Molecular Identification of Silver-Barb (Barbonymus sp.) Based on COI and Cyt-b gene region from East Java

Mutiara Rahmawati, A. Mukti, S. Andriyono
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Abstract

This study aims to determine the genetic character of cyprinid fish (Barbonymus sp.) using the mtDNA gene in the COI and Cyt-b gene regions. Samples were collected from the various areas in East Java Province, including Gresik, Mojokerto, Madiun, Malang, and Banyuwangi. The caudal peduncle of the cyprinid fish was used for molecular identification. Isolation and purification of its genomic DNA using the gSYNCTM DNA Extraction kit. We used a polymerase chain reaction machine for template DNA amplification with COI primers, including FishF1 and FishR1, while Cyt-b primers, including GluDG-L and CB2-H. The results showed that all cyprinid fish sequences were successfully amplified with the COI gene of 648 bp and the Cyt b gene of 408 bp. The results of the BLASTN analysis show that there are two species of cyprinid fish that have been found, namely Barbonymus gonionotus and Barbonymus schwanenfeldii, with percentage identities ranging from 99-100%. The species Barbonymus gonionotus was found in five regions (Gresik, Mojokerto, Madiun, Malang, and Banyuwangi), while Barbonymus schwanenfeldii was only found in Malang. The genetic distance value based on the COI gene ranged from 0.00000-0.10694, while the genetic distance value based on the Cyt-b gene ranged from 0.00000-1.15662. Conservation status based on IUCN Red List data, the species Barbonymus gonionotus and Barbonymus schwanenfeldii are included in the Least Concern (LC) category, and these species, according to CITES data, have not been evaluated or Not Evaluated.
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基于东爪哇COI和Cyt-b基因区的银Barb(Barbonymus sp.)分子鉴定
本研究旨在利用COI和Cyt-b基因区的mtDNA基因来确定塞浦路斯鱼(Barbonymus sp.)的遗传特征。样本采集自东爪哇省的各个地区,包括Gresik、Mojokerto、Madiun、Malang和Banyuwangi。利用塞浦路斯鱼的尾鳍进行分子鉴定。使用gSYNCTM DNA提取试剂盒分离和纯化其基因组DNA。我们使用聚合酶链式反应机用COI引物(包括FishF1和FishR1)和Cyt-b引物(包括GluDG-L和CB2-H)进行模板DNA扩增。结果表明,成功地扩增出了所有塞浦路斯鱼类序列,其中COI基因为648bp,Cyt b基因为408bp。BLASTN分析结果表明,目前已发现的塞浦路斯鱼类有两种,分别为卵圆矛和schwanenfeldii,其百分比同一性在99-100%之间。gonionotus Barbonymus在五个地区(Gresik、Mojokerto、Madiun、Malang和Banyuwangi)发现,而schwanenfeldii Barbonymous仅在Malang发现。基于COI基因的遗传距离值在0.00000-0.10694之间,而基于Cyt-b基因的遗传距值在0.00000-1.15662之间。根据国际自然保护联盟红色名录的数据,该物种被列入最不受关注(LC)类别,而根据濒危野生动植物种国际贸易公约的数据,这些物种尚未被评估或未被评估。
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