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  • 1
    Electronic Resource
    Electronic Resource
    Partial deglycosylation of an anionic isoperoxidase from peach seeds – effect on enzyme activity, stability and antigenicity (1991)
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 83 (1991), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: An anionic isoperoxidase (EC 1.11.1.7) purified from peach seeds (Prunus persica L. Batsch cv. Merry) was partially deglycosylated by glycopeptidase F (EC 3.2.2.18) treatment. A 40% deglycosylation resulted in an activity loss of 50% when assayed with o-dianisidine. 60% with guaiacol and 78% with 2,2′-azino-bis(3-ethyl)benzethiozoline-6-sulfonic acid (ABTS) as substrate. The indole-3-acetic acid oxidase activity loss was close to 55%. The partially deglycosylated isoperoxidase also showed a higher Km value for H2O2 and higher values for Arrhenius activation energy and enthalpy of activation. There was a decrease in enzyme stability at 4°C after deglycosylation. Native and partially deglycosylated isoperoxidase reacted equally well in an enzyme-linked immunosorbent assay (ELISA) with rabbit polyclonal antibodies raised against the native enzyme. The carbohydrate moiety of this peach seed isoperoxidase appears to be important for enzyme activity and stability.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.1991.tb01293.x
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  • 2
    Electronic Resource
    Electronic Resource
    Structural and physiological changes in the roots of tomato plants over-expressing a basic peroxidase (2003)
    Oxford, UK : Munksgaard International Publishers
    Physiologia plantarum 118 (2003), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Previous studies on the tomato (Lycopersicon esculentum Mill.) peroxidase TPX1, including the development of transgenic tomato over-expressing this gene, supported an involvement of this peroxidase in the synthesis of lignin and suberin. The transgenic plants showed a wilty phenotype at flowering, but the relationship between this role in ligno-suberization and this phenotype was not clear. In the present study a histological approach and the measurement of water-related parameters have been performed in order to obtain an insight into the origin of this phenotype. Clear differences between transgenic and non-transgenic roots were observed in the cross-sections of the basal root zones where secondary growth was evident. The diameter of the xylem vessel was diminished in the transgenic plants. Total area corresponding to xylem in the basal cross-sections decreased 3.9 fold in the transgenic roots. In addition, the radial and outer tangential walls of the exodermis cells were more ligno-suberized in transgenic than in non-transgenic plants. After fruit set, predawn and midday water potentials were lower in transgenic than in-non-transgenic plants. At midday, the stomatal conductance was also lower in the transgenic plants, 494±69 versus 594±60 mmol m−2 s−1. Root hydraulic conductances of the transgenic and non-transgenic plants were 1.4±0.38 and 3.47±0.19 g water min−1 MPa−1, respectively. The results obtained support that the phenotype is caused by the anatomical differences found in the transgenic roots. These differences would be the cause of a increased resistance to water flow in the roots that would negatively affect the water supply to the shoot and, as a consequence, resulted in a decreased water potential in the leaves.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1034/j.1399-3054.2003.00115.x
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  • 3
    Electronic Resource
    Electronic Resource
    Changes in the water binding characteristics of the cell walls from transgenic Nicotiana tabacum leaves with enhanced levels of peroxidase activity (2004)
    Oxford, UK; Malden, USA : Munksgaard International Publishers
    Physiologia plantarum 122 (2004), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Pore size in the cell wall matrix may affect cell wall–water relations, particularly under osmotic stress. Cross linkage of plant cell wall matrix polymers is an important step in the formation of this structure and peroxidases have been proposed to catalyse the cross-linking of phenolic constituents. Transgenic tobacco (Nicotiana tabacum) plants expressing a basic tomato peroxidase gene (TPX2) showed increased apoplastic ferulic acid peroxidase activity in mature leaves. This enhanced activity was not associated with a decreased leaf growth. Differential scanning calorimetry (DSC) of control dried cell walls showed a putative glass transition, after Ca2+ removal, that was absent in the transgenic line. This would indicate that transgenic walls were more rigid. DSC analysis of water-hydrated cell wall preparations distinguished two pools of water, freezable and non-freezable water. The amount of non-freezable water, which corresponds to strongly bound water, was higher in the transgenic line (64 versus 55%). DSC thermograms of the transgenic cell wall were displaced to lower temperatures, and this may be interpreted as the result of a stronger interaction between this freezable water and this wall. Water sorption and desorption isotherms, obtained at relative humidity ranging from 5 to 93%, demonstrated the presence of very strongly bound water in the transgenic cell walls that was absent in controls. Water sorption–desorption hysteresis of the isotherms was evident in the control wall but not in the transgenic line. These changes in cell wall–water interaction seem to be relevant at the organ level because leaf discs of transgenic plants maintained higher relative water content than control discs, at water potentials between −1.05 and−2.31 MPa.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.2004.00429.x
