Characterization, multiplex conditions, and cross‐species utility of tetranucleotide microsatellite loci for Blanchard's cricket frog (<i>Acris crepitans blanchardi</i>)

Beauclerc, Kaela B.; Johnson, Bob; White, Bradley N.

doi:10.1111/j.1471-8286.2007.01874.x

Cited by 4 publications

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References 3 publications

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“…Tetranucleotide microsatellites were isolated according to the protocol of Beauclerc et al (2007). In total, 115 clones were sequenced, of which 83 were positive for a tetranucleotide repeat, with five duplicate clones found.…”

Section: Microsatellite Development and Profilingmentioning

confidence: 99%

Genetic rescue of an inbred captive population of the critically endangered Puerto Rican crested toad (Peltophryne lemur) by mixing lineages

2009

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The Puerto Rican crested toad (Peltophryne lemur) is currently composed of a single wild population on the south coast of Puerto Rico and two captive populations founded by animals from the northern and southern coasts. The main factors contributing to its decline are habitat loss, inundation of breeding ponds during storms, and impacts of invasive species. Recovery efforts have been extensive, involving captive breeding and reintroductions, habitat restoration, construction of breeding ponds, and public education. To guide future conservation efforts, genetic variation and differentiation were assessed for the two captive colonies and the remaining wild population using the mitochondrial control region and six novel microsatellite loci. Only two moderately divergent mitochondrial haplotypes were found, with one fixed in each of the southern and northern lineages. Moderate genetic variation exists for microsatellite loci in all three groups. The captive southern population has not diverged substantially from the wild population at microsatellite loci (F ST = 0.03), whereas there is little allelic overlap between the northern and southern lineages at five of six loci (F ST [ 0.3). Despite this differentiation, they are no more divergent than many populations of other amphibian species. As the northern breeding colony may not remain viable due to its small size and inbred nature, it is recommended that a third breeding colony be established in which northern and southern individuals are combined. This will preserve any northern adaptive traits that may exist, and provide animals for release in the event that the pure northern lineage becomes extirpated.

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Section: Microsatellite Development and Profilingmentioning

confidence: 99%

Genetic rescue of an inbred captive population of the critically endangered Puerto Rican crested toad (Peltophryne lemur) by mixing lineages

2009

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“…We amplified the DNA template at four microsatellite markers (Acr-3, Acr-28, Acr-2, and Acr-14) fluorescently labeled with Applied Biosystems DS-33 set (NED, 6-FAM, 6-FAM, and VIC dyes, respectively), following the program outlined by Beauclerc et al . [ 64 ], via polymerase chain reaction (PCR). We ran all four primers together in one multiplex reaction, using Type-It Microsatellite PCR Kits (Qiagen, Germantown, Maryland, USA), following the manual for reagent concentrations and volumes of reagents (Type-it Microsatellite PCR Handbook, October 2010).…”

Section: Methodsmentioning

confidence: 99%

“…We considered amplification success for a sample at a given microsatellite marker binary, determined by whether PCR allowed for allele scoring, with clear homozygote or heterozygote peaks visible and near or within the expected base pair range, as per Beauclerc et al . [ 64 ]. We calculated the proportion of successful amplifications for each sample type.…”

Section: Methodsmentioning

confidence: 99%

Comparing skin swabs, buccal swabs, and toe clips for amphibian genetic sampling, a case study with a small anuran (Acris blanchardi)

Rainey,

Tryc,

Nicholson

2024

Biology Methods and Protocols

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Multiple methods for collecting genetic samples from amphibians exist, each with their own implications for study design, animal welfare, and costs. Toe clipping is one common method, but there is ongoing debate regarding its potential detriment. Less invasive methods should be implemented, if efficacious, as amphibians are a particularly vulnerable vertebrate group. Skin and buccal swabbing are less invasive methods for genetic sampling, but the potential for contamination and a lower yield of DNA may exist. To compare these methods, we gathered skin swabs, buccal swabs, and toe clips from the same individuals of a relatively small anuran species, Blanchard’s Cricket Frog (Acris blanchardi). We then compared DNA yield, DNA purity, amplification success rate, and genotypic data quality among sample types. We found toe clips and buccal swabs generated similar DNA yield and purity, with skin swabs yielding significantly less DNA of significantly lower purity than the other sample types. Amplification success rate was significantly higher using toe clips compared to the other sample types, though buccal swab samples amplified more readily than skin swabs. Genotypic data from toe clips and buccal swabs did not differ significantly in quality, but skin swab data quality was significantly lowest among sample types. Thus, skin swabbing could produce erroneous data in some situations, but buccal swabbing is likely an effective substitute to toe clipping, even for small species. Our results can help future researchers select which genetic sampling method might best suit their research needs.

