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PubMed1974

Control of Diurnal Variations in Photosynthetic Products: II. Nitrate Reductase Activity.

Nitrate accumulates in the leaves of Capsicum annuum L. cv. California Wonder and the leaf content is dependent on the nitrate level supplied to the roots. There is no consistent diurnal periodicity in the leaf nitrate levels.Nitrate reductase activity exhibits three distinct peaks in the leaves. One in the dark period, a second at the time of the prevailing dawn, and a third 6 hours after the beginning of the photoperiod. The third peak of activity can be induced by a short period of illumination (20 minutes) which is also sufficient to induce the labeling of the amino acid fraction by (14)CO(2).

Journal Article↗
PubMed1977

Plant Cuticles Are Polyelectrolytes with Isoelectric Points around Three.

The isoelectric points of isolated cuticles from Citrus aurantium L. (3.15), Prunus armeniaca L. (3.45), and Pyrus communis L. (2.90) leaves were determined from membrane potentials. At pH values below the isoelectric point, cuticular membranes carry a net positive charge and are permselective to anions (determined using (82)Br(-)). Above the isoelectric point, they carry a net negative charge and are permselective to cations (determined using (24)Na(+)). There are no gradients of fixed charges across the cuticular membranes as indicated by the absence of asymmetry potentials. Positive charges in the membranes originate from residues of basic amino acids of proteins or polypeptides contained in a nonextractable form within the cuticle. The exchange capacity of basic fixed groups in the cuticles of six species (Lycopersicon esculentum Mill., Capsicum annuum L. fruit cuticles, and Brassaia spec. leaf cuticles in addition to the above species) varied between 0.010 and 0.025 meq g(-1) cuticle. Fixed acidic groups were donated by residues of acidic amino acids, polygalacturonic acid, and nonesterified -COOH groups of the cutin polymer. At pH 8, total cation exchange capacity as determined using (45)Ca(2+) varied between 0.26 (Citrus) and 0.30 (apricot) meq g(-1).

Journal Article↗
PubMed1977

Envelope Permeability to Possible Precursors of Carotenoid Biosynthesis during Chloroplast-Chromoplast Transformation.

During the transformation of chloroplasts to chromoplasts in Capsicum annuum L., the permeability of the envelope membranes to possible precursors of carotenoid biosynthesis (acetate, mevalonate, citrate) was tested. The plastids were isolated by gel filtration, and the uptake of labeled compounds was measured by a filtering centrifugation technique, using silicone oil. The different ripening stages were characterized by the ratio of chlorophyll to carotenoid content. The chloroplast membranes were shown to be impermeable to all of the metabolites tested. During the transformation there was a sharp increase of membrane permeability. In the mature chromoplast, the permeability to mevalonate and acetate again decreased to about 20% of the maximum value and reached zero for citrate. The results give evidence that during the transformation of chloroplasts to chromoplasts, precursors for carotenoid biosynthesis are translocated from extraplastidic sites into the plastids, there being possibly incorporated into carotenoids.

Journal Article↗
PubMed1978

Pressure probe technique for measuring water relations of cells in higher plants.

A new method is described for continuously measuring cell turgor pressure (P), hydraulic conductivity (L(p)), and volumetric elastic modulus (epsilon) in higher plant cells, using a pressure probe. This technique permits volume changes, DeltaV, and turgor pressure changes, DeltaP, to be determined with an accuracy of 10(-5) to 10(-6) mul and 3 to 5.10(-2) bar, respectively.The main principle of the new method is the same as the pressure probe developed by Zimmermann and Steudle in which pressure is transmitted to a pressure transducer by means of an oil-filled capillary introduced into the cell. In order to use the pressure probe for small tissue cells, the effective compressible volume of the apparatus has to be sufficiently small in comparison to the volume of the cell itself. This is achieved by accurately fixing the oil/cell sap boundary in the very tip of the microcapillary by means of an electronic feedback mechanism, so that the effective volume of the apparatus is reduced to about 2 to 10% of the cell volume. In this way also, errors arising from compressibility of the apparatus and temperature fluctuations can be excluded.Measurements on tissues cells of Capsicum annuum fruits yield epsilon values of 2 to 25 bar. Furthermore, epsilon can be shown to be a function of both cell turgor pressure and cell volume; epsilon increases with increasing turgor pressure and is higher in larger cells.

Journal Article↗
PubMed1978

Growth, Phosphate Pools, and Phosphate Mobilization of Salt-stressed Sesame and Pepper.

