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71.
Summary The release of intact CU(I)8-thionein from copper-resistant copper-loaded yeast cells, strain X 2180-1Aa, has been shown. This copper(I)-thiolate-rich protein was characterized and compared with the chemical and physicochemical properties of intracellular yeast Cu-thionein. The same molecular mass and stoichiometry of 8 mol copper atoms/mol protein was found. No detectable difference between the Cu-thioneins was seen in luminescence emission, electronic absorption in the ultraviolet region, chiroptical data or amino acid composition. The importance of stable Cu(I)-thiolates in Cu-thionein as a safe vehicle for transporting copper in a non-reactive manner is confirmed. 相似文献
72.
Fusion of cultured dog kidney (MDCK) cells: I. Technique,fate of plasma membranes and of cell nuclei
Ulrich Kersting Heribert Joha Wieland Steigner Birgit Gassner Gerhard Gstraunthaler Walter Pfaller Hans Oberleithner 《The Journal of membrane biology》1989,111(1):37-48
Summary The evaluation of the intracellular signal train and its regulatory function in controlling transepithelial transport with electrophysiological methods often requires intracellular measurements with microelectrodes. However, multiple impalements in epithelial cells are hampered by the small size of the cells. In an attempt to avoid these problems we fused cells of an established cell line, Madin Darby canine kidney cells, originally derived from dog kidney, to giant cells by applying a modified polyethylene glycol method. During trypsin-induced detachment from the ground of the petri dish, individual cells grown in a monolayer incorporate volume and mainly lose basolateral plasma membrane by extrusion. By isovolumetric cell-to-cell fusion, spherical giant cells are formed within 2 hr. During this process a major part of the individual cell plasma membranes is internalized. Over three weeks following cell plasma membrane fusion degradation of single cell nuclei and cell nuclear fusion occurs. We conclude that this experimental approach opens the possibility to investigate ion transport of epithelia in culture by somatic cell genetic techniques. 相似文献
73.
Leaf thionins, a novel class of putative defence factors 总被引:1,自引:0,他引:1
Leaf thionins of barley have been identified as a novel class of highly abundant polypeptides with antifungal activity, which are present in walls and vacuoles of barley leaf cells. Similar thionins occur not only in monocotyledonous but also in various dicotyledonous plants. The leaf thionins of barley are encoded by a complex multigene family, which consists of at least 50–100 members per haploid genome. The toxicity of these thionins for plant pathogenic fungi and the fact that their synthesis can also be triggered by pathogens strongly suggest that leaf thionins are involved in the mechanism of plant defence against microbiol infection. 相似文献
74.
Determination of the pressure in the water-conducting vessels of intactNicotiana rustica L. plants showed that the pressure probe technique gave less-negative values than the Scholander-bomb method. Even though
absolute values of the order of −0.1 MPa could be directly recorded in the xylem by means of the pressure probe, pressures
between zero and atmospheric were also frequently found. The data obtained by the pressure probe for excised leaves showed
that the Scholander bomb apparently did not read the actual tension in the xylem vessles ofNicotiana plants. The possibility that the pressure probe gave false readings was excluded by several experimental controls. In addition,
cavitation and leaks either during the insertion of the microcapillary of the pressure probe, or else during the measurements
were easily recognized when they occurred because of the sudden increase of the absolute xylem tension to that of water vapour
or to atmospheric, respectively. Tension values of the same order could also be measured by means of the pressure probe in
the xylem vessels of pieces of stem cut from leaves and roots under water and clamped at both ends. The magnitude of the absolute
tension depended on the osmolarity of the bathing solution which was adjusted by addition of appropriate concentrations of
polyethylene glycol. Partial and uniform pressurisation of plant tissues or organs, or of entire plants (by means of the Scholander
bomb or of a hyperbaric chamber, respectively) and simultaneous recording of the xylem tension using the pressure probe showed
that a 1∶1 response in xylem pressure only occurred under a few circumstances. A 1∶1 response required that the xylem vessels
were in direct contact with an external water reservoir and/or that the tissue was (pre-)infiltrated with water. Corresponding
pressure-probe measurements in isolated vascular bundles ofPlantago major L. orP. lanceolata L. plants attached to a Hepp-type osmometer indicated that the magnitude of the tension in the xylem vessels was determined
by the external osmotic pressure of the reservoir. These and other experiments, as well as analysis of the data using classical
thermodynamics, indicated that the turgor and the internal osmotic pressure of the accessory cells along the xylem vessels
play an important role in the maintenance of a constant xylem tension. This conclusion is consistent with the cohesion theory.
