Genetic Diversity of <i>Dacrycarpus imbricatus</i> At Bukit Tapak, Tabanan, Bali Based on RAPD Marker




Bukit Tapak, Dacrycarpus imbricatus, polimorfisme, RAPD


Dacrycarpus imbricatus Blume is a member of the Podocarpaceae family. In Bali, D. imbricatus was found in Bukit Tapak, Tabanan Regency. This species is one of the dominant species in Bukit Tapak. This study aimed to determine the genetic variation of D. imbricatus in Bukit Tapak using molecular markers RAPD (Random Amplified Polymorphic DNA). The genetic diversity of D. imbricatus needs to be studied to obtain the information used for the conservation of this species. Leaf samples were taken from Bukit Tapak, Candikuning, Baturiti, Tabanan Regency, Bali. DNA was extracted using the CTAB method, followed by extraction using chloroform: isoamyl alcohol. DNA precipitation was carried out using ethanol. RAPD analysis was performed using polymerase chain reaction (PCR) using four primers. PCR products were visualized using agarose gel electrophoresis and ethidium bromide staining. The results showed that the amplified DNA bands ranged from 1 to 5 bands with DNA band sizes ranging from 230 bp - 422 bp. Only OPA4 and UBC106 primers can be used to detect D. imbricatus diversity based on the H, D, and R values. The detected genetic variation is low, as indicated by an average polymorphism of 32.5% and similarities between samples ranging from 0.51 to 1

Author Biographies

Made Pharmawati, Universitas Udayana

Biology Department, Faculty of Mathematics and Natural Sciences, Universitas Udayana, Kampus Bukit Jimbaran, Bali

I Made Anom Sutrisna Wijaya, Universitas Udayana

Department of Biosystems and Agricultural Engineering, Faculty of Agricultural Technology, Universitas Udayana, Kampus Bukit Jimbaran, Bal


Thomas P (2013) Dacrycarpus imbricatus. The IUCN Red List of Threatened Species. e.T42445A2980614.

Fardila D, Sutomo (2011) Efek tepi koridor jalan di Hutan Bukit Pohen, Cagar Alam Batukahu, Bali. Berkala Penelitian.Hayati 17: 9–13

Sutomo, Erosi NK, Undaharta, Bangun TM, Lugrayana IN (2012) Studi awal komposisi dan dinamika vegetasi pohon,hutan Gunung Pohen Cagar Alam Batukahu Bali. Bumi Lestari 12(2): 366-381

van Steenis CGGJ (1972) The mountain flora of Java. E. J. Brill, Leiden

vanZonneveld M, Scheldeman X, Escribano P, Viruel MA, van-Damme P, Garcia W, Tapia C, Romero J, Siguen M, Hormaza IJ (2012) Mapping Genetic Diversity of Cherimoya (Annona cherimola Mill.): Application of spatial analysis for conservation and use of plant genetic resources. PLoS ONE 7(1):e29845. doi:10.1371/journal.pone.0029845

Martin MA, Herrera MA, Martín LM (2012) In situ conservation and landscape genetics in forest species. Journal of Natural Resources and Development 2: 1-5

Nonić M., Å ijaÄić-Nikolić M. (2019) Genetic diversity: Sources, threats, and conservation. In: Leal Filho W., Azul A., Brandli L., Özuyar P., Wall T. (Eds) Life on land. Encyclopedia of the UN Sustainable Development Goals. Springer Cham, Switzerland.

Choudhary N, Ahuja U, Chawla V, Jain RK, Kumari P, Batan KR. (2011) Morphological and molecular variability in weedy rices of Haryana. Asian Journal of Agricultural Research. 5: 250-259

Welsh J, M. McClelland M (1990) Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Research 18(24):7213-7218

Doyle JJ, Doyle JL (1990) A rapid total DNA preparation procedure for fresh plant tissue. Focus 12: 13-15

Pharmawati M, Yan G. McFarlane I. J. (2004) Application of RAPD and ISSR markers to analyse molecular relationships in Grevillea (Proteaceae). Australian Systematic Botany 17: 49-61.

Sambrook J. Fritsch EF, Maniatis T (1989). Molecular Cloning. A Laboratory Manual.: Cold Spring Harbor, Lab.Press,USA

Amiryousefi A, Hyvönen J, Poczai P. (2018) iMEC: Online marker efficiency calculator. Application in Plant Science 6(6):e01159. doi: 10.1002/aps3.1159.

Rohlf F.J (2000). NTSYSpc. Numerical taxonomy and multivariate analysis system. Version 2.1. Exeter Publishing Setauket, New York.

Sahu SK, Thangaraj M, Kathiresan K (2012) DNA extraction protocol for plants with high levels of secondary metabolites and polysaccharides without using liquid nitrogen and phenol. ISRN Molecular Biology 2012: 205049,. doi:10.5402/2012/205049

Aboul-Maaty NAF, Oraby HAS (2019). Extraction of high-quality genomic DNA from different plant orders applying a modified CTAB-based method. Bulletin of the National Research Center 43: 25

Jain SA, de Jesus FT, Marchioro GM, de Araújo, ED (2013). Extraction of DNA from honey and its amplification by PCR for botanical identification. Food Science and Technology 33(4): 753-756.

El-Ashram S, Al Nasr I, Suo, X. (2016). Nucleic acid protocols: Extraction and optimization. Biotechnology reports (Amsterdam, Netherlands) 12: 33–39.

Probojati RT, Wahyudi D, Hapsari L (2019). Clustering analysis and genome inference of pisang raja local cultivars (Musa spp.) from Java Island by Random Amplified Polymorphic DNA (RAPD) Marker. Journal of Tropical Biodiversity and Biotechnology 4(2): 42- 53

Satyawan D, Tasma IM (2011) Genetic diversity analysis of Jatropha curcas provenances using Randomly Amplified Polymorphic DNA Markers Jurnal AgroBiogen 7(1):47-55