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Comparison of mechanisms controlling uptake and accumulation of 2,4-dichlorophenoxy acetic acid, naphthalene-1-acetic acid, and indole-3-acetic acid in suspension-cultured tobacco cells

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Abstract

Accumulation of radiolabelled naphthalene-1-acetic acid (1-NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), and indole-3-acetic acid (IAA) has been measured in suspension-cultured tobacco (Nicotiana tabacum) cells. In this paper is presented a simple methodology allowing activities of the auxin influx and efflux carriers to be monitored independently by measuring the cellular accumulation of [3H]NAA and [14C]2,4-D. We have shown that 1-NAA enters cells by passive diffusion and has its accumulation level controlled by the efflux carrier. By contrast, 2,4-D uptake is mostly ensured by the influx carrier and this auxin is not secreted by the efflux carrier. Both auxin carriers contribute to IAA accumulation. The kinetic parameters and specificity of each carrier have been determined and new information concerning interactions with naphthylphthalamic acid, pyrenoylbenzoic acid, and naphthalene-2-acetic acid are provided. The relative contributions of diffusion and carrier-mediated influx and efflux to the membrane transport of 2,4-D, 1-NAA, and IAA have been quantified, and the data indicate that plant cells are able to modulate over a large range their auxin content by modifying the activity of each carrier.

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Abbreviations

2,4-D:

2,4-dichlorophenoxyacetic acid

1-NAA:

naphthalene-1-acetic acid

2-NAA:

naphthalene-2-acetic acid

NPA:

N-1-naphthylphthalamic acid

PBA:

2-(1-pyrenoyl)benzoic acid

Vm :

maximum transport capacity of the carrier

References

  • Astle MC, Rubery PH (1983) Carriers for abscisic acid and indole-3-acetic acid in primary roots: their regional localisation and thermodynamic driving forces. Planta 157: 53–63

    Article  CAS  PubMed  Google Scholar 

  • Bean RC, Shepherd WC, Chan H (1968) Permeability of lipid bilayer membranes to organic solutes. J Gen Physiol 52: 495–508

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Benning C (1986) Evidence supporting a model of voltage-dependent uptake of auxin into Cucurbita vesicles. Planta 169: 228–237

    Article  CAS  PubMed  Google Scholar 

  • Davies PJ (1995) The plant hormones: their nature, occurrence, and functions. In: Davies PJ (ed) Plant hormones: physiology, biochemistry and molecular biology, 2nd edn. Kluwer Academic Publishers, Dordrecht, Boston, London, pp 1–12

    Chapter  Google Scholar 

  • Delbarre A, Muller P, Imhoff V, Morgat JL, Barbier-Brygoo H (1994) Uptake, accumulation and metabolism of auxins in tobacco leaf protoplasts. Planta 195: 159–167

    Article  CAS  Google Scholar 

  • Depta H, Rubery PH (1984) A comparative study of carrier participation in the transport of 2,3,5-triiodobenzoic acid, indole-3-acetic acid, and 2,4-dichlorophenoxyacetic acid by Cucurbita pepo L. hypocotyl segments. J Plant Physiol 115: 371–387

    Article  CAS  PubMed  Google Scholar 

  • Depta H, Eisele KH, Hertel R (1983) Specific inhibitors of auxin transport: action on tissue segments and in-vitro binding to membranes from maize coleoptiles. Plant Sci Lett 31: 181–192

    Article  CAS  Google Scholar 

  • Dibb-Fuller JE, Morris DA (1992) Studies on the evolution of auxin carriers and phytotropin receptors: transmembrane auxin transport in unicellular and multicellular Chlorophyta. Planta 186: 219–226

    Article  CAS  PubMed  Google Scholar 

  • Edwards KL, Goldsmith MHM (1980) pH-dependent accumulation of indoleacetic acid by corn coleoptile sections. Planta 147: 457–466

    Article  CAS  PubMed  Google Scholar 

  • Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 15: 148–151

    Google Scholar 

  • Gimmler H, Heilmann B, Demmig B, Hartung W (1981) The permeability coefficients of the plasmalemma and the chloroplast envelope of spinach mesophyll cells for phytohormones. Z Naturforsch 36c: 672–678

