ION-TRANSPORT IN ISOLATED PROTOPLASTS FROM TOBACCO SUSPENSION CELLS .1. GENERAL CHARACTERISTICS

被引:51
作者
METTLER, IJ [1 ]
LEONARD, RT [1 ]
机构
[1] UNIV CALIFORNIA RIVERSIDE, DEPT BOT & PLANT SCI, RIVERSIDE, CA 92521 USA
关键词
D O I
10.1104/pp.63.1.183
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
An investigation was conducted into the feasibility of using enzymically isolated protoplasts from suspension-cultured cells of Nicotiana glutinosa L. to study ion transport. Transport of K+ (86Rb), 36Cl-, H232PO4- and 45Ca2+ from 1 mM salt solutions was determined after separation of intact protoplasts from nonabsorbed tracers by centrifugation through a Ficoll step gradient. Influx of K+, Cl- and H2PO4- measured over a 30 min period was reduced (up to 99%) by respiratory inhibitors such as 5 .mu.g/ml oligomycin, 0.1 mM dinitrophenol, 0.1 mM cyanide, or N2 gas. In contrast, Ca2+ influx was not tightly coupled to respiratory energy production. The influx of K+ was highest between pH 6.5 and 7.5 whereas the influx of H2PO4- and Cl- was greatest between pH 4.5 and 5.5. Influx of K+ and Cl- was maximal at 35 and 45.degree.C, respectively, and was almost completely inhibited below 10.degree. C. Fusicoccin (0.01 mM) stimulated K+ influx by more than 200% but had no effect on the influx of either Cl- or H2PO4-. Apparent H+ efflux, as measured by decrease in solution pH, was enhanced by K+, stimulated further by 0.01 mM fusicoccin, and inhibited by 0.1 mM dinitrophenol or 5 .mu.g/ml oligomycin. The measured ionic fluxes into protoplasts were similar to those obtained with intact cultured cells. Apparently enzymic removal of the cell wall produced no significant alteration in the transport properties of the protoplast and it is probably feasible to use isolated protoplasts for studies on ion transport.
引用
收藏
页码:183 / 190
页数:8
相关论文
共 25 条
  • [1] COVALENT LABELING OF MEMBRANES
    CARRAWAY, KL
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA, 1975, 415 (04) : 379 - 410
  • [2] RAPID STIMULATION OF K+-H+ EXCHANGE BY A PLANT-GROWTH HORMONE
    CLELAND, RE
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1976, 69 (02) : 333 - 338
  • [3] Cocking E C, 1977, Int Rev Cytol, V48, P323, DOI 10.1016/S0074-7696(08)61748-9
  • [4] CHARACTERISTICS OF FUSICOCCIN-INDUCED CHANGES OF TRANSMEMBRANE POTENTIAL AND ION UPTAKE IN MAIZE ROOT SEGMENTS
    COCUCCI, M
    MARRE, E
    BALLARINDENTI, A
    SCACCHI, A
    [J]. PLANT SCIENCE LETTERS, 1976, 6 (03): : 143 - 156
  • [5] MECHANISMS OF ION-TRANSPORT THROUGH PLANT-CELL MEMBRANES
    EPSTEIN, E
    [J]. INTERNATIONAL REVIEW OF CYTOLOGY-A SURVEY OF CELL BIOLOGY, 1973, 34 : 123 - 168
  • [6] GALBRAITH DW, 1977, J CELL SCI, V24, P295
  • [7] MEMBRANE-TRANSPORT OF SUGARS AND AMINO-ACIDS IN ISOLATED PROTOPLASTS
    GUY, M
    REINHOLD, L
    LATIES, GG
    [J]. PLANT PHYSIOLOGY, 1978, 61 (04) : 593 - 596
  • [8] Hodges T. K., 1973, Advances in Agronomy, V25, P163
  • [9] FUSICOCCIN-INDUCED, K+-STIMULATED PROTON SECRETION AND ACID-INDUCED GROWTH OF APICAL ROOT SEGMENTS
    LADO, P
    DEMICHELIS, MI
    CERANA, R
    MARRE, E
    [J]. PLANT SCIENCE LETTERS, 1976, 6 (01): : 5 - 20
  • [10] EFFECT OF LANTHANUM ON ION ABSORPTION IN CORN ROOTS
    LEONARD, RT
    NAGAHASHI, G
    THOMSON, WW
    [J]. PLANT PHYSIOLOGY, 1975, 55 (03) : 542 - 546