DNA Isolation And Amplification of the rbcL (ribulose-1,5- bisphosphate carboxylase/oxygenase large subunit) gene of Caulerpa sp., Gracilaria sp., And Sargassum sp.

Authors

DOI:

https://doi.org/10.35800/jip.8.2.2020.30003

Keywords:

DNA, gene rbcL, algae Caulerpa sp., Sargassum sp., Gracilaria sp

Abstract

DNA isolation and gene amplification of algae are significantly influenced by various factors such as characteristics and components of the algae cell wall. Therefore techniques and methods of DNA isolation in certain algae, sometimes only succeed in one particular species and can not be applied to another algae species. Based on that issue, this study was conducted with the aims to determine the succeed of DNA isolation and amplify the rbcL gene as a target gene for identification. Algae DNA was isolated by using innuPrep Plant DNA commercial kit, and the second one with a modified conventional Cetyl Trimetyl Ammonium Bromide (CTAB) method,  for the amplification process was using rbcL gene (ribulose-1,5-bisphosphate carboxylase / oxygenase large subunit) with two pairs of primers : rbcL 7F-753R and rbcL 577F-rbcSR. The results showed that the DNA of Gracilaria sp was succeed isolated by using CTAB method and it was denoted by the presence of DNA bands in agarose gel. Meanwhile DNA amplification for Gracilaria sp., and Sargassum sp., were succeed amplified with the appearance of DNA bands. But in algae Caulerpa sp., was only succeed on 1 pair of primary rbcL 7F and 7.

Keywords : DNA, gene rbcL, algae Caulerpa sp., Sargassum sp., Gracilaria sp;

Abstrak

Isolasi DNA dan amplifikasi gen pada alga sangat dipengaruhi oleh beberapa faktor seperti karakter dan komponen pada dinding sel alga. Oleh karena itu proses isolasi DNA terkadang bisa berhasil pada satu jenis alga, namun tidak berhasil pada jenis alga lainnya. Oleh karena alasan tersebut, maka penelitian ini dilakukan untuk menentukan keberhasilan Isolasi DNA dan mengamplifikasi gen rbcL sebagai gen target identifikasi. Penelitian ini dilakukan dengan tahapan awal Isolasi DNA yang menggunakan kit komersil innuPrep Plant DNA Kit, dan metode konvensional Cetyl Trimetyl Ammonium Bromide (CTAB) yang telah dimodifikasi. Sedangkan untuk proses amplifikasi, menggunakan gen rbcL (ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit) digunakan dua pasang primer yaitu rbcL 7F-753R dan rbcL 577F-rbcSR. Hasil isolasi DNA dari alga Gracilaria sp berhasil diisolasi menggunakan metode CTAB yang ditandai dengan adanya pita DNA pada gel agarose. Amplifikasi DNA pada alga Gracilaria sp., dan Sargassum sp., berhasil diamplifikasi dengan munculnya pita DNA. Namun pada alga Caulerpa sp. hanya berhasil pada 1 pasang primer rbcL 7F dan753R.

Kata kunci : DNA, gen rbcL, alga Caulerpa sp., Sargassum sp., Gracilaria sp.

References

Alberto, F., R. Santos, & J.M. Leitao. 1997. DNA extraction and RAPD markers to assess the genetic similarity among Gelidium sesquipedale (Rhodophyta) populations. J. Phycology, 33:706-710.

Allen, G. C., M. A. Flores-Vergara, S. Krasynanski, S. Kumar, & W. F. Thompson. 2006. A modified protocol for rapid DNA isolation from plant tissues using cetyltrimethylammonium bromide. Nature Protocols. 1 (5): 2320-2325

Anggraeni, S. R., Sudarsono., & D. Soedharma. 2008. Karakterisasi Genetika Rumput Laut Eucheuma spp. Dari Tiga Daerah di Indonesia (Kepulauan Seribu, Keruak, dan Sumenep). Jurnal Bionatura. Vol 10(3): 196-208

Annisaqois, M., G. S. Gerung., S. Wullur., D. A. Sumilat., B. T. Wagey & S. V. Mandagi. 2018. Analisis Molekuler DNA Alga Merah (RHODOPHYTA) Kappaphycus sp. Jurnal Pesisir dan Laut Tropis. Vol 1(1): Hal 107-112.

Doyle, J.J. & Doyle, J. L. 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem. Bull. Vol 19: Pp 11-15

Doyle, J.J. & Doyle, J. L. 1990. Isolation of plant DNA from fresh tissue. Focus, 12:13.

Geraldino, J.P., Chang, Y.E. and Kim, M.S. 2009. Systematics of Hypnea asiatica sp. nov. (Hypneaceae, Rhodophyta) Based on Morphology and nrDNA SSU, Plastid rbcL, and Mitochondrial Cox1. Taxon, 58(2), 606-616.

