942 resultados para BARLEY PEROXIDASE
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"Supersedes Technical bulletin no. 907, Classification of barley varieties grown in the United States and Canada in 1945."
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Caption title.
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A full-length cDNA sequence coding for Echinococcus granulosus thioredoxin peroxidase (EgTPx) was isolated from a sheep strain protoscolex cDNA library by immunoscreening using a pool of sera from mice infected with oncospheres. EgTPx expressed as a fusion protein with glutathione S-transferase (GST) exhibited significant thiol-dependent peroxidase activity that protected plasmid DNA from damage by metal-catalyzed oxidation (MCO) in vitro. Furthermore, the suggested antioxidant role for EgTPx was reinforced in an in vivo assay, whereby its expression in BL21 bacterial cells markedly increased the tolerance and survival of the cells to high concentrations of H2O2 compared with controls. Immunolocalization studies revealed that EgTPx was specifically expressed in all tissues of the protoscolex and brood capsules. Higher intensity of labelling was detected in many, but not all, calcareous corpuscle cells in protoscoleces. The purified recombinant EgTPx protein was used to screen sera from heavily infected mice and patients with confirmed hydatid infection. Only a portion of the sera reacted positively with the EgTPx-GST fusion protein in Western blots, suggesting that EgTPx may form antibody-antigen complexes or that responses to the EgTPx antigen may be immunologically regulated. Recombinant EgTPx may prove useful for the screening of specific inhibitors that could serve as new drugs for treatment of hydatid disease. Moreover, given that TPx from different parasitic phyla were phylogenetically distant from host TPx molecules, the development of antiparasite TPx inhibitors that do not react with host TPx might be feasible. (C) 2003 Elsevier B.V. All rights reserved.
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Lines of transgenic tobacco have been generated that are transformed with either the wild-type peanut peroxidase prxPNC2 cDNA, driven by the CaMV3 5S promoter (designated 35S::prxPNC2-WT) or a mutated PNC2 cDNA in which the asparagine residue (Asn(189)) associated with the point of glycan attachment (Asn(189)) has been replaced with alanine (designated 35S::prxPNC2-M). PCR, using genomic DNA as template, has confirmed the integration of the 35S::prxPNC2-WT and 35::prxPNC2-M constructs into the tobacco genome, and western analysis using anti-PNC2 antibodies has revealed that the prxPNC2-WT protein product (PNC2-WT) accumulates with a molecular mass of 34,670 Da, while the prxPNC2-M protein product (PNC2-M) accumulates with a molecular mass of 32,600 Da. Activity assays have shown that both PNC2-WT and PNC2-M proteins accumulate preferentially in the ionically-bound cell wall fraction, with a significantly higher relative accumulation of the PNC2-WT isoenzyme in the ionically-bound fraction when compared with the PNC2-M isoform. Kinetic analysis of the partially purified PNC2-WT isozyme revealed an affinity constant (apparent K-m) of 11.2 mM for the reductor substrate guaiacol and 1.29 mM for H2O2, while values of 11.9 mM and 1.12 mM were determined for the PNC2-M isozyme. A higher Arrenhius activation energy (E,,) was determined for the PNC2-M isozyme (22.9 kJ mol(-1)), when compared with the PNC2-WT isozyme (17.6 kJ mol(-1)), and enzyme assays have determined that the absence of the glycan influences the thermostability of the PNC2-M isozyme. These results are discussed with respect to the proposed roles of N-linked glycans attached to plant peroxidases. (c) 2005 Elsevier Ltd. All rights reserved.
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The free-ion model (FIM) describes iron ( Fe) uptake by barley [ Hordeum vulgare ( L.) 'Grammett'] as being controlled by the activity of the buffered, free, uncomplexed Fe3+ in solution. Chelators' effect on Fe uptake by barley was evaluated and the rate of exchange of Fe between chelators was examined. Barley was grown for two weeks in a low-Fe nutrient solution and transferred to solutions varying in Fe and chelators for 6 h assays. Shoot Fe-59 was higher in barley grown in citrate (7743 and 1928 Fe-59 Bq g(-1)) than in NTA(3220 and 1113 Fe-59 Bq g(-1); P = 0.045) despite similar free-Fe3+ activities. A comparison of Fe uptake by barley from solutions with pFe(3+) activities of 17.1 and 24.6 showed < 5% was from indiscriminate apoplastic-flow uptake (3250 Fe-59 Bq g(-1) vs. 160 Fe-59 Bq g(-1)). Using nutrient solutions from the barley studies but without plants, Fe exchange between chelators and a simulant for the barley phytosiderophore occurred within hours ( for NTA and citrate), or days ( EDTA and HEDTA). Results were similar between the barley and Fe-exchange experiments for the two nutrient-solution treatments where the same Fe3+ activities but different total-Fe concentrations were used: the higher total-Fe treatment resulted in six-fold higher shoot Fe-59, while in the Fe-exchange study that treatment had six-fold more Fe bound to the phytosiderophore simulant after 2 d. Results indicated deviations from the FIM were not explained by indiscriminate-flow uptake, and that sluggish Fe-exchange reactions between chelate and phytosiderophoresimulant, not FIM guidelines, may be more important in explaining Fe uptake from synthetic chelates by Fe-deficient barley.
