Histochemical Evidence for the Occurrence of Oligomycin-sensitive Plasmalemma ATPase in Corn Roots

Malone, Carl P.; Burke, John J.; Hanson, James A.

doi:10.1104/pp.60.6.916

Cited by 35 publications

(8 citation statements)

References 15 publications

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“…The inhibition increases with increasing oligomycin concentration, and the oligomycin concentration giving 50%o inhibition is about 6 (6) show that oligomycin inhibits Cl-influx more than K+ influx in excised barley roots. Lin and Hanson (7) showed that oligomycin inhibits phosphate influx in corn roots to a greater extent than can be accounted for by its effect on ATP levels, and Malone et al (8) …”

Section: Resultsmentioning

confidence: 99%

ATP Levels and their Effects on Plasmalemma Influxes of Potassium Chloride in Red Beet

Petraglia¹,

Poole²

1980

Plant Physiol.

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Tissue ATP concentrations in slices of red beet increase progressively with time for up to 7 days after cutting the root. ATP The manner in which metabolic energy is supplied to ion transport processes in plant cells is not well established. It is generally agreed that cation influx can be coupled to ATP hydrolysis (13), although the manner of coupling, and the relationship of cation influx to electrogenic H+ efflux, remains in doubt. In the present study, the dependence of K+ influx on ATP concentration has been investigated in more detail. We show in another report (14) that the present relationship between influx and ATP concentration is different from that observed for the electrogenic pump activity.With regard to anion influx (13), there is some evidence for its dependence on ATP but contrary claims have also been made, especially for storage tissues (2, 11). The present study shows that in storage tissue of red beet Cl-influx shows a dependence on ATP concentration similar to that observed for K+ influx. Evidence is also presented that, in addition to its effect on ATP levels, oligomycin inhibits the Cl-pump at the plasmalemma. (1) with modifications as indicated below. One g beet disks frozen in liquid N2 were disrupted in 40 ml ice-cold 0.4 N HC104 + 1 mm EDTA disodium salt, in a Virtis homogenizer for 1 min. One-half of the homogenate was taken for isotope counting. This was decolorized with activated charcoal (2.5 ml of a 10%7b suspension in water), and centrifuged 10 min at 15,000g. Eight-ml samples of supernatant + 10 ml Aquasol (New England Nuclear) were counted in a scintillation counter, and then recounted for 36C1 after decay of the 42K. MATERIALS AND METHODSThe remainder of the tissue homogenate was taken for ATP assays after centrifugation at 10,000g for 15 min to sediment debris. Immediately before the ATP assays, 5 ml ofthe supernatant were added to 5 ml 40 mm glycylglycine buffer (pH 7.4) and the mixture was adjusted to pH 7.4 with 1 M KOH + 0.1 M KHCO3. The extract was then centrifuged at 5,000g for 5 min to remove KC104 and ATP assays were performed without delay. The luciferin-luciferase ATP assays were performed without delay. The luciferin-luciferase ATP assays were performed as described by Schram (15) with a few modifications. The content of one vial of firefly extract (Sigma FLE-50) was dissolved in 5 ml cold distilled H20. This reconstitution of the firefly extract gives a final concentration of 50 mm potassium arsenate and 20 mm MgSO4 (pH 7.4). This enzyme solution was agitated occasionally and incubated on ice for 2-5 h. When ready for use, the enzyme solution was centrifuged 10 min at 5,000g. The supernatant was either used immediately in the ATP assay or stored frozen for future use.To scintillation vials containing 700 Id glycylglycine buffer were added 100 tA of the enzyme solution prepared as described above.

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Section: Resultsmentioning

confidence: 99%

ATP Levels and their Effects on Plasmalemma Influxes of Potassium Chloride in Red Beet

Petraglia¹,

Poole²

1980

Plant Physiol.