The growth and phosphate mobilization of control and salt-stressed sesame (Sesamum indicum L.) and pepper (Capsicum annuum L.) plants were examined to ascertain whether or not translocation limits growth of salt-stressed plants. Plants were grown in a complete nutrient solution with and without excess salt. One-half of the control and salt-stressed plants were later transferred to phosphate-free culture solution ("-P" plants). Measurements of growth and phosphate pools in leaves indicated that with or without salinity "-P" plants utilized their phosphate reserves to support growth for a time at rates equaling those of plants supplied with phosphate. The results indicate that mobilization was not limiting for growth of salt-stressed plants.Defoliation experiments were performed at a developmental stage when the import of assimilates by the youngest expanding leaves could be changed by removing certain source or sink leaves. These experiments also indicated that phloem transport was not limiting for leaf growth on salt-stressed plants.

Journal Article↗
PubMed1979

Disappearance of porphobilinogen deaminase activity in leaves before the onset of senescence.

The activity of porphobilinogen deaminase was measured in young and senescent or mature leaves of pepper (Capsicum annuum), and poinsettia (Euphorbia pulcherrima). Whereas high activity was found in the crude extracts of the young leaves, almost no activity was found in the extracts of senescent or mature leaves. The decrease in deaminase activity was not due to the presence of an isolatable inhibitor. By purifying the crude enzyme extracts from leaves of different ages on DEAE-cellulose columns it was shown that the decrease in deaminase activity was due to a real decrease in the amount of enzyme. Fruiting also decreased porphobilinogen deaminase activity. Several kinetic constants of the C. annuum deaminase were determined.

Journal Article↗
PubMed1983

Mode of action of plastic film in extending life of lemon and bell pepper fruits by alleviation of water stress.

The mechanism by which seal-packaging individual fruit in high density polyethylene film delays deterioration was investigated with lemon (Citrus limon [L.] Burm. f. cv Eureka) and bell pepper (Capsicum annuum L. cv Maor) fruits. Seal-packaging effects were due to the water-saturated atmosphere in the sealed enclosure around the fruit. Softening of fruit was highly correlated with declining water potential of fruit. Sealing drastically inhibited softening as well as changes in cell wall pectins. Sealing also delayed disintegration of membrane as shown by the inhibited leakage of amino acids, in particular, and electrolytes in general. All these effects of sealing were prevented or reduced by including hygroscopic CaCl(2) in the sealed enclosure which reduced the ambient humidity. Furthermore, some of these effects of sealing could be achieved also by maintaining nonsealed fruit in water-saturated atmosphere. Sealing effects could not be related to a possible ;modified atmosphere' mechanism in O(2), CO(2), or ethylene. This work supports the hypothesis that the mode of action of sealing in the polyethylene relates to the alleviation of water stress which exists in harvested fruit.

Journal Article↗
PubMed1986

Accumulation of Putrescine during Chilling Injury of Fruits.

Putrescine (Put) increased 68% in lemon (Citrus limon (L.) Burm. f. cv Bearss) flavedo, 39% in grapefruit (C. paradisi Macf. cv Marsh) flavedo, 49% in grapefruit juice, and 149% in pepper (Capsicum annuum L. cv Early Calwonder) pericarp when fruits were stored at chilling temperatures. In lemon flavedo, the coefficient of correlation (r(2)) between Put concentration with severity of chilling was 0.90 and Put levels almost doubled; the injury index going from 1 to 2 units. Pepper pericarp, which was the most chilling-sensitive tissue tested (injury index going from 1 to 3.8 units), showed the greatest difference in Put accumulation (166 to 413 nanomoles per gram fresh weight) between storage temperatures of 7.2 and 1 degrees C. The least difference (338 to 470 nanomoles per gram fresh weight) was found in grapefruit flavedo between storage temperatures of 15.5 and 4.4 degrees C; the injury index going from 1 to 1.3 units.

Journal Article↗
PubMed1986

Species variation in the predawn inhibition of ribulose-1,5-bisphosphate carboxylase/oxygenase.

The activity of ribulose-1,5-bisphosphate carboxylase/oxygenase was measured in extracts of leaves collected before dawn (predawn activity, pa) and at midday (midday activity, ma). Twenty-three of the 37 species examined showed a pa/ma ratio (</=0.75, while only Capsicum frutescens, Cucumis sativa, Glycine max, Nicotiana tabacum, Vigna unguiculata, and 3 Solanum species showed a pa/ma ratio </=0.5. Phaseolus vulgaris consistently showed a pa/ma ratio of </=0.1. Activities and pa/ma ratios of the same species grown in the United States and the United Kingdom were very similar. Gel filtration of extracts before assay had no effect on the observed activities and the pa/ma ratios. These data are consistent with the hypothesis that in a number of species the enzyme is partially inhibited following the night period by the presence of a tight-binding inhibitor.

