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Sample records for isolated pea nuclei

  1. Characterization of nucleoside triphosphatase activity in isolated pea nuclei and its photoreversible regulation by light

    NASA Technical Reports Server (NTRS)

    Chen, Y. R.; Roux, S. J.

    1986-01-01

    A nucleoside triphosphatase (NTPase) present in highly purified preparations of pea nuclei was partially characterized. The activity of this enzyme was stimulated by divalent cations (Mg2+ = Mn2+ > Ca2+), but was not affected by the monovalent cations, Na+ and K+. The Mg(2+)-dependent activity was further stimulated by concentrations of Ca2+ in the low micromolar range. It could catalyze the hydrolysis of ATP, GTP, UTP, and CTP, all with a pH optimum of 7.5. The nuclear NTPase activity was not inhibited by vanadate, oligomycin, or nitrate, but was inhibited by relatively low concentrations of quercetin and the calmodulin inhibitor, compound 48/80. The NTPase was stimulated more than 50% by red light, and this effect was reversed by subsequent irradiation with far-red light. The photoreversibility of the stimulation indicated that the photoreceptor for this response was phytochrome, an important regulator of photomorphogenesis and gene expression in plants.

  2. Characterization of nucleoside triphosphatase activity in isolated pea nuclei and its photoreversible regulation by light

    NASA Technical Reports Server (NTRS)

    Chen, Y. R.; Roux, S. J.

    1986-01-01

    A nucleoside triphosphatase (NTPase) present in highly purified preparations of pea nuclei was partially characterized. The activity of this enzyme was stimulated by divalent cations (Mg2+ = Mn2+ > Ca2+), but was not affected by the monovalent cations, Na+ and K+. The Mg(2+)-dependent activity was further stimulated by concentrations of Ca2+ in the low micromolar range. It could catalyze the hydrolysis of ATP, GTP, UTP, and CTP, all with a pH optimum of 7.5. The nuclear NTPase activity was not inhibited by vanadate, oligomycin, or nitrate, but was inhibited by relatively low concentrations of quercetin and the calmodulin inhibitor, compound 48/80. The NTPase was stimulated more than 50% by red light, and this effect was reversed by subsequent irradiation with far-red light. The photoreversibility of the stimulation indicated that the photoreceptor for this response was phytochrome, an important regulator of photomorphogenesis and gene expression in plants.

  3. INCORPORATION OF TRITIATED CYTIDINE INTO RIBONUCLEIC ACID BY ISOLATED PEA NUCLEI

    PubMed Central

    Rho, Joon H.; Chipchase, Margaret I.

    1962-01-01

    A new method for the isolation of the nuclei from plant tissue has been devised. This method consists in passing the plant tissue through a set of spring-loaded, counter-rotating rollers and collecting the liberated nuclei in sucrose solution containing calcium ions. A semi-automatic machine which couples the rollers with a special tissue chopping device has been constructed. It has been shown that nuclei obtained in this way actively incorporate cytidine-H3 into RNA. This incorporation is increased in the presence of nucleoside triphosphates and an energy-regenerating system. PMID:14491776

  4. Regulation of the transcription of storage-protein mRNA in nuclei isolated from developing pea (Pisum sativum L.) cotyledons.

    PubMed

    Evans, I M; Gatehouse, J A; Croy, R R; Boulter, D

    1984-05-01

    Two types of storage protein, vicillin and legumin, occur in the developing pea seed. Storage-protein gene expression has been studied during cotyledon development by assaying specific transcripts produced by nuclei isolated at different stages, and from pea leaves. The proportion of vicilin to legumin transcripts changed during development: vicilin transcripts predominated at 9 and 11 days after flower opening (d.a.f.) and were similar in amount to legumin at 14 d.a.f., whereas at 18 d.a.f., legumin transcripts predominated and little vicilin transcription was observed. The rate of storage-protein transcription correlated with previously determined (Gatehouse et al. 1982) mRNA levels during seed development; these transcripts were not detected in similar assays using leaf nuclei. Transcription by cotyledonary nuclei for short times indicated that post-transcriptional processing may be a factor in regulating mRNA levels, at least in the earlier part of seed development.

  5. Casein kinase II protein kinase is bound to lamina-matrix and phosphorylates lamin-like protein in isolated pea nuclei

    NASA Technical Reports Server (NTRS)

    Li, H.; Roux, S. J.

    1992-01-01

    A casein kinase II (CK II)-like protein kinase was identified and partially isolated from a purified envelope-matrix fraction of pea (Pisum sativum L.) nuclei. When [gamma-32P]ATP was directly added to the envelope-matrix preparation, the three most heavily labeled protein bands had molecular masses near 71, 48, and 46 kDa. Protein kinases were removed from the preparation by sequential extraction with Triton X-100, EGTA, 0.3 M NaCl, and a pH 10.5 buffer, but an active kinase still remained bound to the remaining lamina-matrix fraction after these treatments. This kinase had properties resembling CK II kinases previously characterized from animal and plant sources: it preferred casein as an artificial substrate, could use GTP as efficiently as ATP as the phosphoryl donor, was stimulated by spermine, was calcium independent, and had a catalytic subunit of 36 kDa. Some animal and plant CK II kinases have regulatory subunits near 29 kDa, and a lamina-matrix-bound protein of this molecular mass was recognized on immunoblot by anti-Drosophila CK II polyclonal antibodies. Also found associated with the envelope-matrix fraction of pea nuclei were p34cdc2-like and Ca(2+)-dependent protein kinases, but their properties could not account for the protein kinase activity bound to the lamina. The 71-kDa substrate of the CK II-like kinase was lamin A-like, both in its molecular mass and in its cross-reactivity with anti-intermediate filament antibodies. Lamin phosphorylation is considered a crucial early step in the entry of cells into mitosis, so lamina-bound CK II kinases may be important control points for cellular proliferation.

  6. Casein kinase II protein kinase is bound to lamina-matrix and phosphorylates lamin-like protein in isolated pea nuclei

    NASA Technical Reports Server (NTRS)

    Li, H.; Roux, S. J.

    1992-01-01

    A casein kinase II (CK II)-like protein kinase was identified and partially isolated from a purified envelope-matrix fraction of pea (Pisum sativum L.) nuclei. When [gamma-32P]ATP was directly added to the envelope-matrix preparation, the three most heavily labeled protein bands had molecular masses near 71, 48, and 46 kDa. Protein kinases were removed from the preparation by sequential extraction with Triton X-100, EGTA, 0.3 M NaCl, and a pH 10.5 buffer, but an active kinase still remained bound to the remaining lamina-matrix fraction after these treatments. This kinase had properties resembling CK II kinases previously characterized from animal and plant sources: it preferred casein as an artificial substrate, could use GTP as efficiently as ATP as the phosphoryl donor, was stimulated by spermine, was calcium independent, and had a catalytic subunit of 36 kDa. Some animal and plant CK II kinases have regulatory subunits near 29 kDa, and a lamina-matrix-bound protein of this molecular mass was recognized on immunoblot by anti-Drosophila CK II polyclonal antibodies. Also found associated with the envelope-matrix fraction of pea nuclei were p34cdc2-like and Ca(2+)-dependent protein kinases, but their properties could not account for the protein kinase activity bound to the lamina. The 71-kDa substrate of the CK II-like kinase was lamin A-like, both in its molecular mass and in its cross-reactivity with anti-intermediate filament antibodies. Lamin phosphorylation is considered a crucial early step in the entry of cells into mitosis, so lamina-bound CK II kinases may be important control points for cellular proliferation.

  7. Isolation of Nuclei.

    PubMed

    Nabbi, Arash; Riabowol, Karl

    2015-08-03

    The isolation of nuclei is often the first step in studying processes such as nuclear-cytoplasmic shuttling, subcellular localization of proteins, and protein-chromatin or nuclear protein-protein interactions in response to diverse stimuli. Therefore, rapidly obtaining nuclei from cells with relatively high purity and minimal subcellular contamination, protein degradation, or postharvesting modification is highly desirable. Historically, the isolation of nuclei involved a homogenization step followed by centrifugation through high-density glycerol or sucrose. Although clean nuclei with little cytoplasmic contamination can be prepared using this method, it is typically time consuming and can allow protein degradation, protein modification, and leaching of components from the nuclei to occur. We have developed a rapid and simple fractionation method that is based on the selective dissolution of the cytoplasmic membrane (but not the nuclear membrane) using a low concentration of a nonionic detergent and rapid centrifugation steps. Here we describe important considerations when isolating nuclei from cells, introduce our rapid method, and compare this method to a more traditional protocol for isolating nuclei, noting the strengths and limitations of each approach.

  8. Phytochrome regulates GTP-binding protein activity in the envelope of pea nuclei

    NASA Technical Reports Server (NTRS)

    Clark, G. B.; Memon, A. R.; Thompson, G. A. Jr; Roux, S. J.

    1993-01-01

    Three GTP-binding proteins with apparent molecular masses of 27, 28 and 30 kDa have been detected in isolated nuclei of etiolated pea plumules. After LDS-PAGE and transfer to nitrocellulose these proteins bind [32P]GTP in the presence of excess ATP, suggesting that they are monomeric G proteins. When nuclei are disrupted, three proteins co-purify with the nuclear envelope fraction and are highly enriched in this fraction. The level of [32P]GTP-binding for all three protein bands is significantly increased when harvested pea plumules are irradiated by red light, and this effect is reversed by far-red light. The results indicate that GTP-binding activity associated with the nuclear envelope of plant cells is photoreversibly regulated by the pigment phytochrome.

  9. Phytochrome regulates GTP-binding protein activity in the envelope of pea nuclei

    NASA Technical Reports Server (NTRS)

    Clark, G. B.; Memon, A. R.; Thompson, G. A. Jr; Roux, S. J.

    1993-01-01

    Three GTP-binding proteins with apparent molecular masses of 27, 28 and 30 kDa have been detected in isolated nuclei of etiolated pea plumules. After LDS-PAGE and transfer to nitrocellulose these proteins bind [32P]GTP in the presence of excess ATP, suggesting that they are monomeric G proteins. When nuclei are disrupted, three proteins co-purify with the nuclear envelope fraction and are highly enriched in this fraction. The level of [32P]GTP-binding for all three protein bands is significantly increased when harvested pea plumules are irradiated by red light, and this effect is reversed by far-red light. The results indicate that GTP-binding activity associated with the nuclear envelope of plant cells is photoreversibly regulated by the pigment phytochrome.

  10. Partial purification and characterization of a Ca(2+)-dependent protein kinase from pea nuclei

    NASA Technical Reports Server (NTRS)

    Li, H.; Dauwalder, M.; Roux, S. J.

    1991-01-01

    Almost all the Ca(2+)-dependent protein kinase activity in nuclei purified from etiolated pea (Pisum sativum, L.) plumules is present in a single enzyme that can be extracted from chromatin by 0.3 molar NaCl. This protein kinase can be further purified 80,000-fold by salt fractionation and high performance liquid chromatography, after which it has a high specific activity of about 100 picomoles per minute per microgram in the presence of Ca2+ and reaches half-maximal activation at about 3 x 10(-7) molar free Ca2+, without calmodulin. It is a monomer with a molecular weight near 90,000. It can efficiently use histone III-S, ribosomal S6 protein, and casein as artificial substrates, but it phosphorylates phosvitin only weakly. Its Ca(2+)-dependent kinase activity is half-maximally inhibited by 0.1 millimolar chlorpromazine, by 35 nanomolar K-252a and by 7 nanomolar staurosporine. It is insensitive to sphingosine, an inhibitor of protein kinase C, and to basic polypeptides that block other Ca(2+)-dependent protein kinases. It is not stimulated by exogenous phospholipids or fatty acids. In intact isolated pea nuclei it preferentially phosphorylates several chromatin-associated proteins, with the most phosphorylated protein band being near the same molecular weight (43,000) as a nuclear protein substrate whose phosphorylation has been reported to be stimulated by phytochrome in a calcium-dependent fashion.

  11. Partial purification and characterization of a Ca(2+)-dependent protein kinase from pea nuclei

    NASA Technical Reports Server (NTRS)

    Li, H.; Dauwalder, M.; Roux, S. J.

    1991-01-01

    Almost all the Ca(2+)-dependent protein kinase activity in nuclei purified from etiolated pea (Pisum sativum, L.) plumules is present in a single enzyme that can be extracted from chromatin by 0.3 molar NaCl. This protein kinase can be further purified 80,000-fold by salt fractionation and high performance liquid chromatography, after which it has a high specific activity of about 100 picomoles per minute per microgram in the presence of Ca2+ and reaches half-maximal activation at about 3 x 10(-7) molar free Ca2+, without calmodulin. It is a monomer with a molecular weight near 90,000. It can efficiently use histone III-S, ribosomal S6 protein, and casein as artificial substrates, but it phosphorylates phosvitin only weakly. Its Ca(2+)-dependent kinase activity is half-maximally inhibited by 0.1 millimolar chlorpromazine, by 35 nanomolar K-252a and by 7 nanomolar staurosporine. It is insensitive to sphingosine, an inhibitor of protein kinase C, and to basic polypeptides that block other Ca(2+)-dependent protein kinases. It is not stimulated by exogenous phospholipids or fatty acids. In intact isolated pea nuclei it preferentially phosphorylates several chromatin-associated proteins, with the most phosphorylated protein band being near the same molecular weight (43,000) as a nuclear protein substrate whose phosphorylation has been reported to be stimulated by phytochrome in a calcium-dependent fashion.

  12. Glycerolipid biosynthesis in isolated pea root plastids

    SciTech Connect

    Xue, Lingru; Sparace, S.A. )

    1990-05-01

    Plastids have been isolated from germinating pea (Pisum sativum L.) roots by differential centrifugation and purified on Percoll gradients. Marker enzymes (NADPH: cytochrome c reductase, fumarase and fatty acid synthesis) indicate that greater than 50% of the plastids are recovered essentially free from mitochondrial and endoplasmic reticulum contamination. Fatty acids synthesized from ({sup 14}C)acetate by Percoll-purified plastids are primarily 16:0, 16:1 and 18:1. ({sup 14}C)Acetate-labelled fatty acids and ({sup 14}C)glycerol-3-phosphate are both readily incorporated into glycerolipid. Approximately 12% of the total activity for glycerolipid biosynthesis from glycerol-3-phosphate is recovered in the purified plastid fraction. Glycerolipids synthesized from these precursors are primarily TAG, DAG, PE, PG, PC, PI and PA. Acyl-CoA's also accumulate when acetate is the precursor.

  13. Protein import into isolated pea root leucoplasts

    PubMed Central

    Chu, Chiung-Chih; Li, Hsou-min

    2015-01-01

    Leucoplasts are important organelles for the synthesis and storage of starch, lipids and proteins. However, molecular mechanism of protein import into leucoplasts and how it differs from that of import into chloroplasts remain unknown. We used pea seedlings for both chloroplast and leucoplast isolations to compare within the same species. We further optimized the isolation and import conditions to improve import efficiency and to permit a quantitative comparison between the two plastid types. The authenticity of the import was verified using a mitochondrial precursor protein. Our results show that, when normalized to Toc75, most translocon proteins are less abundant in leucoplasts than in chloroplasts. A precursor shown to prefer the receptor Toc132 indeed had relatively more similar import efficiencies between chloroplasts and leucoplasts compared to precursors that prefer Toc159. Furthermore we found two precursors that exhibited very high import efficiency into leucoplasts. Their transit peptides may be candidates for delivering transgenic proteins into leucoplasts and for analyzing motifs important for leucoplast import. PMID:26388889

  14. Isolation and forward genetic analysis of developmental genes in pea.

    PubMed

    Weller, James L; Hecht, Valérie F G; Sussmilch, Frances C

    2013-01-01

    Understanding of developmental processes relies heavily on isolation and functional characterization of relevant genes. The garden pea (Pisum sativum L.) is one of the classic model species in plant genetics and has been used for a wide range of physiological and molecular studies of plant development. Here we describe the resources and approaches available for isolation of genes and genetic characterization of loci affecting development in pea.

  15. The major nucleoside triphosphatase in pea (Pisum sativum L.) nuclei and in rat liver nuclei share common epitopes also present in nuclear lamins

    NASA Technical Reports Server (NTRS)

    Tong, C. G.; Dauwalder, M.; Clawson, G. A.; Hatem, C. L.; Roux, S. J.

    1993-01-01

    The major nucleoside triphosphatase (NTPase) activities in mammalian and pea (Pisum sativum L.) nuclei are associated with enzymes that are very similar both biochemically and immunochemically. The major NTPase from rat liver nuclei appears to be a 46-kD enzyme that represents the N-terminal portion of lamins A and C, two lamina proteins that apparently arise from the same gene by alternate splicing. Monoclonal antibody (MAb) G2, raised to human lamin C, both immunoprecipitates the major (47 kD) NTPase in pea nuclei and recognizes it in western blot analyses. A polyclonal antibody preparation raised to the 47-kD pea NTPase (pc480) reacts with the same lamin bands that are recognized by MAb G2 in mammalian nuclei. The pc480 antibodies also bind to the same lamin-like bands in pea nuclear envelope-matrix preparations that are recognized by G2 and three other MAbs known to bind to mammalian lamins. In immunofluorescence assays, pc480 and anti-lamin antibodies stain both cytoplasmic and nuclear antigens in plant cells, with slightly enhanced staining along the periphery of the nuclei. These results indicate that the pea and rat liver NTPases are structurally similar and that, in pea nuclei as in rat liver nuclei, the major NTPase is probably derived from a lamin precursor by proteolysis.

  16. The major nucleoside triphosphatase in pea (Pisum sativum L.) nuclei and in rat liver nuclei share common epitopes also present in nuclear lamins

    NASA Technical Reports Server (NTRS)

    Tong, C. G.; Dauwalder, M.; Clawson, G. A.; Hatem, C. L.; Roux, S. J.

    1993-01-01

    The major nucleoside triphosphatase (NTPase) activities in mammalian and pea (Pisum sativum L.) nuclei are associated with enzymes that are very similar both biochemically and immunochemically. The major NTPase from rat liver nuclei appears to be a 46-kD enzyme that represents the N-terminal portion of lamins A and C, two lamina proteins that apparently arise from the same gene by alternate splicing. Monoclonal antibody (MAb) G2, raised to human lamin C, both immunoprecipitates the major (47 kD) NTPase in pea nuclei and recognizes it in western blot analyses. A polyclonal antibody preparation raised to the 47-kD pea NTPase (pc480) reacts with the same lamin bands that are recognized by MAb G2 in mammalian nuclei. The pc480 antibodies also bind to the same lamin-like bands in pea nuclear envelope-matrix preparations that are recognized by G2 and three other MAbs known to bind to mammalian lamins. In immunofluorescence assays, pc480 and anti-lamin antibodies stain both cytoplasmic and nuclear antigens in plant cells, with slightly enhanced staining along the periphery of the nuclei. These results indicate that the pea and rat liver NTPases are structurally similar and that, in pea nuclei as in rat liver nuclei, the major NTPase is probably derived from a lamin precursor by proteolysis.

  17. Transcription in Isolated Wheat Nuclei

    PubMed Central

    Luthe, Dawn Sywassink; Quatrano, Ralph S.

    1980-01-01

    Nuclei free of RNase activity were isolated from 3-hour-imbibed wheat (var. Yamhill) embryos by centrifugation through a discontinuous gradient of Percoll. The maximum rate of RNA synthesis observed in these nuclei was approximately 5 picomoles [3H]UTP per milligram DNA per minute. Two monovalent cation optima were found when measured in the presence of 15 millimolar MgCl2 or 2 millimolar MnCl2; 15 and 75 millimolar (NH4)2SO4. At the high monovalent cation optimum, the rate of RNA synthesis was linear for the first 10 to 15 minutes of incubation and still increasing after 3 hours. RNA synthesized in vitro (30-minute pulse followed by a 30-minute chase) showed distinct 18 and 26S RNA peaks comprising 13 and 17% of the total radioactivity, respectively. The over-all pattern of RNA synthesized in vitro was similar to the in vivo pattern. Approximately 40 to 50% of the RNA synthesized was inhibited by α-amanitin at 4 micrograms per milliliter. The newly synthesized 6 to 10S RNA appeared to be selectively inhibited by α-amanitin. PMID:16661179

  18. ISOLATION OF SKELETAL MUSCLE NUCLEI

    PubMed Central

    Edelman, Jean C.; Edelman, P. Michael; Knigge, Karl M.; Schwartz, Irving L.

    1965-01-01

    A method employing aqueous media for isolation of nuclei from rat skeletal muscle is described. The technique involves (a) mincing and then homogenizing in a 0.32 M sucrose-salt solution with a Potter-Elvehjem type homogenizer using a Delrin (an acetal resin) pestle and a carefully controlled, relatively large pestle-to-glass clearance, (b) filtering through fiberglass and stainless steel screens of predetermined mesh size to remove myofibrils and connective tissue, and (c) centrifuging in a 2.15 M sucrose-salt solution containing 0.7 mM ATP. Electron and phase-contrast microscopic observations show that the nuclei are intact, unencumbered by cytoplasmic tags, and possess well preserved distinct nucleoli, nucleoplasm, and nuclear membranes. Cytoplasmic contamination is minimal and mainly mitochondrial. Chemical assays of the nuclear fraction show that the DNA/protein and RNA/DNA ratios are comparable to those obtained in other tissues. These ratios, as well as the low specific activity obtained for cytochrome c oxidase and the virtual absence of myofibrillar ATPase, indicate a high degree of purity with minimal mitochondrial and myofibrillar contamination. The steps comprising the technique and the reasons for their selection are discussed. PMID:4287141

  19. Isolation and study of the functional properties of pea proteins.

    PubMed

    Tömösközi, S; Lásztity, R; Haraszi, R; Baticz, O

    2001-10-01

    Proteins of pea seeds were isolated after defatting with hexane using alkaline (0.1 M sodium hydroxide) extraction and acid (HCl) precipitation. Concentrates were also prepared by hexane extraction and ethanolic extraction (pH = 5). Gross chemical composition amino acid content and functional properties (solubility profile, emulsifying--and foaming properties, water--and oil absorption) were studied. The results were compared with the same parameters of soy and lupin protein products. Although the majority of functional characteristics of isolates were lower in comparison to soy isolates, pea protein concentrate and isolate could be successfully used in bakery products for enrichment in protein and improvement of biological value. Their utilization as meat protein substitute in some Frankfurter type sausages is also possibly.

  20. PHYSICAL STUDIES OF ISOLATED EUCARYOTIC NUCLEI

    PubMed Central

    Olins, Donald E.; Olins, Ada L.

    1972-01-01

    The degree of chromatin condensation in isolated rat liver nuclei and chicken erythrocyte nuclei was studied by phase-contrast microscopy as a function of solvent pH, K+ and Mg++ concentrations Data were represented as "phase" maps, and standard solvent conditions selected that reproducibly yield granular, slightly granular, and homogeneous nuclei Nuclei in these various states were examined by ultraviolet absorption and circular dichroism (CD) spectroscopy, low-angle X-ray diffraction, electron microscopy, and binding capacity for ethidium bromide Homogeneous nuclei exhibited absorption and CD spectra resembling those of isolated nucleohistone. Suspensions of granular nuclei showed marked turbidity and absorption flattening, and a characteristic blue-shift of a crossover wavelength in the CD spectra. In all solvent conditions studied, except pH < 2 3, low-angle X-ray reflections characteristic of the native, presumably superhelical, nucleohistone were observed from pellets of intact nuclei. Threads (100–200 A diameter) were present in the condensed and dispersed phases of nuclei fixed under the standard solvent conditions, and examined in the electron microscope after thin sectioning and staining Nuclei at neutral pH, with different degrees of chromatin condensation, exhibited similar binding capacities for ethidium bromide. These data suggest a model that views chromatin condensation as a close packing of superhelical nucleohistone threads but still permits condensed chromatin to respond rapidly to alterations in solvent environment. PMID:4554987

  1. CN(-)-Induced degradation of nuclei in cells of pea leaves.

    PubMed

    Samuilov, V D; Lagunova, E M; Beshta, O E; Kitashov, A V

    2000-06-01

    Degradation of nuclei in epidermal and guard cells of pea leaves was induced by NaCN. Guard cells were considerably more resistant to CN- than epidermal cells. CN--induced nucleus degradation in guard cells was accelerated by illumination. The effect of illumination was negligible in epidermal cells that, unlike guard cells, do not contain chloroplasts. These data may indicate a role of chloroplasts in CN--induced cell death. CN--induced nucleus degradation in epidermal cells was retarded by antioxidants (butylated hydroxytoluene and vitamin E). The effect of CN- in guard cells was largely removed by vitamin E. Salicylic acid, an inhibitor of catalase and ascorbate peroxidase, induced 100% degradation of nuclei in epidermal cells but did not significantly affect nuclei in guard cells. CN--induced inhibition of catalase and peroxidase is assumed to lead to generation and accumulation of reactive oxygen species inducing apoptosis. Like mitochondria, which play an important role in animal cell apoptosis, chloroplasts may take part in apoptosis in plant cells.

  2. Purification and characterization of a casein kinase 2-type protein kinase from pea nuclei

    NASA Technical Reports Server (NTRS)

    Li, H.; Roux, S. J.

    1992-01-01

    Almost all the polyamine-stimulated protein kinase activity associated with the chromatin fraction of nuclei purified from etiolated pea (Pisum sativum L.) plumules is present in a single enzyme that can be extracted from chromatin by 0.35 molar NaCl. This protein kinase can be further purified over 2000-fold by salt fractionation and anion-exchange and casein-agarose column chromatography, after which it is more than 90% pure. The purified kinase has a specific activity of about 650 nanomoles per minute per milligram protein in the absence of polyamines, with either ATP or GTP as phosphoryl donor. Spermidine can stimulate its activity fourfold, with half-maximal activation at about 2 millimolar. Spermine and putrescine also stimulate activity, although somewhat less effectively. This kinase has a tetrameric alpha 2 beta 2 structure with a native molecular weight of 130,000, and subunit molecular weights of 36,000 for the catalytic subunit (alpha) and 29,000 for the regulatory subunit (beta). In western blot analyses, only the alpha subunit reacts strongly with polyclonal antibodies to a Drosophila casein kinase II. The pea kinase can use casein and phosvitin as artificial substrates, phosphorylating both the serine and threonine residues of casein. It has a pH optimum near 8.0, a Vmax of 1.5 micromoles per minute per milligram protein, and a Km for ATP of approximately 75 micromolar. Its activity can be almost completely inhibited by heparin at 5 micrograms per milliliter, but is relatively insensitive to concentrations of staurosporine, K252a, and chlorpromazine that strongly antagonize Ca(2+) -regulated protein kinases. These results are discussed in relation to recent findings that casein kinase 2-type kinases may phosphorylate trans-acting factors that bind to light-regulated promoters in plants.

  3. Pentraxin binding to isolated rat liver nuclei.

    PubMed Central

    Shephard, E G; Smith, P J; Coetzee, S; Strachan, A F; de Beer, F C

    1991-01-01

    The interaction of human C-reactive protein (CRP) and serum amyloid P-component (SAP) with isolated rat liver nuclei was studied to identify nuclear ligands for each pentraxin using the iodinatable heterobifunctional thiol-cleavable cross-linking reagent sulphosuccinimidyl-2-(p-azidosalicylamido)-1,3'-dithiopropio nate (SASD). Nuclei (100 micrograms of DNA) bound 21 pmol of 125I-labelled CRP Ca(2+)-dependently at saturation with half-saturation occurring at 200 pmol of 125I-CRP. By contrast, only 2.7 pmol of 125I-labelled SAP was bound at saturation, with half-saturation at 50 pmol. The binding of pentraxins to nuclei is, in addition to putative chromatin binding, due to nuclear-envelope binding, where 3.2 pmol 125I-labelled CRP binds Ca2+ dependently to nuclear envelopes (25 micrograms) at saturation, but only 0.62 pmol SAP is required to saturate. Specific photocross-linking of 125I-2-(p-azidosalicylamido)-1,3'-dithiopropionate (125I-ASD)-CRP and 125I-ASD-SAP to nuclei revealed transfer of 125I-photoreactive azides to nuclear-envelope proteins of 43, 46, 52 and 70 kDa. In addition, SAP binding to histones H2A, H2B, H3 and H4 was detected, whereas CRP bound only to H4. Neither pentraxin cross-linked to histone H1. Images Fig. 2. Fig. 3. Fig. 4. PMID:1930144

  4. DAPI fluorescence in nuclei isolated from tumors.

    PubMed

    Krishan, Awtar; Dandekar, Payal D

    2005-08-01

    In DNA histograms of some human solid tumors stained with nuclear isolation medium--4,6-diamidino-2-phenylindole dihydrochloride (NIM-DAPI), the coefficient of variation (CV) of the G0/G1 peak was broad, and in nuclear volume vs DNA scattergrams, a prominent slope was seen. To determine the cause for this, nuclei from frozen breast tumors were stained with NIM-DAPI and analyzed after dilution or resuspension in PBS. In two-color (blue vs red) analysis, most of the slope and broad CV was due to red fluorescence of nuclei stained with NIM-DAPI, which was reduced on dilution or resuspension in PBS, resulting in elimination of the slope and tightening of the CV.

  5. Formation of putative chloroplast cytochromes in isolated developing pea chloroplasts

    SciTech Connect

    Thaver, S.S.; Bhava, D.; Castelfranco, P.A.

    1986-04-01

    In addition to chlorophyll-protein complexes, other proteins were labeled when isolated developing pea chloroplasts were incubated with (/sup 14/C)-5-aminolevulinic acid (/sup 14/C)-ALA. The major labeled band (M/sub r/ = 43 kDa by LDS-PAGE) was labeled even in the presence of chloramphenicol. Heme-dependent peroxidase activity (as detected by the tetramethyl benzidine-H/sub 2/O/sub 2/ stain) was not visibly associated with this band. The radioactive band was stable to heat, 5% HCl in acetone, and was absent if the incubation with (/sup 14/C)-5-aminolevulinic acid was carried out in the presence of N-methyl protoporphyrin IX dimethyl ester (a specific inhibitor of ferrochelatase). Organic solvent extraction procedures for the enrichment of cytochrome f from chloroplast membranes also extracted this unknown labeled product. It was concluded that this labeled product was probably a c-type cytochrome. The effect of exogenous iron, iron chelators, gabaculine (an inhibitor of ALA synthesis) and other incubation conditions upon the in vitro formation of putative chloroplast cytochromes will be discussed.

  6. Comparative study of the functional properties of three legume seed isolates: adzuki, pea and soy bean.

    PubMed

    Barac, Miroljub B; Pesic, Mirjana B; Stanojevic, Sladjana P; Kostic, Aleksandar Z; Bivolarevic, Vanja

    2015-05-01

    The aim of this work was to compare functional properties including solubility, emulsifying and foaming properties of native and thermally treated adzuki, soy and pea protein isolates prepared under the same conditions. These functional properties were tested at four pH values: pH 3.0, pH 5.0, pH 7.0 and pH 8.0. The lowest solubility at all pH values were obtained for isolate of adzuki whereas isolates of soybean had the highest values at almost all pHs. Thermal treatment reduced solubility of soy and pea isolates at all pH values, whereas solubility of adzuki isolate was unchanged, except at pH 8. Native isolate of adzuki had the best emulsifying properties at pH 7.0 whereas at the other pH values some of native pea and soybean protein isolates were superior. After thermal treatment, depending on tested pH and selected variety all of three species could be a good emulsifier. Native soy protein isolates formed the most stable foams at all pHs. Thermal treatment significantly improved foaming properties of adzuki isolate, whereas reduced foaming capacity of soy and pea isolates, but could improve foam stability of these isolates at specific pH. Appropriate selection of legume seed as well as variety could have great importance in achievement of desirable functional properties of final products. All three tested species could find specific application in wide range of food products.

  7. Characterization of Five Fungal Endophytes Producing Cajaninstilbene Acid Isolated from Pigeon Pea [Cajanus cajan (L.) Millsp.

    PubMed Central

    Zu, Yuan Gang; Fu, Yu Jie; Wang, Wei; Luo, Meng; Efferth, Thomas

    2011-01-01

    Five fungal endophytes (K4, K5, K6, K9, K14) producing Cajaninstilbene acid (CSA, 3-hydroxy-4-prenyl-5-methoxystilbene-2-carboxylic acid) were isolated from the roots of pigeon pea [Cajanus cajan (L.) Millsp.]. CSA is responsible for the prominent pharmacological activities in pigeon pea. The amount of CSA in culture solution varied among the five fungal endophytes. K4 produced the highest levels of CSA (1037.13 µg/L) among the endophytes tested after incubation for five days. Both morphological characteristics and molecular methods were used for species identification of fungal endophytes. The five endophytic isolates were characterized by analyzing the internal transcribed spacer (ITS) rRNA and β-tubulin genes. The K4, K5, K9 and K14 strains isolated from pigeon pea roots were found to be closely related to the species Fusarium oxysporum. K6 was identified as Neonectria macrodidym. The present study is the first report on the isolation and identification of fungal endophytes producing CSA in pigeon pea. The study also provides a scientific base for large scale production of CSA. PMID:22102911

  8. Protein Synthesis by Isolated Etioplasts and Chloroplasts from Pea and Wheat and the Effects of Chloramphenicol and Cycloheximide 1

    PubMed Central

    Reger, Bonnie J.; Smillie, R. M.; Fuller, R. C.

    1972-01-01

    Etioplasts capable of incorporating 14C-leucine into protein have been isolated from dark-grown pea and wheat plants. The requirements for leucine incorporation for etioplasts were similar to those for chloroplasts. An ATP-generating system, Mg2+, and GTP were required. The amino-acid-incorporation activity of etioplasts from wheat was comparable to that of chloroplasts on an RNA basis, whereas the activity of pea etioplasts was about 50% of the activity of pea chloroplasts. The incorporation of leucine into protein by etioplasts and chloroplasts from pea and wheat was inhibited by chloramphenicol, and to a slight extent by cycloheximide. PMID:16658121

  9. Gibberellin metabolism in isolated pea fruit tissue and intact fruits

    SciTech Connect

    Maki, S.; Brenner, M.L. )

    1989-04-01

    Gibberellins (GAs) have been shown by others to be required for normal development of pea fruit. Whether the pericarp of the developing pea fruit produces GAs in situ is not known. To determine if the pericarp has the capacity to produce GAs during fruit growth, the metabolism of the first two committed GAs in the biosynthetic pathway, ({sup 14}C)GA{sub 12}-aldehyde and ({sup 14}C)GA{sub 12} was examined in tissue obtained from pollinated, parthenocarpic, and control fruit over 4 days from treatment. ({sup 14}C)GA{sub 12}-aldehyde was converted primarily to conjugates, including ({sup 14}C)GA{sub 12}-aldehyde conjugate. ({sup 14}C)GA{sub 12} was converted to ({sup 14}C)GA{sub 53} in all tissue, but by day 4 only tissue from pollinated or parthenocarpic fruits showed sustained formation of ({sup 14}C)GA{sub 53}. When ({sup 14}C)GA{sub 12} is applied to 4-day-old fruits attached to the plants, the major product obtained after 24 hours is ({sup 14}C)GA{sub 20} (as identified by GC-MS). No transport to the developing seed was observed. These results indicate that the elongating fruit tissue has the capacity to produce GAs.

  10. THE ISOLATION OF CELL NUCLEI IN NON-AQUEOUS MEDIA

    PubMed Central

    Allfrey, V.; Stern, H.; Mirsky, A. E.; Saetren, H.

    1952-01-01

    1. A modified Behrens procedure is described for the isolation of nuclei from avian erythrocytes and from the liver, kidney, thymus, pancreas, heart, and intestinal mucosa of the calf or horse. 2. The purity of these nuclei has been established by staining reactions, enzyme studies, and immunological tests for serum proteins. 3. Evidence is presented to show that a transport of cytoplasmic proteins into the nucleus does not occur during the isolation. 4. Nuclei prepared in non-aqueous media contain considerably more protein and a very different enzyme composition from that observed in nuclei prepared by "homogenization" techniques in dilute citric acid. 5. The suitability of nuclei prepared in organic media for the study of intracellular enzyme distribution is discussed. PMID:14898034

  11. Functional Properties of Pea (Pisum sativum, L.) Protein Isolates Modified with Chymosin

    PubMed Central

    Barać, Miroljub; Čabrilo, Slavica; Pešić, Mirjana; Stanojević, Slađana; Pavlićević, Milica; Maćej, Ognjen; Ristić, Nikola

    2011-01-01

    In this paper, the effects of limited hydrolysis on functional properties, as well as on protein composition of laboratory-prepared pea protein isolates, were investigated. Pea protein isolates were hydrolyzed for either 15, 30 and 60 min with recombined chymosin (Maxiren). The effect of enzymatic action on solubility, emulsifying and foaming properties at different pH values (3.0; 5.0; 7.0 and 8.0) was monitored. Chymosin can be a very useful agent for improvement of functional properties of isolates. Action of this enzyme caused a low degree of hydrolysis (3.9–4.7%), but improved significantly functional properties of pea protein isolates (PPI), especially at lower pH values (3.0–5.0). At these pH values all hydrolysates had better solubility, emulsifying activity and foaming stability, while longer-treated samples (60 min) formed more stable emulsions at higher pH values (7.0, 8.0) than initial isolates. Also, regardless of pH value, all hydrolysates showed improved foaming ability. A moderate positive correlation between solubility and emulsifying activity index (EAI) (0.74) and negative correlation between solubility and foam stability (−0.60) as well as between foam stability (FS) and EAI (−0.77) were observed. Detected enhancement in functional properties was a result of partial hydrolysis of insoluble protein complexes. PMID:22272078

  12. Isolation, selection, and characterization of beneficial rhizobacteria from pea, lentil, and chickpea grown in western Canada.

    PubMed

    Hynes, Russell K; Leung, Grant C Y; Hirkala, Danielle L M; Nelson, Louise M

    2008-04-01

    The use of beneficial soil microorganisms as agricultural inputs for improved crop production requires selection of rhizosphere-competent microorganisms with plant growth-promoting attributes. A collection of 563 bacteria originating from the roots of pea, lentil, and chickpea grown in Saskatchewan was screened for several plant growth-promoting traits, for suppression of legume fungal pathogens, and for plant growth promotion. Siderophore production was detected in 427 isolates (76%), amino-cyclopropane-1-carboxylic acid (ACC) deaminase activity in 29 isolates (5%), and indole production in 38 isolates (7%). Twenty-six isolates (5%) suppressed the growth of Pythium sp. strain p88-p3, 40 isolates (7%) suppressed the growth of Fusarium avenaceum, and 53 isolates (9%) suppressed the growth of Rhizoctonia solani CKP7. Seventeen isolates (3%) promoted canola root elongation in a growth pouch assay, and of these, 4 isolates promoted the growth of lentil and one isolate promoted the growth of pea. Fatty acid profile analysis and 16S rRNA sequencing of smaller subsets of the isolates that were positive for the plant growth-promotion traits tested showed that 39%-42% were members of the Pseudomonadaceae and 36%-42% of the Enterobacteriaceae families. Several of these isolates may have potential for development as biofertilizers or biopesticides for western Canadian legume crops.

  13. Stimulation of Carbon Dioxide Fixation in Isolated Pea Chloroplasts by Catalytic Amounts of Adenine Nucleotides 1

    PubMed Central

    Robinson, Simon P.; Wiskich, Joseph T.

    1976-01-01

    Carbon dioxide-dependent O2 evolution by isolated pea (Pisum sativum var. Massey Gem) chloroplasts was increased two to 12 times by the addition of ATP. O2 evolution was also stimulated by ADP and to a lesser extent by AMP. The ATP effects were not due to broken chloroplasts present in the preparations nor was ATP acting as a phosphate source. We concluded that the adenine nucleotides were acting catalytically. The concentration of ATP required for half-maximum rate of O2 evolution was 16 to 25 μm. The degree to which ATP stimulated O2 evolution depended on the age of pea plants from which the chloroplasts were isolated. Spinach (Spinacia oleracea var. True Hybrid 102) chloroplasts did not show a consistent stimulation of O2 evolution by adenine nucleotides. The adenine nucleotide content of pea chloroplasts was not lower than that of spinach chloroplasts, but pea chloroplasts which showed a large stimulation of O2 evolution by ATP contained an ATP-hydrolyzing reaction with rates of 10 to 50 μmol ATP hydrolyzed mg chlorophyll−1 hour−1. The rate of the ATP-consuming reaction was much lower in spinach chloroplasts and in chloroplasts from older pea plants which did not show large stimulation of O2 evolution by ATP. We propose that the ATP-consuming reaction, with a high affinity for ATP, decreased the effective size of the ATP pool available for CO2 fixation. Added adenine nucleotides could be transported into the chloroplasts increasing the concentration of internal nucleotides. Calculations showed that the adenine nucleotide transporter on the outer chloroplast membranes could operate at a sufficient rate to produce such an effect. PMID:16659638

  14. Characterization of superoxide production by isolated pea thylakoids

    SciTech Connect

    Grace, S.; Osmond, B. )

    1991-05-01

    During photosynthesis chloroplasts univalently reduce molecular oxygen to superoxide through autoxidations in the electron transport chain. Cytochrome c reduction was used to assay superoxide production in illuminate pea thylakoids under a variety of conditions. Superoxide dismutase was found to inhibit the reaction by 80%, indicating that cytochrome c reduction is primarily mediated by superoxide. This was further supported by the observation that the highest rates of cytochrome c reduction occurred in the presence of methyl viologen, an autoxidizable redox carrier that accepts electrons from photosystem I. The reaction was fully suppressed by DCMU, demonstrating a requirement for electron transport. In the presence of the plastoquinone antagonist DBMIB the rate of cytochrome c reduction increased substantially. This indicates that under conditions where electron transport to photosystem I is blocked, autoxidation reactions can occur on the reducing side of photosystem II to maintain Q{sub A} in the oxidized state. Superoxide production at sites other than the reducing side of photosystem I may thus represent an important pathway for dissipating excess excitation energy.

  15. Cadmium effects on populations of root nuclei in two pea genotypes inoculated or not with the arbuscular mycorrhizal fungus Glomus mosseae.

    PubMed

    Repetto, Ombretta; Massa, Nadia; Gianinazzi-Pearson, Vivienne; Dumas-Gaudot, Eliane; Berta, Graziella

    2007-03-01

    Plants possess a broad range of strategies to cope with cadmium (Cd) stress, including the arbuscular mycorrhizal (AM) symbiosis. In cell responses towards Cd, the contribution of changes in ploidy levels is still unclear. We used flow cytometry to investigate if nuclear ploidy changes are involved in response mechanisms toward Cd and to analyze the effect of the symbiotic status on populations of nuclei. The impact of Cd was investigated in roots of two pea (Pisum sativum L.) genotypes differing in their Cd-sensitivity (Cd-sensitive VIR4788 and Cd-tolerant VIR7128). In pea seedlings grown under hydropony, 25 and 250 microM Cd concentrations lead to an increase in 4 C together with a decrease in 2 C nuclei. The same genotypes, grown in soil/sand substrate, were inoculated or not with the AM fungus Glomus mosseae BEG12 and treated or not with Cd at transplanting (Cd1) or 2 weeks after (Cd2). The Cd2 increased the proportion of 6 and 8 C nuclei in the mycorrhizal VIR4788 and in the non-mycorrhizal VIR7128 genotypes. Thus, changes in ploidy levels reflect pea responses towards Cd, which are modulated by the symbiotic interaction. The Cd-induced increase in ploidy may account for changes in DNA transcription and/or translation.

  16. Partial purification and characterization of an enzyme from pea nuclei with protein tyrosine phosphatase activity.

    PubMed Central

    Guo, Y L; Roux, S J

    1995-01-01

    A pea (Pisum sativum L.) nuclear enzyme with protein tyrosine phosphatase activity has been partially purified and characterized. The enzyme has a molecular mass of 90 kD as judged by molecular sieve column chromatography and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Like animal protein tyrosine phosphatases it can be inhibited by low concentrations of molybdate and vanadate. It is also inhibited by heparin and spermine but not by either the acid phosphatase inhibitors citrate and tartrate or the protein serine/threonine phosphatase inhibitor okadaic acid. The enzyme does not require Ca2+, Mg2+, or Mn2+ for its activity but is stimulated by ethylenediaminetetraacetate and by ethyleneglycolbis(beta-aminoethyl ether)-N,N'-tetraacetic acid. It dephosphorylates phosphotyrosine residues on the four different 32P-tyrosine-labeled peptides tested but not the phosphoserine/threonine residues on casein and histone. Like some animal protein tyrosine phosphatases, it has a variable pH optimum depending on the substrate used: the optimum is 5.5 when the substrate is [32P]tyrosine-labeled lysozyme, but it is 7.0 when the substrate is [32P]tyrosine-labeled poly(glutamic acid, tyrosine). It has a Km of 4 microM when the lysozyme protein is used as a substrate. PMID:11536662

  17. Emulsification properties of pea protein isolate using homogenization, microfluidization and ultrasonication.

    PubMed

    McCarthy, Noel A; Kennedy, Deirdre; Hogan, Sean A; Kelly, Philip M; Thapa, Krishtina; Murphy, Kevin M; Fenelon, Mark A

    2016-11-01

    Pea protein isolate (PPI) is used in many food formulations, due to its low cost, commercial availability and excellent amino acid profile. The objective of this study was to determine the emulsification properties of PPI. Particle size of PPI powders showed neither temperature (25-65°C) nor time (up to 24h) increased solubilisation of powder particles during mixing. Heating PPI dispersions (10%, w/w, protein) from 45 to 90°C led to an increase in storage modulus (G'; Pa) at 71°C, indicating the onset of protein aggregation. Gel formation occurred at 79°C (G'>1Pa). Pea protein-stabilised emulsions made using homogenization (15MPa; 1 pass) or microfluidization (50MPa; 1 pass) resulted in the formation of cold-set gels, with gel strength increasing with increasing oil concentration and fluidic pressure. Droplet size and viscosity of pea protein-stabilised emulsions decreased and increased, respectively, with increasing ultrasonication time. Overall, ultrasonication (<50°C) can create a uniform droplet size emulsion, while, homogenization and microfluidization can produce cold-set gels for use in a wide-range of food applications. Copyright © 2016. Published by Elsevier Ltd.

  18. Protocol to cryopreserve and isolate nuclei from adipose tissue without dimethyl sulfoxide.

    PubMed

    Almeida, M M; Caires, L C J; Musso, C M; Campos, J M S; Maranduba, C M C; Macedo, G C; Mendonça, J P R F; Garcia, R M G

    2014-12-19

    Cryopreservation injuries involve nuclear DNA damage. A protocol for cryopreserving and isolating adipocyte nuclei is proposed. Adipose tissue samples were directly analyzed (NoCRYO-0h), or stored at -196°C for 7 days without 10% dimethyl sulfoxide (DMSO) (CRYO-WO-DMSO) or with DMSO (CRYO-W-DMSO). To determine the effect of DMSO on cryopreservation treatment, adipose tissue samples were stored at 4°C for 24 h with 10% DMSO (NoCRYO-W-DMSO-24h) and without (NoCRYO-WO-DMSO-24h). Samples were processed in isolation buffer, and nuclear integrity was measured by flow cytometry. The coefficient of variation, forward scatter, side scatter, and number of nuclei analyzed were evaluated. Pea (Pisum sativum) was used to measure the amount of DNA. All groups contained similar amounts of DNA to previously reported values and a satisfactory number of nuclei were analyzed. CRYO-W-DMSO presented a higher coefficient of variation (3.19 ± 0.09) compared to NoCRYO-0h (1.85 ± 0.09) and CRYO-WO-DMSO (2.02 ± 0.02). The coefficient of variation was increased in NoCRYO-W-DMSO-24h (3.80 ± 0.01) compared to NoCRYO-WO-DMSO-24h (2.46 ± 0.03). These results relate DMSO presence to DNA damage independently of the cryopreservation process. CRYO-W-DMSO showed increased side scatter (93.46 ± 5.03) compared to NoCRYO-0h (41.13 ± 3.19) and CRYO-WO-DMSO (48.01 ± 2.28), indicating that cryopreservation with DMSO caused chromatin condensation and/or nuclear fragmentation. CRYO-W-DMSO and CRYO-WO-DMSO presented lower forward scatter (186.33 ± 9.33 and 196.89 ± 26.86, respectively) compared to NoCRYO-0h (322.80 ± 3.36), indicating that cryopreservation reduced nuclei size. Thus, a simple method for cryopreservation and isolation of adipocyte nuclei causing less damage to DNA integrity was proposed.

  19. Light regulation of the abundance of mRNA encoding a nucleolin-like protein localized in the nucleoli of pea nuclei.

    PubMed Central

    Tong, C G; Reichler, S; Blumenthal, S; Balk, J; Hsieh, H L; Roux, S J

    1997-01-01

    A cDNA encoding a nucleolar protein was selected from a pea (Pisum sativum) plumule library, cloned, and sequenced. The translated sequence of the cDNA has significant percent identity to Xenopus laevis nucleolin (31%), the alfalfa (Medicago sativa) nucleolin homolog (66%), and the yeast (Saccharomyces cerevisiae) nucleolin homolog (NSR1) (28%). It also has sequence patterns in its primary structure that are characteristic of all nucleolins, including an N-terminal acidic motif, RNA recognition motifs, and a C-terminal Gly- and Arg-rich domain. By immunoblot analysis, the polyclonal antibodies used to select the cDNA bind selectively to a 90-kD protein in purified pea nuclei and nucleoli and to an 88-kD protein in extracts of Escherichia coli expressing the cDNA. In immunolocalization assays of pea plumule cells, the antibodies stained primarily a region surrounding the fibrillar center of nucleoli, where animal nucleolins are typically found. Southern analysis indicated that the pea nucleolin-like protein is encoded by a single gene, and northern analysis showed that the labeled cDNA binds to a single band of RNA, approximately the same size and the cDNA. After irradiation of etiolated pea seedlings by red light, the mRNA level in plumules decreased during the 1st hour and then increased to a peak of six times the 0-h level at 12 h. Far-red light reversed this effect of red light, and the mRNA accumulation from red/far-red light irradiation was equal to that found in the dark control. This indicates that phytochrome may regulate the expression of this gene. PMID:9193096

  20. Replication of DNA by Nuclei Isolated from Soybean Suspension Cultures 1

    PubMed Central

    Roman, Ruth; Caboche, Michel; Lark, Karl G.

    1980-01-01

    DNA replication was studied in nuclei isolated from soybean cells grown in suspension culture. The isolation procedure involved the preparation of protoplasts, their lysis with a nonionic detergent and purification of nuclei. These nuclei synthesized low molecular weight DNA and joined these fragments into DNA of intermediate molecular weight. The characteristics of replication in isolated nuclei correlated well with those of the cells from which they were isolated, as shown by fluorodeoxyuridine synchronization and ultraviolet irradiation experiments. Images PMID:16661510

  1. Isolation and Characterization of an Alkaline Phosphatase from Pea Thylakoids 1

    PubMed Central

    Kieleczawa, Jan; Coughlan, Sean J.; Hind, Geoffrey

    1992-01-01

    Endogenous dephosphorylation of the light-harvesting chlorophyll-protein complex of photosystem II in pea (Pisum sativum, L. cv Progress 9) thylakoids drives the state 2 to state 1 transition; the responsible enzyme is a thylakoid-bound, fluoride-sensitive phosphatase with a pH optimum of 8.0 (Bennett J [1980] Eur J Biochem 104: 85-89). An enzyme with these characteristics was isolated from well-washed thylakoids. Its molecular mass was estimated at 51.5 kD, and this monomer was catalytically active, although the activity was labile. The active site could be labeled with orthophosphate at pH 5.0. High levels of alkaline phosphatase activity were obtained with the assay substrate, 4-methylumbelliferyl phosphate (350 micromoles per minute per milligram purified enzyme). The isolated enzyme functioned as a phosphoprotein phosphatase toward phosphorylated histone III-S and phosphorylated, photosystem II-enriched particles from pea, with typical activities in the range of 200 to 600 picomoles per minute per milligram enzyme. These activities all had a pH optimum of 8.0 and were fluoride sensitive. The enzyme required magnesium ion for maximal activity but was not dependent on this ion. Evidence supporting a putative function for this phosphatase in dephosphorylation of thylakoid proteins came from the inhibition of this process by a polyclonal antibody preparation raised against the partially purified enzyme. ImagesFigure 2Figure 3 PMID:16668967

  2. Rapid Isolation of Nuclei from Cells In Vitro.

    PubMed

    Nabbi, Arash; Riabowol, Karl

    2015-08-03

    This protocol presents a rapid, efficient, and practical (REAP) method to separate nuclei from cultured cells in vitro with as little damage and contamination as possible. The REAP procedure is performed at low temperature and takes <2 min, which minimizes protein degradation, protein modification, and diffusion of soluble proteins out of the nuclear compartment while maintaining the integrity of protein complexes. A mild detergent, NP-40, is used together with mild mechanical shearing to disrupt the plasma membrane, leaving the nuclear membrane intact. The REAP method can be used with various cell lines grown in vitro and requires minimal optimization. The isolated nuclei are suitable for numerous downstream applications (e.g., western blotting, 2D gel electrophoresis, and immunoprecipitation). If desired, aliquots of whole-cell lysate and the cytoplasmic fraction can be saved for comparison.

  3. Molecular characterization and identification of plant growth promoting endophytic bacteria isolated from the root nodules of pea (Pisum sativum L.).

    PubMed

    Tariq, Mohsin; Hameed, Sohail; Yasmeen, Tahira; Zahid, Mehwish; Zafar, Marriam

    2014-02-01

    Root nodule accommodates various non-nodulating bacteria at varying densities. Present study was planned to identify and characterize the non-nodulating bacteria from the pea plant. Ten fast growing bacteria were isolated from the root nodules of cultivated pea plants. These bacterial isolates were unable to nodulate pea plants in nodulation assay, which indicate the non-rhizobial nature of these bacteria. Bacterial isolates were tested in vitro for plant growth promoting properties including indole acetic acid (IAA) production, nitrogen fixation, phosphate solubilization, root colonization and biofilm formation. Six isolates were able to produce IAA at varying level from 0.86 to 16.16 μg ml(-1), with the isolate MSP9 being most efficient. Only two isolates, MSP2 and MSP10, were able to fix nitrogen. All isolates were able to solubilize inorganic phosphorus ranging from 5.57 to 11.73 μg ml(-1), except MSP4. Bacterial isolates showed considerably better potential for colonization on pea roots. Isolates MSP9 and MSP10 were most efficient in biofilm formation on polyvinyl chloride, which indicated their potential to withstand various biotic and abiotic stresses, whereas the remaining isolates showed a very poor biofilm formation ability. The most efficient plant growth promoting agents, MSP9 and MSP10, were phylogenetically identified by 16S rRNA gene sequence analysis as Ochrobactrum and Enterobacter, respectively, with 99% similarity. It is suggested the potential endophytic bacterial strains, Ochrobactrum sp. MSP9 and Enterobacter sp. MSP10, can be used as biofertilizers for various legume and non-legume crops after studying their interaction with the host crop and field evaluation.

  4. The effect of germination on antioxidant and nutritional parameters of protein isolates from grass pea (Lathyrus sativus) seeds.

    PubMed

    Starzyńska-Janiszewska, A; Stodolak, B; Mickowska, B

    2010-02-01

    The aim of this research was to study the antioxidant and nutritional (selected objects) properties of protein isolates obtained from grass pea seedlings as compared with soaked and raw seeds. Two percent extract of isolate from 5-day-old seedlings showed the highest total antioxidant activity (25%) and the ability to chelate Fe²+ (2.35 mg/g d.m.) as compared with other isolates. Protein isolates from grass pea seeds had on average 89% total protein, 87% in vitro protein bioavailability, about 5574 TIU/g (d.m.) (trypsin inhibitors activity) and did not contain ODAP. Germination of seeds for 5 days considerably improved the in vitro bioavailability of isolates, by 12%, and profile of sulfur amino acids by 42%, in comparison with isolates obtained from the raw seeds. Isolates from 5-day-old grass pea seedlings had the best antioxidant properties and improved nutritional parameters (as compared with raw seeds), which makes them worthy of being considered as a potential food additive.

  5. Uptake of auxins into membrane vesicles isolated from pea stems: an in vitro auxin transport system

    SciTech Connect

    Slone, J.H.

    1985-01-01

    The objective of this research was to test the applicability of the chemiosmotic theory of auxin transport to a subcellular system. Membrane vesicles were isolated from the basal portion of the third internode of etiolated pea plants (Pisum sativum L. var. Alaska) by differential centrifugation. Uptake of auxin was determined by adding /sup 14/C-labeled indoleacetic acid (IAA) to vesicles. Nigericin, a monovalent cation ionophore, and the electrogenic protonophore, carbonyl-cyanide m-chlorophenylhydrazone (CCCP), at micromolar concentrations abolished saturable uptake. Bursting vesicles by sonication, osmotic shock and freeze/thawing also eliminated saturable uptake. As the temperature increased from 0 to 30/sup 0/C, saturable uptake decreased markedly. Nonsaturable auxin uptake was less affected by these treatments. The pH gradient-dependent uptake of auxin appeared to be a transmembrane uptake of auxin into the vesicles rather than surface binding. Unlabeled IAA, 2,4-dichlorophenoxyacetic acid (2,4-D) and 2-naphthaleneacetic acid (NAA) at low concentrations reduced the saturable accumulation of (/sup 14/C)IAA in vesicles, while phenylacetic acid, benzoic acid, and 1-NAA were effective only at high concentrations. Kinetic analysis revealed two types of sites: a high affinity site with an uptake capacity of 25 to 40 pmoles/g tissue, and a low affinity site with an uptake capacity of 260 to 600 pmole/g tissue, fresh wt. In conclusion, several principal elements of an auxin transport system, as specific by the chemiosmotic theory of polar auxin transport, were present in membrane vesicles isolated from relatively mature pea stem tissue. However, one important aspect of the theory was not demonstrated in this in vitro system - a TIBA/NPA-sensitive auxin efflux. The kinetics and specificity of auxin uptake strongly suggested that this system was physiologically significant.

  6. Isolation of pigment-binding early light-inducible proteins from pea.

    PubMed

    Adamska, I; Roobol-Bóza, M; Lindahl, M; Andersson, B

    1999-03-01

    The early light-inducible proteins (ELIPs) in chloroplasts possess a high sequence homology with the chlorophyll a/b-binding proteins but differ from those proteins by their substoichiometric and transient appearance. In the present study ELIPs of pea were isolated by a two-step purification strategy: perfusion chromatography in combination with preparative isoelectric focussing. Two heterogeneous populations of ELIPs were obtained after chromatographic separation of solubilized thylakoid membranes using a weak anion exchange column. One of these populations contained ELIPs in a free form providing the first isolation of these proteins. To prove whether the isolated and pure forms of ELIP bind pigments, spectroscopic and chromatographic analysis were performed. Absorption spectra and TLC revealed the presence of chlorophyll a and lutein. Measurements of steady-state fluorescence emission spectra at 77 K exhibited a major peak at 674 nm typical for chlorophyll a bound to the protein matrix. The action spectrum of the fluorescence emission measured at 674 nm showed several peaks originating mainly from chlorophyll a. It is proposed that ELIPs are transient chlorophyll-binding proteins not involved in light-harvesting but functioning as scavengers for chlorophyll molecules during turnover of pigment-binding proteins.

  7. Light-dependent degradation of the QB-protein in isolated pea thylakoids

    PubMed Central

    Ohad, I.; Kyle, D. J.; Hirschberg, J.

    1985-01-01

    The 32 000-dalton QB-protein of photosystem II (PS II) is rapidly damaged and removed from isolated pea thylakoids during incubation in the light resulting in a loss of photosynthetic electron flow through PS II. This in vitro photoinhibition is similar to that previously reported with intact Chlamydomonas cells. The damage occurs at a faster rate in vitro, however, due to the inability of isolated thylakoids to synthesize replacement QB-protein. The removal of the damaged QB-protein does not require any soluble components of the chloroplast stroma and is unaffected by the protease inhibitors phenyl-methylsulfonylfluoride or antipain. Unlike the effect of trypsin, no low mol. wt. membrane-bound or soluble fragments of the labelled QB-protein could be identified either by autoradiography or immunologically using polyclonal antibodies specific for the QB-protein. The lightinduced damage to the QB-protein (indicated by a loss of QB functional activity), preceded the removal of the protein from the membrane. We conclude that photodamage of the QB-protein generates a conformational change which renders the protein susceptible to attack by a highly efficient, intrinsic membrane protease. ImagesFig. 1.Fig. 3.Fig. 4.Fig. 5. PMID:16453621

  8. The isolation of nuclei containing mitotic spindles from the plasmodium of the slime mould Physarum polycephalum.

    PubMed

    Roobol, A; Paul, E C; Gull, K

    1984-05-01

    We have developed a protocol which facilitates the isolation of nuclei containing mitotic spindles from the plasmodium of Physarum polycephalum. The nuclei are isolated in a spindle stabilization buffer (SSB) consisting of -4 M glycerol, 5 mM EGTA, 10 mM MgCl2, 10 mM PIPES and 0.1% (v/v) Triton X-100 at pH 6.5. Isolation at 20 degrees C, chelation of Ca2+ and the inclusion of the 4 M glycerol were found to be essential for stabilization of the spindle microtubules. The method allows the isolation of clean nuclei in a reasonable yield. Nuclei were isolated at each of the main stages of mitosis (prophase, metaphase and anaphase) and transmission electron microscopy of the isolated nuclei revealed well preserved arrangements of spindle microtubules which were characteristic of the particular stage of mitosis.

  9. Effects of GA sub 4 on gene expression in isolated nuclei

    SciTech Connect

    Sechley, K.A.; Srivastava, L.M. )

    1989-04-01

    GA{sub 4} is active in promoting elongation in cucumber (Cucuminus sativus) hypocotyls. Nuclei, isolated from the apical 1cm portion of the hypocotyl and purified on Percoll step gradients increased transcription (({sup 3}H)UTP incorporation into TCA precipitable products) in the presence of GA{sub 4} compared to untreated nuclei. This response was not observed in nuclei isolated from basal (nontarget) regions of the hypocotyl. Enhanced transcription in the presence of GA{sub 4} appears to be affected by proteinaceous factors which can be washed out of the nuclei. The GA{sub 4} induced response of cucumber hypocotyls is temperature sensitive; nuclei isolated from plants grown at 34C do not show the above GA{sub 4}-induced transcriptional enhancement. Comparison of 2-D flurograms of in vitro translation products synthesized from transcripts obtained from intact plants and isolated nuclei in the presence or absence of GA{sub 4} is currently in progress.

  10. Functional and pasting properties of pea starch and peanut protein isolate blends.

    PubMed

    Sun, Qingjie; Xiong, Cuixia Sun Liu

    2014-01-30

    The functional and pasting properties of pea starch (PS) and peanut protein isolate (PPI) blends mixed at different proportions were studied. With the increasing ratio of PPI in PS/PPI blends, the solubility of the blends determined at 90°C was increased from 16.38 to 31.28% whereas both of the water absorption capacity and the swelling power decreased. The pasting temperature of the blends increased from 72.5 to 77.5°C while the peak viscosity decreased from 276.33 to 39.92 RVU upon the increasing level of PPI. The hardness of the PS/PPI blends gel decreased from 9.67 N to 0.96 N when the PPI content was increased from 0 to 50% in the blend. Scanning electron microscopy exhibited a honeycomb feature at the ratio of 90:10 and 80:20. The large fragmentary structure of the blending gels was formed at the ratio of 70:30 and became more loosed with the increased ratio of PPI in the blends.

  11. Synthesis of a Putative c-Type Cytochrome by Intact, Isolated Pea Chloroplast.

    PubMed

    Bhaya, D; Castelfranco, P A

    1986-08-01

    In addition to chlorophyll-protein complexes, other proteins were labeled when isolated developing pea (Pisum sativum L.) chloroplasts were incubated with [(14)C]-5-aminolevulinic acid. The major labeled band (M(r) = 43 kilodaltons by lithium dodecyl sulfate-polyacrylamide gel electrophoresis) was labeled even in the presence of chloramphenicol. Heme-dependent peroxidase activity (as detected by the tetramethyl benzidine-H(2)O(2) stain) was not visibly associated with this band. The radioactive band was stable to heat, 5% HCl in acetone, and was absent if the incubation with [(14)C]-5-aminolevulinic acid was carried out in the presence of N-methyl protoporphyrin IX dimethyl ester (a specific inhibitor of ferrochelatase). Organic solvent extraction procedures for the enrichment of cytochrome f from chloroplast membranes also extracted this unknown labeled product. It was concluded that this labeled product was probably a c-type cytochrome; however, the possibility that it might be a protein containing a covalently linked linear tetrapyrrole was not ruled out.

  12. The pH dependence of violaxanthin deepoxidation in isolated pea chloroplasts

    SciTech Connect

    Pfuendel, E.E.; Dilley, R.A. )

    1993-01-01

    The absorbance change at 505 nm was used to monitor the kinetics of violaxanthin deepoxidation in isolated pea (Pisum sativum) chloroplasts under dark conditions at various pH values. In long-term measurements (65 min) a fast and a slow exponential component of the 505-nm absorbance change could be resolved. The fast rate constant was up to 10 times higher than the slow rate constant. The asymptote value of the fast kinetic component was twice that of the slow component. The pH dependency of the parameters of the fast kinetic component was analyzed from pH 5.2 to pH 7.0. It was found that the asymptote value dropped slightly with increasing pH. The rate constant was zero at pH values greater than 6.3 and showed maximum values at pH values less than 5.8. Hill plot analysis revealed a strong positive cooperativity for the pH dependency of the fast rate constant (Hill coefficient n[sub H] = 5.3). The results are discussed with respect to published activity curves of violaxanthin deepoxidation. 23 refs., 6 figs., 1 tab.

  13. FACTORS INFLUENCING THE ABILITY OF ISOLATED CELL NUCLEI TO FORM GELS IN DILUTE ALKALI

    PubMed Central

    Dounce, Alexander L.; Monty, Kenneth J.

    1955-01-01

    1. Known methods for isolating cell nuclei are divided into two classes, depending on whether or not the nuclei are capable of forming gels in dilute alkali or strong saline solutions. Methods which produce nuclei that can form gels apparently prevent the action of an intramitochondrial enzyme capable of destroying the gel-forming capacity of the nuclei. Methods in the other class are believed to permit this enzyme to act on the nuclei during the isolation procedure, causing detachment of DNA from some nuclear constituent (probably protein). 2. It is shown that heating in alkaline solution and x-irradiation can destroy nuclear gels. Heating in acid or neutral solutions can destroy the capacity of isolated nuclei to form gels. 3. Chemical and biological evidence is summarized in favor of the hypothesis that DNA is normally bound firmly to some nuclear component by non-ionic linkages. PMID:14381437

  14. Evidence for Mediated HCO3− Transport in Isolated Pea Mesophyll Protoplasts 1

    PubMed Central

    Volokita, Micha; Kaplan, Aaron; Reinhold, Leonora

    1981-01-01

    The kinetics of 14C fixation, and inorganic C (Cinorg) accumulation, have been followed in isolated pea mesophyll protoplasts. NaH14CO3 was supplied to the protoplasts in media the pH of which was varied between 7 and 8. When 14CO2 fixation was plotted against the calculated concentration of free CO2 in the media, the apparent Km for CO2 was observed to rise as external pH increased. The Vmax did not alter significantly. Similarly, when Cinorg uptake, either in the light or in the dark, was plotted against external CO2 concentration the slope of the curves was steeper at higher external pH. Investigation of the time course of uptake showed that internal Cinorg concentration rose throughout the experimental period, and that in the light it surpassed the external Cinorg concentration after about 3 minutes. Irradiation of protoplasts previously taking up 14Cinorg in the dark brought about a sharp increase in the rate of 14Cinorg accumulation which was sustained for at least 20 minutes. Estimates of internal pH based on the distribution of labeled 5,5-dimethyloxazoladine-2,4-dione (DMO) between protoplast and medium suggested that internal pH altered relatively little with change in external pH. The values for internal pH as calculated from Cinorg distribution were always higher than those calculated from DMO distribution, i.e. the internal Cinorg concentration was higher than would be predicted on the assumption of passive distribution in accordance with pH. Addition of carbonic anhydrase to the external solution was without effect either on rate of 14CO2 fixation or Cinorg accumulation. Various possible interpretations of the results are considered. It is concluded that the most reasonable explanation, consistent with all the data, is that HCO3− ions can cross the protoplast membranes, and that their passage is mediated by a transfer mechanism. PMID:16661821

  15. Heteroprotein Complex Formation of Bovine Lactoferrin and Pea Protein Isolate: A Multiscale Structural Analysis.

    PubMed

    Adal, Eda; Sadeghpour, Amin; Connell, Simon; Rappolt, Michael; Ibanoglu, Esra; Sarkar, Anwesha

    2017-02-13

    Associative electrostatic interactions between two oppositely charged globular proteins, lactoferrin (LF) and pea protein isolate (PPI), the latter being a mixture of vicilin, legumin, and convicilin, was studied with a specific PPI/LF molar ratio at room temperature. Structural aspects of the electrostatic complexes probed at different length scales were investigated as a function of pH by means of different complementary techniques, namely, with dynamic light scattering, small-angle X-ray scattering (SAXS), turbidity measurements, and atomic force microscopy (AFM). Irrespective of the applied techniques, the results consistently displayed that complexation between LF and PPI did occur. In an optimum narrow range of pH 5.0-5.8, a viscous liquid phase of complex coacervate was obtained upon mild centrifugation of the turbid LF-PPI mixture with a maximum Rh, turbidity and the ζ-potential being close to zero observed at pH 5.4. In particular, the SAXS data demonstrated that the coacervates were densely assembled with a roughly spherical size distribution exhibiting a maximum extension of ∼80 nm at pH 5.4. Equally, AFM image analysis showed size distributions containing most frequent cluster sizes around 40-80 nm with spherical to elliptical shapes (axis aspect ratio ≤ 2) as well as less frequent elongated to chainlike structures. The most frequently observed compact complexes, we identify as mainly leading to LF-PPI coacervation, whereas for the less frequent chain-like aggregates, we hypothesize that additionally PPI-PPI facilitated complexes exist.

  16. Intrathylakoid pH in Isolated Pea Chloroplasts as Probed by Violaxanthin Deepoxidation.

    PubMed Central

    Pfundel, E. E.; Renganathan, M.; Gilmore, A. M.; Yamamoto, H. Y.; Dilley, R. A.

    1994-01-01

    Light-driven violaxanthin deepoxidation was measured in isolated pea (Pisum sativum) chloroplasts without ATP synthesis (basal conditions) and with ATP synthesis (coupled conditions). Thylakoids stored in high salt (HS) or low salt (LS) storage medium were tested. In previous experiments, HS thylakoids and LS thylakoids were related to delocalized and localized proton coupling, respectively.Light-driven deepoxidase activity was compared to the pH dependence of deepoxidase activity established in dark reactions. At an external pH of 8, light-driven deepoxidation indicated effective pH values close to pH 6 for all reaction conditions. Parallel to deepoxidation, the thylakoid lumen pH was estimated by the fluorescent dye pyranine.In LS thylakoids under coupled conditions the lumen pH did not drop below pH 6.7. At pH 6.7, no deepoxidase activity is expected based on the pH dependence of enzyme activity. The results suggest that deepoxidation activity is controlled by the pH in sequestered membrane domains, which, under localized proton coupling, can be maintained at pH 6.0 when the lumen pH is far above pH 6.0. The extent of violaxanthin conversion (availability), however, appeared to be regulated by lumenal pH. Dithiothreitol-sensitive nonphotochemical quenching of chlorophyll fluorescence was dependent on zeaxanthin and not related to lumenal pH. Thus, zeaxanthin-dependent quenching[mdash]known to be pH dependent[mdash]appeared to be triggered by the pH of localized membrane domains. PMID:12232439

  17. Characteristics of DNA replication in isolated nuclei initiated by an aprotinin-binding protein.

    PubMed

    Coffman, F D; Fresa, K L; Hameed, M; Cohen, S

    1993-02-01

    Isolated cell nuclei were used as the source of template DNA to investigate the role of a cytosolic aprotinin-binding protein (ADR) in the initiation of eukaryotic DNA replication. Computerized image cytometry demonstrated that the DNA content of individual nuclei increased significantly following incubation with ADR-containing preparations, and the extent of DNA synthesis is consistent with that allowed by the limiting concentration of dTTP. Thus, dTTP incorporation into isolated nuclei represents DNA synthesis and not parent strand repair. We found that dTTP incorporation into the isolated nuclei is dependent on DNA polymerase alpha (a principal polymerase in DNA replication) but that DNA polymerase beta (a principal polymerase in DNA repair processes) does not play a significant role in this system. Finally, neither aprotinin nor a previously described cytosolic ADR inhibitor can block the replication of nuclease-treated calf thymus DNA, while both strongly inhibit replication of DNA in isolated nuclei. This result, coupled with the relative ineffectiveness of nuclease-treated DNA compared with nuclear DNA to serve as a replicative template in this assay, argues against a significant contribution from repair or synthesis which initiates at a site of DNA damage. These data indicate that ADR-mediated incorporation of 3H-dTTP into isolated nuclei results from DNA replicative processes that are directly relevant to in vivo S phase events.