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“…For example, amphibian populations have considerable natural variation in sensitivity to environmental contaminants (Bridges et al 2001) and some southern populations of cricket frogs apparently have developed resistance to the lethal pesticide DDT (dichlorodiphenyltrichloroethane) (Boyd et al 1963). However, the reduced genetic diversity for both mtDNA and nuclear microsatellite markers (Beauclerc et al 2007) in northern populations of Blanchard's Cricket Frog could prevent them from adapting to such damaged habitats in the future (Frankham et al 2002).…”

Section: Application Of Genetic Data To Conservation Recommendationsmentioning

confidence: 98%

Distinctiveness of declining northern populations of Blanchard’s Cricket Frog (Acris blanchardi) justifies recovery efforts

2010

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Peripheral populations of widespread species are often considered unworthy of conservation efforts; however, they may be adapted to the conditions found at the range edge and are therefore important to the future evolutionary potential of the species. Blanchard's Cricket Frog (Acris blanchardi Harper, 1947) is widespread and abundant throughout the central United States, but is declining at the northern edge of its range. To assess the distinctiveness and conservation value of the northern populations, we investigated the spatial genetic structure and phylogeography of this anuran using mitochondrial control region sequences. Analysis of 479 individuals identified 101 haplotypes, with relatively low nucleotide diversity. Two moderately divergent clades were found. One was restricted to the southwest, which was probably a refugium during the Pleistocene, whereas the other occurred primarily across the north and is likely the result of postglacial colonization. The genetic distinctiveness of northern populations indicates the potential for adaptive differences of individuals in this region relative to those in the south. We therefore conclude that conservation efforts are justified for the declining northern populations of Blanchard's Cricket Frog, and we use the spatial genetic structure described here to develop specific recommendations for this anuran.Résumé : On considère souvent que les populations périphériques d'espèces à large répartition ne valent pas les efforts de conservation; elles peuvent, cependant, être adaptées aux conditions qui prévalent à la périphérie de l'aire de répartition et ainsi être importantes pour le potentiel évolutif futur de l'espèce. La rainette grillon de Blanchard (Acris blanchardi Harper, 1947) est une espèce largement répandue et abondante dans tout le centre des É tats-Unis, mais elle est en déclin à la marge nord de son aire de répartition. Afin d'évaluer la particularité et la valeur de conservation des populations nordiques, nous avons étudié la structure génétique et la phylogéographie de cet anoure à l'examen des séquences de la région de contrôle mitochondriale. Une analyse de 479 individus a révélé l'existence de 101 haplotypes, avec une diversité relativement basse des nucléotides. Il existe deux clades à divergence moyenne. L'un est restreint à la région sud-ouest, qui a probablement servi de refuge au pléistocène, alors que l'autre est surtout répandu dans tout le nord et résulte vraisemblablement de la colonisation postglaciaire. La différenciation génétique des populations nordiques laisse croire à un potentiel de différences adaptives chez les animaux de cette région par rapport aux animaux du sud. Nous concluons qu'il est justifié de déployer des efforts de conservation pour les populations nordiques déclinantes de la rainette grillon de Blanchard; nous utilisons la structure génétique spatiale décrite ici pour mettre de l'avant des recommandations spécifiques au sujet de cet anoure.[Traduit par la Rédaction]

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Characterization, multiplex conditions, and cross‐species utility of tetranucleotide microsatellite loci for Blanchard's cricket frog (Acris crepitans blanchardi)

Cited by 4 publications

References 3 publications

Genetic rescue of an inbred captive population of the critically endangered Puerto Rican crested toad (Peltophryne lemur) by mixing lineages

Genetic rescue of an inbred captive population of the critically endangered Puerto Rican crested toad (Peltophryne lemur) by mixing lineages

Comparing skin swabs, buccal swabs, and toe clips for amphibian genetic sampling, a case study with a small anuran (Acris blanchardi)

Distinctiveness of declining northern populations of Blanchard’s Cricket Frog (Acris blanchardi) justifies recovery efforts

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