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Zhang, F., Zhao, S., Kan, X., Chen, B., & Ling, J. , Phylogenetic relationship among geographic subspecies of cistoclemmys flavomarginata in eastern china - abstract. Unpublished paper presented at Program and Abstracts of the Tenth Annual Symposium on the Conservation and Biology of Tortoises and Freshwater Turtles. 
Added by: Sarina Wunderlich (06 Jul 2014 16:17:31 UTC)
Resource type: Conference Paper
BibTeX citation key: Zhang2013
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Categories: General
Keywords: Genetik - genetics, Habitat - habitat, Schildkröten - turtles + tortoises, Südostasien - South East Asia, Testudinidae, Testudo graeca
Creators: Chen, Kan, Ling, Zhang, Zhao
Collection: Program and Abstracts of the Tenth Annual Symposium on the Conservation and Biology of Tortoises and Freshwater Turtles
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Views index: 41%
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Cistoclemmys flavomarginata, or Yellow-margined Box Turtle, is an Endangered species listed by International Union for Conservation of Nature (IUCN). It is mainly distributed in Southeastern China, Twain and Japan. It is now very rare to find wild C. flavomarginata in natural habitats, possibly owing to the deterioration of environment. On the other hand, C. flavomarginata has been found to be effective in preventative or therapeutic medicine, such as showing high efficacy as a cancer adjuvant therapy. This situation further threatens the protection of C. flavomarginata. Considering the current research more focused on the morphology, anatomy, breeding and farming, it is imperative to conduct genetic studies to clarify variations among the subspecies of C. flavomarginata, thus providing us the guideline for distribution and conservation. By utilizing PCR coupled with DNA sequencing approach, we have analyzed 83% of mitochondria genome sequences of four pieces of wild C. flavomarginata collected from three different geographic locations. With the use of high fidelity DNA polymerase and double validation of DNA sequences by software and manual inspection, a complete alignment was performed among these four C. flavomarginata mtDNAs along with the published Cuora flavomarginata mtDNA sequence as the control. It was identified that most genes (e.g., 16S rRNA, tRNAs, ND4, ND 5) have 1–5 sites mutated, which is not sufficient to perform genetic linkage analysis. However, cytochrome b (Cytb), a well studied and reliable genetic marker, displays 10 mutations along its full length sequence. A phylogenetic tree was then built using ClustalW2/EBI software. It was revealed that the evolutionary distance among these C. flavomarginata is well correlated with their geographic separation. Interestingly, one specimen of C. flavomarginata from the same area with previously studied Cuora flavomarginata showed the highest conservation in their Cytb sequences. Being the most suitable marker to identify taxonomic relationship at subspecies level and to resolve relatively recent evolutionary history, Cytb mutations normally occur at the silent positions. However, our data showed one amino acid (Aa) mutation (Ile to Thr or Ala to Thr) among three pieces of C. flavomarginata; no Aa sequence variation was found in the one closest to the control. This result suggests a potential perturbation in Cytb biochemical function that links to physiological behavior of geoemydidae. In summary, our study here defined an accurate genetic marker with Cytb to identify wild geo-subspecies of C. flavomarginata. With further validation using larger populations, we anticipate developing a robust diagnosis method to provide practical guidance for the evolutionary and conservative studies of C. flavomarginata. Future research will be focused on the genome analysis of C. flavomarginata and related geoemydids.
Added by: Sarina Wunderlich  
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