  18. Nucleotide sequence and infectious cDNA clone of the L1 isolate of Pea seed-borne mosaic potyvirus.

    PubMed

    Olsen, B S; Johansen, I E

    2001-01-01

    The complete nucleotide sequence of Pea seed-borne mosaic potyvirus isolate L1 has been determined from cloned virus cDNA. The PSbMV L1 genome is 9895 nucleotides in length excluding the poly(A) tail. Computer analysis of the sequence revealed a single long open reading frame (ORF) of 9594 nucleotides. The ORF potentially encodes a polyprotein of 3198 amino acids with a deduced Mr of 363537. Nine putative proteolytic cleavage sites were identified by analogy to consensus sequences and genome arrangement in other potyviruses. Two full-length cDNA clones, p35S-L1-4 and p35S-L1-5, were assembled under control of an enhanced 35S promoter and nopaline synthase terminator. Clone p35S-L1-4 was constructed with four introns and p35S-L1-5 with five introns inserted in the cDNA. Clone p35S-L1-4 was unstable in Escherichia coli often resulting in amplification of plasmids with deletions. Clone p35S-L1-5 was stable and apparently less toxic to Escherichia coli resulting in larger bacterial colonies and higher plasmid yield. Both clones were infectious upon mechanical inoculation of plasmid DNA on susceptible pea cultivars Fjord, Scout, and Brutus. Eight pea genotypes resistant to L1 virus were also resistant to the cDNA derived L1 virus. Both native PSbMV L1 and the cDNA derived virus infected Chenopodium quinoa systemically giving rise to characteristic necrotic lesions on uninoculated leaves.

  19. Improving functional properties of pea protein isolate for microencapsulation of flaxseed oil.

    PubMed

    Bajaj, Poonam R; Bhunia, Kanishka; Kleiner, Leslie; Joyner Melito, Helen S; Smith, Denise; Ganjyal, Girish; Sablani, Shyam S

    2017-04-10

    Unhydrolyzed pea protein (UN) forms very viscous emulsions when used at higher concentrations. To overcome this, UN was hydrolyzed using enzymes Alcalase, Flavourzyme, Neutrase, Alcalase-Flavourzyme and Neutrase-Flavourzyme at 50°C for 0 min, 30 min, 60 min and 120 min to form hydrolyzed proteins A, F, N, AF and NF, respectively. All hydrolyzed proteins had lower apparent viscosity and higher solubility than UN. Foaming capacity of A was highest, followed by NF, N, and AF. Hydrolyzed proteins N60, A60, NF60 and AF60 were prepared by hydrolyzing UN for 60 min and used further for microencapsulation. At 20% oil loading (on a total solid basis), the encapsulated powder N60 had the highest microencapsulation efficiency (ME = 56.2). A decrease in ME occurred as oil loading increased to 40%. To improve the ME of N60, >90%, UN and maltodextrin were added. Flowability and particle size distribution of microencapsulated powders with >90% microencapsulation efficiency and morphology of all powders were investigated. This study identified a new way to improve pea protein functionality in emulsions, as well as a new application of hydrolyzed pea protein as wall material for microencapsulation.

  20. Biochemical and immunological characterization of pea nuclear intermediate filament proteins.

    PubMed

    Blumenthal, Sonal S D; Clark, Gregory B; Roux, Stanley J

    2004-04-01

    In immunoblot assays, at least three putative nuclear intermediate filament (NIF) proteins were detected in nuclear envelope-matrix (NEM) and lamin (L1) fractions of nuclei from plumules of dark-grown pea (Pisum sativum L.) seedlings. These NIF proteins had apparent molecular masses of ca. 65, 60, and 54 kDa (also referred to as p65, p60, and p54), and appeared as multiple isoelectric forms, with pIs ranging from ca. 4.8 to 6.0. Polyclonal and monoclonal antibodies were raised to the 65-kDa NIF protein bands excised from gels after electrophoresis. These anti-pea antibodies were specifically cross-reactive with the pea nuclear p65, p60, and p54 proteins and also with chicken lamins. Sequence alignment of peptide fragments obtained from the 65- and 60-kDa pea NIF proteins showed similarity with animal intermediate filament proteins such as lamins and keratins and with certain plant proteins predicted to have long coiled-coil domains. These pea NIF proteins were further purified and enriched from the NEM fraction using methods similar to those used for isolating animal lamins. When negatively stained and viewed by transmission electron microscopy, the filaments in the pea lamin (L1) fraction appeared to be 6-12 nm in diameter. As assayed by immunofluorescence cytochemistry using a confocal laser-scanning microscope, fixed pea plumule cells displayed uniform as opposed to peripheral nuclear staining by several of the antibody preparations, both polyclonal and monoclonal. This report describes the biochemical and immunological properties of these pea NIF proteins.

  1. Novel nuclei isolation buffer for flow cytometric genome size estimation of Zingiberaceae: a comparison with common isolation buffers.

    PubMed

    Sadhu, Abhishek; Bhadra, Sreetama; Bandyopadhyay, Maumita

    2016-11-01

    Cytological parameters such as chromosome numbers and genome sizes of plants are used routinely for studying evolutionary aspects of polyploid plants. Members of Zingiberaceae show a wide range of inter- and intrageneric variation in their reproductive habits and ploidy levels. Conventional cytological study in this group of plants is severely hampered by the presence of diverse secondary metabolites, which also affect their genome size estimation using flow cytometry. None of the several nuclei isolation buffers used in flow cytometry could be used very successfully for members of Zingiberaceae to isolate good quality nuclei from both shoot and root tissues. The competency of eight nuclei isolation buffers was compared with a newly formulated buffer, MB01, in six different genera of Zingiberaceae based on the fluorescence intensity of propidium iodide-stained nuclei using flow cytometric parameters, namely coefficient of variation of the G0/G1 peak, debris factor and nuclei yield factor. Isolated nuclei were studied using fluorescence microscopy and bio-scanning electron microscopy to analyse stain-nuclei interaction and nuclei topology, respectively. Genome contents of 21 species belonging to these six genera were determined using MB01. Flow cytometric parameters showed significant differences among the analysed buffers. MB01 exhibited the best combination of analysed parameters; photomicrographs obtained from fluorescence and electron microscopy supported the superiority of MB01 buffer over other buffers. Among the 21 species studied, nuclear DNA contents of 14 species are reported for the first time. Results of the present study substantiate the enhanced efficacy of MB01, compared to other buffers tested, in the generation of acceptable cytograms from all species of Zingiberaceae studied. Our study facilitates new ways of sample preparation for further flow cytometric analysis of genome size of other members belonging to this highly complex polyploid family.

  2. Isolated plant nuclei as mechanical and thermal sensors involved in calcium signalling.

    PubMed

    Xiong, Tou Cheu; Jauneau, Alain; Ranjeva, Raoul; Mazars, Christian

    2004-10-01

    Calcium signals in the nucleus elicit downstream effects that are distinct from those of cytosolic calcium signals. In the present work, we have evaluated the ability of plant nuclei to sense stimuli directly and to convert them into calcium changes. We show that individual mechanical stimulation of isolated nuclei elicits a single calcium transient at acidic pHs, whereas a series of stimulations leads to oscillations whose frequency reflects that of the stimuli. Conversely, at alkaline pHs, nuclei respond to temperature but not to stretch. The stretch- and the temperature-activated processes differ by their sensitivity to pharmacological drugs known to affect ion channel activities in animal cells. Our data demonstrate that isolated nuclei are able to gauge physical parameters of their environment. This might have a profound influence on the functioning of calcium-dependent processes known to control a large array of molecular events in the nucleus.

  3. Volume Transitions of Isolated Cell Nuclei Induced by Rapid Temperature Increase.

    PubMed

    Chan, Chii J; Li, Wenhong; Cojoc, Gheorghe; Guck, Jochen

    2017-03-28

    Understanding the physical mechanisms governing nuclear mechanics is important as it can impact gene expression and development. However, how cell nuclei respond to external cues such as heat is not well understood. Here, we studied the material properties of isolated nuclei in suspension using an optical stretcher. We demonstrate that isolated nuclei regulate their volume in a highly temperature-sensitive manner. At constant temperature, isolated nuclei behaved like passive, elastic and incompressible objects, whose volume depended on the pH and ionic conditions. When the temperature was increased suddenly by even a few degrees Kelvin, nuclei displayed a repeatable and reversible temperature-induced volume transition, whose sign depended on the valency of the solvent. Such phenomenon is not observed for nuclei subjected to slow heating. The transition temperature could be shifted by adiabatic changes of the ambient temperature, and the magnitude of temperature-induced volume transition could be modulated by modifying the chromatin compaction state and remodeling processes. Our findings reveal that the cell nucleus can be viewed as a highly charged polymer gel with intriguing thermoresponsive properties, which might play a role in nuclear volume regulation and thermosensing in living cells.

  4. Construction and characterization of two bacterial artificial chromosome libraries of pea (Pisum sativum L.) for the isolation of economically important genes.

    PubMed

    Coyne, C J; McClendon, M T; Walling, J G; Timmerman-Vaughan, G M; Murray, S; Meksem, K; Lightfoot, D A; Shultz, J L; Keller, K E; Martin, R R; Inglis, D A; Rajesh, P N; McPhee, K E; Weeden, N F; Grusak, M A; Li, C-M; Storlie, E W

    2007-09-01

    Pea (Pisum sativum L.) has a genome of about 4 Gb that appears to share conserved synteny with model legumes having genomes of 0.2-0.4 Gb despite extensive intergenic expansion. Pea plant inventory (PI) accession 269818 has been used to introgress genetic diversity into the cultivated germplasm pool. The aim here was to develop pea bacterial artificial chromosome (BAC) libraries that would enable the isolation of genes involved in plant disease resistance or control of economically important traits. The BAC libraries encompassed about 3.2 haploid genome equivalents consisting of partially HindIII-digested DNA fragments with a mean size of 105 kb that were inserted in 1 of 2 vectors. The low-copy oriT-based T-DNA vector (pCLD04541) library contained 55 680 clones. The single-copy oriS-based vector (pIndigoBAC-5) library contained 65 280 clones. Colony hybridization of a universal chloroplast probe indicated that about 1% of clones in the libraries were of chloroplast origin. The presence of about 0.1% empty vectors was inferred by white/blue colony plate counts. The usefulness of the libraries was tested by 2 replicated methods. First, high-density filters were probed with low copy number sequences. Second, BAC plate-pool DNA was used successfully to PCR amplify 7 of 9 published pea resistance gene analogs (RGAs) and several other low copy number pea sequences. Individual BAC clones encoding specific sequences were identified. Therefore, the HindIII BAC libraries of pea, based on germplasm accession PI 269818, will be useful for the isolation of genes underlying disease resistance and other economically important traits.

  5. Lysis gradient centrifugation: a flexible method for the isolation of nuclei from primary cells.

    PubMed

    Katholnig, Karl; Poglitsch, Marko; Hengstschläger, Markus; Weichhart, Thomas

    2015-01-01

    The isolation of nuclei from eukaryotic cells is essential for studying the composition and the dynamic changes of the nuclear proteome to gain insight into the mechanisms of gene expression and cell signalling. Primary cells are particularly challenging for standard nuclear isolation protocols due to low protein content, sample degradation, or nuclear clumping. Here, we describe a rapid and flexible protocol for the isolation of clean and intact nuclei, which results in the recovery of 90-95 % highly pure nuclei. The method, called lysis gradient centrifugation (LGC), is based on an iso-osmolar discontinuous iodixanol-based density gradient including a detergent-containing lysis layer. A single low g-force centrifugation step enables mild cell lysis and prevents extensive contact of the nuclei with the cytoplasmic environment. This fast method shows high reproducibility due to the relatively little cell manipulation required by the investigator. Further advantages are the low amount of starting material required, easy parallel processing of multiple samples, and isolation of nuclei and cytoplasm at the same time from the same sample.

  6. Isolation of liver nuclei that retain functional trans-membrane transport.

    PubMed

    Ho, Y F; Guenthner, T M

    1997-11-01

    We have developed a method for the rapid isolation of hepatocyte nuclei, which employs gentle homogenization and centrifugation conditions, and involves minimal processing time. The purified nuclei were morphologically unaltered when observed by light and electron microscopy. No significant contamination from cytoplasm or mitochondria was detected when assessed by marker enzymes. Membrane transport function, measured as ATP-dependent calcium uptake, was intact. This isolation method was devised to be applicable to studies that involve measurement of uptake and active transport of a variety of substances by the cell nucleus.

  7. DNase I sensitivity of transcriptionally active genes in intact nuclei and isolated chromatin of plants.

    PubMed Central

    Spiker, S; Murray, M G; Thompson, W F

    1983-01-01

    We have investigated the DNase I sensitivity of transcriptionally active DNA sequences in intact nuclei and isolated chromatin from embryos of wheat (Triticum aestivum L.). Nuclei or isolated chromatin was incubated with DNase I, and the extent of DNA digestion was monitored as percentage acid solubility. The resistant DNA and DNA from sham-digested controls were used to drive reassociation reactions with cDNA populations corresponding to either total poly(A)+RNA from unimbibed wheat embryos or polysomal poly(A)+RNA from embryos that had imbibed for 3 hr. Sequences complementary to either probe were depleted in DNase I-resistant DNA from nuclei and from chromatin isolated under low-ionic-strength conditions. This indicates that transcriptionally active sequences are preferentially DNase I sensitive in plants. In chromatin isolated at higher ionic strength, cDNA complementary sequences were not preferentially depleted by DNase I treatment. Therefore, the chromatin structure that confers preferential DNase I sensitivity to transcriptionally active genes appears to be lost when the higher-ionic-strength method of preparation is used. Treatment of wheat nuclei with DNase I causes the release of four prominent nonhistone chromosomal proteins that comigrate with wheat high mobility group proteins on NaDodSO4 gels. Images PMID:6219388

  8. Optimized Methods for the Isolation of Arabidopsis Female Central Cells and Their Nuclei.

    PubMed

    Park, Kyunghyuk; Frost, Jennifer M; Adair, Adam James; Kim, Dong Min; Yun, Hyein; Brooks, Janie S; Fischer, Robert L; Choi, Yeonhee

    2016-10-01

    The Arabidopsis female gametophyte contains seven cells with eight haploid nuclei buried within layers of sporophytic tissue. Following double fertilization, the egg and central cells of the gametophyte develop into the embryo and endosperm of the seed, respectively. The epigenetic status of the central cell has long presented an enigma due both to its inaccessibility, and the fascinating epigenome of the endosperm, thought to have been inherited from the central cell following activity of the DEMETER demethylase enzyme, prior to fertilization. Here, we present for the first time, a method to isolate pure populations of Arabidopsis central cell nuclei. Utilizing a protocol designed to isolate leaf mesophyll protoplasts, we systematically optimized each step in order to efficiently separate central cells from the female gametophyte. We use initial manual pistil dissection followed by the derivation of central cell protoplasts, during which process the central cell emerges from the micropylar pole of the embryo sac. Then, we use a modified version of the Isolation of Nuclei TAgged in specific Cell Types (INTACT) protocol to purify central cell nuclei, resulting in a purity of 75-90% and a yield sufficient to undertake downstream molecular analyses. We find that the process is highly dependent on the health of the original plant tissue used, and the efficiency of protoplasting solution infiltration into the gametophyte. By isolating pure central cell populations, we have enabled elucidation of the physiology of this rare cell type, which in the future will provide novel insights into Arabidopsis reproduction.

  9. Optimized Methods for the Isolation of Arabidopsis Female Central Cells and Their Nuclei

    PubMed Central

    Park, Kyunghyuk; Frost, Jennifer M.; Adair, Adam James; Kim, Dong Min; Yun, Hyein; Brooks, Janie S.; Fischer, Robert L.; Choi, Yeonhee

    2016-01-01

    The Arabidopsis female gametophyte contains seven cells with eight haploid nuclei buried within layers of sporophytic tissue. Following double fertilization, the egg and central cells of the gametophyte develop into the embryo and endosperm of the seed, respectively. The epigenetic status of the central cell has long presented an enigma due both to its inaccessibility, and the fascinating epigenome of the endosperm, thought to have been inherited from the central cell following activity of the DEMETER demethylase enzyme, prior to fertilization. Here, we present for the first time, a method to isolate pure populations of Arabidopsis central cell nuclei. Utilizing a protocol designed to isolate leaf mesophyll protoplasts, we systematically optimized each step in order to efficiently separate central cells from the female gametophyte. We use initial manual pistil dissection followed by the derivation of central cell protoplasts, during which process the central cell emerges from the micropylar pole of the embryo sac. Then, we use a modified version of the Isolation of Nuclei TAgged in specific Cell Types (INTACT) protocol to purify central cell nuclei, resulting in a purity of 75–90% and a yield sufficient to undertake downstream molecular analyses. We find that the process is highly dependent on the health of the original plant tissue used, and the efficiency of protoplasting solution infiltration into the gametophyte. By isolating pure central cell populations, we have enabled elucidation of the physiology of this rare cell type, which in the future will provide novel insights into Arabidopsis reproduction. PMID:27788573

  10. Isolation of Arabidopsis Pollen, Sperm Cells, and Vegetative Nuclei by Fluorescence-Activated Cell Sorting (FACS).

    PubMed

    Santos, Mário R; Bispo, Cláudia; Becker, Jörg D

    2017-01-01

    Efficient methods to isolate highly purified Arabidopsis thaliana pollen and the subcellular components of the male gametophyte (the vegetative nucleus and two sperm cells) have enabled genome-scale studies revealing a highly dynamic reprogramming of the transcriptome and epigenome during pollen development. Here, we describe the isolation of uninucleate microspores, mature pollen, as well as sperm cells and vegetative nuclei by Fluorescence-Activated Cell Sorting.

  11. Protocols for nuclei isolation and nuclear protein extraction from the resurrection plant Xerophyta viscosa for proteomic studies.

    PubMed

    Abdalla, Kamal Omer; Thomson, Jennifer Ann; Rafudeen, Muhammad Suhail

    2009-01-15

    The plant nucleus is an important subcellular organelle but the isolation of pure and enriched nuclei from plants and subsequent extraction of nuclear proteins for proteomic studies is challenging. Here, we present protocols for nuclei isolation and nuclear protein extraction from the resurrection plant, Xerophyta viscosa, and show optimization and modification of the most critical steps.

  12. Isolation and characterization of NADP+-linked isocitrate dehydrogenase of germinating pea seeds (Pisum sativum).

    PubMed

    Srivastava, P K; Singh, D S

    2001-10-01

    NADP+-linked isocitrate dehydrogenase (E.C.1.1.1.42) has been purified to homogeneity from germinating pea seeds. The enzyme is a tetrameric protein (mol wt, about 146,000) made up of apparently identical monomers (subunit mol wt, about 36,000). Thermal inactivation of purified enzyme at 45 degrees and 50 degrees C shows simple first order kinetics. The enzyme shows optimum activity at pH range 7.5-8. Effect of substrate [S] on enzyme activity at different pH (6.5-8) suggests that the proton behaves formally as an "uncompetitive inhibitor". A basic group of the enzyme (site) is protonated in this pH range in the presence of substrate only, with a pKa equal to 6.78. On successive dialysis against EDTA and phosphate buffer, pH 7.8 at 0 degrees C, yields an enzymatically inactive protein showing kinetics of thermal inactivation identical to the untreated (native) enzyme. Maximum enzyme activity is observed in presence of Mn2+ and Mg2+ ions (3.75 mM). Addition of Zn2+, Cd2+, Co2+ and Ca2+ ions brings about partial recovery. Other metal ions Fe2+, Cu2+ and Ni2+ are ineffective.

  13. Iron-induced DNA damage and synthesis in isolated rat liver nuclei.

    PubMed Central

    Shires, T K

    1982-01-01

    Incubation of iron with isolated rat liver nuclei stimulated fragmentation of single-stranded DNA, incorporation of [3H]thymidine into DNA and the binding of 59Fe to DNA. FeCl2 was about twice as active as FeCl3. Lipid peroxidation took place in nuclei incubated with FeCl2, but not with FeCl3. Generation of reactive forms of oxygen was required for iron-mediated DNA damage, but evidence for direct interaction of reactive oxygen with DNA was not found. Apparent adducts of iron bound to DNA seemed to be formed by an enzymic mechanism. PMID:7138506

  14. RNA synthesis in isolated brian nuclei after administration of d-lysergic acid diethylamide (LSD) in vivo.

    PubMed Central

    Brown, I R

    1975-01-01

    RNA synthesis in isolated brain nuclei was analyzed 2.5 hr after the intravenous administration of d-lysergic acid diethylamide (LSD) to young rabbits. The drug stimulated transcription by 54% in brain stem nuclei and by 13% in cerebral hemisphere nuclei expressed over saline controls. Both nucleoplasmic and nucleolar RNA synthesis were increased. The main activity in the isolated nuclei assay was due to nucleoplasmic RNA polymerase, since alpha-amanitin reduced synthesis by over 70% in either drug or control treatments. PMID:1055380

  15. Evaluation of the participation of ferredoxin in oxygen reduction in the photosynthetic electron transport chain of isolated pea thylakoids.

    PubMed

    Kozuleva, Marina A; Ivanov, Boris N

    2010-07-01

    The contribution to reduction of oxygen by ferredoxin (Fd) to the overall reduction of oxygen in isolated pea thylakoids was studied in the presence of Fd versus Fd + NADP(+). The overall rate of electron transport was measured using a determination of Photosystem II quantum yield from chlorophyll fluorescence parameters, and the rate of oxidation of Fd was measured from the light-induced redox changes of Fd. At low light intensity, increasing Fd concentration from 5 to 30 microM in the absence of NADP(+) increased the proportion of oxygen reduction by Fd from 25-35 to 40-60% in different experiments. This proportion decreased with increasing light intensity. When NADP(+) was added in the presence of 15 microM Fd, which was optimal for the NADP(+) reduction rate, the participation of Fd in the reduction of oxygen was low, no more than 10%, and it also decreased with increasing light intensity. At high light intensity, the overall oxygen reduction rates in the presence of Fd + NADP(+) and in the presence of Fd alone were comparable. The significance of reduction of dioxygen either by water-soluble Fd or by the membrane-bound carriers of the photosynthetic electron transport chain for redox signaling under different light intensities is discussed.

  16. Identification of a new pea gene, PsNlec1, encoding a lectin-like glycoprotein isolated from the symbiosomes of root nodules.

    PubMed Central

    Kardailsky, I V; Sherrier, D J; Brewin, N J

    1996-01-01

    A 27-kD glycoprotein antigen recognized by monoclonal antibody MAC266 was purified from isolated symbiosomes derived from pea (Pisum sativum) root nodules containing Rhizobium. The N-terminal amino acid sequence was obtained, and the corresponding cDNA clone was isolated by a polymerase chain reaction-based strategy. The clone contained a single open reading frame, and the gene was termed PsNlec1. Phylogenetic analysis of 31 legume sequences showed that the PsNlec1 protein is related to the legume lectin family but belongs to a subgroup that is very different from pea seed lectin. Expression of the PsNlec1 transcript was much stronger in nodules than in other parts of the plant. It was found in both infected and uninfected cells in the central tissue of the nodule and in the stele of the root near the attachment point of the nodule. When uninfected pea seedlings were grown on medium containing nitrate, weak transcription of PsNlec1 was observed in the root system. The identification of PsNlec1 inside the symbiosome is consistent with the observation that legume lectins are generally vacuolar proteins that may serve as transient storage components. PMID:8685275

  17. Isolation of cell nuclei using inert macromolecules to mimic the crowded cytoplasm.

    PubMed

    Hancock, Ronald; Hadj-Sahraoui, Yasmina

    2009-10-23

    Cell nuclei are commonly isolated and studied in media which include millimolar concentrations of cations, which conserve the nuclear volume by screening the negative charges on chromatin and maintaining its compaction. However, two factors question if these ionic conditions correctly reproduce the environment of nuclei in vivo: the small-scale motion and conformation of chromatin in vivo are not reproduced in isolated nuclei, and experiments and theory suggest that small ions in the cytoplasm are not free in the soluble phase but are predominantly bound to macromolecules. We studied the possible role in maintaining the structure and functions of nuclei in vivo of a further but frequently overlooked property of the cytoplasm, the crowding or osmotic effects caused by diffusible macromolecules whose concentration, measured in several studies, is in the range of 130 mg/ml. Nuclei which conserved their volume in the cell and their ultrastructure seen by electron microscopy were released from K562 cells in media containing the inert polymer 70 kDa Ficoll (50% w/v) or 70 kDa dextran (35% w/v) to replace the diffusible cytoplasmic molecules which were dispersed on cell lysis with digitonin, with 100 microM K-Hepes buffer as the only source of ions. Immunofluorescence labelling and experiments using cells expressing GFP-fusion proteins showed that internal compartments (nucleoli, PML and coiled bodies, foci of RNA polymerase II) were conserved in these nuclei, and nascent RNA transcripts could be elongated. Our observations are consistent with the hypothesis that crowding by diffusible cytoplasmic macromolecules is a crucial but overlooked factor which supports the nucleus in vivo by equilibrating the opposing osmotic pressure cause by the high concentration of macromolecules in the nucleus, and suggest that crowded media provide more physiological conditions to study nuclear structure and functions. They may also help to resolve the long-standing paradox that the small

  18. Iron absorption from experimental infant formulas based on pea (Pisum sativum)-protein isolate: the effect of phytic acid and ascorbic acid.

    PubMed

    Davidsson, L; Dimitriou, T; Walczyk, T; Hurrell, R F

    2001-01-01

    Infant formula based on pea (Pisum sativum)-protein isolate has been suggested as an alternative to soybean formula in countries where soybean is not a native crop, or when soybean protein cannot be used due to allergic reactions or intolerances. In the present study, Fe absorption from experimental infant formulas based on pea-protein isolate was measured in healthy non-anaemic young women. The influence of phytic acid and ascorbic acid on Fe absorption was evaluated, using a stable-isotope technique based on incorporation of Fe stable-isotope labels into erythrocytes 14 d after administration. Geometric mean Fe absorption increased from 20.7 (+1 SD 41.6, -1 SD 10.3) % to 33.1 (+1 SD 58.6, -1 SD 18.7) %; (P < 0.0001; n 10) after enzymic degradation of virtually all phytic acid. Doubling the molar ratio Fe:ascorbic acid from 1:2.1 to 1:4.2 in the infant formula with native phytic acid content also increased Fe absorption significantly (P < 0.0001; n 10); geometric mean Fe absorption increased from 14.8 (+1 SD 32.1, -1 SD 6.8) % to 22.1 (+1 SD 47.2, -1 SD 10.4) %. These results confirm the inhibitory and enhancing effects of phytic acid and ascorbic acid respectively on Fe absorption, but also indicate relatively high fractional Fe absorption from the pea-protein-based formulas. After adjusting for differences in Fe status, our data indicate that Fe absorption from dephytinised pea protein might be less inhibitory than dephytinised soybean protein as measured in a previous study (Hurrell et al. 1998).

  19. Power-law rheology of isolated nuclei with deformation mapping of nuclear substructures.

    PubMed

    Dahl, Kris Noel; Engler, Adam J; Pajerowski, J David; Discher, Dennis E

    2005-10-01

    Force-induced changes in genome expression as well as remodeling of nuclear architecture in development and disease motivate a deeper understanding of nuclear mechanics. Chromatin and green fluorescent protein-lamin B dynamics were visualized in a micropipette aspiration of isolated nuclei, and both were shown to contribute to viscoelastic properties of the somatic cell nucleus. Reversible swelling by almost 200% in volume, with changes in salt, demonstrates the resilience and large dilational capacity of the nuclear envelope, nucleoli, and chromatin. Swelling also proves an effective way to separate the mechanical contributions of nuclear elements. In unswollen nuclei, chromatin is a primary force-bearing element, whereas swollen nuclei are an order of magnitude softer, with the lamina sustaining much of the load. In both cases, nuclear deformability increases with time, scaling as a power law-thus lacking any characteristic timescale-when nuclei are either aspirated or indented by atomic force microscopy. The nucleus is stiff and resists distortion at short times, but it softens and deforms more readily at longer times. Such results indicate an essentially infinite spectrum of timescales for structural reorganization, with implications for regulating genome expression kinetics.

  20. Power-Law Rheology of Isolated Nuclei with Deformation Mapping of Nuclear Substructures

    PubMed Central

    Dahl, Kris Noel; Engler, Adam J.; Pajerowski, J. David; Discher, Dennis E.

    2005-01-01

    Force-induced changes in genome expression as well as remodeling of nuclear architecture in development and disease motivate a deeper understanding of nuclear mechanics. Chromatin and green fluorescent protein-lamin B dynamics were visualized in a micropipette aspiration of isolated nuclei, and both were shown to contribute to viscoelastic properties of the somatic cell nucleus. Reversible swelling by almost 200% in volume, with changes in salt, demonstrates the resilience and large dilational capacity of the nuclear envelope, nucleoli, and chromatin. Swelling also proves an effective way to separate the mechanical contributions of nuclear elements. In unswollen nuclei, chromatin is a primary force-bearing element, whereas swollen nuclei are an order of magnitude softer, with the lamina sustaining much of the load. In both cases, nuclear deformability increases with time, scaling as a power law—thus lacking any characteristic timescale—when nuclei are either aspirated or indented by atomic force microscopy. The nucleus is stiff and resists distortion at short times, but it softens and deforms more readily at longer times. Such results indicate an essentially infinite spectrum of timescales for structural reorganization, with implications for regulating genome expression kinetics. PMID:16055543

  1. Mercurated nucleotides: assessment of a new tool to study RNA synthesis and processing in isolated nuclei.

    PubMed Central

    Schäfer, K P

    1977-01-01

    Mercurated pyrimidine nucleotides have been used to study RNA synthesis and processing in isolated nuclei from mouse L cells. 5-mercuridine triphosphate (5-Hg-UTP) or 5-Hg-CTP are accepted as substrates by the purified RNA polymerases (I+III) and (II) from mouse cells, respectively, as well as by the enzymes still bound to the nuclear chromatin. In nuclei, RNA synthesis in the presence of Hg-UTP is reduced to 60-70% of a control. 30-60% of RNA labeled in vitro with (3H)UTP in isolated nuclei is not retained on sulfhydryl sepharose columns. Sucrose gradient analysis reveals a size distribution of the non-bound RNA similar to non-mercurated control RNA. Hg-RNA is found in a single peak from 4-10S. Chase experiments indicate that this RNA is the original transcript. It is argued that Hg-nucleotides may cause premature chain termination. Methylation of RNA in vitro by S-adenosyl methionine ((3H)SAM) is reduced to 75% of controls in the presence of Hg-UTP. Only 6% of the methyl groups appear in Hg-RNA. Polyadenylation is reduced as well. 15% of poly(A) (+)RNA are found in control assays whereas only 1% of Hg-RNA carries a poly(A) end added in vitro. These results limit the use of mercurated nucleotides for studies of nuclear RNA synthesis and processing. PMID:600804

  2. RNA metabolism in isolated nuclei: processing and transport of immunoglobulin light chain sequences.

    PubMed Central

    Otegui, C; Patterson, R J

    1981-01-01

    Transport of prelabeled RNA from isolated myeloma nuclei is studied using conditions that permit RNA synthesis. Cytosol and spermidine are not required to maintain nuclear stability and inhibited RNA release. Omission of ATP or GTP decreased release 25 to 40%. The stimulatory effect of ATP or GTP is not due to hydrolysis of the triphosphates by the nuclear envelope NTPase, since addition of quercetin (an inhibitor of this NTPase) has no effect on the quantity of RNA released. The size distribution and percentage of poly A-containing species released from nuclei incubated with or without ATP or the other rNTPs are identical. Hybridization analysis of nuclear RNA before the transport assay revealed mature and precursor k light chain mRNA sequences. Following the transport assay, a significant fraction of k mRNA precursors is chased into mature k mRNA which is found both in nuclear-retained and released RNA. PMID:6795596

  3. A novel lipid transfer protein from the pea Pisum sativum: isolation, recombinant expression, solution structure, antifungal activity, lipid binding, and allergenic properties.

    PubMed

    Bogdanov, Ivan V; Shenkarev, Zakhar O; Finkina, Ekaterina I; Melnikova, Daria N; Rumynskiy, Eugene I; Arseniev, Alexander S; Ovchinnikova, Tatiana V

    2016-04-30

    Plant lipid transfer proteins (LTPs) assemble a family of small (7-9 kDa) ubiquitous cationic proteins with an ability to bind and transport lipids as well as participate in various physiological processes including defense against phytopathogens. They also form one of the most clinically relevant classes of plant allergens. Nothing is known to date about correlation between lipid-binding and IgE-binding properties of LTPs. The garden pea Pisum sativum is widely consumed crop and important allergenic specie of the legume family. This work is aimed at isolation of a novel LTP from pea seeds and characterization of its structural, functional, and allergenic properties. Three novel lipid transfer proteins, designated as Ps-LTP1-3, were found in the garden pea Pisum sativum, their cDNA sequences were determined, and mRNA expression levels of all the three proteins were measured at different pea organs. Ps-LTP1 was isolated for the first time from the pea seeds, and its complete amino acid sequence was determined. The protein exhibits antifungal activity and is a membrane-active compound that causes a leakage from artificial liposomes. The protein binds various lipids including bioactive jasmonic acid. Spatial structure of the recombinant uniformly (13)C,(15)N-labelled Ps-LTP1 was solved by heteronuclear NMR spectroscopy. In solution the unliganded protein represents the mixture of two conformers (relative populations ~ 85:15) which are interconnected by exchange process with characteristic time ~ 100 ms. Hydrophobic residues of major conformer form a relatively large internal tunnel-like lipid-binding cavity (van der Waals volume comes up to ~1000 Å(3)). The minor conformer probably corresponds to the protein with the partially collapsed internal cavity. For the first time conformational heterogeneity in solution was shown for an unliganded plant lipid transfer protein. Heat denaturation profile and simulated gastrointestinal digestion assay showed that Ps

  4. AN OPTIMIZED METHOD FOR THE ISOLATION OF NUCLEI FROM CHLAMYDOMONAS REINHARDTII (CHLOROPHYCEAE)(1).

    PubMed

    Winck, Flavia Vischi; Kwasniewski, Miroslaw; Wienkoop, Stefanie; Mueller-Roeber, Bernd

    2011-04-01

    The cell nucleus harbors a large number of proteins involved in transcription, RNA processing, chromatin remodeling, nuclear signaling, and ribosome assembly. The nuclear genome of the model alga Chlamydomonas reinhardtii P. A. Dang. was recently sequenced, and many genes encoding nuclear proteins, including transcription factors and transcription regulators, have been identified through computational discovery tools. However, elucidating the specific biological roles of nuclear proteins will require support from biochemical and proteomics data. Cellular preparations with enriched nuclei are important to assist in such analyses. Here, we describe a simple protocol for the isolation of nuclei from Chlamydomonas, based on a commercially available kit. The modifications done in the original protocol mainly include alterations of the differential centrifugation parameters and detergent-based cell lysis. The nuclei-enriched fractions obtained with the optimized protocol show low contamination with mitochondrial and plastid proteins. The protocol can be concluded within only 3 h, and the proteins extracted can be used for gel-based and non-gel-based proteomic approaches. © 2011 Phycological Society of America.

  5. Automated segmentation and isolation of touching cell nuclei in cytopathology smear images of pleural effusion using distance transform watershed method

    NASA Astrophysics Data System (ADS)

    Win, Khin Yadanar; Choomchuay, Somsak; Hamamoto, Kazuhiko

    2017-06-01

    The automated segmentation of cell nuclei is an essential stage in the quantitative image analysis of cell nuclei extracted from smear cytology images of pleural fluid. Cell nuclei can indicate cancer as the characteristics of cell nuclei are associated with cells proliferation and malignancy in term of size, shape and the stained color. Nevertheless, automatic nuclei segmentation has remained challenging due to the artifacts caused by slide preparation, nuclei heterogeneity such as the poor contrast, inconsistent stained color, the cells variation, and cells overlapping. In this paper, we proposed a watershed-based method that is capable to segment the nuclei of the variety of cells from cytology pleural fluid smear images. Firstly, the original image is preprocessed by converting into the grayscale image and enhancing by adjusting and equalizing the intensity using histogram equalization. Next, the cell nuclei are segmented using OTSU thresholding as the binary image. The undesirable artifacts are eliminated using morphological operations. Finally, the distance transform based watershed method is applied to isolate the touching and overlapping cell nuclei. The proposed method is tested with 25 Papanicolaou (Pap) stained pleural fluid images. The accuracy of our proposed method is 92%. The method is relatively simple, and the results are very promising.

  6. Isolation of PsPIN2 and PsAUX1 from etiolated pea epicotyls and their expression on a three-dimensional clinostat

    NASA Astrophysics Data System (ADS)

    Hoshino, Tomoki; Hitotsubashi, Reiko; Miyamoto, Kensuke; Tanimoto, Eiichi; Ueda, Junichi

    We isolated novel cDNAs containing the complete open reading frames of a putative auxin influx carrier, PsAUX1, and a putative auxin efflux carrier, PsPIN2, from etiolated pea epicotyls. High levels of homology were found on nucleotide and deduced amino acid sequences among PsPIN2, PsPIN1 (Accession No. AY222857) and AtPINs. Phylogenetic analyses based on deduced amino acid sequences revealed that PsPIN2 belonged to a subclade including AtPIN3, AtPIN4 and AtPIN7, while PsPIN1 belonged to the same clade as AtPIN1. The results were similar for PsAUX1 and AtAUX1, where PsAUX1 belongs to the same subclade as AtAUX1 and CS-AUX1. Expression of PsPIN1, PsPIN2 and PsAUX1 in pea epicotyl segments was promoted upon incubation of the segments with auxin (indole-3-acetic acid). In 3.5-d-old etiolated pea seedlings, relatively high expression of PsPIN1 and PsAUX1 was observed in the hook region, growing epicotyls and root tips as compared with those in mature regions of epicotyls and roots. Expression of PsPIN2 in roots was less than that in shoots. Simulated microgravity conditions on a three-dimensional clinostat remarkably increased gene expression of PsPIN1 and PsAUX1 in the hook and the internodes of pea epicotyls, but the increase in PsPIN2 was less. In contrast, polar auxin transport of pea epicotyls was substantially suppressed under simulated microgravity conditions on a 3D clinostat, similar to data from a space experiment on STS-95. These results suggest that PsPINs and PsAUX1 are auxin-inducible genes, and that the expression of PsPINs and PsAUX1 genes is sensitive to gravistimulation.

  7. Rapid isolation of nuclei from living immune cells by a single centrifugation through a multifunctional lysis gradient.

    PubMed

    Poglitsch, Marko; Katholnig, Karl; Säemann, Marcus D; Weichhart, Thomas

    2011-10-28

    Due to their low protein content and limited nuclear detergent stability, primary human immune cells such as monocytes or T lymphocytes represent a great challenge for standard nuclear isolation protocols. Nuclei clumping during the multiple centrifugation steps or contamination of isolated nuclei with cytoplasmic proteins due to membrane lysis is a frequently observed problem. Here we describe a versatile and novel method for the isolation of clean and intact nuclei from primary human monocytes, which can be applied for virtually any cell type. Living cells were applied on an iso-osmolar discontinuous iodixanol-based density gradient including a detergent-containing lysis layer. Mild cell lysis as well as efficient washing of the nuclei was performed during the course of one single low g-force centrifugation step. The isolation procedure, which we call lysis gradient centrifugation (LGC), results in the recovery of 90-95% of highly pure nuclei. This easy and highly reproducible procedure allows an effective preparation of nuclei and the cytoplasm in only 15 min with the ability to handle as little as one million cells per sample and easy parallel processing of multiple samples.

  8. [Isolation of various fractions of human placental nuclei and their utilization in an in vitro transcription system].

    PubMed

    Lasbennes, A; Hess, K; Ramboarina, L; Belleville, F; Nabet, P; Paysant, P

    1981-01-01

    A new approach to isolating chromatin and DNA from human placental nuclei is described. Criteria of purity are given by spectrophotometry and chemical analysis. Chromatin and DNA keep their template efficiencies after isolation. We used them in a transcription system where wheat germ RNA polymerase intervenes. The role of histones was tested on this system.

  9. Isolated nuclei adapt to force and reveal a mechanotransduction pathway in the nucleus.

    PubMed

    Guilluy, Christophe; Osborne, Lukas D; Van Landeghem, Laurianne; Sharek, Lisa; Superfine, Richard; Garcia-Mata, Rafael; Burridge, Keith

    2014-04-01

    Mechanical forces influence many aspects of cell behaviour. Forces are detected and transduced into biochemical signals by force-bearing molecular elements located at the cell surface, in adhesion complexes or in cytoskeletal structures. The nucleus is physically connected to the cell surface through the cytoskeleton and the linker of nucleoskeleton and cytoskeleton (LINC) complex, allowing rapid mechanical stress transmission from adhesions to the nucleus. Although it has been demonstrated that nuclei experience force, the direct effect of force on the nucleus is not known. Here we show that isolated nuclei are able to respond to force by adjusting their stiffness to resist the applied tension. Using magnetic tweezers, we found that applying force on nesprin-1 triggers nuclear stiffening that does not involve chromatin or nuclear actin, but requires an intact nuclear lamina and emerin, a protein of the inner nuclear membrane. Emerin becomes tyrosine phosphorylated in response to force and mediates the nuclear mechanical response to tension. Our results demonstrate that mechanotransduction is not restricted to cell surface receptors and adhesions but can occur in the nucleus.

  10. Aphidicolin inhibits DNA synthesis by DNA polymerase alpha and isolated nuclei by a similar mechanism.

    PubMed Central

    Krokan, H; Wist, E; Krokan, R H

    1981-01-01

    Aphidicolin is a selective inhibitor of DNA polymerase alpha. In contrast to earlier reports, the drug was found to inhibit DNA synthesis catalyzed by DNA polymerase alpha and isolated HeLa cell nuclei by a similar mechanism. For both systems aphidicolin primarily competed with dCTP incorporation. However, the apparent Vmax for dCTP incorporation was reduced by 50-60% at relatively low concentrations of aphidicolin, thus the mechanism of inhibition is complex. Furthermore, a 2-5 fold increase in apparent Km for dTTP was observed in the presence of aphidicolin, but the apparent Km values for dATP and dGTP were essentially unaltered. This, together with additional evidence, suggested that the mechanism of action of aphidicolin involves a strong competition with dCMP incorporation, a weaker competition with dTMP incorporation and very little, if any, competition with dGMP and dAMP incorporation. PMID:6795595

  11. Effects of Arsenite, Sulfite, and Sulfate on Photosynthetic Carbon Metabolism in Isolated Pea (Pisum sativum L., cv Little Marvel) Chloroplasts 1

    PubMed Central

    Marques, Ivano A.; Anderson, Louise E.

    1986-01-01

    Photosynthetic CO2-fixation in isolated pea (Pisum sativum L., cv Little Marvel) chloroplasts during induction is markedly inhibited by 0.4 millimolar sulfite. Sulfate at the same concentration has almost no effect. The 14CO2-fixation pattern indicates that the primary effect of sulfite is inhibition of the reaction catalyzed by ribulose bisphosphate carboxylase and a stimulation of export of intermediates out of the chloroplasts. Inhibition of light modulation of stromal enzyme activity does not appear to account for the toxicity of SO2 in this Pisum variety. Arsenite at 0.2 millimolar concentrations inhibits light activation and inhibits photosynthetic CO2 fixation. The 14CO2-fixation pattern indicates that the primary effect of arsenite is inhibition of light activation of reductive pentose phosphate pathway enzyme activity. PMID:16665056

  12. Isolation and in silico analysis of promoter of a high salinity stress-regulated pea DNA helicase 45.

    PubMed

    Tajrishi, Marjan M; Tuteja, Narendra

    2011-10-01

    Helicases are motor proteins that can transiently catalyze the unwinding of energetically stable duplex DNA or RNA molecules by using ATP hydrolysis as the source of energy. Many helicases share a core region of highly conserved sequence motifs, and belong to the rapidly growing DEAD-box protein family. Pea DNA helicase 45 (PDH45), that exhibits striking homology with eukaryotic translation initiation factor 4A (eIF4A), contains ATP-dependent DNA and RNA helicase, DNA-dependent ATPase, and ATP-binding activities. The transcript of the PDH45 gene was reported to be upregulated in pea plant in response to high salinity, cold stress, abscisic acid (ABA), dehydration and early wounding. The first direct evidence that overexpression of PDH45 confers salinity stress tolerance without yield loss has also been reported. A promoter analysis of PDH45 gene has not been studied. The cis-regulatory elements present on promoter region of the gene act as binding sites for RNA polymerase and transcription factors and control the regulation of gene expression. Here we report the promoter of the PDH45 gene that contains stress-responsive cis-regulatory elements which may be responsible for regulating the expression of PDH45 under abiotic stress conditions.

  13. Pea Xyloglucan and Cellulose 1

    PubMed Central

    Hayashi, Takahisa; Polonenko, Daniel R.; Camirand, Anne; Maclachlan, Gordon

    1986-01-01

    The synthesis and assembly of xyloglucan were examined during early stages of wall regeneration by protoplasts isolated from growing regions of etiolated peas. During early stages of cultivation, fluorescence microscopy showed that the protoplast surface bound Calcofluor and ammonium salt of 8-anilino-1-naphthalene sulfonic acid and, in time, it also bound fluorescent fucose-binding lectin. Based on chemical analysis, 1,3-β-glucan was the main polysaccharide formed by protoplasts and xyloglucan and cellulose were minor wall components. Binding between cellulose and xyloglucan was not as strong as that in tissues of intact pea plants, i.e. mild alkali could dissolve most xyloglucan from the protoplast. However, the addition of exogenous pea xyloglucan into the culture medium stimulated the deposition of new polysaccharides into the protoplast wall and enhanced the close association of newly formed xyloglucan with cellulose. Images Fig. 1 Fig. 4 PMID:16665011

  14. Characterization of a DNA uptake reaction through the nuclear membrane of isolated yeast nuclei. [Saccharomyces cerevisiae

    SciTech Connect

    Tsuchiya, E.; Shakuto, S.; Miyakawa, T.; Fukui, S.

    1988-02-01

    Isolated yeast nuclei were able to incorporate /sup 3/H-labeled pJDB219 DNA in vitro in the presence of ATP and Mg/sup 2 +/. The number of plasmid molecules incorporated into each nucleus was calculated to be 60 under the conditions we used. Enzyme-histochemical staining of the incorporated biotinylated pJDB219 with streptavidin-biotinylated-peroxidase complex indicated a uniform distribution of the incorporated plasmids within each nucleus. After intranuclear incorporation, substrate pJDB219 DNAs (open and closed circular forms) were changed to the linear form and were weakly digested over the longer incubation period (over 60 min). Facile release of the once-incorporated plasmid DNA was never observable; discharge of the incorporated (/sup 3/H)pJDB219 during a 60-min incubation was less than 5%. The addition of adenylyl-imidodiphosphate, N,N'-dicyclohexylcarbodiimide (DCCD), or quercetin inhibited in vitro DNA uptake reaction. DCCD and quercetin inhibited the nuclear ATPase and apparent protein kinase, respectively; hence, the involvement of these enzymes in the nuclear DNA transport system was suggested.

  15. Isolation of cell nuclei in microchannels by short-term chemical treatment via two-step carrier medium exchange.

    PubMed

    Toyama, Kaori; Yamada, Masumi; Seki, Minoru

    2012-08-01

    Separation/purification of nuclei from cells is a critical process required for medical and biochemical research applications. Here, we report a flow-through microfluidic device for isolating cell nuclei by selectively digesting the cell membrane by using the concept of hydrodynamic filtration (HDF). When a cell suspension is continuously introduced into a microchannel (main channel) possessing multiple side channels, cells flow through the main channel, whereas the carrier medium of the cells is drained through the side channels. Introductions of a cell treatment solution containing a surfactant and a washing buffer enable the two-step exchange of the carrier-medium and the cell treatment by the surfactant for a short span of time. The precise control of the treatment time by changing the flow rate and/or the size of the microchannel enables the selective digestion of cell membranes, resulting in the isolation of cell nuclei after separation from membrane debris and cytoplasmic components according to size. We examined several surfactant molecules and demonstrated that Triton X-100 exhibited high efficiency regarding nucleus isolation for both adherent (HeLa) and nonadherent (JM) cells, with a recovery ratio of ~80 %. In addition, the isolation efficiency was evaluated by western blotting. The presented flow-through microfluidic cell-nucleus separator may be a useful tool for general biological applications, because of its simplicity in operation, high reproducibility, and accuracy.

  16. Ultrastructural and biochemical observations on interphase nuclei isolated from chicken erythrocytes.

    PubMed

    Walmsley, M E; Davies, H G

    1975-01-01

    Adult hen erythrocyte nuclei are isolated from cells or haemolysed in situ by acting on the plasma membrane with rotating knives or with non-ionic detergents. When the isolation medium contains magnesium ions (1 mM), sucrose (0-4 M) and Tris buffer (0.01 M, pH 7-5) called SMTOG (see text), the ultrastructure in thin sections through the condensed chromatin bodies, after staining with either uranyl-lead or phosphotungstic acid (PTA), is similar to that found in the intact cell. Hence it can be concluded that the 2 phases which comprise chromatin, the o- and e-phase, survive nuclear isolation. These are so called because the structural units in chromatin are arranged at the surface of the nucleus into one or more layers and give rise to oddly (o) and evenly (e) numbered bands. The 0-phase is also largely retained after extensive washing in 0-07 M NaC1 as shown by electron microscopy and biochemical measurements; only 6% of the total nuclear protein is removed, a value small compared with the fractional amount of the chromatin protein calculated to lie in the o-phase, about 70%. After extensive washing in saline-EDTA there are structural changes in chromatin, but biochemical data show that the molecules in the o-phase are also largely retained; loss of protein amounts to between 5 and 11%. These data suggest that the o-phase is a structural component of the chromatin bodies. They support the hypothesis that condensed chromatin is formed by folding superunit threads. These units consist of a central thread-like element about 17 nm diameter which stains preferentially with uranyl-lead and forms the e-phase, with an outer cylindrical shell forming the o-phase of total diameter about 28nm. The 5-10% proteins removed by salt washes are located exclusively in a particulate component, quite likely the chromatin. They have been examined by sodium dodecyl sulphate (SDS) polyacrylamide gel electrophoresis. There are about 10 or more protein species, ranging in molecular weight

  17. The effects of copper and other ions on the ribonucleic acid polymerase activity of isolated rat liver nuclei

    PubMed Central

    Novello, F.; Stirpe, F.

    1969-01-01

    1. The effects of various ions on the Mg2+- and Mn2+/ammonium sulphate-activated RNA polymerase activities of isolated liver nuclei were studied. 2. The Mg2+-activated RNA polymerase reaction was inhibited by more than 60% by Cd2+, SeO32−, Be2+, Cu2+, Co2+, Ca2+ and La3+, all at 1mm concentrations. 3. The Mn2+/ammonium sulphate-activated RNA polymerase reaction was strongly inhibited by Hg2+, Cd2+, Cu2+ and Ag+. The effect of Hg2+, Cd2+ and Ag+ was relieved by cysteine or mercaptoethanol. 4. Inhibition by Cu2+ was not affected by addition of DNA, and was relieved only partially by EDTA or histidine. 5. No changes of RNA polymerase activities were observed in nuclei isolated from the liver of rats treated with copper albuminate. PMID:4975630

  18. A cell line with decreased sensitivity to the methyl mercury-induced stimulation of alpha-amanitin sensitive RNA synthesis in isolated nuclei.

    PubMed

    Frenkel, G D; Ducote, J; Reboulleau, C P; Gierthy, J

    1988-01-01

    1. In nuclei isolated from cells of the B50 rat neuroblastoma line the stimulatory effect of methyl mercury on alpha-amanitin-sensitive RNA synthesis is very much reduced compared to the stimulatory effect in HeLa nuclei (see: Frenkel G. D. and Randles K. (1982) Specific stimulation of alpha-amanitin-sensitive RNA synthesis in isolated HeLa nuclei by methyl mercury. J. biol. Chem. 257, 6275-6279). 2. The stimulatory effect of another mercury compound, p-hydroxymercuribenzoate, was also much less pronounced in the B50 nuclei. 3. Similar results were obtained with nuclei isolated from B50 cells which had been induced to differentiate by exposure to dibutaryl cyclic AMP. 4. Nuclei isolated from cells of another rat neuroblastoma line (B35), and nuclei from cells of a human neuroblastoma line both exhibited levels of stimulation similar to that of HeLa nuclei. 5. The B50 and HeLa cells were also compared as to their sensitivity to other effects of methyl mercury.

  19. Production of Laccase by a New Myrothecium verrucaria MD-R-16 Isolated from Pigeon Pea [Cajanus cajan (L.) Millsp.] and its Application on Dye Decolorization.

    PubMed

    Sun, Jiao; Guo, Na; Niu, Li-Li; Wang, Qing-Fang; Zang, Yu-Ping; Zu, Yuan-Gang; Fu, Yu-Jie

    2017-04-23

    The present study was conducted to screen a laccase-producing fungal endophyte, optimize fermentation conditions, and evaluate the decolorization ability of the laccase. A new fungal endophyte capable of laccase-producing was firstly isolated from pigeon pea and identified as Myrothecium verrucaria based on a ITS-rRNA sequences analysis. Meanwhile, various fermentation parameters on the laccase production were optimized via response surface methodology (RSM). The optimal fermentation conditions were a fermentation time of five days, temperature 30 °C and pH 6.22. Laccase activity reached 16.52 ± 0.18 U/mL under the above conditions. Furthermore, the laccase showed effective decolorization capability toward synthetic dyes (Congo red, Methyl orange, Methyl red, and Crystal violet) in the presence of the redox mediator ABTS, with more than 70% of dyes decolorizing after 24 h of incubation. Additionally, the activity of laccase was relatively stable with pH (4.5-6.5) and a temperature range of 35-55 °C. Therefore, the high laccase production of the strain and the new fungal laccase could provide a promising alterative approach for industrial and environmental applications.

  20. Transport of amino acids (L-valine, L-lysine, L-glutamic acid) and sucrose into plasma membrane vesicles isolated from cotyledons of developing pea seeds.

    PubMed

    de Jong, A; Borstlap, A C

    2000-10-01

    Transport of the amino acids L-valine, L-lysine, and L-glutamic acid and of sucrose was studied in plasma membrane vesicles isolated from developing cotyledons of pea (Pisum sativum L. cv. Marzia). The vesicles were obtained by aqueous polymer two-phase partitioning of a microsomal fraction and the uptake was determined after the imposition of a H(+)-gradient (DeltapH, inside alkaline) and/or an electrical gradient (Deltapsi, inside negative) across the vesicle membrane. In the absence of gradients, a distinct, time-dependent uptake of L-valine was measured, which could be enhanced about 2-fold by the imposition of DeltapH. The imposition of Deltapsi stimulated the influx of valine by 20%, both in the absence and in the presence of DeltapH. Uptake of L-lysine was more strongly stimulated by Deltapsi than by DeltapH, and its DeltapH-dependent uptake was enhanced about 6-fold by the simultaneous imposition of Deltapsi. In the absence of gradients the uptake of L-glutamic acid was about 2-fold higher than that of L-valine, but it was not detectably affected by DeltapH or Deltapsi. Although the transport of sucrose was very low, a stimulating effect of DeltapH could be clearly demonstrated. The results lend further support to the contention that during seed development cotyledonary cells employ H(+)-symporters for the active uptake of sucrose and amino acids.

  1. Quality improvement of a rice-substituted fried noodle by utilizing the protein-polyphenol interaction between a pea protein isolate and green tea (Camellia sinensis) extract.

    PubMed

    Song, Youngwoon; Yoo, Sang-Ho

    2017-11-15

    The quality of rice-substituted fried noodles was improved by applying interaction between pea protein isolate (PPI) and green tea extract (GTE). Radical-scavenging activities of GTE were stably maintained when exposed to acidic pH, UV light, and fluorescent light, but decreased by approximately 65% when exposed to 80°C for 168h. The RVA profiles of noodle dough showed that peak viscosity and breakdown increased significantly but that setback and final viscosity remained unchanged with 20% rice flour replacement. PPI significantly decreased the viscosity parameters of rice-supplemented dough, and the addition of GTE recovered these values significantly. The cooking loss and viscoelasticity (Rmax) of cooked rice-supplemented noodles were fully restored by combined treatment of PPI and GTE. GTE decreased the peroxide value of fried noodles by 14% after storage at 63°C for 16days. Therefore, PPI+GTE treatment has great potential for use in fried noodles owing to the reinforced network and antioxidant activity. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. A rapid method for isolation of purified, physiologically active chloroplasts, used to study the intracellular distribution of amino acids in pea leaves.

    PubMed

    Mills, W R; Joy, K W

    1980-02-01

    A procedure is described for the rapid (<5 min) isolation of purified, physiologically active chloroplasts from Pisum sativum L. Mitochondrial and microbody contamination is substantially reduced and broken chloroplasts are excluded by washing through a layer containing a treated silica sol. On average the preparations contain 93% intact chloroplasts and show high rates of (14)CO2 fixation and CO2-dependent O2 evolution (over 100 μmol/mg chlorophyll(chl)/h); they are also able to carry out light-driven incorporation of leucine into protein (4 nmol/mg chl/h). The amino-acid contents of chloroplasts prepared from leaves and from leaf protoplasts have been determined. Asparagine is the most abundant amino acid in the pea chloroplast (>240 nmol/mg chl), even thought it is proportionately lower in the chloroplast relative to the rest of the cell. The chloroplasts contain about 20% of many of the amino acids of the cell, but for individual amino acids the percentage in the chloroplast ranges from 8 to 40% of the cell total. Glutamic acid, glutamine and aspartic acid are enriched in the chloroplasts, while asparagine, homoserine and β-(isoxazolin-5-one-2-yl)-alanine are relatively lower. Leakage of amino acids from the chloroplast during preparation or repeated washing was ca. 20%. Some differences exist between the amino-acid composition of chloroplasts isolated from intact leaves and from protoplasts. In particular, γ-aminobutyric acid accumulates to high levels, while homoserine and glutamic acid decrease, during protoplast formation and breakage.

  3. Beta-ketoacyl-acyl carrier protein synthase III from pea (Pisum sativum L.): properties, inhibition by a novel thiolactomycin analogue and isolation of a cDNA clone encoding the enzyme.

    PubMed

    Jones, A Lesley; Gane, Andy M; Herbert, Derek; Willey, David L; Rutter, Andrew J; Kille, Peter; Dancer, Jane E; Harwood, John L

    2003-03-01

    A beta-ketoacyl-acyl carrier protein (ACP) synthase III (KAS III; short-chain condensing enzyme) has been partly purified from pea leaves. The enzyme, which had acetyl-CoA:ACP acyltransferase (ACAT) activity, was resolved from a second, specific, ACAT protein. The KAS III enzyme had a derived molecular mass of 42 kDa (from its cDNA sequence) and operated as a dimer. Its enzymological characteristics were similar to those of two other plant KAS III enzymes except for its inhibition by thiolactomycin. A derivative of thiolactomycin containing a longer (C8 saturated) hydrophobic side-chain (compound 332) was a more effective inhibitor of pea KAS III and showed competitive inhibition towards malonyl-ACP whereas thiolactomycin showed uncompetitive characteristics at high concentrations. This difference may be due to the better fit of compound 332 into a hydrophobic pocket at the active site. A full-length cDNA for the pea KAS III was isolated. This was expressed in Escherichia coli as a fusion protein with glutathione S-transferase in order to facilitate subsequent purification. Demonstrated activity in preparations from E. coli confirmed that the cDNA encoded a KAS III enzyme. Furthermore, the expressed KAS III had ACAT activity, showing that the latter was inherent. The derived amino acid sequence of the pea cDNA showed 81-87% similarity to that for other plant dicotyledon KAS IIIs, somewhat less for Allium porrum (leek, 71%) and for Porphyra spp. (62%), Synechocystis spp. (65%) and various bacteria (42-65%). The pea KAS III exhibited four areas of homology, three of which were around the active-site Cys(123), His(323) and Asn(353). In addition, a stretch of 23 amino acids (residues 207-229 in the pea KAS III) was almost completely conserved in the plant KAS IIIs. Modelling this stretch showed they belonged to a peptide fragment that fitted over the active site and contained segments suggested to be involved in substrate binding and in conformational changes during

  4. Pea Xyloglucan and Cellulose

    PubMed Central

    Hayashi, Takahisa; Marsden, Margery P. F.; Delmer, Deborah P.

    1987-01-01

    Since xyloglucan is believed to bind to cellulose microfibrils in the primary cell walls of higher plants and, when isolated from the walls, can also bind to cellulose in vitro, the binding mechanism of xyloglucan to cellulose was further investigated using radioiodinated pea xyloglucan. A time course for the binding showed that the radioiodinated xyloglucan continued to be bound for at least 4 hours at 40°C. Binding was inhibited above pH 6. Binding capacity was shown to vary for celluloses of different origin and was directly related to the relative surface area of the microfibrils. The binding of xyloglucan to cellulose was very specific and was not affected by the presence of a 10-fold excess of (1→2)-β-glucan, (1→3)-β-glucan, (1→6)-β-glucan, (1→3, 1→4)-β-glucan, arabinogalactan, or pectin. When xyloglucan (0.1%) was added to a cellulose-forming culture of Acetobacter xylinum, cellulose ribbon structure was partially disrupted indicating an association of xyloglucan with cellulose at the time of synthesis. Such a result suggests that the small size of primary wall microfibrils in higher plants may well be due to the binding of xyloglucan to cellulose during synthesis which prevents fasciation of small fibrils into larger bundles. Fluorescent xyloglucan was used to stain pea cell wall ghosts prepared to contain only the native xyloglucan:cellulose network or only cellulose. Ghosts containing only cellulose showed strong fluorescence when prepared before or after elongation; as predicted, the presence of native xyloglucan in the ghosts repressed binding of added fluorescent xyloglucan. Such ghosts, prepared after elongation when the ratio of native xyloglucan:cellulose is substantially reduced, still showed only faint fluorescence, indicating that microfibrils continue to be coated with xyloglucan throughout the growth period. Images Fig. 5 Fig. 6 PMID:16665254

  5. Resistance to moist conditions of whey protein isolate and pea starch biodegradable films and low density polyethylene nondegradable films: a comparative study

    NASA Astrophysics Data System (ADS)

    Mehyar, G. F.; Bawab, A. Al

    2015-10-01

    Biodegradable packaging materials are degraded under the natural environmental conditions. Therefore using them could alleviate the problem of plastics accumulation in nature. For effective replacement of plastics, with biodegradable materials, biodegradable packages should keep their properties under the high relative humidity (RH) conditions. Therefore the objectives of the study were to develop biodegradable packaging material based on whey protein isolate (WPI) and pea starch (PS). To study their mechanical, oxygen barrier and solubility properties under different RHs compared with those of low density polyethylene (LDPE), the most used plastic in packaging. Films of WPI and PS were prepared separately and conditioned at different RH (30-90%) then their properties were studied. At low RHs (<50%), WPI films had 2-3 times lower elongation at break (E or stretchability) than PS and LDPE. Increasing RH to 90% significantly (P<0.01) increased the elongation of PS but not WPI and LDPE films. LDPE and WPI films kept significantly (P<0.01) higher tensile strength (TS) than PS films at high RH (90%). Oxygen permeability (OP) of all films was very low (<0.5 cm3 μm m-2 d-1 kPa-1) below 40% RH but increased for PS films and became significantly (P<0.01) different than that of LDPE and WPI at > 40% RH. Oxygen permeability of WPI and LDPE did not adversely affected by increasing RH to 65%. Furthermore, WPI and LDPE films had lower degree of hydration at 50% and 90% RH and total soluble matter than PS films. These results suggest that WPI could be successfully replacing LDPE in packaging of moist products.

  6. Growth factor-dependent initiation of DNA replication in nuclei isolated from an interleukin 3-dependent murine myeloid cell line.

    PubMed

    Munshi, N C; Gabig, T G

    1990-01-01

    To study the proliferative response of hematopoietic cells to growth factors at the molecular level, we developed a cell-free system for growth factor-dependent initiation of genomic DNA replication. Nuclei were isolated from the IL-3-dependent cell line NFS/N1-H7 after a 10-h period of IL-3 deprivation. Cytosolic and membrane-containing subcellular fractions were prepared from proliferating NFS/N1-H7 cells. Nuclei from the nonproliferating cells (+/- IL-3) showed essentially no incorporation of [3H]thymidine during a 16-h incubation with a mixture of unlabeled GTP, ATP, UTP, CTP, dGTP, dATP, dCTP, and [3H]dTTP. When the combination of IL-3, a cytosolic fraction, and a membrane-containing fraction from proliferating cells was added to nuclei from nonproliferating cells, a burst of [3H]thymidine incorporation into DNA began after a 12-h lag period, attained a maximal rate at 16 h, and reached a level of 860 pmol thymidine/10(6) nuclei at 24 h (corresponding to replication of approximately 56% total mouse genomic DNA). This DNA synthesis was inhibited approximately 90% by the specific DNA polymerase alpha inhibitor aphidicolin. Deletion of a single cellular component or IL-3 from the system resulted in a marked reduction of DNA replication (-membrane, 80 +/- 4%; -cytosol, 90% +/- 4%; -IL-3, 74 +/- 7% inhibition). This model requires a growth factor (IL-3), a sedimentable cell fraction containing its receptor and possibly additional membrane-associated components, and a cytosolic fraction. It appears to recapitulate the molecular events required for progression from early G1 to S phase of the cell cycle induced by IL-3 binding to its receptor.

  7. Growth factor-dependent initiation of DNA replication in nuclei isolated from an interleukin 3-dependent murine myeloid cell line.

    PubMed Central

    Munshi, N C; Gabig, T G

    1990-01-01

    To study the proliferative response of hematopoietic cells to growth factors at the molecular level, we developed a cell-free system for growth factor-dependent initiation of genomic DNA replication. Nuclei were isolated from the IL-3-dependent cell line NFS/N1-H7 after a 10-h period of IL-3 deprivation. Cytosolic and membrane-containing subcellular fractions were prepared from proliferating NFS/N1-H7 cells. Nuclei from the nonproliferating cells (+/- IL-3) showed essentially no incorporation of [3H]thymidine during a 16-h incubation with a mixture of unlabeled GTP, ATP, UTP, CTP, dGTP, dATP, dCTP, and [3H]dTTP. When the combination of IL-3, a cytosolic fraction, and a membrane-containing fraction from proliferating cells was added to nuclei from nonproliferating cells, a burst of [3H]thymidine incorporation into DNA began after a 12-h lag period, attained a maximal rate at 16 h, and reached a level of 860 pmol thymidine/10(6) nuclei at 24 h (corresponding to replication of approximately 56% total mouse genomic DNA). This DNA synthesis was inhibited approximately 90% by the specific DNA polymerase alpha inhibitor aphidicolin. Deletion of a single cellular component or IL-3 from the system resulted in a marked reduction of DNA replication (-membrane, 80 +/- 4%; -cytosol, 90% +/- 4%; -IL-3, 74 +/- 7% inhibition). This model requires a growth factor (IL-3), a sedimentable cell fraction containing its receptor and possibly additional membrane-associated components, and a cytosolic fraction. It appears to recapitulate the molecular events required for progression from early G1 to S phase of the cell cycle induced by IL-3 binding to its receptor. Images PMID:2104881

  8. Pea weevil damage and chemical characteristics of pea cultivars determining their resistance to Bruchus pisorum L.

    PubMed

    Nikolova, I

    2016-04-01

    Bruchus pisorum (L.) is one of the most intractable pest problems of cultivated pea in Europe. Development of resistant cultivars is very important to environmental protection and would solve this problem to a great extent. Therefore, the resistance of five spring pea cultivars was studied to B. pisorum: Glyans, Modus; Kamerton and Svit and Pleven 4 based on the weevil damage and chemical composition of seeds. The seeds were classified as three types: healthy seeds (type one), damaged seeds with parasitoid emergence holes (type two) and damaged seeds with bruchid emergence holes (type three). From visibly damaged pea seeds by pea weevil B. pisorum was isolated the parasitoid Triaspis thoracica Curtis (Hymenoptera, Braconidae). Modus, followed by Glyans was outlined as resistant cultivars against the pea weevil. They had the lowest total damaged seed degree, loss in weight of damaged seeds (type two and type three) and values of susceptibility coefficients. A strong negative relationship (r = -0.838) between the weight of type one seeds and the proportion of type three seeds was found. Cultivars with lower protein and phosphorus (P) content had a lower level of damage. The crude protein, crude fiber and P content in damaged seeds significantly or no significantly were increased as compared with the healthy seeds due to weevil damage. The P content had the highest significant influence on pea weevil infestation. Use of chemical markers for resistance to the creation of new pea cultivars can be effective method for defense and control against B. pisorum.

  9. Metabolic and chemical properties of basic proteins isolated from nuclei of rat liver and thymus gland

    PubMed Central

    Ord, Margery G.; Raaf, J. H.; Smit, J. A.; Stocken, L. A.

    1965-01-01

    1. The effects of alkylating agents and disulphides on the thiol-containing proteins of nuclei from rat thymus and liver were studied. Three protein fractions were examined: histones extracted with 50mm- and 250mm-hydrochloric acid and the residual protein. None of the reagents selectively reacted with any one of the protein fractions. 2. Amino acid uptake in vitro into the histones of nuclei from rat thymus was analysed by preparative electrophoresis of the proteins extracted with 50mm- and 250mm-hydrochloric acid. After 1hr. at 37° the greater incorporation was into the proteins extracted with 50mm-hydrochloric acid. 3. Preparative electrophoresis was used to study the relative thiol contents of the proteins of the 50mm-hydrochloric acid extract from thymus nuclei by labelling the histones in vitro with 14C-labelled N-ethylmaleimide. 4. The capacity of the proteins extracted from rat thymus with 50mm- and 250mm-hydrochloric acid, and of the components from these extracts separated by preparative electrophoresis, to combine with DNA and to depress DNA-dependent RNA synthesis was studied. The histones extracted with 50mm-hydrochloric acid were more lysine-rich than those extracted with 250mm-hydrochloric acid. Wide variations were found in the abilities of the separated components to depress RNA synthesis. PMID:14340080

  10. Improved technique for fluorescence in situ hybridisation analysis of isolated nuclei from archival, B5 or formalin fixed, paraffin wax embedded tissue.

    PubMed

    Schurter, M J; LeBrun, D P; Harrison, K J

    2002-04-01

    Fluorescence in situ hybridisation (FISH) is an effective method to detect chromosomal alterations in a variety of tissue types, including archived paraffin wax embedded specimens fixed in B5 or formalin. However, precipitating fixatives such as B5 have been known to produce unsatisfactory results in comparison with formalin when used for FISH. This study describes an effective nuclear isolation and FISH procedure for B5 and formalin fixed tissue, optimising the nuclear isolation step and nuclei pretreatments using tonsil and mantle cell lymphoma specimens. The protocol presented can be used to isolate nuclei and perform FISH on B5 or formalin fixed, paraffin wax embedded samples from a variety of tissue types.

  11. Mechanisms of DNA strand breakage and interstrand cross-linking by diaziridinylbenzoquinone (diaziquone) in isolated nuclei from human cells.

