Summary
Water uptake ofArabidopsis thaliana protoplasts was measured after transfer into hypo-osmotic conditions. The time-dependent swelling of protoplast populations was monitored by a Coulter counter device. In order to ascertain the contribution of the plasma membrane intrinsic protein 1b (PIP1b) to the membrane's water permeability, protoplasts of five different plant lines that were transformed with a PIP1b antisense construct were compared to controls. The size distribution of 5 independent protoplast preparations provided similar results for control and antisense lines under iso-osmolar conditions. After transfer into hypo-osmotic conditions, a time difference for the swelling of protoplasts from the different sources was observed. The sizes of control protoplasts changed in less than 20 s, which indicates high water influx rates. In contrast, the protoplast populations obtained from 5 different antisense plants took about 75 s to reach a steady-state cell size distribution. The difference in time by a factor of about 3 confirms the significance of the aquaporin PIP1b for the water permeability of plant plasma membranes and the cellular water transport.
Similar content being viewed by others
References
Becker D, Kemper E, Schell J, Masterson R (1992) New plant binary vectors with selectable markers located proximal to the left T-DNA border. Plant Mol Biol 20: 1195–1197
Bevan M (1984) BinaryAgrobacterium vectors for plant transformation. Nucleic Acids Res 12: 8711–8721
Ditta G, Stanfield S, Corbin D, Helinsky D (1980) Broad host range DNA cloning system for Gram-negative bacteria: construction of a gene bank fromRhizobium melilotis. Proc Natl Acad Sci USA 77: 7347–7351
Grote K, von Trzebiatovski P, Kaldenhoff R (1998) RNA levels of plasma membrane aquaporins inArabidopsis thaliana. Protoplasma 204: 139–144
Johnson KD, Höfte H. Chrispeels MJ (1990) An intrinsic tonoplast protein of protein storage vacuoles in seeds is structurally related to a bacterial solute transporter (GlpF). Plant Cell 2: 525–532
Kaldenhoff R, Grote K, Zhu JJ, Zimmermann U (1998) Significance of plasmalemma aquaporins for water-transport inArabidopsis thaliana. Plant J 14: 121–128
—, Kölling A, Meyers J, Karman U, Ruppel G, Richter G (1995) The blue light-responsiveAthH2 gene ofArabidopsis thaliana is primarily expressed in expanding as well as in differentiating cells and encodes a putative channel protein of the plasmalemma. Plant J 7: 87–95
— — —, Richter G (1993) A novel blue light- and abscisic acid-inducible gene ofArabidopsis thaliana encoding an intrinsic membrane protein. Plant Mol Biol 23: 1187–1198
Kammerloher W, Fischer U, Piechottka GP, Schaffner AR (1994) Water channels in the plant plasma membrane cloned by immunoselection from a mammalian expression system. Plant J 6: 187–199
Ludevid D, Höfte H, Himelblau E, Chrispeels MJ (1992) The expression pattern of the tonoplast intrinsic protein γ-TIP inArabidopsis thaliana is correlated with cell enlargement. Plant Physiol 100: 1633–1639
Maurel C (1997) Aquaporins and water permeability of plant membranes. Annu Rev Plant Physiol Plant Mol Biol 48: 399–429
Preston GM, Carroll TP, Guggino WB, Agre P (1992) Appearance of water channels inXenopus oocytes expressing red cell CHIP28 protein. Science 256: 385–387
Reizer J, Reizer A, Saier MH Jr (1993) The MIP family of intergral membrane channel proteins: sequence comparisons, evolutionary relationships, reconstructed pathway of evolution, and proposed functional differentiation of the two repeated halves of the proteins. Crit Rev Biochem Mol Biol 28: 235–257
Valvekens D, van Montagu M, van Lijsebettens M (1988)Agrobacterium tumefaciens-mediated transformation ofArabidopsis thaliana root expiants by using kanamycin selection. Proc Natl Acad Sci USA 85: 5536–5540
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Grote, K., Gimmler, H. & Kaldenhoff, R. Coulter counter cell size determination of protoplasts fromArabidopsis thaliana PIP1b aquaporin antisense lines under iso- and hypo-osmotic conditions. Protoplasma 210, 31–35 (1999). https://doi.org/10.1007/BF01314952
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01314952