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Tobacco Mesophyll Protoplasts Synthesize 1,3-beta-Glucanase, Chitinases, and "Osmotins" during in Vitro Culture
Authors:Grosset J  Meyer Y  Chartier Y  Kauffmann S  Legrand M  Fritig B
Institution:Université de Perpignan, Laboratoire de Physiologie Végétale, Avenue de Villeneuve, 66025 Perpignan Cédex, France.
Abstract:Tobacco (Nicotiana tabacum) mesophyll protoplasts synthesize six basic proteins (a, a′, a1, b, b′, and c) which are undetectable in the leaf and whose synthesis is reduced by auxin (Y Meyer, L Aspart, Y Chartier 1984] Plant Physiol 75: 1027-1033). Polypeptides a, a′, and a1 were shown to have similar mobilities on two-dimensional electrophoresis as one 1,3-β-glucanase and two chitinases from tobacco mosaic virus-infected leaves. In immunoblotting experiments, polypeptide a was recognized by specific antibodies raised against the 1,3-β-glucanase and a′ and a1 reacted with anti-chitinase antibodies. Similarly, b and b′ comigrated with osmotin and its neutral counterpart, two proteins characteristic of salt-adapted tobacco cells, and reacted with anti-osmotin antibodies. In addition it has been shown that 1,3-β-glucanase and chitinase activities increased at the same time as a, a′, and a1 accumulated in cultivated protoplasts. Finally, polypeptide c was also detected in tobacco mosaic virus-infected leaves but could not be identified as any of the pathogenesis-related proteins characterized so far in tobacco. Thus, cultivated tobacco protoplasts synthesize and accumulate typical stress proteins.
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