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Studies on variation, occurrence and distribution of virulence in Pyrenophora teres f. teres are essential to identify effective sources of resistance for net type net blotch. Disease surveys suggested two different stains are prevalent in Western Australia and 13 in all around Australia. Sixty nine barley lines from different breeding groups in Australia and elsewhere were tested against most prevalent pathotypes. Majority of lines have partial to complete resistance while some have elite resistances to net type net blotch. Four lines out of 69 were chosen for further studies. These four lines: WA 4794 (103 IBON 91), Pompadour, CI 9214, and WPG 8412-9-2-1 were highly diverse and resistant to most of the isolates, and were crossed with Stirling-a highly adaptive but susceptible cultivar. Doubled haploids, F2s, and resistant x resistant crosses were studied against five prevalent isolates. Four genes from WA 4794 (all dominant), three (two dominant and one recessive) from Pompadour, five (two dominant and three recessive) from CI 9214, and two (one dominant and one recessive) from WPG 8412-9-2-1 were identified. In total, 11 different genes were operative against P. teres f. teres isolates. Molecular work is initiated to develop markers which would aid screening of the breeding populations for these resistances.
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Field observations of net blotch epidemics indicated that Tallon barley was quite resistant to infection during later stages of growth despite being susceptible as a seedling. A glasshouse experiment was conducted to determine the effectiveness of this resistance and when it became operative. Three cultivars – Gilbert (very susceptible), Patty (resistant) and Tallon – were inoculated at various stages of growth with conidia of Pyrenophora teres f. teres and the infection response and leaf area diseased, recorded 13 days later. The response of Tallon clearly changed from susceptible to moderately susceptible at growth stage 33. Plants sown two weeks earlier were susceptible and plants sown two weeks later were moderately resistant. The response of the other two cultivars at similar growth stages paralleled their seedling responses. The resistance of Tallon appeared to increase with maturity so that, at its most resistant growth stage, the leaf area diseased was just 10% that of the susceptible, Gilbert. While this resistance appears pathotype specific, this experiment demonstrated very effective APR to net blotch. As most losses to this disease occur during the later stages of plant development, APR offers a valuable source of resistance.
Mapping genes for resistance to net form of net blotch and strip rust in barley (Hordeum vulgare L.)
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A dihaploid mapping population comprising 65 lines was developed between barley parent varieties Tallon and Kaputar and used to construct a genetic linkage map. This map, comprising 195 amplified fragment length polymorphism and 38 simple sequence repeat markers, was used to identify markers linked to the net form of net blotch (Pyrenophora teres f.sp. teres) and to stripe rust (Puccinia striiformis f.sp. hordei) in barley. The population was screened with five pathotypes of net blotch at the seedling stage in the glasshouse and subjected to a natural inoculation in Hermitage, Queensland. Stripe rust screening was conducted at the adult plant stage in Toluca, Mexico. Analyses of the markers were performed using Mapmanager and Qgene software. One region on chromosome 6H was highly significantly associated with resistance to the net blotch (R2 = 79%). This association was consistent for all pathotypes studied. One region on chromosome 5H was found to be highly significantly associated with resistance to stripe rust (R2= 65%). There are a number of very closely linked markers showing strong associations in these regions, and these markers present an opportunity for marker assisted selection of these traits in barley breeding programs.
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World consumption of vegetable oils has increased in recent years because of its application in food, chemical, pharmaceutical and, more recently, energy industry. However, oilseeds, which these oils are extracted, have low viability, affecting the cultivation and productivity of these species. The aim of this study was to analyze the effect of aging on the coordination of catalase (CAT) and ascorbate peroxidase (APX) antioxidant systems in safflower and sunflower. . Therefore, seeds were subjected to accelerated aging for 3, 6 and 9 days and grown in moistened paper towel for 72 hours. Additionally, before accelerated aging, sunflower seeds were pretreated by osmopriming with 10 mM ascorbate (ASC) or 3 amino 1,2,4 triazol (3-AT), a specific inhibitor of CAT activitie. The method of artificial aging used was efficient in both species, because it caused a decrease in germination, seedling development and growth, especially in safflower. The aging caused inhibition of CAT activity for both species and to compensate for such inhibition , sunflower increased mRNA expression of this enzyme , while safflower mobilized over the activity of APX. Analysis of the expression of malate synthase and sugar content demonstrated that sunflower seeds consumes lipid reserves in quiescent state, while the safflower is more dependent on carbohydrate. Pretreatment with 3-AT inhibited CAT activity and stimulated the APX, though with ASC acted reverse on these systems. None of the treatments recovered the physiological decline aging. It is concluded that aging change the oilseeds antioxidant metabolism, despite interspecies variations in response to this process, the depletion of the CAT antioxidant system was common. Because of this we propose that the measurement of CAT activity can be used to identify aging seed lots.
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Peer reviewed