    CAS  Google Scholar 

  • Goldsmith MHM (1977) The polar transport of auxin. Annu Rev Plant Physiol 28: 439–478

    Article  CAS  Google Scholar 

  • Hertel R (1987) Auxin transport: binding of auxins and phytotropins to the carriers. Accumulation into and efflux from membrane vesicles. In: Klämbt D (ed) Plant hormone receptors (NATO ASI Series, vol H10). Springer-Verlag, Berlin, Heidelberg, pp 81–92

    Chapter  Google Scholar 

  • Hertel R, Evans ML, Leopold AC, Sell HM (1969) The specificity of the auxin transport system. Planta 85: 238–249

    Article  CAS  PubMed  Google Scholar 

  • Hertel R, Lomax TL, Briggs WR (1983) Auxin transport in membrane vesicles from Cucurbita pepo L. Planta 157: 193–201

    Article  CAS  PubMed  Google Scholar 

  • Heyn A, Hoffmann S, Hertel R (1987) In-vitro auxin transport in membranes vesicles from maize coleoptiles. Planta 172: 285–287

    Article  CAS  PubMed  Google Scholar 

  • Jacobs M, Hertel R (1978) Auxin binding to subcellular fractions from Cucurbita hypocotyls: in vitro evidence for an auxin transport carrier. Planta 142: 1–10

    Article  CAS  PubMed  Google Scholar 

  • Kaldewey H (1984) Transport and other modes of movement of hormones (mainly auxins). In: Scott TK (ed) Encyclopedia of plant physiology, NS, vol 10, Hormonal regulation of development II: the function of hormones from the level of the cell to the whole plant. Springer-Verlag, Berlin, Heidelberg, New York, Tokyo, pp 80–148

    Google Scholar 

  • Katekar GF (1979) Auxins: on the nature of the receptor site and molecular requirements for auxin activity. Phytochemistry 18: 223–233

    Article  CAS  Google Scholar 

  • Liu CM, Xu ZH, Chua NH (1993) Auxin polar transport is essential for the establishment of bilateral symmetry during early plant embryogenesis. Plant Cell 5: 621–630

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lomax TL, Mehlhorn RJ, Briggs WR (1985) Active auxin uptake by zucchini membrane vesicles: quantitation using ESR volume and ΔpH determinations. Proc Natl Acad Sci USA 82: 6541–6545

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lomax TL, Muday GK, Rubery PH (1995) Auxin transport. In: Davies PJ (ed) Plant hormones: physiology, biochemistry and molecular biology, 2nd edn. Kluwer Academic Publishers, Dordrecht, Boston, London, pp 509–530

    Chapter  Google Scholar 

  • Loper MT, Spanswick RM (1991) Auxin transport in suspension-cultured soybean root cells I. Characterization. Plant Physiol 96: 184–191

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Martin HV, Pilet PE (1986) Saturable uptake of indol-3yl-acetic acid by maize roots. Plant Physiol 81: 889–895

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Michalke W, Katekar GF, Geissler AE (1992) Phytotropin-binding sites and auxin transport in Cucurbita pepo: evidence for two recognition sites. Planta 187: 254–260

    Article  CAS  PubMed  Google Scholar 

  • Morris DA, Rubery PH, Jarman J, Sabater M (1991) Effects of inhibitors of protein synthesis on transmembrane auxin transport in Cucurbita pepo L. hypocotyl segments. J Exp Bot 42: 773–783

    Article  CAS  Google Scholar 

  • Muller JF, Goujaud J, Caboche M (1985) Isolation in vitro of naphthalene-acetic acid-tolerant mutants of Nicotiana tabacum, which are impaired in root morphogenesis. Mol Gen Genet 199: 194–200

    Article  CAS  Google Scholar 

  • Okada K, Ueda J, Komaki MK, Bell CJ, Shimura Y (1991) Requirement of the auxin polar transport system in early stages of Arabidopsis floral bud formation. Plant Cell 3: 677–684

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pickard BG (1985a) Roles of hormones, protons and calcium in geotropism. In: Pharis RP, Reid DM (eds) Encyclopedia of plant physiology, NS, vol 11, Hormonal regulation of development III: role of environmental factors. Springer-Verlag, Berlin, Heidelberg, New York, Tokyo, pp 193–281

