSA are potentially threatened and ... and Hayes, 2002; Turpeinen et al., 2003; Nevo et al.,. 2005; Chaabane et ...

  16. Evaluation of Wheat and Barley Cultivars Tolerance to Metribuzine Application

    Directory of Open Access Journals (Sweden)

    E Izadi Darbandi

    2013-08-01

    Full Text Available In order to study of barely and wheat cultivars tolerance to metribuzin, a factorial experiment was conducted as a completely randomized design, with three replications in Greenhouse of Agricultural Research at Ferdowsi University of Mashhad. Treatments included wheat cultivars (Backcross roshan, Cross Arvand, Bahar, Sepahan, Gascosion, Sayonez, Bam garmsiry, Garmsiri, Ghods, Pishtaz, Chamran and Shoori 6, barely cultivars (Macouyi, Karoon and Bahman and metribuzin application rates ( 0, 175, 350, 700, 1050, 1400 and 2100 gr. ai.ha-1. Metribuzine was applied at 3-4 leaf stage and 3 weeks after herbicide spraying, plants survival and their biomass were determined. Results showed that metribuzin application had a significant effect (p≤0.01 on barley and wheat dry weight. Based on results, mertibuzin application did not affect on barley cultivars up to 30 g.a.i.ha-1 but in wheat varieties lead to significant reduction in their biomass and survival. Increasing of metribuzin rates reduced wheat and barley cultivars biomass (p≤0.01. Barely varieties were less sensitive than wheat cultivars to metribuzine. The highest and the lowest ED50 in wheat cultivars were observed in cross arvand (940 and shoori (25 varieties, respectively. In barley cultivars the highest and lowest ED50 were observed in Macouyi (614 and Karoon (396, respectively.

  17. Barley starch bioengineering for high phosphate and amylose

    DEFF Research Database (Denmark)

    Blennow, Per Gunnar Andreas; Carciofi, Massimiliano; Shaik, Shahnoor Sultana

    2011-01-01

    of the three genes encoding the starch-branching enzymes SBEI, SBEIIa, and SBEIIb using a triple RNAi chimeric hairpin construct we generated a virtually amylopectin-free barley. The grains of the transgenic lines were shrunken and had a yield of around 80% of the control line. The starch granules were...

  18. Giemsa C-banding of Barley Chromosomes. III

    DEFF Research Database (Denmark)

    Linde-Laursen, Ib

    1979-01-01

    Sixty-five homozygous barley lines, i.e. coming from chromosome-doubled monoploids derived from female gametes of F1 plants by the bulbosum method, segregated as expected in accordance with a 1:1-ratio for C-bands at two locations on chromosome 3 and at one location on chromosome 6. C-bands at one...

  19. The Mutation Frequency in Different Spike Categories in Barley

    DEFF Research Database (Denmark)

    Frydenberg, O.; Doll, Hans; Sandfær, J.

    1964-01-01

    After gamma irradiation of barley seeds, a comparison has been made between the chlorophyll-mutant frequencies in X1 spikes that had multicellular bud meristems in the seeds at the time of treatment (denoted as pre-formed spikes) and X1 spikes having no recognizable meristems at the time...

  20. Synthesis of the major storage protein, hordein, in barley

    DEFF Research Database (Denmark)

    Giese, Nanna Henriette; Andersen, B.; Doll, Hans

    1983-01-01

    A liquid culture system for culturing detached spikes of barley (Hordeum vulgare L.) at different nutritional levels was established. The synthesis of hordein polypeptides was studied by pulse-labeling with [14C]sucrose at different stages of development and nitrogen (N) nutrition. All polypeptid...

  1. Wheat and barley exposure to nanoceria: Implications for agricultural productivity

    Science.gov (United States)

    The impacts of man-made nanomaterials on agricultural productivity are not yet well understood. A soil microcosm study was performed to assess the physiological, phenological, and yield responses of wheat (Triticum aestivum) and barley (Hordeum vulgare L.) exposed to nanoceria (n...

  2. Enzyme superoxide dismutase in grain of barley and malt

    Directory of Open Access Journals (Sweden)

    Natálie Belcrediová

    2006-01-01

    Full Text Available The aim of the work was modification of superoxide dismutase enzyme (SOD, EC 1.15.1.1 activity analysis in barley grain and identical malts with using of the Ransod set. This set from company Randox were used for enzyme determination in blood samples. This method employs xanthine and xanthine oxidase to generate superoxide radicals, which react with tetrazolium chloride to form a red formazan dye. SOD is classified as natural antioxidants and enzyme plays a significant role at detoxication of products of molecular oxygen degradation. The largest rate of SOD occurs in embryo of barley grain. Its presence in barley grain and malt thus inhibits rancidity of grain during storage and undesirable beer flavour. The line Wabet x Washonubet (in grain-104,93 and malt 152,42 U/g dry matter and the variety Annabell (104,65 a 147,21 U/g dry matter had the highest activity of SOD in grain and malt of barley while the lowest activity was measured in the line KM 1910 (73,15 a 88,16 U/g dry matter and variety Tolar (74,34 a 96,44 U/g dry matter.

  3. Registration of Harriman low-phytate, hulled spring barley

    Science.gov (United States)

    The Agricultural Research Service, U.S. Department of Agriculture (USDA-ARS), has released 'Harriman', (Hordeum vulgare L.) (Reg. No. xxxxxx, P.I. xxxxxx). Harriman is a hulled, low-phytate barley, the second to be developed and released by the USDA-ARS. Compared to the previously released hulled, l...

  4. Biotic stress in barley: disease problems and solutions

    Science.gov (United States)

    Barley (Hordeum vulgare L.) is cultivated over a wider geographic range than almost any other major crop species. It can be found growing from the tropics to the high latitudes and from the seacoast to the highest arable mountaintops. On marginal lands where alkaline soils, drought, or cold summer t...

  5. Radiation hybrid map of barley chromosome 3H

    Science.gov (United States)

    Assembly of the barley genome is complicated by its large size (5.1 Gb) and proportion of repetitive elements (84%). This process is facilitated by high resolution maps for aligning BAC contigs along chromosomes. Available genetic maps; however, do not provide accurate information on the physical po...

  6. Aspects of the barley seed proteome during development and germination

    DEFF Research Database (Denmark)

    Finnie, Christine; Maeda, K.; Østergaard, O.;

    2004-01-01

    Analysis of the water-soluble barley seed proteome has led to the identification of proteins by MS in the major spots on two-dimensional gels covering the pi ranges 4-7 and 6-11. This provides the basis for in-depth studies of proteome changes during seed development and germination, tissue...

  7. Involvement of Alternative Splicing in Barley Seed Germination.

    Science.gov (United States)

    Zhang, Qisen; Zhang, Xiaoqi; Wang, Songbo; Tan, Cong; Zhou, Gaofeng; Li, Chengdao

    2016-01-01

    Seed germination activates many new biological processes including DNA, membrane and mitochondrial repairs and requires active protein synthesis and sufficient energy supply. Alternative splicing (AS) regulates many cellular processes including cell differentiation and environmental adaptations. However, limited information is available on the regulation of seed germination at post-transcriptional levels. We have conducted RNA-sequencing experiments to dissect AS events in barley seed germination. We identified between 552 and 669 common AS transcripts in germinating barley embryos from four barley varieties (Hordeum vulgare L. Bass, Baudin, Harrington and Stirling). Alternative 3' splicing (34%-45%), intron retention (32%-34%) and alternative 5' splicing (16%-21%) were three major AS events in germinating embryos. The AS transcripts were predominantly mapped onto ribosome, RNA transport machineries, spliceosome, plant hormone signal transduction, glycolysis, sugar and carbon metabolism pathways. Transcripts of these genes were also very abundant in the early stage of seed germination. Correlation analysis of gene expression showed that AS hormone responsive transcripts could also be co-expressed with genes responsible for protein biosynthesis and sugar metabolisms. Our RNA-sequencing data revealed that AS could play important roles in barley seed germination.

  8. Dormant barley aleurone shows heterogeneity and a specific cytodifferentiation

    NARCIS (Netherlands)

    Schuurink, R.C.; Bakhuizen, R.; Libbenga, K.R.; Boulanger, F.; Sinjorgo, K.M.C.

    1997-01-01

    In response to gibberellic acid, aleurone layers isolated from dormant barley (Hordeum distichum L. cv. Triumph) kernels produced significantly less alpha-amylase than aleurones from non-dormant kernels. Light microscopical investigations using the dye acridine orange as well as electron microscopic

  9. Expression of lipoxygenase isoenzymes in developing barley grains

    NARCIS (Netherlands)

    Schmitt, N.F.; Mechelen, J.R. van

    1997-01-01

    Expression of lipoxygenase was studied in whole developing barley grains from 5 days after flowering (DAF) to full maturity. Lipoxygenase showed two distinct peaks of activity. The first peak of activity occurred in the early stages of grain development from 5 until 20 DAF, whereas the second peak o

  10. Comparison of lignin peroxidase and horseradish peroxidase for catalyzing the removal of nonylphenol from water.

    Science.gov (United States)

    Dong, Shipeng; Mao, Liang; Luo, Siqiang; Zhou, Lei; Feng, Yiping; Gao, Shixiang

    2014-02-01

    Concentrations of aqueous-phase nonylphenol (NP), a well-known endocrine-disrupting chemical, are shown to be reduced effectively via reaction with lignin peroxidase (LiP) or horseradish peroxidase (HRP) and hydrogen peroxide. We systematically assessed their reaction efficiencies at varying conditions, and the results have confirmed that the catalytic performance of LiP toward NP was more efficient than that of HRP under experimental conditions. Mass spectrum analysis demonstrated that polymerization through radical-radical coupling mechanism was the pathway leading to NP transformation. Our molecular modeling with the assistance of ab initio suggested the coupling of NP likely proceeded via covalent bonding between two NP radicals at their unsubstituted carbons in phenolic rings. Data from acute immobilization tests with Daphnia confirm that NP toxicity is effectively eliminated by LiP/HRP-catalyzed NP removal. The findings in this study provide useful information for understanding LiP/HRP-mediated NP reactions, and comparison of enzymatic performance can present their advantages for up-scale applications in water/wastewater treatment.

  11. Lignin degradation by a white-rot fungus lacking lignin peroxidase and manganese peroxidase

    Energy Technology Data Exchange (ETDEWEB)

    Eggert, C.B.; Eriksson, K.E.L. [Univ. of Georgia, Athens, GA (United States)

    1996-10-01

    Phanerochaete chrysosporium has been the organism of choice for studies of lignin degradation and much of this work has focused on two phenol oxidases, lignin peroxidase (LiP) and manganese peroxidase (MnP), secreted by the fungus under ligninolytic conditions. However, many white-rot fungi, including a number of aggressive lignin degraders, seem to operate without expressing LiP activity. Laccase is another phenol oxidase that white-rot fungi often produce. However, the role played by laccase in lignin degradation has remained obscured since its low redox potential appeared to make it incapable of oxidizing non-phenolic lignin constituents. We have identified, Pychnoporus cinnabarinus lacking both LiP and MnP, but a high producer of laccase, to degrade lignin as efficiently as UP producing fungi. We have found that P. cinnabarinus, to overcome the redox potential barrier for laccase, produces a mediator for oxidation of non-phenolic lignin structures. This is the first description of how laccase may be used in a biological system for the degradation of lignin.

  12. Oxidation of wheat straw lignin by fungal lignin peroxidase, manganese peroxidase and laccase: A comparative study

    Energy Technology Data Exchange (ETDEWEB)

    Martinez-Ingo, M.J.; Kurek, B. [Laboratorie de Chimie Biologique, Thiverval-Grignon (France)

    1996-10-01

    Lignin peroxidase (LiP), manganese peroxidase (MnP) from Phanerochaete chrysosporium and laccase from Pleurotus eryngii were separately used to degrade alkali wheat straw lignin (AL). In order to characterize the catalytic action of the different enzymes, the chemical structure and the hydrodynamic properties of the treated lignin were analyzed by thioacidolysis-gas chromatography and molecular size exclusion chromatography. The results confirmed that only LiP was able to degrade guiacyl (G) and syringyl (S) structures in non-phenolic methylated lignins. However, provided that some phenolic terminal structures are present, MnP and laccase were able to degrade the non-phenolic portion of the polymer linked by {beta}-O-4 alkyl aryl ether bonds. This suggested that the oxidative reactions catalyzed in alkali straw lignin could progress through bond cleavages generating phenoxy radicals. The molecular size distribution of both thioacidolysis products and the oxidized polymer showed that AL underwent condensation side-reactions regardless of the enzyme treatment, but only LiP oxidation led to the increase in the hydrodynamic volume of the recovered lignin. This indicated that modification of enzymes by bonding patterns in lignin is not always associated with alterations in the spatial network of the polymer.

  13. Roles of horseradish peroxidase in response to terbium stress.

    Science.gov (United States)

    Zhang, Xuanbo; Wang, Lihong; Zhou, Qing

    2014-10-01

    The pollution of the environment by rare earth elements (REEs) causes deleterious effects on plants. Peroxidase plays important roles in plant response to various environmental stresses. Here, to further understand the overall roles of peroxidase in response to REE stress, the effects of the REE terbium ion (Tb(3+)) on the peroxidase activity and H2O2 and lignin contents in the leaves and roots of horseradish during different growth stages were simultaneously investigated. The results showed that after 24 and 48 h of Tb(3+) treatment, the peroxidase activity in horseradish leaves decreased, while the H2O2 and lignin contents increased. After a long-term (8 and 16 days) treatment with Tb(3+), these effects were also observed in the roots. The analysis of the changes in peroxidase activity and H2O2 and lignin contents revealed that peroxidase plays important roles in not only reactive oxygen species scavenging but also cell wall lignification in horseradish under Tb(3+) stress. These roles were closely related to the dose of Tb(3+), duration of stress, and growth stages of horseradish.

  14. The impact of thiol peroxidases on redox regulation.

    Science.gov (United States)

    Flohé, Leopold

    2016-01-01

    The biology of glutathione peroxidases and peroxiredoxins is reviewed with emphasis on their role in metabolic regulation. Apart from their obvious function in balancing oxidative challenge, these thiol peroxidases are not only implicated in orchestrating the adaptive response to oxidative stress, but also in regulating signaling triggered by hormones, growth factors and cytokines. The mechanisms presently discussed comprise dampening of redox-sensitive regulatory processes by elimination of hydroperoxides, suppression of lipoxygenase activity, committing suicide to save H2O2 for signaling, direct binding to receptors or regulatory proteins in a peroxidase activity-independent manner, or acting as sensors for hydroperoxides and as transducers of oxidant signals. The various mechanistic proposals are discussed in the light of kinetic data, which unfortunately are scarce. Taking into account pivotal criteria of a meaningful regulatory circuit, kinetic plausibility and specificity, the mechanistic concepts implying a direct sensor/transducer function of the thiol peroxidases appear most appealing. With rate constants for the reaction with hydroperoxide of 10(5)-10(8) M(-1) s(-1), thiol peroxidases are qualified as kinetically preferred hydroperoxide sensors, and the ability of the oxidized enzymes to react with defined protein thiols lends specificity to the transduction process. The versatility of thiol peroxidases, however, allows multiple ways of interaction with regulatory pathways.

