Ribogospod. nauka Ukr., 2013; 4(26): 66-74
DOI: https://doi.org/10.15407/fsu2013.04.066
УДК [597-146.3:597-146.5]:597.553.2
THE METHOD OF POLYMERASE CHAIN REACTION FOR SEX DETERMINATION IN HUCHEN (HUCHO HUCHO)
Yu. Rud,
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, Institute of Fisheries of NAAS, Kyiv
V. Filipov,
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, Institute of Fisheries of NAAS, Kyiv
L. Dragan,
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, Institute of Fisheries of NAAS, Kyiv
L. Buchatsky,
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, Institute of Fisheries of NAAS, Kyiv
Purpose. To analyse the nucleotide sequences of salmonids Y chromosome and to determine the fragment for specific primers selection and also to develop the PCR based method for sex determination in huchen H. hucho.
Methodology. Using the ClustalW algorithm in MEGA 5.2, the nucleotide sequences of salmonids Y chromosome were analysed. For developing of method for rapid diagnostic of huchen sex the polymerase chain reaction (PCR) assay was used. Tne nucleotide sequences of amplified products were investigated by sequencing.
Findings. Using PCR assay the method of sex determination in huchen H. hucho was developed. It was shown that specific PCR products in size of 450 nucleotides were visible in huchen males only. In addition we showed that selected primers can be used in sex determination of rainbow trout Oncorhynchus mykiss and this fact is proved the high rate of sdY locus similarity and its wide destribution in salmonids.
Originality. The nucleotide sequences of salmonids Y chromosome were analysed and highly conservative region of sdY locus for specific primers selection, which covers sex-linked marker, was identified.
Practical Value. Rapid sex determination in huchen by the developed method will allow to identify reversal males in process of gormonal sex reversion. At the stage of reversal males screening, this method will allow to identify the genotypic males (XY) in experimental group and discard them because only phenotypic males with XX genotype (reversal males) must be used in the crosses with native femelas for getting of 100 % all-females stock.
Key words: huchen (Hucho hucho), sexually dimorphic locus, PCR, sex reversal
REFERENCES
1. Sacobie, C. F. D. & Benfey, T. J. (2005). Sex differentiation and early gonadal development in brook trout. North American Journal of Aquaculture, 67, 181-186. http://dx.doi.org/10.1577/A04-059.1
2. Haffray, P., Lebegue, E. & Jeu, S. (2009). Genetic determination and temperature effects on turbot Scophthalmus maximus sex differentiation: An investigation using steroid sex-inverted males and females. Aquaculture, 294, 30-36. http://dx.doi.org/10.1016/j.aquaculture.2009.05.004
3. Nakamura, M., Kobayashi, Y. & Miura, S. (2005). Sex change in coral reef fish. Fish Physiology and Biochemestry, 31(2-3), 117-122. http://dx.doi.org/10.1007/s10695-006-7595-x
4. Dunham, R. A. (2004). Aquaculture and fisheries biotechnology: genetic approaches. Wallingford: CABI Publishing. http://dx.doi.org/10.1079/9780851995960.0000
5. Sambrook, J. & Russell, D. W. (2001). Molecular cloning: a laboratory manual 3rdedition. New York: Cold Spring Harbour.
6. Phillips, R. B., Konkol, N. R., Reed, K. M. & Stein, J. D. (2001). Chromosome painting supports lack of homology among sex chromosomes in Oncorhynchus, Salmo, and Salvelinus (Salmonidae). Genetica, 111, 119–123. http://dx.doi.org/10.1023/A:1013743431738
7. Rud, Yu. P., Vladimirsky, I. B. & Buchatsky, L. P. (2011). PCR based method for rapid determination of sex in rainbow trout. Aquaculture of Central and Eastern Europe: present and future. 2nd NACEE Conference, Pontos, Chiсinău, 224-226.
8. Yano, A., Nicol, B. & Jouanno, E. (2013). The sexually dimorphic on the Y-chromosome gene (sdY) is a conserved male-specific Y-chromosome sequence in many salmonids. Evolutionary Applications, 6, 486–496. http://dx.doi.org/10.1111/eva.12032
9. Matsuda, M., Nagahama, Y. & Shinomlya, A. (2002). DMY is a Y-specific DM-domain gene required for male development in the medaka fish. Nature, 417, 559–563. http://dx.doi.org/10.1038/nature751
10. Iturra, P., Lam, N.& Fuente, M. (2001). Characterization of sex chromosomes in rainbow trout and coho salmon using fluorescence in situ hybridization (FISH). Genetica, 111, 125–131. http://dx.doi.org/10.1023/A:1013725717142
11. Devlin, R. H. & Nagahama, Y. (2002). Sex determination and sex differentiation in fish: An overview of genetic, physiological, and environmental influences. Aquaculture, 208, 191–364. http://dx.doi.org/10.1016/S0044-8486(02)00057-1
12. Wuertz, S., Gaillard, S. & Barbisan, F. (2006). Extensive screening of sturgeon genomes by random screening techniques revealed no sex-specific marker. Aquaculture, 258, 685–688. http://dx.doi.org/10.1016/j.aquaculture.2006.03.042