DNA barcoding on marine invertebrates
Abstract
A large number of species of marine invertebrates that have similar morphology cause misidentification very possible. Molecular species identification is important in taxonomy study accurately. This study aims to determine the identity of Aquatic Papua marine invertebrates with DNA barcoding technique using markers COI gene (Cytochrome c oxidase I). The study identified 29 individuals of marine invertebrates, which came from sea cucumbers (6 samples), lobsters (6 samples), octopus (6 samples), chitons (5 samples), and sea urchins (6 samples). Molecular identification methods consist of DNA isolation, PCR COI gene fragments, sequencing, and DNA sequence analysis using bioinformatics software. The results showed that the sea cucumber came from the Bohadschia marmorata species, the lobster came from the Panulirus versicolor species, the octopus came from the Octopus cyanea species, the chiton came from the Ischnochitonaustralis species, and the sea urchins came from the Tripneustesgratilla species. The similarity of samples and references are in the range of 84.58 to 100.00%. The disparity index, genetic distance, and phylogenetic tree supported this result.
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References
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Aprilia FE, Soewondo A, Widodo, Toha AHA. 2014. Amplifikasi Gen COI dan 16s rRNA dari Invertebrata Laut Plakobranchus ocellatus. Jurnal Biotropika 2(5): 276-278.
Barber PH, Erdmann MV. 2000. Molecular systematic of the Gonodactylidae (Stomatopoda) using mitochondrial cytochrome oxidase C (subunit 1) DNA sequence data. J. Crustacean Biol. 20: 20–36.
Borges LMS, Hollatz C, Lobo J, Cunha AM, Vilela AP, Calado G, Coelho R, Costa AC, Ferreira MSG, Costa MH, Costa FO. 2016. With a little help from DNA barcoding: Investigating the diversity of Gastropoda from the Portuguese coast. Scientific Reports (6): 1–11.
Casiraghi M, Labra M, Ferri E, Galimberti A, De Mattia F. 2010. DNA barcoding: a six-question tour to improve users' awareness about the method. Briefings in bioinformatics 11(4): 440-453.
Chow S, Konishi K, Mekuchi M, Tamaki Y, Nohara K, Takagi M, Niwa K, Teramoto W, Manabe H, Kurogi H, Suzuki S, Ando D, Tadao J, Kiyomoto M, Hirose M, Shimomura M, Kurashima A, Ishikawa T, Kiyomoto S. 2016. DNA barcoding and morphological analyses revealed validity of Diadema clarki Ikeda, 1939 (Echinodermata, Echinoidea, Diadematidae). Zookeys 585: 1-16. doi: 10.3897/zookeys.585.8161. eCollection 2016.
Dailami M, Santi D, Murtihapsari, Abubakar H, Toha AHA. 2018. Genetic analisys of cytochrome oxidase sub unit 1 gene fragment from Cirrhilabrus cf. ryukyuensis (Labridae) from Cenderawasih Bay and Raja Ampat. Jurnal Iktiologi Indonesia 18(3): 209. https://doi.org/10.32491/jii.v18i3.347
Dharmayanti NLPI. 2011. Filogenetika Molekuler: Metode Taksonomi Organisme Berdasarkan Sejarah Evolusi. Wartazoa 21(1).
Dustin P. 2012. Microtubules. Springer Science & Business Media.
Ehrlich H. 2019. Marine biological materials of invertebrate origin. Springer Nature Switzerland AG. https://doi.org/10.1007/978-3-319-92483-0
Ellis RP, Parry H, Spicer JI, Hutchinson TH, Pipe RK, Widdicombe S. 2011. Immunological function in marine invertebrates: Responses to environmental perturbation. Fish & Shellfish Immunology Journal 30. 1209e1222.
Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R. 1994. DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology 3(5): 294-299.
Fuentes-López A, Ruiz C, Galián J, Romera E. 2020. Molecular identification of forensically important fly species in Spain using COI barcodes. Science and Justice 60(3): 293–302.
Galan GL, Mendez NP, Cruz RYD. 2018. DNA bacoding of three selected gastropod species using cytochrome oxidase (COI) gene. Annals of West University of Timisoara, Ser. Biology 21(1): 93–102.