  15. Engineering a fungal peroxidase that degrades lignin at very acidic pH

    NARCIS (Netherlands)

    Fernandez-Fueyo, E.; Ruiz-Duenas, F.J.; Martinez, A.T.

    2014-01-01

    Background Ligninolytic peroxidases are divided into three families: manganese peroxidases (MnPs), lignin peroxidases (LiPs), and versatile peroxidases (VPs). The latter two are able to degrade intact lignins, as shown using nonphenolic lignin model compounds, with VP oxidizing the widest range of r

  16. Engineering a fungal peroxidase that degrades lignin at very acidic pH

    NARCIS (Netherlands)

    Fernandez-Fueyo, E.; Ruiz-Duenas, F.J.; Martinez, A.T.

    2014-01-01

    Background Ligninolytic peroxidases are divided into three families: manganese peroxidases (MnPs), lignin peroxidases (LiPs), and versatile peroxidases (VPs). The latter two are able to degrade intact lignins, as shown using nonphenolic lignin model compounds, with VP oxidizing the widest range of

  17. Optimization of lignin peroxidase, manganese peroxidase, and Lac production from Ganoderma lucidum under solid state fermentation of pineapple leaf

    OpenAIRE

    Sudha Hariharan; Padma Nambisan

    2013-01-01

    This study was undertaken to isolate ligninase-producing white-rot fungi for use in the extraction of fibre from pineapple leaf agriwaste. Fifteen fungal strains were isolated from dead tree trunks and leaf litter. Ligninolytic enzymes (lignin peroxidase (LiP), manganese peroxidase (MnP), and laccase (Lac)), were produced by solid-state fermentation (SSF) using pineapple leaves as the substrate. Of the isolated strains, the one showing maximum production of ligninolytic enzymes was identified...

  18. iTAG Barley: A 9-12 curriculum to explore inheritance of traits and genes using Oregon Wolfe barley

    Science.gov (United States)

    Segregating plants from the Informative & Spectacular Subset (ISS) of the Oregon Wolfe doubled haploid barley (OWB) population are easily grown on a lighted window bench in the classroom. These lines originate from a wide cross and have exceptionally diverse and dramatic phenotypes, making this an i...

  19. Tocopherols and tocotrienols in barley oil prepared from germ and other fractions from scarification and sieving of hulless barley

    Science.gov (United States)

    Two cultivars of hulless barley (Doyce and Merlin), were scarified to abrade the outer layers of the kernels (germ, pericarp, and aleurone). The resulting scarification fines fractions were then separated into four particle size subfractions using sieves. Each of the size subfractions was then extr...

  20. Barley HvPAPhy_a as transgene provides high and stable phytase activities in mature barley straw and in grains.

    Science.gov (United States)

    Holme, Inger Baeksted; Dionisio, Giuseppe; Madsen, Claus Krogh; Brinch-Pedersen, Henrik

    2017-04-01

    The phytase purple acid phosphatase (HvPAPhy_a) expressed during barley seed development was evaluated as transgene for overexpression in barley. The phytase was expressed constitutively driven by the cauliflower mosaic virus 35S-promoter, and the phytase activity was measured in the mature grains, the green leaves and in the dry mature vegetative plant parts left after harvest of the grains. The T2 -generation of HvPAPhy_a transformed barley showed phytase activity increases up to 19-fold (29 000 phytase units (FTU) per kg in mature grains). Moreover, also in green leaves and mature dry straw, phytase activities were increased significantly by 110-fold (52 000 FTU/kg) and 57-fold (51 000 FTU/kg), respectively. The HvPAPhy_a-transformed barley plants with high phytase activities possess triple potential utilities for the improvement of phosphate bioavailability. First of all, the utilization of the mature grains as feed to increase the release of bio-available phosphate and minerals bound to the phytate of the grains; secondly, the utilization of the powdered straw either directly or phytase extracted hereof as a supplement to high phytate feed or food; and finally, the use of the stubble to be ploughed into the soil for mobilizing phytate-bound phosphate for plant growth. © 2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

  1. Allelopathic effects of barley straw on germination and seedling growth of corn, sugar beet and sunflower

    Directory of Open Access Journals (Sweden)

    mohamad taghi naseri poor yazdi

    2009-06-01

    Full Text Available Allelopathic effects of barley straw and root on germination and growth of maize, sugar beet, and sunflower were investigated under glasshouse and laboratory experiments in Faculty of Agriculture, Ferdowsi University of Mashhad in 2006. The glasshouse experiment was designed based on randomized complete block design with three replications, treatments included: 0, 200, 400, 600 g/m² of grounded barley straw and also 0 and 50 g/m2 barley root. A laboratory experiment was carried out in order to study the effect of different concentrations of barley water extracts on germination and seedling characteristics of corn, sugar beet and sunflower. Treatments in laboratory trial included 0, 33, 50 and 100 percent of barley extracts. Results showed that leaf area of corn was significantly affected by barley straw treatments. Shoot dry matter and seed weight per plant in corn , leaf and tuber weight in sugar beet and leaf , stem weights , plant per plant in corn , leaf and tuber weight in sugar beet and leaf, stem weights, plant height, head diameter, head weight and seed weight in sunflower were significantly higher in treatment of 50g/m² barley roots. Crop seed germination decreased with increasing the amount of barley straw. The best germination response to barley extract was observed in corn. Maize radicle weight was significantly decreased with increasing concentration of barley water extract.

  2. NMR studies of recombinant Coprinus peroxidase and three site-directed mutants. Implications for peroxidase substrate binding.

    Science.gov (United States)

    Veitch, N C; Tams, J W; Vind, J; Dalbøge, H; Welinder, K G

    1994-06-15

    Proton nuclear magnetic resonance spectroscopy has been used to characterise and compare wild-type fungal and recombinant Coprinus cinereus peroxidase (CIP) and three mutants in which Gly156 and/or Asn157 was replaced by Phe. Analysis of one- and two-dimensional NMR spectra of recombinant CIP was undertaken for comparison with the fungal enzyme and in order to establish a meaningful basis for solution studies of CIP mutants. Proton resonance assignments of haem and haem-linked residues obtained for the cyanide-ligated form of recombinant CIP revealed a high degree of spectral similarity with those of lignin and manganese-dependent peroxidases and extend previously reported NMR data for fungal CIP. The three mutants examined by NMR spectroscopy comprised site-specific substitutions made to a region of the structure believed to form part of the peroxidase haem group access channel for substrate and ligand molecules. Proton resonances of the aromatic side-chains of Phe156 and Phe157 were found to have similar spectral characteristics to those of two phenylalanine residues known to be involved in the binding of aromatic donor molecules to the plant peroxidase, horseradish peroxidase isoenzyme C. The results are discussed in the context of complementary reactivity studies on the mutants in order to develop a more detailed understanding of aromatic donor molecule binding to fungal and plant peroxidases.

  3. Role of peroxidases in the compensation of cytosolic ascorbate peroxidase knockdown in rice plants under abiotic stress.

    Science.gov (United States)

    Bonifacio, Aurenivia; Martins, Marcio O; Ribeiro, Carolina W; Fontenele, Adilton V; Carvalho, Fabricio E L; Margis-Pinheiro, Márcia; Silveira, Joaquim A G

    2011-10-01

    Current studies, particularly in Arabidopsis, have demonstrated that mutants deficient in cytosolic ascorbate peroxidases (APXs) are susceptible to the oxidative damage induced by abiotic stress. In contrast, we demonstrate here that rice mutants double silenced for cytosolic APXs (APx1/2s) up-regulated other peroxidases, making the mutants able to cope with abiotic stress, such as salt, heat, high light and methyl viologen, similar to non-transformed (NT) plants. The APx1/2s mutants exhibited an altered redox homeostasis, as indicated by increased levels of H₂O₂ and ascorbate and glutathione redox states. Both mutant and NT plants exhibited similar photosynthesis (CO₂) assimilation and photochemical efficiency) under both normal and stress conditions. Overall, the antioxidative compensatory mechanism displayed by the mutants was associated with increased expression of OsGpx genes, which resulted in higher glutathione peroxidase (GPX) activity in the cytosolic and chloroplastic fractions. The transcript levels of OsCatA and OsCatB and the activities of catalase (CAT) and guaiacol peroxidase (GPOD; type III peroxidases) were also up-regulated. None of the six studied isoforms of OsApx were up-regulated under normal growth conditions. Therefore, the deficiency in cytosolic APXs was effectively compensated for by up-regulation of other peroxidases. We propose that signalling mechanisms triggered in rice mutants could be distinct from those proposed for Arabidopsis.

  4. Malate dehydrogenase isozymes (MDH; EC 1.1.1.37) in long-term callus culture of Cereus peruvianus (Cactaceae) exposed to sugar and temperature stress.

    Science.gov (United States)

    Jorge, I C; Mangolin, C A; Machado, M F

    1997-06-01

    Malate dehydrogenase (MDH; EC 1.1.1.37) isozymes in long-term callus tissue culture of Cereus peruvianus were studied in starch gel electrophoresis to investigate the control of differential Mdh gene expression under sugar and temperature stress. While two cytosol MDH isozymes showed an unchanged phenotype when the callus tissues were transferred to medium maintained at 22 or 37 degrees C and containing different concentrations of sucrose, glucose, and fructose, the different combinations of five mitochondrial MDH (mtMDH) and two micro-body MDH (mbMDH) showed different MDH isozyme patterns in the callus populations. Differential expression of mtMDH isozymes seems to be modulated at the posttranslational level in callus tissues exposed to different concentrations and types of sugar and to high-temperature and low-temperature stress. An inductor effect on the expression of mbMDH isozymes was observed under stress conditions and in long-term callus tissue, and they may also present different responses.

  5. Barley HvPAPhy_a as transgene provides high and stable phytase activities in mature barley straw and in grains

    DEFF Research Database (Denmark)

    Holme, Inger; Dionisio, Giuseppe; Madsen, Claus Krogh

    2017-01-01

    The phytase purple acid phosphatase (HvPAPhy_a) expressed during barley seed development was evaluated as transgene for overexpression in barley. The phytase was expressed constitutively driven by the cauliflower mosaic virus 35S-promoter, and the phytase activity was measured in the mature grain...

  6. Effect of pH and Recombinant Barley (Hordeum vulgare L.) Endoprotease B2 on Degradation of Proteins in Soaked Barley

    DEFF Research Database (Denmark)

    Christensen, Jesper Bjerg; Dionisio, Giuseppe; Poulsen, Hanne Damgaard

    2014-01-01

    Nonfermented soaking of barley feedstuff has been established as an in vitro procedure prior to the feeding of pigs as it can increase protein digestibility. In the current study, two feed cultivars of barley (Finlissa and Zephyr) were soaked in vitro either nonbuffered or buffered at pH 3.6 and ...

  7. Peroxisomal membrane manganese superoxide dismutase: characterization of the isozyme from watermelon (Citrullus lanatus Schrad.) cotyledons.

    Science.gov (United States)

    Rodríguez-Serrano, María; Romero-Puertas, María C; Pastori, Gabriela M; Corpas, Francisco J; Sandalio, Luisa M; del Río, Luis A; Palma, José M

    2007-01-01

    In this work the manganese superoxide dismutase (Mn-SOD) bound to peroxisomal membranes of watermelon cotyledons (Citrullus lanatus Schrad.) was purified to homogeneity and some of its molecular properties were determined. The stepwise purification procedure consisted of ammonium sulphate fractionation, batch anion-exchange chromatography, and anion-exchange and gel-filtration column chromatography using a fast protein liquid chromatography system. Peroxisomal membrane Mn-SOD (perMn-SOD; EC 1.15.1.1) was purified 5600-fold with a yield of 2.6 mug of enzyme g(-1) of cotyledons, and had a specific activity of 480 U mg(-1) of protein. The native molecular mass determined for perMn-SOD was 108 000 Da, and it was composed of four equal subunits of 27 kDa, which indicates that perMn-SOD is a homotetramer. Ultraviolet and visible absorption spectra of the enzyme showed a shoulder at 275 nm and two absorption maxima at 448 nm and 555 nm, respectively. By isoelectric focusing, a pI of 5.75 was determined for perMn-SOD. In immunoblot assays, purified perMn-SOD was recognized by a polyclonal antibody against Mn-SOD from pea leaves, and the peroxisomal enzyme rapidly dissociated in the presence of dithiothreitol and SDS. The potential binding of the Mn-SOD isozyme to the peroxisomal membrane was confirmed by immunoelectron microscopy analysis. The properties of perMn-SOD and the mitMn-SOD are compared and the possible function in peroxisomal membranes of the peripheral protein Mn-SOD is discussed.

  8. Dietary fish oil blocks carcinogen-induced down-regulation of colonic protein kinase C isozymes.

    Science.gov (United States)

    Jiang, Y H; Lupton, J R; Chapkin, R S

    1997-02-01

    In order to elucidate the influence of dietary constituents on colonic intracellular signal transduction, the effect of different fats on rat colonic epithelial protein kinase C (PKC) alpha (classical), delta (novel) and lambda-zeta (atypical) expression was determined in carcinogen-treated animals. Sprague-Dawley rats were provided with one of two fats (corn oil and fish oil); plus or minus the carcinogen azoxymethane (AOM) and killed at two time points (15 and 37 weeks) in a 2x2x2 factorial design. At 5 and 6 weeks of age, animals were injected s.c. with either AOM at a dose of 15 mg/kg body weight or saline once a week for 2 weeks and continued on the same diet until termination of the study. At 15 and 37 weeks after the second injection, 10 rats from each treatment group were killed. Colonic PKC alpha, delta and lambda-zeta steady-state protein and mRNA levels were determined using immunoblotting and relative quantitative polymerase chain reaction, respectively. Colonic mucosa from rats injected with AOM had significantly suppressed membrane and cytosolic PKC alpha and cytosolic lambda-zeta protein levels (P fish oil diets had significantly higher (P protein levels relative to animals fed corn oil diets. However, the effect of diet and AOM on the steady-state expression of PKC alpha, delta and zeta mRNA was not consistent with changes in the respective isozyme protein levels, suggesting regulation at the post-transcriptional level. These data demonstrate that dietary fish oil blocks the carcinogen-induced decrease in the steady-state levels of colonic mucosal PKC delta and lambda-zeta, which may in part explain why this fat source protects against colon cancer development.

