Sequencing technology has been rapidly advancing. Giga-sequencers, which produce several gigabases of fragmented sequences per run, are attractive for decoding genomes and expressed sequence tags (ESTs). A variety of plant genomes and ESTs have been sequenced since the decoding of the genome of Arabidopsis thaliana, the model plant. ESTs are useful for functional analyses of genes and proteins and as biomarkers, which are used to identify particular tissues and conditions due to the specificity of their expression. Sequenced plant genomes and ESTs have been entered into public databases, where they are freely downloadable. Sequences representative of particular functions or structures have been collected from public databases to curate smaller databases useful for studying protein function. Here, we discuss the uses of the currently available plant EST datasets. We also demonstrate the use of network module analysis to perform more stable (or irrespective of the difference of performance in each analyzing PC) homology searches and to provide more information on molecular functions of plant ESTs and proteins.Key words: Database, expressed sequence tag (EST), homology search, network module analysis.Plant Biotechnology 28, 351-360 (2011) DOI: 10.5511/plantbiotechnology.11.0818a Abbreviations: BLAST, Basic Local Alignment Search Tool; EST, expressed sequence tags; GBFF, GenBank flatfile; GFF, General Feature Format; GPFF, GenPept flatfime. This article can be found at http://www.jspcmb.jp/ Published online September 25, 2011 perform more stable homology searches and to provide more information on molecular functions of ESTs, downsizing while maintaining the precision of homology search and also constructing local modules, in which sequences are highly homologous and thus belong to a group with a common feature, are useful. The PSI-BLAST algorithm and its derivatives (Altschul et al. 1997;Lee et al. 2009;Li et al. 2011) focus on sequenceto-sequence hits between multiple sequences. To evaluate relationships between multiple elements (e.g., gene or metabolite), network module analysis is a useful approach (Saito et al. 2008). Network module analyses have been applied to plant gene co-expression, in which a plant gene is related to other genes based on similar expression profiles (Aoki et al. 2007;Ficklin and Feltus 2011;Huber et al. 2007;Marino-Ramirez et al. 2009;Ogata et al. 2010;Winden et al. 2011). This approach allows a co-expression module, which includes coexpressed gene to be assigned to a particular biological process. By identifying homologies between sequences, network module analysis can be used to create, a homology network in which a sequence (node) is connected to other sequences on the basis of high homology. To perform such analysis for a homology network, we used our algorithm (Ogata et al. 2009) according to the following processes: 1) performing BLAST for any pairs of sequences, 2) calculating association indices between pairs as described in "Userfriendly tools for using plant EST a...