    PubMed

    Szmigiero, L; Kohn, K W

    1984-10-01

    AZQ had been found to produce DNA strand breaks and interstrand cross-links in intact cells; evidence had indicated that these two DNA lesions arise by different chemical mechanisms and vary independently in degree in different cell types. In the present work, the mechanisms of the production of DNA strand breaks and interstrand cross-links by AZQ were studied in isolated cell nuclei. This system avoided the problem of poor penetration of test substances into cells. The DNA lesions were measured by means of the alkaline elution technique. It was found that the production of DNA strand breaks by AZQ in isolated nuclei required the addition of a reducing agent such as NADPH and was almost completely prevented by superoxide dismutase. This indicates that the mechanism of DNA strand breakage involves transfer of an electron from a reduced form of AZQ to molecular oxygen. Unexpectedly, interstrand cross-linking also was enhanced greatly by previous reduction of AZQ by NADPH or NaBH4. However, this reaction was not inhibited by superoxide dismutase. General alkylating activity of AZQ also was stimulated by reduction; the pH-dependence of this reaction was determined. The mechanism of DNA interstrand cross-linking by AZQ was surmised to stem from alkylation reactions of the two aziridine groups. The findings suggest the possibility that AZQ or related compounds may function as bioreductive alkylating agents which might be selectively toxic to hypoxic tissues.

  12. Role of chloroplast photosystems II and I in apoptosis of pea guard cells.

    PubMed

    Samuilov, V D; Lagunova, E M; Gostimsky, S A; Timofeev, K N; Gusev, M V

    2003-08-01

    Received Revision received We investigated the CN--induced apoptosis of guard cells in epidermal peels isolated from pea (Pisum sativum L.) leaves. This process was considerably stimulated by illumination and suppressed by the herbicides DCMU (an inhibitor of the electron transfer between quinones Q(A) and Q(B) in PS II) and methyl viologen (an electron acceptor from PS I). These data favor the conclusion drawn by us earlier that chloroplasts are involved in the apoptosis of guard cells. Pea mutants with impaired PS I (Chl-5), PS II (Chl-I), and PS II + PS I (Xa-17) were tested. Their lesions were confirmed by the ESR spectra of Signal I (oxidized PS I reaction centers) and Signal II (oxidized tyrosine residue Y(D) in PS II). Destruction of nuclei (a symptom of apoptosis) and their consecutive disappearance in guard cells were brought about by CN- in all the three mutants and in the normal pea plants. These results indicate that the light-induced enhancement of apoptosis of guard cells and its removal by DCMU are associated with PS II function. The effect of methyl viologen preventing CN--induced apoptosis in wild-type plants was removed or considerably decreased upon the impairment of the PS II and/or PS I activity.

  13. Generation of hydroxyl radical in isolated pea root cell wall, and the role of cell wall-bound peroxidase, Mn-SOD and phenolics in their production.

    PubMed

    Kukavica, Biljana; Mojovic, Milos; Vuccinic, Zeljko; Maksimovic, Vuk; Takahama, Umeo; Jovanovic, Sonja Veljovic

    2009-02-01

    The hydroxyl radical produced in the apoplast has been demonstrated to facilitate cell wall loosening during cell elongation. Cell wall-bound peroxidases (PODs) have been implicated in hydroxyl radical formation. For this mechanism, the apoplast or cell walls should contain the electron donors for (i) H(2)O(2) formation from dioxygen; and (ii) the POD-catalyzed reduction of H(2)O(2) to the hydroxyl radical. The aim of the work was to identify the electron donors in these reactions. In this report, hydroxyl radical (.OH) generation in the cell wall isolated from pea roots was detected in the absence of any exogenous reductants, suggesting that the plant cell wall possesses the capacity to generate .OH in situ. Distinct POD and Mn-superoxide dismutase (Mn-SOD) isoforms different from other cellular isoforms were shown by native gel electropho-resis to be preferably bound to the cell walls. Electron paramagnetic resonance (EPR) spectroscopy of cell wall isolates containing the spin-trapping reagent, 5-diethoxyphosphoryl-5-methyl-1-pyrroline-N-oxide (DEPMPO), was used for detection of and differentiation between .OH and the superoxide radical (O(2)(-).). The data obtained using POD inhibitors confirmed that tightly bound cell wall PODs are involved in DEPMPO/OH adduct formation. A decrease in DEPMPO/OH adduct formation in the presence of H(2)O(2) scavengers demonstrated that this hydroxyl radical was derived from H(2)O(2). During the generation of .OH, the concentration of quinhydrone structures (as detected by EPR spectroscopy) increased, suggesting that the H(2)O(2) required for the formation of .OH in isolated cell walls is produced during the reduction of O(2) by hydroxycinnamic acids. Cell wall isolates in which the proteins have been denaturated (including the endogenous POD and SOD) did not produce .OH. Addition of exogenous H(2)O(2) again induced the production of .OH, and these were shown to originate from the Fenton reaction with tightly bound metal ions

  14. Sphingolipid metabolites selectively elicit increases in nuclear calcium concentration in cell suspension cultures and in isolated nuclei of tobacco.

    PubMed

    Xiong, Tou Cheu; Coursol, Sylvie; Grat, Sabine; Ranjeva, Raoul; Mazars, Christian

    2008-01-01

    Sphingolipids are known to interfere with calcium-based signalling pathways. Here we report that these compounds modulate nuclear calcium signalling in tobacco BY-2 cells. Nuclear protein kinase activity phosphorylated endogenous sphingoid long-chain bases (LCBs), suggesting that LCBs are actively metabolized in the nucleus of tobacco BY-2 cells. The Delta4-unsaturated LCB D-erythro-sphingosine and the saturated LCB D-ribo-phytosphingosine elicited increases in free calcium in the nucleus in a dose-dependent and structure-related manner. However, neither sphingosine-1-phosphate nor C2-ceramide was able to stimulate nuclear calcium changes. N-,N-Dimethyl-D-erythro-sphingosine, a structural analogue of D-erythro-sphingosine, was the most efficient LCB so far tested in eliciting nuclear calcium changes both in intact tobacco BY-2 cells and in isolated nuclei. TRP channel inhibitors prevent the effect of DMS, suggesting that LCBs may activate TRP-like channels located on the inner nuclear membrane Collectively, the obtained data show that nuclei respond to LCBs on their own independently of the cytosolic compartment.

  15. Transcriptional activation of Xenopus class III genes in chromatin isolated from sperm and somatic nuclei.

    PubMed Central

    Wolffe, A P

    1989-01-01

    Xenopus sperm chromatin lacks class III transcription complexes and somatic histone H1. Inactive class III genes in sperm chromatin are easily programmed with transcription complexes de novo and transcribed in Xenopus oocyte nuclear extract. In contrast, repressed class III genes in somatic chromatin are not transcribed in the oocyte nuclear extract. Class III genes that are initially inactive or repressed in both types of chromatin can be efficiently transcribed in a cell free preparation of Xenopus eggs. Chromatin mediated repression of class III genes in somatic nuclei is reversible in Xenopus egg extract, but not in the oocyte nuclear extract. Any inhibition of transcription attributed to chromatin assembly onto a gene, will therefore depend on the extract in which transcription is assayed. Images PMID:2915929

  16. Pathogenicity of entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana (Hypocreales: Clavicipitaceae) isolates to the adult pea leafminer (Diptera: Agromyzidae) and prospects of an autoinoculation device for infection in the field.

    PubMed

    Migiro, L N; Maniania, N K; Chabi-Olaye, A; Vandenberg, J

    2010-04-01

    Seventeen isolates of Metarhizium anisopliae (Metschnikoff) Sorokin and three isolates of Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Clavicipitaceae) were evaluated for their pathogenicity to the adult pea leafminer, Liriomyza huidobrensis (Blanchard) (Diptera: Agromyzidae), in the laboratory. Flies were contaminated with dry conidia through a velvet material wrapped around the inner side of a cylindrical plastic tube. All the isolates were pathogenic to the pea leafminer, causing mortality between 40 and 100% at 5 d after exposure. The lethal time for 50% mortality (LT(50)) ranged from 2.6 to 5.4 d, whereas the LT(90) values varied between 3.2 and 9.1 d depending on the isolate. An autoinoculation device was evaluated in cage field experiments using only one of the virulent isolates, M. anisopliae ICIPE 20. The device was loaded with 2-3 g of dry conidia. Mortality of up to 100% was observed in flies captured from fungus-treated cages held under laboratory conditions. The average number of spores picked up by a single fly visiting the device increased with days after inoculation. One day after the inoculation, adults picked up an average of 4.1 +/- 0.7 x 10(5) conidia and 39.6 +/- 4.0 x 10(5) conidia 5 d after inoculation. Depending on the sampling date, the LT(50) varied between 1.8 and 3.4 d. Results indicate that some isolates of B. bassiana and M. anisopliae are highly pathogenic to L. huidobrensis, suggesting a potential for their use in the control of this pest. They also suggest the possibility of L. huidobrensis suppression with fungi using an autoinoculation device.

  17. Chromatin motion and nucleosome dynamics in live cells and isolated nuclei observed using two-photon standing wave photobleaching

    NASA Astrophysics Data System (ADS)

    Davis, Sara K.; Bardeen, Christopher

    2003-03-01

    DNA in eukaryotic cells is wrapped around histone proteins to form chromatin fibers. Fluctuations in the local structure of these chromatin fibers are believed to control the transcriptional availability of DNA. Such fluctuations also result in the constrained diffusive motion of the chromatin in live cells observed using high resolution fluorescence microscopy. Using a two-photon standing wave fluorescence photobleaching method, we have measured the short-range (100 nm) diffusive motion of labeled DNA both in vivo and in vitro. The motion observed in living cells is absent from isolated nuclei. By varying the ionic strength of nuclear environment, we can modify the binding of the core histones to the DNA. We can also use chemical or photochemical reactions to irreversibly crosslink the histones to the DNA. Both experiments provide evidence that chromatin diffusion is controlled by the chemistry of the histone interactions, as opposed to thermal fluctuations or polymer entanglement.

  18. Ascochyta blight of peas

    USDA-ARS?s Scientific Manuscript database

    Identification, taxonomy & nomenclature, epidemiology, symptoms, host ranges, and management are described for three fungal pathogens which collectively and individually cause Ascochyta blight of field pea (Pisum sativum): Ascochyta pisi, Ascochyta pinodes, and Ascochyta pinodella. The first two are...

  19. Peas (Pisum sativum L.).

    PubMed

    Grant, Jan; Cooper, Pauline

    2006-01-01

    In this chapter we describe a robust method for transformation of peas that has been successfully used in our laboratory since 1992. Using immature pea seed collected from field- or greenhouse-grown plants, we have produced transgenic lines for over 30 genotypes including named pea cultivars and advanced breeding lines. This method uses immature cotyledons as the explant, and the transformation efficiency is in the range 0.2 to 13.5% of cotyledons producing at least one independently transformed line. Agrobacterium tumefaciens strains AGL1 and KYRT1 are the most successful in our procedure, and kanamycin, phosphinothricin, and hygromycin are reliable selectable markers. Potentially useful genes have been introduced for pest and disease resistance, altering quality traits, and investigating metabolic pathways and are being studied in transgenic pea lines.

  20. Concentrations of long-chain acyl-acyl carrier proteins during fatty acid synthesis by chloroplasts isolated from pea (Pisum sativum), safflower (Carthamus tinctoris), and amaranthus (Amaranthus lividus) leaves

    SciTech Connect

    Roughan, G.; Nishida, I. )

    1990-01-01

    Fatty acid synthesis from (1-14C)acetate by chloroplasts isolated from peas and amaranthus was linear for at least 15 min, whereas incorporation of the tracer into long-chain acyl-acyl carrier protein (ACP) did not increase after 2-3 min. When reactions were transferred to the dark after 3-5 min, long-chain acyl-ACPs lost about 90% of their radioactivity and total fatty acids retained all of theirs. Half-lives of the long-chain acyl-ACPs were estimated to be 10-15 s. Concentrations of palmitoyl-, stearoyl-, and oleoyl-ACP as indicated by equilibrium labeling during steady-state fatty acid synthesis, ranged from 0.6-1.1, 0.2-0.7, and 0.4-1.6 microM, respectively, for peas and from 1.6-1.9, 1.3-2.6, and 0.6-1.4 microM, respectively, for amaranthus. These values are based on a chloroplast volume of 47 microliters/mg chlorophyll and varied according to the mode of the incubation. A slow increase in activity of the fatty acid synthetase in safflower chloroplasts resulted in long-chain acyl-ACPs continuing to incorporate labeled acetate for 10 min. Upon re-illumination following a dark break, however, both fatty acid synthetase activity and acyl-ACP concentrations increased very rapidly. Palmitoyl-ACP was present at concentrations up to 2.5 microM in safflower chloroplasts, whereas those of stearoyl- and oleoyl-ACPs were in the lower ranges measured for peas. Acyl-ACPs were routinely separated from extracts of chloroplasts that had been synthesising long-chain fatty acids from labeled acetate by a minor modification of the method of Mancha et al. The results compared favorably with those obtained using alternative analytical methods such as adsorption to filter paper and partition chromatography on silicic acid columns.

  1. Monitoring UVR induced damage in single cells and isolated nuclei using SR-FTIR microspectroscopy and 3D confocal Raman imaging.

    PubMed

    Lipiec, Ewelina; Bambery, Keith R; Heraud, Philip; Kwiatek, Wojciech M; McNaughton, Don; Tobin, Mark J; Vogel, Christian; Wood, Bayden R

    2014-09-07

    SR-FTIR in combination with Principal Component Analysis (PCA) was applied to investigate macromolecular changes in a population of melanocytes and their extracted nuclei induced by environmentally relevant fluxes of UVR (Ultraviolet Radiation). Living cells and isolated cellular nuclei were investigated post-irradiation for three different irradiation dosages (130, 1505, 15,052 Jm(-2) UVR, weighted) after either 24 or 48 hours of incubation. DNA conformational changes were observed in cells exposed to an artificial UVR solar-simulator source as evidenced by a shift in the DNA asymmetric phosphodiester vibration from 1236 cm(-1) to 1242 cm(-1) in the case of the exposed cells and from 1225 cm(-1) to 1242 cm(-1) for irradiated nuclei. PCA Scores plots revealed distinct clustering of spectra from irradiated cells and nuclei from non-irradiated controls in response to the range of applied UVR radiation doses. 3D Raman confocal imaging in combination with k-means cluster analysis was applied to study the effect of the UVR radiation exposure on cellular nuclei. Chemical changes associated with apoptosis were detected and included intra-nuclear lipid deposition along with chromatin condensation. The results reported here demonstrate the utility of SR-FTIR and Raman spectroscopy to probe in situ DNA damage in cell nuclei resulting from UVR exposure. These results are in agreement with the increasing body of evidence that lipid accumulation is a characteristic of aggressive cancer cells, and are involved in the production of membranes for rapid cell proliferation.

  2. Formation and rejoining of deoxyribonucleic acid double-strand breaks induced in isolated cell nuclei by antineoplastic intercalating agents.

    PubMed

    Pommier, Y; Schwartz, R E; Kohn, K W; Zwelling, L A

    1984-07-03

    The biochemical characteristics of the formation and disappearance of intercalator-induced DNA double-strand breaks (DSB) were studied in nuclei from mouse leukemia L1210 cells by using filter elution methodology [Bradley, M. O., & Kohn, K.W. (1979) Nucleic Acids Res. 7, 793-804]. The three intercalators used were 4'-(9-acridinylamino)-methanesulfon-m-anisidide (m-AMSA), 5-iminodaunorubicin (5-ID), and ellipticine. These compounds differ in that they produced predominantly DNA single-strand breaks (SSB) (m-AMSA) or predominantly DNA double-strand breaks (ellipticine) or a mixture of both SSB and DSB (5-ID) in whole cells. In isolated nuclei, each intercalator produced DSB at a frequency comparable to that which is produced in whole cells. Moreover, these DNA breaks reversed within 30 min after drug removal. It thus appeared that neither ATP nor other nucleotides were necessary for intercalator-dependent DNA nicking-closing reactions. The formation of the intercalator-induced DSB was reduced at ice temperature. Break formation was also reduced in the absence of magnesium, at a pH above 6.4 and at NaCl concentrations above 200 mM. In the presence of ATP and ATP analogues, the intercalator-induced cleavage was enhanced. These results suggest that the intercalator-induced DSB are enzymatically mediated and that the enzymes involved in these reactions can catalyze DNA double-strand cleavage and rejoining in the absence of ATP, although the occupancy of an ATP binding site might convert the enzyme to a form more reactive to intercalators. Three inhibitors of DNA topoisomerase II--novobiocin, nalidixic acid, and norfloxacin--reduced the formation of DNA strand breaks.(ABSTRACT TRUNCATED AT 250 WORDS)

  3. Multi-wavelength properties and smbh's masses of the isolated galaxies with active nuclei in the Local Universe

    NASA Astrophysics Data System (ADS)

    Vavilova, Iryna; Vasylenko, Anatolij; Babyk, Iuri; Pulatova, Nadya

    2016-07-01

    We apply the specially-oriented Astro-Space databases obtained with ground-based telescopes and space observatories to study the multi-wavelength spectral and physical properties of galaxies with active nuclei (AGNs), namely of isolated AGNs that are poorly investigated especially in X-rays. Such a study allowed us 1) to separate the internal evolution mechanisms from the environment influence and consider them as two separate processes related to fueling nuclear activity, 2) to explore absorption features and the X-ray continuum radiation from accretion disks around SMBHs (e.g. to select accretion models). In the case of detecting the Fe K emission line, it was possible to analyze the physical conditions in the AGNs innermost parts in more details. Using the SDSS spectral Hβ-line data we were able to estimate the SMBH masses of several isolated AGNs in the Local Universe, which are systematically lower than the SMBH masses of AGNs located in a dense environment. We present also the results of analysis of the spectral data obtained by XMM-Newton, Swift, Chandra, and INTEGRAL space observatories for several isolated AGNs from 2MIG catalogue, for which the available X-ray data were accessed. Among these objects are CGCG 179-005, NGC 6300, NGC 1050, NGC 2989, WKK 3050, ESO 438-009, ESO 317-038 and others. We determined corresponding spectral models and values of their parameters (spectral index, intrinsic absorption etc.). X-ray spectra for bright galaxies, NGC 6300 and Circinus, were analyzed up to 250 keV and their characteristics of emission features were determined in 6-7 keV range.

  4. A Single, Plastic Population of Mycosphaerella pinodes Causes Ascochyta Blight on Winter and Spring Peas (Pisum sativum) in France

    PubMed Central

    Guibert, Michèle; Leclerc, Aurélie; Andrivon, Didier; Tivoli, Bernard

    2012-01-01

    Plant diseases are caused by pathogen populations continuously subjected to evolutionary forces (genetic flow, selection, and recombination). Ascochyta blight, caused by Mycosphaerella pinodes, is one of the most damaging necrotrophic pathogens of field peas worldwide. In France, both winter and spring peas are cultivated. Although these crops overlap by about 4 months (March to June), primary Ascochyta blight infections are not synchronous on the two crops. This suggests that the disease could be due to two different M. pinodes populations, specialized on either winter or spring pea. To test this hypothesis, 144 pathogen isolates were collected in the field during the winter and spring growing seasons in Rennes (western France), and all the isolates were genotyped using amplified fragment length polymorphism (AFLP) markers. Furthermore, the pathogenicities of 33 isolates randomly chosen within the collection were tested on four pea genotypes (2 winter and 2 spring types) grown under three climatic regimes, simulating winter, late winter, and spring conditions. M. pinodes isolates from winter and spring peas were genetically polymorphic but not differentiated according to the type of cultivars. Isolates from winter pea were more pathogenic than isolates from spring pea on hosts raised under winter conditions, while isolates from spring pea were more pathogenic than those from winter pea on plants raised under spring conditions. These results show that disease developed on winter and spring peas was initiated by a single population of M. pinodes whose pathogenicity is a plastic trait modulated by the physiological status of the host plant. PMID:23023742

  5. A single, plastic population of Mycosphaerella pinodes causes ascochyta blight on winter and spring peas (Pisum sativum) in France.

    PubMed

    Le May, Christophe; Guibert, Michèle; Leclerc, Aurélie; Andrivon, Didier; Tivoli, Bernard

    2012-12-01

    Plant diseases are caused by pathogen populations continuously subjected to evolutionary forces (genetic flow, selection, and recombination). Ascochyta blight, caused by Mycosphaerella pinodes, is one of the most damaging necrotrophic pathogens of field peas worldwide. In France, both winter and spring peas are cultivated. Although these crops overlap by about 4 months (March to June), primary Ascochyta blight infections are not synchronous on the two crops. This suggests that the disease could be due to two different M. pinodes populations, specialized on either winter or spring pea. To test this hypothesis, 144 pathogen isolates were collected in the field during the winter and spring growing seasons in Rennes (western France), and all the isolates were genotyped using amplified fragment length polymorphism (AFLP) markers. Furthermore, the pathogenicities of 33 isolates randomly chosen within the collection were tested on four pea genotypes (2 winter and 2 spring types) grown under three climatic regimes, simulating winter, late winter, and spring conditions. M. pinodes isolates from winter and spring peas were genetically polymorphic but not differentiated according to the type of cultivars. Isolates from winter pea were more pathogenic than isolates from spring pea on hosts raised under winter conditions, while isolates from spring pea were more pathogenic than those from winter pea on plants raised under spring conditions. These results show that disease developed on winter and spring peas was initiated by a single population of M. pinodes whose pathogenicity is a plastic trait modulated by the physiological status of the host plant.

  6. Wild peas vary in their cross-compatibility with cultivated pea (Pisum sativum subsp. sativum L.) depending on alleles of a nuclear-cytoplasmic incompatibility locus.

    PubMed

    Bogdanova, V S; Kosterin, O E; Yadrikhinskiy, A K

    2014-05-01

    Divergent wild and endemic peas differ in hybrid sterility in reciprocal crosses with cultivated pea depending on alleles of a nuclear 'speciation gene' involved in nuclear-cytoplasmic compatibility. In hybrids between cultivated and wild peas, nuclear-cytoplasmic conflict frequently occurs. One of the nuclear genes involved, Scs1, was earlier mapped on Linkage Group III. In reciprocal crosses of seven divergent pea accessions with cultivated P. sativum, some alleles of Scs1 manifested incompatibility with an alien cytoplasm as a decrease in pollen fertility to about 50 % in the heterozygotes and lack of some genotypic classes among F2 segregants. Earlier, we defined monophyletic evolutionary lineages A, B, C and D of pea according to allelic state of three markers, from nuclear, plastid and mitochondrial genomes. All tested representatives of wild peas from the lineages A and C exhibited incompatibility due to Scs1 deleterious effects in crosses with testerlines of P. sativum subsp. sativum (the common cultivated pea) at least in one direction. A wild pea from the lineage B and a cultivated pea from the lineage D were compatible with the testerline in both directions. The tested accession of cultivated P. abyssinicum (lineage A) was partially compatible in both directions. The Scs1 alleles of some pea accessions even originating from the same geographic area were remarkably different in their compatibility with cultivated Pisum sativum cytoplasm. Variability of a gene involved in reproductive isolation is of important evolutionary role and nominate Scs1 as a speciation gene.

  7. X-rays from Green Pea analogues

    NASA Astrophysics Data System (ADS)

    Brorby, M.; Kaaret, P.

    2017-09-01

    X-ray observations of two metal-deficient luminous compact galaxies (LCG; SHOC 486 and SDSS J084220.94+115000.2) with properties similar to the so-called Green Pea galaxies were obtained using the Chandra X-ray Observatory. Green Pea galaxies are relatively small, compact (a few kpc across) galaxies that get their green colour from strong [O iii] λ5007 Å emission, an indicator of intense, recent star formation. These two galaxies were predicted to have the highest observed count rates, using the X-ray luminosity-star formation rate (LX-SFR) relation for X-ray binaries, from a statistically complete sample drawn from optical criteria. We determine the X-ray luminosity relative to SFR and metallicity for these two galaxies. Neither exhibits any evidence of active galactic nuclei, and we suspect that the X-ray emission originates from unresolved populations of high-mass X-ray binaries. We discuss the LX-SFR-metallicity plane for star-forming galaxies and show that the two LCGs are consistent with the prediction of this relation. This is the first detection of Green Pea analogues in X-rays.

  8. Pea Xyloglucan and Cellulose : IV. Assembly of beta-Glucans by Pea Protoplasts.

    PubMed

    Hayashi, T; Polonenko, D R; Camirand, A; Maclachlan, G

    1986-09-01

    The synthesis and assembly of xyloglucan were examined during early stages of wall regeneration by protoplasts isolated from growing regions of etiolated peas. During early stages of cultivation, fluorescence microscopy showed that the protoplast surface bound Calcofluor and ammonium salt of 8-anilino-1-naphthalene sulfonic acid and, in time, it also bound fluorescent fucose-binding lectin. Based on chemical analysis, 1,3-beta-glucan was the main polysaccharide formed by protoplasts and xyloglucan and cellulose were minor wall components. Binding between cellulose and xyloglucan was not as strong as that in tissues of intact pea plants, i.e. mild alkali could dissolve most xyloglucan from the protoplast. However, the addition of exogenous pea xyloglucan into the culture medium stimulated the deposition of new polysaccharides into the protoplast wall and enhanced the close association of newly formed xyloglucan with cellulose.

  9. Linkage Maps in Pea

    PubMed Central

    Ellis, THN.; Turner, L.; Hellens, R. P.; Lee, D.; Harker, C. L.; Enard, C.; Domoney, C.; Davies, D. R.

    1992-01-01

    We have analyzed segregation patterns of markers among the late generation progeny of several crosses of pea. From the patterns of association of these markers we have deduced linkage orders. Salient features of these linkages are discussed, as is the relationship between the data presented here and previously published genetic and cytogenetic data. PMID:1551583

  10. Pea (Pisum sativum L.)

    USDA-ARS?s Scientific Manuscript database

    Pea belongs to the Leguminosae plant family, the third largest flowering plant family with 800 genera and over 18,000 species. Tribe Fabeae is considered one of the youngest groups in the legumes and Bayesian molecular clock and ancestral range analysis suggest a crown age of 23 – 16 Mya, in the mi...

  11. Spermine stimulation of a nuclear NII kinase from pea plumules and its role in the phosphorylation of a nuclear polypeptide

    NASA Technical Reports Server (NTRS)

    Datta, N.; Schell, M. B.; Roux, S. J.

    1987-01-01

    We have previously demonstrated that spermine stimulates the phosphorylation of a 47 kilodalton nuclear polypeptide from pea plumules (N Datta, LK Hardison, SJ Roux 1986 Plant Physiol 82: 681-684). In this paper we report that spermine stimulates the activity of a cyclic AMP independent casein kinase, partially purified from a chromatin fraction of pea plumule nuclei. This effect of spermine was substrate specific; i.e. with casein as substrate, spermine stimulated the kinase activity, and with phosvitin as substrate, spermine completely inhibited the activity. The stimulation by spermine of the casein kinase was, in part, due to the lowering of the Mg2+ requirement of the kinase. Heparin could partially inhibit this casein kinase activity and spermine completely overcame this inhibition. By further purification of the casein kinase extract on high performance liquid chromatography, we fractionated it into an NI and an NII kinase. Spermine stimulated the NII kinase by 5- to 6-fold but had no effect on the NI kinase. Using [gamma-32P]GTP, we have shown that spermine promotes the phosphorylation of the 47 kilodalton polypeptide(s) in isolated nuclei, at least in part by stimulating an NII kinase.

  12. Spermine stimulation of a nuclear NII kinase from pea plumules and its role in the phosphorylation of a nuclear polypeptide

    NASA Technical Reports Server (NTRS)

    Datta, N.; Schell, M. B.; Roux, S. J.

    1987-01-01

    We have previously demonstrated that spermine stimulates the phosphorylation of a 47 kilodalton nuclear polypeptide from pea plumules (N Datta, LK Hardison, SJ Roux 1986 Plant Physiol 82: 681-684). In this paper we report that spermine stimulates the activity of a cyclic AMP independent casein kinase, partially purified from a chromatin fraction of pea plumule nuclei. This effect of spermine was substrate specific; i.e. with casein as substrate, spermine stimulated the kinase activity, and with phosvitin as substrate, spermine completely inhibited the activity. The stimulation by spermine of the casein kinase was, in part, due to the lowering of the Mg2+ requirement of the kinase. Heparin could partially inhibit this casein kinase activity and spermine completely overcame this inhibition. By further purification of the casein kinase extract on high performance liquid chromatography, we fractionated it into an NI and an NII kinase. Spermine stimulated the NII kinase by 5- to 6-fold but had no effect on the NI kinase. Using [gamma-32P]GTP, we have shown that spermine promotes the phosphorylation of the 47 kilodalton polypeptide(s) in isolated nuclei, at least in part by stimulating an NII kinase.

  13. Attraction of pea moth Cydia nigricana to pea flower volatiles.

    PubMed

    Thöming, Gunda; Knudsen, Geir K

    2014-04-01

    The pea moth Cydia nigricana causes major crop losses in pea (Pisum sativum) production. We investigated attraction of C. nigricana females to synthetic pea flower volatiles in a wind tunnel and in the field. We performed electroantennogram analysis on 27 previously identified pea plant volatiles, which confirmed antennal responses to nine of the compounds identified in pea flowers. A dose-dependent response was found to eight of the compounds. Various blends of the nine pea flower volatiles eliciting antennal responses were subsequently studied in a wind tunnel. A four-compound blend comprising hexan-1-ol, (E)-2-hexen-1-ol, (Z)-β-ocimene and (E)-β-ocimene was equally attractive to mated C. nigricana females as the full pea flower mimic blend. We conducted wind-tunnel tests on different blends of these four pea flower compounds mixed with a headspace sample of non-flowering pea plants. By considering the effects of such green leaf background odour, we were able to identify (Z)- and (E)-β-ocimene as fundamental for host location by the pea moths, and hexan-1-ol and (E)-2-hexen-1-ol as being of secondary importance in that context. In the field, the two isomers of β-ocimene resulted in trap catches similar to those obtained with the full pea flower mimic and the four-compound blend, which clearly demonstrated the prime significance of the β-ocimenes as attractants of C. nigricana. The high level of the trap catches of female C. nigricana noted in this first field experiment gives a first indication of the potential use of such artificial kairomones in pea moth control. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Immunological and biochemical evidence for nuclear localization of annexin in peas

    NASA Technical Reports Server (NTRS)

    Clark, G. B.; Dauwalder, M.; Roux, S. J.

    1998-01-01

    Immunofluorescent localization of annexins using an anti-pea annexin polyclonal antibody (anti-p35) in pea (Pisum sativum) leaf and stem epidermal peels showed staining of the nuclei and the cell periphery. Nuclear staining was also seen in cell teases prepared from pea plumules. The amount of nuclear stain was reduced both by fixation time and by dehydration and organic solvent treatment. Observation with confocal microscopy demonstrated that the anti-p35 stain was diffusely distributed throughout the nuclear structure. Immunoblots of purified nuclei, nuclear envelope matrix, nucleolar, and chromatin fractions showed a cross-reactive protein band of 35 kDa. These data are the first to show annexins localized in plant cell nuclei where they may play a role in nuclear function.

  15. Immunological and biochemical evidence for nuclear localization of annexin in peas

    NASA Technical Reports Server (NTRS)

    Clark, G. B.; Dauwalder, M.; Roux, S. J.

    1998-01-01

    Immunofluorescent localization of annexins using an anti-pea annexin polyclonal antibody (anti-p35) in pea (Pisum sativum) leaf and stem epidermal peels showed staining of the nuclei and the cell periphery. Nuclear staining was also seen in cell teases prepared from pea plumules. The amount of nuclear stain was reduced both by fixation time and by dehydration and organic solvent treatment. Observation with confocal microscopy demonstrated that the anti-p35 stain was diffusely distributed throughout the nuclear structure. Immunoblots of purified nuclei, nuclear envelope matrix, nucleolar, and chromatin fractions showed a cross-reactive protein band of 35 kDa. These data are the first to show annexins localized in plant cell nuclei where they may play a role in nuclear function.

  16. Isolation of Catharanthus roseus (L.) G. Don Nuclei and Measurement of Rate of Tryptophan decarboxylase Gene Transcription Using Nuclear Run-On Transcription Assay.

    PubMed

    Kumar, Santosh; Bhatia, Sabhyata

    2015-01-01

    An accurate assessment of transcription 'rate' is often desired to describe the promoter activity. In plants, isolation of transcriptionally active nuclei and their subsequent use in nuclear run-on assays has been challenging and therefore limit an accurate measurement of gene transcription 'rate'. Catharanthus roseus has emerged as a model medicinal plant as it exhibits an unsurpassed spectrum of chemodiversity, producing over 130 alkaloids through the terpenoid indole alkaloid (TIA) pathway and therefore serves as a 'molecular hub' to understand gene expression profiles. The protocols presented here streamline, adapt and optimize the existing methods of nuclear run-on assay for use in C. roseus. Here, we fully describe all the steps to isolate transcriptionally active nuclei from C. roseus leaves and utilize them to perform nuclear run-on transcription assay. Nuclei isolated by this method transcribed at a level consistent with their response to external stimuli, as transcription rate of TDC gene was found to be higher in response to external stimuli i.e. when seedlings were subjected to UV-B light or to methyl jasmonate (MeJA). However, the relative transcript abundance measured parallel through qRT-PCR was found to be inconsistent with the synthesis rate indicating that some post transcriptional events might have a role in transcript stability in response to stimuli. Our study provides an optimized, efficient and inexpensive method of isolation of intact nuclei and nuclear 'run-on' transcription assay to carry out in-situ measurement of gene transcription rate in Catharanthus roseus. This would be valuable in investigating the transcriptional and post transcriptional response of other TIA pathway genes in C. roseus. Isolated nuclei may also provide a resource that could be used for performing the chip assay as well as serve as the source of nuclear proteins for in-vitro EMSA studies. Moreover, nascent nuclear run-on transcript could be further subjected to RNA

  17. THE ORIGIN AND OPTICAL DEPTH OF IONIZING RADIATION IN THE 'GREEN PEA' GALAXIES

    SciTech Connect

    Jaskot, A. E.; Oey, M. S.

    2013-04-01

    Although Lyman-continuum (LyC) radiation from star-forming galaxies likely drove the reionization of the universe, observations of star-forming galaxies at low redshift generally indicate low LyC escape fractions. However, the extreme [O III]/[O II] ratios of the z = 0.1-0.3 Green Pea galaxies may be due to high escape fractions of ionizing radiation. To analyze the LyC optical depths and ionizing sources of these rare, compact starbursts, we compare nebular photoionization and stellar population models with observed emission lines in the Peas' Sloan Digital Sky Survey (SDSS) spectra. We focus on the six most extreme Green Peas, the galaxies with the highest [O III]/[O II] ratios and the best candidates for escaping ionizing radiation. The Balmer line equivalent widths and He I {lambda}3819 emission in the extreme Peas support young ages of 3-5 Myr, and He II {lambda}4686 emission in five extreme Peas signals the presence of hard ionizing sources. Ionization by active galactic nuclei or high-mass X-ray binaries is inconsistent with the Peas' line ratios and ages. Although stacked spectra reveal no Wolf-Rayet (WR) features, we tentatively detect WR features in the SDSS spectra of three extreme Peas. Based on the Peas' ages and line ratios, we find that WR stars, chemically homogeneous O stars, or shocks could produce the observed He II emission. If hot stars are responsible, then the Peas' optical depths are ambiguous. However, accounting for emission from shocks lowers the inferred optical depth and suggests that the Peas may be optically thin. The Peas' ages likely optimize the escape of LyC radiation; they are old enough for supernovae and stellar winds to reshape the interstellar medium, but young enough to possess large numbers of UV-luminous O or WR stars.

  18. Influence of steroid-hormone-receptor-protein complexes on initiation of ribonucleic acid synthesis in liver nuclei isolated from rats of various ages.

    PubMed Central

    Miller, J K; Bolla, R

    1981-01-01

    The effect of age on the induction of the initiation of RNA synthesis was investigated in liver nuclei isolated from adrenalectomized rats of various ages after binding of a dexamethasone-receptor-protein complex. Binding of this complex to nuclear chromatin resulted in increased initiation of nuclear RNA synthesis at all ages; however, an age-associated decline in the extent of this induction was observed. This suggests an age-related decrease of total rat liver nuclear RNA synthesis with a decreased response to glucocorticoid hormones. PMID:6171268

  19. Pea Chaperones under Centrifugation

    NASA Astrophysics Data System (ADS)

    Talalaiev, Oleksandr

    2008-06-01

    Etiolated Pisum sativum seedlings were subjected to altered g-forces by centrifugation (3-14g). By using semiquantitative RT-PCR, we studied transcripts of pea genes coding for chaperones that are representatives of small heat shock proteins (sHsps) family. Four members from the different classes of sHsps: cytosolic Hsp17.7 and Hsp18.1 (class I and class II accordingly), chloroplast Hsp21 (class III) and endoplasmic reticulum Hsp22.7 (class IV) were investigated. We conclude that exposure to 3, 7, 10 and 14g for 1h did not affect the level of sHsp transcripts.

  20. Isolation of Plant Nuclei at Defined Cell Cycle Stages Using EdU Labeling and Flow Cytometry.

    PubMed

    Wear, Emily E; Concia, Lorenzo; Brooks, Ashley M; Markham, Emily A; Lee, Tae-Jin; Allen, George C; Thompson, William F; Hanley-Bowdoin, Linda

    2016-01-01

    5-Ethynyl-2'-deoxyuridine (EdU) is a nucleoside analog of thymidine that can be rapidly incorporated into replicating DNA in vivo and, subsequently, detected by using "click" chemistry to couple its terminal alkyne group to fluorescent azides such as Alexa Fluor 488. Recently, EdU incorporation followed by coupling with a fluorophore has been used to visualize newly synthesized DNA in a wide range of plant species. One particularly useful application is in flow cytometry, where two-parameter sorting can be employed to analyze different phases of the cell cycle, as defined both by total DNA content and the amount of EdU pulse-labeled DNA. This approach allows analysis of the cell cycle without the need for synchronous cell populations, which can be difficult to obtain in many plant systems. The approach presented here, which was developed for fixed, EdU-labeled nuclei, can be used to prepare analytical profiles as well as to make highly purified preparations of G1, S, or G2/M phase nuclei for molecular or biochemical analysis. We present protocols for EdU pulse labeling, tissue fixation and harvesting, nuclei preparation, and flow sorting. Although developed for Arabidopsis suspension cells and maize root tips, these protocols should be modifiable to many other plant systems.

  1. New anthocyanins from purple pods of pea (Pisum spp.).

    PubMed

    Terahara, N; Honda, T; Hayashi, M; Ishimaru, K

    2000-12-01

    Two new anthocyanins were isolated from purple pods of pea (Pisum spp.). Their structures were identified as delphinidin 3-xylosylgalactoside-5-acetylglucoside and its deacetylated derivative by the usual chemical degradation methods and by spectroscopic methods such as UV-VIS, MS and NMR. Both pigments showed moderate stability and antioxidative activity in a neutral aqueous solution.

  2. Diversity of Rhizobium leguminosarum from pea fields in Washington State

    USDA-ARS?s Scientific Manuscript database

    Rhizobia-mediated biological nitrogen (N) fixation in legumes contributes to yield potential in these crops and also provides residual fertilizer to subsequent cereals. Our objectives were to collect isolates of Rhizobium leguminosarum from several pea fields in Washington, examine genetic diversity...

  3. A diagnostic guide for Fusarium Root Rot of pea

    USDA-ARS?s Scientific Manuscript database

    Fusarium root rot, caused by Fusarium solani f. sp. pisi, is a major root rot pathogen in pea production areas worldwide. Here we provide a diagnostic guide that describes: the taxonomy of the pathogen, signs and symptoms of the pathogen, host range, geographic distribution, methods used to isolate ...

  4. Physicochemical properties and in vitro digestibility of flour and starch from pea (Pisum sativum L.) cultivars.

    PubMed

    Chung, Hyun-Jung; Liu, Qiang

    2012-01-01

    Flours and isolated starches from three different cultivars (1544-8, 1658-11 and 1760-8) of pea grown under identical environmental conditions were evaluated for their physicochemical properties and in vitro digestibility. The protein content, total starch content and apparent amylose content of pea flour ranged from 24.4 to 26.3%, 48.8 to 50.2%, and 13.9 to 16.7%, respectively. In pea starches, the 1760-8 showed higher apparent amylose content and total starch content than the other cultivars. Pea starch granules were irregularly shaped, ranging from oval to round with a smooth surface. All pea starches showed C-type X-ray diffraction pattern with relative crystallinity ranging between 23.7 and 24.7%. Pea starch had only a single endothermic transition (12.1-14.2 J/g) in the DSC thermogram, whereas pea flour showed two separate endothermic transitions corresponding to starch gelatinization (4.54-4.71 J/g) and disruption of the amylose-lipid complex (0.36-0.78 J/g). In pea cultivars, the 1760-8 had significantly higher setback and final viscosity than the other cultivars in both pea flour (672 and 1170cP, respectively) and isolated starch (2901 and 4811cP). The average branch chain length of pea starches ranged from 20.1 to 20.3. The 1760-8 displayed a larger proportion of short branch chains, DP (degree of polymerization) 6-12 (21.1%), and a smaller proportion of long branch chains, DP≥37 (8.4%). The RDS, SDS and RS contents of pea flour ranged from 23.7 to 24.1%, 11.3 to 12.8%, and 13.2 to 14.8%, respectively. In pea starches, the 1760-8 showed a lower RDS content but higher SDS and RS contents. The expected glycemic index (eGI), based on the hydrolysis index, ranged from 36.9 to 37.7 and 69.8 to 70.7 for pea flour and isolated pea starch, respectively.

  5. Rapid activation of protein kinase C in isolated rat liver nuclei by prolactin, a known hepatic mitogen.

    PubMed Central

    Buckley, A R; Crowe, P D; Russell, D H

    1988-01-01

    Rat liver nuclei pure by enzymatic and electron microscope criteria contain protein kinase C (PKC) that can be activated several hundredfold within 3 min of addition of prolactin or phorbol 12-tetradecanoate 13-acetate. Rat prolactin stimulated PKC maximally at 10(-12) M, whereas ovine prolactin was maximally stimulatory at 10(-10) M. Activation was time and dose dependent, exhibited a biphasic pattern, and was blocked by anti-prolactin antiserum, by PKC inhibitors such as 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7) and sphingosine, and by cyclosporine. Moreover, the ability of prolactin to activate nuclear PKC was inhibited totally by a monoclonal antibody to the rat liver prolactin receptor, implicating a prolactin receptor-mediated activation process. Epidermal growth factor (EGF), a liver mitogen, caused a lesser but significant activation of nuclear PKC. However, EGF and suboptimal prolactin were synergistic. Human growth hormone, which has lactogenic properties, stimulated PKC activity, whereas nonlactogenic substances such as ovine growth hormone, insulin, dexamethasone, and 8-bromo-cAMP were inactive. That this may be a general mechanism for prolactin is suggested by the ability of prolactin to stimulate PKC 140-fold in rat splenocyte nuclei. Prolactin has comitogenic properties in lymphocytes. Images PMID:3186750

  6. The apparent absence of a pathway for synthesis of acetyl coenzyme A in pea chloroplasts: Lack of (14)CO 2 incorporation into lipids by the isolated organelle.

    PubMed

    Sherratt, D; Givan, C V

    1973-03-01

    We have examined the extent to which isotopic lable derived from photosynthetically fixed (14)CO2 can be transferred to lipids by aqueously isolated chloroplasts of Pisum sativum. Although photosynthetically active, chloroplast preparations incubated with (14)CO2 showed little or no accumulation of label in lipids under any condition tested. Under identical conditions the chloroplasts were readily able to incorporate [(14)C]acetate into the lipid fraction; a fatty-acid synthesizing system was therefore operative in these chloroplasts.The essential failure of the isolated chloroplasts to incorporate label from fixed (14)CO2 into fatty acids supports the view that the organelle itself does not possess a self-contained pathway for the synthesis of acetyl coenzyme A, and favours the possibility that a shuttle mechanism involving the participation of extra-chloroplastic enzymes may be responsible for supplying the chloroplast with acetyl coenzyme A in vivo.

  7. Draft Genome Sequence of the Plant Growth-Promoting Rhizobacterium Pseudomonas fluorescens Strain CREA-C16 Isolated from Pea (Pisum sativum L.) Rhizosphere

    PubMed Central

    Sorrentino, Roberto; Scotti, Riccardo; Salzano, Melania; Aurilia, Vincenzo

    2017-01-01

    ABSTRACT Herein, we report the draft genome sequence of Pseudomonas fluorescens strain CREA-C16, a plant growth-promoting rhizobacterium that was isolated from the rhizosphere of Pisum sativum L. plants. The genome sequence is ~6 Mb in size, with a G+C content of 60.1%, and includes 4,457 candidate protein-encoding genes. PMID:28126933

  8. Draft Genome Sequence of the Plant Growth-Promoting Rhizobacterium Pseudomonas fluorescens Strain CREA-C16 Isolated from Pea (Pisum sativum L.) Rhizosphere.

    PubMed

    D'Agostino, Nunzio; Sorrentino, Roberto; Scotti, Riccardo; Salzano, Melania; Aurilia, Vincenzo; Zaccardelli, Massimo

    2017-01-26

    Herein, we report the draft genome sequence of Pseudomonas fluorescens strain CREA-C16, a plant growth-promoting rhizobacterium that was isolated from the rhizosphere of Pisum sativum L. plants. The genome sequence is ~6 Mb in size, with a G+C content of 60.1%, and includes 4,457 candidate protein-encoding genes. Copyright © 2017 D’Agostino et al.

  9. The plastid ndh genes code for an NADH-specific dehydrogenase: isolation of a complex I analogue from pea thylakoid membranes.

    PubMed

    Sazanov, L A; Burrows, P A; Nixon, P J

    1998-02-03

    The plastid genomes of several plants contain ndh genes-homologues of genes encoding subunits of the proton-pumping NADH:ubiquinone oxidoreductase, or complex I, involved in respiration in mitochondria and eubacteria. From sequence similarities with these genes, the ndh gene products have been suggested to form a large protein complex (Ndh complex); however, the structure and function of this complex remains to be established. Herein we report the isolation of the Ndh complex from the chloroplasts of the higher plant Pisum sativum. The purification procedure involved selective solubilization of the thylakoid membrane with dodecyl maltoside, followed by two anion-exchange chromatography steps and one size-exclusion chromatography step. The isolated Ndh complex has an apparent total molecular mass of approximately 550 kDa and according to SDS/PAGE consists of at least 16 subunits including NdhA, NdhI, NdhJ, NdhK, and NdhH, which were identified by N-terminal sequencing and immunoblotting. The Ndh complex showed an NADH- and deamino-NADH-specific dehydrogenase activity, characteristic of complex I, when either ferricyanide or the quinones menadione and duroquinone were used as electron acceptors. This study describes the isolation of the chloroplast analogue of the respiratory complex I and provides direct evidence for the function of the plastid Ndh complex as an NADH:plastoquinone oxidoreductase. Our results are compatible with a dual role for the Ndh complex in the chlororespiratory and cyclic photophosphorylation pathways.

  10. Two parametric cell cycle analyses of plant cell suspension cultures with fragile, isolated nuclei to investigate heterogeneity in growth of batch cultivations.

    PubMed

    Haas, Christiane; Hegner, Richard; Helbig, Karsten; Bartels, Kristin; Bley, Thomas; Weber, Jost

    2016-06-01

    Plant cell suspensions are frequently considered to be heterogeneous with respect to growth in terms of progression of the cells through the cell cycle and biomass accumulation. Thus, segregated data of fractions in different cycle phases during cultivation is needed to develop robust production processes. Bromodeoxyuridine (BrdU) incorporation and BrdU-antibodies or 5-ethynyl-2'-deoxyuridine (EdU) click-it chemistry are frequently used to acquire such information. However, their use requires centrifugation steps that cannot be readily applied to sensitive cells, particularly if nuclei have to be extracted from the protective cellular milieu and envelopes for DNA analysis. Therefore, we have established a BrdU-Hoechst stain quenching protocol for analyzing nuclei directly isolated from delicate plant cell suspension cultures. After adding BrdU to test Harpagophytum procumbens cell suspension cultures the cell cycle distribution could be adequately resolved using its incorporation for the following 72 h (after which BrdU slowed biomass accumulation). Despite this limitation, the protocol allows resolution of the cell cycle distribution of cultures that cannot be analyzed using commonly applied methods due to the cells' fragility. The presented protocol enabled analysis of cycling heterogeneities in H. procumbens batch cultivations, and thus should facilitate process control of secondary metabolite production from fragile plant in vitro cultures. Biotechnol. Bioeng. 2016;113: 1244-1250. © 2015 Wiley Periodicals, Inc.

  11. 7 CFR 457.140 - Dry pea crop insurance provisions.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... crop. Dry peas. Peas (Pisum sativum L.), Austrian Peas (Pisum sativum spp arvense), Lentils (Lens... with grading under the United States Standards for Whole Dry Peas, Split Peas and Lentils will not be... example, if damaged smooth green and yellow pea acreage is replanted to lentils, the price election and...

  12. 7 CFR 457.140 - Dry pea crop insurance provisions.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... crop. Dry peas. Peas (Pisum sativum L.), Austrian Peas (Pisum sativum spp arvense), Lentils (Lens... with grading under the United States Standards for Whole Dry Peas, Split Peas and Lentils will not be... example, if damaged smooth green and yellow pea acreage is replanted to lentils, the price election and...

  13. 78 FR 68410 - United States Standards for Whole Dry Peas

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-14

    ... Lentil Handbook, which is available on GIPSA's public Web site at: http://www.gipsa.usda.gov/Publications... under the AMA include those for rice, whole dry peas, split peas, feed peas, lentils and beans. The U.S. standards for whole dry peas, split peas, feed peas, lentils and beans no longer appear in the Code of...

  14. 7 CFR 457.140 - Dry pea crop insurance provisions.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... crop. Dry peas. Peas (Pisum sativum L.), Austrian Peas (Pisum sativum spp arvense), Lentils (Lens... with grading under the United States Standards for Whole Dry Peas, Split Peas and Lentils will not be... example, if damaged smooth green and yellow pea acreage is replanted to lentils, the price election and...

  15. Protocol: Optimised methodology for isolation of nuclei from leaves of species in the Solanaceae and Rosaceae families

    PubMed Central

    2013-01-01

    In this study, a protocol is described for rapid preparation of an enriched, reasonably pure fraction of nuclear proteins from the leaves of tobacco (Nicotiana tabacum), potato (Solanum tuberosum) and apple (Malus domestica). The protocol gives reproducible results and can be carried out quickly in 2 hours. Tissue extracts clarified with filtration were treated with non-ionic detergent (Triton X-100) to lyse membranes of contaminating organelles. Nuclei were collected from a 60% Percoll layer of density gradient following low-speed centrifugation. Western blot analysis using antibodies to marker proteins of organelles indicated that the nuclear protein fractions were highly enriched and free or nearly free of proteins from the endoplasmic reticulum and chloroplasts. PMID:23886449

  16. Strains of Pseudomonas syringae pv. syringae from pea are phylogenetically and pathogenically diverse.

    PubMed

    Martín-Sanz, Alberto; de la Vega, Marcelino Pérez; Murillo, Jesús; Caminero, Constantino

    2013-07-01

    Pseudomonas syringae pv. syringae causes extensive yield losses in the pea crop worldwide, although there is little information on its host specialization and its interactions with pea. A collection of 88 putative P. syringae pv. syringae strains (including 39 strains isolated from pea) was characterized by repetitive polymerase chain reaction (rep-PCR), multilocus sequence typing (MLST), and syrB amplification and evaluated for pathogenicity and virulence. rep-PCR data grouped the strains from pea into two groups (1B and 1C) together with strains from other hosts; a third group (1A) was formed exclusively with strains isolated from non-legume species. MLST data included all strains from pea in the genomospecies 1 of P. syringae pathovars defined in previous studies; they were distributed in the same three groups defined by rep-PCR. The inoculations performed in two pea cultivars showed that P. syringae pv. syringae strains from groups 1A and 1C were less virulent than strains from group 1B, suggesting a possible pathogenic specialization in this group. This study shows the existence of genetically and pathogenically distinct P. syringae pv. syringae strain groups from pea, which will be useful for the diagnostic and epidemiology of this pathogen and for disease resistance breeding.

  17. 78 FR 63160 - United States Standards for Feed Peas, Split Peas, and Lentils

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-10-23

    ... Peas, and Lentils AGENCY: Grain Inspection, Packers and Stockyards Administration, USDA ACTION: Notice... Peas, and Lentils under the Agriculture Marketing Act (AMA) of 1946. To ensure that the standards and... current U.S. Standards for Feed Peas, Split Peas, and Lentils are meeting the needs in today's marketing...

  18. Kinetic studies of pea carbonic anhydrase.

    PubMed

    Johansson, I M; Forsman, C

    1993-12-01

    Chloroplast carbonic anhydrase from Pisum sativum has been isolated. The kinetic properties of the enzyme have been studied and comparisons to the well characterised human carbonic anhydrase II made. Pea carbonic anhydrase was found to be dependent on a reducing agent in order to retain the catalytic activity. Oxidised, inactive, enzyme could be activated by the addition of a SH-agent. However, such activation gave only 60% of the activity of an enzyme kept in a reduced state all the time. The kinetics of CO2 hydration show an increase in kcat as well as in kcat/Km with pH, but the pH profile does not follow a simple titration curve. The pH dependence is more complicated and it seems as if there are several titratable groups affecting the activity. At pH 9 we obtain a turnover number of 4 x 10(5) s-1 and a kcat/Km value of 1.8 x 10(8) M-1 s-1 with reference to the subunit. We also find that the enzyme needs high concentrations of buffer to work at a maximal rate. Apparent Km values with respect to the total buffer concentration are found between 52-185 mM at neutral and high pH. At low pH the situation is complex with deviations from Michaelis-Menten kinetics. Chloroplast carbonic anhydrase from higher plants have been reported to have primary structures that are completely different from the enzyme from animals. In addition, we find the circular dichroic spectrum of pea carbonic anhydrase to be well distinguished from that of human carbonic anhydrase II. Despite those structural differences the kinetic parameters indicate that pea carbonic anhydrase is equally efficient as human carbonic anhydrase II in catalysing the hydration of CO2. However, the mechanism for proton transfer from the active site to the surrounding medium seems to differ between the two enzymes.

  19. 21 CFR 158.170 - Frozen peas.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... Millimeters Inch Extra small Up to 7.5 0.295 Very small Up to 8.2 .32 Small Up to 8.75 .34 Medium Up to 10.2... peas, i.e., yellow or white but edible peas; (ii) Not more than 10 percent by weight blemished peas, i... cotyledons and loose skins, but excluding entire intact peas with skins detached; (v) Not more than...

  20. 29 CFR 780.139 - Pea vining.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 29 Labor 3 2010-07-01 2010-07-01 false Pea vining. 780.139 Section 780.139 Labor Regulations... âsuch Farming Operationâ-of the Farmer § 780.139 Pea vining. Vining employees of a pea vinery located on a farm, who vine only the peas grown on that particular farm, are engaged in agriculture. If they...

  1. 29 CFR 780.139 - Pea vining.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 29 Labor 3 2013-07-01 2013-07-01 false Pea vining. 780.139 Section 780.139 Labor Regulations... âsuch Farming Operationâ-of the Farmer § 780.139 Pea vining. Vining employees of a pea vinery located on a farm, who vine only the peas grown on that particular farm, are engaged in agriculture. If they...

  2. 29 CFR 780.139 - Pea vining.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 29 Labor 3 2012-07-01 2012-07-01 false Pea vining. 780.139 Section 780.139 Labor Regulations... âsuch Farming Operationâ-of the Farmer § 780.139 Pea vining. Vining employees of a pea vinery located on a farm, who vine only the peas grown on that particular farm, are engaged in agriculture. If they...

  3. 29 CFR 780.139 - Pea vining.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 29 Labor 3 2011-07-01 2011-07-01 false Pea vining. 780.139 Section 780.139 Labor Regulations... âsuch Farming Operationâ-of the Farmer § 780.139 Pea vining. Vining employees of a pea vinery located on a farm, who vine only the peas grown on that particular farm, are engaged in agriculture. If they...

  4. 29 CFR 780.139 - Pea vining.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 29 Labor 3 2014-07-01 2014-07-01 false Pea vining. 780.139 Section 780.139 Labor Regulations... âsuch Farming Operationâ-of the Farmer § 780.139 Pea vining. Vining employees of a pea vinery located on a farm, who vine only the peas grown on that particular farm, are engaged in agriculture. If they...

  5. 21 CFR 158.170 - Frozen peas.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 2 2014-04-01 2014-04-01 false Frozen peas. 158.170 Section 158.170 Food and... CONSUMPTION FROZEN VEGETABLES Requirements for Specific Standardized Frozen Vegetables § 158.170 Frozen peas. (a) Identity—(1) Product definition. Frozen peas is the food in “package” form as that term...

  6. Effectiveness of rhizobacteria containing ACC deaminase for growth promotion of peas (Pisum sativum) under drought conditions.

    PubMed

    Zahir, Z A; Munir, A; Asghar, H N; Shaharoona, B; Arshad, M

    2008-05-01

    A series of experiments were conducted to assess the effectiveness of rhizobacteria containing 1-aminocyclopropane- 1-carboxylate (ACC) deaminase for growth promotion of peas under drought conditions. Ten rhizobacteria isolated from the rhizosphere of different crops (peas, wheat, and maize) were screened for their growth promoting ability in peas under axenic condition. Three rhizobacterial isolates, Pseudomonas fluorescens biotype G (ACC-5), P. fluorescens (ACC-14), and P. putida biotype A (Q-7), were selected for pot trial on the basis of their source, ACC deaminase activity, root colonization, and growth promoting activity under axenic conditions. Inoculated and uninoculated (control) seeds of pea cultivar 2000 were sown in pots (4 seeds/pot) at different soil moisture levels (25, 50, 75, and 100% of field capacity). Results revealed that decreasing the soil moisture levels from 100 to 25% of field capacity significantly decreased the growth of peas. However, inoculation of peas with rhizobacteria containing ACC deaminase significantly decreased the "drought stress imposed effects" on growth of peas, although with variable efficacy at different moisture levels. At the lowest soil moisture level (25% field capacity), rhizobacterial isolate Pseudomonas fluorescens biotype G (ACC-5) was found to be more promising compared with the other isolates, as it caused maximum increases in fresh weight, dry weight, root length, shoot length, number of leaves per plant, and water use efficiency on fresh and dry weight basis (45, 150, 92, 45, 140, 46, and 147%, respectively) compared with respective uninoculated controls. It is highly likely that rhizobacteria containing ACC deaminase might have decreased the drought-stress induced ethylene in inoculated plants, which resulted in better growth of plants even at low moisture levels. Therefore, inoculation with rhizobacteria containing ACC deaminase could be helpful in eliminating the inhibitory effects of drought stress on the

  7. Characterization of proanthocyanidin metabolism in pea (Pisum sativum) seeds.

    PubMed

    Ferraro, Kiva; Jin, Alena L; Nguyen, Trinh-Don; Reinecke, Dennis M; Ozga, Jocelyn A; Ro, Dae-Kyun

    2014-09-16

    Proanthocyanidins (PAs) accumulate in the seeds, fruits and leaves of various plant species including the seed coats of pea (Pisum sativum), an important food crop. PAs have been implicated in human health, but molecular and biochemical characterization of pea PA biosynthesis has not been established to date, and detailed pea PA chemical composition has not been extensively studied. PAs were localized to the ground parenchyma and epidermal cells of pea seed coats. Chemical analyses of PAs from seeds of three pea cultivars demonstrated cultivar variation in PA composition. 'Courier' and 'Solido' PAs were primarily prodelphinidin-types, whereas the PAs from 'LAN3017' were mainly the procyanidin-type. The mean degree of polymerization of 'LAN3017' PAs was also higher than those from 'Courier' and 'Solido'. Next-generation sequencing of 'Courier' seed coat cDNA produced a seed coat-specific transcriptome. Three cDNAs encoding anthocyanidin reductase (PsANR), leucoanthocyanidin reductase (PsLAR), and dihydroflavonol reductase (PsDFR) were isolated. PsANR and PsLAR transcripts were most abundant earlier in seed coat development. This was followed by maximum PA accumulation in the seed coat. Recombinant PsANR enzyme efficiently synthesized all three cis-flavan-3-ols (gallocatechin, catechin, and afzalechin) with satisfactory kinetic properties. The synthesis rate of trans-flavan-3-ol by co-incubation of PsLAR and PsDFR was comparable to cis-flavan-3-ol synthesis rate by PsANR. Despite the competent PsLAR activity in vitro, expression of PsLAR driven by the Arabidopsis ANR promoter in wild-type and anr knock-out Arabidopsis backgrounds did not result in PA synthesis. Significant variation in seed coat PA composition was found within the pea cultivars, making pea an ideal system to explore PA biosynthesis. PsANR and PsLAR transcript profiles, PA localization, and PA accumulation patterns suggest that a pool of PA subunits are produced in specific seed coat cells early in

  8. Glycolate transporter of the pea chloroplast envelope

    SciTech Connect

    Howitz, K.T.

    1985-01-01

    The discovery of a glycolate transporter in the pea (Pisum sativum) chloroplast envelope is described. Several novel silicone oil centrifugation methods were developed to resolve the initial rate kinetics of (/sup 14/C)glycolate transport by isolated, intact pea chloroplasts. Chloroplast glycolate transport was found to be carrier mediated. Transport rates saturated with increasing glycolate concentration. N-Ethylmaleimide (NEM) pretreatment of chloroplasts inhibited transport, an inhibition prevented by glycolate. Glycolate distributed across the envelope in a way which equalized stromal and medium glycolic acid concentrations, limiting possible transport mechanisms to facilitated glycolic acid diffusion, proton symport or hydroxyl antiport. The effects of stomal and medium pH's on the K/sub m/ and V/sub max/ fit the predictions of mobile carrier kinetic models of hydroxyl antiport or proton symport (H/sup +/ binds first). The carrier mediated transport was fast enough to be consistent with in vivo rates of photorespiration. The 2-hydroxymonocarboxylates, glycerate, lactate and glyoxylate are competitive inhibitors of chloroplast glycolate uptake. Glyoxylate, D-lactate and D-glycerate cause glycolate counterflow, indicating that they are also substrates of the glycolate carrier. This finding was confirmed for D-glycerate by studies on glycolate effects on (1-/sup 14/C)D-glycerate transport.

  9. Molecular diversity of native rhizobia trapped by five field pea genotypes in Indian soils.

    PubMed

    Wadhwa, K; Dudeja, S S; Yadav, R K

    2011-02-01

    Five pea cultivars; HFP 4, HVP 3-5, HFP 9426, Jayanti and Hariyal, being grown in CCS Haryana Agricultural University farm were used to isolate native rhizobia. Selected 54 rhizobia, from all cultivars, were authenticated as rhizobia by plant infectivity test. Along with nodulation, symbiotic effectiveness in terms of symbiotic ratios showed wide range of effectiveness of pea rhizobia from 1.11 to 5.0. DNA of all the 54 rhizobia was extracted and amplified by PCR, using ERIC and 16S rDNA primers. Dendrogram based on ERIC profiles of these 54 rhizobia showed the formation of 13 subclusters at 80% level of similarity. Dendrogram based on RFLP of 16S rDNA by three restriction endonucleases; Msp I, Csp 6I and Rsa I; also formed 13 subclusters at 80% level of similarity. However, positioning of subclusters was different from that of ERIC based dendrogram. Majority of the isolates i.e. 64.8% by ERIC profiles and 44.4% by RFLP of 16S rDNA formed one cluster. Isolates from same nodule were not 100% similar. Considering each cluster representing a rhizobial genotype, both techniques used to assess molecular diversity indicated the presence of 13 genotypes of field pea rhizobia in CCS Haryana Agricultural University farm soil. Two pea rhizobial genotypes were able to nodulate all the five pea cultivars. Furthermore, high strain richness index (0.43-0.5) of field pea rhizobia was observed by both the techniques.

  10. Raised expression of the antiapoptotic protein ped/pea-15 increases susceptibility to chemically induced skin tumor development.

    PubMed

    Formisano, Pietro; Perruolo, Giuseppe; Libertini, Silvana; Santopietro, Stefania; Troncone, Giancarlo; Raciti, Gregory Alexander; Oriente, Francesco; Portella, Giuseppe; Miele, Claudia; Beguinot, Francesco

    2005-10-27

    ped/pea-15 is a cytosolic protein performing a broad antiapoptotic function. We show that, upon DMBA/TPA-induced skin carcinogenesis, transgenic mice overexpressing ped/pea-15 (Tg(ped/pea-15)) display early development of papillomas and a four-fold increase in papilloma number compared to the nontransgenic littermates (P<0.001). The malignant conversion frequency was 24% for the Tg(ped/pea-15) mice and only 5% in controls (P<0.01). The isolated application of TPA, but not that of DMBA, was sufficient to reversibly upregulate ped/pea-15 in both untransformed skin and cultured keratinocytes. ped/pea-15 protein levels were also increased in DMBA/TPA-induced papillomas of both Tg(ped/pea-15) and control mice. Isolated TPA applications induced Caspase-3 activation and apoptosis in nontransformed mouse epidermal tissues. The induction of both Caspase-3 and apoptosis by TPA were four-fold inhibited in the skin of the Tg(ped/pea-15) compared to the nontransgenic mice, accompanied by a similarly sized reduction in TPA-induced JNK and p38 stimulation and by constitutive induction of cytoplasmic ERK activity in the transgenics. ped/pea-15 expression was stably increased in cell lines from DMBA/TPA-induced skin papillomas and carcinomas, paralleled by protection from TPA apoptosis. In the A5 spindle carcinoma cell line, antisense inhibition of ped/pea-15 expression simultaneously rescued sensitivity to TPA-induced Caspase-3 function and apoptosis. The antisense also reduced A5 cell ability to grow in semisolid media by 65% (P<0.001) and increased by three-fold tumor latency time (P<0.01). Thus, the expression levels of ped/pea-15 control Caspase-3 function and epidermal cell apoptosis in vivo and determine susceptibility to skin tumor development.

  11. Multilocus Sequence Typing Confirms Wild Birds as the Source of a Campylobacter Outbreak Associated with the Consumption of Raw Peas

    PubMed Central

    Xavier, Catherine; Santovenia, Monica; Pruckler, Janet; Stroika, Steven; Joyce, Kevin; Gardner, Tracie; Fields, Patricia I.; McLaughlin, Joe; Tauxe, Robert V.; Fitzgerald, Collette

    2014-01-01

    From August to September 2008, the Centers for Disease Control and Prevention (CDC) assisted the Alaska Division of Public Health with an outbreak investigation of campylobacteriosis occurring among the residents of Southcentral Alaska. During the investigation, pulsed-field gel electrophoresis (PFGE) of Campylobacter jejuni isolates from human, raw pea, and wild bird fecal samples confirmed the epidemiologic link between illness and the consumption of raw peas contaminated by sandhill cranes for 15 of 43 epidemiologically linked human isolates. However, an association between the remaining epidemiologically linked human infections and the pea and wild bird isolates was not established. To better understand the molecular epidemiology of the outbreak, C. jejuni isolates (n = 130; 59 from humans, 40 from peas, and 31 from wild birds) were further characterized by multilocus sequence typing (MLST). Here we present the molecular evidence to demonstrate the association of many more human C. jejuni infections associated with the outbreak with raw peas and wild bird feces. Among all sequence types (STs) identified, 26 of 39 (67%) were novel and exclusive to the outbreak. Five clusters of overlapping STs (n = 32 isolates; 17 from humans, 2 from peas, and 13 from wild birds) were identified. In particular, cluster E (n = 7 isolates; ST-5049) consisted of isolates from humans, peas, and wild birds. Novel STs clustered closely with isolates typically associated with wild birds and the environment but distinct from lineages commonly seen in human infections. Novel STs and alleles recovered from human outbreak isolates allowed additional infections caused by these rare genotypes to be attributed to the contaminated raw peas. PMID:24837383

  12. Multilocus sequence typing confirms wild birds as the source of a Campylobacter outbreak associated with the consumption of raw peas.

    PubMed

    Kwan, Patrick S L; Xavier, Catherine; Santovenia, Monica; Pruckler, Janet; Stroika, Steven; Joyce, Kevin; Gardner, Tracie; Fields, Patricia I; McLaughlin, Joe; Tauxe, Robert V; Fitzgerald, Collette

    2014-08-01

    From August to September 2008, the Centers for Disease Control and Prevention (CDC) assisted the Alaska Division of Public Health with an outbreak investigation of campylobacteriosis occurring among the residents of Southcentral Alaska. During the investigation, pulsed-field gel electrophoresis (PFGE) of Campylobacter jejuni isolates from human, raw pea, and wild bird fecal samples confirmed the epidemiologic link between illness and the consumption of raw peas contaminated by sandhill cranes for 15 of 43 epidemiologically linked human isolates. However, an association between the remaining epidemiologically linked human infections and the pea and wild bird isolates was not established. To better understand the molecular epidemiology of the outbreak, C. jejuni isolates (n=130; 59 from humans, 40 from peas, and 31 from wild birds) were further characterized by multilocus sequence typing (MLST). Here we present the molecular evidence to demonstrate the association of many more human C.jejuni infections associated with the outbreak with raw peas and wild bird feces. Among all sequence types (STs) identified, 26 of 39 (67%) were novel and exclusive to the outbreak. Five clusters of overlapping STs (n=32 isolates; 17 from humans, 2 from peas, and 13 from wild birds) were identified. In particular, cluster E (n=7 isolates; ST-5049) consisted of isolates from humans,peas, and wild birds. Novel STs clustered closely with isolates typically associated with wild birds and the environment but distinct from lineages commonly seen in human infections. Novel STs and alleles recovered from human outbreak isolates allowed additional infections caused by these rare genotypes to be attributed to the contaminated raw peas.

  13. Superdeformed nuclei

    SciTech Connect

    Janssens, R.V.F.; Khoo, Teng Lek.

    1991-01-01

    This paper reviews the most recent advances in the understanding of the physics of superdeformed nuclei from the point of view of the experimentalists. It covers among other subjects the following topics: (1) the discovery of a new region of superdeformed nuclei near A=190, (2) the surprising result of the occurrence of bands with identical transition energies in neighboring superdeformed nuclei near A=150 and A=190, (3) the importance of octupole degrees of freedom at large deformation and (4) the properties associated with the feeding and the decay of superdeformed bands. The text presented hereafter will appear as a contribution to the Annual Review of Nuclear and Particle Science, Volume 41. 88 refs., 11 figs.

  14. Cosmogenic nuclei

    NASA Technical Reports Server (NTRS)

    Raisbeck, G. M.

    1986-01-01

    Cosmogenic nuclei, nuclides formed by nuclear interactions of galactic and solar cosmic rays with extraterrestrial or terrestrial matter are discussed. Long lived radioactive cosmogenic isotopes are focused upon. Their uses in dating, as tracers of the interactions of cosmic rays with matter, and in obtaining information on the variation of primary cosmic ray flux in the past are discussed.

  15. Twins born in different environments? Nuclei of two dSphs: isolated galaxy KKS3 and E269-66, a close neighbor of NGC5128

    NASA Astrophysics Data System (ADS)

    Sharina, Margarita; Kniazev, Alexei; Karachentsev, Igor

    2017-03-01

    We present the results of age, metallicity and radial velocity determination for central massive globular clusters (GCs) in dwarf spheroidal galaxies: KKs3 and ESO269-66. KKS3 is a unique isolated galaxy. ESO269-66 is a close neighbor of the giant S0 Centaurus A. The results contribute to the knowledge about the origin of massive star clusters and their host dSphs. The structure and star formation histories of the two dwarf galaxies look rather similar. Both of them have experienced several star-forming events. The most recent ones occurred 1-2 Gyr ago, and most powerful bursts happened 12-14 Gyrs ago. Our analysis has shown that both GCs appear to be 1-2 Gyr younger and 0.1-0.3 dex more metal-rich than the most ancient metal-poor stars in the host dSphs. We examine signatures of multiple stellar population in the GCs using our data. Since central star-forming bursts were extended in time, the massive clusters might be considered as nuclei of the galaxies.

