Background Body color and coloration patterns are important phenotypic traits to maintain survival and reproduction activities. The Oujiang color varieties of common carp (Cyprinus carpio var. color), with a narrow distribution in Zhejiang Province of China and a history of aquaculture for over 1,200 years, consistently exhibit a variety of body color patterns. The molecular mechanism underlying diverse color patterns in these variants is unknown. To the practical end, it is essential to develop molecular markers that can distinguish different phenotypes and assist selective breeding. Methodology/Principal Findings In this exploratory study, we conducted Roche 454 transcriptome sequencing of two pooled skin tissue samples of Oujiang common carp, which correspond to distinct color patterns, red with big black spots (RB) and whole white (WW), and a total of 737,525 sequence reads were generated. The reads obtained in this study were co-assembled jointly with common carp Roche 454 sequencing reads downloaded from NCBI SRA database, resulting in 43,923 isotigs and 546,676 singletons. Over 31 thousand (31,445; 71.6%) isotigs were found with significant BLAST matches (E<1e-10) to the nr protein database, which corresponds to 12,597 annotated zebrafish genes. A total of 70,947 isotigs and singletons (transcripts) were annotated with Gene Ontology, and 60,221 transcripts were found with corresponding EC numbers. Out of 145 zebrafish pigmentation genes, orthologs for 117 were recovered in Oujiang color carp transcriptome, including 18 found only among singletons. Our transcriptome analysis revealed over 52,902 SNPs in Oujiang common carp, and identified 63 SNP markers that are putatively unique either for RB or WW. Conclusions The transcriptome of Oujiang color varieties of common carp obtained through this study, along with the pigmentation genes recovered and the color pattern-specific molecular markers developed, will facilitate future research on the molecular mechanism of color patterns and promote aquaculture of Oujiang color varieties of common carp through molecular marker assisted-selective breeding.
References
[1]
Protas ME, Patel NH (2008) Evolution of coloration patterns. Annu Rev Cell Dev Biol 24: 425–446. doi: 10.1146/annurev.cellbio.24.110707.175302
[2]
Rodgers GM, Kelley JL, Morrell LJ (2010) Colour change and assortment in the western rainbowfish. Animal Behaviour 79: 1025–1030. doi: 10.1016/j.anbehav.2010.01.017
[3]
Steiner CC, Weber JN, Hoekstra HE (2007) Adaptive variation in beach mice produced by two interacting pigmentation genes. PLoS biology 5: 1–e219. doi: 10.1371/journal.pbio.0050219
[4]
Braasch I, Schartl M, Volff JN (2007) Evolution of pigment synthesis pathways by gene and genome duplication in fish. BMC Evol Biol 7: 74. doi: 10.1186/1471-2148-7-74
[5]
Bagnara JT, Fernandez PJ, Fujii R (2007) On the blue coloration of vertebrates. Pigment Cell Res 20: 14–26. doi: 10.1111/j.1600-0749.2006.00360.x
[6]
Braasch I, Brunet F, Volff JN, Schartl M (2009) Pigmentation pathway evolution after whole-genome duplication in fish. Genome Biol Evol 1: 479–493. doi: 10.1093/gbe/evp050
[7]
Mellgren EM, Johnson SL (2002) The evolution of morphological complexity in zebrafish stripes. Trends Genet 18: 128–134. doi: 10.1016/s0168-9525(01)02614-2
[8]
Li S, Wang C, Yu W, Zhao S, Gong Y (2012) Identification of genes related to white and black plumage formation by RNA-Seq from white and black feather bulbs in ducks. PLoS One 7: e36592. doi: 10.1371/journal.pone.0036592
[9]
Logan DW, Burn SF, Jackson IJ (2006) Regulation of pigmentation in zebrafish melanophores. Pigment Cell Res 19: 206–213. doi: 10.1111/j.1600-0749.2006.00307.x
Braasch I, Liedtke D, Volff JN, Schartl M (2009) Pigmentary function and evolution of tyrp1 gene duplicates in fish. Pigment Cell Melanoma Res 22: 839–850. doi: 10.1111/j.1755-148x.2009.00614.x
[12]
Hoekstra HE, Hirschmann RJ, Bundey RA, Insel PA, Crossland JP (2006) A single amino acid mutation contributes to adaptive beach mouse color pattern. Science 313: 101–104. doi: 10.1126/science.1126121
[13]
Sugie A, Terai Y, Ota R, Okada N (2004) The evolution of genes for pigmentation in African cichlid fishes. Gene 343: 337–346. doi: 10.1016/j.gene.2004.09.019
[14]
Altschmied J, Delfgaauw J, Wilde B, Duschl J, Bouneau L, et al. (2002) Subfunctionalization of duplicate mitf genes associated with differential degeneration of alternative exons in fish. Genetics 161: 259–267.
