Abstract. Jarulis, Muslim C, Kamilah SN, Utama AF, Permana D, Sari MM, Prayitno AH, Jannah IM. 2021. DNA barcode of Enggano hill myna, Gracula religiosa enganensis (Aves: Sturnidae) based on mitochondrial DNA cytochrome oxidase subunit I. Biodiversitas 22: 1635-1643. The sharp decline of the Enggano hill myna population due to illegal trading and habitat degradation needs to be our concern to prevent this bird from extinction. Taxonomically, Enggano hill myna is referred to as a sub-species, but this has not been confirmed by genetic data. We have sequenced seventeen Enggano hill myna mitochondrial DNA COI genes to describe their genetic identity (barcode), genetic distances, and phylogeny. DNA genome from seventeen blood samples was isolated with DNeasy® Blood and Tissue Kit Qiagen, while PCR amplification was performed using a pair primers, namely COIGRF (5'-TTCTGATTCTTTGGCCATCC-3') and COIGRR (5'-GTTGGAAGGCTTTGCGTTTA-3'). We used Clustal W alignment in MEGA 10.2.2 software to search single nucleotide polymorphisms. Genetic distance was analyzed by using the Kimura 2-parameter, and the phylogenetic tree was reconstructed with Neighbor-Joining models. We found 98.60% conservative sites, 0.69% parsimony sites, and 0.83% singleton sites from the 716 bp sequence. The highest nucleotide composition was cytosine (32.20%), and the lowest was guanine (16.80%), followed by 49% GC content. Seven SNP sites were found in 716 bp COI gene sequences of seventeen individuals. The genetic distance between Enggano hill myna individuals was ranged from 0.0-0.8%, and all Enggano hill myna individuals separated from Chinese and Indian populations in the phylogenetic tree with a genetic distance of 0.9% and 1.1%. Our data suggest that the Enggano hill myna population is still classified as a sub-species. The COI gene sequences that we found can be used to quickly identify this species and are also important to prevent illegal trading in Indonesia.
As one of the most widespread raptors in Asia, Crested Serpent Eagle (Spilornis cheela) are genetically varied. However, due to the limited studies on genetic aspects, the genetic difference based its geographic distribution are difficult to define. A thorough phylogenetic analysis is needed to create a coherent taxonomic definition for the sake of conservation; starts with phylogenetic analysis based on cytochrome-c oxydase subunit I (COI) gene as the standardized genetic marker to define species. This research was aimed to resolve within species phylogenetic diversity and to explain the relationship between species based on COI gene. Total DNA was isolated from blood samples. The BLAST confirmed PCR amplified COI gene fragments were visualized on 0.8 agarose gel. Sequencing was carried out by FirstBase Laboratory Malaysia. Phylogenetic analysis was done using AMOVA and MEGA7 software in Neighbor Joining method with Kimura-2 parameter. Reconstructed phylogenetic tree confirmed the divergence between three samples and another S. cheela from other regions respective to their geographic distributions. There was 0,014 genetic distance between samples and S. cheela and 0,015 between S. cheela and other species among genus Spilornis. This research also revealed that S. cheela and S. holospilus were not significantly varied.
The study aimed to describe the genetic characteristic, single nucleotide polymorphism, genetic distance, and phylogenetic of mountain white-eye Zosterops montanus based on mitochondrial DNA CO1 gene and carried out in March-June 2020. A sample of the bird was taken from Pasar Minggu, Bengkulu city. The blood sample was taken from the pectoral vein, and the DNA genome was isolated using DNeasy® Blood and Tissue Kit based on Spin-Column Protocol procedure, Qiagen. The CO1 gene was amplified using a polymerase chain reaction technique, electrophoresis on 1.2% agarose gel, and visualized with a UV transilluminator. The results showed that the gene size of CO1 mitochondrial DNA from Z. montanus (n=5) is 751 bp, obtained conservative sites (C) 745 sites, variations (V) six sites, the parsimony of information (Pi) four sites, and singleton sites (S) two sites. The highest nucleotide base composition was cytosine (C) and the lowest guanine (G), while the highest nucleotide combined composition of adenine-thymine (AT) and the lowest guanine-cytosine (GC). We found six single nucleotide polymorphisms were evenly distributed between 9-732 sites. Average intraspecific genetic distance was 0.4%, interspecific genetic distance 4%, and between groups 1 and 2 separated from the outgroup (group 3) with a genetic distance of 12.5%.
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