  16. Nodulation, Nitrogen Fixation, and Hydrogen Oxidation by Pigeon Pea Bradyrhizobium spp. in Symbiotic Association with Pigeon Pea, Cowpea, and Soybean †

    PubMed Central

    Nautiyal, C. S.; Hegde, S. V.; van Berkum, P.

    1988-01-01

    The pigeon pea strains of Bradyrhizobium CC-1, CC-8, UASGR(S), and F4 were evaluated for nodulation, effectiveness for N2 fixation, and H2 oxidation with homologous and nonhomologous host plants. Strain CC-1 nodulated Macroptilium atropurpureum, Vigna unguiculata, Glycine max, and G. soja but did not nodulate Pisum sativum, Phaseolus vulgaris, Trigonella foenum-graecum, and Trifolium repens. Strain F4 nodulated G. max cv. Peking and PI 434937 (Malayan), but the symbioses formed were poor. Similarly, G. max cv. Peking, cv. Bragg, PI 434937, PR 13-28-2-8-7, and HM-1 were nodulated by strain CC-1, and symbioses were also poor. G. max cv. Williams and cv. Clark were not nodulated. H2 uptake activity was expressed with pigeon pea and cowpea, but not with soybean. G. max cv. Bragg grown in Bangalore, India, in local soil not previously exposed to Bradyrhizobium japonicum formed nodules with indigenous Bradyrhizobium spp. Six randomly chosen isolates, each originating from a different nodule, formed effective symbioses with pigeon pea host ICPL-407, nodulated PR 13-28-2-8-7 soybean forming moderately effective symbioses, and did not nodulate Williams soybean. These results indicate the six isolates to be pigeon pea strains although they originated from soybean nodules. Host-determined nodulation of soybean by pigeon pea Bradyrhizobium spp. may depend upon the ancestral backgrounds of the cultivars. The poor symbioses formed by the pigeon pea strains with soybean indicate that this crop should be inoculated with B. japonicum for its cultivation in soils containing only pigeon pea Bradyrhizobium spp. PMID:16347542

  17. Exotic atomic nuclei

    NASA Astrophysics Data System (ADS)

    Hamilton, J. H.; Maruhn, J. A.

    1986-07-01

    From the study of nuclei with abundances of neutrons and protons (N numbers and Z numbers) quite different from those found in nature, it has been possible to gain new views of motions and structures within nuclear matter. Based on the spherical shell model of the nucleus proposed by Mayer and Jensen in 1949 and the collective model of nuclear deformation proposed in 1952 by Bohr and Mottelson, it has come to be possible to decide what shape or shapes a nucleus must have for a given set of N and Z numbers. It turns out that not only spherical nuclei are possible but also prolate and oblate spheroids (football and discus shaped), triaxial (like a partially deflated football), and even pear- or peanut-shaped. A significant experimental tool in such studies is the ISOL or Isotope-Separator, On-Line, which makes possible the construction of energy level diagrams from the study of exotic nuclei created when particles from accelerators strike various kinds of foil. The significance of magic numbers and super-magic numbers (particular combinations of N and Z) for the stability of various exotic nuclei is considered. International facilities engaged in such studies are noted.

  18. Developmental differences in posttranslational calmodulin methylation in pea plants

    SciTech Connect

    Oh, Sukheung; Roberts, D.M. )

    1990-05-01

    A calmodulin-N-methyltransferase was used to analyze the degree of lysine-115 methylation of pea calmodulin. Calmodulin was isolated from segments of developing roots of young etiolated and green pea plants and was tested for its ability to be methylated by the calmodulin methyltransferase in the presence of {sup 3}H-methyl-S-adenosylmethionine. Calmodulin methylation levels were lower in apical root segments and in the young lateral roots compared with the mature, differentiated root tissues. The methylation of these calmodulin samples occurs specifically at lysine 115 since site-directed mutants of calmodulin with substitutions at this position were not methylated and competitively inhibited methylation. The present findings, combined with previous data showing differences in NAD kinase activation by methylated and unmethylated calmodulins, raise the possibility that posttranslational methylation could affect calmodulin action.

  19. Strigolactones promote nodulation in pea.

    PubMed

    Foo, Eloise; Davies, Noel W

    2011-11-01

    Strigolactones are recently defined plant hormones with roles in mycorrhizal symbiosis and shoot and root architecture. Their potential role in controlling nodulation, the related symbiosis between legumes and Rhizobium bacteria, was explored using the strigolactone-deficient rms1 mutant in pea (Pisum sativum L.). This work indicates that endogenous strigolactones are positive regulators of nodulation in pea, required for optimal nodule number but not for nodule formation per se. rms1 mutant root exudates and root tissue are almost completely deficient in strigolactones, and rms1 mutant plants have approximately 40% fewer nodules than wild-type plants. Treatment with the synthetic strigolactone GR24 elevated nodule number in wild-type pea plants and also elevated nodule number in rms1 mutant plants to a level similar to that seen in untreated wild-type plants. Grafting studies revealed that nodule number and strigolactone levels in root tissue of rms1 roots were unaffected by grafting to wild-type scions indicating that strigolactones in the root, but not shoot-derived factors, regulate nodule number and provide the first direct evidence that the shoot does not make a major contribution to root strigolactone levels.

  20. Physics of Unstable Nuclei

    NASA Astrophysics Data System (ADS)

    Khoa, Dao Tien; Egelhof, Peter; Gales, Sydney; Giai, Nguyen Van; Motobayashi, Tohru

    2008-04-01

    ]C([symbol], n)[symbol]O by the transfer reaction [symbol]C([symbol]Li, t)[symbol]O / F. Hammache et al. -- SPIRAL2 at GANIL: a world of leading ISOL facility for the physics of exotic nuclei / S. Gales -- Magnetic properties of light neutron-rich nuclei and shell evolution / T. Suzuki, T. Otsuka -- Multiple scattering effects in elastic and quasi free proton scattering from halo nuclei / R. Crespo et al. -- The dipole response of neutron halos and skins / T. Aumann -- Giant and pygmy resonances within axially-symmetric-deformed QRPA with the Gogny force / S. Péru, H. Goutte -- Soft K[symbol] = O+ modes unique to deformed neutron-rich unstable nuclei / K. Yoshida et al. -- Synthesis, decay properties, and identification of superheavy nuclei produced in [symbol]Ca-induced reactions / Yu. Ts. Oganessian et al. -- Highlights of the Brazilian RIB facility and its first results and hindrance of fusion cross section induced by [symbol]He / P. R. S. Gomes et al. -- Search for long fission times of super-heavy elements with Z = 114 / M. Morjean et al. -- Microscopic dynamics of shape coexistence phenomena around [symbol]Se and [symbol]Kr / N. Hinohara et al. -- [symbol]-cluster states and 4[symbol]-particle condensation in [symbol]O / Y. Funaki et al. -- Evolution of the N = 28 shell closure far from stability / O. Sorlin et al. -- Continuum QRPA approach and the surface di-neutron modes in nuclei near the neutron drip-line / M. Matsuo et al. -- Deformed relativistic Hartree-Bogoliubov model for exotic nuclei / S. G. Zhou et al. -- Two- and three-body correlations in three-body resonances and continuum states / K. Katō, K. Ikeda -- Pion- and Rho-Meson effects in relativistic Hartree-Fock and RPA / N. V. Giai et al. -- Study of the structure of neutron rich nuclei by using [symbol]-delayed neutron and gamma emission method / Y. Ye et al. -- Production of secondary radioactive [symbol] Na beam for the study of [symbol]Na([symbol], p)[symbol]Mg stellar reaction / D. N. Binh et al

  1. Discrete forms of amylose are synthesized by isoforms of GBSSI in pea.

    PubMed

    Edwards, Anne; Vincken, Jean-Paul; Suurs, Luc C J M; Visser, Richard G F; Zeeman, Sam; Smith, Alison; Martin, Cathie

    2002-08-01

    Amyloses with distinct molecular masses are found in the starch of pea embryos compared with the starch of pea leaves. In pea embryos, a granule-bound starch synthase protein (GBSSIa) is required for the synthesis of a significant portion of the amylose. However, this protein seems to be insignificant in the synthesis of amylose in pea leaves. cDNA clones encoding a second isoform of GBSSI, GBSSIb, have been isolated from pea leaves. Comparison of GBSSIa and GBSSIb activities shows them to have distinct properties. These differences have been confirmed by the expression of GBSSIa and GBSSIb in the amylose-free mutant of potato. GBSSIa and GBSSIb make distinct forms of amylose that differ in their molecular mass. These differences in product specificity, coupled with differences in the tissues in which GBSSIa and GBSSIb are most active, explain the distinct forms of amylose found in different tissues of pea. The shorter form of amylose formed by GBSSIa confers less susceptibility to the retrogradation of starch pastes than the amylose formed by GBSSIb. The product specificity of GBSSIa could provide beneficial attributes to starches for food and nonfood uses.

  2. The enhanced rate of transcription of methyl mercury-exposed DNA by RNA polymerase is not sufficient to explain the stimulatory effect of methyl mercury on RNA synthesis in isolated nuclei.

    PubMed

    Frenkel, G D; Ducote, J

    1987-10-01

    Previous work demonstrated two stimulatory effects of methyl mercury on nucleic acid synthesis: (1) in isolated nuclei, methyl mercury stimulates RNA synthesis which is catalyzed by RNA polymerase II [Frenkel and Randles, J. Biol. Chem. 257, 6275-6279 (1982)]. (2) Brief exposure of purified DNA to methyl mercury increases the rate of its transcription by purified RNA polymerase II [Frenkel, Cain, and Chao, Biochem. Biophys. Res. Commun. 127, 849-856 (1985)]. The latter effect was considered as a possible mechanism of the former. Two lines of evidence are presented here which demonstrate that the latter effect is not a sufficient explanation for the former. (1) Mercuric perchlorate has been found to increase the rate of DNA transcription by purified polymerase and the template properties of the mercuric perchlorate-exposed DNA have been found to resemble those of methyl mercury-exposed DNA. Nevertheless, mercuric perchlorate has been shown not to stimulate RNA synthesis in isolated HeLa nuclei. (2) In isolated nuclei of the B50 rat neuroblastoma cell line, RNA synthesis has been found to be stimulated only minimally by methyl mercury. Nevertheless, RNA polymerase II purified from the B50 cells has been found to transcribe methyl mercury-exposed DNA at a higher rate than unexposed control DNA.

  3. Twins born in different environments? Nuclei of two dSphs: isolated galaxy KKS3 and ESO269-66, a close neighbor of NGC5128

    NASA Astrophysics Data System (ADS)

    Sharina, Margarita; Karachentsev, Igor; Kniazev, Alexei

    2015-08-01

    The close vicinity of giant neighbors determines the environmental mechanisms that have been considered responsible for the evolution of dwarf spheroidal galaxies (dSphs). In the recent years, Karachentsev and collaborators have reported on the discovery of a few truly isolated dSphs in the Local volume. This study focuses on one of these unusual objects, KKs3 (MV=-12.3 mag). It contains a massive globular cluster (GC) (MV=-8.5 mag) near its optical center. We have performed the estimation of its radial velocity using a medium-resolution spectrum obtained with the RSS spectrograph at the Southern African Large Telescope (SALT). The signal-to-noise ratio in the spectrum was sufficient to estimate the age and metallicity for the GC using simple stellar population models, and the methods of full spectrum fitting and Lick index diagnostic diagrams. The results contribute to the knowledge about the origin of massive star clusters and their host dSphs.In the same way we have analyzed another luminous GC (MV=-10) in the center of ESO269-66 (MB=-15.4), a close dSph neighbor of the giant S0 Cen A. The cluster was observed with SALT in the same instrumental configuration. The structure and star formation histories of the two galaxies look rather similar. Both of them have experienced several star-forming events. The most recent ones occurred 1÷2 Gyr ago, and most powerful bursts happened 12÷14 Gyrs ago. Our analysis has shown that both GCs appear to be 1÷2 Gyr younger and 0.2÷0.3 dex more metal-rich than the most ancient metal-poor stars in the host dSphs. We examine signatures of multiple stellar population in the GCs using out data. Since central star-forming bursts were extended in time, the massive clusters might be considered as nuclei of the galaxies.

  4. EDTA a novel inducer of pisatin, a phytoalexin indicator of the non-host resistance in peas.

    PubMed

    Hadwiger, Lee A; Tanaka, Kiwamu

    2014-12-23

    Pea pod endocarp suppresses the growth of an inappropriate fungus or non-pathogen by generating a "non-host resistance response" that completely suppresses growth of the challenging fungus within 6 h. Most of the components of this resistance response including pisatin production can be elicited by an extensive number of both biotic and abiotic inducers. Thus this phytoalexin serves as an indicator to be used in evaluating the chemical properties of inducers that can initiate the resistance response. Many of the pisatin inducers are reported to interact with DNA and potentially cause DNA damage. Here we propose that EDTA (ethylenediaminetetraacetic acid) is an elicitor to evoke non-host resistance in plants. EDTA is manufactured as a chelating agent, however at low concentration it is a strong elicitor, inducing the phytoalexin pisatin, cellular DNA damage and defense-responsive genes. It is capable of activating complete resistance in peas against a pea pathogen. Since there is also an accompanying fragmentation of pea DNA and alteration in the size of pea nuclei, the potential biochemical insult as a metal chelator may not be its primary action. The potential effects of EDTA on the structure of DNA within pea chromatin may assist the transcription of plant defense genes.

  5. Exotic Nuclei

    SciTech Connect

    Galindo-Uribarri, Alfredo {nmn}

    2010-01-01

    Current experimental developments on the study of exotic nuclei far from the valley of stability are discussed. I start with general aspects related to the production of radioactive beams followed by the description of some of the experimental tools and specialized techniques for studies in reaction spectroscopy, nuclear structure research and nuclear applications with examples from selected topical areas with which I have been involved. I discuss some of the common challenges faced in Accelerator Mass Spectrometry (AMS) and Radioactive Ion Beam (RIB) science.

  6. Nuclei and Fundamental Symmetries

    NASA Astrophysics Data System (ADS)

    Haxton, Wick

    2016-09-01

    Nuclei provide marvelous laboratories for testing fundamental interactions, often enhancing weak processes through accidental degeneracies among states, and providing selection rules that can be exploited to isolate selected interactions. I will give an overview of current work, including the use of parity violation to probe unknown aspects of the hadronic weak interaction; nuclear electric dipole moment searches that may shed light on new sources of CP violation; and tests of lepton number violation made possible by the fact that many nuclei can only decay by rare second-order weak interactions. I will point to opportunities in both theory and experiment to advance the field. Based upon work supported in part by the US Department of Energy, Office of Science, Office of Nuclear Physics and SciDAC under Awards DE-SC00046548 (Berkeley), DE-AC02-05CH11231 (LBNL), and KB0301052 (LBNL).

  7. Yield potential of pigeon pea cultivars

    USDA-ARS?s Scientific Manuscript database

    Yield potential of twelve vegetable pigeon pea (Cajanus cajun) cultivars was evaluated at two locations in eastern Kenya during 2012 and 2013 cropping years. Pigeon pea pod numbers, seeds per pod, seed mass, grain yield and shelling percentage were quantified in three replicated plots, arranged in a...

  8. Growth parameters of vegetable pigeon pea cultivars

    USDA-ARS?s Scientific Manuscript database

    Pigeon pea is an important crop in the dry regions of eastern Kenya, due to its drought tolerance and high protein content; however, farmer’s yield is limiting. Ojwang et al. (HortTech Vol 26 (1), 2016) evaluated twelve pigeon pea cultivars for flowering, plant height, branches, pod length and yield...

  9. 21 CFR 155.170 - Canned peas.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 2 2013-04-01 2013-04-01 false Canned peas. 155.170 Section 155.170 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN.... (a) Identity—(1) Definition. Canned peas is the food prepared from fresh or frozen succulent seeds of...

  10. 21 CFR 155.170 - Canned peas.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 2 2012-04-01 2012-04-01 false Canned peas. 155.170 Section 155.170 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN.... (a) Identity—(1) Definition. Canned peas is the food prepared from fresh or frozen succulent seeds of...

  11. Immunity and other defenses in pea aphids, Acyrthosiphon pisum

    PubMed Central

    2010-01-01

    Background Recent genomic analyses of arthropod defense mechanisms suggest conservation of key elements underlying responses to pathogens, parasites and stresses. At the center of pathogen-induced immune responses are signaling pathways triggered by the recognition of fungal, bacterial and viral signatures. These pathways result in the production of response molecules, such as antimicrobial peptides and lysozymes, which degrade or destroy invaders. Using the recently sequenced genome of the pea aphid (Acyrthosiphon pisum), we conducted the first extensive annotation of the immune and stress gene repertoire of a hemipterous insect, which is phylogenetically distantly related to previously characterized insects models. Results Strikingly, pea aphids appear to be missing genes present in insect genomes characterized to date and thought critical for recognition, signaling and killing of microbes. In line with results of gene annotation, experimental analyses designed to characterize immune response through the isolation of RNA transcripts and proteins from immune-challenged pea aphids uncovered few immune-related products. Gene expression studies, however, indicated some expression of immune and stress-related genes. Conclusions The absence of genes suspected to be essential for the insect immune response suggests that the traditional view of insect immunity may not be as broadly applicable as once thought. The limitations of the aphid immune system may be representative of a broad range of insects, or may be aphid specific. We suggest that several aspects of the aphid life style, such as their association with microbial symbionts, could facilitate survival without strong immune protection. PMID:20178569

  12. Effective stabilization of CLA by microencapsulation in pea protein.

    PubMed

    Costa, A M M; Nunes, J C; Lima, B N B; Pedrosa, C; Calado, V; Torres, A G; Pierucci, A P T R

    2015-02-01

    CLA was microencapsulated by spray drying in ten varied wall systems (WS) consisting of pea protein isolate or pea protein concentrate (PPC) alone at varied core:WS ratios (1:2; 1:3 and 1:4), or blended with maltodextrin (M) and carboxymethylcellulose at a pea protein:carbohydrate ratio of 3:1. The physical-chemical properties of the CLA microparticles were characterised by core retention, microencapsulation efficiency (ME), particle size and moisture. CLA:M:PPC (1:1:3) showed the most promising results, thus we evaluated the effect of M addition in the WS on other physical-chemical characteristics and oxidative stability (CLA isomer profile, quantification of CLA and volatile compounds by SPME coupled with CG-MS) during two months of storage at room temperature, CLA:PPC (1:4) was selected for comparisons. CLA:M:PPC (1:1:3) microparticles demonstrated better morphology, solubility, dispersibility and higher glass-transition temperature values. M addition did not influence the oxidative stability of CLA, however its presence improved physical-chemical characteristics necessary for food applications.

  13. Characterization of Two Brassinosteroid C-6 Oxidase Genes in Pea1[W][OA

    PubMed Central

    Jager, Corinne E.; Symons, Gregory M.; Nomura, Takahito; Yamada, Yumiko; Smith, Jennifer J.; Yamaguchi, Shinjiro; Kamiya, Yuji; Weller, James L.; Yokota, Takao; Reid, James B.

    2007-01-01

    C-6 oxidation genes play a key role in the regulation of biologically active brassinosteroid (BR) levels in the plant. They control BR activation, which involves the C-6 oxidation of 6-deoxocastasterone (6-DeoxoCS) to castasterone (CS) and in some cases the further conversion of CS to brassinolide (BL). C-6 oxidation is controlled by the CYP85A family of cytochrome P450s, and to date, two CYP85As have been isolated in tomato (Solanum lycopersicum), two in Arabidopsis (Arabidopsis thaliana), one in rice (Oryza sativa), and one in grape (Vitis vinifera). We have now isolated two CYP85As (CYP85A1 and CYP85A6) from pea (Pisum sativum). However, unlike Arabidopsis and tomato, which both contain one BR C-6 oxidase that converts 6-DeoxoCS to CS and one BR C-6 Baeyer-Villiger oxidase that converts 6-DeoxoCS right through to BL, the two BR C-6 oxidases in pea both act principally to convert 6-DeoxoCS to CS. The isolation of these two BR C-6 oxidation genes in pea highlights the species-specific differences associated with C-6 oxidation. In addition, we have isolated a novel BR-deficient mutant, lke, which blocks the function of one of these two BR C-6 oxidases (CYP85A6). The lke mutant exhibits a phenotype intermediate between wild-type plants and previously characterized pea BR mutants (lk, lka, and lkb) and contains reduced levels of CS and increased levels of 6-DeoxoCS. To date, lke is the only mutant identified in pea that blocks the latter steps of BR biosynthesis and it will therefore provide an excellent tool to further examine the regulation of BR biosynthesis and the relative biological activities of CS and BL in pea. PMID:17322341

  14. Spinocerebellar ataxias types 2 and 3: degeneration of the pre-cerebellar nuclei isolates the three phylogenetically defined regions of the cerebellum.

    PubMed

    Rüb, U; Gierga, K; Brunt, E R; de Vos, R A I; Bauer, M; Schöls, L; Bürk, K; Auburger, G; Bohl, J; Schultz, C; Vuksic, M; Burbach, G J; Braak, H; Deller, T

    2005-11-01

    The pre-cerebellar nuclei act as a gate for the entire neocortical, brainstem and spinal cord afferent input destined for the cerebellum. Since no pathoanatomical studies of these nuclei had yet been performed in spinocerebellar ataxia type 2 (SCA2) or type 3 (SCA3), we carried out a detailed postmortem study of the pre-cerebellar nuclei in six SCA2 and seven SCA3 patients in order to further characterize the extent of brainstem degeneration in these ataxic disorders. By means of unconventionally thick serial sections through the brainstem stained for lipofuscin pigment and Nissl material, we could show that all of the pre-cerebellar nuclei (red, pontine, arcuate, prepositus hypoglossal, superior vestibular, lateral vestibular, medial vestibular, interstitial vestibular, spinal vestibular, vermiform, lateral reticular, external cuneate, subventricular, paramedian reticular, intercalate, interfascicular hypoglossal, and conterminal nuclei, pontobulbar body, reticulotegmental nucleus of the pons, inferior olive, and nucleus of Roller) are among the targets of both of the degenerative processes underlying SCA2 and SCA3. These novel findings are in contrast to the current neuropathological literature, which assumes that only a subset of pre-cerebellar nuclei in SCA2 and SCA3 may undergo neurodegeneration. Widespread damage to the pre-cerebellar nuclei separates all three phylogenetically and functionally defined regions of the cerebellum, impairs their physiological functions and thus explains the occurrence of gait, stance, limb and truncal ataxia, dysarthria, truncal and postural instability with disequilibrium, impairments of the vestibulo-ocular reaction and optokinetic nystagmus, slowed and saccadic smooth pursuits, dysmetrical horizontal saccades, and gaze-evoked nystagmus during SCA2 and SCA3.

  15. Pea amyloplast DNA is qualitatively similar to pea chloroplast DNA

    NASA Technical Reports Server (NTRS)

    Gaynor, J. J.

    1984-01-01

    Amyloplast DNA (apDNA), when subjected to digestion with restriction endonucleases, yields patterns nearly identical to that of DNA from mature pea chloroplasts (ctDNA). Southern transfers of apDNA and ctDNA, probed with the large subunit (LS) gene of ribulose-1,5-bisphosphate carboxylase (Rubisco), shows hybridization to the expected restriction fragments for both apDNA and ctDNA. However, Northern transfers of total RNA from chloroplasts and amyloplasts, probed again with the LS gene of Rubisco, shows that no detectable LS meggage is found in amyloplasts although LS expression in mature chloroplasts is high. Likewise, two dimensional polyacrylamide gel electrophoresis of etiolated gravisensitive pea tissue shows that both large and small subunits of Rubisco are conspicuously absent; however, in greening tissue these two constitute the major soluble proteins. These findings suggest that although the informational content of these two organelle types is equivalent, gene expression is quite different and is presumably under nuclear control.

  16. Pea amyloplast DNA is qualitatively similar to pea chloroplast DNA

    NASA Technical Reports Server (NTRS)

    Gaynor, J. J.

    1984-01-01

    Amyloplast DNA (apDNA), when subjected to digestion with restriction endonucleases, yields patterns nearly identical to that of DNA from mature pea chloroplasts (ctDNA). Southern transfers of apDNA and ctDNA, probed with the large subunit (LS) gene of ribulose-1,5-bisphosphate carboxylase (Rubisco), shows hybridization to the expected restriction fragments for both apDNA and ctDNA. However, Northern transfers of total RNA from chloroplasts and amyloplasts, probed again with the LS gene of Rubisco, shows that no detectable LS meggage is found in amyloplasts although LS expression in mature chloroplasts is high. Likewise, two dimensional polyacrylamide gel electrophoresis of etiolated gravisensitive pea tissue shows that both large and small subunits of Rubisco are conspicuously absent; however, in greening tissue these two constitute the major soluble proteins. These findings suggest that although the informational content of these two organelle types is equivalent, gene expression is quite different and is presumably under nuclear control.

  17. 7 CFR 457.137 - Green pea crop insurance provisions.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    .... Green peas genetically developed to be eaten without shelling (e.g., snap peas, snow peas, and Chinese... peas genetically developed to be shelled prior to eating, canning or freezing. 2. Unit Division (a) For..., either of which causes the acreage to be bypassed. (2) Fire; (3) Insects, but not damage due to...

  18. 21 CFR 155.172 - Canned dry peas.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Canned dry peas. 155.172 Section 155.172 Food and... peas. (a) Identity. Canned dry peas conforms to the definition and standard of identity, and is subject to the requirements for label declaration of ingredients, prescribed for canned peas by § 155.170(a...

  19. 21 CFR 155.172 - Canned dry peas.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 2 2014-04-01 2014-04-01 false Canned dry peas. 155.172 Section 155.172 Food and... peas. (a) Identity. Canned dry peas conforms to the definition and standard of identity, and is subject to the requirements for label declaration of ingredients, prescribed for canned peas by § 155.170(a...

  20. 21 CFR 155.172 - Canned dry peas.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 2 2011-04-01 2011-04-01 false Canned dry peas. 155.172 Section 155.172 Food and... peas. (a) Identity. Canned dry peas conforms to the definition and standard of identity, and is subject to the requirements for label declaration of ingredients, prescribed for canned peas by § 155.170(a...

  1. 21 CFR 155.172 - Canned dry peas.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 2 2013-04-01 2013-04-01 false Canned dry peas. 155.172 Section 155.172 Food and... peas. (a) Identity. Canned dry peas conforms to the definition and standard of identity, and is subject to the requirements for label declaration of ingredients, prescribed for canned peas by § 155.170(a...

  2. 21 CFR 155.172 - Canned dry peas.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 2 2012-04-01 2012-04-01 false Canned dry peas. 155.172 Section 155.172 Food and... peas. (a) Identity. Canned dry peas conforms to the definition and standard of identity, and is subject to the requirements for label declaration of ingredients, prescribed for canned peas by § 155.170(a...

  3. Mitochondrial and Nuclear Localization of a Novel Pea Thioredoxin: Identification of Its Mitochondrial Target Proteins1[W

    PubMed Central

    Martí, María C.; Olmos, Enrique; Calvete, Juan J.; Díaz, Isabel; Barranco-Medina, Sergio; Whelan, James; Lázaro, Juan J.; Sevilla, Francisca; Jiménez, Ana

    2009-01-01

    Plants contain several genes encoding thioredoxins (Trxs), small proteins involved in the regulation of the activity of many enzymes through dithiol-disulfide exchange. In addition to chloroplastic and cytoplasmic Trx systems, plant mitochondria contain a reduced nicotinamide adenine dinucleotide phosphate-dependent Trx reductase and a specific Trx o, and to date, there have been no reports of a gene encoding a plant nuclear Trx. We report here the presence in pea (Pisum sativum) mitochondria and nuclei of a Trx isoform (PsTrxo1) that seems to belong to the Trx o group, although it differs from this Trx type by its absence of introns in the genomic sequence. Western-blot analysis with isolated mitochondria and nuclei, immunogold labeling, and green fluorescent protein fusion constructs all indicated that PsTrxo1 is present in both cell compartments. Moreover, the identification by tandem mass spectrometry of the native mitochondrial Trx after gel filtration using the fast-protein liquid chromatography system of highly purified mitochondria and the in vitro uptake assay into isolated mitochondria also corroborated a mitochondrial location for this protein. The recombinant PsTrxo1 protein has been shown to be reduced more effectively by the Saccharomyces cerevisiae mitochondrial Trx reductase Trr2 than by the wheat (Triticum aestivum) cytoplasmic reduced nicotinamide adenine dinucleotide phosphate-dependent Trx reductase. PsTrxo1 was able to activate alternative oxidase, and it was shown to interact with a number of mitochondrial proteins, including peroxiredoxin and enzymes mainly involved in the photorespiratory process. PMID:19363090

  4. Programmed cell death in plants: ultrastructural changes in pea guard cells.

    PubMed

    Bakeeva, L E; Dzyubinskaya, E V; Samuilov, V D

    2005-09-01

    Treatment with cyanide of epidermal peels isolated from pea leaves resulted in destruction of nuclei in the guard cells of stomata, which is visible with a light microscope. The process was accelerated by illumination. Electron microscopy revealed significant CN--induced changes in the ultrastructure of guard cells, which increased with time. Margination of chromatin, which is one of the first signs of apoptosis, was observed in the guard cells even after 1 h incubation of the isolated epidermis with CN-. Subsequent chromatin condensation, swelling of the endoplasmic reticulum with formation of large tanks covered with ribosomes, changes in the structure of dictyosomes, and a slight swelling of mitochondria were observed after 3 h of the epidermis incubation with CN-. After 6 h of incubation with CN-, the bulk volume of the guard cells was filled with vacuoles, the cytoplasm occupied the thin marginal layer, the nucleus was in the center similarly to the control experiment, but it was polylobal, extended in narrow cytoplasmic bands, and, despite the loss of the nuclear envelope integrity, appeared to be a self-dependent structure. In the envelope-free open regions of the nucleus, mitochondria and chloroplasts directly contacted with chromatin. Much like the cell nucleus, chloroplasts lost the integrity of the membrane, but did not swell and retained the stroma and integrity of the thylakoid system. An antioxidant di-tert-butyl-4-hydroxytoluene prevented ultrastructural changes in the cells observed after 6 h of incubation with CN-. Thus, the CN--induced death of the guard cells of stomata occurs through the mechanism of apoptosis.

  5. A novel plasmid pEA68 of Erwinia amylovora and the description of a new family of plasmids.

    PubMed

    Ismail, Emadeldeen; Blom, Jochen; Bultreys, Alain; Ivanović, Milan; Obradović, Aleksa; van Doorn, Joop; Bergsma-Vlami, Maria; Maes, Martine; Willems, Anne; Duffy, Brion; Stockwell, Virginia O; Smits, Theo H M; Puławska, Joanna

    2014-12-01

    Recent genome analysis of Erwinia amylovora, the causal agent of fire blight disease on Rosaceae, has shown that the chromosome is highly conserved among strains and that plasmids are the principal source of genomic diversity. A new circular plasmid, pEA68, was found in E. amylovora strain 692 (LMG 28361), isolated in Poland from Sorbus (mountain ash) with fire blight symptoms. Annotation of the 68,763-bp IncFIIa-type plasmid revealed that it contains 79 predicted CDS, among which two operons (tra, pil) are associated with mobility. The plasmid is maintained stably in E. amylovora and does not possess genes associated with antibiotic resistance or known virulence genes. Curing E. amylovora strain 692 of pEA68 did not influence its virulence in apple shoots nor amylovoran synthesis. Of 488 strains of E. amylovora from seventeen countries, pEA68 was only found in two additional strains from Belgium. Although the spread of pEA68 is currently limited to Europe, pEA68 comprises, together with pEA72 and pEA78 both found in North America, a new plasmid family that spans two continents.

  6. Turgor-responsive gene transcription and RNA levels increase rapidly when pea shoots are wilted. Sequence and expression of three inducible genes.

    PubMed

    Guerrero, F D; Jones, J T; Mullet, J E

    1990-07-01

    Reduction of turgor in pea shoots caused the accumulation of several poly(A) RNAs. cDNA clones derived from three different poly(A) RNAs which accumulate in wilted pea shoots were isolated, sequenced and expression of the corresponding genes examined. Clone 7a encoded a 289 amino acid protein. The C-terminal 180 amino acids of this protein were homologous to soybean nodulin-26. RNA hybridizing to cDNA 7a was abundant in roots, and induced in shoots by dehydration, heat shock and to a small extent by ABA. Hydropathic plots indicate that the protein encoded by cDNA 7a contains six potential membrane spanning domains similar to proteins which form ion channels. Clone 15a encoded a 363 amino acid protein with high homology to cysteine proteases. RNA hybridizing to cDNA 15a was more abundant in roots than shoots of control plants. Dehydration of pea shoots induced cDNA 15a mRNA levels whereas heat shock or ABA treatment did not. Clone 26g encoded a 508 amino acid protein with 30% residue identity to several aldehyde dehydrogenases. RNA hybridizing to cDNA 26g was induced by dehydration of shoots but not roots and heat shock and ABA did not modulate RNA levels. Levels of the three poly(A) RNAs increased 4-6-fold by 4 h after wilting and this increase was not altered by pretreatment of shoots with cycloheximide. When wilted shoots were rehydrated, RNA hybridizing to cDNA 26g declined to pre-stress levels within 2 h. Run-on transcription experiments using nuclei from pea shoots showed that transcription of the genes which encode the three poly(A) RNAs was induced within 30 min following reduction of shoot turgor. One of the genes showed a further increase in transcription by 4 h after dehydration whereas transcription of the other 2 genes declined. These results indicate that plant cells respond to changes in cell turgor by rapidly increasing transcription of several genes. Furthermore, the expression of the turgor-responsive genes varies with respect to the time course of

  7. [Ultrastructure and metabolic activity of pea mitochondria under clinorotation].

    PubMed

    Brykov, V A; Generozova, I P; Shugaev, A G

    2012-01-01

    Experimental data on the mitochondrial ultrastructure and tissue respiration in root apex as well as metabolic activity of the organelles isolated from pea seedling roots after 5-day of clinorotation are presented. It was shown that mitochondrial condensation in the distal elongation zone correlated with an increased rate of oxygen uptake on 7%. We also observed increase in rate of malate oxidation and respiratory control ratio increased simultaneously with a decreased in efficiency of oxidative phosphorylation. Such character of mitochondrial rearrangements in simulated microgravity is assumed to be a consequence of adaptation to these conditions.

  8. Fatty acid biosynthesis in pea root plastids

    SciTech Connect

    Stahl, R.J.; Sparace, S.A. )

    1989-04-01

    Fatty acid biosynthesis from (1-{sup 14}C)acetate was optimized in plastids isolated from primary root tips of 7-day-old germinating pea seeds. Fatty acid synthesis was maximum at approximately 80 nmoles/hr/mg protein in the presence of 200 {mu}M acetate, 0.5 mM each of NADH, NADPH and CoA, 6 mM each of ATP and MgCl{sub 2}, 1 mM each of the MnCl{sub 2} and glycerol-3-phosphate, 15 mM KHCO{sub 3}, and 0.1M Bis-tris-propane, pH 8.0 incubated at 35C. At the standard incubation temperature of 25C, fatty acid synthesis was linear from up to 6 hours with 80 to 100 {mu}g/mL plastid protein. ATP and CoA were absolute requirements, whereas KHCO{sub 3}, divalent cations and reduced nucleotides all improved activity by 80 to 85%. Mg{sup 2+} and NADH were the preferred cation and nucleotide, respectively. Dithiothreitol and detergents were generally inhibitory. The radioactive products of fatty acid biosynthesis were approximately 33% 16:0, 10% 18:0 and 56% 18:1 and generally did not vary with increasing concentrations of each cofactor.

  9. Bean alpha-amylase inhibitors in transgenic peas inhibit development of pea weevil larvae.

    PubMed

    de Sousa-Majer, Maria José; Hardie, Darryl C; Turner, Neil C; Higgins, Thomas J V

    2007-08-01

    This glasshouse study used an improved larval measurement procedure to evaluate the impact of transgenic pea, Pisum sativum L., seeds expressing a-amylase inhibitor (AI)-1 or -2 proteins on pea weevil, Bruchus pisorum L. Seeds of transgenic 'Laura' and 'Greenfeast' peas expressing alpha-(AI)-1 reduced pea weevil survival by 93-98%. Larval mortality occurred at an early instar. Conversely, in nontransgenic cultivars, approximately 98-99% of the pea weevils emerged as adults. By measuring the head capsule size, we determined that larvae died at the first to early third instar in alpha-(AI)-1 transgenic peas, indicating that this inhibitor is highly effective in controlling this insect. By contrast, transgenic Laura and 'Dundale' expressing alpha-(AI)-2 did not affect pea weevil survival, but they did delay larval development. After 77 d of development, the head capsule size indicated that the larvae were still at the third instar stage in transgenic alpha-(AI)-2 peas, whereas adult bruchids had developed in the nontransgenic peas.

  10. Symbiotic activity of pea (Pisum sativum) after application of Nod factors under field conditions.

    PubMed

    Siczek, Anna; Lipiec, Jerzy; Wielbo, Jerzy; Kidaj, Dominika; Szarlip, Paweł

    2014-04-29

    Growth and symbiotic activity of legumes are mediated by Nod factors (LCO, lipo-chitooligosaccharides). To assess the effects of application of Nod factors on symbiotic activity and yield of pea, a two-year field experiment was conducted on a Haplic Luvisol developed from loess. Nod factors were isolated from Rhizobium leguminosarum bv. viciae strain GR09. Pea seeds were treated with the Nod factors (10⁻¹¹ M) or water (control) before planting. Symbiotic activity was evaluated by measurements of nitrogenase activity (acetylene reduction assay), nodule number and mass, and top growth by shoot mass, leaf area, and seed and protein yield. Nod factors generally improved pea yield and nitrogenase activity in the relatively dry growing season 2012, but not in the wet growing season in 2013 due to different weather conditions.

  11. Symbiotic Activity of Pea (Pisum sativum) after Application of Nod Factors under Field Conditions

    PubMed Central

    Siczek, Anna; Lipiec, Jerzy; Wielbo, Jerzy; Kidaj, Dominika; Szarlip, Paweł

    2014-01-01

    Growth and symbiotic activity of legumes are mediated by Nod factors (LCO, lipo-chitooligosaccharides). To assess the effects of application of Nod factors on symbiotic activity and yield of pea, a two-year field experiment was conducted on a Haplic Luvisol developed from loess. Nod factors were isolated from Rhizobium leguminosarum bv. viciae strain GR09. Pea seeds were treated with the Nod factors (10−11 M) or water (control) before planting. Symbiotic activity was evaluated by measurements of nitrogenase activity (acetylene reduction assay), nodule number and mass, and top growth by shoot mass, leaf area, and seed and protein yield. Nod factors generally improved pea yield and nitrogenase activity in the relatively dry growing season 2012, but not in the wet growing season in 2013 due to different weather conditions. PMID:24786094

  12. Evaluation of fluoranthene phytotoxicity in pea plants by Hill reaction and chlorophyll fluorescence.

    PubMed

    Kummerová, Marie; Krulová, Jana; Zezulka, Stepán; Tríska, Jan

    2006-10-01

    The effect of both increased concentrations (0.01 and 1 mg l(-1)) of fluoranthene (FLT) and the duration of exposure (18 and 25 days) on the growth and photosynthetic processes in pea plants (Pisum sativum L., cv. Garde) was investigated. FLT concentration in roots and shoot of pea plants was also determined. The obtained results demonstrated that the higher concentration of FLT (1 mg l(-1)) significantly inhibited the growth of the pea plants after 25 days of the application, also affected the content of photosynthetic pigments (chlorophyll a, b and carotenoids), and the primary photochemical processes of photosynthesis. In chlorophyll fluorescence parameters, the significant increase of F(0) values and the decrease of F(V)/F(M) and Phi(II) values was recorded. The Hill reaction of isolated chloroplasts of pea plants was significantly inhibited after 25 days by presence of FLT (0.01 and 1 mg l(-1)) in nutrient solution, while after 18 days no significant response of Hill reaction activity was recorded. The fluoranthene content in roots and shoot of pea plants increased with increasing FLT concentration in the environment and the substantial accumulation of FLT was observed in the roots.

  13. Isolation of DNA methyltransferase from plants

    SciTech Connect

    Ehrlich, K.; Malbroue, C.

    1987-05-01

    DNA methyltransferases (DMT) were isolated from nuclei of cauliflower, soybean, and pea by extraction with 0.35 M NaCl. Assays were performed on hemimethylated Micrococcus luteus DNA or on M. luteus DNA to test for maintenance or de novo methylase activity, respectively. Fully methylated DNA was used as a substrate to determine background levels of methylation. Based on these tests, yields of maintenance DMT activity in the crude extract from pea hypocotyl, soybean hypocotyl, and cauliflower inflorescence were 2.8, 0.9, and 1.6 units per g wet tissue (one unit equals 1 pmol of methyl from (/sup 3/H)AdoMet incorporated into acid precipitable material per h at 30/sup 0/). Two peaks of DMT activity were detected in the soybean nuclear extract following phosphocellulose chromatography. One eluted at 0.4 M and the other at 0.8 M KCl. With both fractions maintenance activity was approximately 2 times that of the de novo activity. Using gel filtration the DMT eluted at 220,000 Daltons. The optimal pH for activity was between 6.5 and 7.0, and the optimal temperature was 30/sup 0/.

  14. Plant-Parasitic Nematodes and Fungi Associated with Root Rot of Peas on Prince Edward Island

    PubMed Central

    Celetti, M. J.; Johnston, H. W.; Kimpinski, J.; Platt, H. W.

    1990-01-01

    Eight commercial pea fields on Prince Edward Island were sampled in June and July over a 2-year period (1986-87) to determine soil population densities and the incidence of nematodes and fungi associated with root rot of peas. Root lesion nematodes (Pratylenchus spp.) were the dominant endoparasitic nematodes recovered from roots and soil. Low populations of the northern root-knot nematode (Meloidogyne hapla) were also present. Tylenchorhynchus spp. and Paratylenchus spp. were recovered frequently from soil in the root zone, and Helicotylenchus spp. were also frequent, but in low numbers. Fusarium solani was the most common fungal species isolated from the epicotyl and hypocotyl tissues of pea. Fusarium oxysporum was also isolated frequently, and both Fusarium species were found in soil from all fields. Rhizoctonia solani and Verticillium albo-atrum were common in hypocotyl tissue, but V. dahliae was isolated infrequently. Root rot was rated as severe in all fields and was positively and significantly correlated (P ≤ 0.05) with densities of Tylenchorhynchus spp. in soil and with incidence of F. solani in pea tissue. The incidence of F. solani root infections was positively and significantly correlated with densities in soil of Tylenchorhynchus spp. (P ≤ 0.01), Helicotylenchus spp. (P ≤ 0.01), and Paratylenchus spp. (P ≤ 0.05). PMID:19287779

  15. Overexpression of the ped/pea-15 gene causes diabetes by impairing glucose-stimulated insulin secretion in addition to insulin action.

    PubMed

    Vigliotta, Giovanni; Miele, Claudia; Santopietro, Stefania; Portella, Giuseppe; Perfetti, Anna; Maitan, Maria Alessandra; Cassese, Angela; Oriente, Francesco; Trencia, Alessandra; Fiory, Francesca; Romano, Chiara; Tiveron, Cecilia; Tatangelo, Laura; Troncone, Giancarlo; Formisano, Pietro; Beguinot, Francesco

    2004-06-01

    Overexpression of the ped/pea-15 gene is a common feature of type 2 diabetes. In the present work, we show that transgenic mice ubiquitously overexpressing ped/pea-15 exhibited mildly elevated random-fed blood glucose levels and decreased glucose tolerance. Treatment with a 60% fat diet led ped/pea-15 transgenic mice to develop diabetes. Consistent with insulin resistance in these mice, insulin administration reduced glucose levels by only 35% after 45 min, compared to 70% in control mice. In vivo, insulin-stimulated glucose uptake was decreased by almost 50% in fat and muscle tissues of the ped/pea-15 transgenic mice, accompanied by protein kinase Calpha activation and block of insulin induction of protein kinase Czeta. These changes persisted in isolated adipocytes from the transgenic mice and were rescued by the protein kinase C inhibitor bisindolylmaleimide. In addition to insulin resistance, ped/pea-15 transgenic mice showed a 70% reduction in insulin response to glucose loading. Stable overexpression of ped/pea-15 in the glucose-responsive MIN6 beta-cell line also caused protein kinase Calpha activation and a marked decline in glucose-stimulated insulin secretion. Antisense block of endogenous ped/pea-15 increased glucose sensitivity by 2.5-fold in these cells. Thus, in vivo, overexpression of ped/pea-15 may lead to diabetes by impairing insulin secretion in addition to insulin action.

  16. Finding linked markers to En for efficient selection of pea enation Mosaic Virus resistance in pea

    USDA-ARS?s Scientific Manuscript database

    Pea enation mosaic virus (PEMV) causes an important disease of cool season food legumes, resulting in significant yield loss worldwide. The present investigation was carried out to study the inheritance and identify the molecular markers linked with the PEMV resistance gene (En) in field pea (Pisum ...

  17. Peas in a Pod: Environment and Ionization in Green Pea Galaxies

    NASA Astrophysics Data System (ADS)

    Kurtz, Heather; Jaskot, Anne; Drew, Patrick; Pare, Dylan; Griffin, Jon; Petersen, Michael

    2016-01-01

    The Green Peas are extreme, highly ionized, starburst galaxies with strong [OIII] 5007 emission. Using the Sloan Digital Sky Survey, we present statistics on the environment of Green Peas and investigate its effects on their ionized gas properties. Although most dwarf starburst galaxies are in low-density environments, we identify a sample of Green Peas in dense environments. Emission line observations with the WIYN 0.9-meter telescope at Kitt Peak reveal that one cluster Green Pea is more highly ionized in the direction of the cluster center. Ram pressure stripping likely generates this ionization gradient. We explore the role of the environment in enhancing star formation rates and ionization, and we compare the nebular properties of Green Peas in high-density environments to those in low-density environments.

  18. Arthropod diversity in peas with normal or reduced waxy bloom

    PubMed Central

    Chang, Gary C.; Rutledge, Claire E.; Biggam, Russell C.; Eigenbrode, Sanford D.

    2004-01-01

    Crop traits can alter economically important interactions between plants, pests, and biological control agents. For example, a reduced waxy bloom on the surface of pea plants alters interactions between pea aphids and their natural enemies. In this study, we assess whether the effect of wax reduction extends beyond the 2 or 3 arthropod species closely associated with the plants and into the structure of the broader arthropod community of over 200 taxa at our site. We sampled arthropods on lines of peas with normal and reduced wax in Latah Co., Idaho using pitfall traps within randomly assigned pairs of 5 × 5 meter plots. During the 1998 and 1999 growing seasons, we collected 12,113 individual arthropods from 221 unambiguously identified morphospecies. The number of individuals collected from each morphospecies responded idiosyncratically to the reduced wax peas. To test whether arthropod community structure differed between the collections from plots having peas with normal or reduced wax, we performed a randomization test. The collection from peas with reduced wax had higher species evenness and thus higher community diversity despite having lower species richness. Our results demonstrate the potential of a single plant trait, epicuticular wax, to affect a community of arthropods. Two pests of peas had opposite responses to peas with reduced wax. The number of pea aphids collected was greater from peas with normal wax peas than those with reduced wax. In contrast, the number of pea leaf weevils collected was greater from peas with reduced wax. PMID:15861234

  19. Stamina pistilloida, the Pea ortholog of Fim and UFO, is required for normal development of flowers, inflorescences, and leaves.

    PubMed

    Taylor, S; Hofer, J; Murfet, I

    2001-01-01

    Isolation and characterization of two severe alleles at the Stamina pistilloida (Stp) locus reveals that Stp is involved in a wide range of developmental processes in the garden pea. The most severe allele, stp-4, results in flowers consisting almost entirely of sepals and carpels. Production of ectopic secondary flowers in stp-4 plants suggests that Stp is involved in specifying floral meristem identity in pea. The stp mutations also reduce the complexity of the compound pea leaf, and primary inflorescences often terminate prematurely in an aberrant sepaloid flower. In addition, stp mutants were shorter than their wild-type siblings due to a reduction in cell number in their internodes. Fewer cells were also found in the epidermis of the leaf rachis of stp mutants. Examination of the effects of stp-4 in double mutant combinations with af, tl, det, and veg2-2-mutations known to influence leaf, inflorescence, and flower development in pea-suggests that Stp function is independent of these genes. A synergistic interaction between weak mutant alleles at Stp and Uni indicated that these two genes act together, possibly to regulate primordial growth. Molecular analysis revealed that Stp is the pea homolog of the Antirrhinum gene Fimbriata (Fim) and of UNUSUAL FLORAL ORGANS (UFO) from Arabidopsis. Differences between Fim/UFO and Stp mutant phenotypes and expression patterns suggest that expansion of Stp activity into the leaf was an important step during evolution of the compound leaf in the garden pea.

  20. Emulsifying and foaming properties of commercial yellow pea (Pisum sativum L.) seed flours.

    PubMed

    Aluko, Rotimi E; Mofolasayo, Olawunmi A; Watts, Beverley M

    2009-10-28

    Commercial yellow pea seed flours prepared by a patented wet-milling process and pea protein isolate (PPI) were analyzed for emulsifying and foaming properties at pH 3.0, 5.0, and 7.0 and compared to soybean protein isolate (SPI). PPI and SPI formed emulsions with significantly smaller (p < 0.05) oil droplet sizes, 16-30 and 23-54 microm, respectively, than flours that primarily contained fiber such as Centara III and IV, or those that consisted mainly of starch: Centu-tex, Uptake 80 and Accu-gel. PPI was a better emulsifier than SPI at pH 7.0, and a better foaming agent at pH 3.0 and pH 7.0, although foaming capacity varied with sample concentration. Centu-tex and Uptake 80 have exactly the same chemical composition, but the latter has a much smaller flour particle size range, and had significantly smaller (p < 0.05) emulsion oil droplets. Incorporation of pea starch into SPI emulsions produced a synergistic effect that led to significant increases (p < 0.05) in emulsification capacity (reduced emulsion oil droplet size) when compared to SPI or starch alone. These results showed that PPI had generally significantly higher (p < 0.05) emulsion and foam forming properties than SPI, and that pea starch could be used to improve the quality of SPI-stabilized food emulsions.

  1. Light activation of fructose bisphosphatase in photosynthetically competent pea chloroplasts.

    PubMed

    Charles, S A; Halliwell, B

    1981-11-15

    The fructose bisphosphatase (EC 3.1.3.11) activity of type A chloroplasts isolated from young (9-day-old) pea (Pisum sativum var. Progress no. 9) plants, assayed at physiological pH, substrate and Mg2+ concentrations, increased rapidly on illumination. The enzyme activity detected was more than sufficient to account for observed rates of Co2 fixation both during the induction period and during steady-state CO2 fixation, whether or not dihydroxyacetone phosphate had been added to the preparation. Omission of catalase from the suspension medium had no effect. On switching off the light, CO2 fixation by the chloroplasts ceased at once, yet fructose bisphosphatase activity decreased much more slowly. Changes in enzyme activity were much less marked if assays were conducted at 3 mM substrate and 10 mM-Mg2+. Chloroplasts from older (13--20-day-old) peas only fixed CO2 rapidly if catalase was present in the assay medium. The fructose bisphosphatase activity detected under physiological assay conditions was again more than sufficient to account for observed rates of Co2 fixation. In the presence of added dihydroxyacetone phosphate, however, the rate of Co2 fixation appeared to be determined by the rate of light activation of fructose bisphosphatase. In general, the rates of Co2 fixation and enzyme activation, and the final enzyme activity achieved, decreased markedly with increasing age of the plants. The role of light activation of fructose bisphosphatase as a means of controlling the rate of CO2 fixation in pea chloroplasts is discussed.

  2. Light activation of fructose bisphosphatase in photosynthetically competent pea chloroplasts.

    PubMed Central

    Charles, S A; Halliwell, B

    1981-01-01

    The fructose bisphosphatase (EC 3.1.3.11) activity of type A chloroplasts isolated from young (9-day-old) pea (Pisum sativum var. Progress no. 9) plants, assayed at physiological pH, substrate and Mg2+ concentrations, increased rapidly on illumination. The enzyme activity detected was more than sufficient to account for observed rates of Co2 fixation both during the induction period and during steady-state CO2 fixation, whether or not dihydroxyacetone phosphate had been added to the preparation. Omission of catalase from the suspension medium had no effect. On switching off the light, CO2 fixation by the chloroplasts ceased at once, yet fructose bisphosphatase activity decreased much more slowly. Changes in enzyme activity were much less marked if assays were conducted at 3 mM substrate and 10 mM-Mg2+. Chloroplasts from older (13--20-day-old) peas only fixed CO2 rapidly if catalase was present in the assay medium. The fructose bisphosphatase activity detected under physiological assay conditions was again more than sufficient to account for observed rates of Co2 fixation. In the presence of added dihydroxyacetone phosphate, however, the rate of Co2 fixation appeared to be determined by the rate of light activation of fructose bisphosphatase. In general, the rates of Co2 fixation and enzyme activation, and the final enzyme activity achieved, decreased markedly with increasing age of the plants. The role of light activation of fructose bisphosphatase as a means of controlling the rate of CO2 fixation in pea chloroplasts is discussed. PMID:6280684

  3. Immunofluorescence detection of pea protein in meat products.

    PubMed

    Petrášová, Michaela; Pospiech, Matej; Tremlová, Bohuslava; Javůrková, Zdeňka

    2016-08-01

    In this study we developed an immunofluorescence method to detect pea protein in meat products. Pea protein has a high nutritional value but in sensitive individuals it may be responsible for causing allergic reactions. We produced model meat products with various additions of pea protein and flour; the detection limit (LOD) of the method for pea flour was 0.5% addition, and for pea protein it was 0.001% addition. The repeatabilities and reproducibilities for samples both positive and negative for pea protein were all 100%. In a blind test with model products and commercial samples, there was no statistically significant difference (p > 0.05) between the declared concentrations of pea protein and flour and the immunofluorescence method results. Sensitivity was 1.06 and specificity was 1.00. These results show that the immunofluorescence method is suitable for the detection of pea protein in meat products.

  4. NORTHWEST OBLIQUE AERIAL VIEW OF FORT DELAWARE AND PEA PATCH ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    NORTHWEST OBLIQUE AERIAL VIEW OF FORT DELAWARE AND PEA PATCH ISLAND. REMAINS OF SEA WALL VISIBLE IN FOREGROUND AND RIGHT OF IMAGE - Fort Delaware, Pea Patch Island, Delaware City, New Castle County, DE

  5. 1. NORTHWEST OBLIQUE AERIAL VIEW OF FORT DELAWARE AND PEA ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    1. NORTHWEST OBLIQUE AERIAL VIEW OF FORT DELAWARE AND PEA PATCH ISLAND. REMAINS OF SEA WALL VISIBLE IN FOREGROUND AND RIGHT OF IMAGE. - Fort Delaware, Sea Wall, Pea Patch Island, Delaware City, New Castle County, DE

  6. PED/PEA-15 Inhibits Hydrogen Peroxide-Induced Apoptosis in Ins-1E Pancreatic Beta-Cells via PLD-1

    PubMed Central

    Raciti, Gregory Alexander; Zatterale, Federica; Nigro, Cecilia; Mirra, Paola; Falco, Roberta; Ulianich, Luca; Di Jeso, Bruno; Formisano, Pietro; Miele, Claudia; Beguinot, Francesco

    2014-01-01

    The small scaffold protein PED/PEA-15 is involved in several different physiologic and pathologic processes, such as cell proliferation and survival, diabetes and cancer. PED/PEA-15 exerts an anti-apoptotic function due to its ability to interfere with both extrinsic and intrinsic apoptotic pathways in different cell types. Recent evidence shows that mice overexpressing PED/PEA-15 present larger pancreatic islets and increased beta-cells mass. In the present work we investigated PED/PEA-15 role in hydrogen peroxide-induced apoptosis in Ins-1E beta-cells. In pancreatic islets isolated from TgPED/PEA-15 mice hydrogen peroxide-induced DNA fragmentation was lower compared to WT islets. TUNEL analysis showed that PED/PEA-15 overexpression increases the viability of Ins-1E beta-cells and enhances their resistance to apoptosis induced by hydrogen peroxide exposure. The activity of caspase-3 and the cleavage of PARP-1 were markedly reduced in Ins-1E cells overexpressing PED/PEA-15 (Ins-1EPED/PEA-15). In parallel, we observed a decrease of the mRNA levels of pro-apoptotic genes Bcl-xS and Bad. In contrast, the expression of the anti-apoptotic gene Bcl-xL was enhanced. Accordingly, DNA fragmentation was higher in control cells compared to Ins-1EPED/PEA-15 cells. Interestingly, the preincubation with propranolol, an inhibitor of the pathway of PLD-1, a known interactor of PED/PEA-15, responsible for its deleterious effects on glucose tolerance, abolishes the antiapoptotic effects of PED/PEA-15 overexpression in Ins-1E beta-cells. The same results have been obtained by inhibiting PED/PEA-15 interaction with PLD-1 in Ins-1EPED/PEA-15. These results show that PED/PEA-15 overexpression is sufficient to block hydrogen peroxide-induced apoptosis in Ins-1E cells through a PLD-1 mediated mechanism. PMID:25489735

  7. Potential alternative hosts for a powdery mildew on pea

    USDA-ARS?s Scientific Manuscript database

    Powdery mildew of pea (Pisum sativum) is an important disease in the field and in the greenhouse. The most widely documented powdery mildew on pea is Erysiphe pisi, but E. trifolii and E. baeumleri have also been reported. From greenhouse-grown peas, we obtained powdery mildew samples with rDNA ITS ...

  8. Pea Disease Diagnostic Series- Rhizoctonia seed, seedling and root rot

    USDA-ARS?s Scientific Manuscript database

    Pea disease diagnostic cards that growers can carry with them into the field that are water resistant and durable which can be used to identify the signs and symptoms of major pea pathogens were developed. Color photographs of major fungal, bacterial, and viral pathogens on peas and a brief descript...

  9. The Pea Gene LH Encodes ent-Kaurene Oxidase1

    PubMed Central

    Davidson, Sandra E.; Smith, Jennifer J.; Helliwell, Chris A.; Poole, Andrew T.; Reid, James B.

    2004-01-01

    The pea (Pisum sativum) homolog, PsKO1, of the Arabidopsis GA3 gene was isolated. It codes for a cytochrome P450 from the CYP701A subfamily and has ent-kaurene oxidase (KO) activity, catalyzing the three step oxidation of ent-kaurene to ent-kaurenoic acid in the gibberellin (GA) biosynthetic pathway when expressed in yeast (Saccharomyces cerevisiae). PsKO1 is encoded by the LH gene because in three independent mutant alleles, lh-1, lh-2, and lh-3, PsKO1 has altered sequence, and the lh-1 allele, when expressed in yeast, failed to metabolize ent-kaurene. The lh mutants of pea are GA deficient and have reduced internode elongation and root growth. One mutant (lh-2) also causes a large increase in seed abortion. PsKO1 (LH) is expressed in all tissues examined, including stems, roots, and seeds, and appears to be a single-copy gene. Differences in sensitivity to the GA synthesis inhibitor, paclobutrazol, between the mutants appear to result from the distinct nature of the genetic lesions. These differences may also explain the tissue-specific differences between the mutants. PMID:14988475

  10. Fluidity of pea root plasma membranes under altered gravity

    NASA Astrophysics Data System (ADS)

    Klymchuk, D. O.; Baranenko, V. V.; Vorobyova, T. V.; Dubovoy, V. D.

    This investigation aims to determine whether clinorotation 2 rev min of pea Pisum sativum L seedlings induces the alterations in the physical-chemical properties of cellular membranes including the plasma membrane fluidity The last is an important regulator of functional activity of membrane enzymes The plasma membranes were isolated by aqueous two-phase partitioning from roots of 6-day old pea seedlings The membrane fluidity was examined by fluorescence spectroscopy using pyrene probe The plasma membrane vesicles with known protein concentration were added to the incubation buffer to a final concentration of 50 mu g of protein per ml A small amount by 1 mu l of pyrene solution in 2-propanol was added to the incubation mixture to a final probe concentration 5 mu M at constant mixing Fluorescence spectra were measured using a Perkin-Elmer LS-50 spectrofluorometer Perkin-Elmer England Pyrene was excited at 337 nm and fluorescence intensity of monomers I M and excimers I E were measured at 393 and 470 nm respectively The I E I M ratios were 0 081 pm 0 003 and 0 072 pm 0 004 in preparations obtained from clinorotated and the control seedlings respectively This fact indicates that rotation on the clinostat increases the membrane fluidity Compared with controls clinorotated seedlings have also showed a reduced growth and a higher level of total unsaturated fatty acids determined by gas chromatography The factors that influence on the fluidity of membrane lipids in bilayer appear to be the

  11. The pea gene LH encodes ent-kaurene oxidase.

    PubMed

    Davidson, Sandra E; Smith, Jennifer J; Helliwell, Chris A; Poole, Andrew T; Reid, James B

    2004-03-01

    The pea (Pisum sativum) homolog, PsKO1, of the Arabidopsis GA3 gene was isolated. It codes for a cytochrome P450 from the CYP701A subfamily and has ent-kaurene oxidase (KO) activity, catalyzing the three step oxidation of ent-kaurene to ent-kaurenoic acid in the gibberellin (GA) biosynthetic pathway when expressed in yeast (Saccharomyces cerevisiae). PsKO1 is encoded by the LH gene because in three independent mutant alleles, lh-1, lh-2, and lh-3, PsKO1 has altered sequence, and the lh-1 allele, when expressed in yeast, failed to metabolize ent-kaurene. The lh mutants of pea are GA deficient and have reduced internode elongation and root growth. One mutant (lh-2) also causes a large increase in seed abortion. PsKO1 (LH) is expressed in all tissues examined, including stems, roots, and seeds, and appears to be a single-copy gene. Differences in sensitivity to the GA synthesis inhibitor, paclobutrazol, between the mutants appear to result from the distinct nature of the genetic lesions. These differences may also explain the tissue-specific differences between the mutants.

  12. Genetic variation in pea seed globulin composition.

    PubMed

    Tzitzikas, Emmanouil N; Vincken, Jean-Paul; de Groot, Jolan; Gruppen, Harry; Visser, Richard G F

    2006-01-25

    A quantitative characterization of seeds from 59 pea (Pisum sativum L.) lines and relative taxa with various external characteristics and wide geographical origin was performed to explore the genetic variation of pea concerning its starch and protein contents and globulin composition. Pea lines, which produce round, wrinkled, flat, and round-dimpled seeds, have starch as the major reserve, with an average content of 46%. Protein content varied from 13.7 to 30.7% of the seed dry matter, with an overall average of 22.3%. Densitometric quantification of the individual globulins (legumin, vicilin, convicilin, and globulin-related proteins) based on SDS-PAGE gels showed no lines lacking any particular globulin. Among the lines tested, variation was shown in both their total globulins content and their globulin composition. The total globulin content ranged from 49.2 to 81.8% of the total pea protein extract (TPPE). Legumin content varied between 5.9 and 24.5% of the TPPE. Vicilin was the most abundant protein of pea, and its content varied between 26.3 and 52.0% of the TPPE. Both processed and nonprocessed vicilins occurred. The processed vicilin was the predominant one, with values between 17.8 and 40.8%, whereas the nonprocessed ones constituted between 3.1 and 13.5% of the TPPE. Convicilin was the least abundant globulin, and its content ranged from 3.9 to 8.3%. Finally, the globulin-related proteins were present in amounts ranging from 2.8 to 17.3%. They were less abundant in comparison with legumin and vicilin, but they showed the largest relative variation of the four globulin classes. Correlations between the different external characteristics and globulin composition were determined. Comparison with soybean showed that pea lines show more variety in the abundance of globulin proteins, enabling a wider range of food application.

  13. Tamoxifen and related compounds protect against lipid peroxidation in isolated nuclei: relevance to the potential anticarcinogenic benefits of breast cancer prevention and therapy with tamoxifen?

    PubMed

    Wiseman, H; Halliwell, B

    1994-11-01

    Tamoxifen, 4-hydroxytamoxifen, nafoxidine, 17 beta-oestradiol and ICI 164,384 were all found to protect rat liver nuclei against Fe(III)-ascorbate dependent lipid peroxidation. The order of effectiveness of these compounds was 4-hydroxytamoxifen > 17 beta-oestradiol > nafoxidine > tamoxifen > ICI 164,384. This protection by tamoxifen against the formation of the genotoxic reactive-intermediates and products of lipid peroxidation in the nuclear membrane could be important in the prevention of nuclear DNA damage and thus carcinogenesis. This possible anticarcinogenic benefit of tamoxifen treatment could be important in long-term therapy with tamoxifen (and future derivatives) and in its proposed use in the prevention of breast cancer.

  14. Evidence that QTL may be involved in a tolerance response to Pea enation mosaic virus in pea (Pisum sativum L.)

    USDA-ARS?s Scientific Manuscript database

    Pea enation mosaic virus (PEMV) is a serious disease of fresh market and dry pea in the Pacific Northwest region of the U.S. The En gene confers resistance to PEMV in pea, however a limited number of available cultivars contain the gene, and sources of tolerance have not been reported. In 2007, ad...

  15. Effect of Ca2+ on programmed death of guard and epidermal cells of pea leaves.

    PubMed

    Kiselevsky, D B; Kuznetsova, Yu E; Vasil'ev, L A; Lobysheva, N V; Zinovkin, R A; Nesov, A V; Shestak, A A; Samuilov, V D

    2010-05-01

    The effect of Ca2+ on programmed death of guard cells (GC) and epidermal cells (EC) determined from destruction of the cell nucleus was investigated in epidermis of pea leaves. Ca2+ at concentrations of 1-100 microM increased and at a concentration of 1 mM prevented the CN(-)-induced destruction of the nucleus in GC, disrupting the permeability barrier of GC plasma membrane for propidium iodide (PI). Ca2+ at concentrations of 0.1-1 mM enhanced drastically the number of EC nuclei stained by PI in epidermis treated with chitosan, an inducer of programmed cell death. The internucleosomal DNA fragmentation caused by CN(-) was suppressed by 2 mM Ca2+ on 6 h incubation, but fragmentation was stimulated on more prolonged treatment (16 h). Presumably, the disruption of the permeability barrier of plasma membrane for PI is not a sign of necrosis in plant cells. Quinacrine and diphenylene iodonium at 50 microM concentration prevented GC death induced by CN(-) or CN(-) + 0.1 mM Ca2+ but had no influence on respiration and photosynthetic O2 evolution in pea leaf slices. The generation of reactive oxygen species determined from 2',7'-dichlorofluorescein fluorescence was promoted by Ca2+ in epidermal peels from pea leaves.

  16. Effect of Xyloglucan Oligosaccharides on Growth, Viscoelastic Properties, and Long-Term Extension of Pea Shoots.

    PubMed Central

    Cutillas-Iturralde, A.; Lorences, E. P.

    1997-01-01

    The growth-promoting effect of xyloglucan-derived oligosaccharides was investigated using a bioassay with entire pea (Pisum sativum L., var Alaska) shoots. After a 24-h incubation period at 25[deg]C, xyloglucan oligosaccharide (XGO) solutions with concentrations of 10-6 M notably increased the growth rate of pea shoots, whereas the same oligosaccharides at 10-7 M were less effective. To investigate the possible correlation between growth rate changes in the XGO-treated shoots and changes in the wall mechanical properties of their growing regions (third internodes), we used a short-term creep assay. The promotion of elongation by XGOs was reflected in an enhancement of the viscoelasticity of the growing regions of the shoots. To show whether this effect on wall viscoelastic properties was the cause or a consequence of their growth promotion, we tested the effect of XGOs on the long-term extension of isolated cell walls. We characterized an acid-induced extension in isolated cell walls from pea shoots that was not inhibited by preincubation in neutral buffers. Exogenously added XGOs did not alter the pattern of pea segment extension at any pH tested, indicating that XGOs have no direct effect on cell wall viscoelasticity. Finally, preincubation of pea segments in neutral buffers with XGOs enhanced their capacity to extend under acidic conditions. This finding suggests that XGOs at a neutral pH can act via transglycosylation, weakening the wall matrix and making the wall more responsive to other mechanisms of acid-induced extension as an expansin-mediated extension. PMID:12223593

  17. Registration of Pea Germplasm Partially Resistant to Aphanomyces Root Rot for Breeding Fresh or Freezer Pea and Dry Pea Types

    USDA-ARS?s Scientific Manuscript database

    Seven F8 derived breeding lines, 846-07, 847-08, 847-22, 847-45, 847-50, 847-53 and 847-68, of green pea (Pisum sativum, L.) were selected from a recombinant inbred line population that was developed by the USDA ARS in 2002. These lines are unique as they combine high levels of tolerance to Aphanom...

  18. Clarification on Host Range of Didymella pinodes the Causal Agent of Pea Ascochyta Blight

    PubMed Central

    Barilli, Eleonora; Cobos, Maria José; Rubiales, Diego

    2016-01-01

    Didymella pinodes is the principal causal agent of ascochyta blight, one of the most important fungal diseases of pea (Pisum sativum) worldwide. Understanding its host specificity has crucial implications in epidemiology and management; however, this has not been clearly delineated yet. In this study we attempt to clarify the host range of D. pinodes and to compare it with that of other close Didymella spp. D. pinodes was very virulent on pea accessions, although differences in virulence were identified among isolates. On the contrary, studied isolates of D. fabae, D. rabiei, and D. lentil showed a reduced ability to infect pea not causing macroscopically visible symptoms on any of the pea accessions tested. D. pinodes isolates were also infective to some extend on almost all species tested including species such as Hedysarum coronarium, Lathyrus sativus, Lupinus albus, Medicago spp., Trifolium spp., Trigonella foenum-graecum, and Vicia articulata which were not mentioned before as hosts of D. pinodes. On the contrary, D. lentil and D. rabiei were more specific, infecting only lentil and chickpea, respectively. D. fabae was intermediate, infecting mainly faba bean, but also slightly other species such as Glycine max, Phaseolus vulgaris, Trifolium spp., Vicia sativa, and V. articulata. DNA sequence analysis of the nuclear ribosomal internal transcribed spacer region (ITS) was performed to confirm identity of the isolates studies and to determine phylogenetic relationship among the Didymella species, revealing the presence of two clearly distinct clades. Clade one was represented by two supported subclusters including D. fabae isolates as well as D. rabiei with D. lentil isolates. Clade two was the largest and included all the D. pinodes isolates as well as Phoma medicaginis var. pinodella. Genetic distance between D. pinodes and the other Didymella spp. isolates was not correlated with overall differences in pathogenicity. Based on evidences presented here, D

  19. Review of the health benefits of peas (Pisum sativum L.).

    PubMed

    Dahl, Wendy J; Foster, Lauren M; Tyler, Robert T

    2012-08-01

    Pulses, including peas, have long been important components of the human diet due to their content of starch, protein and other nutrients. More recently, the health benefits other than nutrition associated with pulse consumption have attracted much interest. The focus of the present review paper is the demonstrated and potential health benefits associated with the consumption of peas, Pisum sativum L., specifically green and yellow cotyledon dry peas, also known as smooth peas or field peas. These health benefits derive mainly from the concentration and properties of starch, protein, fibre, vitamins, minerals and phytochemicals in peas. Fibre from the seed coat and the cell walls of the cotyledon contributes to gastrointestinal function and health, and reduces the digestibility of starch in peas. The intermediate amylose content of pea starch also contributes to its lower glycaemic index and reduced starch digestibility. Pea protein, when hydrolysed, may yield peptides with bioactivities, including angiotensin I-converting enzyme inhibitor activity and antioxidant activity. The vitamin and mineral contents of peas may play important roles in the prevention of deficiency-related diseases, specifically those related to deficiencies of Se or folate. Peas contain a variety of phytochemicals once thought of only as antinutritive factors. These include polyphenolics, in coloured seed coat types in particular, which may have antioxidant and anticarcinogenic activity, saponins which may exhibit hypocholesterolaemic and anticarcinogenic activity, and galactose oligosaccharides which may exert beneficial prebiotic effects in the large intestine.

