Laser capture microdissection for the analysis of gene expression during embryogenesis of Arabidopsis.
It is during embryogenesis that the body plan of the developing plant is established. Analysis of gene expression during embryogenesis has been limited due to the technical difficulty of accessing the developing embryo. Here we demonstrate that laser capture microdissection can be applied to the analysis of embryogenesis. We show how this technique can be used in concert with DNA microarray for the large-scale analysis of gene expression in apical and basal domains of the globular-stage and heart-stage embryo, respectively, when critical events of polarity, symmetry and biochemical differentiation are established. This high resolution spatial analysis shows that up to approximately 65% of the genome is expressed in the developing embryo, and that differential expression of a number of gene classes can be detected. We discuss the validity of this approach for the functional analysis of both published and previously uncharacterized essential genes.
Casson
Stuart
S
The Integrative Cell Biology Laboratory, School of Biological and Biomedical Sciences, University of Durham, South Road, Durham DH1 3LE, UK.
Spencer
Matthew
M
Walker
Katherine
K
Lindsey
Keith
K
D017360:Arabidopsis / Q000196:embryology* / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D018506:Gene Expression Regulation, Plant / Q000502:physiology*; D042282:Microdissection / Q000379:methods*; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype