By Sabine Globig; Philip Stewart

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Automated Sequence Assemblies of PGB02 and PGB04 The shotgun plasmid libraries for PGB02 and PGB04 were arrayed in 384-well plates. Plasmid inserts from 10 and five 384-well plates were Sanger-sequenced for PGB02 and PGB04, respectively, resulting in 6,954 and 3,677 paired sequence reads. The average plasmid insert length was 1,102 bp for the PGB02 library and 1,056 bp for the PGB04 library. 9% (PGB04) of the total sequences. Using PHRAP, we assembled the sequences into 15 contigs for PGB02 and 14 contigs for PGB04.

Clustered cycad ESTs were assigned to a functional category based on top BLASTP similarity scores. An expect value (E-value) of > 1e-10 was chosen as the cut-off threshold. The analysis was performed at the Munich Information Center for Protein Sequences. Cycad Contig Matches to Genes in Angiosperms, Gymnosperms, and Lower Plants Using TBLASTX, a comparison was made between the C. rumphii UniGene set versus all available ESTs from GenBank and predicted Arabidopsis genes from The Arabidopsis Information Resource (TAIR).

The COP9 encodes a subunit of the COP9 signalosome complex, which controls multiple signaling pathways that regulate development in all eukaryotes (Chamovitz and Glickman, 2002; Hellmann and Estelle, 2002). In Arabidopsis, the COP9 mutant is constitutively photomorphogenic in dark-grown seedlings (Wei and Deng, 1992). , 1996). As yet, the phenotype of dark-grown cycad seedlings has not been fully evaluated. The discovery of a gene encoding a putative 28 Vascular Plants and Paleobotany subunit of the COP9 complex in cycads could be a first step to define the ancestral, developmental role of the signalosome in gymnosperms, particularly with regard to its role in photomorphogenesis.

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