  20. Analysis of the state of posttranslational calmodulin methylation in developing pea plants. [Pisum sativum

    SciTech Connect

    Oh, Sukheung; Roberts, D.M. )

    1990-07-01

    A specific calmodulin-N-methyltransferase was used in a radiometric assay to analyze the degree of methylation of lysine-115 in pea (Pisum sativum) plants. Calmodulin was isolated from dissected segments of developing roots of young etiolated and green pea plants and was tested for its ability to be methylated by incubation with the calmodulin methyltransferase in the presence of ({sup 3}H)methyl-S-adenosylmethionine. By this approach, the presence of unmethylated calmodulins were demonstrated in pea tissues, and the levels of methylation varied depending on the developmental state of the tissue tested. Calmodulin methylation levels were lower in apical root segments of both etiolated and green plants, and in the young lateral roots compared with the mature, differentiated root tissues. The incorporation of methyl groups into these calmodulin samples appears to be specific for position 115 since site-directed mutants of calmodulin with substitutions at this position competitively inhibited methyl group incorporation. The present findings, combined with previous data showing differences in the ability of methylated and unmethylated calmodulins to activate pea NAD kinase raise the possibility that posttranslational methylation of calmodulin could be another mechanism for regulating calmodulin activity.

  1. CEI-PEA Alert, Summer 2006

    ERIC Educational Resources Information Center

    Center for Educational Innovation - Public Education Association, 2006

    2006-01-01

    The "CEI-PEA Alert" is an advocacy newsletter that deals with topics of interest to all concerned with the New York City public schools. This issue includes: (1) Practical Skills & High Academic Standards: Career Technical Education; (2) Parents: Help Your Children Gain "Soft Skills" for the Workforce; (3) Culinary Arts…

  2. Iron bioavailability in low phytate pea

    USDA-ARS?s Scientific Manuscript database

    Field pea (Pisum sativum L.) seeds have high nutritional value but also contain potential anti-nutritional factors, such as phytate and polyphenols. Phytate can store up to 80% of the phosphorus in seeds. In the seed and during digestion it can complex minerals such as iron and zinc and make them un...

  3. Actin and Myosin in Pea Tendrils 1

    PubMed Central

    Ma, Yong-Ze; Yen, Lung-Fei

    1989-01-01

    We demonstrate here the presence of actin and myosin in pea (Pisum sativum L.) tendrils. The molecular weight of tendril actin is 43,000, the same as rabbit skeletal muscle actin. The native molecular weight of tendril myosin is about 440,000. Tendril myosin is composed of two heavy chains of molecular weight approximately 165,000 and four (two pairs) light chains of 17,000 and 15,000. At high ionic strength, the ATPase activity of pea tendril myosin is activated by K+-EDTA and Ca2+ and is inhibited by Mg2+. At low ionic strength, the Mg2+-ATPase activity of pea tendril myosin is activated by rabbit skeletal muscle F-actin. Superprecipitation occurred after incubation at room temperature when ATP was added to the crude actomyosin extract. It is suggested that the interaction of actin and myosin may play a role in the coiling movement of pea tendril. Images Figure 1 Figure 3 Figure 4 PMID:16666586

  4. CEI-PEA Alert, Fall 2005

    ERIC Educational Resources Information Center

    Center for Educational Innovation - Public Education Association, 2005

    2005-01-01

    The "CEI-PEA Alert" is an advocacy newsletter that deals with topics of interest to all concerned with the New York City public schools. This issue includes: (1) Chancellor Joel I. Klein Announces New Accountability System for NYC Schools; (2) Students Achieve Record-High Scores!; (3) Use Data to Help Your Child Improve Performance; (4)…

  5. Pea disease diagnostic series - White mold

    USDA-ARS?s Scientific Manuscript database

    White mold is a serious disease of pea worldwide, and it is caused by the fungus Sclerotinia sclerotiorum. Water soaked lesions and white mycelial growth may occur on leaves, stems and pods, and are characteristics of the disease. The pathogen may form black fruiting bodies called sclerotia on infec...

  6. 21 CFR 158.170 - Frozen peas.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... than 15 percent by count of the peas may sink in a solution containing 16 percent salt by weight... determination. (i) Extracting solutions: (a) One hundred parts of ethanol denatured with five parts of methanol volume to volume (formula 3A denatured alcohol), or (b) A mixture of 95 parts of formula 3A...

  7. 21 CFR 158.170 - Frozen peas.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... percent by weight, or more than 12 sq cm (2 sq in) in area, extraneous vegetable material, i.e., vine or.... With cloth wipe surplus water from lower screen surface. Weigh 250 g. of peas into high-speed blender.... From this weight deduct weight of dish, cover, and paper. Calculate percent by weight of alcohol...

  8. 21 CFR 155.170 - Canned peas.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... hydrogen ion concentration. (xiii) Seasonings and garnishes: (a) Pieces of green or red peppers or mixtures... designation “green.” The term “early,” “June,” or “early June” shall precede or follow the name in the case of... characteristics. Where the peas are of sweet green wrinkled varieties or hybrids having similar characteristics...

  9. 21 CFR 155.170 - Canned peas.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    .... (a) Identity—(1) Definition. Canned peas is the food prepared from fresh or frozen succulent seeds of... drained weight of the finished food. (b) Lemon juice or concentrated lemon juice. (c) Mint leaves. (d... following shall be included as part of the name or in close proximity to the name of the food: (a)...

  10. 21 CFR 155.170 - Canned peas.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    .... (a) Identity—(1) Definition. Canned peas is the food prepared from fresh or frozen succulent seeds of... drained weight of the finished food. (b) Lemon juice or concentrated lemon juice. (c) Mint leaves. (d... following shall be included as part of the name or in close proximity to the name of the food: (a)...

  11. Pea dehydrins: identification, characterisation and expression.

    PubMed

    Roberton, M; Chandler, P M

    1992-09-01

    An antiserum raised against dehydrin from maize (Zea mays) recognised several polypeptides in extracts of pea (Pisum sativum) cotyledons. A cDNA expression library was prepared from mRNA of developing cotyledons, screened with the antiserum and positive clones were purified and characterised. The nucleotide sequence of one such clone, pPsB12, contained an open reading frame which would encode a polypeptide with regions of significant amino acid sequence similarity to dehydrins from other plant species. The deduced amino acid sequence of the pea dehydrin encoded by B12 is 197 amino acids in length, has a high glycine content (25.9%), lacks tryptophan and is highly hydrophilic. The polypeptide has an estimated molecular mass of 20.4 kDa and pI = 6.4. An in vitro synthesised product from the clone comigrates with one of the in vivo proteins recognised by the antiserum. A comparison of the pea dehydrin sequence with sequences from other species revealed conserved amino acid regions: an N-terminal DEYGNP and a lysine-rich block (KIKEKLPG), both of which are present in two copies. Unexpectedly, pea dehydrin lacks a stretch of serine residues which is conserved in other dehydrins. B12 mRNA and dehydrin proteins accumulated in dehydration-stressed seedlings, associated with elevated levels of endogenous abscisic acid (ABA). Applied ABA induced expression of dehydrins in unstressed seedlings. Dehydrin expression was rapidly reversed when seedlings were removed from the stress or from treatment with ABA and placed in water. During pea cotyledon development, dehydrin mRNA and proteins accumulated in mid to late embryogenesis. Dehydrin proteins were some of the most actively synthesised at about the time of maximum fresh weight and represent about 2% of protein in mature cotyledons.

  12. 7 CFR 319.56-45 - Shelled garden peas from Kenya.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 5 2012-01-01 2012-01-01 false Shelled garden peas from Kenya. 319.56-45 Section 319... Shelled garden peas from Kenya. Garden peas (Pisum sativum) may be imported into the continental United... provisions of this subpart: (a) The peas must be shelled from the pod. (b) The peas must be washed in...

  13. 7 CFR 319.56-45 - Shelled garden peas from Kenya.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 5 2013-01-01 2013-01-01 false Shelled garden peas from Kenya. 319.56-45 Section 319... Shelled garden peas from Kenya. Garden peas (Pisum sativum) may be imported into the continental United... provisions of this subpart: (a) The peas must be shelled from the pod. (b) The peas must be washed in...

  14. 7 CFR 319.56-45 - Shelled garden peas from Kenya.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 5 2014-01-01 2014-01-01 false Shelled garden peas from Kenya. 319.56-45 Section 319... Shelled garden peas from Kenya. Garden peas (Pisum sativum) may be imported into the continental United... provisions of this subpart: (a) The peas must be shelled from the pod. (b) The peas must be washed in...

  15. 7 CFR 319.56-45 - Shelled garden peas from Kenya.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 5 2010-01-01 2010-01-01 false Shelled garden peas from Kenya. 319.56-45 Section 319... Shelled garden peas from Kenya. Garden peas (Pisum sativum) may be imported into the continental United... provisions of this subpart: (a) The peas must be shelled from the pod. (b) The peas must be washed in...

  16. 7 CFR 319.56-45 - Shelled garden peas from Kenya.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 5 2011-01-01 2011-01-01 false Shelled garden peas from Kenya. 319.56-45 Section 319... Shelled garden peas from Kenya. Garden peas (Pisum sativum) may be imported into the continental United... provisions of this subpart: (a) The peas must be shelled from the pod. (b) The peas must be washed...

  17. Stamina pistilloida, the Pea Ortholog of Fim and UFO, Is Required for Normal Development of Flowers, Inflorescences, and Leaves

    PubMed Central

    Taylor, Scott; Hofer, Julie; Murfet, Ian

    2001-01-01

    Isolation and characterization of two severe alleles at the Stamina pistilloida (Stp) locus reveals that Stp is involved in a wide range of developmental processes in the garden pea. The most severe allele, stp-4, results in flowers consisting almost entirely of sepals and carpels. Production of ectopic secondary flowers in stp-4 plants suggests that Stp is involved in specifying floral meristem identity in pea. The stp mutations also reduce the complexity of the compound pea leaf, and primary inflorescences often terminate prematurely in an aberrant sepaloid flower. In addition, stp mutants were shorter than their wild-type siblings due to a reduction in cell number in their internodes. Fewer cells were also found in the epidermis of the leaf rachis of stp mutants. Examination of the effects of stp-4 in double mutant combinations with af, tl, det, and veg2-2—mutations known to influence leaf, inflorescence, and flower development in pea—suggests that Stp function is independent of these genes. A synergistic interaction between weak mutant alleles at Stp and Uni indicated that these two genes act together, possibly to regulate primordial growth. Molecular analysis revealed that Stp is the pea homolog of the Antirrhinum gene Fimbriata (Fim) and of UNUSUAL FLORAL ORGANS (UFO) from Arabidopsis. Differences between Fim/UFO and Stp mutant phenotypes and expression patterns suggest that expansion of Stp activity into the leaf was an important step during evolution of the compound leaf in the garden pea. PMID:11158527

  18. Heat Stress Response in Pea Involves Interaction of Mitochondrial Nucleoside Diphosphate Kinase with a Novel 86-Kilodalton Protein1

    PubMed Central

    Escobar Galvis, Martha L.; Marttila, Salla; Håkansson, Gunilla; Forsberg, Jens; Knorpp, Carina

    2001-01-01

    In this work we have further characterized the first mitochondrial nucleoside diphosphate kinase (mtNDPK) isolated from plants. The mitochondrial isoform was found to be especially abundant in reproductive and young tissues. Expression of the pea (Pisum sativum L. cv Oregon sugarpod) mtNDPK was not affected by different stress conditions. However, the pea mtNDPK was found to interact with a novel 86-kD protein, which is de novo synthesized in pea leaves upon exposure to heat. Thus, we have evidence for the involvement of mtNDPK in mitochondrial heat response in pea in vivo. Studies on oligomerization revealed that mtNDPK was found in complexes of various sizes, corresponding to the sizes of e.g. hexamers, tetramers, and dimers, indicating flexibility in oligomerization. This flexibility, also found for other NDPK isoforms, has been correlated with the ability of this enzyme to interact with other proteins. We believe that the mtNDPK is involved in heat stress response in pea, possibly as a modulator of the 86-kD protein. PMID:11351071

  19. Dwarf mutations in grass pea (Lathyrus sativus L.): origin, morphology, inheritance and linkage studies.

    PubMed

    Talukdar, Dibyendu

    2009-08-01

    Induction of mutation has been used to create additional genetic variability in grass pea (Lathyrus sativus L.). During the ongoing investigations on different induced-morphological mutants, the author detected three types of dwarf mutants in grass pea. One mutant, designated as dwf1 type was earlier identified in colchicine-induced C2 generation of grass pea variety BioR-231 while the other two, designated as dwf2 and dwf3 were isolated in 250 Gy and 300 Gy gamma ray irradiated M2 progeny of variety 'BioR-231' and 'Hooghly Local', respectively. As compared to their parental varieties (controls), all the three mutants manifested stunted, erect and determinate stem, early maturity and tolerance to pod shattering habit. The mutants differed from each other, as well as with controls, in number of primary branches, nature of stipules and internodes, length of peduncle, leaflet and seed coat colour, seed yield and seed neurotoxin content. The three dwarf mutants were monogenically recessive and bred true in successive generations. F2 segregation pattern obtained from the crosses involving the three mutants indicated that dwarf mutation in grass pea was controlled by two independent non-allelic genes, assigned as df1 (for dwf1 type), df2 (for dwf2 type) and df3 (for dwf3 type), with the df1 locus being multiple allelic. Primary trisomic analyses revealed the presence of df1/df2 locus on the extra chromosome of trisomic type I, whereas df3 was located on the extra chromosome of type III. Linkage studies involving five other phenotypic markers suggested linked association of df1/df2 locus with lfc (leaflet colour) and wgn (winged internode) and df3 locus with cbl (seed coat colour). Both the loci; however, assorted independently with flower colour and stipule character. The dwarf types can be utilized as valuable tools for further cytogenetic research and breeding of grass pea.

  20. Antibody expressing pea seeds as fodder for prevention of gastrointestinal parasitic infections in chickens

    PubMed Central

    Zimmermann, Jana; Saalbach, Isolde; Jahn, Doreen; Giersberg, Martin; Haehnel, Sigrun; Wedel, Julia; Macek, Jeanette; Zoufal, Karen; Glünder, Gerhard; Falkenburg, Dieter; Kipriyanov, Sergey M

    2009-01-01

    Background Coccidiosis caused by protozoans of genus Eimeria is a chicken parasitic disease of great economical importance. Conventional disease control strategies depend on vaccination and prophylactic use of anticoccidial drugs. Alternative solution to prevent and treat coccidiosis could be provided by passive immunization using orally delivered neutralizing antibodies. We investigated the possibility to mitigate the parasitic infection by feeding poultry with antibody expressing transgenic crop seeds. Results Using the phage display antibody library, we generated a panel of anti-Eimeria scFv antibody fragments with high sporozoite-neutralizing activity. These antibodies were expressed either transiently in agrobacteria-infiltrated tobacco leaves or stably in seeds of transgenic pea plants. Comparison of the scFv antibodies purified either from tobacco leaves or from the pea seeds demonstrated no difference in their antigen-binding activity and molecular form compositions. Force-feeding experiments demonstrated that oral delivery of flour prepared from the transgenic pea seeds had higher parasite neutralizing activity in vivo than the purified antibody fragments isolated from tobacco. The pea seed content was found to protect antibodies against degradation by gastrointestinal proteases (>100-fold gain in stability). Ad libitum feeding of chickens demonstrated that the transgenic seeds were well consumed and not shunned. Furthermore, feeding poultry with shred prepared from the antibody expressing pea seeds led to significant mitigation of infection caused both by high and low challenge doses of Eimeria oocysts. Conclusion The results suggest that our strategy offers a general approach to control parasitic infections in production animals using cost-effective antibody expression in crop seeds affordable for the animal health market. PMID:19747368

  1. Plant waxy bloom on peas affects infection of pea aphids by Pandora neoaphidis.

    PubMed

    Duetting, Patrick S; Ding, Hongjian; Neufeld, Jeffrey; Eigenbrode, Sanford D

    2003-11-01

    This study examined the effects of the surface wax bloom of pea plants, Pisum sativum, on infection of pea aphids, Acyrthosiphon pisum, by the fungal pathogen Pandora neoaphidis. In prior field surveys, a higher proportion of P. neoaphidis-killed pea aphids (cadavers) had been observed on a pea line with reduced wax bloom, as compared with a sister line with normal surface wax bloom. Laboratory bioassays were conducted in order to examine the mechanisms. After plants of each line infested with aphids were exposed to similar densities of conidia, the rate of accumulation of cadavers on the reduced wax line was significantly greater than on the normal wax bloom line; at the end of the experiment (13d), the proportion of aphid cadavers on the reduced wax line was approximately four times that on the normal wax bloom line. When plants were exposed to conidia first and then infested with aphids, the rate of accumulation of cadavers was slightly but significantly greater on the reduced wax line, and infection at the end of the experiment (16d) did not differ between the lines. When aphids were exposed first and then released onto the plants, no differences in the proportion of aphid cadavers were observed between the pea lines. Greater infection of pea aphid on reduced wax peas appears to depend upon plants being exposed to inoculum while aphids are settled in typical feeding positions on the plant. Additional experiments demonstrated increased adhesion and germination by P. neoaphidis conidia to leaf surfaces of the reduced wax line as compared with normal wax line, and this could help explain the higher infection rate by P. neoaphidis on the reduced wax line. In bioassays using surface waxes extracted from the two lines, there was no effect of wax source on germination of P. neoaphidis conidia.

  2. Transcriptional profiling of the pea shoot apical meristem reveals processes underlying its function and maintenance

    PubMed Central

    Wong, Chui E; Bhalla, Prem L; Ottenhof, Harald; Singh, Mohan B

    2008-01-01

    Background Despite the importance of the shoot apical meristem (SAM) in plant development and organ formation, our understanding of the molecular mechanisms controlling its function is limited. Genomic tools have the potential to unravel the molecular mysteries of the SAM, and legume systems are increasingly being used in plant-development studies owing to their unique characteristics such as nitrogen fixation, secondary metabolism, and pod development. Garden pea (Pisum sativum) is a well-established classic model species for genetics studies that has been used since the Mendel era. In addition, the availability of a plethora of developmental mutants makes pea an ideal crop legume for genomics studies. This study aims to utilise genomics tools in isolating genes that play potential roles in the regulation of SAM activity. Results In order to identify genes that are differentially expressed in the SAM, we generated 2735 ESTs from three cDNA libraries derived from freshly micro-dissected SAMs from 10-day-old garden peas (Pisum sativum cv Torsdag). Custom-designed oligonucleotide arrays were used to compare the transcriptional profiles of pea SAMs and non-meristematic tissues. A total of 184 and 175 transcripts were significantly up- or down-regulated in the pea SAM, respectively. As expected, close to 61% of the transcripts down-regulated in the SAM were found in the public database, whereas sequences from the same source only comprised 12% of the genes that were expressed at higher levels in the SAM. This highlights the under-representation of transcripts from the meristematic tissues in the current public pea protein database, and demonstrates the utility of our SAM EST collection as an essential genetic resource for revealing further information on the regulation of this developmental process. In addition to unknowns, many of the up-regulated transcripts are known to encode products associated with cell division and proliferation, epigenetic regulation, auxin

  3. Aggressiveness Changes in Populations of Didymella pinodes over Winter and Spring Pea Cropping Seasons

    PubMed Central

    Laloi, G.; Montarry, J.; Guibert, M.; Andrivon, D.; Michot, D.

    2016-01-01

    ABSTRACT Ascochyta blight, caused by the necrotrophic ascomycete Didymella pinodes, is responsible for severe losses in winter and spring pea crops. Despite different climatic conditions, epidemics on winter and spring crops are due to a single population of D. pinodes, suggesting gene flow either between the two crops or from reservoir sources during the cropping season. This should lead to similar pathogenicity characteristics in isolates sampled from the two crops. However, these hypotheses have never been formally tested. We therefore sampled a total of 520 D. pinodes strains throughout a growing season from winter and spring pea plots (WP and SP, respectively) and from winter and spring trap plants (TWP and TSP). Amplified fragment length polymorphism (AFLP) markers revealed high genetic diversity within subpopulations, whereas pathogenicity tests showed that mean aggressiveness increases over the course of an epidemic. These results support the idea that alloinoculum contributes to the carryover of epidemics between winter and spring crops and that the most aggressive isolates are selected as an epidemic progresses. IMPORTANCE Ascochyta blight, caused by Didymella pinodes, is responsible for severe losses in pea crops. While previous studies have shown that ascochyta blight epidemics on winter and spring crops are due to a single population of D. pinodes, suggesting that isolates from the two crops present similar pathogenicity characteristics, that hypothesis have never been tested. Genetic analysis of subpopulations sampled throughout a growing season from winter and spring pea plots revealed high genetic diversity within subpopulations, whereas pathogenicity tests showed that mean aggressiveness increases over the course of an epidemic. PMID:27208102

  4. Response of Pea Varieties to Damage Degree of Pea Weevil, Bruchus pisorum L.

    PubMed Central

    Nikolova, Ivelina Mitkova

    2016-01-01

    A study was conducted to determine the response of five pea varieties (Pisum sativum L.) to damage degree of Bruchus pisorum: Glyans, Modus, Kamerton, and Svit (Ukrainian cultivars) and Pleven 4 (Bulgarian cultivar). The seeds were classified into three types: healthy seeds (type 1), damaged seeds with parasitoid emergence hole (type 2), and damaged seeds with bruchid emergence hole (type 3) and they were sown. It was found that the weight of 1000 seeds did not affect the field germination of the pea varieties. Healthy and damaged seeds with parasitoid emergence holes (first and second seed types) provide a very good opportunity for growth and development while plants from damaged seeds with bruchid emergence holes had poor germination and vigor and low productivity. These seeds cannot provide the creation of well-garnished seeding and stable crop yields. Among tested varieties, the Ukrainian variety Glyans had considerably higher seed weight, field germination, and index germination and weak egg-laying activity of B. pisorum compared to others. Use of spring pea cultivars that are weakly preferred by the pea weevil in breeding programs would reduce losses due to pea weevil and provide an environmentally safer option to its control. PMID:27042379

  5. Active galactic nuclei

    PubMed Central

    Fabian, Andrew C.

    1999-01-01

    Active galactic nuclei are the most powerful, long-lived objects in the Universe. Recent data confirm the theoretical idea that the power source is accretion into a massive black hole. The common occurrence of obscuration and outflows probably means that the contribution of active galactic nuclei to the power density of the Universe has been generally underestimated. PMID:10220363

  6. Pea, Pisum sativum, and Its Anticancer Activity

    PubMed Central

    Rungruangmaitree, Runchana; Jiraungkoorskul, Wannee

    2017-01-01

    Pisum sativum (Family: Fabaceae), as known as green pea or garden pea, has long been important in diet due to its content of fiber, protein, starch, trace elements, and many phytochemical substances. It has been shown to possess antibacterial, antidiabetic, antifungal, anti-inflammatory, antihypercholesterolemia, and antioxidant activities and also shown anticancer property. Its nonnutritive biologically active components include alkaloids, flavonoids, glycosides, isoflavones, phenols, phytosterols, phytic acid, protease inhibitors, saponins, and tannins. This plant is rich in apigenin, hydroxybenzoic, hydroxycinnamic, luteolin, and quercetin, all of which have been reported to contribute to its remedial properties including anticarcinogenesis property. Based on established literature on the anticancer property of P. sativum and possible mode of action, this review article has focused to demonstrate that P. sativum could be further explored for the development of anticancer treatment. PMID:28503053

  7. Pea, Pisum sativum, and Its Anticancer Activity.

    PubMed

    Rungruangmaitree, Runchana; Jiraungkoorskul, Wannee

    2017-01-01

    Pisum sativum (Family: Fabaceae), as known as green pea or garden pea, has long been important in diet due to its content of fiber, protein, starch, trace elements, and many phytochemical substances. It has been shown to possess antibacterial, antidiabetic, antifungal, anti-inflammatory, antihypercholesterolemia, and antioxidant activities and also shown anticancer property. Its nonnutritive biologically active components include alkaloids, flavonoids, glycosides, isoflavones, phenols, phytosterols, phytic acid, protease inhibitors, saponins, and tannins. This plant is rich in apigenin, hydroxybenzoic, hydroxycinnamic, luteolin, and quercetin, all of which have been reported to contribute to its remedial properties including anticarcinogenesis property. Based on established literature on the anticancer property of P. sativum and possible mode of action, this review article has focused to demonstrate that P. sativum could be further explored for the development of anticancer treatment.

  8. Reactive oxygen species in programmed death of pea guard cells.

    PubMed

    Samuilov, V D; Kiselevsky, D B; Shestak, A A; Nesov, A V; Vasil'ev, L A

    2008-10-01

    Hydrogen peroxide potentiates CN(-)-induced apoptosis of guard cells recorded as destruction of cell nuclei in the epidermis from pea leaves. A still stronger effect was exerted by the addition of H2O2 and NADH, which are the substrates of the plant cell wall peroxidase producing O2*- coupled to the oxidation of NADH. The CN(-)-or (CN(-) + H2O2)-induced destruction of guard cell nuclei was completely removed by nitroblue tetrazolium (NBT) oxidizing O2*- and preventing there-by the subsequent generation of H2O2. The reduced NBT was deposited in the cells as formazan crystals. Cyanide-induced apoptosis was diminished by mannitol and ethanol, which are OH* traps. The dyes Rose Bengal (RB) and tetramethylrhodamine ethyl ester (TMRE) photosensitizing singlet oxygen production suppressed the CN(-)-induced destruction of the cell nuclei in the light. This suppression was removed by exogenous NADH, which reacts with 1O2 yielding O2*-. Incubation of leaf slices with RB in the light lowered the photosynthetic O2 evolution rate and induced the permeability of guard cells for propidium iodide, which cannot pass across intact membranes. Inhibition of photosynthetic O2 evolution by 3-(3',4'-dichlorophenyl)-1,1-dimethylurea or bromoxynil prevented CN(-)-induced apoptosis of guard cells in the light but not in the dark. RB in combination with exogenous NADH caused H2O2 production that was sensitive to NBT and estimated from dichlorofluorescein (DCF) fluorescence. Data on NBT reduction and DCF and TMRE fluorescence obtained using a confocal microscope and data on the NADH-dependent H2O2 production are indicative of generation of reactive oxygen species in the chloroplasts, mitochondria, and nuclear region of guard cells as well as with participation of apoplastic peroxidase. Cyanide inhibited generation of reactive oxygen species in mitochondria and induced their generation in chloroplasts. The results show that H2O2, OH*, and O2*- resources utilized for H2O2 production are

  9. Genomic Tools in Pea Breeding Programs: Status and Perspectives

    PubMed Central

    Tayeh, Nadim; Aubert, Grégoire; Pilet-Nayel, Marie-Laure; Lejeune-Hénaut, Isabelle; Warkentin, Thomas D.; Burstin, Judith

    2015-01-01

    Pea (Pisum sativum L.) is an annual cool-season legume and one of the oldest domesticated crops. Dry pea seeds contain 22–25% protein, complex starch and fiber constituents, and a rich array of vitamins, minerals, and phytochemicals which make them a valuable source for human consumption and livestock feed. Dry pea ranks third to common bean and chickpea as the most widely grown pulse in the world with more than 11 million tons produced in 2013. Pea breeding has achieved great success since the time of Mendel's experiments in the mid-1800s. However, several traits still require significant improvement for better yield stability in a larger growing area. Key breeding objectives in pea include improving biotic and abiotic stress resistance and enhancing yield components and seed quality. Taking advantage of the diversity present in the pea genepool, many mapping populations have been constructed in the last decades and efforts have been deployed to identify loci involved in the control of target traits and further introgress them into elite breeding materials. Pea now benefits from next-generation sequencing and high-throughput genotyping technologies that are paving the way for genome-wide association studies and genomic selection approaches. This review covers the significant development and deployment of genomic tools for pea breeding in recent years. Future prospects are discussed especially in light of current progress toward deciphering the pea genome. PMID:26640470

  10. Adipocyte nuclei captured from VAT and SAT.

    PubMed

    Ambati, Suresh; Yu, Ping; McKinney, Elizabeth C; Kandasamy, Muthugapatti K; Hartzell, Diane; Baile, Clifton A; Meagher, Richard B

    2016-01-01

    Obesity-related comorbidities are thought to result from the reprogramming of the epigenome in numerous tissues and cell types, and in particular, mature adipocytes within visceral and subcutaneous adipose tissue, VAT and SAT. The cell-type specific chromatin remodeling of mature adipocytes within VAT and SAT is poorly understood, in part, because of the difficulties of isolating and manipulating large fragile mature adipocyte cells from adipose tissues. We constructed MA-INTACT (Mature Adipocyte-Isolation of Nuclei TAgged in specific Cell Types) mice using the adiponectin (ADIPOQ) promoter (ADNp) to tag the surface of mature adipocyte nuclei with a reporter protein. The SUN1mRFP1Flag reporter is comprised of a fragment of the nuclear transmembrane protein SUN1, the fluorescent protein mRFP1, and three copies of the Flag epitope tag. Mature adipocyte nuclei were rapidly and efficiently immuno-captured from VAT and SAT (MVA and MSA nuclei, respectively), of MA-INTACT mice. MVA and MSA nuclei contained 1,000 to 10,000-fold higher levels of adipocyte-specific transcripts, ADIPOQ, PPARg2, EDNRB, and LEP, relative to uncaptured nuclei, while the latter expressed higher levels of leukocyte and endothelial cell markers IKZF1, RETN, SERPINF1, SERPINE1, ILF3, and TNFA. MVA and MSA nuclei differentially expressed several factors linked to adipogenesis or obesity-related health risks including CEBPA, KLF2, RETN, SERPINE1, and TNFA. The various nuclear populations dramatically differentially expressed transcripts encoding chromatin remodeler proteins regulating DNA cytosine methylation and hydroxymethylation (TETs, DNMTs, TDG, GADD45s) and nucleosomal histone modification (ARID1A, KAT2B, KDM4A, PRMT1, PRMT5, PAXIP1). Remarkably, MSA and MVA nuclei expressed 200 to 1000-fold higher levels of thermogenic marker transcripts PRDM16 and UCP1. The MA-INTACT mouse enables a simple way to perform cell-type specific analysis of highly purified mature adipocyte nuclei from VAT and SAT

  11. Molecular cloning of isoflavone reductase from pea (Pisum sativum L.): evidence for a 3R-isoflavanone intermediate in (+)-pisatin biosynthesis.

    PubMed

    Paiva, N L; Sun, Y; Dixon, R A; VanEtten, H D; Hrazdina, G

    1994-08-01

    Isoflavone reductase (IFR) reduces achiral isoflavones to chiral isoflavanones during the biosynthesis of chiral pterocarpan phytoalexins. A cDNA clone for IFR from pea (Pisum sativum) was isolated using the polymerase chain reaction and expressed in Escherichia coli. Analysis of circular dichroism (CD) spectra of the reduction product sophorol obtained using the recombinant enzyme indicated that the isoflavanone possessed the 3R stereochemistry, in contrast to previous reports indicating a 3S-isoflavanone as the product of the pea IFR. Analysis of CD spectra of sophorol produced using enzyme extracts of CuCl2-treated pea seedlings confirmed the 3R stereochemistry. Thus, the stereochemistry of the isoflavanone intermediate in (+)-pisatin biosynthesis in pea is the same as that in (-)-medicarpin biosynthesis in alfalfa, although the final pterocarpans have the opposite stereochemistry. At the amino acid level the pea IFR cDNA was 91.8 and 85.2% identical to the IFRs from alfalfa and chickpea, respectively. IFR appears to be encoded by a single gene in pea. Its transcripts are highly induced in CuCl2-treated seedlings, consistent with the appearance of IFR enzyme activity and pisatin accumulation.

  12. [THE EFFECT OF ACID RAIN ON ULTRASTRUCTURE AND FUNCTIONAL PARAMETERS OF PHOTOSYNTHETIC APPARATUS OF PEA LEAVES].

    PubMed

    Polishchuk, A V; Vodka, M V; Belyavskaya, N A; Khomochkin, A P; Zolotareva, E K

    2016-01-01

    The effects of simulated acid rain (SAR) on the ultrastructure and functional parameters of the photosynthetic apparatus were studied using 14-day-old pea leaves as test system. Pea plants were sprayed with an aqueous solution containing NaNO₃(0.2 mM) and Na₂SO₄(0.2 mM) (pH 5.6, a control variant), or with the same solution, which was acidified to pH 2.5 (acid variant). Functional characteristics were determined by chlorophyll fluorescence analysis. Acid rain application caused reduction in the efficiency of the photosynthetic electron transport by 25%, which was accompanied by an increase by 85% in the quantum yield of thermal dissipation of excess light quanta. Ultrastructural changes in chloroplast were registered by transmission electron microscopy (TEM) after two days of the SAR-treatment of pea leaves. In this case, the changes in the structure of grana, heterogeneity of thylakoids packaging in granum, namely, the increase of intra-thylakoid gaps and thickness of granal thylakoids compared to the control were found. The migration of protein complexes in thylakoid membranes of chloroplasts isolated from leaves treated with SAR was suppressed. It was shown also that carbonic anhydrase activity was inhibited in chloroplast preparations isolated from SAR-treated pea leaves. We proposed a hypothesis on the possible inactivation of thylakoid carbonic anhydrase under SAR and its involvement in the inhibition of photochemical activity of chloroplasts. The data obtained allows to suggest that acid rains negatively affect the photosynthetic apparatus disrupting the membrane system of chloroplast.

  13. Efficient intergeneric fusion of pea (Pisum sativum L.) and grass pea (Lathyrus sativus L.) protoplasts.

    PubMed

    Durieu, P; Ochatt, S J

    2000-07-01

    Large numbers of viable protoplasts of pea (Pisum sativum) and grass pea (Lathyrus sativus) were efficiently and reproducibly obtained and, for the first time, fused. Different procedures for fusion were compared, based either on electrofusion (750, 1000, 1250 or 1500 V cm(-1)), or on the use of macro or micromethods with a polyethylene glycol (PEG 6000 or PEG 1540), or a glycine/high pH solution. Over 10% of viable heterokaryons were obtained, with PEG as the most efficient and reproducible agent for protoplast fusion (>20% of viable heterokaryons). Both the division of heterokaryons and the formation of small calluses were observed.

  14. Gibberellin (GA3) enhances cell wall invertase activity and mRNA levels in elongating dwarf pea (Pisum sativum) shoots

    NASA Technical Reports Server (NTRS)

    Wu, L. L.; Mitchell, J. P.; Cohn, N. S.; Kaufman, P. B.

    1993-01-01

    The invertase (EC 3.2.1.26) purified from cell walls of dwarf pea stems to homogeneity has a molecular mass of 64 kilodaltons (kD). Poly(A)+RNA was isolated from shoots of dwarf pea plants, and a cDNA library was constructed using lambda gt11 as an expression vector. The expression cDNA library was screened with polyclonal antibodies against pea cell wall invertase. One invertase cDNA clone was characterized as a full-length cDNA with 1,863 base pairs. Compared with other known invertases, one homologous region in the amino acid sequence was found. The conserved motif, Asn-Asp-Pro-Asn-Gly, is located near the N-terminal end of invertase. Northern blot analysis showed that the amount of invertase mRNA (1.86 kb) was rapidly induced to a maximal level 4 h after GA3 treatment, then gradually decreased to the control level. The mRNA level at 4 h in GA3-treated peas was fivefold higher than that of the control group. The maximal increase in activity of pea cell wall invertase elicited by GA3 occcured at 8 h after GA3 treatment. This invertase isoform was shown immunocytochemically to be localized in the cell walls, where a 10-fold higher accumulation occurred in GA3-treated tissue compared with control tissue. This study indicates that the expression of the pea shoot cell-wall invertase gene could be regulated by GA3 at transcriptional and/or translational levels.

  15. Gibberellin (GA3) enhances cell wall invertase activity and mRNA levels in elongating dwarf pea (Pisum sativum) shoots

    NASA Technical Reports Server (NTRS)

    Wu, L. L.; Mitchell, J. P.; Cohn, N. S.; Kaufman, P. B.

    1993-01-01

    The invertase (EC 3.2.1.26) purified from cell walls of dwarf pea stems to homogeneity has a molecular mass of 64 kilodaltons (kD). Poly(A)+RNA was isolated from shoots of dwarf pea plants, and a cDNA library was constructed using lambda gt11 as an expression vector. The expression cDNA library was screened with polyclonal antibodies against pea cell wall invertase. One invertase cDNA clone was characterized as a full-length cDNA with 1,863 base pairs. Compared with other known invertases, one homologous region in the amino acid sequence was found. The conserved motif, Asn-Asp-Pro-Asn-Gly, is located near the N-terminal end of invertase. Northern blot analysis showed that the amount of invertase mRNA (1.86 kb) was rapidly induced to a maximal level 4 h after GA3 treatment, then gradually decreased to the control level. The mRNA level at 4 h in GA3-treated peas was fivefold higher than that of the control group. The maximal increase in activity of pea cell wall invertase elicited by GA3 occcured at 8 h after GA3 treatment. This invertase isoform was shown immunocytochemically to be localized in the cell walls, where a 10-fold higher accumulation occurred in GA3-treated tissue compared with control tissue. This study indicates that the expression of the pea shoot cell-wall invertase gene could be regulated by GA3 at transcriptional and/or translational levels.

  16. Exotic Light Nuclei

    ERIC Educational Resources Information Center

    Cerny, Joseph; Poskanzer, Arthur M.

    1978-01-01

    Among the light elements, nuclei with unequal numbers of protons and neutrons are highly unstable. Some survive just long enough to be detected and exhibit unusual regimes of radioactive decay. ( Autor/MA)

  17. Exotic Light Nuclei

    ERIC Educational Resources Information Center

    Cerny, Joseph; Poskanzer, Arthur M.

    1978-01-01

    Among the light elements, nuclei with unequal numbers of protons and neutrons are highly unstable. Some survive just long enough to be detected and exhibit unusual regimes of radioactive decay. ( Autor/MA)

  18. Observations of cometary nuclei

    NASA Astrophysics Data System (ADS)

    A'Hearn, M. F.

    Attempts to observe cometary nuclei and to determine fundamental physical parameters relevant to the relationship between comets and asteroids are reviewed. It has been found that cometary nuclei, at least of periodic comets, are bigger and blacker than generally thought as recently as five years ago. Geometric albedos may be typically three percent and typical radii are probably of order 5 km. Nuclei of periodic comets are probably highly prolate unless they are both oblate and rotating about one of the major axes. P/Halley images provide convincing evidence of the existence of mantles discussed in many models. Numerous pieces of evidence suggest a connection between cometary nuclei and A-A asteroids of types D and C.

  19. Chapter 8. Genetic Adjustment to Changing Climates: Pea

    USDA-ARS?s Scientific Manuscript database

    Peas are adapted to cool, semi-arid to sub-humid growing conditions and although they are widely grown throughout the world, the best performance is realized in the cool, relatively dry areas of the mid-latitudes. The literature to date on the potential effects of climate change specifically on pea ...

  20. Pea (Pisum sativum L.) in the genomics era

    USDA-ARS?s Scientific Manuscript database

    Pea (Pisum sativum L.) was the original model organism for Mendel´s discovery of the laws of inheritance, making it the foundation of modern plant genetics. However, subsequent progress in pea genomics has lagged behind many other plant species, largely as a consequence of its low multiplication rat...

  1. Appearance of Three Chloroplast Isoenzymes in Dark-grown Pea Plants and Pea Seeds 12

    PubMed Central

    Park, Kyung Eun Yoon; Anderson, Louise E.

    1973-01-01

    Activity peaks characteristic of the chloroplastic Calvin cycle enzymes triose-phosphate isomerase, ribose 5-phosphate isomerase, and fructose 1,6-diphosphate aldolase are found in isoelectric focusing patterns of dark-grown pea (Pisum sativum) seedlings and seeds. Apparently, in this higher plant these three chloroplastic isoenzymes can be formed in the absence of light and of chloroplast formation. PMID:16658311

  2. Fibril formation from pea protein and subsequent gel formation.

    PubMed

    Munialo, Claire Darizu; Martin, Anneke H; van der Linden, Erik; de Jongh, Harmen H J

    2014-03-19

    The objective of this study was to characterize fibrillar aggregates made using pea proteins, to assemble formed fibrils into protein-based gels, and to study the rheological behavior of these gels. Micrometer-long fibrillar aggregates were observed after pea protein solutions had been heated for 20 h at pH 2.0. Following heating of pea proteins, it was observed that all of the proteins were hydrolyzed into peptides and that 50% of these peptides were assembled into fibrils. Changes on a structural level in pea proteins were studied using circular dichroism, transmission electron microscopy, and particle size analysis. During the fibril assembly process, an increase in aggregate size was observed, which coincided with an increase in thioflavin T binding, indicating the presence of β-sheet aggregates. Fibrils made using pea proteins were more branched and curly. Gel formation of preformed fibrils was induced by slow acidification from pH 7.0 to a final pH of around pH 5.0. The ability of pea protein-based fibrillar gels to fracture during an amplitude sweep was comparable to those of soy protein and whey protein-based fibrillar gels, although gels prepared from fibrils made using pea protein and soy protein were weaker than those of whey protein. The findings show that fibrils can be prepared from pea protein, which can be incorporated into protein-based fibrillar gels.

  3. Plant characteristics and growth parameters of vegetable pigeon pea cultivars

    USDA-ARS?s Scientific Manuscript database

    Pigeon pea is an important crop in dry land and semi-arid regions and is a supplementary source of dietary protein for the resource-constrained farmers. The aim of this research was to evaluate growth parameters of twelve vegetable pigeon pea genotypes at two locations in Eastern Kenya. The number o...

  4. Development of genetic resources for association mapping in pea

    USDA-ARS?s Scientific Manuscript database

    A previous study of the allelic diversity of the USDA pea core collection resequenced ten loci of 32 diverse pea germplasm accessions from which SNP assays were designed for Agpl1, AspS2, Cwi1, DRR206c, PPlike, PsAS2, SOD9, Thaumatin1, PsLD, and Viola2 (Aubert et al. 2006; Gilpin et al 1997; Hecht ...

  5. Additional pea EST-SSR markers for comparative mapping in pea (Pisum sativum L.)

    USDA-ARS?s Scientific Manuscript database

    The availability of a large set of molecular markers is an excellent resource for both applied and basic research for any organism. The current consensus molecular map of pea is 1,430 cM and contains 239 simple sequence repeat markers. The goal was to identify and test unique gene-based simple sequ...

  6. The Effect of Orobanche crenata Infection Severity in Faba Bean, Field Pea, and Grass Pea Productivity

    PubMed Central

    Fernández-Aparicio, Mónica; Flores, Fernando; Rubiales, Diego

    2016-01-01

    Broomrape weeds (Orobanche and Phelipanche spp.) are root holoparasites that feed off a wide range of important crops. Among them, Orobanche crenata attacks legumes complicating their inclusion in cropping systems along the Mediterranean area and West Asia. The detrimental effect of broomrape parasitism in crop yield can reach up to 100% depending on infection severity and the broomrape-crop association. This work provides field data of the consequences of O. crenata infection severity in three legume crops, i.e., faba bean, field pea, and grass pea. Regression functions modeled productivity losses and revealed trends in dry matter allocation in relation to infection severity. The host species differentially limits parasitic sink strength indicating different levels of broomrape tolerance at equivalent infection severities. Reductions in host aboveground biomass were observed starting at low infection severity and half maximal inhibitory performance was predicted as 4.5, 8.2, and 1.5 parasites per faba bean, field pea, and grass pea plant, respectively. Reductions in host biomass occurred in both vegetative and reproductive organs, the latter resulting more affected. The increase of resources allocated within the parasite was concomitant to reduction of host seed yield indicating that parasite growth and host reproduction compete directly for resources within a host plant. However, the parasitic sink activity does not fully explain the total host biomass reduction because combined biomass of host–parasite complex was lower than the biomass of uninfected plants. In grass pea, the seed yield was negligible at severities higher than four parasites per plant. In contrast, faba bean and field pea sustained low but significant seed production at the highest infection severity. Data on seed yield and seed number indicated that the sensitivity of field pea to O. crenata limited the production of grain yield by reducing seed number but maintaining seed size. In contrast

  7. Purification and Characterization of a Lectin from Green Split Peas (Pisum sativum).

    PubMed

    Ng, Tzi Bun; Chan, Yau Sang; Ng, Charlene Cheuk Wing; Wong, Jack Ho

    2015-11-01

    Lectins have captured the attention of a large number of researchers on account of their various exploitable activities, including antitumor, immunomodulatory, antifungal, as well as HIV reverse transcriptase inhibitory activities. A mannose/glucose-specific lectin was isolated from green split peas (a variety of Pisum sativum) and characterized. The purification step involved anion-exchange chromatography on a DEAE-cellulose column, cation-exchange chromatography on an SP-Sepharose column, and gel filtration by fast protein liquid chromatography (FPLC) on Superdex 200. The purified lectin had a native molecular mass of around 50 kDa as determined by size exclusion chromatography. It appeared as a heterotetramer, composed of two distinct polypeptide bands with a molecular mass of 6 and 19 kDa, respectively, in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The N-terminal sequence of green split pea lectin shows some degree of homology compared to lectins from other legume species. Its hemagglutinating activity was inhibited by glucose, mannose, and sucrose, and attenuated at pH values higher than 12 or lower than 3. Hemagglutinating activity was preserved at temperatures lower than 80 °C. The lectin did not show antifungal activity toward fungi including Fusarium oxysporum, Botrytis cinerea, and Mycosphaerella arachidicola. Green split pea lectin showed a mitogenic effect toward murine splenocytes and could inhibit the activity of HIV-1 reverse transcriptase.

  8. Cloning and functional characterization of a homoglutathione synthetase from pea nodules.

    PubMed

    Iturbe-Ormaetxe, Iñaki; Heras, Begoña; Matamoros, Manuel A; Ramos, Javier; Moran, Jose F; Becana, Manuel

    2002-05-01

    The thiol tripeptide glutathione (GSH; gammaGlu-Cys-Gly) is very abundant in legume nodules where it performs multiple functions that are critical for optimal nitrogen fixation. Some legume nodules contain another tripeptide, homoglutathione (hGSH; gammaGlu-Cys-betaAla), in addition to or instead of GSH. We have isolated from a pea (Pisum sativum L.) nodule library a cDNA, GSHS2, that is expressed in nodules but not in leaves. This cDNA was overexpressed in insect cells and its protein product was identified as a highly active and specific hGSH synthetase. The enzyme, the first of this type to be completely purified, is predicted to be a homodimeric cytosolic protein. It shows a specific activity of 3400 nmol hGSH min-1 mg-1 protein with a standard substrate concentration (5 mM beta-alanine) and Km values of 1.9 mM for beta-alanine and 104 mM for glycine. The specificity constant (Vmax/Km) shows that the pure enzyme is 57.3-fold more specific for beta-alanine than for glycine. Southern blot analysis revealed that the gene is present as a single copy in the pea genome and that there are homologous genes in other legumes. We conclude that the synthesis of hGSH in pea nodules is catalysed by a specific hGSH synthetase and not by a GSH synthetase with broad substrate specificity.

  9. The LKA gene is a BRASSINOSTEROID INSENSITIVE 1 homolog of pea.

    PubMed

    Nomura, Takahito; Bishop, Gerard J; Kaneta, Tsuyoshi; Reid, James B; Chory, Joanne; Yokota, Takao

    2003-11-01

    Brassinosteroids (BRs) are growth-promoting plant steroid hormones, and in garden pea (Pisum sativum L.), the lka mutant is defective in BR perception. Here, we show that LKA encodes P. sativum BRI1 (PsBRI1), a homolog of BRI1, which is the Arabidopsis leucine-rich repeat receptor-like kinase/BR receptor. PsBRI1 was isolated by screening a pea cDNA library using Arabidopsis BRI1 cDNA as the probe. PsBRI1 is predicted to encode a 1188-amino-acid protein that has 78% similarity with Arabidopsis BRI1. Sequence analysis of PsBRI1 in the lka mutant led to the identification of a missense mutation that converts the highly conserved aspartic acid residue to asparagine, which is located in the leucine-rich repeat, just before the island domain that may bind BR or a BR-protein complex. The mutation identified in PsBRI1 co-segregated with the semi-erectoide lka phenotype. Transcript analysis of LKA/PsBRI1 indicates that it is ubiquitously expressed in pea and that the expression was downregulated by exogenous BR. The lka mutant was then utilized in further studies to analyze the independent actions of BR and gibberellin (GA) through the characterization of BR response on GA mutants and GA response on BR mutants.

  10. Studies on the Control of Ascochyta Blight in Field Peas (Pisum sativum L.) Caused by Ascochyta pinodes in Zhejiang Province, China

    PubMed Central

    Liu, Na; Xu, Shengchun; Yao, Xiefeng; Zhang, Guwen; Mao, Weihua; Hu, Qizan; Feng, Zhijuan; Gong, Yaming

    2016-01-01

    Ascochyta blight, an infection caused by a complex of Ascochyta pinodes, Ascochyta pinodella, Ascochyta pisi, and/or Phoma koolunga, is a destructive disease in many field peas (Pisum sativum L.)-growing regions, and it causes significant losses in grain yield. To understand the composition of fungi associated with this disease in Zhejiang Province, China, a total of 65 single-pycnidiospore fungal isolates were obtained from diseased pea samples collected from 5 locations in this region. These isolates were identified as Ascochyta pinodes by molecular techniques and their morphological and physiological characteristics. The mycelia of ZJ-1 could penetrate pea leaves across the stomas, and formed specific penetration structures and directly pierced leaves. The resistance level of 23 available pea cultivars was tested against their representative isolate A. pinodes ZJ-1 using the excised leaf-assay technique. The ZJ-1 mycelia could penetrate the leaves of all tested cultivars, and they developed typical symptoms, which suggested that all tested cultivars were susceptible to the fungus. Chemical fungicides and biological control agents were screened for management of this disease, and their efficacies were further determined. Most of the tested fungicides (11 out of 14) showed high activity toward ZJ-1 with EC50 < 5 μg/mL. Moreover, fungicides, including tebuconazole, boscalid, iprodione, carbendazim, and fludioxonil, displayed more than 80% disease control efficacy under the recorded conditions. Three biocontrol strains of Bacillus sp. and one of Pantoea agglomerans were isolated from pea-related niches and significantly reduced the severity of disease under greenhouse and field conditions. To our knowledge, this is the first study on ascochyta blight in field peas, and results presented here will be useful for controlling the disease in this area. PMID:27148177

  11. Studies on the Control of Ascochyta Blight in Field Peas (Pisum sativum L.) Caused by Ascochyta pinodes in Zhejiang Province, China.

    PubMed

    Liu, Na; Xu, Shengchun; Yao, Xiefeng; Zhang, Guwen; Mao, Weihua; Hu, Qizan; Feng, Zhijuan; Gong, Yaming

    2016-01-01

    Ascochyta blight, an infection caused by a complex of Ascochyta pinodes, Ascochyta pinodella, Ascochyta pisi, and/or Phoma koolunga, is a destructive disease in many field peas (Pisum sativum L.)-growing regions, and it causes significant losses in grain yield. To understand the composition of fungi associated with this disease in Zhejiang Province, China, a total of 65 single-pycnidiospore fungal isolates were obtained from diseased pea samples collected from 5 locations in this region. These isolates were identified as Ascochyta pinodes by molecular techniques and their morphological and physiological characteristics. The mycelia of ZJ-1 could penetrate pea leaves across the stomas, and formed specific penetration structures and directly pierced leaves. The resistance level of 23 available pea cultivars was tested against their representative isolate A. pinodes ZJ-1 using the excised leaf-assay technique. The ZJ-1 mycelia could penetrate the leaves of all tested cultivars, and they developed typical symptoms, which suggested that all tested cultivars were susceptible to the fungus. Chemical fungicides and biological control agents were screened for management of this disease, and their efficacies were further determined. Most of the tested fungicides (11 out of 14) showed high activity toward ZJ-1 with EC50 < 5 μg/mL. Moreover, fungicides, including tebuconazole, boscalid, iprodione, carbendazim, and fludioxonil, displayed more than 80% disease control efficacy under the recorded conditions. Three biocontrol strains of Bacillus sp. and one of Pantoea agglomerans were isolated from pea-related niches and significantly reduced the severity of disease under greenhouse and field conditions. To our knowledge, this is the first study on ascochyta blight in field peas, and results presented here will be useful for controlling the disease in this area.

  12. An image processing pipeline to detect and segment nuclei in muscle fiber microscopic images.

    PubMed

    Guo, Yanen; Xu, Xiaoyin; Wang, Yuanyuan; Wang, Yaming; Xia, Shunren; Yang, Zhong

    2014-08-01

    Muscle fiber images play an important role in the medical diagnosis and treatment of many muscular diseases. The number of nuclei in skeletal muscle fiber images is a key bio-marker of the diagnosis of muscular dystrophy. In nuclei segmentation one primary challenge is to correctly separate the clustered nuclei. In this article, we developed an image processing pipeline to automatically detect, segment, and analyze nuclei in microscopic image of muscle fibers. The pipeline consists of image pre-processing, identification of isolated nuclei, identification and segmentation of clustered nuclei, and quantitative analysis. Nuclei are initially extracted from background by using local Otsu's threshold. Based on analysis of morphological features of the isolated nuclei, including their areas, compactness, and major axis lengths, a Bayesian network is trained and applied to identify isolated nuclei from clustered nuclei and artifacts in all the images. Then a two-step refined watershed algorithm is applied to segment clustered nuclei. After segmentation, the nuclei can be quantified for statistical analysis. Comparing the segmented results with those of manual analysis and an existing technique, we find that our proposed image processing pipeline achieves good performance with high accuracy and precision. The presented image processing pipeline can therefore help biologists increase their throughput and objectivity in analyzing large numbers of nuclei in muscle fiber images.

  13. Ethylene effects in pea stem tissue

    SciTech Connect

    Steen, D.A.; Chadwick, A.V.

    1981-01-01

    The marked effects of ethylene on pea stem growth have been investigated. Low temperatures and colchicine, both known microtubule depolymerization agents, reverse the effects of ethylene in straight growth tests. Low temperature (6 C) also profoundly reduces the effects of gas in terms of swelling, hook curvature, and horizontal mutation. Deuterium oxide, an agent capable of rigidifying microtubular structure, mimics the effects of ethylene. Electron microscopy shows that microtubule orientation is strikingly altered by ethylene. These findings indicate that some of the ethylene responses may be due to a stabilizing effect on microtubules in plant cells.

  14. Protein gels and emulsions from mixtures of Cape hake and pea proteins.

    PubMed

    Tomé, Ana Sofia; Pires, Carla; Batista, Irineu; Sousa, Isabel; Raymundo, Anabela

    2015-01-01

    Portioning of frozen fish generates by-products such as fish 'sawdust' and cut-offs which can be further processed into protein concentrates and isolates. The objective of the present work was to produce gels and emulsions using recovered Cape hake protein powder (HPP). In previous works, the structures of the gels produced by HPP were found to be strong, with a high rubbery character. In this work, the addition of commercial pea proteins (PPC) to HPP gels and emulsions was studied. Physical properties of gels and emulsions prepared with different proportions of mixtures of PPC and HPP were evaluated. In general, gels and emulsions showed high values for whiteness and, as expected, the higher content of HPP in the protein mixtures led to higher firmness values of the gels. The gel network was rapidly formed upon heating due to the fish protein macromolecules and further reinforced by the pea protein macromolecules when cooled to 5 °C. Both visco-elastic parameters, storage and loss moduli, of the produced gels increased with the HPP proportion in the protein mixtures, corresponding to more structured systems. For the emulsions, two different pH environments were studied: 3.8 and 7.0. At neutral pH a synergy was found between the vegetable and fish protein, which is not so strong when pH is lowered to 3.8, near the isoelectric point of pea proteins (pI = 4.5). This evidence was supported by the results from the texture measurements, viscosity and visco-elastic parameters. Gels made from Cape hake proteins showed a softer texture and were less rubbery with the addition of pea proteins. Emulsions stabilised by these mixtures showed slightly different behaviour when produced at pH 7.0 or pH 3.8. © 2014 Society of Chemical Industry.

  15. Pea weevil, Bruchus pisorum (L.) (Coleoptera: Bruchidae), resistance in Pisum sativum x P. fulvum interspecific crosses

    USDA-ARS?s Scientific Manuscript database

    The pea weevil, Bruchus pisorum (L.), is one of the most intractable pest problems of cultivated pea, Pisum sativum L., in the world. This study investigated the transfer of pea weevil resistance from two accessions (PI 595946, PI 343955) of wild pea, Pisum fulvum Sibth. & Sm., to interspecific pop...

  16. Integrated effect of microbial antagonist, organic amendment and fungicide in controlling seedling mortality (Rhizoctonia solani) and improving yield in pea (Pisum sativum L.).

    PubMed

    Akhter, Wasira; Bhuiyan, Mohamed Khurshed Alam; Sultana, Farjana; Hossain, Mohamed Motaher

    2015-01-01

    The study evaluated the comparative performance of a few microbial antagonists, organic amendments and fungicides and their integration for the management of seedling mortality (Rhizoctonia solani Kühn) and yield improvement in pea (Pisum sativum L.). Before setting the experiment in field microplots, a series of in vitro and in vivo experiments were conducted to select a virulent isolate of R. solani, an effective antagonistic isolate of Trichoderma harzianum, a fungitoxic organic amendment and an appropriate fungicide. A greenhouse pathogenicity test compared differences in seedling mortality in pea inoculated by four isolates of R. solani and identified the isolate RS10 as the most virulent one. Among the 20 isolates screened in dual culture assay on PDA, T. harzianum isolate T-3 was found to show the highest (77.22%) inhibition of the radial growth of R. solani. A complete inhibition (100.00%) of colony growth of R. solani was observed when fungicide Bavistin 50 WP and Provax-200 at the rate of 100 and 250 ppm, respectively, were used, while Provax-200 was found to be highly compatible with T. harzianum. Mustard oilcake gave maximum inhibition (60.28%) of the radial growth of R. solani at all ratios, followed by sesame oilcake and tea waste. Integration of soil treatment with T. harzianum isolate T-3 and mustard oilcake and seed treatment with Provax-200 appeared to be significantly superior in reducing seedling mortality and improving seed yield in pea in comparison to any single or dual application of them in the experimental field. The research results will help growers develop integrated disease management strategies for the control of Rhizoctonia disease in pea. The research results show the need for an integrating selective microbial antagonist, organic amendment and fungicide to achieve appropriate management of seedling mortality (R. solani) and increase of seed yield in pea.

  17. Radiations from hot nuclei

    NASA Technical Reports Server (NTRS)

    Malik, F. Bary

    1993-01-01

    The investigation indicates that nuclei with excitation energy of a few hundred MeV to BeV are more likely to radiate hot nuclear clusters than neutrons. These daughter clusters could, furthermore, de-excite emitting other hot nuclei, and the chain continues until these nuclei cool off sufficiently to evaporate primarily neutrons. A few GeV excited nuclei could radiate elementary particles preferentially over neutrons. Impact of space radiation with materials (for example, spacecraft) produces highly excited nuclei which cool down emitting electromagnetic and particle radiations. At a few MeV excitation energy, neutron emission becomes more dominant than gamma-ray emission and one often attributes the cooling to take place by successive neutron decay. However, a recent experiment studying the cooling process of 396 MeV excited Hg-190 casts some doubt on this thinking, and the purpose of this investigation is to explore the possibility of other types of nuclear emission which might out-compete with neutron evaporation.

  18. Expression of the 12-oxophytodienoic acid 10,11-reductase gene in the compatible interaction between pea and fungal pathogen.

    PubMed

    Ishiga, Yasuhiro; Funato, Akiko; Tachiki, Tomoyuki; Toyoda, Kazuhiro; Shiraishi, Tomonori; Yamada, Tetsuji; Ichinose, Yuki

    2002-10-01

    Suppressors produced by Mycosphaerella pinodes are glycopeptides to block pea defense responses induced by elicitors. A clone, S64, was isolated as cDNA for suppressor-inducible gene from pea epicotyls. The treatment of pea epicotyls with suppressor alone induced an increase of S64 mRNA within 1 h, and it reached a maximum level at 3 h after treatment. The induction was not affected by application of the elicitor, indicating that the suppressor has a dominant action to regulate S64 gene expression. S64 was also induced by inoculation with a virulent pathogen, M. pinodes, but not by inoculation with a non-pathogen, Ascochyta rabiei, nor by treatment with fungal elicitor. The deduced structure of S64 showed high homology to 12-oxophytodienoic acid reductase (OPR) in Arabidopsis thaliana. A recombinant protein derived from S64 had OPR activity, suggesting compatibility-specific activation of the octadecanoid pathway in plants. Treatment with jasmonic acid (JA) or methyl jasmonic acid, end products of the octadecanoid pathway, inhibited the elicitor-induced accumulation of PAL mRNA in pea. These results indicate that the suppressor-induced S64 gene expression leads to the production of JA or related compounds, which might contribute to the establishment of compatibility by inhibiting the phenylpropanoid biosynthetic pathway.

  19. Differential gene expression in C4 plants. Research proposal, February 1, 1982-January 31, 1983. [Pea plants

    SciTech Connect

    Cashmore, A. R.

    1981-11-01

    The topic of this research proposal is slightly different from that originally outlined. Specifically, instead of characterizing the genes encoding the small subunit of RuBP carboxylase and the chlorophyll a/b binding polypeptide from corn, these genes from pea are being characterized. The above polypeptides represent the major products of cytoplasmic protein synthesis in green leaves. CDNA clones encoding the above polypeptides were isolated and characterized. Both of these cDNA clones have now been sequenced, providing the amino acid sequences for the carboxylase small subunit and, for the first time, for the chlorophyll a/b binding polypeptide. Pea nuclear DNA was cloned into the lambda phage Charon 4, and cloned nuclear DNA sequences encoding the above polypeptides were isolated. Future work will be concerned with the structural and functional characterization of these nuclear genes.

  20. An efficient technique for nuclei segmentation based on ellipse descriptor analysis and improved seed detection algorithm.

    PubMed

    Xu, Hongming; Lu, Cheng; Mandal, Mrinal

    2014-09-01

    In this paper, we propose an efficient method for segmenting cell nuclei in the skin histopathological images. The proposed technique consists of four modules. First, it separates the nuclei regions from the background with an adaptive threshold technique. Next, an elliptical descriptor is used to detect the isolated nuclei with elliptical shapes. This descriptor classifies the nuclei regions based on two ellipticity parameters. Nuclei clumps and nuclei with irregular shapes are then localized by an improved seed detection technique based on voting in the eroded nuclei regions. Finally, undivided nuclei regions are segmented by a marked watershed algorithm. Experimental results on 114 different image patches indicate that the proposed technique provides a superior performance in nuclei detection and segmentation.

  1. Search for Superheavy Nuclei

    NASA Astrophysics Data System (ADS)

    Hamilton, J. H.; Hofmann, S.; Oganessian, Y. T.

    2013-10-01

    We describe the discoveries of new superheavy nuclei (a) with Z=107-112 produced in cold fusion reactions between 208Pb and 209Bi and beams of A > 50 and (b) with Z=113-118 in hot fusion reactions between actinide nuclei and 48Ca. We also discuss the facilities used in these measurements. We compare the behavior of the β-decay energies and half-lives, spontaneous fission half-lives, cross sections, and excitation functions with expectations from theoretical calculations. Finally, we outline future research directions, including studies of the detailed properties of nuclei synthesized at higher yields, searches for new elements with Z=119 and 120, and developments of new facilities.

  2. Scattering Of Light Nuclei

    SciTech Connect

    Quaglioni, S; Navratil, P; Roth, R

    2009-12-15

    The exact treatment of nuclei starting from the constituent nucleons and the fundamental interactions among them has been a long-standing goal in nuclear physics. Above all nuclear scattering and reactions, which require the solution of the many-body quantum-mechanical problem in the continuum, represent an extraordinary theoretical as well as computational challenge for ab initio approaches.We present a new ab initio many-body approach which derives from the combination of the ab initio no-core shell model with the resonating-group method [4]. By complementing a microscopic cluster technique with the use of realistic interactions, and a microscopic and consistent description of the nucleon clusters, this approach is capable of describing simultaneously both bound and scattering states in light nuclei. We will discuss applications to neutron and proton scattering on sand light p-shell nuclei using realistic nucleon-nucleon potentials, and outline the progress toward the treatment of more complex reactions.

  3. Characterization and pathogenicity of Rhizoctonia and Rhizoctonia-like spp. from pea crops in the Columbia Basin of Oregon and Washington

    USDA-ARS?s Scientific Manuscript database

    A total of 179 isolates of Rhizoctonia and Rhizoctonia-like species were obtained from soil and plant samples collected from irrigated pea crops in the semi-arid Columbia Basin of Oregon and Washington from 2011 to 2013, and characterized to species, subspecies, and anastomosis groups (AG) based on ...

  4. The shapes of nuclei

    NASA Astrophysics Data System (ADS)

    Bertsch, G. F.

    Gerry Brown initiated some early studies on the coexistence of different nuclear shapes. The subject has continued to be of interest and is crucial for understanding nuclear fission. We now have a very good picture of the potential energy surface with respect to shape degrees of freedom in heavy nuclei, but the dynamics remain problematic. In contrast, the early studies on light nuclei were quite successful in describing the mixing between shapes. Perhaps a new approach in the spirit of the old calculations could better elucidate the character of the fission dynamics and explain phenomena that current theory does not model well.

  5. Disintegration of comet nuclei

    NASA Astrophysics Data System (ADS)

    Ksanfomality, Leonid V.

    2012-02-01

    The breaking up of comets into separate pieces, each with its own tail, was seen many times by astronomers of the past. The phenomenon was in sharp contrast to the idea of the eternal and unchangeable celestial firmament and was commonly believed to be an omen of impending disaster, especially for comets with tails stretching across half the sky. It is only now that we have efficient enough space exploration tools to see comet nuclei and even - in the particular case of small comet Hartley-2 in 2010 - to watch their disintegration stage. There are also other suspected candidates for disintegration in the vast family of comet nuclei and other Solar System bodies.

  6. Reaction theory for exotic nuclei

    SciTech Connect

    Bonaccorso, Angela

    2014-05-09

    Exotic nuclei are usually defined as those with unusual N/Z ratios. They can be found in the crust of neutron stars enbedded in a sea of electrons or created in laboratory by fragmentation of a primary beam (in-flight method) or of the target (ISOL method). They are extremely important for nuclear astrophysics, see for example Ref.[1]. Furthermore by studying them we can check the limits of validity of nuclear reaction and structure models. This contribution will be devoted to the understanding of how by using reaction theory and comparing to the data we can extract structure information. We shall discuss the differences between the mechanisms of transfer and breakup reactions, an we will try to explain how nowadays it is possible to do accurate spectroscopy in extreme conditions.

  7. Neutral Peptidases in the Stroma of Pea Chloroplasts 1

    PubMed Central

    Liu, Xiang-Qin; Jagendorf, André T.

    1986-01-01

    One endopeptidase (EP1) and at least three aminopeptidases (AP1, AP2, and AP3) were discovered in the stroma of chloroplasts isolated from pea seedlings (Pisum sativum L.), and purified over 100-fold. EP1 requires added Mg2+ or Ca2+ for activity, may have an additional tightly bound metal atom, and is inhibited by sulfhydryl reagents but not by serine residue-directed inhibitors. It is reversibly inhibited by dithiothreitol. Its specificity is for the bond between two adjacent Ala or Gly residues. Its molecular mass is 93 kilodaltons, estimated on a gel filtration column. Aminopeptidase activities were detected with the aid of different amino acyl-β-naphthylamides as substrates. They were resolved into at least three individual proteins by gel filtration and DEAE-cellulose chromatography, having apparent molecular masses of 269,000 (AP1), 84,000 (AP2), and 42,000 (AP3) daltons, respectively. Each has a unique specificity for substrates, with AP1 hydrolyzing only the Prolyl-β-naphthylamide. None of the APs require added divalent cations for activity, but the possibility of a tightly bound metal function was suggested in AP2 and AP3 (not AP1) from effects of inhibitors. A probable sulfhydryl residue function was indicated for all three, from inhibition by p-hydroxymercuribenzoate and Zn2+. All these peptidases had pH optima at 7.7. PMID:16664864

  8. Physics with Polarized Nuclei.

    ERIC Educational Resources Information Center

    Thompson, William J.; Clegg, Thomas B.

    1979-01-01

    Discusses recent advances in polarization techniques, specifically those dealing with polarization of atomic nuclei, and how polarized beams and targets are produced. These techniques have greatly increased the scope of possible studies, and provided the tools for testing fundamental symmetries and the spin dependence of nuclear forces. (GA)

  9. The effect of microgravity on proton permeability of thylakoid membranes and contribution of II and I photosystems in photosynthetic electron transport in pea chloroplasts.

    PubMed

    Zolotareva, E K; Onoiko, E B; Sytnik, S K; Podorvanov, V V

    1999-07-01

    According to a number investigations microgravity conditions affect membrane apparatus of photosynthesis in cells of higher plants and alga [for review, see Kordyum et al., 1994; Kordyum, 1997]. (see for review). Chloroplasts of space-grown pea plants showed disintegration of grana, shrinkage of the membrane constituting the grana stacks and other structural perturbance of the photosynthetic membranes. However there have been no studies on the effect of microgravity on proton permeability of thylakoid membranes and closely connected with this parameter their photochemical characteristics. The aim of the study is investigation of microgravity effects on protonic permeability of photosynthetic membrane and contribution of photosystem II (PSII) and photosystem I (PSI) in electron transfer from water to potassium ferrycianide (FeCy) in isolated pea chloroplasts. Pea.

  10. Cloning and Characterization of TPE4A, a Thiol-Protease Gene Induced during Ovary Senescence and Seed Germination in Pea1

    PubMed Central

    Cercós, Manuel; Santamaría, Salvador; Carbonell, Juan

    1999-01-01

    A cDNA clone encoding a thiol-protease (TPE4A) was isolated from senescent ovaries of pea (Pisum sativum) by reverse transcriptase-polymerase chain reaction. The deduced amino acid sequence of TPE4A has the conserved catalytic amino acids of papain. It is very similar to VSCYSPROA, a thiol-protease induced during seed germination in common vetch. TPE4A mRNA levels increase during the senescence of unpollinated pea ovaries and are totally suppressed by treatment with gibberellic acid. In situ hybridization indicated that TPE4A mRNA distribution in senescent pea ovaries is different from that of previously reported thiol-proteases induced during senescence, suggesting the involvement of different proteases in the mobilization of proteins from senescent pea ovaries. TPE4A is also induced during the germination of pea seeds, indicating that a single protease gene can be induced during two different physiological processes, senescence and germination, both of which require protein mobilization. PMID:10198093

  11. The Pea GIGAS Gene Is a FLOWERING LOCUS T Homolog Necessary for Graft-Transmissible Specification of Flowering but Not for Responsiveness to Photoperiod[C][W

    PubMed Central

    Hecht, Valérie; Laurie, Rebecca E.; Vander Schoor, Jacqueline K.; Ridge, Stephen; Knowles, Claire L.; Liew, Lim Chee; Sussmilch, Frances C.; Murfet, Ian C.; Macknight, Richard C.; Weller, James L.

