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Tonoplast of Beta vulgaris L. contains detergent-resistant membrane microdomains

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Abstract

The experiments conducted on tonoplast of Beta vulgaris L. roots were performed to identify detergent-resistant lipid–protein microdomains (DRMs, interpreted as lipid rafts).The presence of DRMs can be found when dynamic clustering of sphingolipids, sterols, saturated fatty acids is registered, and the insolubility of these microdomains in nonionic detergents at low temperatures is proven. The elucidation of tonoplast microdomains has been based on results obtained with the aid of high-speed centrifuging in the sucrose gradient. The experiments have shown that tonoplast microdomains are rich in sphingolipids, free sterols and saturated fatty acids (such a lipid content is also typical of lipid–protein microdomains of other membranes), while only few phospholipids are present in tonoplast microdomains. The presence of microdomains has been confirmed by fluorescence and confocal microscopy using filipin and Laurdan as fluorescent probes. The experiments with Laurdan have shown that tonoplast microdomains are characterized by a high order compared to characteristics of the rest of the tonoplast. Thus, the presence of detergent-resistant lipid–protein microdomains in the tonoplast has been demonstrated.

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Abbreviations

DBI:

Double bond index

DMSO:

Dimethyl sulfoxide

DRMs:

Detergent-resistant microdomains

EDTA:

Ethylenediaminetetraacetic acid

GP:

Generalized polarization

IMGPs:

Intra-membrane globular particles

PMSF:

Phenylmethanesulfonyl fluoride

TLC:

Thin layer chromatography

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Acknowledgments

The authors are grateful to the colleagues of Siberian Institute of Plant Physiology and Biochemistry, Siberian Branch, Russian Academy of Sciences, namely, Dr. Aleksey V. Stepanov for his assistance in investigations on the fluorescence microscope, to Dr. Lubov V. Dudareva for her assistance in fatty acid analysis, to Mrs. Lidia A. Sitneva and Mrs. Tatiana I. Lapteva for their help in the processes of tonoplast isolation and to Mr. Viktor A. Galichenko for technical assistance in centrifuging.

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Authors and Affiliations

  1. Siberian Institute of Plant Physiology and Biochemistry, Siberian Branch, Russian Academy of Sciences, 132, Lermontov St., Irkutsk, 664033, Russia

    Natalia V. Ozolina, Irina S. Nesterkina, Ekaterina V. Kolesnikova, Ryurik K. Salyaev & Vadim N. Nurminsky

  2. National Research Irkutsk State Technical University, 83, Lermontov St., Irkutsk, 664074, Russia

    Vadim N. Nurminsky

  3. Institute of Laser Physics (Irkutsk Department), Siberian Branch, Russian Academy of Sciences, 130a, Lermontov St., Irkutsk, 664033, Russia

    Alexander L. Rakevich & Evgueni F. Martynovich

  4. Presidium of Irkutsk Scientific Center, Siberian Branch, Russian Academy of Sciences, 134, Lermontov St., Irkutsk, 664033, Russia

    Michael Yu. Chernyshov

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  1. Natalia V. Ozolina
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  2. Irina S. Nesterkina
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  8. Michael Yu. Chernyshov
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Correspondence to Natalia V. Ozolina.

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Ozolina, N.V., Nesterkina, I.S., Kolesnikova, E.V. et al. Tonoplast of Beta vulgaris L. contains detergent-resistant membrane microdomains. Planta 237, 859–871 (2013). https://doi.org/10.1007/s00425-012-1800-1

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