In agreement with the literature (P.E. Weatherley, 1976, Philos. Trans. R. Soc. London Ser. B23, 435–444; 1982, Encyclopedia of plant physiology, vol. 12B, 79-109), it was found that the tension in the xylem of intact
plants under normal and elevated ambient pressure (as measured with the pressure probe) under quasi-stationary conditions
was independent of the transpiration rate over a large range, indicating that the conductance of the flow path must be flow-dependent. 相似文献
75.
A chemiluminescent assay for hepatitis-B surface antigen is described which used an isoluminol derivative as the label. The assay is precise intra-assay CV, 1.96-2.45%; inter-assay CV, 5.26-8.11% and has a lower detection limit for hepatitis-B surface antigen of 1.3U/I. 相似文献
76.
High mutant frequencies indicated that the wild-type strains of Pichia stipitis are haploid. Sporulation ability of these clones pointed to a homothallic life cycle. Mating was induced by cultivation under
nutritionally poor conditions on malt extract medium. Conjugation was followed immediately by sporulation. However, hybrids
could be rescued by transferring the nascent zygotes to complete medium before meiosis had started. Under rich nutritional
conditions, hybrids were mitotically stable and did not sporulate. The segregation pattern of auxotrophic markers of diploid
zygotes indicated regular meiosis, although asci contained preferentially spore dyads.
Received: 29 February 1996 / Accepted: 29 March 1996 相似文献
77.
Nikolai G. Bukhov Christian Wiese Spidola Neimanis Ulrich Heber 《Photosynthesis research》1996,50(2):181-191
The light-induced induction of components of non-photochemical quenching of chlorophyll fluorescence which are distinguished by different rates of dark relaxation (qNf, rapidly relaxing and qNs, slowly relaxing or not relaxing at all in the presence brief saturating light pulses which interrupt darkness at low frequencies) was studied in leaves of spinach.After dark adaptation of the leaves, a fast relaxing component developed in low light only after a lag phase. Quenching increased towards a maximum with increasing photon flux density. This fast component of quenching was identified as energy-dependent quenching qE. It required formation of an appreciable transthylakoid pH and was insignificant when darkened spinach leaves received 1 s pulses of light every 30 s even though zeaxanthin was formed from violaxanthin under these conditions.Another quenching component termed qNs developed in low light without a lag phase. It was not dependent on a transthylakoid pH gradient, decayed exponentially with a long half time of relaxation and was about 20% of total quenching irrespective of light intensity. When darkened leaves were flashed at frequencies higher than 0.004 Hz with 1 s light pulses, this quenching also appeared. Its extent was very considerable, and it did not require formation of zeaxanthin. Relaxation was accelerated by far-red light, and this acceleration was abolished by NaF.We suggest that qNs is the result of a so-called state transition, in which LHC II moves after its phosphorylation from fluorescent PS II to nonfluorescent PS I. This state transition was capable of decreasing in darkened leaves the potential maximum quantum efficiency of electron flow through Photosystem II by about 20%.Abbreviations PFD
photon flux density
- PS
photosystem 相似文献
78.
79.
