Transcript profiling of isolated female gametes, zygotes, and two-celled embryos
Transcriptomics, also referred to as expression profiling, is the investigation of a set (or all) RNA molecules expressed in a certain tissue or cell type, in a specific developmental stage, or under certain physiological conditions. Besides expression profiling, transcriptomics serves as a screening tool for candidate gene discovery.
We isolated female gametes (egg cells and central cells) and two-celled proembryos of wheat and Arabidopsis by manual microdissection and used these cells for transcriptome analyses. We performed both single-run partial sequencing of randomly selected cDNA clones (ESTs; “expressed sequence tags”) of generated cDNA libraries (Sprunck et al., 2005) and microarray-based expression profiling.
Our expression data sets were analyzed with the aim to discover novel gamete-specific or fertilization-induced genes. We identified transcripts specifically present in the egg cell and/or the central cell, or in zygotes and two-celled proembryos.
Currently, we investigate the roles of selected candidate genes/gene families in establishing polarity, in egg cell specification,, in gamete interaction during double fertilization, or in early embryogenesis.
The main aims and research questions of this project are:
- Comparative studies on the transcriptomes of the female gametes, of zygotes, and of two-celled proembryos, as the key to understanding gamete specification and maternal-to-zygotic transition in flowering plants
- Identification of novel players involved in gamete specification, gamete interactions during double fertilization, and the establishment of polarity