PLANT PHYSIOLOGY · 2007 · 112 citations · 60 references
GeneticsCell CultureReproductive BiologyEmbryologyEmbryo CultureGerm Cell DevelopmentPublic HealthStem Cell ModelsCell DivisionMorphogenesisEmbryonic DevelopmentOrganogenesisGene ExpressionCell BiologyFunctional GenomicsMarkers AssociatedBiologyCell LineageDevelopmental BiologyAbstract Microspore-derived EmbryoCell TotipotencyEmbryo Cell IdentitySeed StorageTissue CultureMedicineCell Development
Microspore-derived embryo cultures are a model system for studying plant cell totipotency and in vitro embryo development. The study compared transcriptomic and proteomic profiles of rapeseed MDEs across developmental stages and two culture systems, aiming to identify early embryo-expressed genes and proteins for future marker development and functional studies. Global and specific expression profiles were generated using an MDE cDNA array, two-dimensional gel electrophoresis, and protein sequencing for the two culture types. Suspensor-bearing MDEs displayed enhanced histodifferentiation, revealing a potential suspensor role in cell identity and patterning, and the analysis uncovered 135 robust markers and key pathways—protein synthesis, glycolysis, and ascorbate metabolism—highlighting differential expression that may drive MDE 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.
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