scholarly journals Karakteristik Molekuler Kelelawar (Microchiroptera), berdasarkan DNA Mitokondria (Gen COI) di Gua Sukabumi dan Sentul Jawa Barat

2021 ◽  
Vol 14 (1) ◽  
pp. 20-28
Author(s):  
Ismayati Afifah ◽  
Dedy Duryadi Solihin ◽  
Arzyana Sunkar

AbstrakCytochrome Oxidase I (COI) merupakan salah satu gen mitokondria untuk membantu konstruksi dari pohon filogeni yang dapat bertindak sebagai gen marker. Gen COI memiliki keakuratan dalam mengidentifikasi spesies dan umumnya digunakan sebagai “DNA Barcoding”. Informasi mengenai karakteristik genetik berdasarkan DNA mitokondria pada kelelawar di Sukabumi dan Sentul belum banyak dilaporkan. Tujuan dari penelitian ini untuk mengetahui keragaman genetik kelelawar berdasarkan DNA mitokondria dengan penanda Cytochrome Oxidase I (COI) sebagai DNA barcoding. Isolasi DNA total dilakukan menggunakan Kit Dneasy® Blood and Tissue Kit cat no 69504 (50) berdasarkan prosedur Spin-Column Protocol dengan modifikasi. Hasil penelitian ini menunjukkan bahwa gen COI telah berhasil mengidentifikasi karakteristik spesies. Dua haplotipe didapatkan dari masing-masing populasi. Berdasarkan barcode DNA menunjukkan populasi Sukabumi merupakan spesies Chaerephon plicatus dengan nilai identitas genetik sebesar 97,08%, sedangkan populasi Sentul menunjukkan perbedaan secara genetik dengan spesies Hipposideros larvatus dengan nilai identitas genetik sebesar 94,85%. Identifikasi secara genetik dengan menggunakan gen COI menunjukkan bahwa kelelawar yang berasal Sukabumi adalah spesies Chaerephon plicatus dengan jarak genetik sebesar 3,1%. Kelelawar yang berasal dari Sentul memiliki kedekatan dengan spesies Hipposideros larvatus namun memiliki jarak genetik sebesar 5,2%. AbstractCytochrome Oxidase I (COI) is one of the mitochondrial genes to help the construction of phylogeny trees that can act as marker genes. The COI gene has accuracy in identifying species and is commonly used as "DNA Barcoding". Information about genetic characteristics based on mitochondrial DNA in bats in Sukabumi and Sentul has not been widely reported. The purpose of this study was to determine the genetic diversity of bats based on Mitochondrial DNA with Cytochrome Oxidase I (COI) markers as DNA barcoding. Total DNA isolation was carried out using the Dneasy® Blood and Tissue Kit paint no. 69504 (50) based on the Spin-Column Protocol procedure with modifications. The results of this study indicate that the COI gene has successfully identified species characteristics. Two haplotypes were obtained from each population. Based on DNA barcodes, the population of Sukabumi is a species of Chaerephon plicatus with a genetic identity value of 97.08%, while the Sentul population shows genetic differences with the Hipposideros larvatus species with a genetic identity value of 94.85%. Genetic identification using the COI gene shows that the bats originating from Sukabumi is a spesies Chaerephon plicatus with a genetic distance of 3.1%. The bats originating from Sentul are closely related to the species Hipposideros larvatus but have a genetic distance of 5.2%.

Jurnal MIPA ◽  
2015 ◽  
Vol 4 (1) ◽  
pp. 93
Author(s):  
Thalita C. P. Sumampow

Copepoda merupakan zooplankton kaya manfaat dengan diversitas yang sangat tinggi dan terdiri dari banyak spesies kriptik. Identifikasi cepat, akurat, dan hemat dapat dilakukan dengan menggunakan teknik DNA Barcoding. Kesuksesan teknik tersebut sangat dipengaruhi oleh penggunaan primer yang tepat. Tujuan penelitian ini adalah untuk menguji kemampuan dua pasang primer universal, yakni LCO1490-HCO2198 dan FF2d-FR1d, mengamplifikasi gen COI Copepoda. Dalam penelitian ini, pasangan primer LCO1490-HCO2198 tidak berhasil mengamplifikasi gen target. Sekuens-sekuens hasil amplifikasi menggunakan pasangan primer FF2d-FR1d diidentifikasi melalui BLAST. Hasil yang diperoleh menunjukan bahwa sekuens-sekuens tersebut memiliki persentase kemiripan sebesar 92% dengan bakteri Pandoraea pnomenusa. Melalui hasil yang didapatkan disimpulkan bahwa kedua pasangan primer universal LCO1490-HCO2198 dan FF2d-FR1d tidak cukup spesifik untuk amplifikasi gen cytochrome oxidase I Copepoda.Copepoda is a very beneficial and highly diverse zooplankton with many cryptic species. A fast, reliable, and affordable identification can be done through DNA Barcoding. The success of this technique is affected by the usage of correct primers. The aim of this research was to test the ability of two universal primer pairs, which were LCO1490-HCO2198 and FF2d-FR1d, amplifying COI gene of Copepoda. In this research, LCO1490-HCO2198 primer pairs weren’t able to amplify COI gene of Copepoda. Sequences which were successfully amplified using FF2d-FR1d primer pairs were identified through BLAST. The result shows that the sequences are 92% similar to bacteria named Pandoraea pnomenusa. It can be concluded that both primer pairs are not specific enough to amplify cytochrome oxidase I gene of Copepoda.


