Genetic Characteristics of Chloropsis cochinchinensis Gmelin, 1789 Based on The Mitochondrial DNA COI Gene

Jarulis Jarulis, Apin Saputra, R. H. Wibowo
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Abstract

The rate of illegal poaching of blue-winged leafbirds (Chloropsis cochinchinensis) throughout Indonesia, particularly Bengkulu, is quite high. However, only minimal molecular information is available for this species. We performed mtDNA COI gene sequencing to explore genetic characters (conservative site, variable site, parsimony site, and singleton site) of blue-winged leafbirds. Using Qiagen's DNeasy® Blood and Tissue Kit based on the Spin-Column Protocol, total DNA was isolated, and PCR amplification methods were performed. DNA derived from a PCR reaction was forwarded to PT. First Base Malaysia for sequencing. Using MEGA 10.0 and BIOEDIT, the COI gene nucleotide sequence data were assembled, edited, and analyzed to explore of single nucleotide polymorphism, genetic distance, and phylogeny. The 616 bp COI genes contained 566 conservative sites (C), 50 variation sites (V), 24 information parsimony sites (Pi), and 26 singleton sites (S), as indicated by the results. The greatest nucleotide base composition was cytosine (34.1–34.9%), while the lowest was guanine (15.7–16.2%). The proportion of adenine-thymine nucleotide base pairs was greater than that of guanine-cytosine (50.3%). There were 26 barcode-specific mutation sites, 17 transition substitution mutation sites, and 9 transverse substitution mutation sites. The average genetic distance between C. cochinchinensis individuals was 2.2%, but the average genetic difference between species was 9.0%. All C. cochinchinensis individuals in our sample clustered within the same clade and were distinguished from other species within the same genus. The COI gene sequences of C. cochinchinensis that we acquired are novel and can be utilized for molecular identification of the species.
基于线粒体 DNA COI 基因的 Chloropsis cochinchinensis Gmelin, 1789 的遗传特征
在印度尼西亚各地,尤其是明古鲁省,非法偷猎蓝翅大叶鸟(Chloropsis cochinchinensis)的比例相当高。然而,关于该物种的分子信息却少之又少。我们进行了 mtDNA COI 基因测序,以探索蓝翅大叶鸟的遗传特征(保守位点、可变位点、解析位点和单子位点)。使用Qiagen公司的DNeasy®血液和组织试剂盒(基于旋转柱协议)分离总DNA,并进行PCR扩增。从 PCR 反应中获得的 DNA 将转交给 PT.First Base Malaysia 公司进行测序。利用 MEGA 10.0 和 BIOEDIT,对 COI 基因核苷酸序列数据进行了组装、编辑和分析,以探索单核苷酸多态性、遗传距离和系统发育。结果显示,616 bp的COI基因包含566个保守位点(C)、50个变异位点(V)、24个信息解析位点(Pi)和26个单体位点(S)。核苷酸碱基组成中,胞嘧啶最多(34.1%-34.9%),鸟嘌呤最少(15.7%-16.2%)。腺嘌呤-胸腺嘧啶核苷酸碱基对的比例高于鸟嘌呤-胞嘧啶(50.3%)。条形码特异性突变位点有 26 个,过渡取代突变位点有 17 个,横向取代突变位点有 9 个。C. cochinchinensis个体间的平均遗传距离为2.2%,但物种间的平均遗传差异为9.0%。我们样本中的所有 C. cochinchinensis 个体都聚在同一支系中,并与同属的其他物种区分开来。我们获得的 C. cochinchinensis COI 基因序列具有新颖性,可用于该物种的分子鉴定。
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4 weeks
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