  9. Biomonitoring of ecosystem degradation caused by CPO waste of Mentaya River in Central Kalimantan use of esterase isozyme electromorph method

    Directory of Open Access Journals (Sweden)

    PRABANG SETYONO

    2008-07-01

    Full Text Available The impact of CPO (Crude Palm Oil dock activity in Mentaya River of Central Borneo caused degradation of ecosystem, particularly on both mangrove and macrozoobenthos community. One of methods used for monitoring of ecosystem degradation was to determine species that were still survive under the polluted conditions. These survival species were assumed to synthesize alloenzyme that can be used as indicator. Alloenzyme was synthesized as an effort of adaptation processes toward environmental pressures caused by CPO spill on Mentaya River. Alloenzyme would be expressed as phenotypic and genotypic adaptation processes or phenotypic plasticity. Research was carried out, consisted of field research included collecting sample and environmental data (oil content, temperature, pH, electric conductivity and redox potential, and laboratory research included series analysis of water quality (DO, BOD, COD, pH, TSS, TDS and also alloenzyme content of Soneratia caseolaris L. and Macrobrachium rosenbergii de Man. The alloenzyme of root and leaves mangrove and prawn’s hepatopancreas was analyzed using Spencer starch gel electrophoresis modified method of exposed on sucrose solution. Separated components of alloenzyme were detected by special staining for Esterase isozyme. The results revealed that Soneratia caseolaris L. and Macrobrachium rosenbergii de Man were bioindicator organisms for the polluted site by oil spills from CPO loading activities. The polluted river water by oil spill from CPO activities decreased redox potential, DO, increased oil content, DHL, water temperature, pH sediment, pH water, TDS, BOD, COD, TSS. Gel electrophoretical analysis demonstrated that Mangrove Soneratia caseolaris synthesized alloenzyme consisted of complex enzymes such as EST in its root and leave cells. Those enzymes were nearly similar to those of Macrobrachium rosenbergii. The oil spill from CPO have ester bonding so its adaptation mechanism with release Esterase

  10. Diversity of esterase isozyme in Aegilops tauschii Cosson%节节麦的酯酶同工酶分析

    Institute of Scientific and Technical Information of China (English)

    兰秀锦; 刘登才; 魏育明; 颜泽红; 郑有良

    2001-01-01

    The esterase isozyme of 30 accessions of Aegilops tauschii were studied by means of polyarylamide gel electrophoresis. The results showed significant difference of esterase in all of the four stages i.e. seeding,Shooting, flag leaf and young ear, which patterns can be divided into 15 types. Ten accessions from middle reaches of the Yellow River belonged to two patterns with a similarity coefficient 0. 984. Two accessions of Xinjiang belonged to one pattern and was different from that of middle reaches of Yellow River. No same esterase isozyme has been found in the four stages. It showed that esterase isozyme related to growing and development of plants.%对30份不同来源的节节麦进行4个时期的酯酶同工酶分析。结果表明:不同来源节节麦的酯酶同工酶存在较大差异,共分成15种基本类型。我国黄河流域的10份节节麦被划分为2个基本类型,但二者关系极为相近;新疆节节麦与之有一定差异,但在相似系数≤0.820时可视为一类。所有材料在4个时期之间没有出现一个完全相同的酶带类型,说明酯酶同工酶随发育时期而不断变化。

  11. Determination of ergosterol levels in barley and malt varieties in the Czech Republic via HPLC.

    Science.gov (United States)

    Jedlicková, Lenka; Gadas, David; Havlová, Pavla; Havel, Josef

    2008-06-11

    Ergosterol is considered to be a suitable indicator of mold infestation in barley and malt. In this study ergosterol levels in different varieties of barley and malt produced in the Czech Republic were determined. A modified high-performance liquid chromatography (HPLC) method was statistically processed, validated (Effivalidation program), and applied to 124 samples of barley and malt. Ergosterol was isolated by extraction and saponification, and the quantification was performed using HPLC with diode array detection. The content of ergosterol ranged between the limit of detection (LOD) and 36.3 mg/kg in barley and between the LOD and 131.1 mg/kg in malt. Ergosterol is presumably connected with metabolites generated when barley grain is attacked by pathogens, and such barley often shows a high overfoaming (gushing) value. However, it was found that the content of ergosterol does not correlate with the degree of beer gushing.

  12. Peroxidase activation of cytoglobin by anionic phospholipids: Mechanisms and consequences.

    Science.gov (United States)

    Tejero, Jesús; Kapralov, Alexandr A; Baumgartner, Matthew P; Sparacino-Watkins, Courtney E; Anthonymutu, Tamil S; Vlasova, Irina I; Camacho, Carlos J; Gladwin, Mark T; Bayir, Hülya; Kagan, Valerian E

    2016-05-01

    Cytoglobin (Cygb) is a hexa-coordinated hemoprotein with yet to be defined physiological functions. The iron coordination and spin state of the Cygb heme group are sensitive to oxidation of two cysteine residues (Cys38/Cys83) and/or the binding of free fatty acids. However, the roles of redox vs lipid regulators of Cygb's structural rearrangements in the context of the protein peroxidase competence are not known. Searching for physiologically relevant lipid regulators of Cygb, here we report that anionic phospholipids, particularly phosphatidylinositolphosphates, affect structural organization of the protein and modulate its iron state and peroxidase activity both conjointly and/or independently of cysteine oxidation. Thus, different anionic lipids can operate in cysteine-dependent and cysteine-independent ways as inducers of the peroxidase activity. We establish that Cygb's peroxidase activity can be utilized for the catalysis of peroxidation of anionic phospholipids (including phosphatidylinositolphosphates) yielding mono-oxygenated molecular species. Combined with the computational simulations we propose a bipartite lipid binding model that rationalizes the modes of interactions with phospholipids, the effects on structural re-arrangements and the peroxidase activity of the hemoprotein.

  13. Peroxidase extraction from jicama skin peels for phenol removal

    Science.gov (United States)

    Chiong, T.; Lau, S. Y.; Khor, E. H.; Danquah, M. K.

    2016-06-01

    Phenol and its derivatives exist in various types of industrial effluents, and are known to be harmful to aquatic lives even at low concentrations. Conventional treatment technologies for phenol removal are challenged with long retention time, high energy consumption and process cost. Enzymatic treatment has emerged as an alternative technology for phenol removal from wastewater. These enzymes interact with aromatic compounds including phenols in the presence of hydrogen peroxide, forming free radicals which polymerize spontaneously to produce insoluble phenolic polymers. This work aims to extract peroxidase from agricultural wastes materials and establish its application for phenol removal. Peroxidase was extracted from jicama skin peels under varying extraction conditions of pH, sample-to-buffer ratio (w/v %) and temperature. Experimental results showed that extraction process conducted at pH 10, 40% w/v and 25oC demonstrated a peroxidase activity of 0.79 U/mL. Elevated temperatures slightly enhanced the peroxidase activities. Jicama peroxidase extracted at optimum extraction conditions demonstrated a phenol removal efficiency of 87.5% at pH 7. Phenol removal efficiency was ∼ 97% in the range of 30 - 40oC, and H2O2 dosage has to be kept below 100 mM for maximum removal under phenol concentration tested.

  14. BIOCHEMICAL GENETIC STUDIES ON CUTYLEFISH SEPIELLA MAINDRONI (CEPHALOPODA: SEPIIDAE)——ACTIVE LOCI SCREENING OF ISOZYME

    Institute of Scientific and Technical Information of China (English)

    郑小东; YutakaNatsukari; 王如才; 王昭萍; 李云

    2001-01-01

    Screening of 46 putative enzyme-coding loci and 4 different kinds of tissues of Sepiella maindroni de Rochebrone, 1884 for enzymatic activities using starch gel electrophoretic technique proved that the 21 enzymes such as AAT, AK, ALP, AP, CK, DIA, ES, FBP, G3PDH, GPI, GRS,IDH, LDH, MDH, MEP, MPI, NP, PGDH, PGM, SOD and XO* , were active to Sepiella maindroni after being stained. The tissue exhibiting stable and clear bands was also determined. Among tissues tested, mantle muscle tissue was the best for electrophoretic survey of isozymes. Buccal bulb muscle, eye and liver were fairly good for some special enzymes, such as DIA, ES, MPI, NT, etc.

  15. Protective effect of aqueous extract of Phyllanthus fraternus against bromobenzene induced changes on cytosolic glutathione S-transferase isozymes in rat liver

    Directory of Open Access Journals (Sweden)

    Sriram Gopi

    2017-07-01

    Full Text Available The aim of this study was to investigate beneficial effect of aqueous extract of Phyllanthus fraternus (AEPF on bromobenzene (BB induced changes on cytosolic glutathione S-transferase (GST isozymes in rat liver. Administration of BB significantly decreased the activity of GST, however, prior administration of AEPF prevented the BB induced decrease in GST activity. Further the cytosolic GSTs were purified from 3 groups of animals (control, BB and AEPF+BB administered and resolved into three protein bands on SDS-PAGE. Densitometric analysis showed a significant decrease in BB group compared to control. Further, 2D PAGE analysis resolved these proteins into 8 bands which were identified as five isozymes of alpha, two of Mu and one of theta by MALDI-TOF MS and also observed decreased levels of isozymes in BB group. However, on prior administration of AEPF significantly prevented the BB induced decrease in GSTs and restored to normal levels.

  16. Differential fitness of allelic isozymes in the marine gastropods Littorina punctata and Littorina neritoides, exposed to the environmental stress of the combined effects of cadmium and mercury pollution

    Science.gov (United States)

    Lavie, Batia; Nevo, Eviatar

    1987-07-01

    The present study tested the separate and the interactive pollution effects of cadmium and mercury on the electrophoretically detected allelic isozyme frequencies of the enzyme phosphoglucose isomerase for two species of littoral marine gastropods — Littorina punctata and L. neritoides — and the enzyme amino peptidase for L. neritoides. Our results indicate differential survivorship of allelic isozyme genotypes specific for each type of pollutant and for their interaction, as well as trends common to all pollutants. Theoretically the results reflect the adaptive nature of at least some allozymic genotypes in these marine gastropods and seem inconsistent with the neutral theory of allozyme polymorphisms. Practically, the results reinforce earlier conclusions that changes in the frequency of allelic isozymes may be used as a genetic monitor of pollution.

  17. Phylogeny and ontogeny of the phosphoglycerate mutases - III. Inactivation of rabbit muscle phosphoglycerate mutase (type M isozyme) by the sulfhydryl group reagents.

    Science.gov (United States)

    Carreras, J; Bosch, J; Mezquita, J

    1982-01-01

    1. The three phosphoglycerate mutase isozymes from mammals (types M, B and MB isozymes) differ in their sensitivity to the - SH group reagents. 2. Rabbit muscle phosphoglycerate mutase (type M isozyme) is reversibly inactivated by tetrathionate, rho-chloromercuribenzoate and Hg2+. 3. Titration with rho-chloromercuribenzoate shows the existence of two sulfhydryl groups per enzyme subunit, the modification of which produces a progressive decline in enzyme activity. 4. The apparent Km values for substrate and cofactor are not affected by tetrathionate treatment. 5. Phosphoglycerate mutase inactivated by tetrathionate and by rho-chloromercuribenzoate is unable to form the functionally active phosphorylenzyme when mixed with glycerate-2,3-P2, and is not protected by the cofactor against heating. 6. Glycerate-2,3-P2 protects against tetrathionate treatment, but fails to protect against Hg2+ and rho-chloromercuribenzoate inactivation.

  18. Retention and growth performance of chicks given low-phytate conventional or hull-less barleys

    Science.gov (United States)

    Four low-phytate, hulled lines, M2 422 (now referred to as barley lpa1-1), M2 635 (now referred to as barley lpa3-1), M2 955 and M2 1070 (now referred to as barley lpa2-1), and a "hulless" version of M2 422, were evaluated in a chick feeding experiment. The diets were provided in meal form, with the...

  19. Cultivar and Environmental Variation of β-glucan Content in Chinese Barleys

    Institute of Scientific and Technical Information of China (English)

    CHEN Jin-xin; Zhang Guo-ping; QIANG Xiao-lin; WANG Jun-mei; DING Shou-ren

    2002-01-01

    β-glucan is a polysaccharide compound closely related to the quality of barley used as malting,feed and food. Low β-glucan content is expected for brewing and feed barley, while high β-glucan content is desirable for food barley. The β-glucan content of barley genotypes collected from various areas of China as well as from Canada and Australia were assayed. Meanwhile a multi-locations trial was conducted to determineβ-glucan content of 10 barley cultivars in 8 locations for two successive planting years. The results showed that barley genotypes from Tibet and Xinjiang had higher β-glucan content and the genotypes with higher than 8%of β-glucan content were detected in Tibet barleys, being valuable for use in the development of healthy food.Barley cultivars being planted now in winter-sowing areas of China had basically the same β-glucan content as those from Canada and Australia. Barley seeds produced in Hangzhou had lower β-glucan content than seeds from the original areas. There was a highly significant difference in β-glucan content among 10 barleys, 8locations and between years. On an average of two years, Xiumei 3 and Kongpei 1 had the highest and lowestβ-glucan content, respectively, and Taian and Hangzhou produced the highest and lowest β-glucan content barley seeds, respectively. Analysis of AMMI model showed that interaction effect between cultivar and environment was highly significant in both experimental years, and was dependent on cuitivar, suggesting that it is important to plant the suitable cultivars in a particular area in order to obtain barley seeds with reasonableβ-glucan content.

  20. Application of Dual Coagulant (Alum + Barley in Removing Colour from Leachate

    Directory of Open Access Journals (Sweden)

    Shaylinda Mohd Zin Nur

    2017-01-01

    Full Text Available Coagulation/flocculation is one of the treatment method for highly polluted leachate. One of the main affecting factor for this process is the coagulant used. Coagulant is divided into natural and chemical coagulant. In the current study, Alum (chemical coagulant and barley (natural coagulant were used as dual coagulant. The aim of this study is to examine the effectiveness of dual coagulant made from alum and barley in removing colour from the effluent of Simpang Renggam landfill leachate aeration lagoon through coagulation/flocculation method. Coagulation/flocculation process with single alum coagulant, single barley coagulant and dual coagulant (alum+barley were examined by evaluating the optimum values of pH and dose. Optimum dose and pH for alum and barley as single coagulant were; 3 g/L & pH 5; 0.8 g/L & pH 6. Higher removal of colour was recorded for alum compared to barley. Application of alum and barley as dual coagulant had higher colour removal than alum and barley as single coagulant. The optimum pH and dose for dual coagulant were at pH 6, 3.0 g/L of alum and 0.8 g/L of barley respectively. However, at pH 6, 2 g/L alum and 1.6 g/L barley, the removal of colour was similar to alum at 3 g/L. It can be concluded that barley as coagulant aid able to reduce 33 % usage of alum at par removals of colour. Thus, the dual coagulant consist of alum and barley has the potential to be applied as a coagulant for leachate treatment.

