title: Mining and identification of polyunsaturated fatty acid synthesis genes active during camelina seed development using 454 pyrosequencing.
abstract: BACKGROUND
  Camelina (Camelina sativa L.) is well known for its high unsaturated fatty acid content and great resistance to environmental stress. However, little is known about the molecular mechanisms of unsaturated fatty acid biosynthesis in this annual oilseed crop. To gain greater insight into this mechanism, the transcriptome profiles of seeds at different developmental stages were analyzed by 454 pyrosequencing.
  
  
  RESULTS
  Sequencing of two normalized 454 libraries produced 831,632 clean reads. A total of 32,759 unigenes with an average length of 642 bp were obtained by de novo assembly, and 12,476 up-regulated and 12,390 down-regulated unigenes were identified in the 20 DAF (days after flowering) library compared with the 10 DAF library. Functional annotations showed that 220 genes annotated as fatty acid biosynthesis genes were up-regulated in 20 DAF sample. Among them, 47 candidate unigenes were characterized as responsible for polyunsaturated fatty acid synthesis. To verify unigene expression levels calculated from the transcriptome analysis results, quantitative real-time PCR was performed on 11 randomly selected genes from the 220 up-regulated genes; 10 showed consistency between qRT-PCR and 454 pyrosequencing results.
  
  
  CONCLUSIONS
  Investigation of gene expression levels revealed 32,759 genes involved in seed development, many of which showed significant changes in the 20 DAF sample compared with the 10 DAF sample. Our 454 pyrosequencing data for the camelina transcriptome provide an insight into the molecular mechanisms and regulatory pathways of polyunsaturated fatty acid biosynthesis in camelina. The genes characterized in our research will provide candidate genes for the genetic modification of crops.
authors.affiliation: Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, Jilin, 130118, China. fw-1980@163.com.
authors.forename: Fawei
authors.initials: F
authors.lastname: Wang
authors.affiliation: College of life Sciences, Jilin Agricultural University, Changchun, Jilin, 130118, China. chjlau@163.com.
authors.forename: Huan
authors.initials: H
authors.lastname: Chen
authors.affiliation: Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, Jilin, 130118, China. xiaoweili1206@163.com.
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Li
authors.affiliation: Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, Jilin, 130118, China. wangnanlunwen@126.com.
authors.forename: Nan
authors.initials: N
authors.lastname: Wang
authors.affiliation: College of life Sciences, Jilin Agricultural University, Changchun, Jilin, 130118, China. 285064134@qq.com.
authors.forename: Tianyi
authors.initials: T
authors.lastname: Wang
authors.affiliation: Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, Jilin, 130118, China. yangjing5122010@163.com.
authors.forename: Jing
authors.initials: J
authors.lastname: Yang
authors.affiliation: Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, Jilin, 130118, China. guanll2004@163.com.
authors.forename: Lili
authors.initials: L
authors.lastname: Guan
authors.affiliation: Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, Jilin, 130118, China. 52730603@qq.com.
authors.forename: Na
authors.initials: N
authors.lastname: Yao
authors.affiliation: Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, Jilin, 130118, China. dulinna0918@163.com.
authors.forename: Linna
authors.initials: L
authors.lastname: Du
authors.affiliation: Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, Jilin, 130118, China. nifengcao_2000@163.com.
authors.forename: Yanfang
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, Jilin, 130118, China. xiuming1211@163.com.
authors.forename: Xiuming
authors.initials: X
authors.lastname: Liu
authors.affiliation: Jilin Technology Innovation Center for Soybean Region, Jilin Agricultural University, Changchun, Jilin, 130118, China. chenxifeng2003@126.com.
authors.forename: Xifeng
authors.initials: X
authors.lastname: Chen
authors.affiliation: Jilin Technology Innovation Center for Soybean Region, Jilin Agricultural University, Changchun, Jilin, 130118, China. wangzhenmin2004@sina.com.
authors.forename: Zhenmin
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, Jilin, 130118, China. dongyuanyuan_dyy@yahoo.com.cn.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Dong
authors.affiliation: Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, Jilin, 130118, China. hyli99@163.com.|College of life Sciences, Jilin Agricultural University, Changchun, Jilin, 130118, China. hyli99@163.com.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Li
mesh_terms: D053898:Biosynthetic Pathways / Q000235:genetics; D019607:Brassicaceae / Q000235:genetics* / Q000254:growth & development*; D057225:Data Mining; D005231:Fatty Acids, Unsaturated / Q000096:biosynthesis*; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D006358:Hot Temperature; D058977:Molecular Sequence Annotation; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D012639:Seeds / Q000235:genetics* / Q000254:growth & development*; D017422:Sequence Analysis, DNA / Q000379:methods*; D059467:Transcriptome / Q000235:genetics
chemical_list: D005231:Fatty Acids, Unsaturated