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Dendrobiumofficinale( Orchidaceae ) is one of the world’s most endangered plants with great medicinal value. In nature,D.  officinaleseeds must establish symbiotic relationships with fungi to germinate. However, the molecular events involved in the interaction between fungus and plant during this process are poorly understood. To isolate the genes involved in symbiotic germination, a suppression subtractive hybridization (SSH) cDNA library of symbiotically germinatedD.  officinaleseeds was constructed. From this library, 1437 expressed sequence tags (ESTs) were clustered to 1074  Unigenes  (including 902 singletons and 172 contigs), which were searched against the NCBI non-redundant (NR) protein database (E-value cutoff, e -5 ). Based on sequence similarity with known proteins, 579 differentially expressed genes inD.  officinalewere identified and classified into different functional categories by Gene Ontology (GO), Clusters of orthologous Groups of proteins (COGs) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. The expression levels of 15 selected genes emblematic of symbiotic germination were confirmed via real-time quantitative PCR. These genes were classified into various categories, including defense and stress response, metabolism, transcriptional regulation, transport process and signal transduction pathways. All transcripts were upregulated in the symbiotically germinated seeds (SGS). The functions of these genes in symbiotic germination were predicted. Furthermore, two fungus-induced calcium-dependent protein kinases (CDPKs), which were upregulated 6.76- and 26.69-fold in SGS compared with un-germinated seeds (UGS), were cloned fromD.  officinaleand characterized for the first time. This study provides the first global overview of genes putatively involved inD.  officinalesymbiotic seed germination and provides a foundation for further functional research regarding symbiotic relationships in orchids.

ESTs Analysis Reveals Putative Genes Involved in Symbiotic Seed Germination in Dendrobium officinale