[15]
Balon EK (1995) Origin and domestication of the wild carp,Cyprinus carpio: from Roman gourmets to the swimming flowers. Aquaculture 129: 3–48. doi: 10.1016/0044-8486(94)00227-f
[16]
Wang C, Li S, Nagy ZT, Lehoczky I, Huang L, et al. (2010) Molecular genetic structure and relationship of Chinese and Hungarian common carp (Cyprinus carpio L.) strains based on mitochondrial sequence. Aquaculture Research 41: 1339–1347. doi: 10.1111/j.1365-2109.2009.02422.x
[17]
David L, Rothbard S, Rubinstein I, Katzman H, Hulata G, et al. (2004) Aspects of red and black color inheritance in the Japanese ornamental (Koi) carp (Cyprinus carpio L.). Aquaculture 233: 129–147. doi: 10.1016/j.aquaculture.2003.10.033
[18]
Li S (2003) Aquaculture research and its relation to development in China. Agricultural development and the opportunities for aquatic resources research in China 65: 17.
[19]
Wang CH, Xiang SP, Lv YP, Wang J, Pan ZY, et al. (2008) Preliminary study on inheritance of red and white color phenotypes in Oujiang color carp, C. carpio var. color. Journal of Shanghai Fisheries University 4: 004.
[20]
David L, Blum S, Feldman MW, Lavi U, Hillel J (2003) Recent Duplication of the Common Carp (Cyprinus carpio L.) Genome as Revealed by Analyses of Microsatellite Loci. Molecular Biology and Evolution 20: 1425–1434. doi: 10.1093/molbev/msg173
[21]
Ma Y, Wang C, Wang J, Yang X, Bi X, et al. (2011) Genetic differentiation of wild and hatchery Oujiang color common carp: potential application to the identification of escapees. Fisheries Science 77: 591–597. doi: 10.1007/s12562-011-0359-2
[22]
Zhu L, Ma Y, Xiang S, Bi X, Wang J, et al. (2013) Observation and analysis of growth dynamics in Oujiang color common carp with different pigmentation types. Journal of Shanghai Fisheries University In Press.
[23]
Wang Z, Gerstein M, Snyder M (2009) RNA-Seq: a revolutionary tool for transcriptomics. Nature Reviews Genetics 10: 57–63. doi: 10.1038/nrg2484
[24]
Ji P, Liu G, Xu J, Wang X, Li J, et al. (2012) Characterization of common carp transcriptome: sequencing, de novo assembly, annotation and comparative genomics. PLoS One 7: e35152. doi: 10.1371/journal.pone.0035152
[25]
Liao X, Cheng L, Xu P, Lu G, Wachholtz M, et al. (2013) Transcriptome Analysis of Crucian Carp (Carassius auratus), an Important Aquaculture and Hypoxia-Tolerant Species. PLoS One 8: e62308. doi: 10.1371/journal.pone.0062308
[26]
Salem M, Rexroad CE 3rd, Wang J, Thorgaard GH, Yao J (2010) Characterization of the rainbow trout transcriptome using Sanger and 454-pyrosequencing approaches. BMC Genomics 11: 564. doi: 10.1186/1471-2164-11-564
[27]
Samanta MP, Tongprasit W, Istrail S, Cameron RA, Tu Q, et al. (2006) The transcriptome of the sea urchin embryo. Science 314: 960–962. doi: 10.1126/science.1131898
[28]
Shen GM, Dou W, Niu JZ, Jiang HB, Yang WJ, et al. (2011) Transcriptome analysis of the oriental fruit fly (Bactrocera dorsalis). PLoS One 6: e29127. doi: 10.1371/journal.pone.0029127
[29]
Huan P, Wang H, Liu B (2012) Transcriptomic analysis of the clam Meretrix meretrix on different larval stages. Marine Biotechnology 14: 69–78. doi: 10.1007/s10126-011-9389-0
[30]
Mu Y, Ding F, Cui P, Ao J, Hu S, et al. (2010) Transcriptome and expression profiling analysis revealed changes of multiple signaling pathways involved in immunity in the large yellow croaker during Aeromonas hydrophila infection. BMC Genomics 11: 506. doi: 10.1186/1471-2164-11-506
[31]
Visser WE, Swagemakers SM, ?zg?r Z, Schot R, Verheijen FW, et al. (2010) Transcriptional profiling of fibroblasts from patients with mutations in MCT8 and comparative analysis with the human brain transcriptome. Human molecular genetics 19: 4189–4200. doi: 10.1093/hmg/ddq337