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  • 4
    Electronic Resource
    Electronic Resource
    Biochemical and phenotypical characterization of transgenic tomato plants overexpressing a basic peroxidase (1999)
    Copenhagen : Munksgaard International Publishers
    Physiologia plantarum 106 (1999), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Tomato plants (Lycopersicon esculentum Mill. cv. Pera) were transformed via Agrobacterium tumefaciens with the binary vector pKYLX71 containing a tomato basic peroxidase (EC 1.11.1.7) gene, tpx1, under the control of the cauliflower mosaic virus (CaMV35S) promoter. Transgenic plants showed a 2–5-fold increase in the activity of the peroxidase ionically bound to the cell wall, whereas soluble peroxidase activity remained similar or even lower than wild-type plants. Isoelectric focusing showed the presence of a new isoperoxidase of pI ca 9 in the ionically bound extract. Western blot also showed the presence of a new band at 41 kDa that was absent in the wild-type extract. A 40–220% increment of lignin content of the leaf was found in transgenic plants. Shoot phenotype of transgenic plants was similar to wild type, although under stress, the plants appeared wilted and the new leaves had a reduced area and were thicker than wild-type or older transgenic leaves. The root system was underdeveloped in transgenic plants, but the rooting ability of the stem was not affected by the overexpression of peroxidase. Finally, the morphogenetic response of cotyledon and hypocotyl explants from transgenic plants was evaluated. In the case of cotyledons, the percentage of explants with shoot was not different from wild-type plants. For hypocotyl, one of the transgenic lines showed a 30% reduction in the percentage of shoot organogenesis. The results are discussed in relation to the role of tpx1 in lignin synthesis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1034/j.1399-3054.1999.106401.x
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  • 5
    Electronic Resource
    Electronic Resource
    Purification of an anionic isoperoxidase from peach seeds and its immunological comparison with other anionic isoperoxidases (1990)
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 79 (1990), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A soluble anionic isoperoxidase (EC 1,11,1,7) was purified from peach (Prunus persica L. Batsch cv. Merry) seeds. Purification was achieved by DEAE-Sephacel, Sephacryl S-300 and CM-cellulose chromatography. The purified isoperoxidase de-carboxylated indole-3-acetic acid (S0.5 0.13 mM, Hill coefficient 1.7). Molecular mass, determined by gel filtration and sodium dodecyl sulfate polyacrylamide gel electrophoresis, was ca 60 kDa. Polyclonal antibodies were raised in rabbit against this isoperoxidase. Using immunoprecipitation this isoenzyme was found to be immunologically different from other soluble anionic isoperoxidases isolated from peach seeds.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.1990.tb00035.x
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  • 6
    Electronic Resource
    Electronic Resource
    Peroxidase and IAA oxidase activities and peroxidase isoenzymes in the pericarp of seeded and seedless “Redhaven” peach fruit (1992)
    Springer
    Journal of plant growth regulation 11 (1992), S. 1-6 
    Details
    ISSN: 1435-8107
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Parthenocarpic peach fruit (Prunus persica L. Batsch., cv. Redhaven) were induced with 1-(3-chlorophthalimide)-cyclohexane carboxamide (AC 94377). The activities of soluble, and ionically and covalently bound peroxidase and indole-3-acetic acid (IAA) oxidase in the pericarp of both seeded and parthenocarpic fruit were determined from 21–43 days after anthesis. Seedless fruit grew faster during early stage I and ceased growth earlier than seeded fruit. Total peroxidase and IAA oxidase activities increased with development on both types of fruit, but higher values were found in seedless fruit. The ionic fraction showed the greatest increase for both enzyme activities. Isoperoxidase profile showed new cationic isoenzymes and higher levels of the less anionic isoenzymes in the pericarp of seedless fruit, whereas the seeded fruit contained higher levels of the more acidic isoperoxidases.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00193836
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  • 7
    Electronic Resource
    Electronic Resource
    Shoot regeneration and Agrobacterium-mediated transformation of Fragaria vesca L. (1996)
    Springer
    Plant cell reports 15 (1996), S. 642-646 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary An efficient and reliable method for shoot regeneration from leaf disks of Fragaria vesca L. has been developed. This protocol has been successfully employed to obtain transformed plants using Agrobacterium tumefaciens as gene vector. Murashige and Skoog basal medium supplemented with benzyladenine (4 mg/l) and indole-3-butyric acid (0.25 mg/l) induced the maximum percentage of shoot regeneration (98%) and the highest number of shoot colonies per explant (4.6) after 8 weeks of culture. Isolated shoots would elongate and proliferate when the benzyladenine concentration was lowered to 0.5 mg/l. The established protocol for shoot regeneration was employed to transform leaf disks using Agrobacterium tumefaciens carrying the plasmid pBI121. A 7.7% of the inoculated explants showed kanamycin resistance after 10 weeks of selection in a medium containing 25 mg/l of this antibiotic. The transgenic shoots obtained were rooted in the presence of 25 mg/ kanamycin and successfully acclimatized. The final percentage of transformation obtained based on beta-glucuronidase expression was 6.9%.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00232469
