Page 31 - 《水产学报》2025年第5期

郑卫卫，等 水产学报, 2025, 49(5): 059102

Identification of a female-specific DNA marker and development of
genetic sex identification method in Verasper variegatus

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ZHENG Weiwei , XU Wenteng , LIU Yang , CHEN Yadong , YANG Tao , XI Xiaoqing ,
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LIU Zhihong , XU Dong , QIN Bo , CHEN Songlin 1,2*
1. State Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Yellow Sea Fisheries Research Institute,
Chinese Academy of Fishery Sciences, Qingdao 266071, China;
2. Laboratory for Marine Fisheries Science and Food Production Processes,
Qingdao Marine Science and Technology Center, Qingdao 266237, China;
3. Rongcheng Marine Economic Development Center, Weihai 264300, China;
4. East China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Shanghai 200090, China
Abstract: Verasper variegatus, a marine flatfish, holds significant nutritional and economic value and exhibits sexual dimorph-
ism in growth, with females growing faster than the males. However, to date, no rapid, effective, and universal sex-specific
DNA marker has been available for genetic sex identification and sex-controlled breeding in V. variegatus. To address this, we
performed whole genome resequencing on 17 male and 17 female individuals to identify female-specific DNA sequences.
Through PCR amplification, we identified a female-specific DNA marker and established a genetic sex identification method.
Our results revealed 359 potential female-specific DNA sequences following reads mapping, sequencing depth, and coverage
analysis. Primer pairs targeting approximately 150 bp DNA sequences flanking these regions were designed. After two rounds
of PCR verification, a 69 bp insertion was identified as a female-specific marker, accurately distinguishing genetic sex via PCR
amplification and 2% agarose gel electrophoresis. Further validated using 117 individuals from another breeding population
confirmed the accuracy of this marker and method, with genetic sex results aligning with phenotypic determined by gonad his-
tology. The development of this female-specific DNA marker and the genetic sex identification method provides crucial genetic
insights into the sex determination mechanism of V. variegatus and accelerates the implementation of sex-controlled breeding
and all-female breeding techniques.
Key words: Verasper variegatus; female-specific DNA marker; genetic sex identification; genome resequencing
Corresponding author: CHEN Songlin. E-mail: chensl@ysfri.ac.cn

Funding projects: STI2030-Major Projects (2023ZD0405505); Shandong Key R & D Program (2022CXPT002); Central Pub-
lic-interest Scientific Institution Basal Research Fund, CAFS (2023TD19); Taishan Scholar Climbing Project of Shandong
Province; Central Public-interest Scientific Institution Basal Research Fund, ECSFRI, CAFS (2016M08)

中国水产学会主办 sponsored by China Society of Fisheries https://www.china-fishery.cn
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