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Osmotic water permeability of plasma and vacuolar membranes in protoplasts I. High osmotic water permeability in radish (Raphanus sativus) root cells as measured by a new method

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An Erratum to this article was published on 24 February 2007

Abstract

Intra- and transcellular water movements in plants are regulated by the water permeability of the plasma membrane (PM) and vacuolar membrane (VM) in plant cells. In the present study, we investigated the osmotic water permeability of both PM (P f1) and VM (P f2), as well as the bulk osmotic water permeability of a protoplast (P f(bulk)) isolated from radish (Raphanus sativus) roots. The values of P f(bulk) and P f2 were determined from the swelling/shrinking rate of protoplasts and isolated vacuoles under hypo- or hypertonic conditions. In order to minimize the effect of unstirred layer, we monitored dropping or rising protoplasts (vacuoles) in sorbitol solutions as they swelled or shrunk. P f1 was calculated from P f(bulk) and P f2 by using the ‘three-compartment model’, which describes the theoretical relationship between P f1, P f2 and P f(bulk) (Kuwagata and Murai-Hatano in J Plant Res, 2007). The time-dependent changes in the volume of protoplasts and isolated vacuoles fitted well to the theoretical curves, and solute permeation of PM and VM was able to be neglected for measuring the osmotic water permeability. High osmotic water permeability of more than 500 μm s−1, indicating high activity of aquaporins (water channels), was observed in both PM and VM in radish root cells. This method has the advantage that P f1 and P f2 can be measured accurately in individual higher plant cells.

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Abbreviations

C 0 :

extracellular solute concentration (mol m−3)

C 1 :

solute concentration in cytoplasmic space (mol m−3)

C 2 :

solute concentration in vacuolar space (mol m−3). (The second subscripts “0” and “+” in (C 0, C 1, C 2) represent initial and final states of swelling or shrinking)

L P1 :

hydraulic conductivity of PM

L P2 :

hydraulic conductivity of VM

L P(bulk) :

bulk hydraulic conductivity of a protoplast

P f(bulk) :

bulk osmotic water permeability of a protoplast

P f(bulk0) :

P f(bulk) at t = 0 (in the case of constant S × P f )

P f1 :

osmotic water permeability of PM.

P f2 :

osmotic water permeability of VM. (It is noted that P f (or L P ) in the text represents P f1, P f2, or P f(bulk) (or L P1, L P2, or L P(bulk)). Superscript ‘en’ or ‘ex’ indicates a P f (or L P ) value was determined by an endosmotic or exosmotic process, respectively)

r 10 :

initial radius of a protoplast

r 20 :

initial radius of a vacuole

t (obs)0.5 :

half-time (defined as the time at which half of the total change in protoplast volume is completed)

t′ *0.5 :

dimensionless half-time for the two-compartment model in the case of constant P f

t′ s*0.5 :

dimensionless half-time for the two-compartment model in the case of constant S × P f

V 1 :

volumes of a protoplast. (The second subscripts “0” and “+” in V 1 represent initial and final states of swelling or shrinking)

α 0 :

r 20/r 10

β 10 :

ratio of initial and final equilibrium volumes of a protoplast (= V 1+/V 10)

PM:

Plasma membrane

S:

Surface area of a membrane

VM:

Vacuolar membrane

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Acknowledgments

We are indebted to Dr. Shizuo Yoshida (Hokkaido University), Masayoshi Maeshima (Nagoya University), and Hiroshi Nonami (Ehime University) for stimulating our interest in water movement across plant membranes. We are grateful to Dr. Hiroshi Shono and Dr. Matsuo Uemura of Iwate University, Dr. Yasuo Shioya of the National Institute of Livestock and Grassland Science, Dr. Takahiro Hamasaki of the National Agricultural Research Center for the Hokkaido Region, Junko Sakurai and Masumi Okada of the National Agricultural Research Center for the Tohoku Region for their helpful advice on measuring osmotic water permeability. We wish to thank Dr. Joe Wolfe of the University of New South Wales, Sydney, for valuable comments about the physical properties of cellular membranes. We would like to thank Chihaya Nakagawara (Iwate University) and Katsuko Takasugi (National Agricultural Research Center for the Tohoku Region) for providing technical assistance. We also thank Tamito Sakurai for his advice about statistical analysis. We are grateful to the anonymous reviewers for their helpful comments and suggestions. This work was supported by Grants-in-Aid from the Ministry of Education, Sports, Culture, Science and Technology of Japan (Nos.16780181 and 18380151).

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Authors and Affiliations

  1. National Agricultural Research Center for the Tohoku Region, Morioka, 020-0198, Japan

    Mari Murai-Hatano

  2. National Institute for Agro-environmental Sciences, Tsukuba, 305-8604, Japan

    Tsuneo Kuwagata

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  1. Mari Murai-Hatano
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  2. Tsuneo Kuwagata
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Correspondence to Mari Murai-Hatano.

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Mari Murai-Hatano and Tsuneo Kuwagata contributed equally to the paper.

An erratum to this article is available at http://dx.doi.org/10.1007/s10265-007-0072-5.

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Murai-Hatano, M., Kuwagata, T. Osmotic water permeability of plasma and vacuolar membranes in protoplasts I. High osmotic water permeability in radish (Raphanus sativus) root cells as measured by a new method. J Plant Res 120, 175–189 (2007). https://doi.org/10.1007/s10265-006-0035-2

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