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Int. J. Mol. Sci. 2014, 15(7), 12952-12958; doi:10.3390/ijms150712952
Article

Molecular Method for Sex Identification of Half-Smooth Tongue Sole (Cynoglossus semilaevis) Using a Novel Sex-Linked Microsatellite Marker

1,2
, 1
 and 1,*
Received: 23 January 2014; in revised form: 28 March 2014 / Accepted: 23 April 2014 / Published: 22 July 2014
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
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Abstract: Half-smooth tongue sole (Cynoglossus semilaevis) is one of the most important flatfish species for aquaculture in China. To produce a monosex population, we attempted to develop a marker-assisted sex control technique in this sexually size dimorphic fish. In this study, we identified a co-dominant sex-linked marker (i.e., CyseSLM) by screening genomic microsatellites and further developed a novel molecular method for sex identification in the tongue sole. CyseSLM has a sequence similarity of 73%–75% with stickleback, medaka, Fugu and Tetraodon. At this locus, two alleles (i.e., A244 and A234) were amplified from 119 tongue sole individuals with primer pairs CyseSLM-F1 and CyseSLM-R. Allele A244 was present in all individuals, while allele A234 (female-associated allele, FAA) was mostly present in females with exceptions in four male individuals. Compared with the sequence of A244, A234 has a 10-bp deletion and 28 SNPs. A specific primer (CyseSLM-F2) was then designed based on the A234 sequence, which amplified a 204 bp fragment in all females and four males with primer CyseSLM-R. A time-efficient multiplex PCR program was developed using primers CyseSLM-F2, CyseSLM-R and the newly designed primer CyseSLM-F3. The multiplex PCR products with co-dominant pattern could be detected by agarose gel electrophoresis, which accurately identified the genetic sex of the tongue sole. Therefore, we have developed a rapid and reliable method for sex identification in tongue sole with a newly identified sex-linked microsatellite marker.
Keywords: Cynoglossus semilaevis; sex identification; sex-linked microsatellite marker (SLM); female-associated allele (FAA); multiplex PCR Cynoglossus semilaevis; sex identification; sex-linked microsatellite marker (SLM); female-associated allele (FAA); multiplex PCR
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Liao, X.; Xu, G.; Chen, S.-L. Molecular Method for Sex Identification of Half-Smooth Tongue Sole (Cynoglossus semilaevis) Using a Novel Sex-Linked Microsatellite Marker. Int. J. Mol. Sci. 2014, 15, 12952-12958.

AMA Style

Liao X, Xu G, Chen S-L. Molecular Method for Sex Identification of Half-Smooth Tongue Sole (Cynoglossus semilaevis) Using a Novel Sex-Linked Microsatellite Marker. International Journal of Molecular Sciences. 2014; 15(7):12952-12958.

Chicago/Turabian Style

Liao, Xiaolin; Xu, Genbo; Chen, Song-Lin. 2014. "Molecular Method for Sex Identification of Half-Smooth Tongue Sole (Cynoglossus semilaevis) Using a Novel Sex-Linked Microsatellite Marker." Int. J. Mol. Sci. 15, no. 7: 12952-12958.



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