2017 ◽  
Vol 3 (1) ◽  
pp. 13-19
Author(s):  
Perkasa Arian ◽  
I Made Artika ◽  
Syamsul Falah

DNA barcoding has become a useful tool for identifying and confirming of species within a known taxonomic framework. A large-scale effort is underway to barcode all amphibian species using the universally sequenced DNA region, a partial fragment of mitochondrial cytochrome oxidase subunit I (COI). This study was aimed to use DNA barcoding technique to identify and confirm species of Polypedates leucomystax and to analyze their phylogenetic relationship. Samples of Polypedates leucomystax were collected from Campus Area of Bogor Agricultural University. The cytochrome oxidase I gene of 600-700 nucleotides were amplified and observed in agarose gel electrophoresis. Forward sequence (604 base pairs) of COI gene was used for phylogenetic analyses. BLAST analysis against BOLD System database showed 95.75% identity with sequences of Polypedates leucomystax. The pairwise genetic distances of Polypedates leucomystax with Rhacophorus schlegelii, Limnonectes fujianensis, Fejervarya cancrivora, and Bufo melanostictus were 0.274, 0.352, 0.339, 0.339, 0.393, respectively. These results illustrated that the genetic identification is congruence with the morphological identification. Phylogenetic tree analysis showed that the samples were in one clade with other tree frogs. The DNA barcoding technique based on the sequence of COI gene can therefore be used to identify and confirm species of Polypedates leucomystax.


2018 ◽  
Vol 13 (3) ◽  
pp. 191
Author(s):  
Melta Rini Fahmi ◽  
Erma Primanita Hayuningtyas ◽  
Mochammad Zamroni ◽  
Bastiar Nur ◽  
Shofihar Sinansari

Ikan tiger fish (Datnioides sp.) merupakan ikan hias air tawar yang memiliki nilai ekonomis penting. Distribusi populasi ikan ini meliputi Papua, Kalimantan, dan Sumatera, dengan tingkat eksploitasi yang cukup tinggi di dua lokasi terakhir. Penelitian ini dilakukan untuk mendapatkan informasi keragaman genetik ikan tiger fish yang mendiami perairan Kalimantan dan Sumatera. Sebanyak 24 sampel ikan uji dikoleksi dari Sungai Kapuas, Kalimantan Barat dan Sungai Musi, Sumatera Selatan. Penelitian dilakukan dalam dua tahap, tahap pertama yaitu identifikasi molekuler dengan menggunakan DNA barcoding gen cytochrome oxidase 1 (COI), tahap kedua adalah analisis keragaman genetik dengan menggunakan marka DNA mitokondria gen cytochrome b (Cyt b), dan DNA inti gen recombination activating gene (RAG2). Hasil identifikasi secara molekuler menunjukkan bahwa ikan hasil koleksi memiliki kesamaan genetik sebesar 100% dengan spesies D. undecimradiatus. Keragaman genetik ikan tiger fish antar populasi berkisar pada nilai 0,023 (standar deviasi 0,001) sedangkan keragaman intra populasi adalah sebesar 0,002 dan 0,003 masing-masing untuk populasi Kalimantan dan Sumatera. Jarak genetik sampel baik yang berasal dari Sumatera maupun Kalimantan dengan spesies D. undeciumradiatus masing-masing 0,003 dan 0,006; sedangkan dengan spesies D. microlepis yaitu 0,142. Analisis menggunakan gen RAG2 menunjukkan sampel yang diuji memiliki struktur populasi yang terpisah ditandai dengan terjadinya mutasi pada enam nukleotida dan tiga asam amino.The Tiger fish (Datnioides sp.) is a freshwater ornamental fish that has important economic value. The distribution of this fish included Papua, Kalimantan, and Sumatra, but intensive exploitation occurs in the last two population. This research was conducted to obtain the genetic diversity of tiger fish that inhabited in Kalimantan and Sumatra. A total of 24 fish were collected from Kapuas River, West Kalimantan and Musi River, at Sumatra. The study was conducted in two stages, the first stage is molecular identification of sample by using DNA barcoding cytochrome oxidase 1 (COI) gene, the second stage is analyses of genetic diversity of tiger fish within and between population by using the mitochondrial DNA cytochrome b (Cyt b) gene, and nucleus DNA recombination (RAG2) gene. The molecular identification has shown that the collected fish has a genetic similarity of 100% with D. undecimradiatus. The genetic diversity of tiger fish between populations is 0.023 (standard deviation of 0.001) whereas intra-population is 0.002 and 0.003 for Kalimantan and Sumatra, respectively. The genetic distance of samples with species D. undeciumradiatus were 0.003 and 0.006 for Kalimantan and Sumatera, respectively, whereas the genetic distance with D. microlepis was 0.142. The analysis of mutation on RAG2 gene shows there are six nucleotides and three amino acids have mutation.