Hall BG. 2013. Building phylogenetic trees from molecular data with MEGA. Molecular Biology and Evolution 30(5): 1229-1235.
Hebert PDN, Cywinska A, Ball SL, Dewaard JR. 2003. Biological identifications through DNA barcodes. Proc. R. Soc. Lond. B. Biol. Sci. 270: 313–321.
Hebert PDN, Penton EH, Burns JM, Janzen DH, Hallwachs W. 2004. Ten Species In One: DNA Barcoding Reveals Cryptic Species In The Neotropical Skipper Butterfly Astraptes fulgerator. PNAS 101(41): 14812-14817.
Hillis DM, Bull JJ. 1993. An empirical test of bootstrapping as a method for assessing confidence in phylogenetic analysis. Systematic Biology 42: 182–192.
Hughes KA, Pescott OL, Peyton J, Adriaens T, Cottier‐Cook EJ, Key G, Rabitsch W, Tricarico E, Barnes DKA, Baxter N, Belchier M, Blake D, Convey P, Dawson W, Frohlich D, Gardiner LM, González‐Moreno P, James R, Malumphy C, Martin S, Martinou AF, Minchin D, Monaco A, Moore N, Morley SA, Ross K, Shanklin J, Turvey K, Vaughan D, Vaux AGC, Werenkraut V, Winfield IJ, Roy HE. 2020. Invasive non‐native species likely to threaten biodiversity and ecosystems in the Antarctic Peninsula region. Global change biology 26(4): 2702–2716. doi: 10.1111/gcb.14938
Karim A, Iqbal A, Akhtar R, Rizwan M, Amar A, Qamar U, Jahan S. 2015. Barcoding of Fresh Water Fishes From Pakistan. Mitochondrial DNA 1- 4.
Kumar S, Gadagkar SR. 2001. Disparity Index: A simple statistic to measure and test the homogeneity of substitution patterns between molecular sequences. Genetics 158: 1321-1327.
Kumar S, Stecher G, Li M, Knyaz C, Tamura K. 2018. MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms. Mol. Biol. Evol. 35(6): 1547–1549.
Kurniasari RD, Soewondo A, Toha AHA. 2014. Identifikasi Synaptula (Echinodermata : Holothuroidea) Raja Ampat Berdasarkan Gen COI. Jurnal Biotropika 2(5): 265-268.
Mangubhai S, Erdmann MV, Wilson JR, Huffard CL, Ballamu F, Hidayat NI, Wen W. 2012. Papuan Bird’s Head Seascape: Emerging threats and challenges in the global center of marine biodiversity. Marine Pollution Bulletin 64(11): 2279–2295.
Metz EC, Robles-Sikisaka R, Vacquier VD. 1998. Nonsynonymous substitution in abalone spermfertilization genes exceeds substitution in intronsand mitochondrial DNA. Proceedings of the National Academy of Sciences of the United States of America 95: 10676 - 10681.
Muchlisin ZA, Thomy Z, Fadli N, Sarong MA, Siti-Azizah MN. 2013. DNA Barcoding of Freswater Fishes from Lake Laut Tawar, Aceh Province, Indonesia. Acta Ichtyiologica Et Piscatoria 43(1): 21-29.
Nikijuluw VPH. 2017. Biodiversity Conservation of Coastal Fish of Indonesia. Paper Presented at “Simposium Nasional Ikan dan Perikanan Pantai”, Organized by: Masyarakat Iktiologi Indonesia (MII). Bogor, 12-13 September 2017.
Nugraha F, Roslim DI, Ardilla YP, Herman. 2014. Analisis Sebagian Sekuen Gen Ferritin2 pada Padi (Oryza sativa L.) Indragiri Hilir, Riau. Biosaintifika Journal of Biology and Biology Education 6(2): 94-103.
Palumbi SR. 1994. Genetic divergence, reproductive isolation, and marine speciation. In Annual Review of Ecology and Systematics (25): 547–572.
Palumbi SR, Benzie J. 1991. Large mitochondrial DNA differences between morphologically similar Penaeid shrimp. Mol Mar Biol Biotechnol. 1991 Sep;1(1):27-34. PMID: 1669002.
