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Published on October 15, 2008; 10.1104/pp.108.128108

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Received August 19, 2008
Accepted October 6, 2008

Transcriptome analysis of proliferating arabidopsis endosperm reveals biological implications for the control of syncytial division, cytokinin signalling, and gene expression regulation

Robert C Day , Rowan P Herridge , Barbara A Ambrose , and Richard C Macknight *

Department of Biochemistry, University of Otago, PO Box 56, Dunedin, NZ; Institute of Molecular BioSciences, Massey University, Private Bag 11222, Palmerston North, NZ

* Corresponding author; email: richard.macknight{at}otago.ac.nz.

During the early stages of seed development, Arabidopsis thaliana (arabidopsis) endosperm is syncytial and proliferates rapidly through repeated rounds of mitosis without cytokinesis. This stage of endosperm development is important in determining final seed size and is a model for studying aspects of cellular and molecular biology, such as the cell cycle and genomic imprinting. However, the small size of arabidopsis seed makes high-throughput molecular analysis of the early endosperm technically difficult. Laser capture microdissection enabled high-resolution transcript analysis of the syncytial stage of arabidopsis endosperm development four days after pollination. Analysis of gene ontology representation revealed a developmental programme dominated by the expression of genes associated with cell cycle, DNA processing, chromatin assembly, protein synthesis, cytoskeletal and microtubule related processes, and cell/organelle biogenesis and organisation. Analysis of core cell-cycle genes implicates particular gene-family members as playing important roles in controlling syncytial cell division. Hormone marker analysis indicates predominance for cytokinin signalling during early endosperm development. Comparisons with publicly available microarray data revealed that ~800 putative early seed-specific genes were preferentially expressed in the endosperm. Early seed expression was confirmed for 71 genes using quantitative RT-PCR with 27 transcription factors being confirmed as early seed specific. Promoter-reporter lines confirmed endosperm-preferred expression at four days after pollination for five transcription factors, which validates the approach and suggests important roles for these genes during early endosperm development. In summary, the data generated provides a useful resource providing novel insight into early seed development and identifies new target genes for further characterisation.




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