Thioredoxin-linked proteins are reduced during germination of Medicago truncatula seeds.
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.
Alkhalfioui
Fatima
F
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, Angers Cedex 01, France.
Renard
Michelle
M
Vensel
William H
WH
Wong
Joshua
J
Tanaka
Charlene K
CK
Hurkman
William J
WJ
Buchanan
Bob B
BB
Montrichard
Françoise
F
D000222:Adaptation, Physiological; D000255:Adenosine Triphosphate / Q000378:metabolism; D000596:Amino Acids / Q000096:biosynthesis; D001643:Bridged Bicyclo Compounds; D002244:Carbon / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism; D002473:Cell Wall / Q000378:metabolism; D018548:Cotyledon / Q000378:metabolism; D004220:Disulfides / Q000378:metabolism; D018525:Germination / Q000502:physiology*; D046913:Medicago truncatula / Q000254:growth & development / Q000378:metabolism*; D010084:Oxidation-Reduction; D010940:Plant Proteins / Q000096:biosynthesis / Q000378:metabolism*; D020543:Proteome; D040901:Proteomics; D012639:Seeds / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D013879:Thioredoxins / Q000378:metabolism*; D014815:Vitamins / Q000096:biosynthesis
D000596:Amino Acids; D001643:Bridged Bicyclo Compounds; D002352:Carrier Proteins; D004220:Disulfides; D010940:Plant Proteins; D020543:Proteome; D014815:Vitamins; D013879:Thioredoxins; D002244:Carbon; D000255:Adenosine Triphosphate; C028381:monobromobimane