Palumbi SR, Martin A, Romano S, McMillan WO, Stice L, Grabowski G. 2002. The simple fool’s guide to PCR version 2. University of Hawaii, 96822, 1–45.
Pranata B, Toha AHA, Kolibongso D. 2020. Genetic Of Panulirus versicolor Lobster In Cendrawasih Bay Papua And Lombok Waters West Nusa Tenggara. Jurnal Enggano 5(2): 2527-5186.
Puillandre N, Strong EE, Bouchet P, Boisselier MC, Couloux A, Samadi S. 2009. Identifying gastropod spawn from DNA barcodes: possible but not yet practicable. Mol Ecol Res. 9: 1311–1321.
Rahayu DA, Nugroho ED, Listyorini D. 2019. DNA Barcoding Ikan Introduksi Khas Telaga Sari, Kabupaten Pasuruan. Biotropika: Journal of Tropical Biology 7(2): 51-62.
Rahayu DA, Jannah M. 2019. Dna Barcode Hewan dan Tumbuhan Indonesia. Jakarta Selatan: Yayasan Inspirasi Ide Berdaya.
Ratnasingham S, Hebert PD. 2007. BOLD: The Barcode of Life Data System. Molecular ecology notes 7(3): 355-364.
Sahriyani N, Widodo, Toha, AHA. 2014. Amplifikasi gen COI (Cytochrome C Oxidase Subunit I) pada holothuroidea yang dikoleksi dari kepulauan Raja Ampat. Biotropika: Journal of Tropical Biology 2 (4): 232-234.
Saitou N, Nei M. 1987. The neighbour-joining method: A new method for reconstructing phylogenetic trees. Molecular Biology and Evolution (4): 406-425.
Saleky D, Setyobudiandi I, Toha AHA, Takdir M, Madduppa HH. 2016. Length-weight relationship and population genetic of two marine gastropods species (Turbinidae: Turbo sparverius and Turbo bruneus) in the Bird Seascape Papua, Indonesia. Biodiversitas, Journal of Biological Diversity 17(1): 208–217.
Schander C, Willasen E. 2005. What can biological barcoding do for marine biology? Mar Biol Res. 1: 79–83.
Taylor HR, Harrist WE. 2012. An emergent science onthe brink of irrelevance: a review of the past 8 years of DNA barcoding. Molecular Ecology Resources 12(3): 377-388.
Thompson JD, Desmond GH, Toby JG. 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research 22(22): 4673-4680.
Tindi M, Mamangkey NGF, Wullur S. 2017. DNA Barcode dan analisis filogenetik molekuler beberapa jenis bivalvia asal perairan Sulawesi Utara berdasarkan gen COI. Jurnal Pesisir dan Laut Tropis 5(2): 32-38
Toha AHA. 2007. Keragaman Genetik Bulu Babi (Echinoidea). Biota. 12(2):131-135.
Toha AHA, Dailami M, Anwar S, Setiawan JB, Jentewo Y, Lapadi I, Madduppa H. 2020. The genetic relationships and indo-pacific connectivity of whale sharks (Rhincodon typus) with particular reference to mitochondrial COI gene sequences from Cendrawasih bay, Papua, Indonesia. Biodiversitas 21(5): 2159–2171. https://doi.org/10.13057/biodiv/d210544
Walsh PS, Metzger DA, Higuchi R. 2013. Chelex 100 as a Medium for Simple Extraction of DNA for PCR-Based Typing from Forensic Material. BioTechniques 54(3): 506–513. https://doi.org/10.2144/000114018
Ward AK, Classen HL, Buchanan FC. 2009. Fishy-egg tainting is recessively inherited when brown-shelled layers are fed canola meal. Poult. Sci. 88(4): 714-721
Ward RD, Holmes BH, O’Hara TD. 2008. DNA barcoding discriminates echinoderm species. Mol Ecol Res. 8: 1202–1211.
Webb KE, Barnes DKA, Clark MS, Bowden DA. 2006. DNA barcoding: a molecular tool to identify Antarctic marine larvae. Deep-Sea Res Part II Trop Stud Ocean 53: 1053–1060.
