title: Combined transcriptome and proteome analysis identifies pathways and markers associated with the establishment of rapeseed microspore-derived embryo development.
abstract: 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.
authors: Business Units Bioscience, Plant Research International, 6700 AA Wageningen, The Netherlands.
authors: Ronny
authors: R
authors: Joosen
authors: Jan
authors: J
authors: Cordewener
authors: Ence Darmo Jaya
authors: ED
authors: Supena
authors: Oscar
authors: O
authors: Vorst
authors: Michiel
authors: M
authors: Lammers
authors: Chris
authors: C
authors: Maliepaard
authors: Tieme
authors: T
authors: Zeilmaker
authors: Brian
authors: B
authors: Miki
authors: Twan
authors: T
authors: America
authors: Jan
authors: J
authors: Custers
authors: Kim
authors: K
authors: Boutilier
mesh_terms: D029688:Brassica napus / Q000196:embryology / Q000235:genetics* / Q000378:metabolism; D016000:Cluster Analysis; D047108:Embryonic Development; D020869:Gene Expression Profiling; D005819:Genetic Markers; D010940:Plant Proteins / Q000378:metabolism*; D020543:Proteome; D012333:RNA, Messenger / Q000378:metabolism*; D046509:Tissue Culture Techniques
chemical_list: D005819:Genetic Markers; D010940:Plant Proteins; D020543:Proteome; D012333:RNA, Messenger