  1. Barley Sprouts Extract Attenuates Alcoholic Fatty Liver Injury in Mice by Reducing Inflammatory Response

    Directory of Open Access Journals (Sweden)

    Yun-Hee Lee

    2016-07-01

    Full Text Available It has been reported that barley leaves possess beneficial properties such as antioxidant, hypolipidemic, antidepressant, and antidiabetic. Interestingly, barley sprouts contain a high content of saponarin, which showed both anti-inflammatory and antioxidant activities. In this study, we evaluated the effect of barley sprouts on alcohol-induced liver injury mediated by inflammation and oxidative stress. Raw barley sprouts were extracted, and quantitative and qualitative analyses of its components were performed. The mice were fed a liquid alcohol diet with or without barley sprouts for four weeks. Lipopolysaccharide (LPS-stimulated RAW 264.7 cells were used to study the effect of barley sprouts on inflammation. Alcohol intake for four weeks caused liver injury, evidenced by an increase in serum alanine aminotransferase and aspartate aminotransferase activities and tumor necrosis factor (TNF-α levels. The accumulation of lipid in the liver was also significantly induced, whereas the glutathione (GSH level was reduced. Moreover, the inflammation-related gene expression was dramatically increased. All these alcohol-induced changes were effectively prevented by barley sprouts treatment. In particular, pretreatment with barley sprouts significantly blocked inducible nitric oxide synthase (iNOS and cyclooxygenase (COX-2 expression in LPS-stimulated RAW 264.7. This study suggests that the protective effect of barley sprouts against alcohol-induced liver injury is potentially attributable to its inhibition of the inflammatory response induced by alcohol.

  2. Archaeogenetic evidence of ancient nubian barley evolution from six to two-row indicates local adaptation.

    Directory of Open Access Journals (Sweden)

    Sarah A Palmer

    Full Text Available BACKGROUND: Archaeobotanical samples of barley (Hordeum vulgare L. found at Qasr Ibrim display a two-row phenotype that is unique to the region of archaeological sites upriver of the first cataract of the Nile, characterised by the development of distinctive lateral bracts. The phenotype occurs throughout all strata at Qasr Ibrim, which range in age from 3000 to a few hundred years. METHODOLOGY AND FINDINGS: We extracted ancient DNA from barley samples from the entire range of occupancy of the site, and studied the Vrs1 gene responsible for row number in extant barley. Surprisingly, we found a discord between the genotype and phenotype in all samples; all the barley had a genotype consistent with the six-row condition. These results indicate a six-row ancestry for the Qasr Ibrim barley, followed by a reassertion of the two-row condition. Modelling demonstrates that this sequence of evolutionary events requires a strong selection pressure. CONCLUSIONS: The two-row phenotype at Qasr Ibrim is caused by a different mechanism to that in extant barley. The strength of selection required for this mechanism to prevail indicates that the barley became locally adapted in the region in response to a local selection pressure. The consistency of the genotype/phenotype discord over time supports a scenario of adoption of this barley type by successive cultures, rather than the importation of new barley varieties associated with individual cultures.

  3. Assessment of the Seedling Reactions of Some Hulless Barley Genotypes to Drechslera teres f. maculata

    OpenAIRE

    Gerlegiz, Emine Tuba; KARAKAYA, Aziz; Celik Oguz, Arzu; MERT, Zafer; Sayim, İsmail; Ergun, Namuk; Aydogan, Sinan

    2015-01-01

    The seedling reactions of three barley cultivars, one hulless barley cultivar, two candidate hulless barley lines and nine hulless barley genotypes were determined under greenhouse conditions to ten isolates of Drechslera teres f. maculata, the causal agent of spot form of net blotch. Isolates were obtained from Ankara, Çankırı, Eskişehir, Kayseri, Konya and Şanlıurfa provinces. The reactions of the cultivars and hulless cultivar ranged between suscepible-resistant. The reactions of the hulle...

  4. The non-touching method of the malting barley quality evaluation

    Science.gov (United States)

    Raba, B.; Nowakowski, K.; Lewicki, A.; Przybył, K.; Zaborowicz, M.; Koszela, K.; Boniecki, P.; Mueller, W.

    2014-04-01

    The first important stage of the malt production processes is the malting barley quality evaluation. Presented project was focused on the visual features of malting barley grains. The principal aim was to elaborate complete methodology to determine the level of grains contamination. The article describes the mechanisms of choosing parameters which can distinguish useful for the malt production grains from defects and impurities. Original computer system 'Hordeum v 3.1' helped obtain graphical data from images of contaminated barley samples. Research carried out in this area can improve the quality evaluation process of malting barley.

  5. Investigations of barley stripe mosaic virus as a gene silencing vector in barley roots and in Brachypodium distachyon and oat

    Directory of Open Access Journals (Sweden)

    Nilsson Lena

    2010-11-01

    Full Text Available Abstract Background Gene silencing vectors based on Barley stripe mosaic virus (BSMV are used extensively in cereals to study gene function, but nearly all studies have been limited to genes expressed in leaves of barley and wheat. However since many important aspects of plant biology are based on root-expressed genes we wanted to explore the potential of BSMV for silencing genes in root tissues. Furthermore, the newly completed genome sequence of the emerging cereal model species Brachypodium distachyon as well as the increasing amount of EST sequence information available for oat (Avena species have created a need for tools to study gene function in these species. Results Here we demonstrate the successful BSMV-mediated virus induced gene silencing (VIGS of three different genes in barley roots, i.e. the barley homologues of the IPS1, PHR1, and PHO2 genes known to participate in Pi uptake and reallocation in Arabidopsis. Attempts to silence two other genes, the Pi transporter gene HvPht1;1 and the endo-β-1,4-glucanase gene HvCel1, in barley roots were unsuccessful, probably due to instability of the plant gene inserts in the viral vector. In B. distachyon leaves, significant silencing of the PHYTOENE DESATURASE (BdPDS gene was obtained as shown by photobleaching as well as quantitative RT-PCR analysis. On the other hand, only very limited silencing of the oat AsPDS gene was observed in both hexaploid (A. sativa and diploid (A. strigosa oat. Finally, two modifications of the BSMV vector are presented, allowing ligation-free cloning of DNA fragments into the BSMV-γ component. Conclusions Our results show that BSMV can be used as a vector for gene silencing in barley roots and in B. distachyon leaves and possibly roots, opening up possibilities for using VIGS to study cereal root biology and to exploit the wealth of genome information in the new cereal model plant B. distachyon. On the other hand, the silencing induced by BSMV in oat seemed too

  6. Assay of methylglyoxal and glyoxal and control of peroxidase interference.

    Science.gov (United States)

    Thornalley, Paul J; Rabbani, Naila

    2014-04-01

    Methylglyoxal and glyoxal are endogenous α-oxoaldehyde metabolites and substrates of the glyoxalase system. These and related α-oxoaldehydes are often determined in cell, tissue and body fluid samples by derivatization with 1,2-diaminobenzene and similar compounds. Peroxidase activity in physiological tissues is a potential interference in estimation of methylglyoxal and glyoxal as it catalyses the conversion of 1,2-diaminobenzene into trace amounts of these dicarbonyl metabolites. Residual peroxidase activity in deproteinized extracts is found to cause significant interference in methylglyoxal and glyoxal estimations. This interference is blocked by the addition of sodium azide in the derivatizing buffer. Estimates of methylglyoxal concentration thereby obtained are in keeping with those predicted by systems modelling of methylglyoxal glycation kinetics in situ. Blocking sample peroxidase activity is important to avoid overestimation in the measurement of glyoxal and methylglyoxal. A dicarbonyl assay protocol resistant to interferences is described in the present article.

  7. Horseradish and soybean peroxidases: comparable tools for alternative niches?

    Science.gov (United States)

    Ryan, Barry J; Carolan, Neil; O'Fágáin, Ciarán

    2006-08-01

    Horseradish and soybean peroxidases (HRP and SBP, respectively) are useful biotechnological tools. HRP is often termed the classical plant heme peroxidase and although it has been studied for decades, our understanding has deepened since its cloning and subsequent expression, enabling numerous mutational and protein engineering studies. SBP, however, has been neglected until recently, despite offering a real alternative to HRP: SBP actually outperforms HRP in terms of stability and is now used in numerous biotechnological applications, including biosensors. Review of both is timely. This article summarizes and discusses the main insights into the structure and mechanism of HRP, with special emphasis on HRP mutagenesis, and outlines its use in a variety of applications. It also reviews the current knowledge and applications to date of SBP, particularly biosensors. The final paragraphs speculate on the future of plant heme-based peroxidases, with probable trends outlined and explored.

  8. Polymerization of phenols catalyzed by peroxidase in nonaqueous media

    Energy Technology Data Exchange (ETDEWEB)

    Dordick, J.S.; Marletta, M.A.; Klibanov, A.M.

    1987-01-01

    Polymers produced by horseradish-peroxidase-catalyzed coupling of phenols have been explored as potential substitutes for phenol-formaldehyde resins. To overcome low substrate solubilities and product molecular weights in water, enzymatic polymerizations in aqueous-organic mixtures have been examined. Peroxidase vigorously polymerizes a number of phenols in mixtures of water with water-miscible solvents such as dioxane, acetone, dimethylformamide, and methyl formate with the solvent content up to 95%. As a result, various phenolic polymers with average molecular weights from 400 to 2.6 x 10/sup 4/ D were obtained depending on the reaction medium composition and the nature of the phenol. Peroxidase-catalyzed copolymerization of different phenols in 85% dioxane was demonstrated. Poly(p-phenylphenol) and poly(p-cresol) were enzymatically prepared on a gram scale. They had much higher melting points, and in addition, poly(p-phenylphenol) was found to have a much higher electrical conductivity than phenol-formaldehyde resins.

  9. Optimization of extracellular fungal peroxidase production by 2 Coprinus species.

    Science.gov (United States)

    Ikehata, Keisuke; Pickard, Michael A; Buchanan, Ian D; Smith, Daniel W

    2004-12-01

    Optimum culture conditions for the batch production of extracellular peroxidase by Coprinus cinereus UAMH 4103 and Coprinus sp. UAMH 10067 were explored using 2 statistical experimental designs, including 2-level, 7-factor fractional factorial design and 2-factor central composite design. Of the 7 factors examined in the screening study, the concentrations of carbon (glucose) and nitrogen (peptone or casitone) sources showed significant effects on the peroxidase production by Coprinus sp. UAMH 10067. The optimum glucose and peptone concentrations were determined as 2.7% and 0.8% for Coprinus sp. UAMH 10067, and 2.9% and 1.4% for C. cinereus UAMH 4103, respectively. Under the optimized culture condition the maximum peroxidase activity achieved in this study was 34.5 U x mL(-1) for Coprinus sp. UAMH 10067 and 68.0 U x mL(-1) for C. cinereus UAMH 4103, more than 2-fold higher than the results of previous studies.

  10. Fluorescence Spectra and Enzymatic Property of Hemoglobin as Mimetic Peroxidase

    Institute of Scientific and Technical Information of China (English)

    Li De-jia; Li Hai-cheng; Zou Guo-lin

    2003-01-01

    Intrinsic fluorescence emission maxima of hemoglobin(Hb) was investigated in relation to peroxidase property of Hb. The peroxidase activity of Hb was based on its catalytic activity for oxidation of o-phenylenediamine by hydrogen peroxide. Hb was treated in the condition (temperature,ethanol and salt) that tetramer-dimer equilibrium of Hb is shifted to the dimer state and its fluorescence spectrum was measured. When Hb treated in temperature (60-70 ℃ ), ethanol concentration (60 %-70 % ) and NaCl concentration (2.5-3.0 mol/L), the fluorescence emission maxima of Hb shifted towards red wavelength and its activity decreased quickly.Experimental results revealed that the activity and stability of Hb as mimetic peroxidase was closely relative to the hydrophobic environment of active center of Hb, and when Hb (FeⅡ) converted into met Hb (FeⅢ ), its activity was 1. 6times as much as that of Hb.

  11. Fluorescence Spectra and Enzymatic Property of Hemoglobin as Mimetic Peroxidase

    Institute of Scientific and Technical Information of China (English)

    LiDe-jia; LiHai-cheng; ZouGuo-lin

    2003-01-01

    Intrinsic fluorescence emission maxima of hemo-lobin(Hb) was investigated in relation to peroxidase property of Hb. The peroxidase activity of Hb was based on its catalytic activity for oxidation of o-phenylenediamine by hydrogen peroxide. Hb was treated in the condition (temperature,ethanol and salt) that tetramer-dimer equilibrium of Hb is shifted to the dimer state and its fluorescence spectrum was measured. When Hb treated in temperature (60-70 ℃), ethanol concentration (60%-70%) and NaCl concentration (2. 5-3.0 mol/L), the fluorescence emission maxima of Hb shifted towards red wavelength and its activity decreased quickly.Experimental results revealed that the activity and stability of Hb as mimetic peroxidase was closely relative to the hydrophobic environment of active center of Hb, and when Hb (FeⅡ) converted into met Hb (FeⅢ ), its activity was 1. 6 times as much as that of Hb.

  12. Suppression of Zn stress on barley by irradiated chitosan

    Energy Technology Data Exchange (ETDEWEB)

    Nagasawa, N.; Mitomo, H. [Gunma Univ., Faculty of Engineering, Department of Biological and Chemical Engineering, Kiryu, Gunma (Japan); Ha, P.T.L. [Nuclear Research Institute, Dalat (Viet Nam); Watanabe, S.; Ito, T.; Takeshita, H.; Yoshii, F.; Kume, T. [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

    2001-03-01

    Chitosan was irradiated up to 1000 kGy in solid state. Irradiation of chitosan caused the reduction of molecular weight. The molecular weight of the chitosan reduced from ca. 4 x 10{sup 5} to ca. 6 x 10{sup 3} by irradiation at 1000 kGy. For the barley growth promotion, irradiated chitosan showed the significant effect and 1000 kGy irradiated chitosan improved 20% of growth. Using the positron emitting tracer imaging system (PETIS), the effect of chitosan on uptake and transportation of {sup 62}Zn in barley were investigated. It was found that the transportation of Zn from root to shoot and the damage of plant by Zn were suppressed with irradiated chitosan. (author)

  13. In vitro fermentation of ten cultivars of barley silage

    Directory of Open Access Journals (Sweden)

    Federico Infascelli

    2010-01-01

    Full Text Available The fermentation characteristics of whole-crop barley silages from ten different cultivars were evaluated by the in vitro gas production technique. The organic matter degradability of barley silage (62.9% in average was comparable to those reported in our previous trials for oat (59.7% and sorghum silages (65.5%; while the maximum gas production rate (5.38 ml/h in average was slightly lower respect to oat (6.71 ml/h and sorghum silage (6.74 ml/h. The mean nutritive value (4.00 MJ/kg DM calculated on the basis of both chemical composition and in vitro fermentation data was comparable to that (4.16 MJ/kg DM obtained in our previous research performed on corn silage, from crop sowed in the same area.