Wilkinson CR. 2004. Status of coral reefs of the world: 2004. Global Coral Reef Monitoring Network GCRMN, Australian Institute of Marine Science, Townsville. Queensland, Australia.
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. 1990. Basic Local Alignment Search Tool. J. Mol. Biol. (215): 403-410.
Aprilia FE, Soewondo A, Widodo, Toha AHA. 2014. Amplifikasi Gen COI dan 16s rRNA dari Invertebrata Laut Plakobranchus ocellatus. Jurnal Biotropika 2(5): 276-278.
Barber PH, Erdmann MV. 2000. Molecular systematic of the Gonodactylidae (Stomatopoda) using mitochondrial cytochrome oxidase C (subunit 1) DNA sequence data. J. Crustacean Biol. 20: 20–36.
Borges LMS, Hollatz C, Lobo J, Cunha AM, Vilela AP, Calado G, Coelho R, Costa AC, Ferreira MSG, Costa MH, Costa FO. 2016. With a little help from DNA barcoding: Investigating the diversity of Gastropoda from the Portuguese coast. Scientific Reports (6): 1–11.
Casiraghi M, Labra M, Ferri E, Galimberti A, De Mattia F. 2010. DNA barcoding: a six-question tour to improve users' awareness about the method. Briefings in bioinformatics 11(4): 440-453.
Chow S, Konishi K, Mekuchi M, Tamaki Y, Nohara K, Takagi M, Niwa K, Teramoto W, Manabe H, Kurogi H, Suzuki S, Ando D, Tadao J, Kiyomoto M, Hirose M, Shimomura M, Kurashima A, Ishikawa T, Kiyomoto S. 2016. DNA barcoding and morphological analyses revealed validity of Diadema clarki Ikeda, 1939 (Echinodermata, Echinoidea, Diadematidae). Zookeys 585: 1-16. doi: 10.3897/zookeys.585.8161. eCollection 2016.
Dailami M, Santi D, Murtihapsari, Abubakar H, Toha AHA. 2018. Genetic analisys of cytochrome oxidase sub unit 1 gene fragment from Cirrhilabrus cf. ryukyuensis (Labridae) from Cenderawasih Bay and Raja Ampat. Jurnal Iktiologi Indonesia 18(3): 209. https://doi.org/10.32491/jii.v18i3.347
Dharmayanti NLPI. 2011. Filogenetika Molekuler: Metode Taksonomi Organisme Berdasarkan Sejarah Evolusi. Wartazoa 21(1).
Dustin P. 2012. Microtubules. Springer Science & Business Media.
Ehrlich H. 2019. Marine biological materials of invertebrate origin. Springer Nature Switzerland AG. https://doi.org/10.1007/978-3-319-92483-0
Ellis RP, Parry H, Spicer JI, Hutchinson TH, Pipe RK, Widdicombe S. 2011. Immunological function in marine invertebrates: Responses to environmental perturbation. Fish & Shellfish Immunology Journal 30. 1209e1222.
Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R. 1994. DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology 3(5): 294-299.
Fuentes-López A, Ruiz C, Galián J, Romera E. 2020. Molecular identification of forensically important fly species in Spain using COI barcodes. Science and Justice 60(3): 293–302.
Galan GL, Mendez NP, Cruz RYD. 2018. DNA bacoding of three selected gastropod species using cytochrome oxidase (COI) gene. Annals of West University of Timisoara, Ser. Biology 21(1): 93–102.
Hall BG. 2013. Building phylogenetic trees from molecular data with MEGA. Molecular Biology and Evolution 30(5): 1229-1235.
Hebert PDN, Cywinska A, Ball SL, Dewaard JR. 2003. Biological identifications through DNA barcodes. Proc. R. Soc. Lond. B. Biol. Sci. 270: 313–321.
Hebert PDN, Penton EH, Burns JM, Janzen DH, Hallwachs W. 2004. Ten Species In One: DNA Barcoding Reveals Cryptic Species In The Neotropical Skipper Butterfly Astraptes fulgerator. PNAS 101(41): 14812-14817.