    2011-01-01

    Garden pea (Pisum sativum) was prominent in early studies investigating the genetic control of flowering and the role of mobile flowering signals. In view of recent evidence that genes in the FLOWERING LOCUS T (FT) family play an important role in generating mobile flowering signals, we isolated the FT gene family in pea and examined the regulation and function of its members. Comparison with Medicago truncatula and soybean (Glycine max) provides evidence of three ancient subclades (FTa, FTb, and FTc) likely to be common to most crop and model legumes. Pea FT genes show distinctly different expression patterns with respect to developmental timing, tissue specificity, and response to photoperiod and differ in their activity in transgenic Arabidopsis thaliana, suggesting they may have different functions. We show that the pea FTa1 gene corresponds to the GIGAS locus, which is essential for flowering under long-day conditions and promotes flowering under short-day conditions but is not required for photoperiod responsiveness. Grafting, expression, and double mutant analyses show that GIGAS/FTa1 regulates a mobile flowering stimulus but also provide clear evidence for a second mobile flowering stimulus that is correlated with expression of FTb2 in leaf tissue. These results suggest that induction of flowering by photoperiod in pea results from interactions among several members of a diversified FT family. PMID:21282524

  12. Lyα Profile, Dust, and Prediction of Lyα Escape Fraction in Green Pea Galaxies

    NASA Astrophysics Data System (ADS)

    Yang, Huan; Malhotra, Sangeeta; Gronke, Max; Rhoads, James E.; Leitherer, Claus; Wofford, Aida; Jiang, Tianxing; Dijkstra, Mark; Tilvi, V.; Wang, Junxian

    2017-08-01

    We studied Lyman-α (Lyα) escape in a statistical sample of 43 Green Peas with HST/COS Lyα spectra. Green Peas are nearby star-forming galaxies with strong [O iii]λ5007 emission lines. Our sample is four times larger than the previous sample and covers a much more complete range of Green Pea properties. We found that about two-thirds of Green Peas are strong Lyα line emitters with rest-frame Lyα equivalent width > 20 \\mathringA . The Lyα profiles of Green Peas are diverse. The Lyα escape fraction, defined as the ratio of observed Lyα flux to intrinsic Lyα flux, shows anti-correlations with a few Lyα kinematic features—both the blue peak and red peak velocities, the peak separations, and the FWHM of the red portion of the Lyα profile. Using properties measured from Sloan Digital Sky Survey optical spectra, we found many correlations—the Lyα escape fraction generally increases at lower dust reddening, lower metallicity, lower stellar mass, and higher [O iii]/[O ii] ratio. We fit their Lyα profiles with the H i shell radiative transfer model and found that the Lyα escape fraction is anti-correlated with the best-fit N H i . Finally, we fit an empirical linear relation to predict {f}{esc}{Lyα } from the dust extinction and Lyα red peak velocity. The standard deviation of this relation is about 0.3 dex. This relation can be used to isolate the effect of intergalactic medium (IGM) scatterings from Lyα escape and to probe the IGM optical depth along the line of sight of each z> 7 Lyα emission-line galaxy in the James Webb Space Telescope era.

  13. [Fasciation in pea: basic principles of morphogenesis].

    PubMed

    Siniushin, A A; Gostimskiĭ, S A

    2006-01-01

    A study of fasciated pea Pisum sativum L. (Fabaceae) mutant Shtambovy in comparison with the wild type (Nemchinovsky cultivar) has shown that fasciation is a result of abnormal cohesion of axial or other structures which arise in a superfluous amount due to uncontrolled meristic processes. In some cases, the organs with the same number and position as in the wild type can be fascinated. Subsequent defasciation and some features of tissue differentiation suggest that the meristem of a fasciated shoot retains a certain degree of discreteness which reflects its complex structure. The number and position of leaves in a node is a function of the diameter of the leaf primordium inhibitory zone, size of the shoot apical meristem, and number of bundles in a shoot. In the absence of the apex proliferative activity combined with the reduction of phyllomes in the upper nodes, abnormal cohesion of the second order axes, racemes, can take place. As a result, inflorescences of special type develop.

  14. Protein methylation reactions in intact pea chloroplasts

    SciTech Connect

    Niemi, K.J. )

    1989-04-01

    Post-translational protein methylation was investigated in Pisum sativum chloroplasts. Intact pea chloroplasts were incubated with ({sup 3}H-methyl)-S-adenosylmethionine under various conditions. The chloroplasts were then separated into stromal and thylakoid fractions and analyzed for radioactivity transferred to protein. Light enhanced the magnitude of labeling in both fractions. One thylakoid polypeptide with an apparent molecular mass of 43 kDa was labeled only in the light. Several other thylakoid and stromal proteins were labeled in both light and dark-labeling conditions. Both base-labile methylation, carboxy-methylesters and base-stable groups, N-methylations were found. Further characterization of the methyl-transfer reactions will be presented.

  15. Ozone stress proteins in pea plants

    SciTech Connect

    Beckett, P.; Mozley, D.; Price, A.; Hetherington, A.; Lea, P. )

    1989-04-01

    21 day old pea plants were fumigated with 200, 100, 50 and 0 ppb ozone (8 hrs/day) for 5 days. Soluble proteins were extracted from the first 6 leaves and analyzed by 1D SDS PAGE. Polypeptides were visualized after coomassie blue staining. With respect to controls, fumigation resulted in a dose dependent decrease in staining intensity of several polypeptides (of approximate M.W. 94, 54, 35 kD). However, treatment with 200 ppb ozone resulted in the appearance of a polypeptide with a molecular weight of circa 32 kD. This polypeptide was absent from control (0 ppb ozone) plants. Currently, we are (1) purifying the c32kD polypeptide, (2) studying the temporal aspects of the synthesis of this polypeptide and (3) investigating whether this represents a general response to pollutant gases.

  16. Analysis of the accumulation of Pea enation mosaic virus genomes in seed tissues and lack of evidence for seed transmission in pea (Pisum sativum L.)

    USDA-ARS?s Scientific Manuscript database

    Pea enation mosaic virus (PEMV) is an important virus disease of pea. International movement of commercial pea cultivars and germplasm can be problematic due to uncertainty about seed transmission of the viruses responsible for the disease. Whether PEMV is seed-borne was assessed by collecting dev...

  17. 7 CFR 457.137 - Green pea crop insurance provisions.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... CORPORATION, DEPARTMENT OF AGRICULTURE COMMON CROP INSURANCE REGULATIONS § 457.137 Green pea crop insurance... operation of harvesting equipment; and (ii) Abnormally hot or cold temperatures that cause an unexpected...

  18. 7 CFR 457.137 - Green pea crop insurance provisions.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... CORPORATION, DEPARTMENT OF AGRICULTURE COMMON CROP INSURANCE REGULATIONS § 457.137 Green pea crop insurance... harvesting equipment; and (ii) Abnormally hot or cold temperatures that cause an unexpected number of acres...

  19. Lyα and UV Sizes of Green Pea Galaxies

    NASA Astrophysics Data System (ADS)

    Yang, Huan; Malhotra, Sangeeta; Rhoads, James E.; Leitherer, Claus; Wofford, Aida; Jiang, Tianxing; Wang, Junxian

    2017-03-01

    Green Peas are nearby analogs of high-redshift Lyα-emitting galaxies (LAEs). To probe their Lyα escape, we study the spatial profiles of Lyα and UV continuum emission of 24 Green Pea galaxies using the Cosmic Origins Spectrograph (COS) on the Hubble Space Telescope. We extract the spatial profiles of Lyα emission from their 2D COS spectra, and of the UV continuum from both 2D spectra and NUV images. The Lyα emission shows more extended spatial profiles than the UV continuum, in most Green Peas. The deconvolved full width at half maximum of the Lyα spatial profile is about 2-4 times that of the UV continuum, in most cases. Because Green Peas are analogs of high z LAEs, our results suggest that most high-z LAEs probably have larger Lyα sizes than UV sizes. We also compare the spatial profiles of Lyα photons at blueshifted and redshifted velocities in eight Green Peas with sufficient data quality, and find that the blue wing of the Lyα line has a larger spatial extent than the red wing in four Green Peas with comparatively weak blue Lyα line wings. We show that Green Peas and MUSE z = 3-6 LAEs have similar Lyα and UV continuum sizes, which probably suggests that starbursts in both low-z and high-z LAEs drive similar gas outflows illuminated by Lyα light. Five Lyman continuum (LyC) leakers in this sample have similar Lyα to UV continuum size ratios (˜1.4-4.3) to the other Green Peas, indicating that their LyC emissions escape through ionized holes in the interstellar medium.

  20. RNA-sequencing from single nuclei.

    PubMed

    Grindberg, Rashel V; Yee-Greenbaum, Joyclyn L; McConnell, Michael J; Novotny, Mark; O'Shaughnessy, Andy L; Lambert, Georgina M; Araúzo-Bravo, Marcos J; Lee, Jun; Fishman, Max; Robbins, Gillian E; Lin, Xiaoying; Venepally, Pratap; Badger, Jonathan H; Galbraith, David W; Gage, Fred H; Lasken, Roger S

    2013-12-03

    It has recently been established that synthesis of double-stranded cDNA can be done from a single cell for use in DNA sequencing. Global gene expression can be quantified from the number of reads mapping to each gene, and mutations and mRNA splicing variants determined from the sequence reads. Here we demonstrate that this method of transcriptomic analysis can be done using the extremely low levels of mRNA in a single nucleus, isolated from a mouse neural progenitor cell line and from dissected hippocampal tissue. This method is characterized by excellent coverage and technical reproducibility. On average, more than 16,000 of the 24,057 mouse protein-coding genes were detected from single nuclei, and the amount of gene-expression variation was similar when measured between single nuclei and single cells. Several major advantages of the method exist: first, nuclei, compared with whole cells, have the advantage of being easily isolated from complex tissues and organs, such as those in the CNS. Second, the method can be widely applied to eukaryotic species, including those of different kingdoms. The method also provides insight into regulatory mechanisms specific to the nucleus. Finally, the method enables dissection of regulatory events at the single-cell level; pooling of 10 nuclei or 10 cells obscures some of the variability measured in transcript levels, implying that single nuclei and cells will be extremely useful in revealing the physiological state and interconnectedness of gene regulation in a manner that avoids the masking inherent to conventional transcriptomics using bulk cells or tissues.

  1. RNA-sequencing from single nuclei

    PubMed Central

    Grindberg, Rashel V.; Yee-Greenbaum, Joyclyn L.; McConnell, Michael J.; Novotny, Mark; O’Shaughnessy, Andy L.; Lambert, Georgina M.; Araúzo-Bravo, Marcos J.; Lee, Jun; Fishman, Max; Robbins, Gillian E.; Lin, Xiaoying; Venepally, Pratap; Badger, Jonathan H.; Galbraith, David W.; Gage, Fred H.; Lasken, Roger S.

    2013-01-01

    It has recently been established that synthesis of double-stranded cDNA can be done from a single cell for use in DNA sequencing. Global gene expression can be quantified from the number of reads mapping to each gene, and mutations and mRNA splicing variants determined from the sequence reads. Here we demonstrate that this method of transcriptomic analysis can be done using the extremely low levels of mRNA in a single nucleus, isolated from a mouse neural progenitor cell line and from dissected hippocampal tissue. This method is characterized by excellent coverage and technical reproducibility. On average, more than 16,000 of the 24,057 mouse protein-coding genes were detected from single nuclei, and the amount of gene-expression variation was similar when measured between single nuclei and single cells. Several major advantages of the method exist: first, nuclei, compared with whole cells, have the advantage of being easily isolated from complex tissues and organs, such as those in the CNS. Second, the method can be widely applied to eukaryotic species, including those of different kingdoms. The method also provides insight into regulatory mechanisms specific to the nucleus. Finally, the method enables dissection of regulatory events at the single-cell level; pooling of 10 nuclei or 10 cells obscures some of the variability measured in transcript levels, implying that single nuclei and cells will be extremely useful in revealing the physiological state and interconnectedness of gene regulation in a manner that avoids the masking inherent to conventional transcriptomics using bulk cells or tissues. PMID:24248345

  2. Energetic Nuclei, Superdensity and Biomedicine

    ERIC Educational Resources Information Center

    Baldin, A. M.

    1977-01-01

    High-energy, relativistic nuclei were first observed in cosmic rays. Studing these nuclei has provided an opportunity for analyzing the composition of cosmic rays and for experimentally verifying principles governing the behavior of nuclear matter at high and super-high temperatures. Medical research using accelerated nuclei is suggested.…

  3. Energetic Nuclei, Superdensity and Biomedicine

    ERIC Educational Resources Information Center

    Baldin, A. M.

    1977-01-01

    High-energy, relativistic nuclei were first observed in cosmic rays. Studing these nuclei has provided an opportunity for analyzing the composition of cosmic rays and for experimentally verifying principles governing the behavior of nuclear matter at high and super-high temperatures. Medical research using accelerated nuclei is suggested.…

  4. Exotic phenomena in nuclei

    NASA Astrophysics Data System (ADS)

    Neff, Thomas; Feldmeier, Hans; Roth, Robert

    2006-10-01

    In the Fermionic Molecular Dynamics (FMD) model the nuclear many-body system is described using Slater determinants with Gaussian wave-packets as single-particle states. The flexibility of the FMD wave functions allows for a consistent description of shell model like structures, deformed states, cluster structures as well as halos. An effective interaction derived from the realistic Argonne V18 interaction using the Unitary Correlation Operator Method is used for all nuclei. Results for nuclei in the p-shell will be presented. Halo features are present in the Helium isotopes, cluster structures are studied in Beryllium and Carbon isotopes. The interplay between shell structure and cluster structures in the ground and the Hoyle state in ^12C will be discussed.

  5. Effect of cationic plastoquinone SkQ1 on electron transfer reactions in chloroplasts and mitochondria from pea seedlings.

    PubMed

    Samuilov, V D; Kiselevsky, D B

    2015-04-01

    Plastoquinone bound with decyltriphenylphosphonium cation (SkQ1) penetrating through the membrane in nanomolar concentrations inhibited H2O2 generation in cells of epidermis of pea seedling leaves that was detected by the fluorescence of 2',7'-dichlorofluorescein. Photosynthetic electron transfer in chloroplasts isolated from pea leaves is suppressed by SkQ1 at micromolar concentrations: the electron transfer in chloroplasts under the action of photosystem II or I (with silicomolybdate or methyl viologen as electron acceptors, respectively) is more sensitive to SkQ1 than under the action of photosystem II + I (with ferricyanide or p-benzoquinone as electron acceptors). SkQ1 reduced by borohydride is oxidized by ferricyanide, p-benzoquinone, and, to a lesser extent, by silicomolybdate, but not by methyl viologen. SkQ1 is not effective as an electron acceptor supporting O2 evolution from water in illuminated chloroplasts. The data on suppression of photosynthetic O2 evolution or consumption show that SkQ1, similarly to phenazine methosulfate, causes conversion of the chloroplast redox-chain from non-cyclic electron transfer mode to the cyclic mode without O2 evolution. Oxidation of NADH or succinate in mitochondria isolated from pea roots is stimulated by SkQ1.

  6. Heat-Induced Soluble Protein Aggregates from Mixed Pea Globulins and β-Lactoglobulin.

    PubMed

    Chihi, Mohamed-Lazhar; Mession, Jean-luc; Sok, Nicolas; Saurel, Rémi

    2016-04-06

    The present work investigates the formation of protein aggregates (85 °C, 60 min incubation) upon heat treatment of β-lactoglobulin (βlg)-pea globulins (Glob) mixtures at pH 7.2 and 5 mM NaCl from laboratory-prepared protein isolates. Various βlg/Glob weight ratios were applied, for a total protein concentration of 2 wt % in admixture. Different analytical methods were used to determine the aggregation behavior of "mixed" aggregates, that is, surface hydrophobicity and also sulfhydryl content, protein interactions by means of SDS-PAGE electrophoresis, and molecule size distribution by DLS and gel filtration. The production of "mixed" thermal aggregates would involve both the formation of new disulfide bonds and noncovalent interactions between the denatured βlg and Glob subunits. The majority of "mixed" soluble aggregates displayed higher molecular weight and smaller diameter than those for Glob heated in isolation. The development of pea-whey protein "mixed" aggregates may help to design new ingredients for the control of innovative food textures.

  7. Nucleotide sequence of a complementary DNA encoding pea cytosolic copper/zinc superoxide dismutase. [Pisum sativum L

    SciTech Connect

    White, D.A.; Zilinskas, B.A. )

    1991-08-01

    The authors now report the nucleotide sequence of the cytosolic Cu/Zn SOD cloned from a {lambda}gt11 cDNA library constructed from mRNA extracted from leaves of 7- to 10-d pea seedlings (Pisum sativum L.). The clone was isolated using a 22-base synthetic oligonucleotide complementary to the amino acid sequence CGIIGLQG. This sequence, found at the protein's carboxy terminus, is highly conserved among plant cytosolic Cu/Zn SODs but not chloroplastic Cu/Zn SODs. The 738-base pair sequence contains an open reading frame specifying 152 codons and a predicted M{sub r} of 18,024 D. The deduced amino acid sequence is highly homologous (79-82% identity) with the sequences of other known plant cytosolic Cu/Zn SODs but less highly conserved (63-65%) when compared with several chloroplastic Cu/Zn SODs including pea (10).

  8. Protein kinase B/Akt binds and phosphorylates PED/PEA-15, stabilizing its antiapoptotic action.

    PubMed

    Trencia, Alessandra; Perfetti, Anna; Cassese, Angela; Vigliotta, Giovanni; Miele, Claudia; Oriente, Francesco; Santopietro, Stefania; Giacco, Ferdinando; Condorelli, Gerolama; Formisano, Pietro; Beguinot, Francesco

    2003-07-01

    The antiapoptotic protein PED/PEA-15 features an Akt phosphorylation motif upstream from Ser(116). In vitro, recombinant PED/PEA-15 was phosphorylated by Akt with a stoichiometry close to 1. Based on Western blotting with specific phospho-Ser(116) PED/PEA-15 antibodies, Akt phosphorylation of PED/PEA-15 occurred mainly at Ser(116). In addition, a mutant of PED/PEA-15 featuring the substitution of Ser(116)-->Gly (PED(S116-->G)) showed 10-fold-decreased phosphorylation by Akt. In intact 293 cells, Akt also induced phosphorylation of PED/PEA-15 at Ser(116). Based on pull-down and coprecipitation assays, PED/PEA-15 specifically bound Akt, independently of Akt activity. Serum activation of Akt as well as BAD phosphorylation by Akt showed no difference in 293 cells transfected with PED/PEA-15 and in untransfected cells (which express no endogenous PED/PEA-15). However, the antiapoptotic action of PED/PEA-15 was almost twofold reduced in PED(S116-->G) compared to that in PED/PEA-15(WT) cells. PED/PEA-15 stability closely paralleled Akt activation by serum in 293 cells. In these cells, the nonphosphorylatable PED(S116-->G) mutant exhibited a degradation rate threefold greater than that observed with wild-type PED/PEA-15. In the U373MG glioma cells, blocking Akt also reduced PED/PEA-15 levels and induced sensitivity to tumor necrosis factor-related apoptosis-inducing ligand apoptosis. Thus, phosphorylation by Akt regulates the antiapoptotic function of PED/PEA-15 at least in part by controlling the stability of PED/PEA-15. In part, Akt survival signaling may be mediated by PED/PEA-15.

  9. Purification and Characterization of Pea Epicotyl beta-Amylase.

    PubMed

    Lizotte, P A; Henson, C A; Duke, S H

    1990-03-01

    The most abundant beta-amylase (EC 3.2.1.2) in pea (Pisum sativum L.) was purified greater than 880-fold from epicotyls of etiolated germinating seedlings by anion exchange and gel filtration chromatography, glycogen precipitation, and preparative electrophoresis. The electrophoretic mobility and relative abundance of this beta-amylase are the same as that of an exoamylase previously reported to be primarily vacuolar. The enzyme was determined to be a beta-amylase by end product analysis and by its inability to hydrolyze beta-limit dextrin and to release dye from starch azure. Pea beta-amylase is an approximate 55 to 57 kilodalton monomer with a pl of 4.35, a pH optimum of 6.0 (soluble starch substrate), an Arrhenius energy of activation of 6.28 kilocalories per mole, and a K(m) of 1.67 milligrams per milliliter (soluble starch). The enzyme is strongly inhibited by heavy metals, p-chloromer-curiphenylsulfonic acid and N-ethylmaleimide, but much less strongly by iodoacetamide and iodoacetic acid, indicating cysteinyl sulfhydryls are not directly involved in catalysis. Pea beta-amylase is competitively inhibited by its end product, maltose, with a K(i) of 11.5 millimolar. The enzyme is partially inhibited by Schardinger maltodextrins, with alpha-cyclohexaamylose being a stronger inhibitor than beta-cycloheptaamylose. Moderately branched glucans (e.g. amylopectin) were better substrates for pea beta-amylase than less branched or non-branched (amyloses) or highly branched (glycogens) glucans. The enzyme failed to hydrolyze native starch grains from pea and glucans smaller than maltotetraose. The mechanism of pea beta-amylase is the multichain type. Possible roles of pea beta-amylase in cellular glucan metabolism are discussed.

  10. Gene-based SNP discovery and genetic mapping in pea.

    PubMed

    Sindhu, Anoop; Ramsay, Larissa; Sanderson, Lacey-Anne; Stonehouse, Robert; Li, Rong; Condie, Janet; Shunmugam, Arun S K; Liu, Yong; Jha, Ambuj B; Diapari, Marwan; Burstin, Judith; Aubert, Gregoire; Tar'an, Bunyamin; Bett, Kirstin E; Warkentin, Thomas D; Sharpe, Andrew G

    2014-10-01

    Gene-based SNPs were identified and mapped in pea using five recombinant inbred line populations segregating for traits of agronomic importance. Pea (Pisum sativum L.) is one of the world's oldest domesticated crops and has been a model system in plant biology and genetics since the work of Gregor Mendel. Pea is the second most widely grown pulse crop in the world following common bean. The importance of pea as a food crop is growing due to its combination of moderate protein concentration, slowly digestible starch, high dietary fiber concentration, and its richness in micronutrients; however, pea has lagged behind other major crops in harnessing recent advances in molecular biology, genomics and bioinformatics, partly due to its large genome size with a large proportion of repetitive sequence, and to the relatively limited investment in research in this crop globally. The objective of this research was the development of a genome-wide transcriptome-based pea single-nucleotide polymorphism (SNP) marker platform using next-generation sequencing technology. A total of 1,536 polymorphic SNP loci selected from over 20,000 non-redundant SNPs identified using deep transcriptome sequencing of eight diverse Pisum accessions were used for genotyping in five RIL populations using an Illumina GoldenGate assay. The first high-density pea SNP map defining all seven linkage groups was generated by integrating with previously published anchor markers. Syntenic relationships of this map with the model legume Medicago truncatula and lentil (Lens culinaris Medik.) maps were established. The genic SNP map establishes a foundation for future molecular breeding efforts by enabling both the identification and tracking of introgression of genomic regions harbouring QTLs related to agronomic and seed quality traits.

  11. Dosage compensation and sex-specific epigenetic landscape of the X chromosome in the pea aphid.

    PubMed

    Richard, Gautier; Legeai, Fabrice; Prunier-Leterme, Nathalie; Bretaudeau, Anthony; Tagu, Denis; Jaquiéry, Julie; Le Trionnaire, Gaël

    2017-01-01

    Heterogametic species display a differential number of sex chromosomes resulting in imbalanced transcription levels for these chromosomes between males and females. To correct this disequilibrium, dosage compensation mechanisms involving gene expression and chromatin accessibility regulations have emerged throughout evolution. In insects, these mechanisms have been extensively characterized only in Drosophila but not in insects of agronomical importance. Aphids are indeed major pests of a wide range of crops. Their remarkable ability to switch from asexual to sexual reproduction during their life cycle largely explains the economic losses they can cause. As heterogametic insects, male aphids are X0, while females (asexual and sexual) are XX. Here, we analyzed transcriptomic and open chromatin data obtained from whole male and female individuals to evaluate the putative existence of a dosage compensation mechanism involving differential chromatin accessibility of the pea aphid's X chromosome. Transcriptomic analyses first showed X/AA and XX/AA expression ratios for expressed genes close to 1 in males and females, respectively, suggesting dosage compensation in the pea aphid. Analyses of open chromatin data obtained by Formaldehyde-Assisted Isolation of Regulatory Elements (FAIRE-seq) revealed a X chromosome chromatin accessibility globally and significantly higher in males than in females, while autosomes' chromatin accessibility is similar between sexes. Moreover, chromatin environment of X-linked genes displaying similar expression levels in males and females-and thus likely to be compensated-is significantly more accessible in males. Our results suggest the existence of an underlying epigenetic mechanism enhancing the X chromosome chromatin accessibility in males to allow X-linked gene dose correction between sexes in the pea aphid, similar to Drosophila. Our study gives new evidence into the comprehension of dosage compensation in link with chromatin biology in

  12. Pea enation mosaic virus genoma RNA contains no polyadenylate sequences and cannot be aminoacylated.

    PubMed

    German, T L; De Zoeten, G A; Hall, T C

    1978-01-01

    An active synthetase enzyme preparation from peas (Pisum sativum L.) did not catalyze the aminoacylation of pea enation mosaic virus RNA. The viral RNA was shown not to contain polyadenylic acid sequences.

  13. Olfactory cues from different plant species in host selection by female pea moths.

    PubMed

    Thöming, Gunda; Norli, Hans Ragnar

    2015-03-04

    In herbivorous insects specialized on few plant species, attraction to host odor may be mediated by volatiles common to all host species, by specific compounds, or combinations of both. The pea moth Cydia nigricana is an important pest of the pea. Volatile signatures of four host plant species were studied to identify compounds involved in pea moth host selection and to improve previously reported attractive volatile blends. P. sativum and alternative Fabaceae host species were compared regarding female attraction, oviposition, and larval performance. Pea moth females were strongly attracted to the sweet pea Lathyrus odoratus, but larval performance on that species was moderate. Chemical analyses of sweet pea odor and electrophysiological responses of moth antennae led to identification of seven sweet-pea-specific compounds and ten compounds common to all tested host species. Blends of these specific and common cues were highly attractive to mated pea moth females in wind tunnel and field experiments.

  14. Properties of Cometary Nuclei

    NASA Technical Reports Server (NTRS)

    Rahe, J.; Vanysek, V.; Weissman, P. R.

    1994-01-01

    Active long- and short-period comets contribute about 20 to 30 % of the major impactors on the Earth. Cometary nuclei are irregular bodies, typically a few to ten kilometers in diameter, with masses in the range 10(sup 15) to 10(sup 18) g. The nuclei are composed of an intimate mixture of volatile ices, mostly water ice and hydrocarbon and silicate grains. The composition is the closest to solar composition of any known bodies in the solar system. The nuclei appear to be weakly bonded agglomerations of smaller icy planetesimals, and material strengths estimated from observed tidal disruption events are fairly low, typically 10(sup 2) to 10(sup 4) N m(sup -2). Density estimates range between 0.2 and 1.2 g cm(sup -3) but are very poorly determined, if at all. As comets age they develop nonvolitile crusts on their surfaces which eventually render them inactive, similar in appearance to carbonaceous asteroids. However, dormant comets may continue to show sporadic activity and outbursts for some time before they become truly extinct. The source of the long-period comets is the Oort cloud, a vast spherical cloud of perhaps 10(sup 12) to 10(sup 13) comets surrounding the solar system and extending to interstellar distances. The likely source for short-period comets is the Kuiper belt. a ring of perhaps 10(sup 8) to 10(sup 10) remnant icy planetesimals beyond the orbit of Neptune, though some short-period comets may also be long-period comets from the Oort cloud which have been perturbed into short-period orbits.

  15. Skyrmions and Nuclei

    NASA Astrophysics Data System (ADS)

    Battye, R. A.; Manton, N. S.; Sutcliffe, P. M.

    We review recent work on the modelling of atomic nuclei as quantised Skyrmions, using Skyrme's original model with pion fields only. Skyrmions are topological soliton solutions, whose conserved topological charge B is identified with the baryon number of a nucleus. Apart from an energy and length scale, the Skyrme model has just one dimensionless parameter m, proportional to the pion mass. It has been found that a good fit to experimental nuclear data requires m to be of order 1. The Skyrmions for B up to 7 have been known for some time, and are qualitatively insensitive to whether m is zero or of order 1. However, for baryon numbers B = 8 and above, the Skyrmions have quite a compact structure for m of order 1, rather than the hollow polyhedral structure found when m = 0. One finds for baryon numbers which are multiples of four, that the Skyrmions are composed of B = 4 sub-units, as in the α-particle model of nuclei. The rational map ansatz gives a useful approximation to the Skyrmion solutions for all baryon numbers when m = 0. For m of order 1, it gives a good approximation for baryon numbers up to 7, and generalisations of this ansatz are helpful for higher baryon numbers. We briefly review the work from the 1980s and 90s on the semiclassical rigidbody quantisation of Skyrmions for B = 1, 2, 3 and 4. We then discuss more recent work extending this method to B = 6, 7, 8, 10 and 12. We determine the quantum states of the Skyrmions, finding their spins, isospins and parities, and compare with the experimental data on the ground and excited states of nuclei up to mass number 12.

  16. 3-Methyleneoxindole Reductase of Peas 1

    PubMed Central

    Moyed, H. S.; Williamson, Valerie

    1967-01-01

    A 100-fold purification of a reduced triphosphopyridine nucleotide/3-methyleneoxindole reductase of peas has been achieved using conventional protein fractionation procedures. Reduced diphosphopyridine nucleotide is 25-fold less effective than reduced triphosphopyridine nucleotide as the reductant. The preparation is free of other reductase activities including those linking the oxidation of reduced pyridine nucleotide coenzymes to the reduction of cytochrome c; vitamins K1, K2, and K3; O2; nitrate; oxidized glutathione; and thiazolyl blue tetrazolium. The affinity of the enzyme for 3-methyleneoxindole (Ks = 0.5 mm 3-methyleneoxindole) is relatively high. It is, therefore, reasonable to assume that 3-methyleneoxindole is the normal substrate. The enzyme is inhibited by indole-3-acetic acid, indole-3-aldehyde, and by l-naph-thaleneacetic acid. While these are not especially powerful inhibitors (K1 = 1.9-4.0 mm) the competitive relationship with 3-methyleneoxindole indicates that significant inhibition might occur at low intracellular concentrations of the substrate. PMID:6042360

  17. Redistribution of annexin in gravistimulated pea plumules

    NASA Technical Reports Server (NTRS)

    Clark, G. B.; Rafati, D. S.; Bolton, R. J.; Dauwalder, M.; Roux, S. J.

    2000-01-01

    We used immunocytochemistry to investigate the effects of gravistimulation on annexin localization in etiolated pea plumule shoots. In longitudinal sections, an asymmetric annexin immunostaining pattern was observed in a defined group of cells located just basipetal to apical meristems at the main shoot apex and at all of the axillary buds, an area classically referred to as the leaf gap. The pattern was observed using both protein-A-purified anti-annexin and affinity-purified anti-annexin antibodies for the immunostaining. A subset of the cells with the annexin staining also showed an unusually high level of periodic acid Schiff (PAS) staining in their cell walls. Prior to gravistimulation, the highest concentration of annexin was oriented toward the direction of gravity along the apical end of these immunostained cells. In contrast, both at 15 and 30 min after gravistimulation, the annexin immunostain became more evenly distributed all around the cell and more distinctly cell peripheral. The asymmetry along the lower wall of these cells was no longer evident. In accord with current models of annexin action, we interpret the results to indicate that annexin-mediated secretion in the leaf gap area is preferentially toward the apical meristem prior to gravistimulation, and that gravistimulation results in a redirection of this secretion. These data are to our knowledge the first to show a correlation between the vector of gravity and the distribution of annexins in the cells of flowering plants. c 2000 Editions scientifiques et medicales Elsevier SAS.

  18. Redistribution of annexin in gravistimulated pea plumules

    NASA Technical Reports Server (NTRS)

    Clark, G. B.; Rafati, D. S.; Bolton, R. J.; Dauwalder, M.; Roux, S. J.

    2000-01-01

    We used immunocytochemistry to investigate the effects of gravistimulation on annexin localization in etiolated pea plumule shoots. In longitudinal sections, an asymmetric annexin immunostaining pattern was observed in a defined group of cells located just basipetal to apical meristems at the main shoot apex and at all of the axillary buds, an area classically referred to as the leaf gap. The pattern was observed using both protein-A-purified anti-annexin and affinity-purified anti-annexin antibodies for the immunostaining. A subset of the cells with the annexin staining also showed an unusually high level of periodic acid Schiff (PAS) staining in their cell walls. Prior to gravistimulation, the highest concentration of annexin was oriented toward the direction of gravity along the apical end of these immunostained cells. In contrast, both at 15 and 30 min after gravistimulation, the annexin immunostain became more evenly distributed all around the cell and more distinctly cell peripheral. The asymmetry along the lower wall of these cells was no longer evident. In accord with current models of annexin action, we interpret the results to indicate that annexin-mediated secretion in the leaf gap area is preferentially toward the apical meristem prior to gravistimulation, and that gravistimulation results in a redirection of this secretion. These data are to our knowledge the first to show a correlation between the vector of gravity and the distribution of annexins in the cells of flowering plants. c 2000 Editions scientifiques et medicales Elsevier SAS.

  19. Protein methylation in pea chloroplasts. [Pisum sativum

    SciTech Connect

    Niemi, K.J.; Adler, J.; Selman, B.R. )

    1990-07-01

    The methylation of chloroplast proteins has been investigated by incubating intact pea (Pisum sativum) chloroplasts with ({sup 3}H-methyl)-S-adenosylmethionine. Incubation in the light increases the amount of methylation in both the thylakoid and stromal fractions. Numerous thylakoid proteins serve as substrates for the methyltransfer reactions. Three of these thylakoid proteins are methylated to a significantly greater extent in the light than in the dark. The primary stromal polypeptide methylated is the large subunit of ribulose bisphosphate carboxylase/oxygenase. One other stromal polypeptide is also methylated much more in the light than in the dark. Two distinct types of protein methylation occur. One methylinkage is stable to basic conditions whereas a second type is base labile. The base-stable linkage is indicative of N-methylation of amino acid residues while base-lability is suggestive of carboxymethylation of amino acid residues. Labeling in the light increases the percentage of methylation that is base labile in the thylakoid fraction while no difference is observed in the amount of base-labile methylations in light-labeled and dark-labeled stromal proteins. Also suggestive of carboxymethylation is the detection of volatile ({sup 3}H)methyl radioactivity which increases during the labeling period and is greater in chloroplasts labeled in the light as opposed to being labeled in the dark; this implies in vivo turnover of the ({sup 3}H)methyl group.

  20. Antifungal chromans inhibiting the mitochondrial respiratory chain of pea seeds and new xanthones from Calophyllum caledonicum.

    PubMed

    Hay, A-E; Guilet, D; Morel, C; Larcher, G; Macherel, D; Le Ray, A-M; Litaudon, M; Richomme, P

    2003-12-01

    Two new xanthones, caledonixanthone M 1 and caloxanthone L 2, and one new acid, caledonic acid 6 were isolated from the hexane-soluble extract of the stem bark of Calophyllum caledonicum. In the course of this phytochemical study, seven other known compounds - calothwaitesixanthone, calozeyloxanthone, allanxanthone, isoapetalic acid 3, calolongic acid 4, apetalic acid 5 and isocalolongic acid 7 - were isolated. Their antifungal activity against the growth of the human pathogenic fungus Aspergillus fumigatus was then investigated. The results indicated that the crude extract, calolongic acid 4 and isocalolongic acid 7 exhibited strong inhibitory effects with MIC (80) values of 8, 4, 2 microg/mL, respectively. Besides, calolongic acid 4, its lactone derivative 4a and isocalolongic acid 7 markedly reduced the respiration of pea seed mitochondria.

  1. Social Aggregation in Pea Aphids: Experiment and Random Walk Modeling

    PubMed Central

    Nilsen, Christa; Paige, John; Warner, Olivia; Mayhew, Benjamin; Sutley, Ryan; Lam, Matthew; Bernoff, Andrew J.; Topaz, Chad M.

    2013-01-01

    From bird flocks to fish schools and ungulate herds to insect swarms, social biological aggregations are found across the natural world. An ongoing challenge in the mathematical modeling of aggregations is to strengthen the connection between models and biological data by quantifying the rules that individuals follow. We model aggregation of the pea aphid, Acyrthosiphon pisum. Specifically, we conduct experiments to track the motion of aphids walking in a featureless circular arena in order to deduce individual-level rules. We observe that each aphid transitions stochastically between a moving and a stationary state. Moving aphids follow a correlated random walk. The probabilities of motion state transitions, as well as the random walk parameters, depend strongly on distance to an aphid's nearest neighbor. For large nearest neighbor distances, when an aphid is essentially isolated, its motion is ballistic with aphids moving faster, turning less, and being less likely to stop. In contrast, for short nearest neighbor distances, aphids move more slowly, turn more, and are more likely to become stationary; this behavior constitutes an aggregation mechanism. From the experimental data, we estimate the state transition probabilities and correlated random walk parameters as a function of nearest neighbor distance. With the individual-level model established, we assess whether it reproduces the macroscopic patterns of movement at the group level. To do so, we consider three distributions, namely distance to nearest neighbor, angle to nearest neighbor, and percentage of population moving at any given time. For each of these three distributions, we compare our experimental data to the output of numerical simulations of our nearest neighbor model, and of a control model in which aphids do not interact socially. Our stochastic, social nearest neighbor model reproduces salient features of the experimental data that are not captured by the control. PMID:24376691

  2. Social aggregation in pea aphids: experiment and random walk modeling.

    PubMed

    Nilsen, Christa; Paige, John; Warner, Olivia; Mayhew, Benjamin; Sutley, Ryan; Lam, Matthew; Bernoff, Andrew J; Topaz, Chad M

    2013-01-01

    From bird flocks to fish schools and ungulate herds to insect swarms, social biological aggregations are found across the natural world. An ongoing challenge in the mathematical modeling of aggregations is to strengthen the connection between models and biological data by quantifying the rules that individuals follow. We model aggregation of the pea aphid, Acyrthosiphon pisum. Specifically, we conduct experiments to track the motion of aphids walking in a featureless circular arena in order to deduce individual-level rules. We observe that each aphid transitions stochastically between a moving and a stationary state. Moving aphids follow a correlated random walk. The probabilities of motion state transitions, as well as the random walk parameters, depend strongly on distance to an aphid's nearest neighbor. For large nearest neighbor distances, when an aphid is essentially isolated, its motion is ballistic with aphids moving faster, turning less, and being less likely to stop. In contrast, for short nearest neighbor distances, aphids move more slowly, turn more, and are more likely to become stationary; this behavior constitutes an aggregation mechanism. From the experimental data, we estimate the state transition probabilities and correlated random walk parameters as a function of nearest neighbor distance. With the individual-level model established, we assess whether it reproduces the macroscopic patterns of movement at the group level. To do so, we consider three distributions, namely distance to nearest neighbor, angle to nearest neighbor, and percentage of population moving at any given time. For each of these three distributions, we compare our experimental data to the output of numerical simulations of our nearest neighbor model, and of a control model in which aphids do not interact socially. Our stochastic, social nearest neighbor model reproduces salient features of the experimental data that are not captured by the control.

  3. Total photoabsorption in nuclei

    SciTech Connect

    Bianchi, N.

    1992-06-01

    The Frascati-Genova collaboration proposes to measure the total photonuclear cross section on a wide range of nuclei between 500 MeV and 2 GeV, to obtain informations on the interaction of baryon resonances with nucleons and on the onset of the shadowing effect. The experiment could be performed in the Hall B as soon as the tagging facility will be ready and before the end of the installation of the CLAS spectrometer. The requirements for the photon beam, like maximum energy, intensity and beam definition, are not so strong so that the experiment would also be a good first test of the tagged photon facility.

  4. Electroproduction of Strange Nuclei

    SciTech Connect

    E.V. Hungerford

    2002-06-01

    The advent of high-energy, CW-beams of electrons now allows electro-production and precision studies of nuclei containing hyperons. Previously, the injection of strangeness into a nucleus was accomplished using secondary beams of mesons, where beam quality and target thickness limited the missing mass resolution. We review here the theoretical description of the (e, e'K+) reaction mechanism, and discuss the first experiment demonstrating that this reaction can be used to precisely study the spectra of light hypernuclei. Future experiments based on similar techniques, are expected to attain even better resolutions and rates.

  5. Lattice QCD for nuclei

    NASA Astrophysics Data System (ADS)

    Beane, Silas

    2016-09-01

    Over the last several decades, theoretical nuclear physics has been evolving from a very-successful phenomenology of the properties of nuclei, to a first-principles derivation of the properties of visible matter in the Universe from the known underlying theories of Quantum Chromodynamics (QCD) and Electrodynamics. Many nuclear properties have now been calculated using lattice QCD, a method for treating QCD numerically with large computers. In this talk, some of the most recent results in this frontier area of nuclear theory will be reviewed.

  6. Predictions for Superheavy Nuclei

    NASA Astrophysics Data System (ADS)

    Kumar, Krishna

    1990-01-01

    The Dynamic Deformation Model has been extended to the problem of fission in such a way that several thousand channels including particle-decay, α-decay, heavy-ion-emission, asymmetric fission, and symmetric fission can be taken into account. The model also includes a Kinetic Shell Correction which was ignored in previous predictions for Superheavy nuclei. This model is in better agreement with experimental life-times. A new location of the Superheavy peak is predicted at Z = 116 (eka-Polonium), A = 300, total half-life = 1079 years. New heavy-ion-fusion experiments and the means of identifying the Superheavy Elements are suggested.

  7. Genome sequence of the pea aphid Acyrthosiphon pisum.

    PubMed

    2010-02-23

    Aphids are important agricultural pests and also biological models for studies of insect-plant interactions, symbiosis, virus vectoring, and the developmental causes of extreme phenotypic plasticity. Here we present the 464 Mb draft genome assembly of the pea aphid Acyrthosiphon pisum. This first published whole genome sequence of a basal hemimetabolous insect provides an outgroup to the multiple published genomes of holometabolous insects. Pea aphids are host-plant specialists, they can reproduce both sexually and asexually, and they have coevolved with an obligate bacterial symbiont. Here we highlight findings from whole genome analysis that may be related to these unusual biological features. These findings include discovery of extensive gene duplication in more than 2000 gene families as well as loss of evolutionarily conserved genes. Gene family expansions relative to other published genomes include genes involved in chromatin modification, miRNA synthesis, and sugar transport. Gene losses include genes central to the IMD immune pathway, selenoprotein utilization, purine salvage, and the entire urea cycle. The pea aphid genome reveals that only a limited number of genes have been acquired from bacteria; thus the reduced gene count of Buchnera does not reflect gene transfer to the host genome. The inventory of metabolic genes in the pea aphid genome suggests that there is extensive metabolite exchange between the aphid and Buchnera, including sharing of amino acid biosynthesis between the aphid and Buchnera. The pea aphid genome provides a foundation for post-genomic studies of fundamental biological questions and applied agricultural problems.

  8. Genome Sequence of the Pea Aphid Acyrthosiphon pisum

    PubMed Central

    2010-01-01

    Aphids are important agricultural pests and also biological models for studies of insect-plant interactions, symbiosis, virus vectoring, and the developmental causes of extreme phenotypic plasticity. Here we present the 464 Mb draft genome assembly of the pea aphid Acyrthosiphon pisum. This first published whole genome sequence of a basal hemimetabolous insect provides an outgroup to the multiple published genomes of holometabolous insects. Pea aphids are host-plant specialists, they can reproduce both sexually and asexually, and they have coevolved with an obligate bacterial symbiont. Here we highlight findings from whole genome analysis that may be related to these unusual biological features. These findings include discovery of extensive gene duplication in more than 2000 gene families as well as loss of evolutionarily conserved genes. Gene family expansions relative to other published genomes include genes involved in chromatin modification, miRNA synthesis, and sugar transport. Gene losses include genes central to the IMD immune pathway, selenoprotein utilization, purine salvage, and the entire urea cycle. The pea aphid genome reveals that only a limited number of genes have been acquired from bacteria; thus the reduced gene count of Buchnera does not reflect gene transfer to the host genome. The inventory of metabolic genes in the pea aphid genome suggests that there is extensive metabolite exchange between the aphid and Buchnera, including sharing of amino acid biosynthesis between the aphid and Buchnera. The pea aphid genome provides a foundation for post-genomic studies of fundamental biological questions and applied agricultural problems. PMID:20186266

  9. Characterization of pea (Pisum sativum) seed protein fractions.

    PubMed

    Rubio, Luis A; Pérez, Alicia; Ruiz, Raquel; Guzmán, M Ángeles; Aranda-Olmedo, Isabel; Clemente, Alfonso

    2014-01-30

    Legume seed proteins have to be chemically characterized in order to properly link their nutritional effects with their chemical structure. Vicilin and albumin fractions devoid of cross-contamination, as assessed by mass peptide fingerprinting analysis, were obtained from defatted pea (Pisum sativum cv. Bilbo) meal. The extracted protein fractions contained 56.7-67.7 g non-starch polysaccharides kg⁻¹. The vicilin fraction was higher than legumins in arginine, isoleucine, leucine, phenylalanine and lysine. The most abundant amino acids in the albumin fraction were aspartic acid, glutamic acid, lysine and arginine, and the amounts of methionine were more than double than those in legumins and vicilins. The pea albumin fraction showed a clear enrichment of protease inhibitory activity when compared with the seed meal. In vitro digestibility values for pea proteins were 0.63 ±  0.04, 0.88 ±  0.04 and 0.41 ±  0.23 for legumins, vicilins and albumins respectively. Vicilin and albumin fractions devoid of cross-contamination with other proteins were obtained from pea seed meal. The vicilin fraction also contained low amounts of soluble non-starch polysaccharides and was enriched in isoleucine, leucine, phenylalanine and lysine. In vitro digestibility values for pea proteins were similar or even numerically higher than those for control proteins. © 2013 Society of Chemical Industry.

  10. Activity of nicotinamide–adenine dinucleotide pyrophosphorylase in liver nuclei. Effects of partial hepatectomy, hepatotoxins and dietary changes

    PubMed Central

    Stirpe, F.; Corte, E. Della

    1968-01-01

    1. The activity of NAD pyrophosphorylase is lower in nuclei isolated from regenerating rat liver than in normal nuclei, and this is due to leakage of the enzyme from the nuclei during the isolation. 2. The NAD pyrophosphorylase activity is lower in liver nuclei from newborn rats, and from rats on a protein-free diet, but no leakage occurs in these cases. 3. Poisoning with α-amanitin brings about a transient enhancement of NAD pyrophosphorylase activity in mouse liver nuclei. 4. No changes of enzyme activity were observed after 72hr. starvation, administration of actinomycin D or infection with MHV3 virus. PMID:4297940

  11. D mesic nuclei

    NASA Astrophysics Data System (ADS)

    García-Recio, C.; Nieves, J.; Tolos, L.

    2010-06-01

    The energies and widths of several D0 meson bound states for different nuclei are obtained using a D-meson selfenergy in the nuclear medium, which is evaluated in a selfconsistent manner using techniques of unitarized coupled-channel theory. The kernel of the meson-baryon interaction is based on a model that treats heavy pseudoscalar and heavy vector mesons on equal footing, as required by heavy quark symmetry. We find D0 bound states in all studied nuclei, from 12C up to 208Pb. The inclusion of vector mesons is the keystone for obtaining an attractive D-nucleus interaction that leads to the existence of D0-nucleus bound states, as compared to previous studies based on SU(4) flavor symmetry. In some cases, the half widths are smaller than the separation of the levels, what makes possible their experimental observation by means of a nuclear reaction. This can be of particular interest for the future P¯ANDA@FAIR physics program. We also find a D+ bound state in 12C, but it is too broad and will have a significant overlap with the energies of the continuum.

  12. Potential alternative hosts for the pea powdery mildew pathogen Erysiphe trifolii

    USDA-ARS?s Scientific Manuscript database

    Powdery mildew of pea (Pisum sativum) is an important disease in the field and in the greenhouse. The most widely documented powdery mildew pathogen on pea is Erysiphe pisi, but E. baeumleri and E. trifolii have also been reported. We recently showed that E. trifolii is frequently found on pea in th...

  13. 76 FR 37136 - Post-Entry Amendment (PEA) Processing Test: Modification, Clarification, and Extension

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-24

    ... SECURITY U.S. Customs and Border Protection Post-Entry Amendment (PEA) Processing Test: Modification... (CBP's) Post-Entry Amendment (PEA) Processing test, which allows the amendment of entry summaries prior to liquidation. The test is being modified to reflect that PEA procedures will no longer be accepted...

  14. The International Pea Genome Sequencing Project: Sequencing and Assembly Progresses Updates

    USDA-ARS?s Scientific Manuscript database

    The International Consortium for the Pea Genome Sequencing (ICPG) includes scientists from six countries around the world. Its aim is to provide a high quality reference of the pea genome to the scientific community as well as to the pea breeder community. The consortium proposed a strategy that int...

  15. Differences in weed seedling emergence is not involved in pea synergism to corn

    USDA-ARS?s Scientific Manuscript database

    We have observed the dry pea is synergistic to corn and improves its tolerance to weeds. We are examining various aspects of this interaction between dry pea and corn to understand this natural benefit. This study compared the impact of soybean and dry pea on corn growth and tolerance to foxtail m...

  16. Heavy and Superheavy Atomic Nuclei

    NASA Astrophysics Data System (ADS)

    Sobiczewski, Adam

    2008-10-01

    The appearance and development of the concept of super-heavy atomic nuclei are described. The concept appeared during the studies of the limits of the nuclear chart and of the periodic table of the chemical elements. The article concentrates on theoretical studies of the properties of heaviest nuclei. Results of these studies are illustrated and discussed. Prospects for a nearest future of the research of heaviest nuclei are outlined.

  17. Light-modulated abundance of an mRNA encoding a calmodulin-regulated, chromatin-associated NTPase in pea

    NASA Technical Reports Server (NTRS)

    Hsieh, H. L.; Tong, C. G.; Thomas, C.; Roux, S. J.

    1996-01-01

    A CDNA encoding a 47 kDa nucleoside triphosphatase (NTPase) that is associated with the chromatin of pea nuclei has been cloned and sequenced. The translated sequence of the cDNA includes several domains predicted by known biochemical properties of the enzyme, including five motifs characteristic of the ATP-binding domain of many proteins, several potential casein kinase II phosphorylation sites, a helix-turn-helix region characteristic of DNA-binding proteins, and a potential calmodulin-binding domain. The deduced primary structure also includes an N-terminal sequence that is a predicted signal peptide and an internal sequence that could serve as a bipartite-type nuclear localization signal. Both in situ immunocytochemistry of pea plumules and immunoblots of purified cell fractions indicate that most of the immunodetectable NTPase is within the nucleus, a compartment proteins typically reach through nuclear pores rather than through the endoplasmic reticulum pathway. The translated sequence has some similarity to that of human lamin C, but not high enough to account for the earlier observation that IgG against human lamin C binds to the NTPase in immunoblots. Northern blot analysis shows that the NTPase MRNA is strongly expressed in etiolated plumules, but only poorly or not at all in the leaf and stem tissues of light-grown plants. Accumulation of NTPase mRNA in etiolated seedlings is stimulated by brief treatments with both red and far-red light, as is characteristic of very low-fluence phytochrome responses. Southern blotting with pea genomic DNA indicates the NTPase is likely to be encoded by a single gene.

  18. Light-modulated abundance of an mRNA encoding a calmodulin-regulated, chromatin-associated NTPase in pea

    NASA Technical Reports Server (NTRS)

    Hsieh, H. L.; Tong, C. G.; Thomas, C.; Roux, S. J.

    1996-01-01

    A CDNA encoding a 47 kDa nucleoside triphosphatase (NTPase) that is associated with the chromatin of pea nuclei has been cloned and sequenced. The translated sequence of the cDNA includes several domains predicted by known biochemical properties of the enzyme, including five motifs characteristic of the ATP-binding domain of many proteins, several potential casein kinase II phosphorylation sites, a helix-turn-helix region characteristic of DNA-binding proteins, and a potential calmodulin-binding domain. The deduced primary structure also includes an N-terminal sequence that is a predicted signal peptide and an internal sequence that could serve as a bipartite-type nuclear localization signal. Both in situ immunocytochemistry of pea plumules and immunoblots of purified cell fractions indicate that most of the immunodetectable NTPase is within the nucleus, a compartment proteins typically reach through nuclear pores rather than through the endoplasmic reticulum pathway. The translated sequence has some similarity to that of human lamin C, but not high enough to account for the earlier observation that IgG against human lamin C binds to the NTPase in immunoblots. Northern blot analysis shows that the NTPase MRNA is strongly expressed in etiolated plumules, but only poorly or not at all in the leaf and stem tissues of light-grown plants. Accumulation of NTPase mRNA in etiolated seedlings is stimulated by brief treatments with both red and far-red light, as is characteristic of very low-fluence phytochrome responses. Southern blotting with pea genomic DNA indicates the NTPase is likely to be encoded by a single gene.

  19. Scissors mode of Gd nuclei studied from resonance neutron capture

    SciTech Connect

    Kroll, J.; Baramsai, B.; Becker, J. A.; and others

    2012-10-20

    Spectra of {gamma} rays following the neutron capture at isolated resonances of stable Gd nuclei were measured. The objectives were to get new information on photon strength of {sup 153,155-159}Gd with emphasis on the role of the M1 scissors-mode vibration. An analysis of the data obtained clearly indicates that the scissors mode is coupled not only to the ground state, but also to all excited levels of the nuclei studied. The specificity of our approach ensures unbiasedness in estimating the sumed scissors-mode strength {Sigma}B(M1){up_arrow}, even for odd product nuclei, for which conventional nuclear resonance fluorescence measurements yield only limited information. Our analysis indicates that for these nuclei the sum {Sigma}B(M1){up_arrow} increases with A and for {sup 157,159}Gd it is significantly higher compared to {sup 156,158}Gd.

  20. Host Adaptation of Soybean Dwarf Virus Following Serial Passages on Pea (Pisum sativum) and Soybean (Glycine max).

    PubMed

    Tian, Bin; Gildow, Frederick E; Stone, Andrew L; Sherman, Diana J; Damsteegt, Vernon D; Schneider, William L

    2017-06-21

    Soybean Dwarf Virus (SbDV) is an important plant pathogen, causing economic losses in soybean. In North America, indigenous strains of SbDV mainly infect clover, with occasional outbreaks in soybean. To evaluate the risk of a US clover strain of SbDV adapting to other plant hosts, the clover isolate SbDV-MD6 was serially transmitted to pea and soybean by aphid vectors. Sequence analysis of SbDV-MD6 from pea and soybean passages identified 11 non-synonymous mutations in soybean, and six mutations in pea. Increasing virus titers with each sequential transmission indicated that SbDV-MD6 was able to adapt to the plant host. However, aphid transmission efficiency on soybean decreased until the virus was no longer transmissible. Our results clearly demonstrated that the clover strain of SbDV-MD6 is able to adapt to soybean crops. However, mutations that improve replication and/or movement may have trade-off effects resulting in decreased vector transmission.

  1. Amino Acid Transporters Are Localized to Transfer Cells of Developing Pea Seeds1

    PubMed Central

    Tegeder, Mechthild; Offler, Christina E.; Frommer, Wolf B.; Patrick, John W.

    2000-01-01

    To determine the nature and cellular localization of amino acid transport in pea seeds, two cDNA clones belonging to the AAP family of H+/amino acid co-transporters (PsAAP1 and PsAAP2) were isolated from a cotyledon cDNA library of pea (Pisum sativum L.). Functional expression in the yeast amino acid uptake mutants 22Δ6AAL and 22Δ8AA showed that PsAAP1 mediates transport of neutral, acidic, and basic amino acids. RNA-blot analyses showed that PsAAP1 is expressed in seeds and vegetative organs, including amino acid sinks and sources, whereas PsAAP2 could not be detected. For developing seeds, transcripts of PsAAP1 were detected in coats and cotyledons, with seed coats giving a weak signal. In cotyledons, expression was highest in epidermal-transfer-cell-enriched tissue. RNA in situ hybridization analysis showed that PsAAP1 was predominantly present in epidermal transfer cells forming the outer surface of cotyledons, which abuts the seed coats. Overall, our observations suggest that this transporter, which is localized in transfer cells of cotyledons, might play a role in the uptake of the full spectrum of amino acids released from seed coats. PMID:10677425

  2. Separation and Partial Characterization of Two Ribonucleic Acid Polymerases from Pea Seedlings 1

    PubMed Central

    Glicklich, Daniel; Jendrisak, Jerome J.; Becker, Wayne M.

    1974-01-01

    Two DNA-dependent RNA polymerases (ribonucleoside triphosphate:RNA nucleotidyl transferase, EC 2.7.7.6) have been isolated from pea (Pisum sativum) seedlings. The enzymes were solubilized by sonication in high salt buffer and were separated by chromatography on diethylaminoethyl cellulose using a linear salt gradient. Polymerase I eluted at 0.10 m (NH4)2SO4, accounted for about 10% of the recovered activity and was completely insensitive to α-amanitin. Polymerase II eluted at 0.14 m (NH4)2SO4, accounted for the remaining 90% of recovered activity and was strongly inhibited by α-amanitin. Both enzymes preferred denatured to native DNA as template, both showed an absolute requirement of divalent cation, and both were sensitive to the ionic strength of the assay medium. The developing pea seedling seems a promising system for studies of possible changes in relative activities and roles of multiple RNA polymerases during eukaryotic development. PMID:16658887

  3. Adaptation of photosynthetic electron-transport rate to growth temperature in pea.

    PubMed

    Mitchell, R A; Barber, J

    1986-11-01

    Pea (Pisum sativum L. cv. Feltham First) plants were germinated and grown under two temperature regimes, one chilling (6-8° C) and one non-chilling (16-18° C), which are referred to as "cold-grown" and "warm-grown", respectively. It was found that: (1) At saturating light intensity and with excess CO2, cold-grown leaves exhibited faster rates of oxygen evolution than warm-grown leaves when measured below 15° C. However when measurements were carried out above this temperature, the reverse relationship was observed. (2) Full-chain electron-transport measurements on thylakoids showed that those isolated from cold-grown plants had greater light-saturated uncoupled rates than their warm-grown equivalents at all temperatures between 3 and 19° C. (3) This difference was apparently not due to a greater activity of photosystem I or II in the thylakoids from cold-grown plants, but rather to a more rapid turnover of a dark step within the electron-transport chain. These results are interpreted in terms of a previously reported apparent homeoviscous adaptation of the pea thylakoid membrane to growth temperature (J. Barber, R.C. Ford, R.A.C. Mitchell, P.A. Millner, 1984, Planta 161, 375-380).

  4. Expression of ribosomal genes in pea cotyledons at the initial stages of germination

    SciTech Connect

    Gumilevskaya, N.A.; Chumikhina, L.V.; Akhmatova, A.T.; Kretovich, V.L.

    1986-01-20

    The time of appearance of newly synthesized rRNAs and ribosomal proteins (r-proteins) in the ribosomes of pea cotyledons (Pisum sativum L.) during germination was investigated. The ribosomal fraction was isolated and analyzed according to the method of germination of the embryo in the presence of labeled precursors or after pulse labeling of the embryos at different stages of germination. For the identification of newly synthesized rRNAs in the ribosomes we estimated the relative stability of labeled RNAs to the action of RNase, the sedimentation rate, the ability to be methylated in vivo in the presence of (/sup 14/C)CH/sub 3/-methionine, and the localization in the subunits of dissociated ribosomes. The presence of newly synthesized r-proteins in the ribosomes was judged on the basis of the electrophoretic similarity in SDS-disc electrophoresis of labeled polypeptides of purified ribosome preparations and of genuine r-proteins, as well as according to the localization of labeled proteins in the subunits of the dissociated ribosomes. It was shown that the expression of the ribosomal genes in highly specialized cells of pea cotyledons that have completed their growth occurs at very early stages of germination.

  5. Differential gene expression according to race and host plant in the pea aphid.

    PubMed

    Eyres, Isobel; Jaquiéry, Julie; Sugio, Akiko; Duvaux, Ludovic; Gharbi, Karim; Zhou, Jing-Jiang; Legeai, Fabrice; Nelson, Michaela; Simon, Jean-Christophe; Smadja, Carole M; Butlin, Roger; Ferrari, Julia

    2016-09-01

    Host-race formation in phytophagous insects is thought to provide the opportunity for local adaptation and subsequent ecological speciation. Studying gene expression differences amongst host races may help to identify phenotypes under (or resulting from) divergent selection and their genetic, molecular and physiological bases. The pea aphid (Acyrthosiphon pisum) comprises host races specializing on numerous plants in the Fabaceae and provides a unique system for examining the early stages of diversification along a gradient of genetic and associated adaptive divergence. In this study, we examine transcriptome-wide gene expression both in response to environment and across pea aphid races selected to cover the range of genetic divergence reported in this species complex. We identify changes in expression in response to host plant, indicating the importance of gene expression in aphid-plant interactions. Races can be distinguished on the basis of gene expression, and higher numbers of differentially expressed genes are apparent between more divergent races; these expression differences between host races may result from genetic drift and reproductive isolation and possibly divergent selection. Expression differences related to plant adaptation include a subset of chemosensory and salivary genes. Genes showing expression changes in response to host plant do not make up a large portion of between-race expression differences, providing confirmation of previous studies' findings that genes involved in expression differences between diverging populations or species are not necessarily those showing initial plasticity in the face of environmental change.

  6. Influence of ammonium chloride on the nitrogenase activity of nodulated pea plants (Pisum sativum).

    PubMed Central

    Houwaard, F

    1978-01-01

    A study was made on the short-term effect of ammonium ions on the nitrogenase activity of pea root nodules. Nodulated pea plants (Pisum sativum), having reached maximum acetylene-reducing activity, were supplied with NH4Cl (20 mM). Nitrogenase activity of intact plants, detached nodules, and isolated bacteroids was measured at differed time intervals. A significant drop (20 to 40%) in the acetylene-reducing activity of treated intact plants and their detached nodules was observed after 1 day. No drop in the nitrogenase activity of bacteroids (assayed aerobically, or anaerobically after treatment with ethylenediaminetetraacetic acid-toluene) occurred for 2 to 4 days after the addition of NH4+ to the plants, depending on cultural conditions. From these results it is concluded that the adverse effect of NH4+ on acetylene reduction by intact plants and detached nodules during the first 2 days is not due to a decrease in the amount of nitrogenase in the bacteroids. It is suggested that the effect has to be attributed to a reduced supply to the bacteroids of energy-delivery photosynthates. PMID:677873

  7. Properties of Plasma Membrane from Pea Root Seedlings under Altered Gravity

    NASA Astrophysics Data System (ADS)

    Klymchuk, D.; Baranenko, V.; Vorobyova, T. V.; Kurylenko, I.; Chyzhykova, O.; Dubovoy, V.

    In this study, the properties of pea (Pisum sativum L.) plasma membrane were examined to determine how the membrane structure and functions are regulated in response to clinorotation (2 rev/min) conditions. Membrane preparations enriched by plasma membrane vesicles were obtained by aqueous two-phase partitioning from 6-day seedling roots. The specific characteristics of H^+-ATPase, lípid composition and peroxidation intensity as well as fluidity of lipid bilayer were analysed. ATP hydrolytic activity was inhibited by ortovanadate and was insensitive to aside and nitrate in sealed plasma membrane vesicles isolated from both clinorotated and control seedlings. Plasma membrane vesicles from clinorotated seedlings in comparison to controls were characterised by increase in the total lipid/protein ratio, ATP hydrolytic activity and intensifying of lipid peroxidation. Sitosterol and campesterol were the predominant free sterol species. Clinorotated seedlings contained a slightly higher level of unsaturated fatty acid than controls. Plasma membrane vesicles were labelled with pyrene and fluorescence originating from monomeric (I_M) molecules and excimeric (I_E) aggregates were measured. The calculated I_E/I_M values were higher in clinorotated seedlings compared with controls reflecting the reduction in membrane microviscosity. The involvement of the changes in plasma membrane lipid content and composition, fluidity and H^+-ATPase activity in response of pea seedlings to altered gravity is discussed.

  8. PsPMEP, a pollen-specific pectin methylesterase of pea (Pisum sativum L.).

    PubMed

    Gómez, María Dolores; Renau-Morata, Begoña; Roque, Edelín; Polaina, Julio; Beltrán, José Pío; Cañas, Luis A

    2013-09-01

    Pectin methylesterases (PMEs) are a family of enzymes involved in plant reproductive processes such as pollen development and pollen tube growth. We have isolated and characterized PsPMEP, a pea (Pisum sativum L.) pollen-specific gene that encodes a protein with homology to PMEs. Sequence analysis showed that PsPMEP belongs to group 2 PMEs, which are characterized by the presence of a processable amino-terminal PME inhibitor domain followed by the catalytic PME domain. Moreover, PsPMEP contains several motifs highly conserved among PMEs with the essential amino acid residues involved in enzyme substrate binding and catalysis. Northern blot and in situ hybridization analyses showed that PsPMEP is expressed in pollen grains from 4 days before anthesis till anther dehiscence and in pollinated carpels. In the PsPMEP promoter region, we have identified several conserved cis-regulatory elements that have been associated with gene pollen-specific expression. Expression analysis of PsPMEP promoter fused to the uidA reporter gene in Arabidopsis thaliana plants showed a similar expression pattern when compared with pea, indicating that this promoter is also functional in a non-leguminous plant. GUS expression was detected in mature pollen grains, during pollen germination, during pollen tube elongation along the transmitting tract, and when the pollen tube reaches the embryo sac in the ovule.

  9. Pea DNA helicase 45 overexpression in tobacco confers high salinity tolerance without affecting yield.

    PubMed

    Sanan-Mishra, Neeti; Pham, Xuan Hoi; Sopory, Sudhir K; Tuteja, Narendra

    2005-01-11

    Salt tolerance is an important trait that is required to overcome salinity-induced reduction in plant productivity. We have reported previously the isolation of a pea DNA helicase 45 (PDH45) that exhibits striking homology with the eukaryotic translation initiation factor eIF-4A. Here, we report that PDH45 mRNA is induced in pea seedlings in response to high salt, and its overexpression driven by a constitutive cauliflower mosaic virus-(35)S promoter in tobacco plants confers salinity tolerance, thus suggesting a previously undescribed pathway for manipulating stress tolerance in crop plants. The T(0) transgenic plants showed high levels of PDH45 protein in normal and stress conditions, as compared with WT plants. The T(0) transgenics also showed tolerance to high salinity as tested by a leaf disk senescence assay. The T(1) transgenics were able to grow to maturity and set normal viable seeds under continuous salinity stress without any reduction in plant yield in terms of seed weight. Measurement of Na(+) ions in different parts of the plant showed higher accumulation in the old leaves and negligible accumulation in seeds of T(1) transgenic lines as compared with the WT plants. The possible mechanism of salinity tolerance is discussed. Overexpression of PDH45 provides a possible example of the exploitation of DNA/RNA unwinding pathways for engineering salinity tolerance without affecting yield in crop plants.

  10. Separation of nuclei representing different phases of the growth cycle from unsynchronized mammalian cell cultures.

    PubMed

    McBride, O W; Peterson, E A

    1970-10-01

    Nuclei have been isolated from unsynchronized cultures of Chinese hamster fibroblasts after varying intervals of growth following the incorporation of thymidine (-3)H for 20 min. These nuclei were fractionated by unit gravity sedimentation in a stabilizing density gradient of sucrose, and fractions were analyzed for the concentration of nuclei, DNA, and radioactivity. A more rapidly sedimenting population of nuclei in the G(2) phase of the cell cycle was separated from a group of nuclei in the G(1) phase, and nuclei in progressive stages of DNA synthesis (S phase) were distributed between these two regions. The fractionation of intact cells by sedimentation according to their position in the cell cycle was found to be less satisfactory than the corresponding separation of nuclei. This probably results from the continuous accumulation of mass within individual cells throughout the entire cell cycle, whereas most of the mass of a nucleus is replicated during a relatively narrow interval of the total cell cycle.

  11. Exotic nuclei in astrophysics

    NASA Astrophysics Data System (ADS)

    Penionzhkevich, Yu. E.

    2012-07-01

    Recently the academic community has marked several anniversaries connected with discoveries that played a significant role in the development of astrophysical investigations. The year 2009 was proclaimed by the United Nations the International Year of Astronomy. This was associated with the 400th anniversary of Galileo Galilei's discovery of the optical telescope, which marked the beginning of regular research in the field of astronomy. An important contribution to not only the development of physics of the microcosm, but also to the understanding of processes occurring in the Universe, was the discovery of the atomic nucleus made by E. Rutherford 100 years ago. Since then the investigations in the fields of physics of particles and atomic nuclei have helped to understand many processes in the microcosm. Exactly 80 years ago, K. Yanski used a radio-telescope in order to receive the radiation from cosmic objects for the first time, and at the present time this research area of physics is the most efficient method for studying the properties of the Universe. Finally, the April 12, 1961 (50 years ago) launching of the first sputnik into space with a human being onboard, the Russian cosmonaut Yuri Gagarin, marked the beginning of exploration of the Universe with the direct participation of man. All these achievements considerably extended our ideas about the Universe. This work is an attempt to present some problems on the evolution of the Universe: the nucleosynthesis and cosmochronology from the standpoint of physics of particles and nuclei, in particular with the use of the latest results, obtained by means of radioactive nuclear beams. The comparison is made between the processes taking place in the Universe and the mechanisms of formation and decay of nuclei, as well as of their interaction at different energies. Examples are given to show the capabilities of nuclear-physics methods for studying cosmic objects and properties of the Universe. The results of

  12. Comparative Transcriptomic Analyses of Vegetable and Grain Pea (Pisum sativum L.) Seed Development

    PubMed Central

    Liu, Na; Zhang, Guwen; Xu, Shengchun; Mao, Weihua; Hu, Qizan; Gong, Yaming

    2015-01-01

    Understanding the molecular mechanisms regulating pea seed developmental process is extremely important for pea breeding. In this study, we used high-throughput RNA-Seq and bioinformatics analyses to examine the changes in gene expression during seed development in vegetable pea and grain pea, and compare the gene expression profiles of these two pea types. RNA-Seq generated 18.7 G of raw data, which were then de novo assembled into 77,273 unigenes with a mean length of 930 bp. Our results illustrate that transcriptional control during pea seed development is a highly coordinated process. There were 459 and 801 genes differentially expressed at early and late seed maturation stages between vegetable pea and grain pea, respectively. Soluble sugar and starch metabolism related genes were significantly activated during the development of pea seeds coinciding with the onset of accumulation of sugar and starch in the seeds. A comparative analysis of genes involved in sugar and starch biosynthesis in vegetable pea (high seed soluble sugar and low starch) and grain pea (high seed starch and low soluble sugar) revealed that differential expression of related genes at late development stages results in a negative correlation between soluble sugar and starch biosynthetic flux in vegetable and grain pea seeds. RNA-Seq data was validated by using real-time quantitative RT-PCR analysis for 30 randomly selected genes. To our knowledge, this work represents the first report of seed development transcriptomics in pea. The obtained results provide a foundation to support future efforts to unravel the underlying mechanisms that control the developmental biology of pea seeds, and serve as a valuable resource for improving pea breeding. PMID:26635856

  13. Nucleomorphs: enslaved algal nuclei.

    PubMed

    Cavalier-Smith, T

    2002-12-01

    Nucleomorphs of cryptomonad and chlorarachnean algae are the relict, miniaturised nuclei of formerly independent red and green algae enslaved by separate eukaryote hosts over 500 million years ago. The complete 551 kb genome sequence of a cryptomonad nucleomorph confirms that cryptomonads are eukaryote-eukaryote chimeras and greatly illuminates the symbiogenetic event that created the kingdom Chromista and their alveolate protozoan sisters. Nucleomorph membranes may, like plasma membranes, be more enduring after secondary symbiogenesis than are their genomes. Partial sequences of chlorarachnean nucleomorphs indicate that genomic streamlining is limited by the mutational difficulty of removing useless introns. Nucleomorph miniaturisation emphasises that selection can dramatically reduce eukaryote genome size and eliminate most non-functional nuclear non-coding DNA. Given the differential scaling of nuclear and nucleomorph genomes with cell size, it follows that most non-coding nuclear DNA must have a bulk-sequence-independent function related to cell volume.

  14. Pulsars:. Gigantic Nuclei

    NASA Astrophysics Data System (ADS)

    Xu, Renxin

    What is the real nature of pulsars? This is essentially a question of the fundamental strong interaction between quarks at low-energy scale and hence of the non-perturbative quantum chromo-dynamics, the solution of which would certainly be meaningful for us to understand one of the seven millennium prize problems (i.e., "Yang-Mills Theory") named by the Clay Mathematical Institute. After a historical note, it is argued here that a pulsar is very similar to an extremely big nucleus, but is a little bit different from the gigantic nucleus speculated 80 years ago by L. Landau. The paper demonstrates the similarity between pulsars and gigantic nuclei from both points of view: the different manifestations of compact stars and the general behavior of the strong interaction.

