Abstract

Camellia species, an important economic plants widely distributed in Asia, are recalcitrant to RNA extraction. Here, we developed a method for high quality RNA isolation. Based on the RNA isolated from flower buds, deep transcriptome sequencing of Camellia oleifera, Camellia chekiangoleosa and Camellia brevistylawere successfully carried out. About 600,000 readings produced in a single 454 sequencing run were assembled into 49,909 contigs and 72,877 singlets, 41.06% of which were annotated. Subsequently, chalcone synthase (Chs) gene of C. chekiangoleosa was first cloned for gene expression analysis. All these works could lay the foundation for future molecular studies of Camellias.

Key words: Camellias, RNA extraction, transcriptome sequencing, gene expression analysis

Abbreviation

 CTAB, Cetyltrimethyammonium bromide; DEPC, diethyl pyrocarbonate; PVP, polyvinylpyrrolidone; RACE, rapid amplification of cDNA ends;RT-PCR, reverse transcription-polymerase chain reaction.