2018 ◽  
Vol 19 (3) ◽  
pp. 997-1003
Author(s):  
R. SUSANTI ◽  
RETNO SRI ISWARI ◽  
FIDIA FIBRIANA ◽  
INDRIAWATI INDRIAWATI

Susanti R, Iswari RS, Fibriana F, Indriawati. 2018. The duck cytochrome oxidase I (COI) gene: Sequence and patterns analysis for potential barcoding tool. Biodiversitas 19: 997-1003. The local duck DNA barcoding is still rarely conducted in Indonesia while DNA barcoding is extensively used as a tool of species identification and delineation tool. This study aimed to analyze the sequence and patterns of Central Javanese ducks mitochondrial cytochrome c oxidase subunit 1 (COI) gene. Feather samples of seven breeds of native duck were collected from traditional husbandries in Central Java. The samples were employed for DNA extraction and COI gene amplification. Five haplotypes were obtained from 35 samples, i.e., haplotype A, B, C, D, and E. Also, 9 variable sites with synonym substitution was detected in four nucleotides number 55, 61, 100, and 109; whereas five synonym substitutions were identified in the nucleotides number 36, 48, 51, 66, and 756. In conclusion, this study annotates that COI mtDNA gene is essential for local ducks barcoding system.


2018 ◽  
Author(s):  
Mohamad-Azam Akmal Abu Bakar ◽  
Jeffrine Japning Rovie-Ryan ◽  
Ahmad Ampeng ◽  
Salmah Yaakop ◽  
Shukor Md Nor ◽  
...  

2021 ◽  
Vol 944 (1) ◽  
pp. 012033
Author(s):  
I G W D Dharmawan ◽  
D G Bengen ◽  
I Setyobudiandi ◽  
B Subhan ◽  
I Verawati ◽  
...  

Abstract Nudibranch has high species diversity with complex morphological characters and is challenging to identify at the species level. The lack of knowledge about nudibranchs makes it difficult to identify conventionally using morphological characters. This study aims to identify nudibranchs at the species level using the DNA barcoding method from the mitochondrial cytochrome oxidase 1 (CO1) gen. The results of DNA barcoding using the Cytochrome Oxidase I (COI) gene showed 18 species of 51 samples analyzed. The phylogenetic tree reconstruction revealed 11 main clades belonging to 11 genera. The genetic distance between and within species clearly shows the difference between individuals. Interspecific genetic distance shows the lowest value between species was found between Chromodoris annae and Chromodoris magnifica is 0.075, and the largest genetic distance observed between species Glossodoris rufomarginata and Tritonidae sp is 0.354. This study shows molecular analysis can be used to identify nudibranch up to species level, which will be a source of information in knowing the distribution and the genetic distance.


2011 ◽  
Vol 207 (1-3) ◽  
pp. e64-e65 ◽  
Author(s):  
Qinlai Liu ◽  
Jifeng Cai ◽  
Yadong Guo ◽  
Xinghua Wang ◽  
Yan Gu ◽  
...  