[32]
Li C, Zhang Y, Wang R, Lu J, Nandi S, et al. (2012) RNA-seq analysis of mucosal immune responses reveals signatures of intestinal barrier disruption and pathogen entry following Edwardsiella ictaluri infection in channel catfish, Ictalurus punctatus. Fish & Shellfish Immunology 32: 816–827. doi: 10.1016/j.fsi.2012.02.004
[33]
Chouvarine P, Cooksey AM, McCarthy FM, Ray DA, Baldwin BS, et al. (2012) Transcriptome-based differentiation of closely-related Miscanthus lines. PLoS One 7: e29850. doi: 10.1371/journal.pone.0029850
[34]
Ma K, Qiu G, Feng J, Li J (2012) Transcriptome analysis of the oriental river prawn, Macrobrachium nipponense using 454 pyrosequencing for discovery of genes and markers. PLoS One 7: e39727. doi: 10.1371/journal.pone.0039727
[35]
Pereiro P, Balseiro P, Romero A, Dios S, Forn-Cuni G, et al. (2012) High-throughput sequence analysis of turbot (Scophthalmus maximus) transcriptome using 454-pyrosequencing for the discovery of antiviral immune genes. PLoS One 7: e35369. doi: 10.1371/journal.pone.0035369
[36]
Renaut S, Nolte AW, Bernatchez L (2010) Mining transcriptome sequences towards identifying adaptive single nucleotide polymorphisms in lake whitefish species pairs (Coregonus spp. Salmonidae). Mol Ecol 19 Suppl 1: 115–131. doi: 10.1111/j.1365-294x.2009.04477.x
[37]
Ji P, Zhang Y, Li C, Zhao Z, Wang J, et al. (2012) High throughput mining and characterization of microsatellites from common carp genome. Int J Mol Sci 13: 9798–9807. doi: 10.3390/ijms13089798
[38]
Ng P, Wei CL, Sung WK, Chiu KP, Lipovich L, et al. (2005) Gene identification signature (GIS) analysis for transcriptome characterization and genome annotation. Nat Methods 2: 105–111. doi: 10.1038/nmeth733
[39]
Margulies M, Egholm M, Altman WE, Attiya S, Bader JS, et al. (2005) Genome sequencing in microfabricated high-density picolitre reactors. Nature 437: 376–380.
Conesa A, Gotz S, Garcia-Gomez JM, Terol J, Talon M, et al. (2005) Blast2GO: a universal tool for annotation, visualization and analysis in functional genomics research. Bioinformatics 21: 3674–3676. doi: 10.1093/bioinformatics/bti610
[42]
Li H, Durbin R (2010) Fast and accurate long-read alignment with Burrows-Wheeler transform. Bioinformatics 26: 589–595. doi: 10.1093/bioinformatics/btp698
[43]
Xu F, Wang W, Wang P, Jun Li M, Chung Sham P, et al. (2012) A fast and accurate SNP detection algorithm for next-generation sequencing data. Nat Commun 3: 1258. doi: 10.1038/ncomms2256
[44]
Bai X, Mamidala P, Rajarapu SP, Jones SC, Mittapalli O (2011) Transcriptomics of the bed bug (Cimex lectularius). PLoS One 6: e16336. doi: 10.1371/journal.pone.0016336
[45]
Calduch-Giner JA, Bermejo-Nogales A, Benedito-Palos L, Estensoro I, Ballester-Lozano G, et al. (2013) Deep sequencing for de novo construction of a marine fish (Sparus aurata) transcriptome database with a large coverage of protein-coding transcripts. BMC Genomics 14: 178. doi: 10.1186/1471-2164-14-178
[46]
Attwood TK, Blythe MJ, Flower DR, Gaulton A, Mabey J, et al. (2002) PRINTS and PRINTS-S shed light on protein ancestry. Nucleic acids research 30: 239–241. doi: 10.1093/nar/30.1.239
[47]
Sigrist CJ, Cerutti L, de Castro E, Langendijk-Genevaux PS, Bulliard V, et al. (2010) PROSITE, a protein domain database for functional characterization and annotation. Nucleic acids research 38: D161–D166. doi: 10.1093/nar/gkp885
[48]
Punta M, Coggill PC, Eberhardt RY, Mistry J, Tate J, et al. (2012) The Pfam protein families database. Nucleic acids research 40: D290–D301. doi: 10.1093/nar/gkr1065
[49]
Salem M, Vallejo RL, Leeds TD, Palti Y, Liu S, et al. (2012) RNA-Seq identifies SNP markers for growth traits in rainbow trout. PLoS One 7: e36264. doi: 10.1371/journal.pone.0036264