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Development of 101 novel EST-derived single nucleotide polymorphism markers for Zhikong scallop (Chlamys farreri)

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Abstract

Zhikong scallop (Chlamys farreri) is an important maricultured species in China. Many researches on this species, such as population genetics and QTL fine-mapping, need a large number of molecular markers. In this study, based on the expressed sequence tags (EST), a total of 300 putative single nucleotide polymorphisms (SNPs) were selected and validated using high resolution melting (HRM) technology with unlabeled probe. Of them, 101 (33.7%) were found to be polymorphic in 48 individuals from 4 populations. Further evaluation with 48 individuals from Qingdao population showed that all the polymorphic loci had two alleles with the minor allele frequency ranged from 0.046 to 0.500. The observed and expected heterozygosities ranged from 0.000 to 0.925 and from 0.089 to 0.505, respectively. Fifteen loci deviated significantly from Hardy-Weinberg equilibrium and significant linkage disequilibrate was detected in one pair of markers. BLASTx gave significant hits for 72 of the 101 polymorphic SNP-containing ESTs. Thirty four polymorphic SNP loci were predicted to be non-synonymous substitutions as they caused either the change of codons (33 SNPs) or pretermination of translation (1 SNP). The markers developed can be used for the population studies and genetic improvement on Zhikong scallop.

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References

  • Choi, Y. S., Lee, K. S., and Park, D. H., 2005. Single nucleotide polymorphism (SNP) detection using microelectrode biochip array. Journal of Micromechanics and Microengineering, 15(10): 1938.

    Article  Google Scholar 

  • Dames, S., Margraf, R. L., Pattison, D. C., Wittwer, C. T., and Voelkerding, K. V., 2007. Characterization of aberrant melting peaks in unlabeled probe assays. Journal of Molecular Diagnostics, 9(3): 290–296.

    Article  Google Scholar 

  • Garvin, M. R., Saitoh, K., and Gharrett, A. J., 2010. Application of single nucleotide polymorphisms to non-model species: a technical review. Molecular Ecology Resources, 10: 915–934.

    Article  Google Scholar 

  • Guo, X., Ford, S. E., and Zhang, F., 1999. Molluscan auqaculture in China. Journal of Shellfish Research, 18(1): 19–31.

    Google Scholar 

  • Jiang, G. D., Li, J. Q., Li, L., Zhang, L. L., and Bao, Z. M., 2011. Development of 44 gene-based SNP markers in Zhikong scallop, Chlamys farreri. Conservation Genetic Resources, 3: 659–663.

    Article  Google Scholar 

  • Li, H., Zhu, D., Gao, X., Li, Y., Wang, J., and He, C., 2010. Mining single nucleotide polymorphisms from EST data of hard clam Meretrix meretrix. Conservation Genetic Resources, 2: 69–72.

    Article  Google Scholar 

  • Liew, M., Seipp, M., Durtschi, J., Margraf, R. L., Dames, S., Erali, M., Voelkerding, K., and Wittwer, C., 2007. Closedtube SNP genotyping without labeled probes/ a comparison between unlabeled probe and amplicon melting. American Jounal of Clinical Pathology, 127: 341–348.

    Article  Google Scholar 

  • Liu, W. D., Li, H. J., Bao, X. B., He, C. B., Li, W. J., and Shan, Z. G., 2011. The first set of EST-derived single nucleotide polymorphism markers for Japanese scallop, Patinopecten yessonsis. Journal of the World Aquaculture Society, 42(3): 456–461.

    Article  Google Scholar 

  • Liu, Z., Karsi, A., and Dunham, R. A., 1999. Development of polymorphic EST markers suitable for genetic linkage mapping of Catfish. Marine Biotechnology, 1(5): 437–447.

    Article  Google Scholar 

  • Moen, T., Hayes, B., Nilsen, F., Delghandi, M., Fjalestad, K. T., Fevolden, S. E., Berg, P. R., and Lien, S., 2008. Identification and characterization of novel SNP markers in Atlantic cod: Evidence for directional selection. BMC Genetics, 9: 18.

    Article  Google Scholar 

  • Muchero, W., Ehlers, J. D., Close, T. J., and Roberts, P. A., 2011. Genic SNP markers and legume synteny reveal candidate genes underlying QTL for Macrophomina phaseolina resistance and maturity in cowpea (Vigna unguiculata (L) Walp). BMC Genomics, 12: 8.

