title: A de novo transcriptome and valid reference genes for quantitative real-time PCR in Colaphellus bowringi.
abstract: BACKGROUND
  The cabbage beetle Colaphellus bowringi Baly is a serious insect pest of crucifers and undergoes reproductive diapause in soil. An understanding of the molecular mechanisms of diapause regulation, insecticide resistance, and other physiological processes is helpful for developing new management strategies for this beetle. However, the lack of genomic information and valid reference genes limits knowledge on the molecular bases of these physiological processes in this species.
  
  
  RESULTS
  Using Illumina sequencing, we obtained more than 57 million sequence reads derived from C. bowringi, which were assembled into 39,390 unique sequences. A Clusters of Orthologous Groups classification was obtained for 9,048 of these sequences, covering 25 categories, and 16,951 were assigned to 255 Kyoto Encyclopedia of Genes and Genomes pathways. Eleven candidate reference gene sequences from the transcriptome were then identified through reverse transcriptase polymerase chain reaction. Among these candidate genes, EF1α, ACT1, and RPL19 proved to be the most stable reference genes for different reverse transcriptase quantitative polymerase chain reaction experiments in C. bowringi. Conversely, aTUB and GAPDH were the least stable reference genes.
  
  
  CONCLUSIONS
  The abundant putative C. bowringi transcript sequences reported enrich the genomic resources of this beetle. Importantly, the larger number of gene sequences and valid reference genes provide a valuable platform for future gene expression studies, especially with regard to exploring the molecular mechanisms of different physiological processes in this species.
authors.affiliation: College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, P. R. China; Hubei Insect Resources Utilization and Sustainable Pest Management Key Laboratory, Huazhong Agricultural University, Wuhan, P. R. China.
authors.forename: Qian-Qian
authors.initials: QQ
authors.lastname: Tan
authors.affiliation: College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, P. R. China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhu
authors.affiliation: College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, P. R. China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Li
authors.affiliation: College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, P. R. China.
authors.forename: Wen
authors.initials: W
authors.lastname: Liu
authors.affiliation: Hubei Insect Resources Utilization and Sustainable Pest Management Key Laboratory, Huazhong Agricultural University, Wuhan, P. R. China.
authors.forename: Wei-Hua
authors.initials: WH
authors.lastname: Ma
authors.affiliation: Hubei Insect Resources Utilization and Sustainable Pest Management Key Laboratory, Huazhong Agricultural University, Wuhan, P. R. China.
authors.forename: Chao-Liang
authors.initials: CL
authors.lastname: Lei
authors.affiliation: College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, P. R. China.
authors.forename: Xiao-Ping
authors.initials: XP
authors.lastname: Wang
mesh_terms: D000818:Animals; D001517:Coleoptera / Q000196:embryology / Q000235:genetics*; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods*; D049750:Genome, Insect; D019476:Insect Proteins / Q000235:genetics / Q000592:standards*; D008297:Male; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012015:Reference Standards; D017422:Sequence Analysis, DNA
chemical_list: D019476:Insect Proteins