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Identification of the male sex-linked DNA sequence of cultured white trevally Pseudocaranx dentex

机译:养殖白鲹Pseudocaranx dentex雄性性别连锁DNA序列的鉴定

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White trevally Pseudocaranx dentex is a high-value carangid fish species that is only cultured in Japan under full lifecycle aquaculture conditions. To make the reproductive process successful and efficient, sexual phenotyping of broodstock is essential, and this has conventionally been practiced via examination upon abdominal compression. However, in certain cases, sexual phenotyping is not successful due to unsuccessful sperm or semen collection, which leads to misclassification of the phenotype. This study identified the male characteristic DNA sequence of P. dentex. The pectoral fins of eight male and eight female 6-year-old F3 generation broodstock fish were utilized for male sex hormone quantification to determine each sex phenotype. The same fish DNA samples were subjected to bulked-amplified fragment length polymorphism (AFLP) screening, and a male characteristic AFLP DNA fragment of 227 bp in size was identified. A male characteristic fragment was also confirmed by second amplification when it was performed individually. All eight males used for screening carried the AFLP fragment, while none of the eight females did. AFLP was conducted for all 40 broodstock, and it was determined that all 24 putative males were AFLP positive, and 4 of the 16 putative females were AFLP positive. These results indicate that P. dentex may have a male characteristic DNA polymorphism. The nucleotide sequence of the male characteristic fragment showed high homology to Atlantic horse mackerel Trachurus trachurus chromosome 5, and the sequence did not have clear homology to known coding genes. Further studies are required to identify the sex-determining gene of P. dentex in the vicinity of the DNA sequence identified in this study.
机译:白鲹 Pseudocaranx dentex 是一种高价值的鲹科鱼类,仅在日本全生命周期水产养殖条件下养殖。为了使繁殖过程成功和高效,亲鱼的性表型是必不可少的,这通常是通过腹部压迫检查来进行的。然而,在某些情况下,由于精子或精液采集不成功,性表型不成功,这会导致表型的错误分类。本研究鉴定了齿状假单胞菌的雄性特征DNA序列。利用8条雄性和8条雌性6年F3代亲鱼的胸鳍进行雄性激素定量,以确定每种性别表型。对相同的鱼类DNA样本进行扩增片段长度多态性(AFLP)筛选,鉴定出大小为227 bp的雄性特征AFLP DNA片段。当单独进行时,男性特征片段也通过第二次扩增得到确认。用于筛查的所有八名男性都携带AFLP片段,而八名女性都没有携带AFLP片段。对所有 40 只亲鱼进行了 AFLP,确定所有 24 只推定雄性均为 AFLP 阳性,16 只推定雌性中有 4 只为 AFLP 阳性。这些结果表明,齿状假单胞菌可能具有雄性特征DNA多态性。雄性特征片段的核苷酸序列与大西洋竹荚鱼Trachurus trachurus 5号染色体具有高度同源性,且该序列与已知编码基因没有明确的同源性。需要进一步的研究来鉴定本研究中鉴定的 DNA 序列附近的齿状假单胞菌的性别决定基因。

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