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This study was undertaken to determine which of the two NO3? fluxes (influx or efflux) across plasma membranes of root cells is the target of those amino acids which have been shown to inhibit net NO3? uptake (Muller & Touraine 1992, Journal of Experimental Botany 43 , 617–623). Parallel experiments were performed to mea-sure either the time course of 15NO3? release from roots of soybean seedlings previously labelled with this isotope into non-labelled solution, or the time course of 15N accumulation from labelled 15NO3? solution in non-labelled seedlings. Focusing on the fate of 15NO3? in the cytoplasmic compartment, a model is developed to describe the time courses of the accumulation and release of tracer across the plasma membranes of root cells. Both time courses can be described by the sum of an exponential plus a linear term. In our material, the linear part of the accumulation time course is obscured by the NO3? fluxes exiting the cytoplasm, and the curve thus appears to be quasilinear over several minutes. However, we show that the use of the net tracer accumulation rate during this time period as an estimate of NO3? influx does not provide accurate estimates of influx and efflux. By contrast, 15NO3? efflux analysis permits calculation of the unidirectional fluxes across plasma membranes of root cells and the kinetic parameters of the cytoplasmic NO3? pool. Under our experimental conditions, efflux accounted for 30 to 50% of influx, and the cytoplasmic NO3? content was found to be in the 70–400nmol g?1 fw range. Using this methodology, the effect of amino acid accumulation on unidirectional fluxes of nitrate was then examined. Pretreatments of the seedlings with an amino acid which has been shown to inhibit net NO3? uptake led to concomitant decreases in net accumulation rates of 15NO3? and of reduced 15N in roots and total 15N in cotyledons. NO3? influx was markedly inhibited by these treatments, while NO3? efflux remained essentially unaffected, or even decreased. It is concluded that the target of the regulation of NO3? uptake by phloemtranslocated amino acids is the influx system. 相似文献
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Differential Stimulation of PEP-carboxylation in Guard Cells and Mesophyll Cells by Ammonium or Fusicoccin 总被引:1,自引:0,他引:1
Dark CO2-fixation in guard cells of Vicia faba was much moresensitive to ammonium than in mesophyll cells. Addition of ammonium(5.0 mol m3; pH0 7.6) caused up to a 7-fold increasein dark CO2-fixation rates in guard cell protoplasts (GCP),whereas in leaf slices, mesophyll cells, and mesophyll protoplaststhe increase was only about 1.4-fold. In both cell or tissuetypes, total CO2-fixation rates were higher in the light (212-foldhigher in GCP and 28-fold in mesophyll); these rates were onlyslightly changed by ammonium treatment. However, separationof 14C-labelled products after fixation of CO2 in the lightby GCP revealed a large ammonium-induced shift in carbon flowfrom starch and sugars to typical products of C4-metabolism(mainly malate and aspartate). In contrast, in mesophyll cellsamino acid and malate labelling was only moderately increasedby ammonium at the expense of sucrose. The data suggest thatin vivo ammonium might facilitate stomatal opening and/or delaystomatal closing through an increased production of organicacids. Key words: PEP-carboxylation, guard cell protoplasts, ammonium, fusicoccin 相似文献
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LANG JUDITH C.; LASKER HOWARD R.; GLADFELTER ELIZABETH H.; HALLOCK PAMELA; JAAP WALTER C.; LOSADA FREDDY J.; MULLER ROBERT G. 《Integrative and comparative biology》1992,32(6):696-706
Western Atlantic coral reefs were differentially affected bya mass bleaching (discoloration) event in 1987. We periodicallyassessed the "appearance" of zooxanthellate organisms betweenDecember 1987 and June 1988 at nine conspicuously affected sitesin the Bahamas, Florida, St. Croix, and Venezuela, using a standardizedpoint-count technique. Three to four months after the localinitiation of the event, the "bleached" state was still presentin one to three of the most abundant reef coral taxa and ina few of the less common species (n = 5 sites). "Recovery" occurredsomewhat faster at shallower depths, at least in the Bahamasand Florida. Scleractinian corals which were "prolonged bleachers"had foliaceous or massive, rather than branching, morphologies."Bleached" points disappeared from the point counts after $6to $8 months. Long-term field data on spatial and temporal variability inthe dynamics of zooxanthellate organisms would help us to understandthe ecological consequences of bleaching. More generally, weneed to distinguish anthropogenic changes in the structure andfunctioning of reef ecosystems from those which occur naturally.Point-count techniques are well suited for collaborative studiesinvolving rapid quantification of coloration states and healthin reef corals. 相似文献
15.
