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First published online March 23, 2007; 10.1104/pp.107.098723

Plant Physiology 144:155-172 (2007)
© 2007 American Society of Plant Biologists

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DEVELOPMENT AND HORMONE ACTION

Combined Transcriptome and Proteome Analysis Identifies Pathways and Markers Associated with the Establishment of Rapeseed Microspore-Derived Embryo Development1,[W]

Ronny Joosen, Jan Cordewener, Ence Darmo Jaya Supena, Oscar Vorst, Michiel Lammers, Chris Maliepaard, Tieme Zeilmaker, Brian Miki, Twan America, Jan Custers and Kim Boutilier*

Business Units Bioscience (R.J., J. Cordewener, E.D.J.S., O.V., M.L., T.Z., T.A., J. Custers, K.B.) and Biometry (C.M.), Plant Research International, 6700 AA Wageningen, The Netherlands; Research Center for Biotechnology, Bogor Agricultural University, Bogor 16610, Indonesia (E.D.J.S.); and Eastern Cereal and Oilseeds Research Centre, Agriculture and Agri-Food Canada, Ottawa, Ontario, Canada K1A 0C6 (B.M.)

Microspore-derived embryo (MDE) cultures are used as a model system to study plant cell totipotency and as an in vitro system to study embryo development. We characterized and compared the transcriptome and proteome of rapeseed (Brassica napus) MDEs from the few-celled stage to the globular/heart stage using two MDE culture systems: conventional cultures in which MDEs initially develop as unorganized clusters that usually lack a suspensor, and a novel suspensor-bearing embryo culture system in which the embryo proper originates from the distal cell of a suspensor-like structure and undergoes the same ordered cell divisions as the zygotic embryo. Improved histodifferentiation of suspensor-bearing MDEs suggests a new role for the suspensor in driving embryo cell identity and patterning. An MDE culture cDNA array and two-dimensional gel electrophoresis and protein sequencing were used to compile global and specific expression profiles for the two types of MDE cultures. Analysis of the identities of 220 candidate embryo markers, as well as the identities of 32 sequenced embryo up-regulated protein spots, indicate general roles for protein synthesis, glycolysis, and ascorbate metabolism in the establishment of MDE development. A collection of 135 robust markers for the transition to MDE development was identified, a number of which may be coregulated at the gene and protein expression level. Comparison of the expression profiles of preglobular-stage conventional MDEs and suspensor-bearing MDEs identified genes whose differential expression may reflect improved histodifferentiation of suspensor-bearing embryos. This collection of early embryo-expressed genes and proteins serves as a starting point for future marker development and gene function studies aimed at understanding the molecular regulation of cell totipotency and early embryo development in plants.

1 This work was supported by the Dutch Ministry of Agriculture, Nature, and Food Quality (program no. DWK 281–392), a Natural Science and Engineering Research Council visiting fellowship in a Canadian government laboratory, and the Biotechnology Research Indonesia-Netherlands research program, with financial aid from the Royal Netherlands Academy of Arts and Sciences and the fellowship program Quality for Undergraduate Education, Bogor Agricultural University, Indonesia.

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: Kim Boutilier (kim.boutilier{at}wur.nl).

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

www.plantphysiol.org/cgi/doi/10.1104/pp.107.098723

* Corresponding author; e-mail kim.boutilier{at}wur.nl; fax 31–317–423110.

Received March 2, 2007; accepted March 13, 2007; published March 23, 2007.




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