    Google Scholar 

  • Pickard BG (1985b) Roles of hormones in phototropism. In: Pharis RP, Reid DM (eds) Encyclopedia of plant physiology, NS, vol 11, Hormonal regulation of development HI: role of environmental factors. Springer-Verlag, Berlin, Heidelberg, New York, Tokyo, pp 365–417

    Google Scholar 

  • Raven J (1975) Transport of indoleacetic acid in plant cells in relation to pH and electrical potential gradients, and its significance for polar IAA transport. J New Phytol 74: 163–172

    Article  CAS  Google Scholar 

  • Ray PM, Dohrmann U, Hertel R (1977) Specificity of auxin-binding sites on maize coleoptile membranes as possible receptor sites for auxin action. Plant Physiol 60: 585–591

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rayle D, Ouiralkul R, Hertel R (1969) Effects of auxins on the auxin transport system in coleoptiles. Planta 87: 49–53

    Article  CAS  PubMed  Google Scholar 

  • Rose S, Rubery PH, Bopp M (1983) The mechanism of auxin uptake and accumulation in moss protonemata. Physiol Plant 58: 52–56

    Article  CAS  Google Scholar 

  • Rubery PH (1977) The specificity of carrier-mediated auxin transport by suspension-cultured crown gall cells. Planta 135: 275–283

    Article  CAS  PubMed  Google Scholar 

  • Rubery PH (1978) Hydrogen ion dependence of carrier-mediated auxin uptake by suspension-cultured crown gall cells. Planta 142: 203–206

    Article  CAS  PubMed  Google Scholar 

  • Rubery PH (1987) Auxin transport. In: Davies PJ (ed) Plant hormones and their role in plant growth and development. Martinus Nijhoff, Dordrecht, Boston, Lancaster, pp 341–362

    Chapter  Google Scholar 

  • Rubery PH, Sheldrake AR (1974) Carrier-mediated auxin transport. Planta 118: 101–121

    Article  CAS  PubMed  Google Scholar 

  • Sabater M, Sabater F (1986) Auxin carriers in membranes of lupin hypocotyls. Planta 167: 76–80

    Article  CAS  PubMed  Google Scholar 

  • Sabater M, Rubery PH (1987) Auxin carriers in Cucurbita vesicles III. Specificity, with particular references to 1-naphthylacetic acid. Planta 171: 514–518

    Article  CAS  PubMed  Google Scholar 

  • Schiavone FM (1988) Microamputation of somatic embryos of the domestic carrot reveals apical control of axis elongation and root regeneration. Development 103: 657–664

    Google Scholar 

  • Schiavone FM, Cooke TJ (1987) Unusual patterns of somatic embryogenesis in the domesticated carrot: developmental effects of exogenous auxins and auxin transport inhibitors. Cell Diff 21: 53–62

    Article  CAS  Google Scholar 

  • Sussman MR, Goldsmith MHM (1981) Auxin uptake and action of N-1-naphthylphthalamic acid in corn coleoptiles. Planta 150: 15–25

    Article  Google Scholar 

  • Warren Wilson J, Warren Wilson PM (1993) Mechanisms of auxin regulation of structural and physiological polarity in plants, tissues, cells and embryos. Aust J Plant Physiol 20: 555–571

    Article  Google Scholar 

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In honour of Professor Dieter Klämbt's 65th birthday

The authors thank Drs. A.E. Geissler and G.F. Katekar (CSIRO, Canberra City, Australia) for providing auxin efflux carrier inhibitors CPD, CPP, and PBA, and Dr. H. Barbier-Brygoo (Institut des Sciences Végétales, CNRS, Gif-sur-Yvette, France) for helpful discussions. This work was supported by funds from the Centre National de la Recherche Scientifique (UPR0040).

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Delbarre, A., Muller, P., Imhoff, V. et al. Comparison of mechanisms controlling uptake and accumulation of 2,4-dichlorophenoxy acetic acid, naphthalene-1-acetic acid, and indole-3-acetic acid in suspension-cultured tobacco cells. Planta 198, 532–541 (1996). https://doi.org/10.1007/BF00262639

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  • DOI: https://doi.org/10.1007/BF00262639

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