Interaction between atmospheric and pedospheric nitrogen nutrition in spruce (Picea abies L. Karst) seedlings 总被引:4,自引:2,他引:4
The effect of NO2 fumigation on root N uptake and metabolism was investigated in 3-month-old spruce (Picea abics L. Karst) seedlings. In a first experiment, the contribution of NO2 to the plant N budget was measured during a 48 h fumigation with 100mm3m?3 NO2. Plants were pre-treated with various nutrient solutions containing NO2 and NH4+, NO3? only or no nitrogen source for 1 week prior to the beginning of fumigation. Absence of NH4+ in the solution for 6d led to an increased capacity for NO3? uptake, whereas the absence of both ions caused a decrease in the plant N concentration, with no change in NO3? uptake. In fumigated plants, NO2 uptake accounted for 20–40% of NO3? uptake. Root NO3? uptake in plants supplied with NH4+plus NO3? solutions was decreased by NO2 fumigation, whereas it was not significantly altered in the other treatments. In a second experiment, spruce seedlings were grown on a solution containing both NO2 and NH4+ and were fumigated or not with 100mm3m?3 NO2 for 7 weeks. Fumigated plants accumulated less dry matter, especially in the roots. Fluxes of the two N species were estimated from their accumulations in shoots and roots, xylem exudate analysis and 15N labelling. Root NH4+ uptake was approximately three times higher than NO3? uptake. Nitrogen dioxide uptake represented 10–15% of the total N budget of the plants. In control plants, N assimilation occurred mainly in the roots and organic nitrogen was the main form of N transported to the shoot. Phloem transport of organic nitrogen accounted for 17% of its xylem transport. In fumigated plants, neither NO3? nor NH4+ accumulated in the shoot, showing that all the absorbed NO2 was assimilated. Root NO3? reduction was reduced whereas organic nitrogen transport in the phloem increased by a factor of 3 in NO2-fimugated as compared with control plants. The significance of the results for the regulation of whole-plant N utilization is discussed. 相似文献
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MARTIN CLÉMENT CAROLINE CHAMBERLAND JACQUELINE PÉRODIN RICHARD LEDUC GAÉTAN GUILLEMETTE EMANUEL ESCHER 《Journal of receptor and signal transduction research》2013,33(5-6):417-433
Several models of activation mechanisms were proposed for G protein-coupled receptors (GPCRs), yet no direct methods exist for their elucidation. The availability of constitutively active mutants has given an opportunity to study active receptor conformations within acceptable limits using models such as the angiotensin II type 1 (AT1)1 receptor mutant N111G-hAT1 which displays an important constitutive activity. Recently, by using methionine proximity assay, we showed for the hAT1 receptor that TMD III, VI, and VII form the ligand-binding pocket of the C-terminal amino acid of an antagonistic AngII analogue. In the present contribution, we investigated whether the same residues would also constitute the ligand-binding contacts in constitutively activated mutant (CAM) receptors. For this purpose, the same Met mutagenesis strategy was carried out on the N111G double mutants. Analysis of 43 receptors mutants in the N111G-hAT1 series, photolabeled and CNBr digested, showed that there were only subtle structural changes between the wt-receptor and its constitutively active form. 相似文献
19.
CLARE J. HOPKINS NOEL O. I. COGAN MELANIE HAND ERICA JEWELL JATINDER KAUR XI LI GERALDINE A. C. LIM ALISON E. LING CHRISTOPHER LOVE HAYLEY MOUNTFORD MARIJA TODOROVIC MEGAN VARDY GERMAN C. SPANGENBERG DAVID EDWARDS JACQUELINE BATLEY 《Molecular ecology resources》2007,7(4):697-700
The availability of expressed sequence data derived from gene discovery programs enables mining for simple sequence repeats (SSR), providing useful genetic markers for crop improvement. These markers are inexpensive, require minimal labour to produce and can frequently be associated with functionally annotated genes. This study presents the development and characterization of 16 expressed sequence tags (EST)‐SSR markers from Brassica juncea and their cross‐amplification across Brassica species. Sixteen primer pairs were assessed for polymorphism in all genomes of the diploid and amphidiploid Brassica species. The markers show reliable amplification, considerable polymorphism and high transferability across species, demonstrating the utility of EST‐SSRs for genetic analysis of brassicas. 相似文献
20.
JACQUELINE BATLEY CLARE J. HOPKINS NOEL O. I. COGAN MELANIE HAND ERICA JEWELL JATINDER KAUR SUKHJIWAN KAUR XI LI ALISON E. LING CHRISTOPHER LOVE HAYLEY MOUNTFORD MARIJA TODOROVIC MEGAN VARDY MARZENA WALKIEWICZ GERMAN C. SPANGENBERG DAVID EDWARDS 《Molecular ecology resources》2007,7(5):886-889
The availability of expressed sequence data derived from gene discovery programs enables mining for simple sequence repeats (SSR), providing useful genetic markers for crop improvement. These markers are inexpensive, require minimal labour to produce and can frequently be associated with functionally annotated genes. This study presents the development and characterization of 24 expressed sequence tags (EST)‐SSR markers from Brassica napus and their cross‐amplification across Brassica species. The markers show reliable amplification, genome specificity and considerable polymorphism, demonstrating the utility of EST‐SSRs for genetic analysis of wild Brassica populations and commercial Brassica germplasm. 相似文献