  14. Resistance in winter barley against Ramularia leaf spot

    DEFF Research Database (Denmark)

    Hjortshøj, Rasmus Lund

    Ramularia leaf spot is an emerging disease in barley caused by R. collo-cygni. At present little is known about the resistance mechanisms carried out by the host plant to avoid disease development. Nor is the lifecycle of the fungus or its populations structure fully understood. To gain insight...... into these aspects four experiments were set up; (1) a mapping experiment aiming at identifying QTL’s controlling disease levels under field conditions was conducted in two winterbarley populations. (2) a toxin assay testing the parental lines used in the mapping populations for response to Rubellin D was developed....... (3) microarray analysis of transcriptional response in barley to inoculation with R. collo-cygni was carried out, and finally the (4) population genetic structure of R. collo-cygni was assessed by AFLP and gene sequencing. Based on these experiments interaction was compared to the interactions of C...

  15. The Role of alpha-Glucosidase in Germinating Barley Grains

    DEFF Research Database (Denmark)

    Stanley, Duncan; Rejzek, Martin; Næsted, Henrik

    2011-01-01

    The importance of alpha-glucosidase in the endosperm starch metabolism of barley (Hordeum vulgare) seedlings is poorly understood. The enzyme converts maltose to glucose (Glc), but in vitro studies indicate that it can also attack starch granules. To discover its role in vivo, we took complementary...... chemical-genetic and reverse-genetic approaches. We identified iminosugar inhibitors of a recombinant form of an alpha-glucosidase previously discovered in barley endosperm (ALPHA-GLUCOSIDASE97 [HvAGL97]), and applied four of them to germinating grains. All four decreased the Glc-to-maltose ratio...... in the endosperm 10 d after imbibition, implying inhibition of maltase activity. Three of the four inhibitors also reduced starch degradation and seedling growth, but the fourth did not affect these parameters. Inhibition of starch degradation was apparently not due to inhibition of amylases. Inhibition...

  16. Barley grain enrichement with essential elements by agronomic biofortification

    Directory of Open Access Journals (Sweden)

    Dragičević Vesna D.

    2016-01-01

    Full Text Available Barley grain is rich in mineral nutrients, but their bioavailability to humans depends on antinutrients that restrain bioavailability and promoters that promote bioavailability. The aim of this study was to examine composition of barley grain, including phytate and phenolics as antinutrients, carotenoids and glutathione as promoters and mineral elements, such as Ca, Mg, Fe, Si, Zn and Mn influenced by various non-standard foliar fertilizers (Zircon, Chitosan, Siliplant, Propikonazole, including some hormonal growth-stimulators (Epin Extra, Benzyladenine, as potential biofortification measure. Chitosan increased glutathione concentration in grain. Unfavorable meteorological conditions were partly mitigated by application of Benzyladenine and Siliplant, reflected through increased potential bioavailability of P, Mg, Ca and Fe. [Projekat Ministarstva nauke Republike Srbije, br. TR-31037

  17. Pollen grain of barley (Hordeum vulgare L. - pattern of development

    Directory of Open Access Journals (Sweden)

    Maria Charzyńska

    2014-01-01

    Full Text Available Pollen development in barley follows the general pattern established for other species of Poaceae: 1 microspore division occurs at the vacuolate microspore stage with polarly located nucleus; 2 microspore mitosis is immediately followed by phragmoplast and cell plate formation; 3 in consequence or unequal microspore division, the generative cell, at first attached to the pollen wall, is separated from the vegetative cell by a callosic wall; 4 during the postmitotic two-cell stage of development, the vegetative nucleus migrates to the aperture pole and is followed by the generative cell that is detached and free of callose wall. In this position the generative cell divides into two sperm cells. These data do not confirm the interpretation of pollen grain development in barley given by Cass and Karas in Can. J. Bot. 53: 1051-1062, 1975.

  18. Environmental and transgene expression effects on the barley seed proteome

    DEFF Research Database (Denmark)

    Finnie, Christine; Steenholdt, T.; Noguera, O.R.;

    2004-01-01

    The barley (Hordeum vulgare) cultivar Golden Promise is no longer widely used for malting, but is amenable to transformation and is therefore a valuable experimental cultivar. Its characteristics include high salt tolerance, however it is also susceptible to several fungal pathogens. Proteome....... Eleven of these were identified by mass spectrometric peptide mass mapping, including an abundant chitinase implicated in defence against fungal pathogens and a small heat-shock protein. To enable a comparison with transgenic seed protein patterns, differences in spot patterns between field...... with extra nitrogen. Finally, the fate of transgene products in barley seeds was followed. Spots containing two green fluorescent protein constructs and the herbicide resistance marker phosphinothricin acetyltransferase were observed in 2D-gel patterns of transgenic seeds and identified by mass spectrometry...

  19. Effect of ozone pretreatment on hydrogen production from barley straw.

    Science.gov (United States)

    Wu, Jiangning; Ein-Mozaffari, Farhad; Upreti, Simant

    2013-09-01

    Application of ozone technology to lignocellulosic biohydrogen production was explored with a barley straw. Ozone pretreatment effectively degraded the straw lignin and increased reducing sugar yield. A simultaneous enzyme hydrolysis and dark fermentation experiment was conducted using a mixed anaerobic consortium together with saccharification enzymes. Both untreated and ozonated samples produced hydrogen. Compared to the untreated group, hydrogen produced by the groups ozonated for 15, 30, 45 and 90 min increased 99%, 133%, 166% and 94%, respectively. Some inhibitory effect on hydrogen production was observed with the samples ozonated for 90 min, and the inhibition was on the fermentative microorganisms, not the saccharification enzymes. These results demonstrate that production of biohydrogen from barley straw, a lignocellulosic biomass, can be significantly enhanced by ozone pretreatment.

  20. Integration of the barley genetic and seed proteome maps for chromosome 1H, 2H, 3H, 5H and 7H.

    Science.gov (United States)

    Finnie, Christine; Bagge, Merethe; Steenholdt, Torben; Østergaard, Ole; Bak-Jensen, Kristian Sass; Backes, Gunter; Jensen, Anaïs; Giese, Henriette; Larsen, Jørgen; Roepstorff, Peter; Svensson, Birte

    2009-02-01

    Two-dimensional gel electrophoresis was used to screen spring barley cultivars for differences in seed protein profiles. In parallel, 72 microsatellite (simple sequence repeat (SSR)) markers and 11 malting quality parameters were analysed for each cultivar. Over 60 protein spots displayed cultivar variation, including peroxidases, serpins and proteins with unknown functions. Cultivars were clustered based on the spot variation matrix. Cultivars with superior malting quality grouped together, indicating malting quality to be more closely correlated with seed proteomes than with SSR profiles. Mass spectrometry showed that some spot variations were caused by amino acid differences encoded by single nucleotide polymorphisms (SNPs). Coding SNPs were validated by mass spectrometry, expressed sequence tag and 2D gel data. Coding SNPs can alter function of affected proteins and may thus represent a link between cultivar traits, proteome and genome. Proteome analysis of doubled haploid lines derived from a cross between a malting (Scarlett) and a feed cultivar (Meltan) enabled genetic localisation of protein phenotypes represented by 48 spot variations, involving e.g. peroxidases, serpins, alpha-amylase/trypsin inhibitors, peroxiredoxin and a small heat shock protein, in relation to markers on the chromosome map.

  1. Targeting the active site of the placental isozyme of alkaline phosphatase by phage-displayed scFv antibodies selected by a specific uncompetitive inhibitor

    Directory of Open Access Journals (Sweden)

    Kala Mrinalini

    2005-12-01

    Full Text Available Abstract Background The isozymes of alkaline phosphatase, the tissue non-specific, intestinal and placental, have similar properties and a high degree of identity. The placental isozyme (PLAP is an oncofetal antigen expressed in several malignancies including choriocarcinoma, seminoma and ovarian carcinoma. We had earlier attempted to isolate PLAP-specific scFv from a synthetic human immunoglobulin library but were unable to do so, presumably because of the similarity between the isozymes. In this work, we have employed a PLAP-specific uncompetitive inhibitor, L-Phe-Gly-Gly, to select isozyme specific scFvs. An uncompetitive inhibitor binds to the enzyme in the presence of substrate and stabilizes the enzyme-substrate complex. Several uncompetitive inhibitors have varying degrees of isozyme specificity for human alkaline phosphatase isozymes. A specific uncompetitive inhibitor would be able to unmask conformational differences between the otherwise very similar molecules. Also, such inhibitors would be directed to regions at/close to the active site of the enzyme. In this work, the library was first incubated with PLAP and the bound clones then eluted by incubation with L-Phe-Gly-Gly along with the substrate, para-nitro phenyl phosphate (pNPP. The scFvs were then studied with regard to the biochemical modulation of their binding, isozyme specificity and effect on enzyme activity. Results Of 13 clones studied initially, the binding of 9 was inhibited by L-Phe-Gly-Gly (with pNPP and 2 clones were inhibited by pNPP alone. Two clones had absolute and 2 clones had partial specificity to PLAP. Two clones were cross-reactive with only one other isozyme. Three scFv clones, having an accessible His6-tag, were purified and studied for their modulation of enzyme activity. All the three scFvs inhibited PLAP activity with the kinetics of competitive inhibition. Cell ELISA could demonstrate binding of the specific scFvs to the cell surface expressed PLAP

  2. Inorganic chemistry of defensive peroxidases in the human oral cavity.

    Science.gov (United States)

    Ashby, M T

    2008-10-01

    The innate host response system is comprised of various mechanisms for orchestrating host response to microbial infection of the oral cavity. The heterogeneity of the oral cavity and the associated microenvironments that are produced give rise to different chemistries that affect the innate defense system. One focus of this review is on how these spatial differences influence the two major defensive peroxidases of the oral cavity, salivary peroxidase (SPO) and myeloperoxidase (MPO). With hydrogen peroxide (H(2)O(2)) as an oxidant, the defensive peroxidases use inorganic ions to produce antimicrobials that are generally more effective than H(2)O(2) itself. The concentrations of the inorganic substrates are different in saliva vs. gingival crevicular fluid (GCF). Thus, in the supragingival regime, SPO and MPO work in unison for the exclusive production of hypothiocyanite (OSCN(-), a reactive inorganic species), which constantly bathes nascent plaques. In contrast, MPO is introduced to the GCF during inflammatory response, and in that environment it is capable of producing hypochlorite (OCl(-)), a chemically more powerful oxidant that is implicated in host tissue damage. A second focus of this review is on inter-person variation that may contribute to different peroxidase function. Many of these differences are attributed to dietary or smoking practices that alter the concentrations of relevant inorganic species in the oral cavity (e.g.: fluoride, F(-); cyanide, CN(-); cyanate, OCN(-); thiocyanate, SCN(-); and nitrate, NO(3)(-)). Because of the complexity of the host and microflora biology and the associated chemistry, it is difficult to establish the significance of the human peroxidase systems during the pathogenesis of oral diseases. The problem is particularly complex with respect to the gingival sulcus and periodontal pockets (where the very different defensive stratagems of GCF and saliva co-mingle). Despite this complexity, intriguing in vitro and in vivo

  3. Nutritional assessment of barley, talbina and their germinated products

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    Mohamed kamal El-Sayed Youssef

    2013-02-01

    Full Text Available Talbina is a food product with high potential applications as a functional food. Talbina was prepared from two barley varieties namely: Giza126 and Giza130 by adding whole barley flour to water (1:10 w/v and (1:5 w/v for germinated barley then heating at  80° C for 5 minutes with continuous stirring until reaching a porridge like texture. The present investigation was carried out in an attempt to clearly the nutritional assessment of talbina as a functional food. The study included the determination of gross chemical composition, caloric value, mineral composition, vitamins composition and the amino acids composition. Meanwhile, computation of the chemical scores (CS and A/E ratios were carried out for raw, germinated barley, talbina, germinated talbina and commercial talbina. The data revealed that protein content of the all raw studied and processing treatments ranged from 8.75-18.34g/100g on dry weight basis. Besides, the all treatments recorded rather slight decrease in crude fat content. Likewise, ash and carbohydrates ranged between 2.29-2.86 and 73.40-82.66%, respectively. Whereas crude fiber had an increase after treatments and it ranged from 3.83-4.37%. On the other hand by making talbina iron, manganese, copper and zinc increased especially zinc, which recorded higher value than that recommended daily. Furthermore, germinated talbina130 recorded the highest amounts of vitamins B2, Nicotinic acid, B6 and folic acid. Moreover, the present study indicated that phenylalanine was the highest essential amino acid, followed by leucine.

  4. Drivers of phosphorus uptake by barley following secondary resource application

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    Eva eBrod

    2016-05-01

    Full Text Available Minable rock phosphate is a finite resource. Replacing mineral phosphorus (P fertilizer with P-rich secondary resources is one way to manage P more efficiently, but the importance of physicochemical and microbial soil processes induced by secondary resources for plant P uptake are still poorly understood. Using radioactive labelling techniques, the fertilization effects of dairy manure, fish sludge, meat bone meal and wood ash were studied as P uptake by barley after 44 days and compared with those of water-soluble mineral P (MinP and an unfertilized control (NoP in a pot experiment with an agricultural soil containing little available P at two soil pH levels, approximately pH 5.3 (unlimed soil and pH 6.2 (limed soil. In a parallel incubation experiment, the effects of the secondary resources on physicochemical and microbial soil processes were studied. The results showed that the relative agronomic efficiency compared with MinP decreased in the order: manure ≥ fish sludge ≥ wood ash ≥ meat bone meal. The solubility of inorganic P in secondary resources was the main driver for P uptake by barley (Hordeum vulgare. The effects of secondary resources on physicochemical and microbial soil processes were of little overall importance. Application of organic carbon with manure resulted in microbial P immobilisation and decreased uptake by barley of P derived from the soil. On both soils, P uptake by barley was best explained by a positive linear relationship with the H2O + NaHCO3-soluble inorganic P fraction in fertilizers, or by a linear negative relationship with the HCl-soluble inorganic P fraction in fertilizers.

