Abstract
The anoxia-dependent elevation of cytosolic Ca2+ concentration, [Ca2+]cyt, was investigated in plants differing in tolerance to hypoxia. The [Ca2+]cyt was measured by fluorescence microscopy in single protoplasts loaded with the calcium-fluoroprobe Fura 2-AM. Imposition of anoxia led to a fast (within 3 min) significant elevation of [Ca2+]cyt in rice leaf protoplasts. A tenfold drop in the external Ca2+ concentration (to 0.1 mM) resulted in considerable decrease of the [Ca2+]cyt shift. Rice root protoplasts reacted upon anoxia with higher amplitude. Addition of plasma membrane (verapamil, La3+ and EGTA) and intracellular membrane Ca2+-channel antagonists (Li+, ruthenium red and cyclosporine A) reduced the anoxic Ca2+-accumulation in rice. Wheat protoplasts responded to anoxia by smaller changes of [Ca2+]cyt. In wheat leaf protoplasts, the amplitude of the Ca2+-shift little depended on the external level of Ca2+. Wheat root protoplasts were characterized by a small shift of [Ca2+]cyt under anoxia. Plasmalemma Ca2+-channel blockers had little effect on the elevation of cytosolic Ca2+ in wheat protoplasts. Intact rice seedlings absorbed Ca2+ from the external medium under anoxic treatment. On the contrary, wheat seedlings were characterized by leakage of Ca2+. Verapamil abolished the Ca2+ influx in rice roots and Ca2+ efflux from wheat roots. Anoxia-induced [Ca2+]cyt elevation was high particularly in rice, a hypoxia-tolerant species. In conclusion, both external and internal Ca2+ stores are important for anoxic [Ca2+]cyt elevation in rice, whereas the hypoxia-intolerant wheat does not require external sources for [Ca2+]cyt rise. Leaf and root protoplasts similarly responded to anoxia, independent of their organ origin.
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Abbreviations
- Cs A:
-
Cyclosporine A
- DMSO:
-
Dimethyl sulfoxide
- EGTA:
-
Ethylene glycol-bis(beta-aminoethyl ether) N,N,N,N-tetra acetic acid
- Fura 2-AM:
-
Acetoxymethyl ester of calcium binding benzofuran
- RR:
-
Ruthenium red
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Acknowledgments
Financial support from the Swedish Institute, Swedish Royal Agricultural Academy (KSLA), Knut and Alice Wallenberg Foundation, Russian Foundation for Basic Research (07-04-01056a and 10-04-01035a) and Russian Ministry of Education and Science (2006-RI-111.0/002/037 and 2010-1.3.2-203-002-008) is gratefully acknowledged. Authors would like to thank Dr. O.B. Blokhina (University of Helsinki) for the help with Oxygrath measurements.
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Yemelyanov, V.V., Shishova, M.F., Chirkova, T.V. et al. Anoxia-induced elevation of cytosolic Ca2+ concentration depends on different Ca2+ sources in rice and wheat protoplasts. Planta 234, 271–280 (2011). https://doi.org/10.1007/s00425-011-1396-x
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DOI: https://doi.org/10.1007/s00425-011-1396-x