The pioneering use of ISSR (Inter Simple Sequence Repeat) in Neotropical anurans: preliminary assessment of genetic diversity in populations of Physalaemus cuvieri (Amphibia, Leiuperidae)

Abstract

The greatest diversity of anurans in the world is in Brazil and one of the major challenges is to reconcile the accelerated economic development with strategies that aim to maintain this diversity in forest fragments, often representing ESUs of some biomes. This study aimed to obtain data that will support conservation projects through the pioneering use of ISSR analysis in Neotropical anurans, estimating the intra-and interpopulation genetic diversity of four populations of P. cuvieri (Paraná and São Paulo regions). Of the 65 loci scored 58 were polymorphic, with 0.797 intrapopulation variation and 0.203 interpopulation variation. The index of interpopulation genetic diff erentiation (F ST ) proved to be high among the population of Marmeleiro-PR and the three populations of SP (F ST > 0.288); genetic dissimilarity was related to the geographical distance. The ISSR proved to be effi cient and useful molecular markers in comparison with other markers most widely used for preliminary diagnosis of genetic diversity in populations of amphibians, and could be applied as a tool for future conservation projects, since they could identify potential ESUs and infl uence decisions on the preservation of fragments.

Genetic diversity of Arapaima gigas (Schinz, 1822) (Osteoglossiformes: Arapaimidae) in the Araguaia-Tocantins basin estimated by ISSR marker

“…Due to its ease of use, low cost and high sensitivity, ISSR markers have been employed in studies on genetic diversity of several species of plants and animals (Luque et al, 2002;Bornet & Branchard, 2004;Askari et al, 2011;Moresco et al, 2013), as well as studies of sexual differentiation in plants (Ehsanpour et al, 2008) and identification of hybrid fish (Bignotto et al, 2009;AlmeidaFerreira et al, 2011). This marker has allowed the analysis of the main parameters used for determining the genetic diversity and the level of differentiation between species and natural populations of Neotropical fish (amount of polymorphic loci, expected or average heterozygosity and the number of migrants per generation) (Paiva et al, 2006;Sofia et al, 2006;Lopes et al, 2008;Antunes et al, 2010;Almeida-Ferreira et al, 2011;Domingos et al, 2014).…”

“…The possibility of estimating the genetic variability by polymorphisms in the DNA of an organism, as well as of the popularization of molecular techniques, has encouraged progress in studies of population genetics (Antunes et al, 2010;Moresco et al, 2013). These molecular markers are important tools for estimating parameters, such as levels of genetic diversity within populations and magnitudes of gene flow between populations (Avise, 1996;Woodruff, 2001).…”

Molecular identification of Todiramphus chloris subspecies on the Arabian Peninsula using three mitochondrial barcoding genes and ISSR markers

“…Here, 90% of the primers were based on di-nucleotides (Table 1). Several reports stated that di-nucleotide repeats are more agreeable to ISSR investigation than other nucleotide repeats (Askari et al, 2011, Moresco et al, 2013). The selected ISSR primers gave 118 bp amplified bands (Fig.…”

Incorporating Molecular Genetics into Remote Expedition Fieldwork

“…ISSRs are DNA fragments produced by single primer PCRs. The primer is an 18–20 bp anchored microsatellite that is useful on a suite of species [27, 28] and was used to document a positive DNA extraction. Qiagen PCR reaction volume was 25 μL and included Illustra PuReTaq Ready-To-Go PCR Beads, 22.0 μL dH20, 2.0 μL primer and 1.0 μL DNA.…”