  5. Golgi localized barley MTP8 proteins facilitate Mn transport.

    Science.gov (United States)

    Pedas, Pai; Schiller Stokholm, Michaela; Hegelund, Josefine Nymark; Ladegård, Anne Hald; Schjoerring, Jan Kofod; Husted, Søren

    2014-01-01

    Many metabolic processes in plants are regulated by manganese (Mn) but limited information is available on the molecular mechanisms controlling cellular Mn homeostasis. In this study, a yeast assay was used to isolate and characterize two genes, MTP8.1 and MTP8.2, which encode membrane-bound proteins belonging to the cation diffusion facilitator (CDF) family in the cereal species barley (Hordeum vulgare). Transient expression in onion epidermal cells showed that MTP8.1 and MTP8.2 proteins fused to the green fluorescent protein (GFP) are localized to Golgi. When heterologously expressed in yeast, MTP8.1 and MTP8.2 were found to be Mn transporters catalysing Mn efflux in a similar manner as the Golgi localized endogenous yeast protein Pmr1p. The level of MTP8.1 transcripts in barley roots increased with external Mn supply ranging from deficiency to toxicity, while MTP8.2 transcripts decreased under the same conditions, indicating non-overlapping functions for the two genes. In barley leaves, the expression of both MTP8 genes declined in response to toxic Mn additions to the roots suggesting a role in ensuring proper delivery of Mn to Golgi. Based on the above we suggest that barley MTP8 proteins are involved in Mn loading to the Golgi apparatus and play a role in Mn homeostasis by delivering Mn to Mn-dependent enzymes and/or by facilitating Mn efflux via secretory vesicles. This study highlights the importance of MTP transporters in Mn homeostasis and is the first report of Golgi localized Mn2+ transport proteins in a monocot plant species.

  6. Genomic Regions Influencing Seminal Root Traits in Barley

    Directory of Open Access Journals (Sweden)

    Hannah Robinson

    2016-03-01

    Full Text Available Water availability is a major limiting factor for crop production, making drought adaptation and its many component traits a desirable attribute of plant cultivars. Previous studies in cereal crops indicate that root traits expressed at early plant developmental stages, such as seminal root angle and root number, are associated with water extraction at different depths. Here, we conducted the first study to map seminal root traits in barley ( L.. Using a recently developed high-throughput phenotyping method, a panel of 30 barley genotypes and a doubled-haploid (DH population (ND24260 × ‘Flagship’ comprising 330 lines genotyped with diversity array technology (DArT markers were evaluated for seminal root angle (deviation from vertical and root number under controlled environmental conditions. A high degree of phenotypic variation was observed in the panel of 30 genotypes: 13.5 to 82.2 and 3.6 to 6.9° for root angle and root number, respectively. A similar range was observed in the DH population: 16.4 to 70.5 and 3.6 to 6.5° for root angle and number, respectively. Seven quantitative trait loci (QTL for seminal root traits (root angle, two QTL; root number, five QTL were detected in the DH population. A major QTL influencing both root angle and root number (/ was positioned on chromosome 5HL. Across-species analysis identified 10 common genes underlying root trait QTL in barley, wheat ( L., and sorghum [ (L. Moench]. Here, we provide insight into seminal root phenotypes and provide a first look at the genetics controlling these traits in barley.

  7. Golgi localized barley MTP8 proteins facilitate Mn transport.

    Directory of Open Access Journals (Sweden)

    Pai Pedas

    Full Text Available Many metabolic processes in plants are regulated by manganese (Mn but limited information is available on the molecular mechanisms controlling cellular Mn homeostasis. In this study, a yeast assay was used to isolate and characterize two genes, MTP8.1 and MTP8.2, which encode membrane-bound proteins belonging to the cation diffusion facilitator (CDF family in the cereal species barley (Hordeum vulgare. Transient expression in onion epidermal cells showed that MTP8.1 and MTP8.2 proteins fused to the green fluorescent protein (GFP are localized to Golgi. When heterologously expressed in yeast, MTP8.1 and MTP8.2 were found to be Mn transporters catalysing Mn efflux in a similar manner as the Golgi localized endogenous yeast protein Pmr1p. The level of MTP8.1 transcripts in barley roots increased with external Mn supply ranging from deficiency to toxicity, while MTP8.2 transcripts decreased under the same conditions, indicating non-overlapping functions for the two genes. In barley leaves, the expression of both MTP8 genes declined in response to toxic Mn additions to the roots suggesting a role in ensuring proper delivery of Mn to Golgi. Based on the above we suggest that barley MTP8 proteins are involved in Mn loading to the Golgi apparatus and play a role in Mn homeostasis by delivering Mn to Mn-dependent enzymes and/or by facilitating Mn efflux via secretory vesicles. This study highlights the importance of MTP transporters in Mn homeostasis and is the first report of Golgi localized Mn2+ transport proteins in a monocot plant species.

  8. Improved avidin-biotin-peroxidase complex (ABC) staining.

    Science.gov (United States)

    Cattoretti, G; Berti, E; Schiró, R; D'Amato, L; Valeggio, C; Rilke, F

    1988-02-01

    A considerable intensification of the avidin-biotin-peroxidase complex staining system (ABC) was obtained by sequentially overlaying the sections to be immunostained with an avidin-rich and a biotin-rich complex. Each sequential addition contributed to the deposition of horseradish peroxidase on the immunostained site and allowed the subsequent binding of a complementary complex. With this technique a higher dilution of the antisera could be used and minute amounts of antigen masked by the fixative could be demonstrated on paraffin sections.

  9. Unique and Conserved Features of the Barley Root Meristem

    Directory of Open Access Journals (Sweden)

    Gwendolyn K. Kirschner

    2017-07-01

    Full Text Available Plant root growth is enabled by root meristems that harbor the stem cell niches as a source of progenitors for the different root tissues. Understanding the root development of diverse plant species is important to be able to control root growth in order to gain better performances of crop plants. In this study, we analyzed the root meristem of the fourth most abundant crop plant, barley (Hordeum vulgare. Cell division studies revealed that the barley stem cell niche comprises a Quiescent Center (QC of around 30 cells with low mitotic activity. The surrounding stem cells contribute to root growth through the production of new cells that are displaced from the meristem, elongate and differentiate into specialized root tissues. The distal stem cells produce the root cap and lateral root cap cells, while cells lateral to the QC generate the epidermis, as it is typical for monocots. Endodermis and inner cortex are derived from one common initial lateral to the QC, while the outer cortex cell layers are derived from a distinct stem cell. In rice and Arabidopsis, meristem homeostasis is achieved through feedback signaling from differentiated cells involving peptides of the CLE family. Application of synthetic CLE40 orthologous peptide from barley promotes meristem cell differentiation, similar to rice and Arabidopsis. However, in contrast to Arabidopsis, the columella stem cells do not respond to the CLE40 peptide, indicating that distinct mechanisms control columella cell fate in monocot and dicot plants.

  10. Genetic analysis of aluminum tolerance in Brazilian barleys

    Directory of Open Access Journals (Sweden)

    Minella Euclydes

    2002-01-01

    Full Text Available Aluminum (Al toxicity is a major factor limiting barley growth in acid soils, and genotypes with adequate level of tolerance are needed for improving barley adaptation in Brazil. To study the inheritance of Al tolerance in Brazilian barleys, cultivars Antarctica 1, BR 1 and FM 404 were crossed to sensitive Kearney and PFC 8026, and intercrossed. Parental, F1, F2 and F6 generations were grown in nutrient solution containing 0.03, 0.05 and 0.07 mM of Al and classified for tolerance by the root tip hematoxylin staining assay. Tolerant by sensitive F2 progenies segregated three tolerant to one sensitive, fitting the 3:1 ratio expected for a single gene. The F6 populations segregated one tolerant to one sensitive also fitting a monogenic ratio. The F2 seedlings from crosses among tolerant genotypes scored the same as the parents. Since the population size used would allow detection of recombination as low as 7%, the complete absence of Al sensitive recombinants suggests that tolerance in these cultivars is most probably, controlled by the same gene. Thus, the potential for improving Al tolerance through recombination of these genotypes is very low and different gene sources should be evaluated.

  11. Pysicochemical properties of Tibetan hull-less barley starch.

    Science.gov (United States)

    Yangcheng, Hanyu; Gong, Lingxiao; Zhang, Ying; Jane, Jay-lin

    2016-02-10

    Objectives of this study were to (1) determine the starch physicochemical properties of two commercial Tibetan hull-less barley varieties, Beiqing (BQ) and Kangqing (KQ); and (2) understand the relationship between unique properties of the starches, their structures, and impacts of growing conditions. The BQ barleys were grown at a location with lower temperature and less rainfall compared with the KQ barleys. The BQ starches showed significantly lower onset-gelatinization temperature (54.1-54.9 °C), larger gelatinization-temperature range (9.4-10.6 °C), and higher peak-viscosities (138.9-153.9RVU) than the KQ starches (55.1-56.1 °C, 7.4-8.8 °C, and 63.4-64.7RVU, respectively). After a treatment with 2% sodium-dodecyl-sulphate solution, the KQ starches showed substantially greater increases in peak viscosities than the BQ starches. Annealing of starch and enhanced amylose-lipid complex formation, resulting from higher growing temperature during the development of the KQ starches, likely contributed to the differences in thermal and pasting properties between the BQ and KQ starches.

  12. Unique and Conserved Features of the Barley Root Meristem.

    Science.gov (United States)

    Kirschner, Gwendolyn K; Stahl, Yvonne; Von Korff, Maria; Simon, Rüdiger

    2017-01-01

    Plant root growth is enabled by root meristems that harbor the stem cell niches as a source of progenitors for the different root tissues. Understanding the root development of diverse plant species is important to be able to control root growth in order to gain better performances of crop plants. In this study, we analyzed the root meristem of the fourth most abundant crop plant, barley (Hordeum vulgare). Cell division studies revealed that the barley stem cell niche comprises a Quiescent Center (QC) of around 30 cells with low mitotic activity. The surrounding stem cells contribute to root growth through the production of new cells that are displaced from the meristem, elongate and differentiate into specialized root tissues. The distal stem cells produce the root cap and lateral root cap cells, while cells lateral to the QC generate the epidermis, as it is typical for monocots. Endodermis and inner cortex are derived from one common initial lateral to the QC, while the outer cortex cell layers are derived from a distinct stem cell. In rice and Arabidopsis, meristem homeostasis is achieved through feedback signaling from differentiated cells involving peptides of the CLE family. Application of synthetic CLE40 orthologous peptide from barley promotes meristem cell differentiation, similar to rice and Arabidopsis. However, in contrast to Arabidopsis, the columella stem cells do not respond to the CLE40 peptide, indicating that distinct mechanisms control columella cell fate in monocot and dicot plants.

  13. The Metabolic Signature of Biomass Formation in Barley.

    Science.gov (United States)

    Ghaffari, Mohammad R; Shahinnia, Fahimeh; Usadel, Björn; Junker, Björn; Schreiber, Falk; Sreenivasulu, Nese; Hajirezaei, Mohammad R

    2016-09-01

    The network analysis of genome-wide transcriptome responses, metabolic signatures and enzymes' relationship to biomass formation has been studied in a diverse panel of 12 barley accessions during vegetative and reproductive stages. The primary metabolites and enzymes involved in central metabolism that determine the accumulation of shoot biomass at the vegetative stage of barley development are primarily being linked to sucrose accumulation and sucrose synthase activity. Interestingly, the metabolic and enzyme links which are strongly associated with biomass accumulation during reproductive stages are related to starch accumulation and tricarboxylic acid (TCA) cycle intermediates citrate, malate, trans-aconitate and isocitrate. Additional significant associations were also found for UDP glucose, ATP and the amino acids isoleucine, valine, glutamate and histidine during the reproductive stage. A network analysis resulted in a combined identification of metabolite and enzyme signatures indicative for grain weight accumulation that was correlated with the activity of ADP-glucose pyrophosphorylase (AGPase), a rate-limiting enzyme involved in starch biosynthesis, and with that of alanine amino transferase involved in the synthesis of storage proteins. We propose that the mechanism related to vegetative and reproductive biomass formation vs. seed biomass formation is being linked to distinct fluxes regulating sucrose, starch, sugars and amino acids as central resources. These distinct biomarkers can be used to engineer biomass production and grain weight in barley.

  14. Effects of ethylene on root elongation in barley and rice

    Energy Technology Data Exchange (ETDEWEB)

    John, A.; Hall, M.A.; Crossett, R.N.

    1972-01-01

    Experiments were performed to determine the effects of rice and barley to growth inhibition by ethylene. The mechanism of growth inhibition was investigated at the cellular level and a detailed comparison was made between the responses of the two species. The following measurements were made on intact plants in short (up to 200 minutes), medium (up to 3 days) or long (up to 10 days) experiments: the rate of extension growth of main root axes; the final cell length and number of elongating cells produced; and the extensibility of the apical growing region. Results indicate that the effects of ethylene on the elongation of roots of rice and barley plants are different. In barley there is a rapid inhibition of root extension which persists with prolonged exposure to the gas but with little effect on the production of growing cells. However, rice roots exhibit no rapid growth inhibition response, but a reduction does occur after prolonged exposure. Low concentrations promote extension rice roots. The inhibition of root growth is reflected in a reduced extensibility of the apical growing region.

  15. Nitrate Uptake into Barley (Hordeum vulgare) Plants 1

    Science.gov (United States)

    Deane-Drummond, Celia E.; Glass, Anthony D. M.

    1982-01-01

    Evidence is presented that chlorate is an extremely good analog for nitrate during nitrate uptake by intact barley (Hordeum vulgare cv. Fergus) roots. The depletion of ClO3− or NO3− from uptake media over 2 to 6 hours by seedlings was found to be dependent on combined NO3− plus ClO3− concentrations, and total anion uptake was equivalent at different NO3−/ClO3− ratios. After loading barley seedlings with 36ClO3− for 6 hours, kinetic parameters were derived from the analysis of efflux of [36Cl] chlorate into unlabeled solution. On the basis of this analysis, the half times for exchange for the cytoplasmic and vacuolar phases were 17 minutes and 20 hours, respectively. Data pooled from a number of different experiments were used to calculate kinetic constants (Km and Vmax) for 36ClO3− influx into barley roots at different external ClO3−/NO3− ratios, using short (10 minutes) influx times. There appeared to be no discrimination by the root cells between ClO3− and NO3−. Lineweaver-Burk analysis of the interaction between nitrate and chlorate were characteristic of competitive inhibition at low nitrate concentrations (0-0.5 mm). At higher concentrations, in the range of >1 mm, similar interactions between these ions were evident. PMID:16662478

  16. Screening of the aerodynamic and biophysical properties of barley malt

    Science.gov (United States)

    Ghodsvali, Alireza; Farzaneh, Vahid; Bakhshabadi, Hamid; Zare, Zahra; Karami, Zahra; Mokhtarian, Mohsen; Carvalho, Isabel. S.