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  • 8
    Electronic Resource
    Electronic Resource
    Shoot regeneration and Agrobacterium-mediated transformation of Fragaria vesca L. (1996)
    Springer
    Plant cell reports 15 (1996), S. 642-646 
    Details
    ISSN: 1432-203X
    Keywords: Abbreviations: BA: benzyladenine; IBA: indole-3-butyric acid; MS: Murashige and Skoog basal medium; LSD: least significant difference; NOS: nopaline synthase promoter, NPTII: neomycin phosphotransferase (EC 2.7.2.95); CaMV35S: cauliflower mosaic virus promoter; GUS: beta-glucuronidase (EC 3.2.1.31); LB: Luria Broth base; CTAB: hexadecil trimethyl ammonium bromide; PCR: polymerase chain reaction; X-gluc: 5-bromo-4-chloro-3-indolyl-glucuronide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary. An efficient and reliable method for shoot regeneration from leaf disks of Fragaria vesca L. has been developed. This protocol has been successfully employed to obtain transformed plants using Agrobacterium tumefaciens as gene vector. Murashige and Skoog basal medium supplemented with benzyladenine (4 mg/l) and indole-3-butyric acid (0.25 mg/l) induced the maximum percentage of shoot regeneration (98%) and the highest number of shoot colonies per explant (4.6) after 8 weeks of culture. Isolated shoots would elongate and proliferate when the benzyladenine concentration was lowered to 0.5 mg/l. The established protocol for shoot regeneration was employed to transform leaf disks using Agrobacterium tumefaciens carrying the plasmid pBI121. A 7.7% of the inoculated explants showed kanamycin resistance after 10 weeks of selection in a medium containing 25 mg/l of this antibiotic. The transgenic shoots obtained were rooted in the presence of 25 mg/l kanamycin and successfully acclimatized. The final percentag of transformation obtained based on beta-glucuronidase expression was 6.9%.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002990050090
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  • 9
    Electronic Resource
    Electronic Resource
    Pollen sporopollenin: degradation and structural elucidation (1999)
    Springer
    Sexual plant reproduction 12 (1999), S. 171-178 
    Details
    ISSN: 1432-2145
    Keywords: Key words Pollen ; Exine ; Sporopollenin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  We report the isolation of purified sporopollenin from pollen grains of different species and its complete solubilization. Exine from Pinus pinaster, Betula alba, Ambrosia elatior and Capsicum annuum was extracted by treatment with hydrogen fluoride in pyridine. These exines were purified from their aromatic moieties and from fatty acids linked by ester bonds using acidolysis and saponification treatments. The biopolymer obtained retains almost completely the shape of the original pollen grain. Fourier-transform infrared spectroscopy analysis of the isolated sporopollenin showed the absence of polysaccharide and phenolic material and the presence of carboxylic acid groups joined to unsaturations and ether linkages. Sporopollenin samples were successfully degraded by exhaustive 24-h ozonolysis at room temperature. Gentle ozonolysis (3 h at 0°C) did not completely degrade the biopolymer. The compounds obtained after exhaustive ozonolysis were analysed by gas chromatography-mass spectrometry. Dicarboxylic acids with a low number of carbon atoms were identified as major components of sporopollenin from P. pinaster, A. elatior and C. annuum, representing 28.8%, 63.2% and 88.5%, respectively, of the total compounds obtained. Fatty acids and n-alkanes also were identified in P. pinaster, A. elatior and B. alba sporopollenin. From the data obtained, an hypothesis about the chemical nature and structural arrangement of the sporopollenin is proposed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004970050189
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  • 10
    Electronic Resource
    Electronic Resource
    Changes in indole-3-acetic acid, indole-3-acetic acid oxidase, and peroxidase isoenzymes in the seeds of developing peach fruits (1989)
    Springer
    Journal of plant growth regulation 8 (1989), S. 255-261 
    Details
    ISSN: 1435-8107
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Changes in indole-3-acetic acid (IAA) content of peach (Prunus persica L. Batsch cv. Merry) seeds were followed during fruit development. The highest concentration of IAA, 2.7 μg/g fresh weight, was found at the beginning of Stage III of fruit development, approximately 50–60 days after anthesis. The IAA-decarboxylating capacity of crude extracts of seeds was also greatest at 55–60 days after anthesis. Four soluble peroxidase isoenzymes were found on anionic electrophoresis. There were no marked changes in two isoenzymes (R f 0.23 and 0.51), which were present in all three stages of fruit growth. There was a marked increase in a band atR f 0.59 between Stages II and III, and a decrease in a band atR f 0.68 from Stages II to III. Neither band (R f 0.59 and 0.68) was present at Stage I.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02021818
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