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GENETICS, GENOMICS, AND MOLECULAR EVOLUTION

The Proteome of Seed Development in the Model Legume Lotus japonicus^1,[C],[W]

Centre for Carbohydrate Recognition and Signalling (S.D., B.S.L., J.H.Ø., B.J., L.K., J.S.), Center for Insoluble Protein Structures (I.B.T., J.J.E.), and Department of Molecular Biology (S.D., B.S.L., J.H.Ø., B.J., L.K., I.B.T., J.J.E., J.S.), University of Aarhus, DK–8000 Aarhus, Denmark; Center for Biological Sequence Analysis, Technical University of Denmark, DK–2800 Kgs Lyngby, Denmark (H.H.S., C.F., K.N.); Institut Louis Malardé, 98713 Papeete Tahiti, French Polynesia (N.G.); Australian Research Council Centre of Excellence for Integrative Legume Research, Genomic Interactions Group, Research School of Biological Sciences, Australian National University, Canberra, Australian Capital Territory 2601, Australia (N.G.); Bioinformatics Research Center, University of Aarhus, DK–8000 Aarhus, Denmark (S.B.); and Kazusa DNA Research Institute, Kisarazu, Chiba 292–0818, Japan (S.S., S.T.)

We have characterized the development of seeds in the model legume Lotus japonicus. Like soybean (Glycine max) and pea (Pisum sativum), Lotus develops straight seed pods and each pod contains approximately 20 seeds that reach maturity within 40 days. Histological sections show the characteristic three developmental phases of legume seeds and the presence of embryo, endosperm, and seed coat in desiccated seeds. Furthermore, protein, oil, starch, phytic acid, and ash contents were determined, and this indicates that the composition of mature Lotus seed is more similar to soybean than to pea. In a first attempt to determine the seed proteome, both a two-dimensional polyacrylamide gel electrophoresis approach and a gel-based liquid chromatography-mass spectrometry approach were used. Globulins were analyzed by two-dimensional polyacrylamide gel electrophoresis, and five legumins, LLP1 to LLP5, and two convicilins, LCP1 and LCP2, were identified by matrix-assisted laser desorption ionization quadrupole/time-of-flight mass spectrometry. For two distinct developmental phases, seed filling and desiccation, a gel-based liquid chromatography-mass spectrometry approach was used, and 665 and 181 unique proteins corresponding to gene accession numbers were identified for the two phases, respectively. All of the proteome data, including the experimental data and mass spectrometry spectra peaks, were collected in a database that is available to the scientific community via a Web interface (http://www.cbs.dtu.dk/cgi-bin/lotus/db.cgi). This database establishes the basis for relating physiology, biochemistry, and regulation of seed development in Lotus. Together with a new Web interface (http://bioinfoserver.rsbs.anu.edu.au/utils/PathExpress4legumes/) collecting all protein identifications for Lotus, Medicago, and soybean seed proteomes, this database is a valuable resource for comparative seed proteomics and pathway analysis within and beyond the legume family.

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: Jens Stougaard (stougaard{at}mb.au.dk).

^[C] Some figures in this article are displayed in color online but in black and white in the print edition.

^[W] The online version of this article contains Web-only data.

www.plantphysiol.org/cgi/doi/10.1104/pp.108.133405

^* Corresponding author; e-mail stougaard{at}mb.au.dk.

Received November 28, 2008; accepted January 3, 2009; published January 7, 2009.

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