Hillis DM, Bull JJ. 1993. An empirical test of bootstrapping as a method for assessing confidence in phylogenetic analysis. Systematic Biology 42: 182–192.
Hughes KA, Pescott OL, Peyton J, Adriaens T, Cottier‐Cook EJ, Key G, Rabitsch W, Tricarico E, Barnes DKA, Baxter N, Belchier M, Blake D, Convey P, Dawson W, Frohlich D, Gardiner LM, González‐Moreno P, James R, Malumphy C, Martin S, Martinou AF, Minchin D, Monaco A, Moore N, Morley SA, Ross K, Shanklin J, Turvey K, Vaughan D, Vaux AGC, Werenkraut V, Winfield IJ, Roy HE. 2020. Invasive non‐native species likely to threaten biodiversity and ecosystems in the Antarctic Peninsula region. Global change biology 26(4): 2702–2716. doi: 10.1111/gcb.14938
Karim A, Iqbal A, Akhtar R, Rizwan M, Amar A, Qamar U, Jahan S. 2015. Barcoding of Fresh Water Fishes From Pakistan. Mitochondrial DNA 1- 4.
Kumar S, Gadagkar SR. 2001. Disparity Index: A simple statistic to measure and test the homogeneity of substitution patterns between molecular sequences. Genetics 158: 1321-1327.
Kumar S, Stecher G, Li M, Knyaz C, Tamura K. 2018. MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms. Mol. Biol. Evol. 35(6): 1547–1549.
Kurniasari RD, Soewondo A, Toha AHA. 2014. Identifikasi Synaptula (Echinodermata : Holothuroidea) Raja Ampat Berdasarkan Gen COI. Jurnal Biotropika 2(5): 265-268.
Mangubhai S, Erdmann MV, Wilson JR, Huffard CL, Ballamu F, Hidayat NI, Wen W. 2012. Papuan Bird’s Head Seascape: Emerging threats and challenges in the global center of marine biodiversity. Marine Pollution Bulletin 64(11): 2279–2295.
Metz EC, Robles-Sikisaka R, Vacquier VD. 1998. Nonsynonymous substitution in abalone spermfertilization genes exceeds substitution in intronsand mitochondrial DNA. Proceedings of the National Academy of Sciences of the United States of America 95: 10676 - 10681.
Muchlisin ZA, Thomy Z, Fadli N, Sarong MA, Siti-Azizah MN. 2013. DNA Barcoding of Freswater Fishes from Lake Laut Tawar, Aceh Province, Indonesia. Acta Ichtyiologica Et Piscatoria 43(1): 21-29.
Nikijuluw VPH. 2017. Biodiversity Conservation of Coastal Fish of Indonesia. Paper Presented at “Simposium Nasional Ikan dan Perikanan Pantai”, Organized by: Masyarakat Iktiologi Indonesia (MII). Bogor, 12-13 September 2017.
Nugraha F, Roslim DI, Ardilla YP, Herman. 2014. Analisis Sebagian Sekuen Gen Ferritin2 pada Padi (Oryza sativa L.) Indragiri Hilir, Riau. Biosaintifika Journal of Biology and Biology Education 6(2): 94-103.
Palumbi SR. 1994. Genetic divergence, reproductive isolation, and marine speciation. In Annual Review of Ecology and Systematics (25): 547–572.
Palumbi SR, Benzie J. 1991. Large mitochondrial DNA differences between morphologically similar Penaeid shrimp. Mol Mar Biol Biotechnol. 1991 Sep;1(1):27-34. PMID: 1669002.
Palumbi SR, Martin A, Romano S, McMillan WO, Stice L, Grabowski G. 2002. The simple fool’s guide to PCR version 2. University of Hawaii, 96822, 1–45.
Pranata B, Toha AHA, Kolibongso D. 2020. Genetic Of Panulirus versicolor Lobster In Cendrawasih Bay Papua And Lombok Waters West Nusa Tenggara. Jurnal Enggano 5(2): 2527-5186.
Puillandre N, Strong EE, Bouchet P, Boisselier MC, Couloux A, Samadi S. 2009. Identifying gastropod spawn from DNA barcodes: possible but not yet practicable. Mol Ecol Res. 9: 1311–1321.