  15. Protein loss during nuclear isolation

    PubMed Central

    1983-01-01

    Cryomicrodissection makes possible the measurement of the entire in vivo protein content of the amphibian oocyte nucleus and provides a heretofore missing baseline for estimating protein loss during nuclear isolation by other methods. When oocyte nuclei are isolated into an aqueous medium, they lose 95% of their protein with a half-time of 250 s. This result implies an even more rapid loss of protein from aqueously isolated nuclei of ordinary-size cells. PMID:6619193

  16. Compton scattering by nuclei

    NASA Astrophysics Data System (ADS)

    Hütt, M.-Th.; L'vov, A. I.; Milstein, A. I.; Schumacher, M.

    2000-01-01

    The concept of Compton scattering by even-even nuclei from giant-resonance to nucleon-resonance energies and the status of experimental and theoretical researches in this field are outlined. The description of Compton scattering by nuclei starts from different complementary approaches, namely from second-order S-matrix and from dispersion theories. Making use of these, it is possible to incorporate into the predicted nuclear scattering amplitudes all the information available from other channels, viz. photon-nucleon and photon-meson channels, and to efficiently make use of models of the nucleon, the nucleus and the nucleon-nucleon interaction. The total photoabsorption cross section constrains the nuclear scattering amplitude in the forward direction. The specific information obtained from Compton scattering therefore stems from the angular dependence of the nuclear scattering amplitude, providing detailed insight into the dynamics of the nuclear and nucleon degrees of freedom and into the interplay between them. Nuclear Compton scattering in the giant-resonance energy-region provides information on the dynamical properties of the in-medium mass of the nucleon. Most prominently, the electromagnetic polarizabilities of the nucleon in the nuclear medium can be extracted from nuclear Compton scattering data obtained in the quasi-deuteron energy-region. In our description of this latter process special emphasis is laid upon the exploration of many-body and two-body effects entering into the nuclear dynamics. Recent results are presented for two-body effects due to the mesonic seagull amplitude and due to the excitation of nucleon internal degrees of freedom accompanied by meson exchanges. Due to these studies the in-medium electromagnetic polarizabilities are by now well understood, whereas the understanding of nuclear Compton scattering in the Δ-resonance range is only at the beginning. Furthermore, phenomenological methods how to include retardation effects in the

  17. Enhancing Neoplasm Expression in Field Pea (Pisum sativum) via Intercropping and Its Significance to Pea Weevil (Bruchus pisorum) Management

    PubMed Central

    Teshome, Abel; Bryngelsson, Tomas; Mendesil, Esayas; Marttila, Salla; Geleta, Mulatu

    2016-01-01

    Neoplasm formation, a non-meristematic tissue growth on young field pea (Pisum sativum L.) pods is triggered in the absence of UV light and/or in response to oviposition by pea weevil (Bruchus pisorum L.). This trait is expressed in some genotypes [neoplastic (Np) genotypes] of P. sativum and has the capacity to obstruct pea weevil larval entry into developing seeds. In the present study, 26% of the tested accessions depicted the trait when grown under greenhouse conditions. However, UV light inhibits full expression of this trait and subsequently it is inconspicuous at the field level. In order to investigate UV light impact on the expression of neoplasm, particular Np genotypes were subjected to UV lamp light exposure in the greenhouse and sunlight at the field level. Under these different growing conditions, the highest mean percentage of Np pods was in the control chamber in the greenhouse (36%) whereas in single and double UV lamp chambers, the percentage dropped to 10 and 15%, respectively. Furthermore, when the same Np genotypes were grown in the field, the percentage of Np pods dropped significantly (7%). In order to enhance Np expression at the field level, intercropping of Np genotypes with sorghum was investigated. As result, the percentage of Np pods was threefold in intercropped Np genotypes as compared to those without intercropping. Therefore, intercropping Np genotypes with other crops such as sorghum and maize can facilitate neoplasm formation, which in turn can minimize the success rate of pea weevil larvae entry into developing seeds. Greenhouse artificial infestation experiments showed that pea weevil damage in Np genotypes is lower in comparison to wild type genotypes. Therefore, promoting Np formation under field conditions via intercropping can serve as part of an integrated pea weevil management strategy especially for small scale farming systems. PMID:27242855

  18. Enhancing Neoplasm Expression in Field Pea (Pisum sativum) via Intercropping and Its Significance to Pea Weevil (Bruchus pisorum) Management.

    PubMed

    Teshome, Abel; Bryngelsson, Tomas; Mendesil, Esayas; Marttila, Salla; Geleta, Mulatu

    2016-01-01

    Neoplasm formation, a non-meristematic tissue growth on young field pea (Pisum sativum L.) pods is triggered in the absence of UV light and/or in response to oviposition by pea weevil (Bruchus pisorum L.). This trait is expressed in some genotypes [neoplastic (Np) genotypes] of P. sativum and has the capacity to obstruct pea weevil larval entry into developing seeds. In the present study, 26% of the tested accessions depicted the trait when grown under greenhouse conditions. However, UV light inhibits full expression of this trait and subsequently it is inconspicuous at the field level. In order to investigate UV light impact on the expression of neoplasm, particular Np genotypes were subjected to UV lamp light exposure in the greenhouse and sunlight at the field level. Under these different growing conditions, the highest mean percentage of Np pods was in the control chamber in the greenhouse (36%) whereas in single and double UV lamp chambers, the percentage dropped to 10 and 15%, respectively. Furthermore, when the same Np genotypes were grown in the field, the percentage of Np pods dropped significantly (7%). In order to enhance Np expression at the field level, intercropping of Np genotypes with sorghum was investigated. As result, the percentage of Np pods was threefold in intercropped Np genotypes as compared to those without intercropping. Therefore, intercropping Np genotypes with other crops such as sorghum and maize can facilitate neoplasm formation, which in turn can minimize the success rate of pea weevil larvae entry into developing seeds. Greenhouse artificial infestation experiments showed that pea weevil damage in Np genotypes is lower in comparison to wild type genotypes. Therefore, promoting Np formation under field conditions via intercropping can serve as part of an integrated pea weevil management strategy especially for small scale farming systems.

  19. Quarks in Few Body Nuclei

    NASA Astrophysics Data System (ADS)

    Holt, Roy J.

    2016-03-01

    Electron scattering at very high Bjorken x from hadrons provides an excellent test of models, has an important role in high energy physics, and from nuclei, provides a window into short range correlations. Light nuclei have a key role because of the relatively well-known nuclear structure. The development of a novel tritium target for Jefferson Lab has led to renewed interest in the mass three system. For example, deep inelastic scattering experiments in the light nuclei provide a powerful means to determine the neutron structure function. The isospin dependence of electron scattering from mass-3 nuclei provide information on short range correlations in nuclei. The program using the new tritium target will be presented along with a summary of other experiments aimed at revealing the large-x structure of the nucleon.

  20. Nicotinamide attenuates the decrease of astrocytic phosphoprotein PEA-15 in focal cerebral ischemic injury.

    PubMed

    Koh, Phil-Ok

    2012-03-01

    Nicotinamide exerts neuroprotective effects against focal cerebral ischemic injury. Phosphoprotein enriched in astrocytes 15 (PEA-15) is prominently expressed in astrocytes that exert broad anti-apoptotic functions. This study investigated whether nicotinamide modulates PEA-15 and levels of two phosphorylated PEA-15 (Serine 104 and 116) in an animal model of middle cerebral artery occlusion (MCAO)-induced injury. Adult male rats were treated with vehicle or nicotinamide (500 mg/kg) 2 hr after the onset of MCAO and cerebral cortices were collected at 24 hr after MCAO. In a proteomic approach, MCAO induced decreases of PEA-15 levels, while nicotinamide treatment attenuated the injury-induced decrease in PEA-15. The results of Western blot analysis suggest that nicotinamide prevented injury-induced reduction in phospho-PEA-15 (Serine 104) and phospho-PEA-15 (Serine 116) levels. The phosphorylation of PEA-15 exerts anti-apoptotic functions, and reduction of PEA-15 phosphorylation leads to apoptotic cell death. These results suggest that nicotinamide exerts a neuroprotective effect by attenuating the injury-induced decreases of PEA-15 and phospho-PEA-15 (Ser 104 and Ser 116) proteins.

  1. Effect of cooling on Clostridium perfringens in pea soup.

    PubMed

    de Jong, A E I; Rombouts, F M; Beumer, R R

    2004-02-01

    Foods associated with Clostridium perfringens outbreaks are usually abused after cooking. Because of their short generation times, C. perfringens spores and cells can grow out to high levels during improper cooling. Therefore, the potential of C. perfringens to multiply in Dutch pea soup during different cooling times was investigated. Tubes of preheated pea soup (50 degrees C) were inoculated with cocktails of cells or heat-activated spores of this pathogen. The tubes were linearly cooled to 15 degrees C in time spans of 3, 5, 7.5, and 10 h and were subsequently stored in a refrigerator at 3 or 7 degrees C for up to 84 h. Cell numbers increased by 1-log cycle during the 3-h cooling period and reached their maximum after 10 h of cooling. Subsequent refrigeration hardly reduced cell numbers. Cooling of 3.75 liters of pea soup in an open pan showed that this amount of pea soup cooled from 50 to 15 degrees C in 5 h, which will allow a more than 10-fold increase in cell numbers. These findings emphasize the need of good hygienic practices and quick cooling of heated foods after preparation.

  2. 7 CFR 457.140 - Dry pea crop insurance provisions.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... Adequate stand. A population of live plants per acre that will produce at least the yield used to establish... processor/seed company contract, you retain control of the acreage on which the dry peas are grown, you are... not damage due to insufficient or improper application of pest control measures; (d) Plant disease...

  3. 7 CFR 457.140 - Dry pea crop insurance provisions.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Adequate stand. A population of live plants per acre that will produce at least the yield used to establish... processor/seed company contract, you retain control of the acreage on which the dry peas are grown, you are... not damage due to insufficient or improper application of pest control measures; (d) Plant disease...

  4. Faba beans and peas in poultry feed: economic assessment.

    PubMed

    Proskina, Liga; Cerina, Sallija

    2017-10-01

    Broiler diets mainly consist of cereals and protein-rich feed sources; in the EU-27, poultry farming consumes 24% of the total amount of protein-rich feedstuffs. Since the EU produces only 30% of the total quantity of protein crops used for feed, it is necessary to promote the use of traditional European protein crops (beans, peas) for feed in livestock farming. The research aim is to identify economic gains from the production of broiler chicken meat, replacing soybean meal with domestic faba beans and field peas in broiler chicken diets. Adding field peas and faba beans to the broiler feed ration resulted in a significant live weight increase (5.74-11.95%) at the selling age, a decrease in the feed conversion ratio by 0.61-6.06%, and decrease in the product unit cost (15.34-37.06%) as well as an increase in the production efficiency factor (8.70-48.54), compared with the control group. The optimum kind of legume species used in the broiler diet was peas, which were added in the amount of 200 g kg(-1) , resulting in live weight gain, a decrease in the feed conversion ratio and an increase in the production efficiency factor. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  5. A preliminary report on green peas in Alaska

    USDA-ARS?s Scientific Manuscript database

    Western Regional Plant Introduction Station at Pullman, Washington and Seneca Foods Corporation in Dayton, WA collaborated with the Sub-arctic Agricultural Research Unit (SARU) to test a set of established cultivars and breeding lines of green peas under sub-arctic growing conditions. This test set ...

  6. Pea (Pisum sp.) genetic resources, its analysis and exploration

    USDA-ARS?s Scientific Manuscript database

    Pea is important temperate region pulse, with feed, fodder and vegetable uses. Originated and domesticated in Middle East and Mediterranean, it formed important dietary components of early civilizations. Although Pisum is a small genus with two or three species, it is very diverse and structured, r...

  7. Thermostability of sperm nuclei assessed by microinjection into hamster oocytes

    EPA Science Inventory

    Nuclei isolated from spermatozoa of various species (golden hamster, mouse, human, rooster, and the fish tilapia) were heated at 60 degrees-125 degrees C for 20-120 min and then microinjected into hamster oocytes to determine whether they could decondense and develop into pronucl...

  8. Thermostability of sperm nuclei assessed by microinjection into hamster oocytes

    EPA Science Inventory

    Nuclei isolated from spermatozoa of various species (golden hamster, mouse, human, rooster, and the fish tilapia) were heated at 60 degrees-125 degrees C for 20-120 min and then microinjected into hamster oocytes to determine whether they could decondense and develop into pronucl...

  9. Parity violation in nuclei

    SciTech Connect

    Robertson, R.G.H.

    1980-01-01

    A summary of parity violating effects in nuclei is given. Thanks to vigorous experimental and theoretical effort, it now appears that a reasonably well-defined value for the weak isovector ..pi..-nucleon coupling constant can be obtained. There is one major uncertainty in the analysis, namely the M2/E1 mixing ratio for the 2.79 MeV transition in /sup 21/Ne. This quantity is virtually impossible to calculate reliably and must be measured. If it turns out to be much larger than 1, then a null result in /sup 21/Ne is expected no matter what the weak interaction, so an experimental determination is urgently needed. The most promising approach is perhaps a measurement of the pair internal conversion coefficient. Of course, a direct measurement of a pure isovector case is highly desirable, and it is to be hoped that the four ..delta..T = 1 experiments will be pushed still further, and that improved calculations will be made for the /sup 6/Li case. Nuclear parity violation seems to be rapidly approaching an interesting and useful synthesis.

  10. The Metabolism of Quinate in Pea Roots (Purification and Partial Characterization of a Quinate Hydrolyase).

    PubMed Central

    Leuschner, C.; Herrmann, K. M.; Schultz, G.

    1995-01-01

    A quinate (QA) hydrolyase was isolated from pea (Pisum sativum L.) roots. The enzyme converts QA into shikimate by elimination of water. The enzymatic reaction is independent of cofactors and divalent cations. The QA hydrolyase was purified about 1,600-fold to apparent electrophoretic homogeneity in three steps, including bovine serum albumin-affinity chromatography. The enzyme forms oligomers and/or complexes with bovine serum albumin and ovalbumin. The monomer molecular weight of the enzyme is about 15,000. The hydrolyase shows regular Michaelis-Menten kinetics with a Km, of 2.0 mM for QA. Compartmentation studies reveal that the QA hydrolyase is localized in plastids. The QA hydrolyase may function in channeling imported QA into the shikimate pathway to support aromatic amino acid biosynthesis in plastids. PMID:12228477

  11. The Effect of Salinity on the Malic Dehydrogenase of Pea Roots 1

    PubMed Central

    Hason-Porath, Edna; Poljakoff-Mayber, Alexandra

    1969-01-01

    Effect of salinity on malate dehydrogenase activity was studied. Pea root tips contain 2 different malate dehydrogenases. One is located in the particulate, mitochondrial fraction, the other in the soluble, cytoplasmic fraction. Both can act when coupled with either NAD or NADP. Growing plants in Na2SO4 salinated medium did not affect the pattern of the malate dehydrogenases in the root tips. Growing plants in NaCl salinated media resulted in the appearance of a new, third isoenzyme. This new isoenzyme was located in the cytoplasmic fraction. Salinity of both types, when present in growth medium, induced increases in the NADP coupled activity of the mitochondrial malate dehydrogenase. The NAD coupled activity, however, was depressed except in the cytoplasmic fraction of plants grown in media salinated with NaCl to 1 atmosphere. Addition of either of the salts to assay media of enzymes, isolated from plants grown in non salinated substrate, did not have any significant effect. PMID:16657152

  12. Gluon density in nuclei

    SciTech Connect

    Ayala, A.L.; Ducati, M.B.G.; Levin, E.M.

    1996-10-01

    In this talk we present our detailed study (theory and numbers) on the shadowing corrections to the gluon structure functions for nuclei. Starting from rather controversial information on the nucleon structure function which is originated by the recent HERA data, we develop the Glauber approach for the gluon density in a nucleus based on Mueller formula and estimate the value of the shadowing corrections in this case. Then we calculate the first corrections to the Glauber approach and show that these corrections are big. Based on this practical observation we suggest the new evolution equation which takes into account the shadowing corrections and solve it. We hope to convince you that the new evolution equation gives a good theoretical tool to treat the shadowing corrections for the gluons density in a nucleus and, therefore, it is able to provide the theoretically reliable initial conditions for the time evolution of the nucleus-nucleus cascade. The initial conditions should be fixed both theoretically and phenomenologically before to attack such complicated problems as the mixture of hard and soft processes in nucleus-nucleus interactions at high energy or the theoretically reliable approach to hadron or/and parton cascades for high energy nucleus-nucleus interaction. 35 refs., 24 figs., 1 tab.

  13. Non-random chromosome arrangement in triploid endosperm nuclei.

    PubMed

    Baroux, Célia; Pecinka, Ales; Fuchs, Jörg; Kreth, Gregor; Schubert, Ingo; Grossniklaus, Ueli

    2017-02-01

    The endosperm is at the center of successful seed formation in flowering plants. Being itself a product of fertilization, it is devoted to nourish the developing embryo and typically possesses a triploid genome consisting of two maternal and one paternal genome complement. Interestingly, endosperm development is controlled by epigenetic mechanisms conferring parent-of-origin-dependent effects that influence seed development. In the model plant Arabidopsis thaliana, we have previously described an endosperm-specific heterochromatin fraction, which increases with higher maternal, but not paternal, genome dosage. Here, we report a detailed analysis of chromosomal arrangement and association frequency in endosperm nuclei. We found that centromeric FISH signals in isolated nuclei show a planar alignment that may results from a semi-rigid, connective structure between chromosomes. Importantly, we found frequent pairwise association of centromeres, chromosomal segments, and entire arms of chromosomes in 3C endosperm nuclei. These associations deviate from random expectations predicted by numerical simulations. Therefore, we suggest a non-random chromosomal organization in the triploid nuclei of Arabidopsis endosperm. This contrasts with the prevailing random arrangement of chromosome territories in somatic nuclei. Based on observations on a series of nuclei with varying parental genome ratios, we propose a model where chromosomes associate pairwise involving one maternal and one paternal complement. The functional implications of this predicted chromosomal arrangement are discussed.

  14. Genotypic and symbiotic diversity of Rhizobium populations associated with cultivated lentil and pea in sub-humid and semi-arid regions of Eastern Algeria.

    PubMed

    Riah, Nassira; Béna, Gilles; Djekoun, Abdelhamid; Heulin, Karine; de Lajudie, Philippe; Laguerre, Gisèle

    2014-07-01

    The genetic structure of rhizobia nodulating pea and lentil in Algeria, Northern Africa was determined. A total of 237 isolates were obtained from root nodules collected on lentil (Lens culinaris), proteaginous and forage pea (Pisum sativum) growing in two eco-climatic zones, sub-humid and semi-arid, in Eastern Algeria. They were characterised by PCR-restriction fragment length polymorphism (RFLP) of the 16S-23S rRNA intergenic region (IGS), and the nodD-F symbiotic region. The combination of these haplotypes allowed the isolates to be clustered into 26 distinct genotypes, and all isolates were classified as Rhizobium leguminosarum. Symbiotic marker variation (nodD-F) was low but with the predominance of one nod haplotype (g), which had been recovered previously at a high frequency in Europe. Sequence analysis of the IGS further confirmed its high variability in the studied strains. An AMOVA analysis showed highly significant differentiation in the IGS haplotype distribution between populations from both eco-climatic zones. This differentiation was reflected by differences in dominant genotype frequencies. Conversely, no host plant effect was detected. The nodD gene sequence-based phylogeny suggested that symbiotic gene diversity in pea and lentil nodulating rhizobial populations in Algeria was low compared to that reported elsewhere in the world. Copyright © 2014 Elsevier GmbH. All rights reserved.

  15. Mammalian nuclei become licensed for DNA replication during late telophase.

    PubMed

    Dimitrova, Daniela S; Prokhorova, Tatyana A; Blow, J Julian; Todorov, Ivan T; Gilbert, David M

    2002-01-01

    Mcm 2-7 are essential replication proteins that bind to chromatin in mammalian nuclei during late telophase. Here, we have investigated the relationship between Mcm binding, licensing of chromatin for replication, and specification of the dihydrofolate reductase (DHFR) replication origin. Approximately 20% of total Mcm3 protein was bound to chromatin in Chinese hamster ovary (CHO) cells during telophase, while an additional 25% bound gradually and cumulatively throughout G1-phase. To investigate the functional significance of this binding, nuclei prepared from CHO cells synchronized at various times after metaphase were introduced into Xenopus egg extracts, which were either immunodepleted of Mcm proteins or supplemented with geminin, an inhibitor of the Mcm-loading protein Cdt1. Within 1 hour after metaphase, coincident with completion of nuclear envelope formation, CHO nuclei were fully competent to replicate in both of these licensing-defective extracts. However, sites of initiation of replication in each of these extracts were found to be dispersed throughout the DHFR locus within nuclei isolated between 1 to 5 hours after metaphase, but became focused to the DHFR origin within nuclei isolated after 5 hours post-metaphase. Importantly, introduction of permeabilized post-ODP, but not pre-ODP, CHO nuclei into licensing-deficient Xenopus egg extracts resulted in the preservation of a significant degree of DHFR origin specificity, implying that the previously documented lack of specific origin selection in permeabilized nuclei is at least partially due to the licensing of new initiation sites by proteins in the Xenopus egg extracts. We conclude that the functional association of Mcm proteins with chromatin (i.e. replication licensing) in CHO cells takes place during telophase, several hours prior to the specification of replication origins at the DHFR locus.

  16. Local analogues of high-redshift star-forming galaxies: integral field spectroscopy of green peas

    NASA Astrophysics Data System (ADS)

    Lofthouse, E. K.; Houghton, R. C. W.; Kaviraj, S.

    2017-10-01

    We use integral field spectroscopy, from the SWIFT and PALM3K instruments, to perform a spatially resolved spectroscopic analysis of four nearby highly star-forming 'green pea' (GP) galaxies, that are likely analogues of high-redshift star-forming systems. By studying emission-line maps in H α, [N II] λλ6548,6584 and [S II] λλ6716,6731, we explore the kinematic morphology of these systems and constrain properties such as gas-phase metallicities, electron densities and gas-ionization mechanisms. Two of our GPs are rotationally supported while the others are dispersion-dominated systems. The rotationally supported galaxies both show evidence for recent or ongoing mergers. However, given that these systems have intact discs, these interactions are likely to have low-mass ratios (i.e. minor mergers), suggesting that the minor-merger process may be partly responsible for the high star formation rates seen in these GPs. Nevertheless, the fact that the other two GPs appear morphologically undisturbed suggests that mergers (including minor mergers) are not necessary for driving the high star formation rates in such galaxies. We show that the GPs are metal-poor systems (25-40 per cent of solar) and that the gas ionization is not driven by active galactic nuclei (AGN) in any of our systems, indicating that the AGN activity is not coeval with star formation in these starbursting galaxies.

  17. The Pea light-independent photomorphogenesis1 Mutant Results from Partial Duplication of COP1 Generating an Internal Promoter and Producing Two Distinct Transcripts

    PubMed Central

    Sullivan, James A.; Gray, John C.

    2000-01-01

    The pea lip1 (light-independent photomorphogenesis1) mutant shows many of the characteristics of light-grown development when grown in continuous darkness. To investigate the identity of LIP1, cDNAs encoding the pea homolog of COP1, a repressor of photomorphogenesis identified in Arabidopsis, were isolated from wild-type and lip1 pea seedlings. lip1 seedlings contained a wild-type COP1 transcript as well as a larger COP1′ transcript that contained an internal in-frame duplication of 894 bp. The COP1′ transcript segregated with the lip1 phenotype in F2 seedlings and could be translated in vitro to produce a protein of ∼100 kD. The COP1 gene in lip1 peas contained a 7.5-kb duplication, consisting of exons 1 to 7 of the wild-type sequence, located 2.5 kb upstream of a region of genomic DNA identical to the wild-type COP1 DNA sequence. Transcription and splicing of the mutant COP1 gene was predicted to produce the COP1′ transcript, whereas transcription from an internal promoter in the 2.5-kb region of DNA located between the duplicated regions of COP1 would produce the wild-type COP1 transcript. The presence of small quantities of wild-type COP1 transcripts may reduce the severity of the phenotype produced by the mutated COP1′ protein. The genomic DNA sequences of the COP1 gene from wild-type and lip1 peas and the cDNA sequences of COP1 and COP1′ transcripts have been submitted to the EMBL database under the EMBL accession numbers AJ276591, AJ276592, AJ289773, and AJ289774, respectively. PMID:11041887

  18. Molecular Cloning and Evidence for Osmoregulation of the Δ1-Pyrroline-5-Carboxylate Reductase (proC) Gene in Pea (Pisum sativum L.) 12

    PubMed Central

    Williamson, Cynthia L.; Slocum, Robert D.

    1992-01-01

    Several cDNA clones encoding Δ1-pyrroline-5-carboxylate reductase (P5CR, l-proline:NAD[P]+ 5-oxidoreductase, EC 1.5.1.2), which catalyzes the terminal step in proline biosynthesis, were isolated from a pea leaf library screened with a 32P-labeled Aval fragment of a soybean nodule P5CR cDNA (A.J. Delauney, D.P.S. Verma [1990] Mol Gen Genet 221: 299-305). DNA sequence analysis of one full-length 1.3-kb clone (pPPS3) indicated that the pea P5CR gene contains a single major open reading frame encoding a polypeptide of 28,242 Da. Genomic analysis suggested that two to three copies of the P5CR gene are present per haploid genome in pea. The primary structure of pea P5CR is 85% identical with that of soybean and exhibits significant homology to human, yeast, and Escherichia coli P5CR. The sequence of one of four highly conserved domains found in all prokaryotic and eukaryotic P5CRs is similar to the consensus sequence for the NAD(P)H-binding site of other enzymes. The pea P5CR cDNA hybridized to two transcripts, 1.3 and 1.1 kb in size, in polyadenylated RNA purified from leaf tissues of mature, light-grown plants (4 weeks old). Only the 1.3-kb transcript was detected in younger (1 week old) greened seedlings or in etiolated seedlings. In greened seedlings, steady-state levels of this 1.3-kb mRNA increased approximately 5-fold in root tissues within 6 h after plants were irrigated with 0.4 m NaCl, suggesting that expression of the P5CR gene is osmoregulated. Images Figure 3 Figure 4 Figure 5 PMID:11537868

  19. Gravistimulation changes expression of genes encoding putative carrier proteins of auxin polar transport in etiolated pea epicotyls

    NASA Astrophysics Data System (ADS)

    Hoshino, T.; Hitotsubashi, R.; Miyamoto, K.; Tanimoto, E.; Ueda, J.

    STS-95 space experiment has showed that auxin polar transport in etiolated epicotyls of pea (Pisum sativum L. cv. Alaska) seedlings is controlled by gravistimulation. In Arabidopsis thaliana auxin polar transport has considered to be regulated by efflux and influx carrier proteins in plasma membranes, AtPIN1 and AtAUX1, respectively. In order to know how gravistimuli control auxin polar transport in etiolated pea epicotyls at molecular levels, strenuous efforts have been made, resulting in successful isolation of full-length cDNAs of a putative auxin efflux and influx carriers, PsPIN2 and PsAUX1, respectively. Significantly high levels in homology were found on nucleotide and deduced amino acid sequences among PsPIN2, PsPIN1 (accession no. AY222857, Chawla and DeMason, 2003) and AtPINs, and also among PsAUX1, AtAUX1 and their related genes. Phylogenetic analyses based on the deduced amino acid sequences revealed that PsPIN2 belonged to a subclade including AtPIN3, AtPIN4 relating to lateral transport of auxin, while PsPIN1 belonged to the same clade as AtPIN1 relating to auxin polar transport. In the present study, we examined the effects of gravistimuli on the expression of PsPINs and PsAUX1 in etiolated pea seedlings by northern blot analysis. Expression of PsPIN1, PsPIN2 and PsAUX1 in hook region of 3.5-d-old etiolated pea seedlings grown under simulated microgravity conditions on a 3-D clinostat increased as compared with that of the seedlings grown under 1 g conditions. On the other hand, that of PsPIN1 and PsAUX1 in the 1st internode region under simulated microgravity conditions on a 3-D clinostat also increased, while that of PsPIN2 was affected little. These results suggest that expression of PsPIN1, PsPIN2 and PsAUX1 regulating polar/lateral transport of auxin is substantially under the control of gravity. A possible role of PsPINs and PsAUX1 of auxin polar transport in etiolated pea seedlings will also be discussed.

  20. Stem cell mechanics: Auxetic nuclei

    NASA Astrophysics Data System (ADS)

    Wang, Ning

    2014-06-01

    The nuclei of naive mouse embryonic stem cells that are transitioning towards differentiation expand when the cells are stretched and contract when they are compressed. What drives this auxetic phenotype is, however, unclear.

  1. Generalized parton distributions in nuclei

    SciTech Connect

    Vadim Guzey

    2009-12-01

    Generalized parton distributions (GPDs) of nuclei describe the distribution of quarks and gluons in nuclei probed in hard exclusive reactions, such as e.g. deeply virtual Compton scattering (DVCS). Nuclear GPDs and nuclear DVCS allow us to study new aspects of many traditional nuclear effects (nuclear shadowing, EMC effect, medium modifications of the bound nucleons) as well as to access novel nuclear effects. In my talk, I review recent theoretical progress in the area of nuclear GPDs.

  2. Exotic Orbital Modes in Nuclei

    NASA Astrophysics Data System (ADS)

    von Neumann-Cosel, P.

    2003-06-01

    Experimental evidence for two types of collective excitations in nuclei generated by orbital motion is discussed, viz. a magnetic quadrupole twist mode observed in 180° electron scattering experiments and a toroidal electric dipole mode. The latter may be a source of low-energy pygmy dipole resonances observed in many nuclei. This is discussed in detail for the example of 208Pb based on the recent finding of a resonance at particle threshold in a high-resolution (γ, γ') experiment.

  3. The nature of comet nuclei

    NASA Technical Reports Server (NTRS)

    Sykes, Mark V.; Walker, Russell G.

    1992-01-01

    The icy-conglomerate model of comet nuclei has dominated all others since its introduction. It provided a basis for understanding the non-gravitational motions of comets which had perplexed dynamicists up to that time, and provided a focus for understanding cometary composition and origin. The image of comets as dirty snowballs was quickly adopted. Comet nuclei including their trail mass loss rates and refractory to volatile mass ratios are described.

  4. Immunolocalization of pectic polysaccharides during abscission in pea seeds (Pisum sativum L.) and in abscission less def pea mutant seeds.

    PubMed

    Lee, YeonKyeong; Ayeh, Kwadwo Owusu; Ambrose, Mike; Hvoslef-Eide, Anne Kathrine

    2016-08-31

    In pea seeds (Pisum sativum L.), the presence of the Def locus determines abscission event between its funicle and the seed coat. Cell wall remodeling is a necessary condition for abscission of pea seed. The changes in cell wall components in wild type (WT) pea seed with Def loci showing seed abscission and in abscission less def mutant peas were studied to identify the factors determining abscission and non-abscission event. Changes in pectic polysaccharides components were investigated in WT and def mutant pea seeds using immunolabeling techniques. Pectic monoclonal antibodies (1 → 4)-β-D-galactan (LM5), (1 → 5)-α-L-arabinan(LM6), partially de-methyl esterified homogalacturonan (HG) (JIM5) and methyl esterified HG (JIM7) were used for this study. Prior to abscission zone (AZ) development, galactan and arabinan reduced in the predestined AZ of the pea seed and disappeared during the abscission process. The AZ cells had partially de-methyl esterified HG while other areas had highly methyl esterified HG. A strong JIM5 labeling in the def mutant may be related to cell wall rigidity in the mature def mutants. In addition, the appearance of pectic epitopes in two F3 populations resulting from cross between WT and def mutant parents was studied. As a result, we identified that homozygous dominant lines (Def/Def) showing abscission and homozygous recessive lines (def/def) showing non-abscission had similar immunolabeling pattern to their parents. However, the heterogeneous lines (Def/def) showed various immunolabeling pattern and the segregation pattern of the Def locus. Through the study of the complexity and variability of pectins in plant cell walls as well as understanding the segregation patterns of the Def locus using immunolabeling techniques, we conclude that cell wall remodeling occurs in the abscission process and de-methyl esterification may play a role in the non-abscission event in def mutant. Overall, this study contributes new insights into

  5. Programmed cell death in plants: effect of protein synthesis inhibitors and structural changes in pea guard cells.

    PubMed

    Dzyubinskaya, E V; Kiselevsky, D B; Bakeeva, L E; Samuilov, V D

    2006-04-01

    Pea leaf epidermis incubated with cyanide displayed ultrastructural changes in guard cells that are typical of apoptosis. Cycloheximide, an inhibitor of cytoplasmic protein synthesis, and lincomycin, an inhibitor of protein synthesis in chloroplasts and mitochondria, produced different effects on the dynamics of programmed death of guard cells. According to light microscopy data, cycloheximide reinforced and lincomycin suppressed the CN(-)-induced destruction of cell nuclei. Lincomycin lowered the effect of cycloheximide in the light and prevented it in the dark. According to electron microscopy data, the most pronounced effects of cycloheximide in the presence of cyanide were autophagy and a lack of apoptotic condensation of nuclear chromatin, the prevention of chloroplast envelope rupturing and its invagination inside the stroma, and the appearance of particular compartments with granular inclusions in mitochondria. Lincomycin inhibited the CN(-)-induced ultrastructural changes in guard cell nuclei. The data show that programmed death of guard cells may have a combined scenario involving both apoptosis and autophagy and may depend on the action of both cytoplasm synthesized and chloroplast and mitochondrion synthesized proteins.

  6. Phosphoprotein enriched in astrocytes (PEA)-15: A potential therapeutic target in multiple disease states

    PubMed Central

    Greig, Fiona H.; Nixon, Graeme F.

    2014-01-01

    Phosphoprotein enriched in astrocytes-15 (PEA-15) is a cytoplasmic protein that sits at an important junction in intracellular signalling and can regulate diverse cellular processes, such as proliferation and apoptosis, dependent upon stimulation. Regulation of these processes occurs by virtue of the unique interaction of PEA-15 with other signalling proteins. PEA-15 acts as a cytoplasmic tether for the mitogen-activated protein kinases, extracellular signal-regulated kinase 1/2 (ERK1/2) preventing nuclear localisation. In order to release ERK1/2, PEA-15 requires to be phosphorylated via several potential pathways. PEA-15 (and its phosphorylation state) therefore regulates many ERK1/2-dependent processes, including proliferation, via regulating ERK1/2 nuclear translocation. In addition, PEA-15 contains a death effector domain (DED) which allows interaction with other DED-containing proteins. PEA-15 can bind the DED-containing apoptotic adaptor molecule, Fas-associated death domain protein (FADD) which is also dependent on the phosphorylation status of PEA-15. PEA-15 binding of FADD can inhibit apoptosis as bound FADD cannot participate in the assembly of apoptotic signalling complexes. Through these protein–protein interactions, PEA-15-regulated cellular effects have now been investigated in a number of disease-related studies. Changes in PEA-15 expression and regulation have been observed in diabetes mellitus, cancer, neurological disorders and the cardiovascular system. These changes have been suggested to contribute to the pathology related to each of these disease states. As such, new therapeutic targets based around PEA-15 and its associated interactions are now being uncovered and could provide novel avenues for treatment strategies in multiple diseases. PMID:24657708

  7. Pulsed electro-acoustic (PEA) measurements of embedded charge distributions

    NASA Astrophysics Data System (ADS)

    Dennison, J. R.; Pearson, Lee H.

    2013-09-01

    Knowledge of the spatial distribution and evolution of embedded charge in thin dielectric materials has important applications in semiconductor, high-power electronic device, high-voltage DC power cable insulation, high-energy and plasma physics apparatus, and spacecraft industries. Knowing how, where, and how much charge accumulates and how it redistributes and dissipates can predict destructive charging effects. Pulsed Electro-acoustic (PEA) measurements— and two closely related methods, Pressure Wave Propagation (PWP) and Laser Intensity Modulation (LIMM)— nondestructively probe such internal charge distributions. We review the instrumentation, methods, theory and signal processing of simple PEA experiments, as well as the related PPW and LIMM methods. We emphasize system improvements required to achieve high spatial resolution for in vacuo measurements of thin dielectrics charged using electron beam injection.

  8. Purification and Characterization of Pea Chloroplastic Phosphoriboisomerase 12

    PubMed Central

    Skrukrud, Cynthia L.; Gordon, Ilana M.; Dorwin, Sally; Yuan, Xiao-Hua; Johansson, Göte; Anderson, Louise E.

    1991-01-01

    Pea (Pisum sativum L.) chloroplastic phosphoriboisomerase (EC 5.3.1.6) can be purified to apparent homogeneity in less than 2 days time with a 53% yield. Important steps in the purification include heat treatment and pseudoaffinity chromatography on Red H-3BN Sepharose. The purified isomerase has a subunit molecular mass of 26.4 kD. The N-terminal sequence has been determined through 34 residues. pH optima are 7.8 (ribose-5-phosphate) and 7.7 (ribulose-5-phosphate); Km values are 0.9 millimolar (ribose-5-phosphate) and 0.6 millimolar (ribulose-5-phosphate). The enzyme is inhibited by erythrose-4-phosphate, sedoheptulosebisphosphate, glyceraldehyde-3-phosphate, and 3-phosphoglycerate at concentrations close to those found in photosynthesizing chloroplasts. Countercurrent phase partitioning experiments indicate that the pea chloroplastic phosphoriboisomerase interacts physically with phosphoribulokinase. ImagesFigure 1 PMID:16668459

  9. Cutinase is not required for fungal pathogenicity on pea.

    PubMed Central

    Stahl, D J; Schäfer, W

    1992-01-01

    Cutinase, a fungal extracellular esterase, has been proposed to be crucial in the early events of plant infection by many pathogenic fungi. To test the long-standing hypothesis that cutinase of Nectria haematococca (Fusarium solani f sp pisi) is essential to pathogenicity, we constructed cutinase-deficient mutants by transformation-mediated gene disruption of the single cutinase gene of a highly virulent N. haematococca strain. Four independent mutants were obtained lacking a functional cutinase gene, as confirmed by gel blot analyses and enzyme assays. Bioassays of the cutinase-deficient strains showed no difference in pathogenicity and virulence on pea compared to the wild type and a control transformant. We conclude that the cutinase of N. haematococca is not essential for the infection of pea. PMID:1392588

  10. Clinorotation affects mesophyll photosynthetic cells in leaves of pea seedlings.

    PubMed

    Adamchuk, N I

    1998-07-01

    Experiments with autotrophs in altered gravity condition have a grate significant for development of space biology. The main results of investigation in the photosynthetic apparatus state under microgravity condition have based on the experiments with maturity plants and their differentiated cells. The structural and functional organization of photosynthetic cells in seedlings is poor understandable still. Along with chloroplasts preserving a native membrane system in palisade parenchyma cells of the 29-day pea plant leaves in microgravity, chloroplasts with fribly packed or damaged granae, whose thylakoids appeared as vesicles with an electrontransparent content, were also observed. The investigation of preceding process induced these effects have a sense. That is why, the goal of our experiments was to perform the study of a structural organization of the photosynthetic cells of 3-d pair of pea seedlings leaves under the influence of clinorotation.

  11. Analysis of PEA photopolymers at zero spatial frequency limit

    NASA Astrophysics Data System (ADS)

    Gallego, Sergi; Márquez, Andrés; Riquelme, Marina; Neipp, Cristian; Ortuño, Manuel; Beléndez, Augusto; Pascual, Inmaculada

    2012-06-01

    The PEA photopolymer is composed of dipentaerythritol penta/hexa-acrylate as monomer and binder, N-vinyl pirrolidone as crosslinker, ethyl eosin as dye and N-methyl diethanolamine as radical generator. This photopolymer is suitable to work with dispersed liquid crystal molecules in dynamic holographic and diffractive applications. In order to characterize these materials we have analyzed the behaviour of different compositions at zero spatial frequency limit. This method is based on an interferometer that has been successfully applied in the phase-shift versus applied voltage characterization of liquid-crystal displays, in addition to that it has been applied to characterize PVA/AA and PVA/NaAO photopolymers. In PEA case there is no shrinkage since the photopolymer is coverplated. Samples have a glass substrate as the cover plate. In our analysis we have studied the importance of the monomer, crosslinker and crystal liquid molecules concentrations, in the phase shift produced in the layer during photopolymerization process.

  12. Quantitative and Qualitative Involvement of P3N-PIPO in Overcoming Recessive Resistance against Clover Yellow Vein Virus in Pea Carrying the cyv1 Gene

    PubMed Central

    Choi, Sun Hee; Hagiwara-Komoda, Yuka; Atsumi, Go; Shimada, Ryoko; Hisa, Yusuke; Naito, Satoshi

    2013-01-01

    In pea carrying cyv1, a recessive gene for resistance to Clover yellow vein virus (ClYVV), ClYVV isolate Cl-no30 was restricted to the initially infected cells, whereas isolate 90-1 Br2 overcame this resistance. We mapped the region responsible for breaking of cyv1-mediated resistance by examining infection of cyv1 pea with chimeric viruses constructed from parts of Cl-no30 and 90-1 Br2. The breaking of resistance was attributed to the P3 cistron, which is known to produce two proteins: P3, from the main open reading frame (ORF), and P3N-PIPO, which has the N-terminal part of P3 fused to amino acids encoded by a small open reading frame (ORF) called PIPO in the +2 reading frame. We introduced point mutations that were synonymous with respect to the P3 protein but nonsynonymous with respect to the P3N-PIPO protein, and vice versa, into the chimeric viruses. Infection of plants with these mutant viruses revealed that both P3 and P3N-PIPO were involved in overcoming cyv1-mediated resistance. Moreover, P3N-PIPO quantitatively affected the virulence of Cl-no30 in cyv1 pea. Additional expression in trans of the P3N-PIPO derived from Cl-no30, using White clover mosaic virus as a vector, enabled Cl-no30 to move to systemic leaves in cyv1 pea. Susceptible pea plants infected with chimeric ClYVV possessing the P3 cistron of 90-1 Br2, and which were therefore virulent toward cyv1 pea, accumulated more P3N-PIPO than did those infected with Cl-no30, suggesting that the higher level of P3N-PIPO in infected cells contributed to the breaking of resistance by 90-1 Br2. This is the first report showing that P3N-PIPO is a virulence determinant in plants resistant to a potyvirus. PMID:23616656

  13. Quantitative and qualitative involvement of P3N-PIPO in overcoming recessive resistance against Clover yellow vein virus in pea carrying the cyv1 gene.

    PubMed

    Choi, Sun Hee; Hagiwara-Komoda, Yuka; Nakahara, Kenji S; Atsumi, Go; Shimada, Ryoko; Hisa, Yusuke; Naito, Satoshi; Uyeda, Ichiro

    2013-07-01

    In pea carrying cyv1, a recessive gene for resistance to Clover yellow vein virus (ClYVV), ClYVV isolate Cl-no30 was restricted to the initially infected cells, whereas isolate 90-1 Br2 overcame this resistance. We mapped the region responsible for breaking of cyv1-mediated resistance by examining infection of cyv1 pea with chimeric viruses constructed from parts of Cl-no30 and 90-1 Br2. The breaking of resistance was attributed to the P3 cistron, which is known to produce two proteins: P3, from the main open reading frame (ORF), and P3N-PIPO, which has the N-terminal part of P3 fused to amino acids encoded by a small open reading frame (ORF) called PIPO in the +2 reading frame. We introduced point mutations that were synonymous with respect to the P3 protein but nonsynonymous with respect to the P3N-PIPO protein, and vice versa, into the chimeric viruses. Infection of plants with these mutant viruses revealed that both P3 and P3N-PIPO were involved in overcoming cyv1-mediated resistance. Moreover, P3N-PIPO quantitatively affected the virulence of Cl-no30 in cyv1 pea. Additional expression in trans of the P3N-PIPO derived from Cl-no30, using White clover mosaic virus as a vector, enabled Cl-no30 to move to systemic leaves in cyv1 pea. Susceptible pea plants infected with chimeric ClYVV possessing the P3 cistron of 90-1 Br2, and which were therefore virulent toward cyv1 pea, accumulated more P3N-PIPO than did those infected with Cl-no30, suggesting that the higher level of P3N-PIPO in infected cells contributed to the breaking of resistance by 90-1 Br2. This is the first report showing that P3N-PIPO is a virulence determinant in plants resistant to a potyvirus.

  14. PED/PEA-15 controls fibroblast motility and wound closure by ERK1/2-dependent mechanisms.

    PubMed

    Buonomo, Roberta; Giacco, Ferdinando; Vasaturo, Angela; Caserta, Sergio; Guido, Stefano; Pagliara, Valentina; Garbi, Corrado; Mansueto, Gelsomina; Cassese, Angela; Perruolo, Giuseppe; Oriente, Francesco; Miele, Claudia; Beguinot, Francesco; Formisano, Pietro

    2012-05-01

    Cell migration is dependent on the control of signaling events that play significant roles in creating contractile force and in contributing to wound closure. We evaluated wound closure in fibroblasts from mice overexpressing (TgPED) or lacking ped/pea-15 (KO), a gene overexpressed in patients with type 2 diabetes. Cultured skin fibroblasts isolated from TgPED mice showed a significant reduction in the ability to recolonize wounded area during scratch assay, compared to control fibroblasts. This difference was observed both in the absence and in the presence of mytomicin C, an inhibitor of mitosis. In time-lapse experiments, TgPED fibroblasts displayed about twofold lower velocity and diffusion coefficient, as compared to controls. These changes were accompanied by reduced spreading and decreased formation of stress fibers and focal adhesion plaques. At the molecular level, TgPED fibroblasts displayed decreased RhoA activation and increased abundance of phosphorylated extracellular signal-regulated kinase 1/2 (ERK1/2). Inhibition of ERK1/2 activity by PD98059 restored RhoA activation, cytoskeleton organization and cell motility, and almost completely rescued wound closure of TgPED fibroblasts. Interestingly, skin fibroblasts isolated from KO mice displayed an increased wound closure ability. In vivo, healing of dorsal wounds was delayed in TgPED and accelerated in KO mice. Thus, PED/PEA-15 may affect fibroblast motility by a mechanism, at least in part, mediated by ERK1/2.

  15. Identification of Erwinia amylovora, the Fireblight Pathogen, by Colony Hybridization with DNA from Plasmid pEA29

    PubMed Central

    Falkenstein, Hildegard; Bellemann, Peter; Walter, Sabine; Zeller, Wolfgang; Geider, Klaus

    1988-01-01

    All strains of Erwinia amylovora characterized carry a medium-size plasmid of 29 kilobases (pEA29). We mapped this plasmid with various restriction enzymes, cloned the whole DNA into an Escherichia coli plasmid, and subcloned restriction fragments. These DNA species were used for identification of E. amylovora after handling of strains in the laboratory and also in field isolates. About 70 strains of E. amylovora and 24 strains from nine other species, mainly found in plant habitats, were checked in a colony hybridization test. Virulent and avirulent E. amylovora strains reacted positively, whereas the other species were negative. Apart from the hybridization assay, the positive strains were additionally tested for ooze production on rich agar with 5% sucrose and on immature-pear slices. Unspecific background hybridization of non-E. amylovora strains found for hybridization with the whole E. amylovora plasmid was almost eliminated when a 5-kilobase SalI fragment from pEA29 was used as a probe and when the washes after the hybridization procedure were done with high stringency. Under these conditions, E. amylovora could be readily identified from field isolates. Images PMID:16347778

  16. Chloroplast and cytoplasmic enzymes : V. Pea-leaf carbonic anhydrases.

    PubMed

    Kachru, R B; Anderson, L E

    1974-09-01

    Chloroplastic and cytoplasmic forms of pea (Pisum sativum L.) leaf carbonic anhydrase were separated by isoelectric focusing. The two forms have identical pH optima, 7.0 for the hydration reaction and 7.5 for the dehydration reaction, and identical Michaelis constants for CO2, 0.03 M. Neither isozyme is affected by any of several compounds involved in carbon metabolism in the green plant.

  17. [Preparation of flour processed with pigeon pea seed].

    PubMed

    Mueses, C; de León, L; Matute, J; Bressani, R

    1993-03-01

    Pigeon pea is a legume grain of good production capacity and of a relatively high nutritive value, which has not been used in Latin America on the basis of the potential it offers. In this study experiments were conducted to learn about the possibility of processing pigeon pea to yield an intermediate flour with good functional characteristics for food product development. The intermediate pigeon pea flour was produced through a selection of a process to efficiently dehull the grain followed by a thermic process to improve its functional properties and nutritive value. The best dehulling process was subjecting the grain to a vapor treatment for five minutes, followed by a 2-hour dehydration of surface moisture with air at 60 degrees and dehulling with an 8-disc dehuller for 10 minutes. Yield was 84% with 70.7% dehulling efficiency. Pigeon pea flours were prepared by three thermic processes: pressure cooking at 15 lb (121 degrees C) for 5 and 10 minutes as a reference product; cooking and drying with a drum dryer at 120 degrees C and 4 rpm and by extrusion-cooking with the material with 18 and 21% moisture at 270 and 300 degrees F, respectively. Process selection was based on the functional properties such as water absorption index, water solubility index, soluble nitrogen and viscosity, through chemical analysis of protein, available lysine and methionine and residual trypsin inhibitors, and through a biological evaluation of protein digestibility and quality. Both pressure cooking products had similar functional and chemical characteristics, however, the 5-minute cooked product has higher protein quality than the 10-minute product.(ABSTRACT TRUNCATED AT 250 WORDS)

  18. Role of H₂O₂ in pea seed germination.

    PubMed

    Barba-Espín, Gregorio; Hernández, José Antonio; Diaz-Vivancos, Pedro

    2012-02-01

    The imbibition of pea seeds with hydrogen peroxide H₂O₂ increased the germination as well as the seedling growth, producing an invigoration of the seeds. We propose that H₂O₂ could acts as signaling molecule in the beginning of seed germination involving specific changes at proteomic, transcriptomic and hormonal levels. These findings have practical implication in the context of seed priming technologies to invigorate low vigour seeds.

  19. Close-up view Pea pods in Russian Lada greenhouse

    NASA Image and Video Library

    2003-05-12

    ISS007-E-05295 (May 2003) --- Inside the Russian Lada greenhouse, these peas have dried and “gone to seed.” They are part of an experiment to investigate plant development and genetics. The crew of the International Space Station (ISS) will soon harvest the seeds. Eventually, some will be re-planted onboard the ISS and some will be returned to Earth for further study.

  20. Amygdala nuclei critical for emotional learning exhibit unique gene expression patterns.

    PubMed

    Partin, Alexander C; Hosek, Matthew P; Luong, Jonathan A; Lella, Srihari K; Sharma, Sachein A R; Ploski, Jonathan E

    2013-09-01

    The amygdala is a heterogeneous, medial temporal lobe structure that has been implicated in the formation, expression and extinction of emotional memories. This structure is composed of numerous nuclei that vary in cytoarchitectonics and neural connections. In particular the lateral nucleus of the amygdala (LA), central nucleus of the amygdala (CeA), and the basal (B) nucleus contribute an essential role to emotional learning. However, to date it is still unclear to what extent these nuclei differ at the molecular level. Therefore we have performed whole genome gene expression analysis on these nuclei to gain a better understanding of the molecular differences and similarities among these nuclei. Specifically the LA, CeA and B nuclei were laser microdissected from the rat brain, and total RNA was isolated from these nuclei and subjected to RNA amplification. Amplified RNA was analyzed by whole genome microarray analysis which revealed that 129 genes are differentially expressed among these nuclei. Notably gene expression patterns differed between the CeA nucleus and the LA and B nuclei. However gene expression differences were not considerably different between the LA and B nuclei. Secondary confirmation of numerous genes was performed by in situ hybridization to validate the microarray findings, which also revealed that for many genes, expression differences among these nuclei were consistent with the embryological origins of these nuclei. Knowing the stable gene expression differences among these nuclei will provide novel avenues of investigation into how these nuclei contribute to emotional arousal and emotional learning, and potentially offer new genetic targets to manipulate emotional learning and memory.

  1. Amygdala nuclei critical for emotional learning exhibit unique gene expression patterns ✩

    PubMed Central

    Partin, Alexander C.; Hosek, Matthew P.; Luong, Jonathan A.; Lella, Srihari K.; Sharma, Sachein A.R.; Ploski, Jonathan E.

    2014-01-01

    The amygdala is a heterogeneous, medial temporal lobe structure that has been implicated in the formation, expression and extinction of emotional memories. This structure is composed of numerous nuclei that vary in cytoarchitectonics and neural connections. In particular the Lateral nucleus of the Amygdala (LA), Central nucleus of the Amygdala (CeA), and the Basal (B) nucleus contribute an essential role to emotional learning. However, to date it is still unclear to what extent these nuclei differ at the molecular level. Therefore we have performed whole genome gene expression analysis on these nuclei to gain a better understanding of the molecular differences and similarities among these nuclei. Specifically the LA, CeA and B nuclei were laser microdissected from the rat brain, and total RNA was isolated from these nuclei and subjected to RNA amplification. Amplified RNA was analyzed by whole genome microarray analysis which revealed that 129 genes are differentially expressed among these nuclei. Notably gene expression patterns differed between the CeA nucleus and the LA and B nuclei. However gene expression differences were not considerably different between the LA and B nuclei. Secondary confirmation of numerous genes was performed by in situ hybridization to validate the microarray findings, which also revealed that for many genes, expression differences among these nuclei were consistent with the embryological origins of these nuclei. Knowing the stable gene expression differences among these nuclei will provide novel avenues of investigation into how these nuclei contribute to emotional arousal and emotional learning, and potentially offer new genetic targets to manipulate emotional learning and memory. PMID:23831498

  2. Hyperglycemia aggravates decreases of PEA-15 and its two phosphorylated forms in cerebral ischemia

    PubMed Central

    SUNG, Jin-Hee; KOH, Phil-Ok

    2017-01-01

    Diabetes is a metabolic health disorder and an important risk factor for stroke. Phosphoprotein enriched in astrocytes 15 (PEA-15) is a multifunctional protein modulating cell proliferation, survival, apoptosis and glucose metabolism. This study investigated whether diabetes modulates the expression of PEA-15 and two phosphorylated forms (Ser 104 and Ser 116) in middle cerebral artery occlusion (MCAO)-induced brain injury. Male Sprague-Dawley rats were administrated with streptozotocin (40 mg/kg) and were underwent right middle cerebral artery occlusion (MCAO) 4 weeks after streptozotocin injection. Brain tissues were collected 24 hr after MCAO and stained using triphenyltetrazolium chloride. Western blot analysis was performed to elucidate the expression of PEA-15 and two phosphorylated forms (Ser 104 and Ser 116) in right cerebral cortex. Infarct volume during MCAO injury was severely increased in diabetic animals compared to non-diabetic animals. We identified the decrease in PEA-15 in animals that underwent MCAO using proteomic approach. PEA-15 expression during MCAO was strongly decreased in diabetic animals compared to non-diabetic animals. Western blots analysis confirmed that diabetes exacerbated the decrease in PEA-15 expression after MCAO. Moreover, decrease in expression of phospho-PEA-15 (Ser 104 and Ser 116) was greater in diabetic than in non-diabetic animals. These results suggested that a diabetic condition may aggravate brain damage through decreasing expression of PEA-15 and phospho-PEA-15 (Ser 104 and Ser 116) in ischemic brain injury. PMID:28216548

  3. Final Step of Phosphatidic Acid Synthesis in Pea Chloroplasts Occurs in the Inner Envelope Membrane 1

    PubMed Central

    Andrews, Jaen; Ohlrogge, John B.; Keegstra, Kenneth

    1985-01-01

    The second enzyme of phosphatidic acid synthesis from glycerol-3-phosphate, 1-acylglycerophospate acyltransferase, was localized to the inner envelope membrane of pea chloroplasts. The activity of this enzyme was measured by both a coupled enzyme assay and a direct enzyme assay. Using the coupled enzyme assay, phosphatidic acid phosphatase was also localized to the inner envelope membrane, although this enzyme has very low activity in pea chloroplasts. The addition of UDP-galactose to unfractionated pea chloroplast envelope preparations did not result in significant conversion of newly synthesized diacylglycerol to monogalactosyldiacylglycerol. Thus, the envelope synthesized phosphatidic acid may not be involved in galactolipid synthesis in pea chloroplasts. PMID:16664266

  4. Addition of sucralose enhances the release of satiety hormones in combination with pea protein.

    PubMed

    Geraedts, Maartje C P; Troost, Freddy J; Saris, Wim H M

    2012-03-01

    Exposing the intestine to proteins or tastants, particularly sweet, affects satiety hormone release. There are indications that each sweetener has different effects on this release, and that combining sweeteners with other nutrients might exert synergistic effects on hormone release. STC-1 cells were incubated with acesulfame-K, aspartame, saccharine, sucralose, sucrose, pea, and pea with each sweetener. After a 2-h incubation period, cholecystokinin(CCK) and glucagon-like peptide 1 (GLP-1) concentrations were measured. Using Ussing chamber technology, the mucosal side of human duodenal biopsies was exposed to sucrose, sucralose, pea, and pea with each sweetener. CCK and GLP-1 levels were measured in basolateral secretions. In STC-1 cells, exposure to aspartame, sucralose, sucrose, pea, and pea with sucralose increased CCK levels, whereas GLP-1 levels increased after addition of all test products. Addition of sucrose and sucralose to human duodenal biopsies did not affect CCK and GLP-1 release; addition of pea stimulated CCK and GLP-1 secretion. Combining pea with sucrose and sucralose induced even higher levels of CCK and GLP-1. Synchronous addition of pea and sucralose to enteroendocrine cells induced higher levels of CCK and GLP-1 than addition of each compound alone. This study shows that combinations of dietary compounds synergize to enhance satiety hormone release. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Pre-fractionation strategies to resolve pea (Pisum sativum) sub-proteomes

    PubMed Central

    Meisrimler, Claudia-Nicole; Menckhoff, Ljiljana; Kukavica, Biljana M.; Lüthje, Sabine

    2015-01-01

    Legumes are important crop plants and pea (Pisum sativum L.) has been investigated as a model with respect to several physiological aspects. The sequencing of the pea genome has not been completed. Therefore, proteomic approaches are currently limited. Nevertheless, the increasing numbers of available EST-databases as well as the high homology of the pea and medicago genome (Medicago truncatula Gaertner) allow the successful identification of proteins. Due to the un-sequenced pea genome, pre-fractionation approaches have been used in pea proteomic surveys in the past. Aside from a number of selective proteome studies on crude extracts and the chloroplast, few studies have targeted other components such as the pea secretome, an important sub-proteome of interest due to its role in abiotic and biotic stress processes. The secretome itself can be further divided into different sub-proteomes (plasma membrane, apoplast, cell wall proteins). Cell fractionation in combination with different gel-electrophoresis, chromatography methods and protein identification by mass spectrometry are important partners to gain insight into pea sub-proteomes, post-translational modifications and protein functions. Overall, pea proteomics needs to link numerous existing physiological and biochemical data to gain further insight into adaptation processes, which play important roles in field applications. Future developments and directions in pea proteomics are discussed. PMID:26539198

  6. Relating physico-chemical properties of frozen green peas (Pisum sativum L.) to sensory quality.

    PubMed

    Nleya, Kathleen M; Minnaar, Amanda; de Kock, Henriëtte L

    2014-03-30

    The acceptability of frozen green peas depends on their sensory quality. There is a need to relate physico-chemical parameters to sensory quality. In this research, six brands of frozen green peas representing product sold for retail and caterer's markets were purchased and subjected to descriptive sensory evaluation and physico-chemical analyses (including dry matter content, alcohol insoluble solids content, starch content, °Brix, residual peroxidase activity, size sorting, hardness using texture analysis and colour measurements) to assess and explain product quality. The sensory quality of frozen green peas, particularly texture properties, were well explained using physico-chemical methods of analysis notably alcohol insoluble solids, starch content, hardness and °Brix. Generally, retail class peas were of superior sensory quality to caterer's class peas although one caterer's brand was comparable to the retail brands. Retail class peas were sweeter, smaller, greener, more moist and more tender than the caterer's peas. Retail class peas also had higher °Brix, a(*) , hue and chroma values; lower starch, alcohol insoluble solids, dry matter content and hardness measured. The sensory quality of frozen green peas can be partially predicted by measuring physico-chemical parameters particularly °Brix and to a lesser extent hardness by texture analyser, alcohol insoluble solids, dry matter and starch content. © 2013 Society of Chemical Industry.

  7. [Possibility of using flour of pigeon pea in products prepared with rice or wheat flour].

    PubMed

    Mueses, C; de León, L; Bressani, R

    1993-03-01

    The present study reports on the development of foods containing processed pigeon pea (Cajanus cajan) flour. The pigeon pea flours described in a previous publication were prepared from dehulled pigeon peas by cooking in autoclave, by extrusion-cooking and by cooking/dehydration by drum-drying. Mixtures of cooked pigeon peas and rice were first evaluated biological through a protein complementation design using NPR. The results of this study showed that the two products had high protein quality and were similar when mixed in ratios of 80:20 to 40:60. For the evaluation of the processed pigeon pea flour, mixtures with rice (80:20) were used. All pigeon pea flours gave similar protein quality values. On the basis of these results three products were developed and tested. One was a gruel ("atole"), a second a fruit-flavored thick drink with and without 15% milk. Cookies were also prepared with a series of blends of pigeon pea flour (extrusion-cooked) and wheat. The gruel and the fruit flavored products had high acceptability based on a sensory evaluation test. Cookies with 100% pigeon pea flour were unacceptable, however, mixtures of 75% wheat flour and 25% pigeon pea flour gave cookies of attractive appearance and good taste. The study showed the possibility of preparing and utilizing tropical grain legume flours for food products of relatively high acceptability and nutritive value.

  8. Pre-fractionation strategies to resolve pea (Pisum sativum) sub-proteomes.

    PubMed

    Meisrimler, Claudia-Nicole; Menckhoff, Ljiljana; Kukavica, Biljana M; Lüthje, Sabine

    2015-01-01

    Legumes are important crop plants and pea (Pisum sativum L.) has been investigated as a model with respect to several physiological aspects. The sequencing of the pea genome has not been completed. Therefore, proteomic approaches are currently limited. Nevertheless, the increasing numbers of available EST-databases as well as the high homology of the pea and medicago genome (Medicago truncatula Gaertner) allow the successful identification of proteins. Due to the un-sequenced pea genome, pre-fractionation approaches have been used in pea proteomic surveys in the past. Aside from a number of selective proteome studies on crude extracts and the chloroplast, few studies have targeted other components such as the pea secretome, an important sub-proteome of interest due to its role in abiotic and biotic stress processes. The secretome itself can be further divided into different sub-proteomes (plasma membrane, apoplast, cell wall proteins). Cell fractionation in combination with different gel-electrophoresis, chromatography methods and protein identification by mass spectrometry are important partners to gain insight into pea sub-proteomes, post-translational modifications and protein functions. Overall, pea proteomics needs to link numerous existing physiological and biochemical data to gain further insight into adaptation processes, which play important roles in field applications. Future developments and directions in pea proteomics are discussed.

  9. Mechanism of gibberellin-dependent stem elongation in peas

    NASA Technical Reports Server (NTRS)

    Cosgrove, D. J.; Sovonick-Dunford, S. A.

    1989-01-01

    Stem elongation in peas (Pisum sativum L.) is under partial control by gibberellins, yet the mechanism of such control is uncertain. In this study, we examined the cellular and physical properties that govern stem elongation, to determine how gibberellins influence pea stem growth. Stem elongation of etiolated seedlings was retarded with uniconozol, a gibberellin synthesis inhibitor, and the growth retardation was reversed by exogenous gibberellin. Using the pressure probe and vapor pressure osmometry, we found little effect of uniconozol and gibberellin on cell turgor pressure or osmotic pressure. In contrast, these treatments had major effects on in vivo stress relaxation, measured by turgor relaxation and pressure-block techniques. Uniconozol-treated plants exhibited reduced wall relaxation (both initial rate and total amount). The results show that growth retardation is effected via a reduction in the wall yield coefficient and an increase in the yield threshold. These effects were largely reversed by exogenous gibberellin. When we measured the mechanical characteristics of the wall by stress/strain (Instron) analysis, we found only minor effects of uniconozol and gibberellin on the plastic compliance. This observation indicates that these agents did not alter wall expansion through effects on the mechanical (viscoelastic) properties of the wall. Our results suggest that wall expansion in peas is better viewed as a chemorheological, rather than a viscoelastic, process.

  10. Mechanism of gibberellin-dependent stem elongation in peas

    NASA Technical Reports Server (NTRS)

    Cosgrove, D. J.; Sovonick-Dunford, S. A.

    1989-01-01

    Stem elongation in peas (Pisum sativum L.) is under partial control by gibberellins, yet the mechanism of such control is uncertain. In this study, we examined the cellular and physical properties that govern stem elongation, to determine how gibberellins influence pea stem growth. Stem elongation of etiolated seedlings was retarded with uniconozol, a gibberellin synthesis inhibitor, and the growth retardation was reversed by exogenous gibberellin. Using the pressure probe and vapor pressure osmometry, we found little effect of uniconozol and gibberellin on cell turgor pressure or osmotic pressure. In contrast, these treatments had major effects on in vivo stress relaxation, measured by turgor relaxation and pressure-block techniques. Uniconozol-treated plants exhibited reduced wall relaxation (both initial rate and total amount). The results show that growth retardation is effected via a reduction in the wall yield coefficient and an increase in the yield threshold. These effects were largely reversed by exogenous gibberellin. When we measured the mechanical characteristics of the wall by stress/strain (Instron) analysis, we found only minor effects of uniconozol and gibberellin on the plastic compliance. This observation indicates that these agents did not alter wall expansion through effects on the mechanical (viscoelastic) properties of the wall. Our results suggest that wall expansion in peas is better viewed as a chemorheological, rather than a viscoelastic, process.

  11. Green Pea Galaxies Reveal Secrets of Lyα Escape

    NASA Astrophysics Data System (ADS)

    Yang, Huan; Malhotra, Sangeeta; Gronke, Max; Rhoads, James E.; Dijkstra, Mark; Jaskot, Anne; Zheng, Zhenya; Wang, Junxian

    2016-04-01

    We analyze archival Lyα spectra of 12 “Green Pea” galaxies observed with the Hubble Space Telescope, model their Lyα profiles with radiative transfer models, and explore the dependence of the Lyα escape fraction on various properties. Green Pea galaxies are nearby compact starburst galaxies with [O iii] λ5007 equivalent widths (EWs) of hundreds of Å. All 12 Green Pea galaxies in our sample show Lyα lines in emission, with an Lyα EW distribution similar to high-redshift Lyα emitters. Combining the optical and UV spectra of Green Pea galaxies, we estimate their Lyα escape fractions and find correlations between Lyα escape fraction and kinematic features of Lyα profiles. The escape fraction of Lyα in these galaxies ranges from 1.4% to 67%. We also find that the Lyα escape fraction depends strongly on metallicity and moderately on dust extinction. We compare their high-quality Lyα profiles with single H i shell radiative transfer models and find that the Lyα escape fraction anticorrelates with the derived H i column densities. Single-shell models fit most Lyα profiles well, but not the ones with the highest escape fractions of Lyα. Our results suggest that low H i column density and low metallicity are essential for Lyα escape and make a galaxy an Lyα emitter.

  12. Gravitropic response and circumnutation in pea (Pisum sativum) seedling roots.

    PubMed

    Kim, Hye-jeong; Kobayashi, Akie; Fujii, Nobuharu; Miyazawa, Yutaka; Takahashi, Hideyuki

    2016-05-01

    Plant circumnutation is a helical movement of growing organs such as shoots and roots. Gravitropic response is hypothesized to act as an external oscillator in shoot circumnutation, although this is subject to debate. The relationship between circumnutational movement and gravitropic response in roots remains unknown. In this study, we analyzed circumnutation of agravitropic roots using the ageotropum pea (Pisum sativum) mutant, and compared it with that of wild-type (cv. Alaska) pea roots. We further examined the relationship of gravitropic response to circumnutation of Alaska seedling roots by removing the gravisensing tissue (the root cap) and by treating the roots with auxin transport inhibitors. Alaska roots displayed circumnutational movements with a period of approximately 150 min, whereas ageotropum roots did not exhibit distinct circumnutational movement. Removal of the root cap in Alaska roots reduced gravitropic response and circumnutational movements. Treatment of Alaska roots with auxin transport inhibitors, 2,3,5-triiodobenzoic acid (TIBA) and N-(1-naphthyl)phthalamic acid (NPA), dramatically reduced gravitropic response and circumnutational movements. These results suggest that a gravity-regulated auxin transport is involved in circumnutation of pea seedling roots. © 2015 Scandinavian Plant Physiology Society.

  13. Mechanism of gibberellin-dependent stem elongation in peas.

    PubMed

    Cosgrove, D J; Sovonick-Dunford, S A

    1989-01-01

    Stem elongation in peas (Pisum sativum L.) is under partial control by gibberellins, yet the mechanism of such control is uncertain. In this study, we examined the cellular and physical properties that govern stem elongation, to determine how gibberellins influence pea stem growth. Stem elongation of etiolated seedlings was retarded with uniconozol, a gibberellin synthesis inhibitor, and the growth retardation was reversed by exogenous gibberellin. Using the pressure probe and vapor pressure osmometry, we found little effect of uniconozol and gibberellin on cell turgor pressure or osmotic pressure. In contrast, these treatments had major effects on in vivo stress relaxation, measured by turgor relaxation and pressure-block techniques. Uniconozol-treated plants exhibited reduced wall relaxation (both initial rate and total amount). The results show that growth retardation is effected via a reduction in the wall yield coefficient and an increase in the yield threshold. These effects were largely reversed by exogenous gibberellin. When we measured the mechanical characteristics of the wall by stress/strain (Instron) analysis, we found only minor effects of uniconozol and gibberellin on the plastic compliance. This observation indicates that these agents did not alter wall expansion through effects on the mechanical (viscoelastic) properties of the wall. Our results suggest that wall expansion in peas is better viewed as a chemorheological, rather than a viscoelastic, process.

  14. Cavitation inception from bubble nuclei.

    PubMed

    Mørch, K A

    2015-10-06

    The tensile strength of ordinary water such as tap water or seawater is typically well below 1 bar. It is governed by cavitation nuclei in the water, not by the tensile strength of the water itself, which is extremely high. Different models of the nuclei have been suggested over the years, and experimental investigations of bubbles and cavitation inception have been presented. These results suggest that cavitation nuclei in equilibrium are gaseous voids in the water, stabilized by a skin which allows diffusion balance between gas inside the void and gas in solution in the surrounding liquid. The cavitation nuclei may be free gas bubbles in the bulk of water, or interfacial gaseous voids located on the surface of particles in the water, or on bounding walls. The tensile strength of these nuclei depends not only on the water quality but also on the pressure-time history of the water. A recent model and associated experiments throw new light on the effects of transient pressures on the tensile strength of water, which may be notably reduced or increased by such pressure changes.

  15. Cavitation inception from bubble nuclei

    PubMed Central

    Mørch, K. A.