    Article  Google Scholar 

  • Pérez, F., Oritiz, J., Zhinaula, M., Gonzabay, C., Calderón, J., and Volckaert, F. A., 2005. Development of EST-SSR Markers by Data Mining in Three Species of Shrimp: Litopenaeus vannamei, Litopenaeus stylirostris, and Trachypenaeus birdy. Marine Biotechnology, 7(5): 554–569.

    Article  Google Scholar 

  • Qi, H. G., Liu, X., and Zhang, G. F., 2008. Characterization of 12 single nucleotide polymorphisms (SNPs) in Pacific abalone Haliotis discus hannai. Molecular Ecology Resources, 8: 974–976.

    Article  Google Scholar 

  • Qi, H. G., Liu, X., Zhang, G. F., and Wu, F. C., 2009. Mining expressed sequences for single nucleotide polymorphisms in Pacific abalone Haliotis discus hannai. Aquaculture Research, 40: 1661–1667.

    Article  Google Scholar 

  • Sambrook, J., Fritsch, E. F., and Maniatis, T., 1989. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, New York, 467–468.

    Google Scholar 

  • Sauvage, C., Bierne, N., Lapègue, S., and Boudry, P., 2007. Single nucleotide polymorphisms and their relationship to codon usage bias in the Pacific oyster Crassostrea gigas. Gene, 406: 13–22.

    Article  Google Scholar 

  • Syvanen, A. C., 2001. Accessing genetic variation: genotyping single nucleotide polymorphisms. Nature Review Genetics, 2(12): 930–942.

    Article  Google Scholar 

  • Vera, M., Alvarez-Dios, J. A., Millan, A., Pardo, B. G., Bouza, C., Hermida, M., Fernandez, C., Herran, R. D. L., Molina-Luzon, M. J., and Martinez, P., 2011. Validation of single nucleotide polymorphism (SNP) markers from an immune expressed sequence tag (EST) turbot, Scophthalmus maximus, database. Aquaculture, 313: 31–41.

    Article  Google Scholar 

  • Vera, M., Pardo, B. G., Pino-Querido, A., Alvarez-Dios, J. A., Fuentes, J., and Martinez, P., 2010. Characterization of singlenucleotide polymorphism markers in the Mediterranean mussel, Mytilus galloprovincialis. Aquaculture Research, 41: 568–575.

    Article  Google Scholar 

  • Wang, S., Sha, Z., Sonstegard, T., Liu, H., Xu, P., Somridhivei, B., Peratman, E., Kucuktas, H., and Liu, Z., 2008. Quality assessment parameters for EST-derived SNPs from catfish. BMC Genomics, 9: 450.

    Article  Google Scholar 

  • Wang, S., Zhang, L. L., Meyer, E., and Matz, M. V., 2009. Construction of a high-resolution genetic linkage map and comparative genome analysis for a reef-building coral Acropora millepora. Genome Biology, 10: R126.

    Article  Google Scholar 

  • Wang, Y., and Guo, X., 2007. Development and characterization of EST-SNP markers in the Eastern oyster Crassostrea virginica. Marine Biotechnology, 9(4): 500–511.

    Article  Google Scholar 

  • Williams, L. M., Ma, X., Boyko, A. R., Butamante, C. D., and Oleksiak, M. F., 2010. SNP identification, verification, and utility for population genetics in a non-model genus. BMC Genetics, 11: 32.

    Article  Google Scholar 

  • Zhou, L., Myers, A. N., Vandersteen, J. G., Wang, L., and Wittwer, C. T., 2004. Closed-tube genotyping with unlabeled oligonucleotide probes and a saturating DNA dye. Clinical Chemistry, 50(8): 1328–1335.

    Article  Google Scholar 

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Correspondence to Xiaoli Hu.

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Li, J., Bao, Z., Li, L. et al. Development of 101 novel EST-derived single nucleotide polymorphism markers for Zhikong scallop (Chlamys farreri). J. Ocean Univ. China 12, 403–412 (2013). https://doi.org/10.1007/s11802-013-2007-1

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  • DOI: https://doi.org/10.1007/s11802-013-2007-1

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