SEQUENCE ANALYSIS OF 18s DNA OF <i>Melosira</i> sp., <i>Dunaliella</i> sp., <i>Isochrysis</i> sp. AND <i>Porphyridium</i> sp.
Phytoplankton has high level of biodiversity. In previous years phytoplankton was identified by their morphological characters. However, their morphology might change in different environments. These difficulties can be overcome by comparing their 18S rDNA sequences. This research is aimed to verify the identity of Melosira sp., Dunaliella sp., Isochrysis sp. and Porphyridium sp. Here, PCR method was used to amplify 18s DNA sequences. Three primer pairs were used, i.e. 18S-F and 18S-R; 501F and 1700R; 18S-2F and 18S-2R. PCR products were sequenced. MEGA5 was used to make phylogenetic tree. Genus verification for Isochrysis sp., Dunaliella sp. and Melosira sp. were conducted successfully using Blast and phylogenetic tree. 18s DNA sequence of Porphyridium sp. shows an interesting result and needs further verification.
Keywords: Phytoplankton, Melosira sp., Dunaliella sp., Isochrysis sp., Porphyridium sp.
Adda, M., J.C. Merchuk, and S. Arad. 1986. Effect of nitrate on growth and production of cell wall polysaccharide by the unicellular red alga Porphyridium. Biomass 10, 131 – 140.
Brouard, J-S., C. Otis, C. Lemieux, and M. Turmel. 2008. Chloroplast DNA sequence of the green alga Oedogonium cardiacum (Chlorophyceae): Unique genome architecture, derived characters shared with the Chaetophorales and novel genes acquired through horizontal transfer. BioMed Central Genomics 9, 290.
Cavalier-Smith, T. 1998. A revised six-kingdom system of life. Biological Reviews 73, 203 266.
Chen, Y.C., J.D. Eisner, M.M. Kattar, S.L. Rassoulian-Barret, K. Lafe, A.P. Limaye, and B.T. Cookson. 2001. Polymorphic Internal Transcribed Spacer region 1 DNA sequences identify medically important yeast. Journal Clinical Microbiology 39, 4042 – 4051.
Daniel, L.N., and E.S. Douglas. 1995. A comparison of Angiosperm phylogenies from nuclear 18S rDNA and rbcL sequences. Annals of the Missouri Botanical Garden 82, 208 234.
Krienitz, L., I. Ustinova, T. Friedl, and V.A.R. Huss. 2001. Traditional generic concepts versus 18S rRNA gene phylogeny In the green algal family Selenastraceae (Chlorophyceae, Chlorophyta). Journal of Phycology 37, 852 - 865.
Larkin, M.A., G. Blackshields, N.P. Brown, R. Chenna, P.A. McGettigan, H. McWilliam, F. Valentin, I.M. Wallace, A. Wilm, R. Lopez, J.D. Thompson, T.J. Gibson, and D.G. Higgins. 2007. ClustalW and ClustalX version 2. Bioinformatics 23, 2947 – 2948.
Lattès, S., H. Ferté, P. Delaunay, J. Depaquit, M. Vassallo, M. Vittier, S. Kokcha, E. Coulibaly, and P. Marty. 2011. Trichostrongylus colubriformis nematode infections in human. Emerging Infectious Diseases Journal 17, 7.
Norris, R.E., and B.R. Pearson. 1975. Fine Structure of Pyramimomas parkeae sp. nov. (Chlorophyta, Prashinophyceae). Archiv für Protistenkunde 117, 192 – 213.
Rivas, R., E. Velázquez, José Luis Zurdo-Piñeiro, Pedro F. Mateos, and E.M.N. Molina. 2004. Identification of microorganisms by PCR amplification and sequencing of a universal amplified ribosomal region present in both prokaryotes and eukaryotes. Journal of Microbiological Methods 56, 413 – 426.
Schultz, J., S. Maisel, D. Gerlach, T. Muller, and M. Wolf. 2005. A common core of secondary structure of the Internal Transcribed Spacer 2 (ITS 2) throughout the eukaryota. RNA Journal 11, 361 – 364.
Suutari, M., M. Majaneva, D.P. Fewer, B. Voirin, A. Aiello, T. Friedl, A.G. Chiarello, and J. Blomster. 1990. Molecular evidence for a diverse green algal community growing in the hair of sloths and a specific association with Trichophilus welckeri (Chlorophyta, Ulvophyceae). BioMed Central Evolutionary Biology 87, 4576 - 4579
Tamura, K., D. Peterson, N. Peterson, G. Stecher, M. Nei, and S. Kumar. 2011. MEGA5: Molecular Evolutionary Genetics Analysis using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods. Molecular Biology and Evolutionary 28, 2731 – 2739.
Woese, C.R., O. Kandler, and M.L. Wheelis. 1990. Towards a natural system of organisms: proposal for the domains Archaea, Bacteria, and Eucarya. Proceedings of the National Academy of Sciences of the USA 87, 4576 - 4579.