Gerung, S.G. 2004. Biodiversity of Indonesian seaweeds. In: Marine Science into the New Millennium: New Perspective & Challenges. Phang, S.M., Chong, V.C., Ho, S.C., Mokhtar, N. and Sim, J.O.L. pp. 41-54.

Gerung,G.S. 2006 Seaweeds from Manado Bay, Indonesia. In Advance in Seaweeds cultivation and untilization in Asia (eds. Phang Siew Mo. Alan T. Critchley & Put O. Ang jr.). University of Malaya Maritime Research Centre, Malaysia. Pp. 35-40.

Greco, M., C. A. Saez., M. T. Brown., & M. B. Bitonti. 2014. A Simple Extraction of Genomic DNA and Total RNA from Low Biomass Ectocarpus siliculosus, the Model Brown Alga. Journal Plos one 9(7): e96470.

Hengkengbala, I. R. G. S. Gerung., S. Wullur. 2018. Ekstraksi DNA dan Amplifikasi gen rbcL (ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit) Alga Merah Gracilaria sp. dari Perairan Desa Bahoi, Kabupaten Minhasa Utara. Journal of Aquatic Science & Management. 6(2): 33-38.

Kadi, A. 2005. Beberapa Catatan Kehadiran Marga Sargassum di Perairan Indonesia. Oseana, 30 (4) : 19-29.

Kumari, B. 2013. DNA Isolation Protokol for Different Types of Seaweeds From Seashore Areas of Rameswaram. Applied Biology and Biotechnology. Vol 1(2): Hal 5-10

Lyra, G. M., Costa, E.S., De Jesus P. B., De Matos, J. C. G. and Caires, T. A. 2015. Phylogeny Of Gracilariaceae (Rhodophyta): Evidence from Plastid and Mitochondrial Nucleotide Sequences. Journal Of Phycology, 51, 356-366

Nontji, A. 1993. Laut Nusantara. Jakarta: Perebit Djambatan. 367 hal

Phillips, N., Smith, C. M., Morden, C. W. 2001. An effective DNA extraction protocol for Brown Algae. Phycological Research, 49, 97-102.

Ruenes, J. 2010. DNA barcoding of Select Freshwater and Marine Red Algae (Rhodophyta). Cryptogamie, Algologie, 31 (4),377-386

Sahri, A. and Suparmi. 2009. Mengenal Potensi Alga : Kajian Pemanfaatan Sumber Daya Alga Dari Aspek Industri Dan Kesehatan. Sultan Agung, XLIV(118).

Shiviji, M. S. Rogers, S. O Stanhope, M. J. 1992. Rapid Isolation of High Moleculer Weight DNA from Marine Macroalgae. Marine Ecology Progress Series. Vol 84: Hal 197-203

Teddy, M.S., 2009, Pembuatan nori Secara Tradisional Dari Rumput Laut Jenis Gracilaria sp.,. Skripsi Program Studi Teknologi Hasil Perairan. Fak. Perik. Dan Ilmu Kelautan. IPB. Hal 3-4

Terada, R., & Watanebe, Y. 2016. Seaweeds and Seagrasses in the Amami Islands: Biodiversity and Utilization. Chapter 17. Kagoshima University Research Center for the Pacific Island. Pp 107-115

Trono, G. C. & E. P. Ganzon-Fortes. 1988. Philippine Seaweeds. Philippines: National Book Store. Inc Publishers Metro Manila. Pp 330

Tuney, I. and Sukatar, A. 2010. DNA extraction protocol from Brown Algae. Biological Diversity and Conservation. Vol 3(1): Pp 51-55.

Wattier, R.A., Prodohl, P.A. and Maggs, C.A. 2000. DNA Isolation Protocol for Red Seaweed (Rhodophyta). Plant Molecular Biology Reporter, 18, 275-281.

Weeden N.F., Timmerman G.M., Hemmat M., Kneen B.E., Lodhi M.A. 1992. Inheritance & Reability of RAPD markers. In Applications of RAPID technology to plant breeding, Symposium Proceedings. Crop Science Society of America, Madison. pp 12-17

Yang, E.C., Kim, M.S., Geraldino, P.J.L., Sahoo, D., Shin, J. and Boo, S.M. 2013. Mitochondrial cox1 and Plastid rbcL Genes of Gracilaria vermiculophylla (Gracilaria-ceae, Rhodophyta). Journal of Applied Phycology, 20, 161-168

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Published

2020-08-14

How to Cite

Tampanguma, B., Gerung, G. S., Warouw, V., Wagey, B. T., Wulllur, S., Sumilat, D. A., & Onibala, H. (2020). DNA Isolation And Amplification of the rbcL (ribulose-1,5- bisphosphate carboxylase/oxygenase large subunit) gene of Caulerpa sp., Gracilaria sp., And Sargassum sp. Jurnal Ilmiah Platax, 8(2), 214–220. https://doi.org/10.35800/jip.8.2.2020.30003

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