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“…-ATPase activity analysis, sections were cut from the same leaves that were used for X-ray microanalysis. Pb(NO 3 ) 2 staining was used to indicate ATPase activity as described by Malone et al (1977) and Jian et al (2000) with some modifications. The histochemical localization of ATPase activity is based on the reaction of Pb 2?…”

Section: Cytochemical Analysis Of Hmentioning

confidence: 99%

Effect of NaCl on leaf H+-ATPase and the relevance to salt tolerance in two contrasting poplar species

Deng

et al. 2010

Trees

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During a 30-day period of increasing salinity, we examined the effects of NaCl on leaf H? -ATPase and salinity tolerance in 1-year-old plants of Populus euphratica Oliv. (salt resistant) and P. popularis 35-44 (P. popularis) (salt sensitive). Electron probe X-ray microanalysis of leaf mesophyll revealed that P. euphratica had a higher ability to retain lower NaCl concentrations in the cytoplasm, as compared to P. popularis. The sustained activity of H ? pumps (by cytochemical staining) in salinised P. euphratica suggests a role in energising salt transport through the plasma membrane (PM) and tonoplast. Saltinduced alterations of leaf respiration, ATP content and expression of PM H ? -ATPase were compared between the two species. Results show that P. euphratica retained a constant respiratory rate, ATP production and protein abundance of PM H ? -ATPase (by Western blotting) in salt-stressed plants. P. euphratica was able to maintain a comparatively high capacity of ATP hydrolysis and H ? pumping during prolonged salt exposure. By contrast, the activity and expression of PM H ? -ATPase were markedly decreased in P. popularis leaves in response to salt stress. Furthermore, NaCl-stressed P. popularis plants showed a marked decline of respiration (70%) and ATP production (66%) on day 30. We conclude that the inability of P. popularis to transport salt to the apoplast and vacuole was partly due to the decreased activity of H ? pumps. As a consequence, cytosolic ion concentrations were observed to be comparatively high for an extended period of time, so that cell metabolism, in particular respiration, was disrupted in P. popularis leaves.

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“…Available evidence indicates that at low ionic concentrations in the rhizosphere, mobile ions are absorbed predominantly by cells of the epidermis or outer cortex at the root periphery (14,20,31, and references therein). The radial pathway for N03-movement in the root thus likely would involve uptake by epidermal or outer cortical cells at the root periphery, symplastic transport through the cortex and stele, and release from the symplasm into the xylem at xylem parenchyma cells (6,9,16,27).…”

mentioning

confidence: 99%

Intercellular Localization of Nitrate Reductase in Roots

Rufty¹,

Thomas²,

Remmler³

et al. 1986

Plant Physiol.

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Experiments were conducted with segments of corn roots to investigate whether nitrate reductase (NR) is compartmentalized in particular groups of cells that collectively form the root symplastic pathway. A microsurgical technique was used to separate cells of the epidermis, of the cortex, and of the stele. The presence of NR was determined using in ritro and enzyme-linked immunosorbent assays. In roots exposed to 0.2 millimolar N03-for 20 hours, NR was detected almost exclusively in epidermal cells, even though substantial amounts of NO3-likely were being transported through cortical and steler cells during transit to the vascular system. Although NR was present in all cell groups of roots exposed to 20.0 millimolar NO3-, the majority of the NR still was contained in epidermal cells. The results are consistent with previous observations indicating that limited reduction of endogenous NO3-occurs during uptake and reduction of exogenous NO3-. Several mechanisms are advanced to account for the restricted capacity of cortical and stelar cells to induce NR and reduce NO3-. It is postulated that (a) the biochemical system involved in the induction of NR in the cortex and stele is relatively insensitive to the presence of NO3-, (b) the receptor for the NR induction response and the NR protein are associated with cell plasmalemmae and little NO3-is taken up by cells of the cortex and stele, and/or (c) N03-is compartmentalized during transport through the symplasm, which limits exposure for induction of NR and NO3-reduction.Experiments with wheat and corn seedlings using 15N03-have revealed that NO3-reduction in roots may be localized in a specific cellular compartment (1,17,18). Following induction of the N03 transport and reduction systems, exogenously supplied NO3-being taken up by the root was readily reduced and translocated to the xylem. In contrast, endogenous NO3-which had been previously accumulated in the tissue was readily translocated or effluxed to the external solution, but minimal amounts were reduced.

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Histochemical Evidence for the Occurrence of Oligomycin-sensitive Plasmalemma ATPase in Corn Roots

Cited by 35 publications

References 15 publications

ATP Levels and their Effects on Plasmalemma Influxes of Potassium Chloride in Red Beet

ATP Levels and their Effects on Plasmalemma Influxes of Potassium Chloride in Red Beet

Effect of NaCl on leaf H+-ATPase and the relevance to salt tolerance in two contrasting poplar species

Intercellular Localization of Nitrate Reductase in Roots

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