    2016-10-01

    An understanding of the aerodynamic and biophysical properties of barley malt is necessary for the appropriate design of equipment for the handling, shipping, dehydration, grading, sorting and warehousing of this strategic crop. Malting is a complex biotechnological process that includes steeping; germination and finally, the dehydration of cereal grains under controlled temperature and humidity conditions. In this investigation, the biophysical properties of barley malt were predicted using two models of artificial neural networks as well as response surface methodology. Stepping time and germination time were selected as the independent variables and 1 000 kernel weight, kernel density and terminal velocity were selected as the dependent variables (responses). The obtained outcomes showed that the artificial neural network model, with a logarithmic sigmoid activation function, presents more precise results than the response surface model in the prediction of the aerodynamic and biophysical properties of produced barley malt. This model presented the best result with 8 nodes in the hidden layer and significant correlation coefficient values of 0.783, 0.767 and 0.991 were obtained for responses one thousand kernel weight, kernel density, and terminal velocity, respectively. The outcomes indicated that this novel technique could be successfully applied in quantitative and qualitative monitoring within the malting process.

  17. The white barley mutant albostrians shows a supersusceptible but symptomless interaction phenotype with the hemibiotrophic fungus Bipolaris sorokiniana.

    Science.gov (United States)

    Schäfer, Patrick; Hückelhoven, Ralph; Kogel, Karl-Heinz

    2004-04-01

    Bipolaris sorokiniana (teleomorph: Cochliobolus sativus) is a cereal pathogen of increasing global concern, with most significance in Asiatic cropping systems. In order to gain insight into the mechanism of host resistance, we studied fungal development on the supersusceptible barley mutant albostrians and its parent cv. Haisa. A microscopic dissection of early fungal growth on Haisa and green albostrians leaves revealed a distinct epidermis-localized biotrophic and a mesophyll-based necrotrophic phase. White, green, and striped white-green albostrians leaves showed extreme differences in disease development. When comparing cellular defense responses, we found restriction of fungal spreading after successful infection of host mesophyll tissue to be the most important mechanism limiting outbreak of the disease. Colonization of susceptible green leaves, but not extreme colonization of supersusceptible white albostrians leaves, was associated with macroscopically visible lesion formation and mesophyll accumulation of hydrogen peroxide (H2O2), implying a symptomless growth of the pathogen in supersusceptible host tissue. In contrast, early epidermal papilla-based resistance was closely linked to H2O2 accumulation in all leaf types. In white leaves, ascorbate peroxidase (APX), glutathione-S-transferase (GST), and the cell death regulator Bax-inhibitor-1 (BI-1) showed a stronger constitutive or pathogen responsive activation, whereas glycolate oxidase (GLOX) and catalase (CAT2) expression was stronger in green leaves. We discuss supersusceptibility and symptomless growth on the basis of the histochemical and the gene expression data.

  18. Activity of the C-terminal-dependent vacuolar sorting signal of horseradish peroxidase C1a is enhanced by its secondary structure.

    Science.gov (United States)

    Matsui, Takeshi; Tabayashi, Ayako; Iwano, Megumi; Shinmyo, Atsuhiko; Kato, Ko; Nakayama, Hideki

    2011-02-01

    Plant class III peroxidase (PRX) catalyzes the oxidation and oxidative polymerization of a variety of phenolic compounds while reducing hydrogen peroxide. PRX proteins are classified into apoplast type and vacuole type based on the absence or the presence of C-terminal propeptides, which probably function as vacuolar sorting signals (VSSs). In this study, in order to improve our understanding of vacuole-type PRX, we analyzed regulatory mechanisms of vacuolar sorting of a model vacuole-type PRX, the C1a isozyme of horseradish (Armoracia rusticana) (HRP C1a). Using cultured transgenic tobacco cells and protoplasts derived from horseradish leaves, we characterized HRP C1a's VSS, which is a 15 amino acid C-terminal propeptide (C15). We found that the C-terminal hexapeptide of C15 (C6), which is well conserved among vacuole-type PRX proteins, forms the core of the C-terminal-dependent VSS. We also found that the function of C6 is enhanced by the remaining N-terminal part of C15 which probably folds into an amphiphilic α-helix.

  19. Peroxidase activity in Spondias dulcis = Atividade da peroxidase em Spondias dulcis

    Directory of Open Access Journals (Sweden)

    Lúcio Cardozo-Filho

    2010-10-01

    Full Text Available In this study, the best conditions to obtain crude extracts showingPeroxidase activity from Spondia dulcis (caja-mango were evaluated. Fresh fruits (25 g were blended in different sodium phosphate buffer (0.05 to 0.2 M with a pH varying from 3.0 to 9.0. The muddy material was centrifuged for 20 minutes. In order to improve POD activity, the crude extract was submitted to precipitation with ammonium sulfate at 90% saturation. This precipitated was re-suspended in sodium phosphate buffer 0.2 M pH 6.5 and then, optimum pH for activity assay (pH varying from 5.0 to 9.0 and thermal stability (exposure to different temperatures varying from 30 to 75ºC for periods between 0 to 15 minutes were determined. The best conditions for activity assay were in phosphate buffer 0.2 M at pH7.0. The results obtained for thermal inactivation study suggest that the heating at 75ºCfor 15 minutes inactivated 95% of initial POD activity.Foram avaliadas, neste trabalho, algumas condições para a obtenção de extratos brutos com atividade peroxidase de Spondias dulcis (cajá-manga. Frutas frescas (25 g foram trituradas com tampão fosfato de sódio (0,05 a 0,2 M em pHs diferentes (3,0 a 9,0. O material obtido foi centrifugado por 20 min. O extrato bruto foi submetido à precipitação com sulfato de amônio até 90% de saturação. Este precipitado foi ressuspenso em tampão fosfato de sódio 0,2 M pH 6,5 e, assim, o pH ótimo para o ensaio de atividade (pH que varia de 5,0 a 9,0 e a estabilidade térmica (exposição a temperaturas de 30, 60, 65, 70 e 75ºC por um período de 0 a 15 min. deste foram determinados. As melhores condições encontradas para o ensaio de atividade foram em tampão fosfato 0,2 M pH 7,0. Os resultados para a inativação térmica sugerem que o aquecimento a 75ºC por 15 mininativa 95% da atividade de POD inicial.

  20. Arabidopsis ATP A2 peroxidase. Expression and high-resolution structure of a plant peroxidase with implications for lignification

    DEFF Research Database (Denmark)

    Ostergaard, L; Teilum, K; Mirza, O;

    2000-01-01

    to be involved in lignin biosynthesis. Recently we isolated an extracellular anionic peroxidase, ATP A2, from rapidly lignifying Arabidopsis cell suspension culture and cloned its cDNA. Here we show that the Atp A2 promoter directs GUS reporter gene expression in lignified tissues of transgenic plants. Moreover......Lignins are phenolic biopolymers synthesized by terrestrial, vascular plants for mechanical support and in response to pathogen attack. Peroxidases have been proposed to catalyse the dehydrogenative polymerization of monolignols into lignins, although no specific isoenzyme has been shown......-coumaryl and coniferyl alcohols are preferred by ATP A2, while the oxidation of sinapyl alcohol will be sterically hindered in ATP A2 as well as in all other plant peroxidases due to an overlap with the conserved Pro-139. We suggest ATP A2 is involved in a complex regulation of the covalent cross-linking in the plant...

  1. Colonization history and introduction dynamics of capsella bursa-pastoris (Brassicaceae) in north america: isozymes and quantitative traits

    Science.gov (United States)

    Neuffer; Hurka

    1999-10-01

    Multilocus isozyme genotypic composition for aspartate aminotransferase (AAT), leucine aminopeptidase (LAP) and glutamate dehydrogenase (GDH) was studied for Capsella in the source continent, Europe (9000 plants from 593 populations), and in the colonized continent, North America (2700 plants from 88 populations). North America was depauperate in the number of genotypes (by approximately 50%), but in terms of frequencies, a few genotypes were common and shared by both continents. Although some, very rare, genotypes were, however, unique for North America, our data provided no evidence to indicate that the introduced gene pools were reconstructed on a multilocus genetic basis after introduction. Instead, they argued for a considerable number of independent introduction events. Geographical distribution patterns of multilocus genotypes in Europe and North America were pronounced and enabled us to trace the colonization history of Californian Capsella back to Spanish ancestral populations and those of temperate North America back to temperate European gene pools. A random-block field experiment with 14 Californian populations from different climatic regions revealed that variation patterns of quantitative traits reflect ecotypic variation, and the ecological amplitude of Capsella in North America is similar to that in Europe, which can be traced back to the introduction of preadapted genotypes. It appears that certain multilocus isozyme genotypes are associated with certain ecotypes. The variable European gene pool of Capsella was essentially introduced into North America without major genetic changes.

  2. Genetic diversity and differentiation through isozymes in natural populations of Pinus wallichiana A.B. Jacks (Blue Pine in India

    Directory of Open Access Journals (Sweden)

    Monnika Konnert

    2011-02-01

    Full Text Available Eighteen natural populations of Pinus wallichiana A.B. Jacks. (Blue Pine occurring in the Northwest Himalayas of India were studied on the basis of ten enzyme systems, representing 16 isozyme loci. Allele frequencies, genetic multiplicity, diversity and genetic differentiation values are presented. A marked allele frequency difference was found in different populations. The mean number of alleles per locus was 1.79, with a range from 1.56 to 2.06, and the gene pool diversity varied from 1.11 to 1.20. Out of 46 alleles 11 (23% appear to be rare. Of the 16 isozyme loci scored, 14 (87.5 % were polymorphic, based on the 99% criterion. Only the loci Pgm-A and Idh-A were found to be monomorphic in all the populations. The percentage of polymorphic loci ranged from 37.5% to 68.7575% among populations. Nei’s genetic distance ranged from 1.7% to 11%. The results show that natural populations of Pinus wallichiana in India contain an appreciableamount of genetic variation (P99 = 53.4%, Ho = 0.152, He = 0.14545 comparable to other pines (on average P99 = 52, H0 = 0.159, He = 0.159.

  3. Genetic diversity and differentiation through isozymes in natural populations of Pinus wallichiana A.B. Jacks (Blue Pine in India

    Directory of Open Access Journals (Sweden)

    Meena Bakshi

    2011-06-01

    Full Text Available Eighteen natural populations of Pinus wallichiana (Blue pine A.B. Jacks. occurring in Northwest Himalayas of India were studied on the basis of ten enzyme systems representing 16 isozyme loci. Allele frequencies, genetic multiplicity, diversity and genetic differentiation values are presented. A marked allele frequency difference was found in different populations. The mean number of alleles per locus was 1.79 with a range from 1.56 to 2.06 , the gene pool diversity varied from 1.11 to 1.20. Out of 46 alleles 11 (23% appear to be rare. Of the 16 isozyme loci scored 14 (87.5 % were polymorphic based on 99% criterion. Only the loci PGM-A and IDH-A were found to be monomorphic in all the populations. The percentage of polymorphic loci ranged from 37.5% to 68.7%. Nei's genetic distance ranged from 1.7% to 11%. The results show that natural populations of Pinus wallichiana in India contain appreciable amount of genetic variation (P99 = 53.4%, Ho = 0.152, He = 0.145 comparable to other pines (on average P99=52, H0=0.159, He = 0.159. 

  4. Molecular diversity of tuliposide B-converting enzyme in tulip (Tulipa gesneriana): identification of the root-specific isozyme.

    Science.gov (United States)

    Nomura, Taiji; Ueno, Ayaka; Ogita, Shinjiro; Kato, Yasuo

    2017-06-01

    6-Tuliposide B (PosB) is a glucose ester accumulated in tulip (Tulipa gesneriana) as a major secondary metabolite. PosB serves as the precursor of the antimicrobial lactone tulipalin B (PaB), which is formed by PosB-converting enzyme (TCEB). The gene TgTCEB1, encoding a TCEB, is transcribed in tulip pollen but scarcely transcribed in other tissues (e.g. roots) even though those tissues show high TCEB activity. This led to the prediction of the presence of a TCEB isozyme with distinct tissue specificity. Herein, we describe the identification of the TgTCEB-R gene from roots via native enzyme purification; this gene is a paralog of TgTCEB1. Recombinant enzyme characterization verified that TgTCEB-R encodes a TCEB. Moreover, TgTCEB-R was localized in tulip plastids, as found for pollen TgTCEB1. TgTCEB-R is transcribed almost exclusively in roots, indicating a tissue preference for the transcription of TCEB isozyme genes.

  5. The 5 Alpha-Reductase Isozyme Family: A Review of Basic Biology and Their Role in Human Diseases

    Directory of Open Access Journals (Sweden)

    Faris Azzouni

    2012-01-01

    Full Text Available Despite the discovery of 5 alpha-reduction as an enzymatic step in steroid metabolism in 1951, and the discovery that dihydrotestosterone is more potent than testosterone in 1968, the significance of 5 alpha-reduced steroids in human diseases was not appreciated until the discovery of 5 alpha-reductase type 2 deficiency in 1974. Affected males are born with ambiguous external genitalia, despite normal internal genitalia. The prostate is hypoplastic, nonpalpable on rectal examination and approximately 1/10th the size of age-matched normal glands. Benign prostate hyperplasia or prostate cancer does not develop in these patients. At puberty, the external genitalia virilize partially, however, secondary sexual hair remains sparse and male pattern baldness and acne develop rarely. Several compounds have been developed to inhibit the 5 alpha-reductase isozymes and they play an important role in the prevention and treatment of many common diseases. This review describes the basic biochemical properties, functions, tissue distribution, chromosomal location, and clinical significance of the 5 alpha-reductase isozyme family.

  6. Isozyme variation in four species of the Simulium perflavum species group (Diptera: Simuliidae from the Brazilian Amazon

    Directory of Open Access Journals (Sweden)

    Vera Margarete Scarpassa

    2003-01-01

    Full Text Available Electrophoretic studies of isozymes were done with four closely related species of the Simulium perflavum species group (Diptera: Simuliidade in the Brazilian Amazon, using last-instar larvae collected in the field. Ten enzymes were studied, which yielded 11 loci. Diagnostic loci were not found between Simulium maroniense cytotype D and Simulium rorotaense. Simulium maroniense and S. rorotaense differed from Simulium trombetense by two diagnostic loci (Me and Xdh, and Simulium perflavum differed from the other three species by four diagnostic loci (Me, Xdh, Mdh, and Got. The mean number of alleles per locus ranged from 1.30 to 2.30, the percentage of polymorphic loci ranged from 18.2 to 63.6% and the mean heterozygosity values observed ranged from 0.062 to 0.108. Genetic distances among the species ranged from 0.010 to 0.581. The lowest value was obtained between S. maroniense and S. rorotaense, and the highest between S. perflavum and S. trombetense. The genetic relationships among the four S. perflavum group species indicate that they are closely related. The high similarity at the isozyme level, allied to previous studies of morphology and polytene chromosomes, may suggest that the divergence time since the separation of S. maroniense and S. rorotaense is still too recent for diagnostic loci to have evolved.

