title: De Novo characterization of a Cephalotaxus hainanensis transcriptome and genes related to paclitaxel biosynthesis.
abstract: Cephalotaxus hainanensis, an endangered plant, is known to contain several metabolites with anti-cancer activity. Despite its clinical impact, the alkaloid metabolism of this species has remained largely uncharacterized. The potential of Cephalotaxus for metabolic engineering of medically interesting compounds has, so far, not been exploited, due to the almost complete lack of molecular information. We have therefore performed a high throughput RNA-seq analysis and assembled the transcriptome de novo. Raw reads comprising 4.3 Gbp were assembled de novo into 39,416 unique sequences (unigenes) with a mean length of 1,089.8 bp and a total assembly size of 45.8 Mbp, which equals to more than 50 times the number of Cephalotaxaceae sequences currently deposited in the GenBank (as of August 2013). As proof of principle for medically interesting pathways, gene fragments related to paclitaxel biosynthesis were searched and detected. To verify their functionality, the metabolic product paclitaxel, and its precursor baccatin III, were identified in the leaves of C. hainanensis by HPLC, and shown to be induced by MeJA. This finding demonstrates exemplarily the potential of the annotated transcriptome as information resource for the biotechnological exploitation of plant secondary metabolism.
authors.affiliation: Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China, Ministry of Agriculture/Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Qiao
authors.affiliation: Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China, Ministry of Agriculture/Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan, China.
authors.forename: Hanqing
authors.initials: H
authors.lastname: Cong
authors.affiliation: Key Laboratory of Protection and Development Utilization of Tropical Crop Germplasm Resources (Hainan University), Ministry of Education/College of Horticulture and Landscape Architecture, Hainan University, Haikou, Hainan, China.
authors.forename: Xuefei
authors.initials: X
authors.lastname: Jiang
authors.affiliation: Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China, Ministry of Agriculture/Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan, China.
authors.forename: Rongxiang
authors.initials: R
authors.lastname: Wang
authors.affiliation: Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China, Ministry of Agriculture/Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan, China.
authors.forename: Junmei
authors.initials: J
authors.lastname: Yin
authors.affiliation: Key Laboratory of Protection and Development Utilization of Tropical Crop Germplasm Resources (Hainan University), Ministry of Education/College of Horticulture and Landscape Architecture, Hainan University, Haikou, Hainan, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Qian
authors.affiliation: Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China, Ministry of Agriculture/Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan, China.
authors.forename: Zhunian
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Molecular Cell Biology, Botanical Institute, Karlsruhe Institute of Technology, Karlsruhe, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Nick
mesh_terms: D032483:Cephalotaxus / Q000235:genetics*; D018506:Gene Expression Regulation, Plant / Q000235:genetics; D017343:Genes, Plant / Q000235:genetics; D017239:Paclitaxel / Q000096:biosynthesis*; D018515:Plant Leaves / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods; D059467:Transcriptome / Q000235:genetics*
chemical_list: D017239:Paclitaxel