Rahayu DA, Nugroho ED, Listyorini D. 2019. DNA Barcoding Ikan Introduksi Khas Telaga Sari, Kabupaten Pasuruan. Biotropika: Journal of Tropical Biology 7(2): 51-62.
Rahayu DA, Jannah M. 2019. Dna Barcode Hewan dan Tumbuhan Indonesia. Jakarta Selatan: Yayasan Inspirasi Ide Berdaya.
Ratnasingham S, Hebert PD. 2007. BOLD: The Barcode of Life Data System. Molecular ecology notes 7(3): 355-364.
Sahriyani N, Widodo, Toha, AHA. 2014. Amplifikasi gen COI (Cytochrome C Oxidase Subunit I) pada holothuroidea yang dikoleksi dari kepulauan Raja Ampat. Biotropika: Journal of Tropical Biology 2 (4): 232-234.
Saitou N, Nei M. 1987. The neighbour-joining method: A new method for reconstructing phylogenetic trees. Molecular Biology and Evolution (4): 406-425.
Saleky D, Setyobudiandi I, Toha AHA, Takdir M, Madduppa HH. 2016. Length-weight relationship and population genetic of two marine gastropods species (Turbinidae: Turbo sparverius and Turbo bruneus) in the Bird Seascape Papua, Indonesia. Biodiversitas, Journal of Biological Diversity 17(1): 208–217.
Schander C, Willasen E. 2005. What can biological barcoding do for marine biology? Mar Biol Res. 1: 79–83.
Taylor HR, Harrist WE. 2012. An emergent science onthe brink of irrelevance: a review of the past 8 years of DNA barcoding. Molecular Ecology Resources 12(3): 377-388.
Thompson JD, Desmond GH, Toby JG. 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research 22(22): 4673-4680.
Tindi M, Mamangkey NGF, Wullur S. 2017. DNA Barcode dan analisis filogenetik molekuler beberapa jenis bivalvia asal perairan Sulawesi Utara berdasarkan gen COI. Jurnal Pesisir dan Laut Tropis 5(2): 32-38
Toha AHA. 2007. Keragaman Genetik Bulu Babi (Echinoidea). Biota. 12(2):131-135.
Toha AHA, Dailami M, Anwar S, Setiawan JB, Jentewo Y, Lapadi I, Madduppa H. 2020. The genetic relationships and indo-pacific connectivity of whale sharks (Rhincodon typus) with particular reference to mitochondrial COI gene sequences from Cendrawasih bay, Papua, Indonesia. Biodiversitas 21(5): 2159–2171. https://doi.org/10.13057/biodiv/d210544
Walsh PS, Metzger DA, Higuchi R. 2013. Chelex 100 as a Medium for Simple Extraction of DNA for PCR-Based Typing from Forensic Material. BioTechniques 54(3): 506–513. https://doi.org/10.2144/000114018
Ward AK, Classen HL, Buchanan FC. 2009. Fishy-egg tainting is recessively inherited when brown-shelled layers are fed canola meal. Poult. Sci. 88(4): 714-721
Ward RD, Holmes BH, O’Hara TD. 2008. DNA barcoding discriminates echinoderm species. Mol Ecol Res. 8: 1202–1211.
Webb KE, Barnes DKA, Clark MS, Bowden DA. 2006. DNA barcoding: a molecular tool to identify Antarctic marine larvae. Deep-Sea Res Part II Trop Stud Ocean 53: 1053–1060.
Wilkinson CR. 2004. Status of coral reefs of the world: 2004. Global Coral Reef Monitoring Network GCRMN, Australian Institute of Marine Science, Townsville. Queensland, Australia.
Published
2021-06-25
How to Cite
HIKAM, Agus Mohammad et al.
DNA barcoding on marine invertebrates.
Jurnal Biologi Udayana, [S.l.], v. 25, n. 1, p. 46-56, june 2021.
ISSN 2599-2856.
Available at: <https://ojs.unud.ac.id/index.php/bio/article/view/70835>. Date accessed: 26 apr. 2024.
doi: https://doi.org/10.24843/JBIOUNUD.2021.v25.i01.p06.
Section
Articles