Journal Article↗
PubMed1987

Effect of Acid treatment of plant cuticles on sorption of selected auxins.

Sorption characteristics of 2-(1-naphthyl)acetic acid (NAA), 2-(1-naphthyl)acetamide (NAAm), and 2,4-dichlorophenoxyacetic acid (2,4-D) were determined for cuticles enzymically isolated from mature tomato (Lycopersicon esculentum Mill. cv Sprinter) and pepper (Capsicum annuum L.) fruit. Sorption equilibrium for NAA and 2,4-D by tomato cuticular membranes (CM) and dewaxed cuticular membranes (DCM) was achieved within 24 hours at 25 degrees C. The average K (partition coefficient) values for NAA in tomato CM and DCM were 166 and 204, respectively, whereas the corresponding K values for 2,4-D were 292 and 383, respectively. Sorption equilibrium for 2,4-D and NAA in pepper cuticles was not achieved after 18 and 63 days, respectively. Sorption equilibrium for NAAm in tomato and pepper CM and DCM was attained within 48 hours. Acid pretreatment (2.0 n HCl, 10 minutes) had no effect on NAA, 2,4-D, or NAAm sorption by tomato CM and DCM, or on NAAm sorption by pepper CM and DCM. Acid pretreatment of pepper CM and DCM led to slightly lower K(pH) (apparent partition coefficient) values for both NAA and 2,4-D. More significantly, sorption equilibrium for NAA and 2,4-D in pepper CM and DCM was achieved within 24 hours after acid treatment.

Journal Article↗
PubMed1987

The Effect of Temperature on the Occurrence of O(2) and CO(2) Insensitive Photosynthesis in Field Grown Plants.

The sensitivity of photosynthesis to O(2) and CO(2) was measured in leaves from field grown plants of six species (Phaseolus vulgaris, Capsicum annuum, Lycopersicon esculentum, Scrophularia desertorum, Cardaria draba, and Populus fremontii) from 5 degrees C to 35 degrees C using gas-exchange techniques. In all species but Phaseolus, photosynthesis was insensitive to O(2) in normal air below a species dependent temperature. CO(2) insensitivity occurred under the same conditions that resulted in O(2) insensitivity. A complete loss of O(2) sensitivity occurred up to 22 degrees C in Lycopersicon but only up to 6 degrees C in Scrophularia. In Lycopersicon and Populus, O(2) and CO(2) insensitivity occurred under conditions regularly encountered during the cooler portions of the day. Because O(2) insensitivity is an indicator of feedback limited photosynthesis, these results indicate that feedback limitations can play a role in determining the diurnal carbon gain in the field. At higher partial pressures of CO(2) the temperature at which O(2) insensitivity occurred was higher, indicating that feedback limitations in the field will become more important as the CO(2) concentration in the atmosphere increases.

Journal Article↗
PubMed1987

Effect of altered sink: source ratio on photosynthetic metabolism of source leaves.

When seven crop species were grown under identical environmental conditions, decreased sink:source ratio led to a decreased photosynthetic rate within 1 to 3 days in Cucumis sativus L., Gossypium hirsutum L., and Raphanus sativus L., but not in Capsicum annuum L., Solanum melongena L., Phaseolus vulgaris L., or Ricinus communis L. The decrease was not associated with stomatal closure. In cotton and cucumber, sink removal led to an increase in starch and sugar content, in glucose 6-phosphate and fructose 6-phosphate pools, and in the proportion of (14)C detected in sugar phosphates and UDPglucose following (14)CO(2) supply. When mannose was supplied to leaf discs to sequester cytoplasmic inorganic phosphate, promotion of starch synthesis, and inhibition of CO(2) fixation, were observed in control discs, but not in discs from treated plants. Phosphate buffer reduced starch synthesis in the latter, but not the former discs. The findings suggest that sink removal led to a decreased ratio inorganic phosphate:phosphorylated compounds. In beans (14)C in sugar phosphates increased following sink removal, but without sucrose accumulation, suggesting tighter feedback control of sugar level. Starch accumulated to higher levels than in the other plants, but CO(2) fixation rate was constant for several days.

Journal Article↗
PubMed1990

Variation among Species in the Temperature Dependence of the Reappearance of Variable Fluorescence following Illumination.