Leaves of Kalanchoë daigremontiana Hamet et Perr. at a photon flux density (PFD) above 220 mol·m–2s–1 (400–700 nm) or at leaf temperatures above 27.0 °C showed a rapid loss of rhythmicity, and a more or less pronounced damping-out of the endogenous circadian rhythm of CO2 exchange under continuous illumination. This rhythm was reinitiated after reduction of the PFD by 90–120 mol·m–2·s–1 or reduction of leaf temperature by 3.5–11.0 °C under otherwise unchanged external conditions. The reduction in the magnitude of the external control parameter of the Crassulacean acid metabolism (CAM) rhythm (i.e. PFD or leaf temperature) set the phase of the new rhythm. The maxima of CO2 uptake occurred about 5, 28, 51, 75 h after the reduction. Simulations with a CAM model under comparable conditions showed a similar behaviour. The influence of temperature on the endogenous CAM rhythm observed in K. daigremontiana in vivo could be simulated by incorporating into the model temperature-dependent switch modes for passive efflux of malate from the vacuole to the cytoplasm. Thus, the model indicates that tonoplast function plays an important role in regulation of the endogenous CAM rhythm in K. daigremontiana.Abbreviations CAM
Crassulacean acid metabolism
- PAR
photosynthetically active radiation
- PFD
photon flux density
This work was supported by a grant to F.B. and U.L. from Teilprojekt B5 in the Sonderforschungsbereich 199 of the Deutsche Forschungsgemeinschaft (Bonn, Germany) and by a grant to T. E. E. G. from the Sudienstiftung des deutschen Volkes (Bonn, Germany). Erika Ball is thanked for processing of time-course data for the analysis of Fourier spectra. 相似文献
80.
Atmospheric ammonia (NH3) from various anthropogenic sources has become a serious problem for natural vegetation. Ammonia not only causes changes in plant nitrogen metabolism, but also affects the acid-base balance of plants. Using the pH-sensitive fluorescent dyes pyranine and esculin, cytosolic and vacuolar pH changes were measured in leaves of C3 and C4 plants exposed for brief periods to concentrations of NH3 in air ranging from 1.33 to 8.29 mol NH3 · mol-1 gas (0.94–5.86 mg · m-3). After a lag phase, uptake of NH3 from air at a rate of 200 nmol NH3 · m - 2 leaf area · s- 1 into leaves of Zea mays L. increased pyranine fluorescence indicating cytosolic alkalinisation. The increase was much larger in the dark than in the light. In illuminated leaves of the C3 plant Pelargonium zonale L. and the C4 plants Z. mays and Amaranthus caudatus L., NH3-dependent cytosolic alkalinisation was particularly pronounced when CO2 was supplied at very low levels (16 or 20 mol CO2 · mol- 1 gas, containing 210 mmol O2 · mol- 1 gas). An increase in esculin fluorescence, which was smaller than that of pyranine, was indicative of trapping of some of the NH3 in the vacuoles of leaves of Spinacia oleracea L. and Z. mays. Photosynthesis and transpiration remained unchanged during exposure of illuminated leaves to NH3, yielding an influx of 200 nmol NH3 · m-2 leaf area · s-1 for up to 30 min, the longest exposure time used. Both CO2 and O2 influenced the extent of cytosolic alkalinisation. At 500 mol CO2 · mol-1 gas the cytosolic alkalinisation was suppressed more than at 16 or 20 mol CO2 · mol-1 gas. The suppressing effect of CO2 on the NH3induced alkalinisation was larger in illuminated leaves of the C4 plants Z. mays and A. caudatus than in leaves of the C3 plant P. zonale. A reduction of the O2 concentration from 210 to 10 mmol O2 · mol -1 gas, which inhibits photorespiration, increased the NH3induced cytosolic alkalinisation in C3 plants. Suppression by CO2 or O2 of the alkaline pH shift caused by the dissolution and protonation of NH3 in queous leaf compartments, and possibly by the production of organic compounds synthesised from atmospheric NH3, indicates that NH3 which enters leaves is rapidly assimilated if photosynthesis or photorespiration provide nitrogen acceptor molecules.This work was supported by the Biotechnology and Biological Sciences Research Council and the Deutsche Forschungsgemein-schaft within the framework of the research of Sonderforschun-gsbreich 251 of the University of Würzburg. We are grateful to Dr. B. Wollenweber (The Royal Veterinary and Agricultural University, Denmark) for discussions. 相似文献