Plant Physiology 138: 1538-1551 (2005)
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Water Uptake and Distribution in Germinating Tobacco Seeds Investigated in Vivo by Nuclear Magnetic Resonance Imaging [w]
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Bertram Manz, Kerstin Müller, Birgit Kucera, Frank Volke, Gerhard Leubner-Metzger
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Fraunhofer-Institut für Biomedizinische Technik (IBMT), Arbeitsgruppe Magnetische Resonanz, Ensheimer Str. 48, D-66386 St. Ingbert, Germany, Web: http://www.nmr.fraunhofer.de (B.M., F.V.)
Institut für Biologie II (Botanik/Pflanzenphysiologie), Albert-Ludwigs-Universität Freiburg,
Schänzlestr. 1, D-79104 Freiburg i. Br., Germany, Web: 'The Seed Biology Place' http://www.seedbiology.de (K.M., B.K., G.L.-M.)
Received February 21, 2005; revised April 14, 2005; accepted Aprill 20, 2005; published June 24, 2005
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Abstract. The regulation of water uptake of germinating tobacco (Nicotiana tabacum) seeds was studied spatially and temporally by in vivo 1H-nuclear magnetic resonance (NMR) microimaging and 1H-magic angle spinning NMR spectroscopy. These nondestructive state-of-the-art methods showed that water distribution in the water uptake phases II and III is inhomogeneous. The micropylar seed end is the major entry point of water. The micropylar endosperm and the radicle show the highest hydration. Germination of tobacco follows a distinct pattern of events: rupture of the testa is followed by rupture of the endosperm. Abscisic acid (ABA) specifically inhibits endosperm rupture and phase III water uptake, but does not alter the spatial and temporal pattern of phase I and II water uptake. Testa rupture was associated with an increase in water uptake due to initial embryo elongation, which was not inhibited by ABA. Overexpression of ß-1,3-glucanase in the seed-covering layers of transgenic tobacco seeds did not alter the moisture sorption isotherms or the spatial pattern of water uptake during imbibition, but partially reverted the ABA inhibition of phase III water uptake and of endosperm rupture. In vivo 13C-magic angle spinning NMR spectroscopy showed that seed oil mobilization is not inhibited by ABA. ABA therefore does not inhibit germination by preventing oil mobilization or by decreasing the water-holding capacity of the micropylar endosperm and the radicle. Our results support the proposal that different seed tissues and organs hydrate at different extents and that the micropylar endosperm region of tobacco acts as a water reservoir for the embryo.
Key words: abscisic acid, endosperm rupture, imbibition, ß-1,3-glucanase, NMR microimaging, oil mobilisation, seed germination, testa rupture, water uptake
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