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First published online March 26, 2004; 10.1104/pp.103.036293

Plant Physiology 134:1598-1613 (2004)
© 2004 American Society of Plant Biologists

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DEVELOPMENT AND HORMONE ACTION

The Effect of {alpha}-Amanitin on the Arabidopsis Seed Proteome Highlights the Distinct Roles of Stored and Neosynthesized mRNAs during Germination1

Loïc Rajjou, Karine Gallardo2, Isabelle Debeaujon, Joël Vandekerckhove, Claudette Job and Dominique Job*

Laboratoire Mixte Centre National de la Recherche Scientifique-Bayer CropScience, Bayer CropScience, F-69263 Lyon, France (L.R., K.G., C.J., D.J.); Laboratoire de Biologie des Semences, Institut National de la Recherche Agronomique, F-78026 Versailles, France (I.D.); and Flanders Interuniversity Institute for Biotechnology and Department of Biochemistry, Gent University, B-9000 Gent, Belgium (J.V.)

To investigate the role of stored and neosynthesized mRNAs in seed germination, we examined the effect of {alpha}-amanitin, a transcriptional inhibitor targeting RNA polymerase II, on the germination of nondormant Arabidopsis seeds. We used transparent testa mutants, of which seed coat is highly permeable, to better ascertain that the drug can reach the embryo during seed imbibition. Even with the most permeable mutant (tt2-1), germination (radicle protrusion) occurred in the absence of transcription, while subsequent seedling growth was blocked. In contrast, germination was abolished in the presence of the translational inhibitor cycloheximide. Taken together, the results highlight the role of stored proteins and mRNAs for germination in Arabidopsis and show that in this species the potential for germination is largely programmed during the seed maturation process. The {alpha}-amanitin-resistant germination exhibited characteristic features. First, this germination was strongly slowed down, indicating that de novo transcription normally allows the synthesis of factor(s) activating the germination rate. Second, the sensitivity of germination to gibberellic acid was reduced 15-fold, confirming the role of this phytohormone in germination. Third, de novo synthesis of enzymes involved in reserve mobilization and resumption of metabolic activity was repressed, thus accounting for the failure in seedling establishment. Fourth, germinating seeds can recapitulate at least part of the seed maturation program, being capable of using mRNAs stored during development. Thus, commitment to germination and plant growth requires transcription of genes allowing the imbibed seed to discriminate between mRNAs to be utilized in germination and those to be destroyed.

1 The PhD thesis of L.R. is supported by Bayer CropScience and the French Ministry of Industry. J.V. was supported by the Concerted Research Actions of the Flemish Community (Belgium).

2 Present address: Unité de Génétique et Ecophysiologie des Légumineuses, Institut National de la Recherche Agronomique (INRA)-Dijon, Domaine d'Epoisses, 21110 Bretenières, France.

Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.103.036293.

* Corresponding author; e-mail dominique.job{at}bayercropscience.com; fax 33–4–72–85–22–97.

Received November 17, 2003; returned for revision January 13, 2004; accepted January 20, 2004.




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