Generation of Expressed Sequence Tags and development of microsatellite markers for Castanopsis sieboldii var. sieboldii (Fagaceae)

• Castanopsis sieboldii var. sieboldii is an evergreen broadleaved canopy tree that grows on Honshu, Shikoku and Kyushu Islands in Japan. A closely-related, hybridizing, congener species, C. cuspidata var. cuspidata , has different leaf epidermis structure and different seed nuts morphology compared to C. sieboldii var. sieboldii . Furthermore, the habitats in which the two species grow often indicate different water requirements. • Analysis of genetic diversity, using transcribed sequences, will provide a sound basis for the management of this species, allowing successful reforestation, incorporating phylogeographic conservation. In order to obtain transcripts sequences for the species and to develop transcript-based markers, we constructed and analyzed a cDNA library, surveyed microsatellite sequences within it and designed PCR primers for the microsatellites. • The cDNA library was constructed using tissue from the inner bark of C. sieboldii var. sieboldii ; 3 354 Expressed Sequence Tags (ESTs) were identified. We constructed 2 417 putative unigenes and assigned putative functions to 1 856 of them. Three hundred and fourteen microsatellites were found within the putative unigenes and 16 EST-SSR (Simple Sequence Repeat) markers were developed. The EST-SSR markers developpe in this study should facilitate future analysis of the genetic diversity of this and related species.