Phylogenetic relationships within the genus Rana (Amphibia, Ranidae) based on a RFLP method and 16S, cytb, ITS2 sequence analysis

Korlević, Petra (2012) Phylogenetic relationships within the genus Rana (Amphibia, Ranidae) based on a RFLP method and 16S, cytb, ITS2 sequence analysis. Diploma thesis, Faculty of Science > Department of Biology.

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Abstract

Water frogs (genus Rana, subgenus Pelophylax) present an interesting research subject for ecology, physiology and genetics. Rana kl. esculenta hybridogenetic complex, which consists of two parental species (Rana ridibunda and Rana lessonae) and their hybrid species (Rana kl. esculenta), is of special interest. Hybrids often show morphological traits similar to either parent, making them harder to identify on the field. Because of that, more relevance is given to molecular and phylogenetic methods in the identification of water frog species. Identification by morphological traits has shown that in central Croatia both parental populations of R. ridibunda and R. lessonae can inhabit the same location and thus lead to the formation of hybrids. The goal of this project was to determine the phylogenetic relationships of water frog populations from Crna Mlaka based on 16S rRNA, cytochrome b and ITS2sequence analysis, and to examine the efficiency of a simple RFLP method for identifying parental species and hybrids based on the digestion of a conserved ITS2 fragment with three restriction enzymes (KpnI, HaeII and SmaI). Results of this phylogenetic analysis have shown that both parental species R. ridibunda and R. lessonae, and thus potential hybrids, inhabit Crna Mlaka. Phylogenetic analysis of mitochondrial 16S rRNA and cytb genes proved to be the best method for species and haplotype identification, while the RFLP method and phylogenetic analysis of nuclear ITS2 regions, which could help differentiate hybrid samples, need further protocol optimization.

Item Type: Thesis (Diploma thesis)
Keywords: Rana (Pelophylax), Rana kl. esculenta complex, 16S rRNA, cytb, ITS2, RFLP
Supervisor: Franjević, Damjan
Date: 2012
Number of Pages: 79
Subjects: NATURAL SCIENCES > Biology
Divisions: Faculty of Science > Department of Biology
Depositing User: Grozdana Sirotic
Date Deposited: 06 Jun 2014 07:32
Last Modified: 19 Sep 2014 11:00
URI: http://digre.pmf.unizg.hr/id/eprint/2274

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