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

文献类型： 外文期刊

作者： Liao, Xiaolin 1 ; Xu, Genbo 1 ; Chen, Song-Lin 1 ;

作者机构： 1.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Key Lab Sustainable Dev Marine Fisheries, Minist Agr, Qingdao 266071, Peoples R China

2.Minist Water Resources, Inst Hydroecol, Key Lab Ecol Impacts Hydraul Projects & Restorat, Wuhan 430079, Peoples R China

3.Chinese Acad Sci, Wuhan 430079, Peoples R China

关键词： Cynoglossus semilaevis;sex identification;sex-linked microsatellite marker (SLM);female-associated allele (FAA);multiplex PCR

期刊名称：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES （ 影响因子：5.923； 五年影响因子：6.132 ）

ISSN： 1422-0067

年卷期： 2014 年 15 卷 7 期

页码：

收录情况： SCI

摘要： 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.

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[1]Isolation of Female-Specific AFLP Markers and Molecular Identification of Genetic Sex in Half-Smooth Tongue Sole (Cynoglossus semilaevis). Chen, Song-Lin,Li, Jing,Deng, Si-Ping,Tian, Yong-Sheng,Wang, Qing-Yin,Zhuang, Zhi-Meng,Sha, Zhen-Xia,Xu, Jian-Yong.

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[7]Molecular characterization and expression profiles of LPXRFa at the brain pituitary-gonad axis of half-smooth tongue sole (Cynoglossus semilaevis) during ovarian maturation. Wang, Bin,Liu, Quan,Liu, Xuezhou,Xu, Yongjiang,Shi, Bao,Wang, Bin,Liu, Xuezhou,Xu, Yongjiang,Shi, Bao,Liu, Quan. 2018

[8]Effects of different dietary DHA: EPA ratios on gonadal steroidogenesis in the marine teleost, tongue sole (Cynoglossus semilaevis). Xu, Houguo,Cao, Lin,Wei, Yuliang,Zhang, Yuanqin,Liang, Mengqing,Liang, Mengqing. 2017

[9]Evaluation of reference genes for quantitative real-time PCR analysis of gene expression during early development processes of the tongue sole (Cynoglossus semilaevis). Ma Qian,Zhuang Zhimeng,Feng Wenrong,Liu Shufang,Tang Qisheng. 2015

[10]Growth differences and differential expression analysis of pituitary adenylate cyclase activating polypeptide (PACAP) and growth hormone-releasing hormone (GHRH) between the sexes in half-smooth tongue sole Cynoglossus semilaevis. Ji, Xiang-Shan,Chen, Song-Lin,Xu, Tian-Jun,Yang, Jing-Feng,Tian, Yong-Sheng,Ji, Xiang-Shan,Jiang, Yun-Liang. 2011

[11]Development and characterization of 60 novel EST-SSR markers in half-smooth tongue sole Cynoglossus semilaevis. Sha, Z-X.,Luo, X-H.,Liao, X-L.,Wang, Q-L.,Chen, S-L.,Wang, S-L.,Wang, S-L.. 2011

[12]Molecular cloning, characterization and expression analysis of Sox9a and Foxl2 genes in half-smooth tongue sole (Cynoglossussemilaevis). Dong Xiaoli,Chen Songlin,Shao Changwei,Dong Xiaoli,Ji Xiangshan. 2011

[13]Molecular characterization of Pod1 during sex development in Chinese tongue sole (Cynoglossus semilaevis). Wang, Linna,Zhu, Ying,Xu, Wenteng,Shao, Changwei,Dong, Zhongdian,Li, Hailong,Cui, Zhongkai,Meng, Liang,Guo, Hua,Tian, Yongsheng,Chen, Songlin,Wang, Linna,Xu, Wenteng,Shao, Changwei,Tian, Yongsheng,Chen, Songlin. 2017

[14]Sensory bases and the mode of feeding behavior of tongue sole Cynoglossus semilaevis. Wang Xinan,Ma Aijun. 2009

[15]Effects of BPA and E2 on expression profiles of genes related to hypothalamic-pituitary-gonadal axis of half-smooth tongue sole Cynoglossus semilaevis. Li Fengling,Li Zhaoxin,Wang Qingyin,Zhai Yuxiu. 2013

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[17]Polymorphic dinucleotide microsatellites in tongue sole (Cynoglossus semilaevis). Liao, X.,Shao, C. W.,Tian, Y. S.,Chen, S. L.. 2007

[18]Genetic variability of half-smooth tongue sole Cynoglossus semilaevis populations using microsatellite markers. Liu Shuaishuai,Ye Naihao,Liu Yunguo,Liu Yunguo,Liu Shuaishuai,Guo Yinghui,Li Junfeng. 2011

[19]Two novel calreticulin-related molecules with microbial binding and phagocytosis enhancing capacity in the half-smooth tongue sole, Cynoglossus semilaevis. Wang, Guanghua,He, Shuwen,Zhang, Min,Jiang, Zengjie,Jiang, Zengjie. 2018

[20]Histopathological Liver and Testis Alterations in Male Half Smooth Tongue Sole (Cynoglossus semilaevis) Exposed to Endocrine Disruptors. Li, Fengling,Yao, Lin,Sun, Weihong,Jiang, Yanhua,Li, Zhaoxin,Zhai, Yuxiu. 2017

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