The relationship between the thermal dependence of the reappearance of chlorophyll variable fluorescence following illumination and temperature dependence of the apparent Michaelis constant (K(m)) of NADH hydroxypyruvate reductase for NADH was investigated in cool and warm season plant species. Brancker SF-20 and SF-30 fluorometers were used to evaluate induced fluorescence transients from detached leaves of wheat (Triticum aestivum L. cv TAM-101), cotton (Gossypium hirsutum L. cv Paymaster 145), tomato (Lycopersicon esculentum cv Del Oro), bell pepper (Capsicum annuum L. cv California Wonder), and petunia (Petunia hybrida cv. Red Sail). Following an illumination period at 25 degrees C, the reappearance of variable fluorescence during a dark incubation was determined at 5 degrees C intervals from 15 degrees C to 45 degrees C. Variable fluorescence recovery was normally distributed with the maximum recovery observed at 20 degrees C in wheat, 30 degrees C in cotton, 20 degrees C to 25 degrees C in tomato, 30 to 35 degrees C in bell pepper and 25 degrees C in petunia. Comparison of the thermal response of fluorescence recovery with the temperature sensitivity of the apparent K(m) of hydroxypyruvate reductase for NADH showed that the range of temperatures providing fluorescence recovery corresponded with those temperatures providing the minimum apparent K(m) values (viz. the thermal kinetic window).

Journal Article↗
PubMed1992

Posttranslational modifications in the amino- terminal region of the large subunit of ribulose- 1,5-bisphosphate carboxylase/oxygenase from several plant species.

A combination of limited tryptic proteolysis, reverse phasehigh performance liquid chromatography, Edman degradative sequencing, amino acid analysis, and fast-atom bombardment mass-spectrometry was used to remove and identify the first 14 to 18 N-terminal amino acid residues of the large subunit of higher plant-type ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) from Chlamydomonas reinhardtii, Marchantia polymorpha, pea (Pisum sativum), tomato (Lycopersicon esculentum), potato (Solanum tuberosum), pepper (Capsicum annuum), soybean (Glycine max), petunia (Petunia x hybrida), cowpea (Vigna sinensis), and cucumber (Cucumis sativus) plants. The N-terminal tryptic peptide from acetylated Pro-3 to Lys-8 of the large subunit of Rubisco was identical in all species, but the amino acid sequence of the penultimate N-terminal tryptic peptide varied. Eight of the 10 species examined contained a trimethyllysyl residue at position 14 in the large subunit of Rubisco, whereas Chlamydomonas and Marchantia contained an unmodified lysyl residue at this position.

Journal Article↗
PubMed2005

Molecular genetic evidence for the role of SGT1 in the intramolecular complementation of Bs2 protein activity in Nicotiana benthamiana.

Pepper plants (Capsicum annuum) containing the Bs2 resistance gene are resistant to strains of Xanthomonas campestris pv vesicatoria (Xcv) expressing the bacterial effector protein AvrBs2. AvrBs2 is delivered directly to the plant cell via the type III protein secretion system (TTSS) of Xcv. Upon recognition of AvrBs2 by plants expressing the Bs2 gene, a signal transduction cascade is activated leading to a bacterial disease resistance response. Here, we describe a novel pathosystem that consists of epitope-tagged Bs2-expressing transgenic Nicotiana benthamiana plants and engineered strains of Pseudomonas syringae pv tabaci that deliver the effector domain of the Xcv AvrBs2 protein via the TTSS of P. syringae. This pathosystem has allowed us to exploit N. benthamiana as a model host plant to use Agrobacterium tumefaciens-mediated transient protein expression in conjunction with virus-induced gene silencing to validate genes and to identify protein interactions required for the expression of plant host resistance. In this study, we demonstrate that two genes, NbSGT1 and NbNPK1, are required for the Bs2/AvrBs2-mediated resistance responses but that NbRAR1 is not. Protein localization studies in these plants indicate that full-length Bs2 is primarily localized in the plant cytoplasm. Three protein domains of Bs2 have been identified: the N terminus, a central nucleotide binding site, and a C-terminal Leu-rich repeat (LRR). Co-immunoprecipitation studies demonstrate that separate epitope-tagged Bs2 domain constructs interact in trans specifically in the plant cell. Co-immunoprecipitation studies also demonstrate that an NbSGT1-dependent intramolecular interaction is required for Bs2 function. Additionally, Bs2 has been shown to associate with SGT1 via the LRR domain of Bs2. These data suggest a role for SGT1 in the proper folding of Bs2 or the formation of a Bs2-SGT1-containing protein complex that is required for the expression of bacterial disease resistance.