    2015-01-01

    The tensile strength of ordinary water such as tap water or seawater is typically well below 1 bar. It is governed by cavitation nuclei in the water, not by the tensile strength of the water itself, which is extremely high. Different models of the nuclei have been suggested over the years, and experimental investigations of bubbles and cavitation inception have been presented. These results suggest that cavitation nuclei in equilibrium are gaseous voids in the water, stabilized by a skin which allows diffusion balance between gas inside the void and gas in solution in the surrounding liquid. The cavitation nuclei may be free gas bubbles in the bulk of water, or interfacial gaseous voids located on the surface of particles in the water, or on bounding walls. The tensile strength of these nuclei depends not only on the water quality but also on the pressure–time history of the water. A recent model and associated experiments throw new light on the effects of transient pressures on the tensile strength of water, which may be notably reduced or increased by such pressure changes. PMID:26442138

  16. Reflection asymmetric shapes in nuclei

    SciTech Connect

    Ahmad, I.; Carpenter, M.P.; Emling, H.; Holzmann, R.; Janssens, R.V.F.; Khoo, T.L.; Moore, E.F.; Morss, L.R.; Durell, J.L.; Fitzgerald, J.B.; Mowbary, A.S.; Hotchkiss, M.A.; Phillips, W.R.; Drigert, M.W.; Ye, D.; Benet, P.; Manchester Univ. . Dept. of Physics; EG and G Idaho, Inc., Idaho Falls, ID; Notre Dame Univ., IN; Purdue Univ., Lafayette, IN )

    1989-01-01

    Experimental data show that there is no even-even nucleus with a reflection asymmetric shape in its ground state. Maximum octupole- octupole correlations occur in nuclei in the mass 224 (N{approximately}134, Z{approximately}88) region. Parity doublets, which are the characteristic signature of octupole deformation, have been observed in several odd mass Ra, Ac and Pa nuclei. Intertwined negative and positive parity levels have been observed in several even-even Ra and Th nuclei above spin {approximately}8{Dirac h}. In both cases, the opposite parity states are connected by fast El transitions. In some medium-mass nuclei intertwined negative and positive parity levels have also been observed above spin {approximately}7{Dirac h}. The nuclei which exhibit octupole deformation in this mass region are {sup 144}Ba, {sup 146}Ba and {sub 146}Ce; {sup 142}Ba, {sup 148}Ce, {sup 150}Ce and {sup 142}Xe do not show these characteristics. No case of parity doublet has been observed in the mass 144 region. 32 refs., 16 figs., 1 tab.

  17. The role of recovery of mitochondrial structure and function in desiccation tolerance of pea seeds.

    PubMed

    Wang, Wei-Qing; Cheng, Hong-Yan; Møller, Ian M; Song, Song-Quan

    2012-01-01

    Mitochondrial repair is of fundamental importance for seed germination. When mature orthodox seeds are imbibed and germinated, they lose their desiccation tolerance in parallel. To gain a better understanding of this process, we studied the recovery of mitochondrial structure and function in pea (Pisum sativum cv. Jizhuang) seeds with different tolerance to desiccation. Mitochondria were isolated and purified from the embryo axes of control and imbibed-dehydrated pea seeds after (re-)imbibition for various times. Recovery of mitochondrial structure and function occurred both in control and imbibed-dehydrated seed embryo axes, but at different rates and to different maximum levels. The integrity of the outer mitochondrial membrane reached 96% in all treatments. However, only the seeds imbibed for 12 h and then dehydrated recovered the integrity of the inner mitochondrial membrane (IMM) and State 3 (respiratory state in which substrate and ADP are present) respiration (with NADH and succinate as substrate) to the control level after re-imbibition. With increasing imbibition time, the degree to which each parameter recovered decreased in parallel with the decrease in desiccation tolerance. The tolerance of imbibed seeds to desiccation increased and decreased when imbibed in CaCl(2) and methylviologen solution, respectively, and the recovery of the IMM integrity similarly improved and weakened in these two treatments, respectively. Survival of seeds after imbibition-dehydration linearly increased with the increase in ability to recover the integrity of IMM and State 3 respiration, which indicates that recovery of mitochondrial structure and function during germination has an important role in seed desiccation tolerance.

  18. Differential Expression of the S-Adenosyl-l-Methionine Synthase Genes during Pea Development1

    PubMed Central

    Gómez-Gómez, Lourdes; Carrasco, Pedro

    1998-01-01

    Two genes coding for S-adenosyl-l-methionine synthase (SAMS, EC 2.5.1.6) were previously isolated from pea (Pisum sativum) ovaries. Both SAMS genes were highly homologous throughout their coding regions but showed a certain degree of sequence divergence within the 5′ and the 3′ untranslated regions. These regions have been used as gene-specific probes to analyze the differential expression of SAMS1 and SAMS2 genes in pea plants. The ribonuclease protection assay revealed different expression patterns for each individual gene. SAMS1 was strongly expressed in nearly all tissues, especially in roots. SAMS2 expression was weaker, reaching its highest level at the apex. Following pollination, SAMS1 was specifically up-regulated, whereas SAMS2 was expressed constitutively. The up-regulation of SAMS1 during ovary development was also observed in unpollinated ovaries treated with auxins. In unpollinated ovaries an increase in SAMS1 expression was observed as a consequence of ethylene production associated with the emasculation process. In senescing ovaries both SAMS1 and SAMS2 genes showed increased expression. Ethylene treatment of unpollinated ovaries led to an increase in the SAMS1 mRNA level. However, SAMS2 expression remained unchangeable after ethylene treatment, indicating that SAMS2 induction during ovary senescence was not ethylene dependent. SAMS mRNAs were localized by in situ hybridization at the endocarp of developing fruits and in the ovules of senescing ovaries. Our results indicate that the transcriptional regulation of SAMS genes is developmentally controlled in a specific way for each gene. PMID:9625692

  19. Enumeration of islets by nuclei counting and light microscopic analysis

    PubMed Central

    Pisania, Anna; Papas, Klearchos K.; Powers, Daryl E.; Rappel, Michael J.; Omer, Abdulkadir; Bonner-Weir, Susan; Weir, Gordon C.; Colton, Clark K.

    2010-01-01

    Islet enumeration in impure preparations by conventional dithizone staining and visual counting is inaccurate and operator dependent. We examined nuclei counting for measuring the total number of cells in islet preparations, and we combined it with morphological analysis by light microscopy (LM) for estimating the volume fraction of islets in impure preparations. Cells and islets were disrupted with lysis solution and shear, and accuracy of counting successively diluted nuclei suspensions was verified with: (1) visual counting in a hemacytometer after staining with crystal violet, and automatic counting by (2) aperture electrical resistance measurement and (3) flow cytometer measurement after staining with 7-aminoactinomycin-D. DNA content averaged 6.5 and 6.9 pg DNA/cell for rat and human islets, respectively, in agreement with literature estimates. With pure rat islet preparations, precision improved with increasing counts, and samples with about 160 or more islets provided a coefficient of variation of about 6%. Aliquots of human islet preparations were processed for LM analysis by stereological point counting. Total nuclei counts and islet volume fraction from LM analysis were combined to obtain the number of islet equivalents (IE). Total number of IE by the standard method of dithizone staining/manual counting was overestimated by about 90% compared to LM/nuclei counting for 12 freshly isolated human islet research preparations. Nuclei counting combined with islet volume fraction measurements from light microscopy is a novel method for achieving accurate islet enumeration. PMID:20697375

  20. A peptide that binds the pea aphid gut impedes entry of Pea enation mosaic virus into the aphid hemocoel

    SciTech Connect

    Liu Sijun; Sivakumar, S.; Sparks, Wendy O.; Miller, W. Allen; Bonning, Bryony C.

    2010-05-25

    Development of ways to block virus transmission by aphids could lead to novel and broad-spectrum means of controlling plant viruses. Viruses in the Luteoviridae enhanced are obligately transmitted by aphids in a persistent manner that requires virion accumulation in the aphid hemocoel. To enter the hemocoel, the virion must bind and traverse the aphid gut epithelium. By screening a phage display library, we identified a 12-residue gut binding peptide (GBP3.1) that binds to the midgut and hindgut of the pea aphid Acyrthosiphon pisum. Binding was confirmed by labeling the aphid gut with a GBP3.1-green fluorescent protein fusion. GBP3.1 reduced uptake of Pea enation mosaic virus (Luteoviridae) from the pea aphid gut into the hemocoel. GBP3.1 also bound to the gut epithelia of the green peach aphid and the soybean aphid. These results suggest a novel strategy for inhibiting plant virus transmission by at least three major aphid pest species.

  1. Nuclear pore ion channel activity in live syncytial nuclei.

    PubMed

    Bustamante, Jose Omar

    2002-05-01

    Nuclear pore complexes (NPCs) are important nanochannels for the control of gene activity and expression. Most of our knowledge of NPC function has been derived from isolated nuclei and permeabilized cells in cell lysates/extracts. Since recent patch-clamp work has challenged the dogma that NPCs are freely permeable to small particles, a preparation of isolated living nuclei in their native liquid environment was sought and found: the syncytial nuclei in the water of the coconut Cocos nucifera. These nuclei have all properties of NPC-mediated macromolecular transport (MMT) and express foreign green fluorescent protein (GFP) plasmids. They display chromatin movement, are created by particle aggregation or by division, can grow by throwing filaments to catch material, etc. This study shows, for the first time, that living NPCs engaged in MMT do not transport physiological ions - a phenomenon that explains observations of nucleocytoplasmic ion gradients. Since coconuts are inexpensive (less than US$1/nut per litre), this robust preparation may contribute to our understanding of NPCs and cell nucleus and to the development of biotechnologies for the production of DNA, RNA and proteins.

  2. Octupole shapes in heavy nuclei

    SciTech Connect

    Ahmad, I.

    1994-08-01

    Theoretical calculations and measurements show the presence of strong octupole correlations in thecyround states and low-lying states of odd-mass and odd-odd nuclei in the RaPa region. Evidence for octupole correlations is provided by the observation of parity doublets and reductions in M1 matrix elements, decoupling parameters, and Coriolis matrix elements Involving high-j states. Enhancement of E1 transition rates has also been observed for some of the octupole deformed nuclei. The most convincing argument for octupole deformation is provided by the similarities of the reduced alpha decay rates to the two members of parity doublets.

  3. Chromatin structure in barley nuclei.

    PubMed

    Mithieux, G; Roux, B

    1983-10-03

    In order to study the chromatin structure of a higher plant we used a high-yield method, which allows one to obtain up to 10(9) nuclei/kg fresh barley leaves. Significant amounts of low-ionic-strength-soluble chromatin can be extracted from these nuclei. Physicochemical properties were examined and discussed. Electric birefringence allowed us to observe the same transition in electro-optical properties as has been observed for animal chromatin, and suggested the existence of a symetrical structure occurring for approximately six nucleosomes. Circular dichroism showed that barley oligonucleosomes exhibit a higher molar ellipticity at 282 nm than total soluble chromatin and than their animal counterparts.

  4. Post-reproductive parthenogenetic pea aphids (Acyrthosiphon pisum) are visually identifiable and disproportionately positioned distally to clonal colonies

    PubMed Central

    Diamond, Julia Daisy; Henneman, Nathaniel Fath; Levitis, Daniel A.

    2016-01-01

    The role of kin-selection in the evolution of post-reproductive life is controversial. While anthropological and demographic studies strongly suggest that humans and a few other species experience kin selection for significant post-reproductive survival, these results are necessarily correlational. Understanding could therefore be advanced by the development of a globally available, field and laboratory tractable experimental model of kin-selected post-reproductive survival. In only one invertebrate (Quadrartus yoshinomiyai, a gall-forming aphid endemic to Japan) have individuals too old to reproduce been shown to be both numerous in natural habitats and able to help close relatives survive or reproduce. Pea aphids, (Acyrthosiphon pisum), common, tractable organisms, frequently outlive their reproductive ages in laboratories, live in tight interacting groups that are often clonal, and therefore should be evaluated as potential model organisms for the study of adaptive post-reproductive life. The first major step in this process is to identify an optimal method for assessing if a parthenogenetic adult is post-reproductive. We evaluated three methods, relying respectively on isolation in clip cages, visual examination for embryonic eyespots, and dissection. In every case each method identified the same individuals as reproductive versus post-reproductive. While the clip-cage method requires a multi-day wait to produce data, and dissection is inevitably fatal, the eyespot method is quick (under one minute per individual) easy, and non-invasive. This method makes it possible to accurately assess the post-reproductive status of a large number of parthenogenetic pea aphids. We demonstrate the usefulness of the eyespot method in showing that while reproductively valuable adults tend to place themselves near the centers of clonal colonies, less valuable post-reproductive adults are more often at or beyond the edges of colonies. These encouraging early results provide both

  5. Post-reproductive parthenogenetic pea aphids (Acyrthosiphon pisum) are visually identifiable and disproportionately positioned distally to clonal colonies.

    PubMed

    Saberski, Erik T; Diamond, Julia Daisy; Henneman, Nathaniel Fath; Levitis, Daniel A

    2016-01-01

    The role of kin-selection in the evolution of post-reproductive life is controversial. While anthropological and demographic studies strongly suggest that humans and a few other species experience kin selection for significant post-reproductive survival, these results are necessarily correlational. Understanding could therefore be advanced by the development of a globally available, field and laboratory tractable experimental model of kin-selected post-reproductive survival. In only one invertebrate (Quadrartus yoshinomiyai, a gall-forming aphid endemic to Japan) have individuals too old to reproduce been shown to be both numerous in natural habitats and able to help close relatives survive or reproduce. Pea aphids, (Acyrthosiphon pisum), common, tractable organisms, frequently outlive their reproductive ages in laboratories, live in tight interacting groups that are often clonal, and therefore should be evaluated as potential model organisms for the study of adaptive post-reproductive life. The first major step in this process is to identify an optimal method for assessing if a parthenogenetic adult is post-reproductive. We evaluated three methods, relying respectively on isolation in clip cages, visual examination for embryonic eyespots, and dissection. In every case each method identified the same individuals as reproductive versus post-reproductive. While the clip-cage method requires a multi-day wait to produce data, and dissection is inevitably fatal, the eyespot method is quick (under one minute per individual) easy, and non-invasive. This method makes it possible to accurately assess the post-reproductive status of a large number of parthenogenetic pea aphids. We demonstrate the usefulness of the eyespot method in showing that while reproductively valuable adults tend to place themselves near the centers of clonal colonies, less valuable post-reproductive adults are more often at or beyond the edges of colonies. These encouraging early results provide both

  6. Antihypertensive Properties of a Pea Protein Hydrolysate during Short- and Long-Term Oral Administration to Spontaneously Hypertensive Rats.

    PubMed

    Girgih, Abraham T; Nwachukwu, Ifeanyi D; Onuh, John O; Malomo, Sunday A; Aluko, Rotimi E

    2016-05-01

    This study investigated short-term (24 h) and long-term (5 wk) systolic blood pressure (SBP)-lowering effects in spontaneously hypertensive rats (SHR) of a 5 kDa membrane pea protein hydrolysate permeate (PPH-5) produced through thermoase hydrolysis of pea protein isolate (PPI). Amino acid analysis showed that the PPH-5 had lower contents of sulfur-containing amino acids than the PPI. Size-exclusion chromatography indicated mainly low molecular weight (<10 kDa) peptides in PPH-5 but not in the PPI. The PPH-5 had renin and angiotensin converting enzyme inhibition IC50 values of 0.57 and 0.10 mg/mL (P < 0.05), respectively, and consisted mainly of peptides with 2 to 6 amino acids. Mass spectrometry analysis revealed mainly hydrophilic tetrapeptide sequences. After a single oral administration (100 mg/kg body weight) to SHR, the unheated PPI showed weakest (P < 0.05) SBP-lowering effect with a -4 mm Hg maximum when compared to -25 mm Hg for heat-treated PPI and -36 mm Hg for PPH-5. Incorporation of the PPH-5 as 0.5% or 1% (w/w) casein substitute in the SHR diet produced maximum SBP reductions of -22 or -26 mm Hg (P < 0.05), respectively after 3 wk. In comparison, the unhydrolyzed PPI produced a maximum SBP reduction of -17 mm Hg also after 3 wk. Potency of the pea products decreased in the 4th and 5th wk, though SBP values of the treated rats were still lower than the untreated control. We conclude that the antihypertensive potency of PPH-5 may have been due to the presence of easily absorbed hydrophilic peptides. © 2016 Institute of Food Technologists®

  7. Biosynthesis of Indoleacetic Acid from Tryptophan-14C in Cell-free Extracts of Pea Shoot Tips 1

    PubMed Central

    Moore, Thomas C.; Shaner, Coralie A.

    1967-01-01

    A 2-step, 1-dimensional thin-layer chromatographic procedure for isolating indoleacetic acid (IAA) was developed and utilized in investigations of the biosynthesis of IAA from tryptophan-14C in cell-free extracts of pea (Pisum sativum L.) shoot tips. Identification of a 14C-product as IAA was by (a) co-chromatography of authentic IAA and 14C-product on thin-layer chromatography, and (b) gas-liquid and thin-layer chromatography of authentic and presumptive IAA methyl esters. Dialysis of enzyme extracts and addition of α-ketoglutaric acid and pyridoxal phosphate to reaction mixtures resulted in approximately 2- to 3-fold increases in net yields of IAA over yields in non-dialyzed reaction mixtures which did not contain additives essential to a transaminase reaction of tryptophan. Addition of thiamine pyrophosphate to reaction mixtures further enhanced net biosynthesis of IAA. It is concluded that the formation of indolepyruvic acid and its subsequent decarboxylation probably are sequential reactions in the major pathway of IAA biosynthesis from tryptophan in cell-free extracts of Pisum shoot tips. Comparison of maximum net IAA biosynthesis in extracts of shoot tips of etiolated and light-grown dwarf and tall pea seedlings revealed an order, on a unit protein N basis, of: light-grown tall > light-grown dwarf > etiolated tall ≅ etiolated dwarf. It is concluded that the different rates of stem elongation among etiolated and light-grown dwarf and tall pea seedlings are correlated, in general, with differences in net IAA biosynthesis and sensitivity of the tissues to IAA. PMID:16656720

  8. Novel cross-linked alcohol-insoluble solid (CL-AIS) affinity gel from pea pod for pectinesterase purification.

    PubMed

    Wu, Ming-Chang; Lin, Guan-Hui; Wang, Yuh-Tai; Jiang, Chii-Ming; Chang, Hung-Min

    2005-10-05

    Alcohol-insoluble solids (AIS) from pea pod were cross-linked (CL-AIS) and used as an affinity gel matrix to isolate pectin esterases (PEs) from tendril shoots of chayote (TSC) and jelly fig achenes (JFA), and the results were compared with those isolated by ion-exchange chromatography with a commercial resin. CL-AIS gel matrix in a column displayed poor absorption and purification fold of PE; however, highly methoxylated CL-AIS (HM-CL-AIS), by exposing CL-AIS to methanolic sulfuric acid to increase the degree of esterification (DE) to 92%, facilitated the enzyme purification. The purified TSC PE and JFA PE by the HM-CL-AIS column were proofed as a single band on an SDS-PAGE gel, showing that the HM-CL-AIS column was a good matrix for purification of PE, either with alkaline isoelectric point (pI) (TSC PE) or with acidic pI (JFA PE).

  9. Effect of succinate on phosphate solubilization in nitrogen fixing bacteria harbouring chick pea and their effect on plant growth.

    PubMed

    Iyer, Bhagya; Rajput, Mahendrapal Singh; Rajkumar, Shalini

    2017-09-01

    Diverse nitrogen fixing bacteria harbouring chick pea rhizosphere and root nodules were tested for multiple plant growth promoting traits like tricalcium phosphate (TCP) and rock phosphate (RP) solubilization, production of ammonia, indole 3-acetic acid, chitinase, phytase and alkaline phosphatase. Isolates belonged to diverse genus like Enterobacter, Acinetobacter, Erwinia, Pseudomonas, Rhizobium, Sinorhizobium, Ensifer, Klebsiella, etc. Most isolates solubilized TCP and RP along with the lowering of media pH, indicating acidification to be the chief mechanism behind this solubilization. However, lowering of media pH and P release decreased by 32-100% when media was supplemented with succinate, a major component of plant root exudates indicating succinate mediated repression of P solubilization. Maximum TCP and RP solubilization with P release of 850μg/mL and 2088μg/mL was obtained with lowering of media pH up to 2.8 and 3.3 for isolate E43 and PSB1 respectively. This pH drop changed to 4.4 and 4.8 with 80% and 87% decrease in P solubilization in the presence of succinate. Maximum 246μg/mL indole 3-acetic acid production in Lh3, 44.8U/mL chitinase activity in MB3, 11.3U/mL phytase activity in I91 and 9.4U/mL alkaline phosphatase activity in SM1 were also obtained. Most isolates showed multiple PGP traits which resulted in significant plant growth promotion of chick pea plants. Present study shows repression of P solubilization by succinate for various bacterial groups which might be one of the reasons why phosphate solubilizing bacteria which perform well in vitro often fail in vivo. Studying this repression mechanism might be critical in understanding the in vivo efficacy. Copyright © 2017. Published by Elsevier GmbH.

  10. International Symposium on Exotic Nuclei

    NASA Astrophysics Data System (ADS)

    Penionzhkevich, Yu. E.; Cherepanov, E. A.

    Methods of production of light exotic nuclei and study of their ptoperties -- Superheavy elements. Syhnthesis and properties -- Nuclear fission -- Nuclear reactions -- rare processes, decay and nuclear structure -- Experimental set-ups and future projects -- Radioactive beams. Production and research programmes -- Public relations.

  11. Proton Distribution in Heavy Nuclei

    DOE R&D Accomplishments Database

    Johnson, M. H; Teller, E.

    1953-11-13

    It is reasoned that, from considerations connected with beta-decay stability and Coulomb repulsion forces, a neutron excess is developed on the surface of heavy nuclei. Several consequences of this qualitative analysis in nucleon interactions are briefly noted. (K.S.)

  12. Chiral electroweak currents in nuclei

    DOE PAGES

    Riska, D. O.; Schiavilla, R.

    2017-01-10

    Here, the development of the chiral dynamics based description of nuclear electroweak currents is reviewed. Gerald E. (Gerry) Brown’s role in basing theoretical nuclear physics on chiral Lagrangians is emphasized. Illustrative examples of the successful description of electroweak observables of light nuclei obtained from chiral effective field theory are presented.

  13. Electromagnetic structure of light nuclei

    SciTech Connect

    Pastore, Saori

    Here, the present understanding of nuclear electromagnetic properties including electromagnetic moments, form factors and transitions in nuclei with A ≤ 10 is reviewed. Emphasis is on calculations based on nuclear Hamiltonians that include two- and three-nucleon realistic potentials, along with one- and two-body electromagnetic currents derived from a chiral effective field theory with pions and nucleons.

  14. Transitional nuclei near shell closures

    SciTech Connect

    Mukherjee, G.

    2014-08-14

    High spin states in Bismuth and Thallium nuclei near the Z = 82 shell closure and Cesium nuclei near the N = 82 shell closure in A = 190 and A = 130 regions, respectively, have been experimentally investigated using heavy-ion fusion evaporation reaction and by detecting the gamma rays using the Indian National Gamma Array (INGA). Interesting shape properties in these transitional nuclei have been observed. The results were compared with the neighboring nuclei in these two regions. The total Routhian surface (TRS) calculations have been performed for a better understanding of the observed properties. In mass region A = 190, a change in shape from spherical to deformed has been observd around neutron number N = 112 for the Bi (Z = 83) isotopes with proton number above the magic gap Z = 82, whereas, the shape of Tl (Z = 81) isotopes with proton number below the magic gap Z = 82 remains stable as a function of neutron number. An important transition from aplanar to planar configuration of angular momentum vectors leading to the occurance of nuclar chirality and magnetic rotation, respectively, has been proposed for the unique parity πh{sub 11/2}⊗νh{sub 11/2} configuration in Cs isotopes in the mass region A ∼ 130 around neutron number N = 79. These results are in commensurate with the TRS calculations.

  15. Electromagnetic structure of light nuclei

    DOE PAGES

    Pastore, Saori

    2016-03-25

    Here, the present understanding of nuclear electromagnetic properties including electromagnetic moments, form factors and transitions in nuclei with A ≤ 10 is reviewed. Emphasis is on calculations based on nuclear Hamiltonians that include two- and three-nucleon realistic potentials, along with one- and two-body electromagnetic currents derived from a chiral effective field theory with pions and nucleons.

  16. Octupole correlation effects in nuclei

    SciTech Connect

    Chasman, R.R.

    1992-01-01

    Octupole correlation effects in nuclei are discussed from the point of view of many-body wavefunctions as well as mean-field methods. The light actinides, where octupole effects are largest, are considered in detail. Comparisons of theory and experiment are made for energy splittings of parity doublets; E1 transition matrix elements and one-nucleon transfer reactions.

  17. Octupole correlation effects in nuclei

    SciTech Connect

    Chasman, R.R.

    1992-08-01

    Octupole correlation effects in nuclei are discussed from the point of view of many-body wavefunctions as well as mean-field methods. The light actinides, where octupole effects are largest, are considered in detail. Comparisons of theory and experiment are made for energy splittings of parity doublets; E1 transition matrix elements and one-nucleon transfer reactions.

  18. Understanding nuclei: progress and challenges

    SciTech Connect

    Dean, D. J.

    2008-04-17

    Nuclear theory today aims for a comprehensive theoretical framework that can describe all nuclei. I discuss recent progress in this pursuit and the associated challenges as we move forward, paying particular attention to progress in the applications of coupled-cluster theory to the challenges.

  19. Nuclei and propeller cavitation inception

    SciTech Connect

    Gindroz, B.; Billet, M.L.

    1994-12-31

    Propeller cavitation inception tests were conducted in the Grand Tunnel Hydrodynamique (GTH) of the Bassin d`Essaid des Carenes. Both acoustic and visual cavitation inception were determined for leading-edge sheet, travelling bubble, and tip vortex. These data were obtained for specific water quality conditions. The water quality was determined from cavitation susceptibility meter measurements for degassed water (maximum liquid tension, few nuclei), low injection rate of microbubbles (medium liquid tension, low nuclei concentration), medium injection rate of microbubbles (medium liquid tension, high nuclei concentration) and high injection rate of microbubbles (minimum liquid tension, high nuclei concentration). Results clearly demonstrate a different influence of water quality for each type of cavitation. Little variation in cavitation inception index for a significant increase in liquid tension and microbubble size distribution was found for leading-edge sheet; however, tip vortex cavitation inception index decreased significantly for an increase in liquid tension. In addition, a dependency on event rate was determined for tip vortex cavitation inception.

  20. Identification of tolerance to Fusarium root rot in wild pea germplasm with high levels of partial resistance

    USDA-ARS?s Scientific Manuscript database

    Fusarium root rot, caused by Fusarium solani f. sp. pisi, is a serious root rot pathogen affecting peas in all pea growing areas of the USA and is damaging in both dryland and irrigated pea fields. Partial resistance to Fusarium root rot in 44 accessions from the Pisum Core Collection located in Pu...

  1. Characterization of regenerated butterfly pea (Clitoria ternatea L.) accessions for morphological, phenology, reproductive and potential nutraceutical, pharmaceutical trait utilization.

    USDA-ARS?s Scientific Manuscript database

    Butterfly pea, Clitoria ternatea, has been used in Africa as a companion crop and in the United States as an ornamental. The USDA, ARS, PGRCU curates 28 butterfly pea accessions. Butterfly pea accessions were transplanted from about 30-day-old seedlings to the field in Griffin, GA, around 01 June ...

  2. EFFORT TOWARDS A WORLD PEA (Pisum sativum L.) GERMPLASM CORE COLLECTION: The case for common markers and data compatibility

    USDA-ARS?s Scientific Manuscript database

    Using SSR and RBIP markers, several major world pea germplasm collections have been analyzed and core collections formed. These include over 1,200 pea accessions of Chinese origin contained within a larger set of over 2,000 accessions analyzed by 21 SSR loci , 310 of 5,394 USDA pea germplasm acce...

  3. Association mapping of agronomic and quality traits in USDA pea single-plant collection

    USDA-ARS?s Scientific Manuscript database

    Association mapping is an efficient approach for the identification of the molecular basis of agronomic traits in crop plants. For this purpose in pea (Pisum sativum L.), we genotyped and phenotyped individual lines of the single-plant derived core collection of the USDA pea single-plant (PSP) colle...

  4. Plasma palmitoylethanolamide (PEA) as a potential biomarker for impaired coronary function.

    PubMed

    Quercioli, Alessandra; Carbone, Federico; Bonaventura, Aldo; Liberale, Luca; Pataky, Zoltan; Thomas, Aurélien; Lenglet, Sébastien; Lauer, Estelle; Golay, Alain; Dallegri, Franco; Di Marzo, Vincenzo; Schindler, Thomas H; Montecucco, Fabrizio

    2017-03-15

    Among endocannabinoid (EC)-related mediators, Oleoyl-ethanolamide (OEA) and Palmitoyl-ethanolamide (PEA), two endogenous PPARα agonists with lipolytic and anti-inflammatory action, respectively, are being actively investigated. Here, we assessed the potential association between plasma levels of PEA and OEA and coronary function in a cohort including normal, overweight, obese, and morbidly obese (MOB) individuals. Myocardial perfusion and endothelium-related myocardial blood flow (MBF) responses to cold pressor test (CPT) and during pharmacological vasodilation with dipyridamole were measured with (13)N-ammonia positron emission tomography/computed tomography. OEA and PEA were extracted from human plasma by liquid-liquid extraction, separated by liquid chromatography and quantified by mass spectrometry. Serum levels of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 (VCAM-1) were measured by colorimetric enzyme-linked immunosorbent assay. Circulating levels of PEA and VCAM-1 were increased in MOB as compared to normal weight subjects. Circulating levels of OEA and PEA were associated with body mass index, but not with adhesion molecules. Increases of PEA levels were associated with and predictive of worsened coronary function in MOB and the overall cohort studied. Plasma levels of PEA are increased in MOB patients and associated with coronary dysfunction as a functional precursor of CAD process. Larger trials are needed to confirm PEA as a potential circulating biomarker of coronary dysfunction in both MOB patients and the general population. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  5. Release of pea germplasm with Fusarium resistance combined with desirable yield and anti-lodging traits

    USDA-ARS?s Scientific Manuscript database

    Fusarium root rot caused by Fusarium solani f. sp. pisi (Fsp) and Fusarium wilt caused by Fusarium oxysporum f. sp. pisi (Fop) races 1, 2 and 5, negatively impact the pea industry worldwide. Limited pea germplasm with agronomically acceptable characteristics combined with resistance to these disease...

  6. Screening of pea genotypes for resistance to root rot caused by Rhizoctonia solani AG 8, 2012.

    USDA-ARS?s Scientific Manuscript database

    Rhizoctonia solani AG 8 is one of the major pathogens that causes pea root rot and stunting in the Columbia Basin of Oregon and Washington. The disease is most severe in fields where wheat has been mono-cropped for a number of years or where cereal cover crops are incorporated just before pea seedin...

  7. Estimating demand for perennial pigeon pea in Malawi using choice experiments.

    PubMed

    Waldman, Kurt B; Ortega, David L; Richardson, Robert B; Snapp, Sieglinde S

    2017-01-01

    Perennial crops have numerous ecological and agronomic advantages over their annual counterparts. We estimate discrete choice models to evaluate farmers' preferences for perennial attributes of pigeon pea intercropped with maize in central and southern Malawi. Pigeon pea is a nitrogen-fixing leguminous crop, which has the potential to ameliorate soil fertility problems related to continuous maize cultivation, which are common in Southern Africa. Adoption of annual pigeon pea is relatively low but perennial production of pigeon pea may be more appealing to farmers due to some of the ancillary benefits associated with perenniality. We model perennial production of pigeon pea as a function of the attributes that differ between annual and perennial production: lower labor and seed requirements resulting from a single planting with multiple harvests, enhanced soil fertility and higher levels of biomass production. The primary tradeoff associated with perennial pigeon pea intercropped with maize is competition with maize in subsequent years of production. While maize yield is approximately twice as valuable to farmers as pigeon pea yield, we find positive yet heterogeneous demand for perenniality driven by soil fertility improvements and pigeon pea grain yield.

  8. PeaT1-induced systemic acquired resistance in tobacco follows salicylic acid-dependent pathway.

    PubMed

    Zhang, Wei; Yang, Xiufen; Qiu, Dewen; Guo, Lihua; Zeng, Hongmei; Mao, Jianjun; Gao, Qiufeng

    2011-04-01

    Systemic acquired resistance (SAR) is an inducible defense mechanism which plays a central role in protecting plants from pathogen attack. A new elicitor, PeaT1 from Alternaria tenuissima, was expressed in Escherichia coil and characterized with systemic acquired resistance to tobacco mosaic virus (TMV). PeaT1-treated plants exhibited enhanced systemic resistance with a significant reduction in number and size of TMV lesions on wild tobacco leaves as compared with control. The quantitative analysis of TMV CP gene expression with real-time quantitative PCR showed there was reduction in TMV virus concentration after PeaT1 treatment. Similarly, peroxidase (POD) activity and lignin increased significantly after PeaT1 treatment. The real-time quantitative PCR revealed that PeaT1 also induced the systemic accumulation of pathogenesis-related gene, PR-1a and PR-1b which are the markers of systemic acquired resistance (SAR), NPR1 gene for salicylic acid (SA) signal transduction pathway and PAL gene for SA synthesis. The accumulation of SA and the failure in development of similar level of resistance as in wild type tobacco plants in PeaT1 treated nahG transgenic tobacco plants indicated that PeaT1-induced resistance depended on SA accumulation. The present work suggested that the molecular mechanism of PeaT1 inducing disease resistance in tobacco was likely through the systemic acquired resistance pathway mediated by salicylic acid and the NPR1 gene.

  9. Grass pea and neurolathyrism: farmers' perception on its consumption and protective measure in North Shewa, Ethiopia.

    PubMed

    Girma, Anteneh; Tefera, Beneberu; Dadi, Legesse

    2011-03-01

    Neurolathyrism in Ethiopia is caused by food dependency on grass pea (Lathyrus sativus L.). In the study area, a large proportion of the farmers are growing grass pea since it can withstand harsh environments. Socio-economic factors (poverty; lack of money to buy other food legumes) and environmental problems (such as water logging and frost hazards) influence consumption of grass pea. Most of the respondents have the idea that some chemical contained in grass pea causes a health problem. Different processing and preparation methods are used to prepare grass pea into different food forms. The major processing methods include washing and soaking, as the farmers apply these methods mainly because they assume that the chemical that causes lathyrism, scientifically known as β-ODAP (β-N-oxalyl-L-α,β-diaminopropionic acid) is reduced through washing and soaking. The farmers adopt different strategies to avoid the problem of lathyrism such as avoiding consumption of grass pea in the form that they suspect to cause the problem, blending/mixing with other crops, applying different processing/detoxification methods. Since grass pea is consumed with a fear of lathyrism, future research should concentrate either on developing grass pea varieties with safe level of β-ODAP content or improving the traditional/indigenous processing methods.

  10. Effects of Planting Date on Biomass Production by Grass pea (Lathyrus sativus L.).

    USDA-ARS?s Scientific Manuscript database

    Grass pea is a cool-season legume commonly grown for human consumption and livestock feed in Asia and East Africa. A better understanding of the agronomic importance of planting date and the influence of photoperiod may lead to improved management strategies for cultivation of grass pea in the sout...

  11. Ersiphe trifolii-a newly recognized powdery mildew pathogen of pea.

    USDA-ARS?s Scientific Manuscript database

    Population diversity of powdery mildews infecting pea (Pisum sativum) in the US Pacific Northwest was investigated in order to assess inconsistent resistance performances of pea genotypes in different environments. Phylogenetic analyses based on ITS sequences, in combination with assessment of morph...

  12. 2015 Outbreak of Cyclosporiasis Linked to the Consumption of Imported Sugar Snap Peas in Ontario, Canada.

    PubMed

    Whitfield, Yvonne; Johnson, Karen; Hanson, Heather; Huneault, Danya

    2017-09-06

    An outbreak of cyclosporiasis in Ontario, Canada, was investigated in the fall of 2015. Thirty-five confirmed and 10 probable cases were linked to the investigation. Epidemiological and food safety evidence implicated fresh sugar snap peas imported from Guatemala as the source of the outbreak. We describe here the first documented cyclosporiasis outbreak in Canada involving the consumption of sugar snap peas.

  13. A positive role for PEA3 in HER2-mediated breast tumour progression

    PubMed Central

    Myers, E; Hill, A D K; Kelly, G; McDermott, E W; O'Higgins, N J; Young, L S

    2006-01-01

    Overexpression of HER2 is associated with an adverse prognosis in breast cancer. Despite this, the mechanism of its transcriptional regulation remains poorly understood. PEA3, a MAP kinase (MAPK)-activated member of the Ets transcription factor family has been implicated in the transcriptional regulation of HER2. The direction of its modulation remains controversial. We assessed relative levels of PEA3 expression and DNA binding in primary breast cultures derived from patient tumours (n=18) in the presence of an activated MAPK pathway using Western blotting and shift analysis. Expression of PEA3 in breast tumours from patients of known HER2 status (n=107) was examined by immunohistochemistry. In primary breast cancer cell cultures, growth factors induced interaction between PEA3 and its DNA response element. Upregulation of PEA3 expression in the presence of growth factors associated with HER2 positivity and axillary lymph node metastasis (P=0.034 and 0.049, respectively). PEA3 expression in breast cancer tissue associated with reduced disease-free survival (P<0.001), Grade III tumours (P<0.0001) and axillary lymph node metastasis (P=0.026). Co-expression of PEA3 and HER2 significantly associated with rate of recurrence compared to patients who expressed HER2 alone (P=0.0039). These data support a positive role for PEA3 in HER2-mediated oncogenesis in breast cancer. PMID:17060941

  14. Genetic Diversity of Chinese and Global Pea (Pisum sativum L.) Collections.

    USDA-ARS?s Scientific Manuscript database

    Pea (Pisum sativum L.) is an important food and feed legume grown across many temperate regions of the world, especially from Asia to Europe and North America. The goal of this study was to use 30 informative pea microsatellite markers to compare genetic diversity in a global core from the USDA and ...

  15. Conformational distortion of α-phenylethyl amine in cryogenic matrices - a matrix isolation VCD study.

    PubMed

    Pollok, Corina H; Merten, Christian

    2016-05-21

    The chiral amine α-phenylethyl amine (PEA) was isolated in cryogenic matrices and investigated using vibrational circular dichroism (VCD) and infrared spectroscopy. The potential energy surface (PES) of PEA features five different conformers connected by relatively low conformational transition states. Based on the IR spectra, it could be confirmed that all conformational energy barriers are passed at a deposition temperature of 20 K, and that only the global minimum conformation of PEA is populated in both argon and nitrogen matrices. However, differences in the calculated and experimental VCD spectra indicate deviations from the minimum structure by perturbation of the phenyl ring as well as of the amine orientation. The degree of the perturbation is found to also depend on the choice of the host gas, which shows the subtle influence of the environment on the conformational distortion of PEA.

  16. Phosphorylation is the switch that turns PEA-15 from tumor suppressor to tumor promoter

    PubMed Central

    Sulzmaier, Florian J.; Opoku-Ansah, John; Ramos, Joe W.

    2012-01-01

    Abnormal ERK signaling is implicated in many human diseases including cancer. This signaling cascade is a good target for the therapy of certain malignancies because of its important role in regulating cell proliferation and survival. The small phosphoprotein PEA-15 is a potent regulator of the ERK signaling cascade, and, by acting on this pathway, has been described to have both tumor-suppressor and tumor-promoter functions. However, the exact mechanism by which PEA-15 drives the outcome one way or the other remains unclear. We propose that the cellular environment is crucial in determining PEA-15 protein function by affecting the protein’s phosphorylation state. We hypothesize that only unphosphorylated PEA-15 can act as a tumor-suppressor and that phosphorylation alters the interaction with binding partners to promote tumor development. In order to use PEA-15 as a prognostic marker or therapeutic target it is therefore important to evaluate its phosphorylation status. PMID:22694972

  17. Formation of electric dipoles in pea stem tissue due to an electric field

    NASA Astrophysics Data System (ADS)

    Ahmadi, Fatemeh; Farahani, Elham

    2016-07-01

    For examining the effect of an electrical field (DC) on pea seed, we exposed the pea seeds to electric fields with intensities 1, 4 and 7 kV/cm for 30, 230, 430 and 630 seconds. The tests were repeated three times, and each iteration had 5 seeds. Then, the seeds were moved to packaged plates. Finally, microscopic observation of the pea stem tissue showed that the application of a DC electrical field caused a deformation in the pea stem tissue. The results led us to examine the deformation of the tissue theoretically and to address that deformation as an electrostatic problem. In this regard, we modeled the pea stem based on the formation of electric dipoles. Then, theoretically, we calculated the force acting on each xylem section by coding, and the results were consistent with the experimental data.

  18. Magnetic moments of neutron deficient yttrium nuclei

    SciTech Connect

    Berks; El Hajjaji, O.; Fahad, M.; Hassani, R.; Giroux, J.; Marest, G.; Marguier, G.; Stone, N.J.; Rikovska, J.; Green, V.R.; and others

    1987-12-10

    This paper describes recent low temperature nulcear orientation (LTNO) work on neutron deficient /sup 85m,86,86m/Y nuclei. Results are compared with experimental systematics of neighbouring nuclei and particle core coupling calculations.

  19. Bean α-amylase inhibitor 1 in transgenic peas (Pisum sativum) provides complete protection from pea weevil (Bruchus pisorum) under field conditions

    PubMed Central

    Morton, Roger L.; Schroeder, Hart E.; Bateman, Kaye S.; Chrispeels, Maarten J.; Armstrong, Eric; Higgins, Thomas J. V.

    2000-01-01

    Two α-amylase inhibitors, called αAI-1 and αAI-2, that share 78% amino acid sequence identity and have a differential specificity toward mammalian and insect α-amylases are present in different accessions of the common bean (Phaseolus vulgaris). Using greenhouse-grown transgenic peas (Pisum sativum), we have shown previously that expression of αAI-1 in pea seeds can provide complete protection against the pea weevil (Bruchus pisorum). Here, we report that αAI-1 also protects peas from the weevil under field conditions. The high degree of protection is explained by our finding that αAI-1 inhibits pea bruchid α-amylase by 80% over a broad pH range (pH 4.5–6.5). αAI-2, on the other hand, is a much less effective inhibitor of pea bruchid α-amylase, inhibiting the enzyme by only 40%, and only in the pH 4.0–4.5 range. Nevertheless, this inhibitor was still partially effective in protecting field-grown transgenic peas against pea weevils. The primary effect of αAI-2 appeared to be a delay in the maturation of the larvae. This contrasts with the effect of αAI-1, which results in larval mortality at the first or second instar. These results are discussed in relationship to the use of amylase inhibitors with different specificities to bring about protection of crops from their insect pests or to decrease insect pest populations below the economic injury level. PMID:10759552

  20. Studies of Heavy-Ion Reactions and Transuranic Nuclei

    SciTech Connect

    Schroeder, W. Udo

    2016-07-28

    Studies of heavy-ion reactions and transuranic nuclei performed by the University of Rochester Nuclear Science Research Group have been successful in furthering experimental systematics and theoretical understanding of the behavior of nuclear systems excited to their limits of stability. The theoretical results explain specifically the “boiling” and “vaporization” of atomic nuclei, but are more generally applicable to isolated, quantal many-particle systems which, under thermal or mechanical stresses, all disintegrate by evaporation, via surface cluster emission, or via fission-like processes. Accompanying experimental investigations by the group have demonstrated several new types of dynamical instability of nuclei: In central, “head-on” collisions, target nuclei exhibit limited ability to stop energetic projectile nuclei and to dissipate the imparted linear momentum. Substantial matter overlap (“neck”) between projectile and target nuclei, which is observed at elevated collision energies, can be stretched considerably and break at several places simultaneously. These results provide new testing grounds for microscopic theory of the cohesion of nuclear matter. This property has remained elusive, even though the elementary nucleon-nucleon forces are well known since some time. Technical R&D has resulted in a detailed characterization of a novel plastic material, which can now be used in the design of sensitive diagnostic systems for various types of radio-activity. Innovative application of powerful laser systems has produced intense, controllable sources of exotic particle radioactivity for nuclear investigations. Several students have received their Ph.D. degree in experimental nuclear science for their work on basic nuclear research or R&D projects.

  1. GREEN PEA GALAXIES REVEAL SECRETS OF Lyα ESCAPE

    SciTech Connect

    Yang, Huan; Wang, Junxian; Malhotra, Sangeeta; Rhoads, James E.; Gronke, Max; Dijkstra, Mark; Jaskot, Anne; Zheng, Zhenya E-mail: huan.y@asu.edu E-mail: James.Rhoads@asu.edu

    2016-04-01

    We analyze archival Lyα spectra of 12 “Green Pea” galaxies observed with the Hubble Space Telescope, model their Lyα profiles with radiative transfer models, and explore the dependence of the Lyα escape fraction on various properties. Green Pea galaxies are nearby compact starburst galaxies with [O iii] λ5007 equivalent widths (EWs) of hundreds of Å. All 12 Green Pea galaxies in our sample show Lyα lines in emission, with an Lyα EW distribution similar to high-redshift Lyα emitters. Combining the optical and UV spectra of Green Pea galaxies, we estimate their Lyα escape fractions and find correlations between Lyα escape fraction and kinematic features of Lyα profiles. The escape fraction of Lyα in these galaxies ranges from 1.4% to 67%. We also find that the Lyα escape fraction depends strongly on metallicity and moderately on dust extinction. We compare their high-quality Lyα profiles with single H i shell radiative transfer models and find that the Lyα escape fraction anticorrelates with the derived H i column densities. Single-shell models fit most Lyα profiles well, but not the ones with the highest escape fractions of Lyα. Our results suggest that low H i column density and low metallicity are essential for Lyα escape and make a galaxy an Lyα emitter.

  2. Role of the Ascorbate-Glutathione Cycle of Mitochondria and Peroxisomes in the Senescence of Pea Leaves1

    PubMed Central

    Jiménez, Ana; Hernández, José A.; Pastori, Gabriela; del Río, Luis A.; Sevilla, Francisca

    1998-01-01

    We investigated the relationship between H2O2 metabolism and the senescence process using soluble fractions, mitochondria, and peroxisomes from senescent pea (Pisum sativum L.) leaves. After 11 d of senescence the activities of Mn-superoxide dismutase, dehydroascorbate reductase (DHAR), and glutathione reductase (GR) present in the matrix, and ascorbate peroxidase (APX) and monodehydroascorbate reductase (MDHAR) activities localized in the mitochondrial membrane, were all substantially decreased in mitochondria. The mitochondrial ascorbate and dehydroascorbate pools were reduced, whereas the oxidized glutathione levels were maintained. In senescent leaves the H2O2 content in isolated mitochondria and the NADH- and succinate-dependent production of superoxide (O2·−) radicals by submitochondrial particles increased significantly. However, in peroxisomes from senescent leaves both membrane-bound APX and MDHAR activities were reduced. In the matrix the DHAR activity was enhanced and the GR activity remained unchanged. As a result of senescence, the reduced and the oxidized glutathione pools were considerably increased in peroxisomes. A large increase in the glutathione pool and DHAR activity were also found in soluble fractions of senescent pea leaves, together with a decrease in GR, APX, and MDHAR activities. The differential response to senescence of the mitochondrial and peroxisomal ascorbate-glutathione cycle suggests that mitochondria could be affected by oxidative damage earlier than peroxisomes, which may participate in the cellular oxidative mechanism of leaf senescence longer than mitochondria. PMID:9847106

  3. Reciprocal translocations in grass pea (Lathyrus sativus L.): pattern of transmission, detection of multiple interchanges and their independence.

    PubMed

    Talukdar, Dibyendu

    2010-01-01

    Grass pea (Lathyrus sativus L.) is a diploid crop having 2n = 14 chromosomes. Four different plant types, heterozygous for reciprocal translocation (RT), RT-1, RT-2, RT-3, and RT-4 showing pollen semisterility, were isolated in induced mutant population of three varieties of grass pea. Transmission rate of these 4 translocations was studied in advanced selfed and intercrossed progenies, whereas multiple chromosomal interchanges were detected by the pattern of chromosomal ring formation at meiosis I in the F(1) progeny of RTxRT. RT transmitted at an average of 48.89% in selfed progeny and along with normal plants, RT and translocation homozygotes produced some trisomic plants in the segregation progeny. RT-3 showing maximum frequency (68.20%) of zig-zag or alternate chromosome orientation exhibited highest transmission (55.20%). Presence of a ring of 6 or 2 rings of 4 chromosomes in F(1) double heterozygotes obtained from RTxRT suggested involvement of one common chromosome in translocations of RT-1, RT-2, and RT-4 but a different one in common between RT-2 and RT-3. No common chromosome, however, was shared by RT-3 and RT-4. Thus, 5 out of 7 chromosomes are involved in the present RTs, and one of the translocated chromosomes in RT-1 line was always associated with nucleolar organizing region.

  4. Fusarial wilt control and growth promotion of pigeon pea through bioactive metabolites produced by two plant growth promoting rhizobacteria.

    PubMed

    Dutta, S; Morang, P; Nishanth Kumar, S; Dileep Kumar, B S

    2014-03-01

    The bioactive metabolites produced by two plant growth promoting rhizobacteria strains, a Pseudomonas aeruginosa strain RRLJ 04 and a Bacillus cereus strain BS 03, which showed growth promotion and disease control in pigeon pea against Fusarium udum, were isolated and screened for their efficacy to control fusarial wilt of pigeon pea under gnotobiotic and nursery condition. Bioactive metabolites viz., BM 1 and BM 2 from RRLJ 04 and BM 3 from BS 03 also showed in vitro antibiosis against F. udum. Seeds treated with 50 μl seed⁻¹ of BM 1, 30 μl seed⁻¹ of BM 2 and 70 μl seed⁻¹ of BM 3 and grown in pathogen infested soil showed suppression of wilt disease besides growth enhancement. Per cent disease control was 90 % with BM 2 application as compared to 87 and 83 %, respectively in BM 1 and BM 3 after 90 days of growth. BM 2 treated plants were more resistant to the pathogen as compared to the other fractions tested. Mycelial dry weight was found to be reduced on treatment with the bioactive metabolites. Formation of chlamydospore-like structures was observed in the pathogen mycelium treated with BM 3. The analytical studies confirmed that two of these metabolites are phenazine derivatives.

  5. P3N-PIPO, a Frameshift Product from the P3 Gene, Pleiotropically Determines the Virulence of Clover Yellow Vein Virus in both Resistant and Susceptible Peas

    PubMed Central

    Suzuki, Haruka; Miyashita, Yuri; Choi, Sun Hee; Hisa, Yusuke; Rihei, Shunsuke; Shimada, Ryoko; Jeon, Eun Jin; Abe, Junya; Uyeda, Ichiro

    2016-01-01

    ABSTRACT Peas carrying the cyv1 recessive resistance gene are resistant to clover yellow vein virus (ClYVV) isolates No.30 (Cl-No.30) and 90-1 (Cl-90-1) but can be infected by a derivative of Cl-90-1 (Cl-90-1 Br2). The main determinant for the breaking of cyv1 resistance by Cl-90-1 Br2 is P3N-PIPO produced from the P3 gene via transcriptional slippage, and the higher level of P3N-PIPO produced by Cl-90-1 Br2 than by Cl-No.30 contributes to the breaking of resistance. Here we show that P3N-PIPO is also a major virulence determinant in susceptible peas that possess another resistance gene, Cyn1, which does not inhibit systemic infection with ClYVV but causes hypersensitive reaction-like lethal systemic cell death. We previously assumed that the susceptible pea cultivar PI 226564 has a weak allele of Cyn1. Cl-No.30 did not induce cell death, but Cl-90-1 Br2 killed the plants. Our results suggest that P3N-PIPO is recognized by Cyn1 and induces cell death. Unexpectedly, heterologously strongly expressed P3N-PIPO of Cl-No.30 appears to be recognized by Cyn1 in PI 226564. The level of P3N-PIPO accumulation from the P3 gene of Cl-No.30 was significantly lower than that of Cl-90-1 Br2 in a Nicotiana benthamiana transient assay. Therefore, Cyn1-mediated cell death also appears to be determined by the level of P3N-PIPO. The more efficiently a ClYVV isolate broke cyv1 resistance, the more it induced cell death systemically (resulting in a loss of the environment for virus accumulation) in susceptible peas carrying Cyn1, suggesting that antagonistic pleiotropy of P3N-PIPO controls the resistance breaking of ClYVV. IMPORTANCE Control of plant viral disease has relied on the use of resistant cultivars; however, emerging mutant viruses have broken many types of resistance. Recently, we revealed that Cl-90-1 Br2 breaks the recessive resistance conferred by cyv1, mainly by accumulating a higher level of P3N-PIPO than that of the nonbreaking isolate Cl-No.30. Here we show that a

  6. P3N-PIPO, a Frameshift Product from the P3 Gene, Pleiotropically Determines the Virulence of Clover Yellow Vein Virus in both Resistant and Susceptible Peas.

    PubMed

    Atsumi, Go; Suzuki, Haruka; Miyashita, Yuri; Choi, Sun Hee; Hisa, Yusuke; Rihei, Shunsuke; Shimada, Ryoko; Jeon, Eun Jin; Abe, Junya; Nakahara, Kenji S; Uyeda, Ichiro

    2016-08-15

    Peas carrying the cyv1 recessive resistance gene are resistant to clover yellow vein virus (ClYVV) isolates No.30 (Cl-No.30) and 90-1 (Cl-90-1) but can be infected by a derivative of Cl-90-1 (Cl-90-1 Br2). The main determinant for the breaking of cyv1 resistance by Cl-90-1 Br2 is P3N-PIPO produced from the P3 gene via transcriptional slippage, and the higher level of P3N-PIPO produced by Cl-90-1 Br2 than by Cl-No.30 contributes to the breaking of resistance. Here we show that P3N-PIPO is also a major virulence determinant in susceptible peas that possess another resistance gene, Cyn1, which does not inhibit systemic infection with ClYVV but causes hypersensitive reaction-like lethal systemic cell death. We previously assumed that the susceptible pea cultivar PI 226564 has a weak allele of Cyn1 Cl-No.30 did not induce cell death, but Cl-90-1 Br2 killed the plants. Our results suggest that P3N-PIPO is recognized by Cyn1 and induces cell death. Unexpectedly, heterologously strongly expressed P3N-PIPO of Cl-No.30 appears to be recognized by Cyn1 in PI 226564. The level of P3N-PIPO accumulation from the P3 gene of Cl-No.30 was significantly lower than that of Cl-90-1 Br2 in a Nicotiana benthamiana transient assay. Therefore, Cyn1-mediated cell death also appears to be determined by the level of P3N-PIPO. The more efficiently a ClYVV isolate broke cyv1 resistance, the more it induced cell death systemically (resulting in a loss of the environment for virus accumulation) in susceptible peas carrying Cyn1, suggesting that antagonistic pleiotropy of P3N-PIPO controls the resistance breaking of ClYVV. Control of plant viral disease has relied on the use of resistant cultivars; however, emerging mutant viruses have broken many types of resistance. Recently, we revealed that Cl-90-1 Br2 breaks the recessive resistance conferred by cyv1, mainly by accumulating a higher level of P3N-PIPO than that of the nonbreaking isolate Cl-No.30. Here we show that a susceptible pea line

  7. The pharmacokinetics of phenylethyl alcohol (PEA): safety evaluation comparisons in rats, rabbits, and humans.

    PubMed

    Politano, Valerie T; Diener, Robert M; Christian, Mildred S; Hawkins, David R; Ritacco, Gretchen; Api, Anne Marie

    2013-01-01

    The present studies were conducted to compare the dermal absorption, plasma pharmacokinetics, and excretion of phenylethyl alcohol (PEA) by pregnant and nonpregnant rats, rabbits, and humans. The PEA is a natural fragrance material that is widely used in perfumes, soaps, and lotions and is a major ingredient of natural rose oil. Following dermal (430, 700, or 1400 mg/kg body weight [bw]), gavage (430 mg/kg bw), or dietary (430 mg/kg bw) administration of PEA to rats, plasma concentrations of PEA were found to be low regardless of the route of administration. The plasma concentrations of phenylacetic acid (PAA, the major metabolite of PEA) greatly exceeded the concentrations of PEA and were highest after gavage, followed by dermal then dietary administration. Absorption, distribution, metabolism, and excretion were compared following topical application of ¹⁴C-labeled PEA to rats, rabbits, and humans (specific activities of dosing solutions: 58-580, 164, and 50 µCi/mL, respectively). In rabbits, the plasma concentration-time profile for PAA was markedly prolonged compared to rats or humans. In humans, only 7.6% of the applied dose of PEA was absorbed, versus 77% in rats and 50% in rabbits. Based on a human dermal systemic exposure of 0.3 mg/kg per day from the use of multiple consumer personal care products containing PEA, a rat dermal no observed adverse effect level of 70 mg/kg per day, and the percentage of dose absorbed in humans, the margin of safety exceeds 2600 concluding that, under normal fragrance use conditions, PEA is not a developmental toxicity hazard for humans.

  8. The xylose-rich pectins from pea hulls.

    PubMed

    Renard, C M; Weightman, R M; Thibault, J F

    1997-08-01

    The hot acid extract of pea hull, HSP, was rich in galacturonic acid, arabinose and xylose. It was fractionated by copper precipitation followed by ion-exchange chromatography. The copper-soluble fraction represented 26% of HSP and was mostly composed of an arabinan with a low degree of branching, some heteroxylans and a glucan, probably starch. The copper-precipitate (74% of HSP) contained pectins and some residual arabinan, xylan, glucan and mannan. One of the pectic fractions was rich in terminal xylose and fucose; it could be partially degraded by endo-polygalacturonase but not by endo-xylanase and seemed to contain xylogalacturonans.

  9. Lipogenesis and Redox Balance in Nitrogen-Fixing Pea Bacteroids.

    PubMed

    Terpolilli, Jason J; Masakapalli, Shyam K; Karunakaran, Ramakrishnan; Webb, Isabel U C; Green, Rob; Watmough, Nicholas J; Kruger, Nicholas J; Ratcliffe, R George; Poole, Philip S

    2016-10-15

    Within legume root nodules, rhizobia differentiate into bacteroids that oxidize host-derived dicarboxylic acids, which is assumed to occur via the tricarboxylic acid (TCA) cycle to generate NAD(P)H for reduction of N2 Metabolic flux analysis of laboratory-grown Rhizobium leguminosarum showed that the flux from [(13)C]succinate was consistent with respiration of an obligate aerobe growing on a TCA cycle intermediate as the sole carbon source. However, the instability of fragile pea bacteroids prevented their steady-state labeling under N2-fixing conditions. Therefore, comparative metabolomic profiling was used to compare free-living R. leguminosarum with pea bacteroids. While the TCA cycle was shown to be essential for maximal rates of N2 fixation, levels of pyruvate (5.5-fold reduced), acetyl coenzyme A (acetyl-CoA; 50-fold reduced), free coenzyme A (33-fold reduced), and citrate (4.5-fold reduced) were much lower in bacteroids. Instead of completely oxidizing acetyl-CoA, pea bacteroids channel it into both lipid and the lipid-like polymer poly-β-hydroxybutyrate (PHB), the latter via a type III PHB synthase that is active only in bacteroids. Lipogenesis may be a fundamental requirement of the redox poise of electron donation to N2 in all legume nodules. Direct reduction by NAD(P)H of the likely electron donors for nitrogenase, such as ferredoxin, is inconsistent with their redox potentials. Instead, bacteroids must balance the production of NAD(P)H from oxidation of acetyl-CoA in the TCA cycle with its storage in PHB and lipids. Biological nitrogen fixation by symbiotic bacteria (rhizobia) in legume root nodules is an energy-expensive process. Within legume root nodules, rhizobia differentiate into bacteroids that oxidize host-derived dicarboxylic acids, which is assumed to occur via the TCA cycle to generate NAD(P)H for reduction of N2 However, direct reduction of the likely electron donors for nitrogenase, such as ferredoxin, is inconsistent with their redox

  10. Bisphosphorylated PEA-15 Sensitizes Ovarian Cancer Cells to Paclitaxel by Impairing the Microtubule-Destabilizing Effect of SCLIP

    PubMed Central

    Xie, Xuemei; Bartholomeusz, Chandra; Ahmed, Ahmed A.; Kazansky, Anna; Diao, Lixia; Baggerly, Keith A.; Hortobagyi, Gabriel N.; Ueno, Naoto T.

    2013-01-01

    Paclitaxel is a standard chemotherapeutic agent for ovarian cancer. PEA-15 (phosphoprotein enriched in astrocytes-15 kDa) regulates cell proliferation, autophagy, apoptosis, and glucose metabolism and also mediates AKT-dependent chemoresistance in breast cancer. PEA-15's functions are tightly regulated by its phosphorylation status at Ser104 and Ser116. However, the effect of PEA-15 phosphorylation status on chemosensitivity of cancer cells remains unknown. Here, we tested the hypothesis that PEA-15 phosphorylated at both Ser104 and Ser116 (pPEA-15) sensitizes ovarian cancer cells to paclitaxel. We first found that knockdown of PEA-15 in PEA-15-high-expressing HEY and OVTOKO ovarian cancer cells resulted in paclitaxel resistance, whereas re-expression of PEA-15 in these cells led to paclitaxel sensitization. We next found that SKOV3.ip1-DD cells (expressing phosphomimetic PEA-15) were more sensitive to paclitaxel than SKOV3.ip1-AA cells (expressing nonphosphorylatable PEA-15). Compared to SKOV3.ip1-vector and SKOV3.ip1-AA cells, SKOV3.ip1-DD cells displayed reduced cell viability, inhibited anchorage-independent growth, and augmented apoptosis when treated with paclitaxel. Furthermore, HEY and OVTOKO cells displayed enhanced paclitaxel sensitivity when transiently overexpressing phosphomimetic PEA-15 and reduced paclitaxel sensitivity when transiently overexpressing nonphosphorylatable PEA-15. These results indicate that pPEA-15 sensitizes ovarian cancer cells to paclitaxel. cDNA microarray analysis suggested that SCLIP (SCG10-like protein), a microtubule (MT)-destabilizing protein, is involved in pPEA-15-mediated chemosensitization. We found that reduced expression and possibly posttranslational modification of SCLIP following paclitaxel treatment impaired SCLIP's MT-destabilizing effect, thereby promoting induction of mitotic arrest and apoptosis by paclitaxel. Our findings highlight the importance of pPEA-15 as a promising target for improving the efficacy of

  11. Bisphosphorylated PEA-15 sensitizes ovarian cancer cells to paclitaxel by impairing the microtubule-destabilizing effect of SCLIP.

    PubMed

    Xie, Xuemei; Bartholomeusz, Chandra; Ahmed, Ahmed A; Kazansky, Anna; Diao, Lixia; Baggerly, Keith A; Hortobagyi, Gabriel N; Ueno, Naoto T

    2013-06-01

    Paclitaxel is a standard chemotherapeutic agent for ovarian cancer. PEA-15 (phosphoprotein enriched in astrocytes-15 kDa) regulates cell proliferation, autophagy, apoptosis, and glucose metabolism and also mediates AKT-dependent chemoresistance in breast cancer. The functions of PEA-15 are tightly regulated by its phosphorylation status at Ser104 and Ser116. However, the effect of PEA-15 phosphorylation status on chemosensitivity of cancer cells remains unknown. Here, we tested the hypothesis that PEA-15 phosphorylated at both Ser104 and Ser116 (pPEA-15) sensitizes ovarian cancer cells to paclitaxel. We first found that knockdown of PEA-15 in PEA-15-high expressing HEY and OVTOKO ovarian cancer cells resulted in paclitaxel resistance, whereas re-expression of PEA-15 in these cells led to paclitaxel sensitization. We next found that SKOV3.ip1-DD cells (expressing phosphomimetic PEA-15) were more sensitive to paclitaxel than SKOV3.ip1-AA cells (expressing nonphosphorylatable PEA-15). Compared with SKOV3.ip1-vector and SKOV3.ip1-AA cells, SKOV3.ip1-DD cells displayed reduced cell viability, inhibited anchorage-independent growth, and augmented apoptosis when treated with paclitaxel. Furthermore, HEY and OVTOKO cells displayed enhanced paclitaxel sensitivity when transiently overexpressing phosphomimetic PEA-15 and reduced paclitaxel sensitivity when transiently overexpressing nonphosphorylatable PEA-15. These results indicate that pPEA-15 sensitizes ovarian cancer cells to paclitaxel. cDNA microarray analysis suggested that SCLIP (SCG10-like protein), a microtubule-destabilizing protein, is involved in pPEA-15-mediated chemosensitization. We found that reduced expression and possibly posttranslational modification of SCLIP following paclitaxel treatment impaired the microtubule-destabilizing effect of SCLIP, thereby promoting induction of mitotic arrest and apoptosis by paclitaxel. Our findings highlight the importance of pPEA-15 as a promising target for improving

  12. Properties of nuclei probed by laser light

    NASA Astrophysics Data System (ADS)

    Neugart, Rainer

    2017-03-01

    Viewing objects as small as atomic nuclei by visible light sounds quite unrealistic. However, nuclei usually appear as constituents of atoms whose excitations are indeed associated with the absorption and emission of light. Nuclei can thus interact with light via the atomic system as a whole.

  13. Improvement of nutritive value of grass pea (Lathyrus sativus) seed meal in the formulated diets for rohu, Labeo rohita (Hamilton) fingerlings after fermentation with a fish gut bacterium.

    PubMed

    Ramachandran, S; Bairagi, A; Ray, A K

    2005-09-01

    Eight isonitrogenous (35% crude protein approximately) and isocaloric (4.0 kcalg(-1) approximately) diets were formulated incorporating raw and fermented grass pea (Lathyrus sativus) seed meal at 10%, 20%, 30% and 40% levels by weight into a fish meal based diet and fed to rohu, Labeo rohita, fingerlings for 80 days and fish performance was studied. A particular bacterial strain (Bacillus sp.) isolated from the intestine of adult common carp (Cyprinus carpio) reared in the wild having significant amylolytic, cellulolytic, lipolytic and proteolytic activities were used for fermentation of seed meal for 15 days at 37 degrees C. Fermentation of grass pea seed meal was effective in significantly reducing the crude fibre content and anti-nutritional factors, such as tannins, phytic acid and the neurotoxin, beta-ODAP and enhancing the available free amino acids and fatty acids. In terms of growth response, feed conversion ratio and protein efficiency ratio, 30% fermented grass pea seed meal incorporated diet resulted in significantly (P < 0.05) better performance of rohu fingerlings. In general, growth and feed utilization efficiencies of fish fed diets containing fermented seed meal were superior to those fed diets containing raw seed meal. The apparent protein digestibility (APD) values decreased with increasing levels of raw seed meal in the diets. The APD for raw seed meal was lower at all levels of inclusion in comparison to those for the fermented seed meals. The highest deposition of carcass protein was recorded in fish fed the diet containing 40% fermented seed meal. The results indicated that fermented grass pea seed meal can be incorporated in carp diets up to 30% level compared to 10% level of raw seed meal.

  14. Anatomy of a nonhost disease resistance response of pea to Fusarium solani: PR gene elicitation via DNase, chitosan and chromatin alterations

    PubMed Central

    Hadwiger, Lee A.

    2015-01-01

    Of the multiplicity of plant pathogens in nature, only a few are virulent on a given plant species. Conversely, plants develop a rapid “nonhost” resistance response to the majority of the pathogens. The anatomy of the nonhost resistance of pea endocarp tissue against a pathogen of bean, Fusarium solani f.sp. phaseoli (Fsph) and the susceptibility of pea to F. solani f sp. pisi (Fspi) has been described cytologically, biochemically and molecular-biologically. Cytological changes have been followed by electron microscope and stain differentiation under white and UV light. The induction of changes in transcription, protein synthesis, expression of pathogenesis-related (PR) genes, and increases in metabolic pathways culminating in low molecular weight, antifungal compounds are described biochemically. Molecular changes initiated by fungal signals to host organelles, primarily to chromatin within host nuclei, are identified according to source of the signal and the mechanisms utilized in activating defense genes. The functions of some PR genes are defined. A hypothesis based on this data is developed to explain both why fungal growth is suppressed in nonhost resistance and why growth can continue in a susceptible reaction. PMID:26124762

  15. Observation of DNA and protein distributions in mammalian cell nuclei using STXM

    NASA Astrophysics Data System (ADS)

    Ohigashi, Takuji; Ito, Atsushi; Shinohara, Kunio; Tone, Shigenobu; Kado, Masataka; Inagaki, Yuichi; Wang, Yu-Fu; Kosugi, Nobuhiro

    2016-01-01

    A whole A549 cell and isolated nuclei of HeLa S3 cells in the apoptotic process were investigated by using a scanning transmission X-ray microscope (STXM) in the UVSOR Synchrotron (Okazaki, Japan). Near edge X-ray absorption fine structures (NEXAFS) of DNA and histone in the N K-edge region were measured as reference and their distribution in the nuclei was determined by using these reference spectra. The four stages of the apoptosis were successfully distinguished.

  16. Physical processing of cometary nuclei

    NASA Technical Reports Server (NTRS)

    Weissman, Paul R.; Stern, S. Alan

    1989-01-01

    Cometary nuclei were formed far from the Sun in the colder regions of the solar nebula, and have been stored in distant orbits in the Oort cloud over most of the history of the solar system. It had been thought that this benign environment would preserve comets in close to their original pristine state. However, recent studies have identified a number of physical processes that have likely acted to modify cometary nuclei in a variety of significant ways. It is important to consider all of these possible processes, both in deciding on a site on the nucleus for collection of cometary samples, and in interpreting the results of analyses of returned cometary samples. Although it can no longer be said that comets are pristine samples of original solar nebula material, they are still the best obtainable samples of that unique period in the formation of the planetary system.

  17. Radio characteristics of galactic nuclei

    NASA Astrophysics Data System (ADS)

    Condon, J. J.

    1986-02-01

    Radio characteristics of galactic nuclei, providing such unique information as spectral data on source variability, and the long-term history of the central engine and its duration of activity and total energy, are reviewed. The compact radio source characteristics are complicated by orientation-dependent relativistic beaming and by refractive focusing in the interstellar medium. Incoherent synchrotron radiation is thought to be the emission mechanism, with the result that synchrotron self-absorption in compact sources hides the central engine from direct radio observation. However, the history revealed by the extended jets and lobes of radio galaxies and quasars favors a single massive object not supported by radiation pressure, either a spinar or a black hole, as the energy source in radio-galaxy nuclei.

  18. Environmental properties related to active galactic nuclei

    NASA Astrophysics Data System (ADS)

    Manzer, Lianne H.

    There continues to be significant controversy regarding the mechanisms responsible for the initiation of activity in galactic nuclei. It is well understood that the non-thermal energy produced by an AGN is due to accretion onto a supermassive black hole. It has not yet been determined, however, what leads particular galaxies to become active. An accurate exploration into what triggers an AGN demands an analysis of a large sample of galaxies across a diverse set of environments. In this work, we investigate possible environmental influences by carrying out a statistical investigation of galaxy groups. Using the catalogue of Yang et al. (2007), in which groups of galaxies containing between 2 and 20 members with redshifts between 0.01 -- 0.20 were taken from the Sloan Digital Sky Survey, we investigate the fraction of active galactic nuclei (AGN) within these groups and compare it to the sample of isolated galaxies also obtained from Yang et al. (2007). After correcting our spectroscopic data for extinction and underlying stellar absorption, we classify the galaxy sample using relevant emission-line ratios. We propose an alternate method for classifying emission-line galaxies, including AGN, which builds upon standard diagnostic utilities used for optical classification and includes uncertainties. Such classification probabilities offer a more robust and consistent method of investigating the effect of group environments with galaxy type. We find our sample to be a fair representation of the local universe by comparing the luminosity function of our entire data set to that of Blanton et al. (2001), Blanton et al. (2003b), and Montero-Dorta & Prada (2009). The evidence also suggests that the luminosity function of galaxies differs between isolated galaxies and galaxies in groups. We find a significant increase in the fraction of AGNs identified in grouped environments. On the other hand, we find a higher fraction of starforming galaxies within isolated systems. We

  19. Geometric symmetries in light nuclei

    NASA Astrophysics Data System (ADS)

    Bijker, R.

    2017-06-01

    The algebraic cluster model is is applied to study cluster states in the nuclei12C and16O. The observed level sequences can be understood in terms of the underlying discrete symmetry that characterizes the geometrical configuration of the α-particles, i.e. an equilateral triangle for12C, and a regular tetrahedron for16O. The structure of rotational bands provides a fingerprint of the underlying geometrical configuration of α-particles.

  20. Direct Reactions with Exotic Nuclei

    SciTech Connect

    Baur, G.; Typel, S.

    2005-10-14

    We discuss recent work on Coulomb dissociation and an effective-range theory of low-lying electromagnetic strength of halo nuclei. We propose to study Coulomb dissociation of a halo nucleus bound by a zero-range potential as a homework problem. We study the transition from stripping to bound and unbound states and point out in this context that the Trojan-Horse method is a suitable tool to investigate subthreshold resonances.