  7. Self-Assembled Complexes of Horseradish Peroxidase with Magnetic Nanoparticles Showing Enhanced Peroxidase Activity

    KAUST Repository

    Corgié, Stéphane C.

    2012-02-15

    Bio-nanocatalysts (BNCs) consisting of horseradish peroxidase (HRP) self-assembled with magnetic nanoparticles (MNPs) enhance enzymatic activity due to the faster turnover and lower inhibition of the enzyme. The size and magnetization of the MNPs affect the formation of the BNCs, and ultimately control the activity of the bound enzymes. Smaller MNPs form small clusters with a low affinity for the HRP. While the turnover for the bound fraction is drastically increased, there is no difference in the H 2O 2 inhibitory concentration. Larger MNPs with a higher magnetization aggregate in larger clusters and have a higher affinity for the enzyme and a lower substrate inhibition. All of the BNCs are more active than the free enzyme or the MNPs (BNCs > HRP ≤laquo; MNPs). Since the BNCs show surprising resilience in various reaction conditions, they may pave the way towards new hybrid biocatalysts with increased activities and unique catalytic properties for magnetosensitive enzymatic reactions. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Observed and predicted changes over eight years in frequency of barley powdery mildew avirulent to spring barley in France and Denmark

    DEFF Research Database (Denmark)

    Bousset, L.; Hovmøller, M.S.; Caffier, V.

    2002-01-01

    Aerial populations of Blumeria graminis f.sp. hordei were studied in two French and two Danish regions from 1991 to 1999, at a time of year when only winter barley was present. A high frequency of genotypes not able to grow on the spring-sown crop of the previous growing season (denoted 'spring-a...... be predicted from the proportion of the barley area sown with winter barley, the use of resistance genes in the cultivars, the initial composition of the pathogen population, and hitch-hiking due to gametic disequilibria....

  9. Efficient production of Arthromyces ramosus peroxidase by Aspergillus awamori

    NARCIS (Netherlands)

    Lokman, B.C.; Joosten, V.; Hovenkamp, J.; Gouka, R.J.; Verrips, C.T.; Hondel, C.A.M.J.J. van den

    2003-01-01

    The heterologous production of Arthromyces ramosus peroxidase (ARP) was analysed in the filamentous fungus Aspergillus awamori under control of the inducible endoxylanase promoter. Secretion of active ARP was achieved up to 800 mg l-1 in shake flask cultures. Western blot analysis showed that an rAR

  10. KINETICS OF QUERCETIN NITRATIO N BY HORSERADISH PEROXIDASE

    Directory of Open Access Journals (Sweden)

    Andrija Šmelcerović

    2013-03-01

    Full Text Available In this study we investigated the kinetics of the nitration of quercetin by horseradish peroxidase. Quercetin nitration reaction was followed by recording the spectral changes over the time at 380 nm. The reaction rate increases with increasing of the quercetin concentration and follows the Michaelis-Menten type kinetics. Kinetic parameters of the studied enzymatic reaction were determined.

  11. An insight into the lignin peroxidase of Macrophomina phaseolina.

    Science.gov (United States)

    Akbar, Mohammed Touaha; Habib, Abdul Musaweer; Chowdhury, Dil Umme Salma; Bhuiyan, Md Iqbal Kaiser; Mostafa, Kazi Md Golam; Mondol, Sobuj; Mosleh, Ivan Mhai

    2013-01-01

    Macrophomina phaseolina is one of the deadliest necrotrophic fungal pathogens that infect more than 500 plant species including major food, fiber, and oil crops all throughout the globe. It secretes a cocktail of ligninolytic enzymes along with other hydrolytic enzymes for degrading the woody lignocellulosic plant cell wall and penetrating into the host tissue. Among them, lignin peroxidase has been reported only in Phanerochaete chrysosporium so far. But interestingly, a recent study has revealed a second occurrence of lignin peroxidase in M. phaseolina. However, lignin peroxidases are of much significance biotechnologically because of their potential applications in bio-remedial waste treatment and in catalyzing difficult chemical transformations. Besides, this enzyme also possesses agricultural and environmental importance on account of their role in lignin biodegradation. In the present work, different properties of the lignin peroxidase of M. phaseolina along with predicting the 3-D structure and its active sites were investigated by the use of various computational tools. The data from this study will pave the way for more detailed exploration of this enzyme in wet lab and thereby facilitating the strategies to be designed against such deadly weapons of Macrophomina phaseolina. Furthermore, the insight of such a ligninolytic enzyme will contribute to the assessment of its potentiality as a bioremediation tool.

  12. Efficient production of Arthromyces ramosus peroxidase by Aspergillus awamori

    NARCIS (Netherlands)

    Lokman, B.C.; Joosten, V.; Hovenkamp, J.; Gouka, R.J.; Verrips, C.T.; Hondel, C.A.M.J.J. van den

    2003-01-01

    The heterologous production of Arthromyces ramosus peroxidase (ARP) was analysed in the filamentous fungus Aspergillus awamori under control of the inducible endoxylanase promoter. Secretion of active ARP was achieved up to 800 mg l-1 in shake flask cultures. Western blot analysis showed that an

  13. Mammalian heme peroxidases: from molecular mechanisms to health implications.

    Science.gov (United States)

    Davies, Michael J; Hawkins, Clare L; Pattison, David I; Rees, Martin D

    2008-07-01

    A marked increase in interest has occurred over the last few years in the role that mammalian heme peroxidase enzymes, primarily myeloperoxidase, eosinophil peroxidase, and lactoperoxidase, may play in both disease prevention and human pathologies. This increased interest has been sparked by developments in our understanding of polymorphisms that control the levels of these enzymes, a greater understanding of the basic chemistry and biochemistry of the oxidants formed by these species, the development of specific biomarkers that can be used in vivo to detect damage induced by these oxidants, the detection of active forms of these peroxidases at most, if not all, sites of inflammation, and a correlation between the levels of these enzymes and a number of major human pathologies. This article reviews recent developments in our understanding of the enzymology, chemistry, biochemistry and biologic roles of mammalian peroxidases and the oxidants that they generate, the potential role of these oxidants in human disease, and the use of the levels of these enzymes in disease prognosis.

  14. The glucose oxidase-peroxidase assay for glucose

    Science.gov (United States)

    The glucose oxidase-peroxidase assay for glucose has served as a very specific, sensitive, and repeatable assay for detection of glucose in biological samples. It has been used successfully for analysis of glucose in samples from blood and urine, to analysis of glucose released from starch or glycog...

  15. Mechanism of reaction of chlorite with mammalian heme peroxidases.

    Science.gov (United States)

    Jakopitsch, Christa; Pirker, Katharina F; Flemmig, Jörg; Hofbauer, Stefan; Schlorke, Denise; Furtmüller, Paul G; Arnhold, Jürgen; Obinger, Christian

    2014-06-01

    This study demonstrates that heme peroxidases from different superfamilies react differently with chlorite. In contrast to plant peroxidases, like horseradish peroxidase (HRP), the mammalian counterparts myeloperoxidase (MPO) and lactoperoxidase (LPO) are rapidly and irreversibly inactivated by chlorite in the micromolar concentration range. Chlorite acts as efficient one-electron donor for Compound I and Compound II of MPO and LPO and reacts with the corresponding ferric resting states in a biphasic manner. The first (rapid) phase is shown to correspond to the formation of a MPO-chlorite high-spin complex, whereas during the second (slower) phase degradation of the prosthetic group was observed. Cyanide, chloride and hydrogen peroxide can block or delay heme bleaching. In contrast to HRP, the MPO/chlorite system does not mediate chlorination of target molecules. Irreversible inactivation is shown to include heme degradation, iron release and decrease in thermal stability. Differences between mammalian peroxidases and HRP are discussed with respect to differences in active site architecture and heme modification.

  16. Structural and spectroscopic characterisation of a heme peroxidase from sorghum.

    Science.gov (United States)

    Nnamchi, Chukwudi I; Parkin, Gary; Efimov, Igor; Basran, Jaswir; Kwon, Hanna; Svistunenko, Dimitri A; Agirre, Jon; Okolo, Bartholomew N; Moneke, Anene; Nwanguma, Bennett C; Moody, Peter C E; Raven, Emma L

    2016-03-01

    A cationic class III peroxidase from Sorghum bicolor was purified to homogeneity. The enzyme contains a high-spin heme, as evidenced by UV-visible spectroscopy and EPR. Steady state oxidation of guaiacol was demonstrated and the enzyme was shown to have higher activity in the presence of calcium ions. A Fe(III)/Fe(II) reduction potential of -266 mV vs NHE was determined. Stopped-flow experiments with H2O2 showed formation of a typical peroxidase Compound I species, which converts to Compound II in the presence of calcium. A crystal structure of the enzyme is reported, the first for a sorghum peroxidase. The structure reveals an active site that is analogous to those for other class I heme peroxidase, and a substrate binding site (assigned as arising from binding of indole-3-acetic acid) at the γ-heme edge. Metal binding sites are observed in the structure on the distal (assigned as a Na(+) ion) and proximal (assigned as a Ca(2+)) sides of the heme, which is consistent with the Ca(2+)-dependence of the steady state and pre-steady state kinetics. It is probably the case that the structural integrity (and, thus, the catalytic activity) of the sorghum enzyme is dependent on metal ion incorporation at these positions.

  17. Electrochemical Detection of Horseradish Peroxidase at Zeptomole Level

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    An electrochemical method for determination of horseradish peroxidase (HRP) was developed using a capillary catalytic system. HRP can be measured in several minutes with a detection limit of 4.8×10-12 mol/L or 47 zmol (S/N=3).

  18. Removal and isolation of germ-rich fractions from hull-less barley using a fitzpatrick comminuting mill

    Science.gov (United States)

    A process was developed to produce a germ-enriched fraction from hull-less barley using a Fitzpatrick Comminuting Mill followed by sieving. Hulled and hull-less barleys contain 1.5-2.5% oil and, like wheat kernels which contain wheat germ oil, much of the oil in barley kernels is in the germ fracti...

  19. Polimorfismo isozimático e potencial de utilização das isozimas como marcadores genéticos em aceroleira Isozyme polymorphism and potential of utilization of isozymes as genetics markers in West Indian cherry

    Directory of Open Access Journals (Sweden)

    Ricardo Lopes

    2002-02-01

    Full Text Available A aceroleira (Malpighia emarginata DC. é uma cultura em expansão no Brasil, principalmente, por causa do elevado teor de vitamina C de seus frutos; contudo, ainda são escassas as informações sobre a espécie. Este trabalho teve o objetivo de estudar a atividade e o polimorfismo de alguns sistemas isozimáticos de uma coleção de aceroleiras. Foram estudadas as isozimas IDH, MDH, EST, ACP, GOT, PGM, PGI e POD. Utilizou-se o sistema de eletroforese horizontal em gel de amido e diferentes tampões de gel e cuba. Identificou-se o número de locos e alelos envolvidos no controle genético das enzimas. IDH demonstrou ser dimérica, e foi constatado um loco e dois alelos. PGM apresentou dois locos diméricos, com dois e quatro alelos. EST e POD demonstraram ser monoméricas, com um e dois locos, respectivamente, todos com dois alelos. MDH apresentou um loco onde o seu comportamento é monomérico e outro dimérico, ambos com dois alelos. GOT foi polimórfico mas não foi possível inferir sobre seu controle genético. O polimorfismo revelado pelos sistemas IDH, MDH, POD, EST, GOT e PGM demonstra o potencial de utilização das isozimas como marcadores genéticos em estudos de conservação e melhoramento de aceroleira.The West Indian Cherry (Malpighia emarginata DC. is a crop in expansion in Brazil mainly due to high fruit vitamin C content. However, there is still little information about the species. The objective of this study was to determine the activity and the polymorphism of isozymes from a collection of West Indian Cherry plants. IDH, MDH, EST, POD, ACP, GOT, PGM and PGI isozymes were studied. Starch gel horizontal electrophoresis and different gel/electrode buffers were used. Number of loci and alleles involved in genetic control of these isozymes were identified. IDH was dimeric and had one locus with two alleles. PGM was monomeric and had two loci, with two and four alleles. POD and EST were monomeric and had one and two loci

  20. Degradation of textile dyes mediated by plant peroxidases.

    Science.gov (United States)

    Shaffiqu, T S; Roy, J Jegan; Nair, R Aswathi; Abraham, T Emilia

    2002-01-01

    The peroxidase enzyme from the plants Ipomea palmata (1.003 IU/g of leaf) and Saccharum spontaneum (3.6 IU/g of leaf) can be used as an alternative to the commercial source of horseradish and soybean peroxidase enzyme for the decolorization of textile dyes, mainly azo dyes. Eight textiles dyes currently used by the industry and seven other dyes were selected for decolorization studies at 25-200 mg/L levels using these plant enzymes. The enzymes were purified prior to use by ammonium sulfate precipitation, and ion exchange and gel permeation chromatographic techniques. Peroxidase of S. spontaneum leaf (specific activity of 0.23 IU/mg) could completely degrade Supranol Green and Procion Green HE-4BD (100%) dyes within 1 h, whereas Direct Blue, Procion Brilliant Blue H-7G and Chrysoidine were degraded >70% in 1 h. Peroxidase of Ipomea (I. palmata leaf; specific activity of 0.827 U/mg) degraded 50 mg/L of the dyes Methyl Orange (26%), Crystal Violet (36%), and Supranol Green (68%) in 2-4 h and Brilliant Green (54%), Direct Blue (15%), and Chrysoidine (44%) at the 25 mg/L level in 1 to 2 h of treatment. The Saccharum peroxidase was immobilized on a hydrophobic matrix. Four textile dyes, Procion Navy Blue HER, Procion Brilliant Blue H-7G, Procion Green HE-4BD, and Supranol Green, at an initial concentration of 50 mg/L were completely degraded within 8 h by the enzyme immobilized on the modified polyethylene matrix. The immobilized enzyme was used in a batch reactor for the degradation of Procion Green HE-4BD and the reusability was studied for 15 cycles, and the half-life was found to be 60 h.