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First published online May 18, 2007; 10.1104/pp.107.098103 Plant Physiology 144:1559-1579 (2007) © 2007 American Society of Plant Biologists OPEN ACCESS ARTICLE
Thioredoxin-Linked Proteins Are Reduced during Germination of Medicago truncatula Seeds1,[W],[OA]Physiologie Moléculaire des Semences, Unité Mixte de Recherche 1191, Université d'Angers, Institut National d'Horticulture, Institut National de la Recherche Agronomique, Anjou Recherche Semences, 49045 Angers cedex 01, France (F.A., M.R., F.M.); Western Regional Research Center, United States Department of Agriculture, Agricultural Research Service, Albany, California 94710 (W.H.V., C.K.T., W.J.H.); and Department of Plant and Microbial Biology, University of California, Berkeley, California 947203102 (J.W., B.B.B.)
Germination of cereals is accompanied by extensive change in the redox state of seed proteins. Proteins present in oxidized form in dry seeds are converted to the reduced state following imbibition. Thioredoxin (Trx) appears to play a role in this transition in cereals. It is not known, however, whether Trx-linked redox changes are restricted to cereals or whether they take place more broadly in germinating seeds. To gain information on this point, we have investigated a model legume, Medicago truncatula. Two complementary gel-based proteomic approaches were followed to identify Trx targets in seeds: Proteins were (1) labeled with a thiol-specific probe, monobromobimane (mBBr), following in vitro reduction by an NADP/Trx system, or (2) isolated on a mutant Trx affinity column. Altogether, 111 Trx-linked proteins were identified with few differences between axes and cotyledons. Fifty nine were new, 34 found previously in cereal or peanut seeds, and 18 in other plants or photosynthetic organisms. In parallel, the redox state of proteins assessed in germinating seeds using mBBr revealed that a substantial number of proteins that are oxidized or partly reduced in dry seeds became more reduced upon germination. The patterns were similar for proteins reduced in vivo during germination or in vitro by Trx. In contrast, glutathione and glutaredoxin were less effective as reductants in vitro. Overall, more than half of the potential targets identified with the mBBr labeling procedure were reduced during germination. The results provide evidence that Trx functions in the germination of seeds of dicotyledons as well as monocotyledons.
1 This work was supported by the France-Berkeley fund, the Contrat Etat Region des Pays de la Loire 2000 to 2006, and the California Agricultural Experiment Station. F.A. was supported by a fellowship from the Conseil Général du Maine et Loire. The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Françoise Montrichard (francoise.montrichard{at}univ-angers.fr). [W] The online version of this article contains Web-only data. [OA] Open Access articles can be viewed online without a subscription. www.plantphysiol.org/cgi/doi/10.1104/pp.107.098103 * Corresponding author; e-mail francoise.montrichard{at}univ-angers.fr; fax 33241225549. Received February 15, 2007; accepted April 30, 2007; published May 18, 2007. This article has been cited by other articles:
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