Arabidopsis Proteins↗
PubMed2005

The conserved Xanthomonas campestris pv. vesicatoria effector protein XopX is a virulence factor and suppresses host defense in Nicotiana benthamiana.

Nicotiana benthamiana leaves display a visible plant cell death response when infiltrated with a high titer inoculum of the non-host pathogen, Xanthomonas campestris pv. vesicatoria (Xcv). This visual phenotype was used to identify overlapping cosmid clones from a genomic cosmid library constructed from the Xcv strain, GM98-38. Individual cosmid clones from the Xcv library were conjugated into X. campestris pv. campestris (Xcc) and exconjugants were scored for an altered visual high titer inoculation response in N. benthamiana. The molecular characterization of the cosmid clones revealed that they contained a novel gene, xopX, that encodes a 74-kDa type III secretion system (TTSS) effector protein. Agrobacterium-mediated transient expression of XopX in N. benthamiana did not elicit the plant cell death response although detectable XopX protein was produced. Interestingly, the plant cell death response occurred when the xopX Agrobacterium-mediated transient expression construct was co-inoculated with strains of either XcvDeltaxopX or Xcc, both lacking xopX. The co-inoculation complementation of the plant cell death response also depends on whether the Xanthomonas strains contain an active TTSS. Transgenic 35S-xopX-expressing N. benthamiana plants also have the visible plant cell death response when inoculated with the non-xopX-expressing strains XcvDeltaxopX and Xcc. Unexpectedly, transgenic 35S-xopX N. benthamiana plants displayed enhanced susceptibility to bacterial growth of Xcc as well as other non-xopX-expressing Xanthomonas and Pseudomonas strains. This result is also consistent with the increase in bacterial growth on wild type N. benthamiana plants observed for Xcc when XopX is expressed in trans. Furthermore, XopX contributes to the virulence of Xcv on host pepper (Capsicum annuum) and tomato (Lycopersicum esculentum) plants. We propose that the XopX bacterial effector protein targets basic innate immunity in plants, resulting in enhanced plant disease susceptibility.

Amino Acid Sequence↗
PubMed1996

Occupational asthma due to different spices.

We describe a 27-year-old subject who developed rhinitis and asthma symptoms 1 year after starting to prepare a certain kind of sausage. He was previously diagnosed as having allergy to coconut, banana, and kiwi and allergic rhinitis to horse, cat, dog, and cow. A positive immediate skin prick test (SPT) for paprika (dry powder of Capsicum annuum [Solanaceae]), coriander (Coriandrum sativum [umbelliferous]), and mace (shell of nutmeg, Myristica fragrans [Myristicaceae]) at a concentration of 10% (w/v) was obtained. SPT with other sausage ingredients, mites, pollens, and molds were negative. By ELISA, specific IgE antibodies to paprika, coriander, and mace were demonstrated. By ELISA-inhibition assays, a partial cross-reactivity was found among IgE-binding components from paprika and mace. The immunoblot analysis showed two IgE-reactive protein bands able to bind to IgE from mace of 20 and 40 kDa and two other bands from coriander extract of 50 and 56 kDa. No bands were detected from paprika extract. Specific bronchial inhalation challenges showed an immediate asthmatic reaction to extracts from paprika, coriander, and mace with a maximum fall in FEV1 of 26%, 40%, and 31%, respectively, with no late asthmatic reactions. In summary, we demonstrate that inhalation of dust from paprika, coriander, and mace can result in an IgE-mediated reaction to these spices. In this patient, occupational asthma was due to spices from botanically unrelated species.

Adult↗
PubMed1990

Structural investigations and biological activity of inositol sphingophospholipids from Phytophthora capsici.

Inositol sphingophospholipids that protect pepper (Capsicum annuum c.v. Yolo Wonder) against pathogen have been isolated by chromatographic methods from the mycelium of Phytophthora capsici. The structure of the major compound was determined by chemical methods and mass spectrometry. Phosphodiester bond cleavage of the phospholipid by mild alkaline hydrolysis liberated a ceramide which contained a C16-sphingosine. This long-chain base was identified by gas chromatography and mass spectrometry of its trimethylsilyl derivative. One of the amide-linked fatty acids was found to be 4-hydroxy-2 docosenoic acid. Fast-atom-bombardment mass spectrometry and fast-atom-bombardment collison-induced tandem mass spectrometry were used to characterize the ceramide as N(4-hydroxy-2-docosenoyl)C16-sphingosine. These sphingolipids have a protective effect on cotyledons of young peppers against necrotic lesions induced by the pathogen P. capsici.

Ceramides↗