ISSN : 1225-3480
The method for ploidy verifications is necessary for efficient research and development of triploid abalone, Haliotis discus hannai. The various methods such as flow cytometry, karyotyping, and particle size analysis have been used to confirm ploidy. However, this method provides only an approximate estimation of the ploidy level and measuring cellular DNA content is not only time consuming, but also costly and labor intensive. In this study, a quick and accurate method for ploidy verification of triploid and diploid abalone, H. discus hannai, was developed using multiplex polymerase chain reaction (PCR) system for molecular microsatellite markers. The multiplex PCR system including five microsatellites markers from Pacific abalone, H. discus hannai, consisting of di, tri, tetra and pentanucleotide repeat units, is developed. The multiplex PCR was able to verify the ploidy of 14 triploid abalone with 100% accuracy, In this results suggest that multiplex PCR system may be a useful tool in verification of triploid abalone.
Benfey, T.J., and Sutterlin, A.M. (1984) Triploidy induced by heat shock and hydrostatic pressure in Atlantic salmon (Salmo salar L.). Aquaculture, 36: 359-367.
Chang, Y.J. (1993) Identification of blood cells and their physiological functions in the scallop, Patinopecten yessoensis. Journal of Aquaculture, 6: 1-12.
Chevassus, B. (1983) Hybridization in fish. Aquaculture, 33: 245-262.
Hara, M., and Sekino, M. (2005) Genetic difference between Ezo-awabi Haliotis discus hannai and Kuro-awabi Haliotis discus discus population: microsatellite-based population analysis in Japanese abalone. Fisheries Science, 71: 754-766.
Kim, E.J., Kim, S.J., Park C.J., and Nam Y.K. (2019) Characterization of testis-specific serine/threonine kinase 1-like (TSSK1-like) gene and expression patterns in diploid and triploid Pacific abalone (Haliotis discus hannai). PLoS One 14(12): e0226022.
Park, C.J., Kwon, M.S., Kim E.J., and Nam Y.K. (2018) Improvement of cold-shock treatment conditions for triploidy induction in Pacific abalone Haliotis discus hannai. The Korean Journal Malacology, 34(4): 191-200.
Sekino, M., Shigenobu, Y., Nakamura, Y., Sugaya, T., and Hara, M. (2012) Application of massively parallel pyrosequencing to the discovery of nondinucleotide microsatellite: tetra- and pentanucleotide repeat markers for the Pacific abalone (Haliotis discus hannai). Conservation Genet Resour, 4: 939-941.
Slabbert, R., Prins N., and Brink D. (2010) A microsatellite panel for triploid verification in the abalone Haliotis midae. African Journal of Marine Science, 32(2): 259-264.
Stanley, J. G., Allen, S. K. Jr., and Hidu, H. (1981) Polyploidy induced in the American oyster, Crassostrea virginica, with cytochalasin B. Aquaculture, 23: 1-10.
Suenaga, E., and Nakamura, H. (2005) Evolution of tree methods for effective extraction of DNA from human hair. Journal of chromatography B, 820: 137-141.
Tabarini, C. L., (1984) Induced Triploidy in the bay Scallop, Argopecten irradians, and its effect on growth and gametogenesis. Aquaculture, 42: 151-160.
Thorgaard G.H., Rabinovitch, P.S., Shen, M.W., Gall, G.A.E., Propp, J., and Utter, F.M. (1982) Triploid rainbow trout identified by flow cytometry. Aquaculture, 35: 125-135.
Walsh, P.S., Metzger, D.A., Higuchi, R. (1991) Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. Biotechniques, 10: 506-513.
Wolters, W.R., Chrisman C.L., and Libey, G.S. (1982) Erythrocyte nuclear measurements of diploid and triploid channel catfish, Ictalurus punctatus (Rafinesque). Journal of Fish Biology, 20: 253-258.