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Genetic diversity and distinctiveness of Plectrohyla guatemalensis (Anura: Hylidae) in Guatemala

Zamora Jerez, Olga

[Thesis]. Manchester, UK: The University of Manchester; 2014.

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Abstract

Microsatellite markers are acquiring more attention for the study and conservation of populations. They can help scientists understand and ask a variety of questions about populations. Next Generation Sequencing technologies (NGS) are becoming the preferred method for scientists to develop microsatellites since they can produce large amounts of data in less time than conventional cloning. P. guatemalensis is a hylid frog that inhabits forests of Guatemala, México and Honduras. In this project microsatellite markers were developed using both methods to do a first assessment on the genetic status of P. guatemalensis in Guatemala.Microsatellites primers were developed using both conventional cloning techniques at Sheffield University (SHF) and NGS. Twenty one pairs of primers were developed using data generated from an Illumina Miseq sequencer while 16 primers were developed using cloning methods in Sheffield. In total a set of eight polymorphic loci were used to evaluate the population genetics of P. guatemalensis.This study shows the use of NGS technologies and bioinformatics tools to be a quick and effective way of obtaining Possible Amplifiable Loci (PALs). In this project only a few loci could be tested and more experiments are needed before being able to determine if one method is advantageous over the other. Results showed that the observed heterozygosity was higher than expected for seventeen loci across all populations, and divergence from Hardy Weinberg Equilibrium was found in seven loci in Laj Chimel and Tatasiriré populations and four loci from Trifinio. The inbreeding coefficient (FIS) varied from -0.578 to 0.633 across populations. The negative value for some loci suggests possible outbreeding, however, other loci show a strong positive value for Fis potentially indicating small scale substructure within populations. FST showed clear population differentiation between the three localities, with a value of 0.302. The localities were P. guatemalensis was found are in danger of disappearing entirely due to habitat loss, pollution and emergent infectious diseases. It is important to do constant surveys in order to plan properly the conservation of amphibian species in these threatened forests. The microsatellites developed during this study will be useful for future of conservation genetics programs of amphibians in Guatemala.