Abstract
For the genetic potential of model systems such as Arabidopsis thaliana to be most effectively used to understand drought resistance, reliable and rapid protocols are needed for laboratory study of phenotypes relevant to stress responses in the field. Osmotic adjustment, the amount of additional solutes accumulated by plants under water stress, is often measured in drought physiology studies and requires quantification of both relative water content and solute content (osmotic potential) of the plant tissue. Water stress also elicits high levels of proline accumulation. Protocols are presented here to measure both of these parameters in Arabidopsis seedlings that have been exposed to controlled water stress treatments using polyethylene glycol–agar plates. For the ninhydrin-based assay of proline, a protocol for performing the assay in 96-well format to increase sample throughput is presented.
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References
Verslues, P.E., Agarwal, M., Katiyar-Agarwal, S., Zhu, J.H., and Zhu, J.K. (2006) Methods and concepts in quantifying resistance to drought, salt and freezing, abiotic stresses that affect plant water status. Plant J 45, 523–539.
Verslues, P.E. and Bray, E.A. (2004) LWR1 and LWR2 are required for osmoregulation and osmotic adjustment in Arabidopsis. Plant Physiol 136, 2831–2842.
Zhang, J., Nguyen, H.T., and Blum, A. (1999) Genetic analysis of osmotic adjustment in crop plants. J Exp Bot 50, 292–302.
Wood, J.M. (1999) Osmosensing by bacteria: signals and membrane-based sensors. Microbiol Mol Biol Rev 63, 230–262.
Leigh, R.A., Ahmad, N., and Wyn Jones, R.G. (1981) Assessment of glycine betaine and proline compartmentation by analysis of isolated beet vacuoles. Planta 153, 34–41.
Voetberg, G.S. and Sharp, R.E. (1991) Growth of the maize primary root at low water potentials. III. Role of increased proline deposition in osmotic adjustment. Plant Physiol 96, 1125–1130.
Hare, P.D., Cress, W.A., and van Staden, J. (1999) Proline synthesis and degradation: a model system for elucidating stress-related signal transduction. J Exp Bot 50, 413–434.
Lilley, J.M., Ludlow, M.M., McCouch, S.R., and Otoole, J.C. (1996) Locating QTL for osmotic adjustment and dehydration tolerance in rice. J Exp Bot 47, 1427–1436.
Robin, S., Pathan, M.S., Courtois, B., Lafitte, R., Carandang, S., Lanceras, S., Amante, M., Nguyen, H.T., and Li, Z. (2003) Mapping osmotic adjustment in an advanced back-cross inbred population of rice. Theor Appl Genet 107, 1288–1296.
Teulat, B., This, D., Khairallah, M., Borries, C., Ragot, C., Sourdille, P., Leroy, P., Monneveux, P., and Charrier, A. (1998) Several QTLs involved in osmotic adjustment trait variation in barley (Hordeum vulgare L.). Theor Appl Genet 96, 688–698.
Taiz, L. and Zeiger, E. (1991) Plant Physiology. Redwood City, CA: Benjamin Cummings Publishing Company.
Bates, L.S., Waldren, R.P., and Teare, I.D. (1973) Rapid determination of free proline in water-stress studies. Plant Soil 39, 205–207.
van der Weele, C.M., Spollen, W.G., Sharp, R.E., and Baskin, T.I. (2000) Growth of Arabidopsis thaliana seedlings under water deficit studied by control of water potential in nutrient-agar media. J Exp Bot 51, 1555–1562.
Acknowledgments
I thank Dr. Sandeep Sharma and Nagaraj Kumar for comments on the procedures and Shu-ya Chiang and Wei-ju Chen for assistance in the laboratory. Work in the Verslues laboratory is supported by Academia Sinica and the National Science Council of Taiwan.
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Verslues, P.E. (2010). Quantification of Water Stress-Induced Osmotic Adjustment and Proline Accumulation for Arabidopsis thaliana Molecular Genetic Studies. In: Sunkar, R. (eds) Plant Stress Tolerance. Methods in Molecular Biology, vol 639. Humana Press. https://doi.org/10.1007/978-1-60761-702-0_19
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DOI: https://doi.org/10.1007/978-1-60761-702-0_19
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