2017 ◽  
Vol 17 (1) ◽  
pp. 59
Author(s):  
Fitri Elisabeth Br. Hasibuan ◽  
Feky R Mantiri ◽  
Rooije R.H Rumende

KAJIAN VARIASI SEKUNES INTRASPESIES DAN FILOGENETIK MONYET HITAM SULAWESI (Macaca nigra) DENGAN MENGGUNAKAN GEN COI ABSTRAKMacaca nigra merupakan salah satu spesies yang endemik dan terancam punah di Sulawesi Utara. Eksploitasi yang berlebihan serta alih fungsi hutan menjadi ancaman bagi spesies ini di alam. Penelitian ini dilakukan untuk mengetahui variasi sekuens intraspesies M. nigra yang berada di Pusat Penyelamatan Satwa Tasikoki Bitung. Analisis sekuens menunjukkan terdapat perbedaan sepuluh pasang basa nukleotida pada urutan sekuens sampel dilokasi yang berbeda. Jarak genetik antara kedua sampel yaitu 0.016. Hasil perhitungan jarak genetik menunjukkan variasi genetik masih berada dalam kisaran variasi intraspesies dengan ambang batas untuk variasi intraspesies yaitu 0.015-0.025. Selain itu, variasi juga ditunjukkan pada sampel dengan kerabat dekatnya yang terdata di GenBank disebabkan karena adanya mutasi sinonim dan mutasi nonsinonim. Analisis filogenetik berdasarkan gen COI (Cytochrome Oxidase-I) menunjukkan bahwa sampel M. nigra yang digunakan dalam penelitian ini berada satu klaster dengan M. nigra yang ada di database dan termasuk ke dalam kelompok Silenus.Kata kunci: Variasi sekuens intraspesies, Gen COI, Macaca nigra, analisis filogenetik. THE STUDY INTRASPECIFIC VARIATION SEQUENCES AND PHYLOGENETIC CELEBES BLACK MACAQUE (Macaca nigra) USING COI GENE ABSTRACTMacaca nigra is listed as one of the endemic species and endangered in North Sulawesi. Exploitation and forest conversion have become threats to this species in the wild. This study was conducted to determine the intraspecific sequence variation of M. nigra located in Tasikoki Wildlife Rescue Center, Bitung. Sequence analysis revealed ten nucleotides differences between these two specimens. Genetic distance for both of specimens is 0.016. The result of genetic distance, the genetic variation between the specimens of M. nigra was still within the range of intraspecific variation. Distance analysis was also conducted by comparing with the close relatives of M. nigra based on BLAST search, which showed range from 0.015-0.025. These differences resulted in both synonymous and nonsynonymous mutation. Phylogenetic analysis using DNA sequences of COI (Cytochrome Oxidase-I) gene revealed that the two specimens of M. nigra claster together with M. nigra sequences that have been deposited in GeneBank. Moreover M. nigra is claster in the silenus group which is in accordance with previous reports.Keywords: Intraspecific Sequence Variation, COI Gene, Macaca nigra, Phylogenetic analysis.


TREUBIA ◽  
2020 ◽  
Vol 47 (2) ◽  
pp. 99-110
Author(s):  
Jarulis Jarulis ◽  
Dedy Duryadi Solihin ◽  
Ani Mardiastuti ◽  
Lilik Budi Prasetyo

The rhinoceros hornbill (Buceros rhinoceros) genetic characteristics consist of nucleotide polymorphisms, haplotypes, genetic distances, and relationships which are important for their conservation effort in Indonesia. We sequenced mitochondrial DNA D-loop hypervariable III fragments from five rhinoceros hornbill individuals at Safari Park Indonesia I and Ragunan Zoo, which were isolated using Dneasy® Blood and Tissue Kit Spin-Column Protocol, Qiagen. D-loop fragment replication was done by PCR technique using DLBuce_F (5'-TGGCCTTTCTCCAAGGTCTA-3') and DLBuce_R (5'-TGAAGG AGT TCATGGGCTTAG-3') primer. Thirty SNP sites were found in 788 bp D-loop sequences of five rhinoceros hornbill individuals and each individual had a different haplotype. The average genetic distance between individuals was 3.09% and all individuals were categorized into two groups (Group I: EC6TS, EC1RG, EC2TS and Group II: EC9TS, EC10TS) with a genetic distance of 3.99%. This result indicated that the two groups were distinct subspecies. The genetic distance between Indonesian and Thai rhinoceros hornbills was 10.76%. Five Indonesian rhinoceros hornbill individuals at Safari Park Indonesia I and Ragunan Zoo probably came from different populations, ancestors, and two different islands. This study can be of use for management consideration in captive breeding effort at both zoos. The D-loop sequence obtained is a useful character to distinguish three rhinoceros hornbill subspecies in Indonesia.  


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