hrvatski

Molekularna filogenija slatkovodnih planarija (Platyhelminthes, Turbellaria, Tricladida) na osnovu mitohondrijskih gena za 16S rRNA i COI

Tricladida comprises the free-living flatworms, taxonomically grouped into four infraorders according to their ecology: Maricola, Paludicola, Cavernicola and Terricola. Our researches are based on the freshwater planarians. We choose this group of organisms because of their incomplete taxonomy and phylogeny, and relatively high number of species in such a small area. Samples were collected at many areas in Herzegovina and were stored in 96% ethanol for further researches. DNA was purified from ethanol fixed specimens using GenElute Mammalian Genomic DNA Miniprep Kit (Sigma-Aldrich, SAD) and DNA - QuickExtract DNA Extraction Solution 1.0 (Epicentre, SAD). Specific primers were used to amplify sequences for COI and 16S rRNA. The sequences were sent in the Macrogen company (Seoul, Republic of Korea) where the primary DNA structure was determined. The sequences have been analyzed, together with those from GenBank, using maximum likelihood, maximum parsimony and bayesian methods. For those analyses we used set of programs which include Mr. Bayes 3.1.1., Modeltest 3.7 and PAUP 4 beta 10. Resultes of those analyses were visualizated using Treeview 1.6.6. program. Of those two genes to be tested as markers for phylogenetic inference in the Tricladida, 16S could not be amplified despite multiple attempts under different PCR conditions. Primers used were universal and had been used previously in multiple studies. However, triclads might have sequence variations that hindered amplification. Therefor 16S was rejected. On the other side COI gen was succussfully isolated from the most of our samples and has been used for further analyses whose extend and confirm our knowledge about this issues.

planarije; mitohondrijska DNA; COI; 16s rRNA

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