A set of microsatellite markers for an endangered arboreal marsupial, Leadbeater's possum

Hansen, Birgita D.; Sunnucks, Paul; Blacket, Mark J.; Taylor, Andrea Carolyn

doi:10.1111/j.1471-8286.2005.01066.x

Cited by 4 publications

(5 citation statements)

References 9 publications

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Section: Methodsmentioning

confidence: 99%

“…2003). Primer sequences and polymerase chain reaction (PCR) conditions for amplification of microsatellite markers are described in Hansen et al . (2005) except for DT1, which was amplified using touchdown cycling from 62 to 55 °C.…”

Section: Methodsmentioning

confidence: 99%

“…All samples were genotyped using 14 polymorphic microsatellite markers developed for Gymnobelideus leadbeateri (GL4, GL5A, GL6, GL7, GL13, GL19B, GL24, GL28, GL33, GL35, GL38, GL39, GL42, GL44; Hansen et al 2005) and one from another petaurid, the striped possum, Dactylopsila trivirgata (DT1; Hansen et al 2003). Primer sequences and polymerase chain reaction (PCR) conditions for amplification of microsatellite markers are described in Hansen et al (2005) except for DT1, which was amplified using touchdown cycling from 62 to 55°C.…”

Section: Sample Collection and Microsatellite Genotypingmentioning

confidence: 99%

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Isolated remnant or recent introduction? Estimating the provenance of Yellingbo Leadbeater's possums by genetic analysis and bottleneck simulation

Hansen

Taylor

2008

Molecular Ecology

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Effective conservation management requires that genetically divergent populations potentially harbouring important local adaptations be identified and maintained as separate management units. In the case of the endangered Australian Leadbeater's possum (Gymnobelideus leadbeateri), an arboreal marsupial endemic to Victoria, uncertainty over the evolutionary origin of a potentially important extant wild population recently discovered in atypical habitat (lowland swamp) at Yellingbo is hampering such efforts. The population is rumoured to be a recent introduction. Microsatellite allele frequencies at Yellingbo differed substantially from those in sampled populations in montane ash forest (F(ST) between 0.23 and 0.36), and Bayesian clustering analyses of genotypes strongly separated them (K = 2). We conducted a suite of bottlenecking tests which all indicated that Yellingbo had undergone a recent reduction in size. The extent to which the distinctiveness of Yellingbo animals might be expected solely through bottlenecking associated with a recent introduction, was tested by simulating population-history scenarios seeded with genotypes from candidate wild and captive sources. No bottleneck scenario reproduced anything approaching the genetic distinction of the Yellingbo population, with all STRUCTURE analyses placing Yellingbo in a separate cluster to simulated populations (K = 2, minimum F(ST) = 0.13). These results suggest that Yellingbo does not share recent ancestry with other extant populations and instead may be a remnant of an otherwise extinct gene pool. Importantly, this may include genes involved in adaptation to a lowland swamp environment, substantially adding to the conservation importance of this population, and suggesting that separate management may be prudent until evidence suggests otherwise.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Sample Collection and Microsatellite Genotypingmentioning

confidence: 99%

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Isolated remnant or recent introduction? Estimating the provenance of Yellingbo Leadbeater's possums by genetic analysis and bottleneck simulation

Hansen

Taylor

2008

Molecular Ecology

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“…We consider the limited yield of polymorphic markers a function of the organisms rather than the techniques: good suites of markers were developed concurrently for two mammal species (e.g. Hansen et al . 2005), and the SWAPP and consensus primer protocols have been successful in our hands (e.g.…”

Section: Details Of Polymorphic Adeliini Locimentioning

confidence: 99%

Anonymous single‐copy nuclear DNA (scnDNA) markers for two endemic log‐dwelling beetles: Apasis puncticeps and Adelium calosomoides (Tenebrionidae: Lagriinae: Adeliini)

Schmuki¹,

Blacket

Sunnucks³

2006

Molecular Ecology Notes

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Three approaches — microsatellite library screening, consensus primer PCR (polymerase chain reaction) and sequencing with arbitrary primer pairs (SWAPP) — were used to develop single‐copy nuclear DNA (scnDNA) markers for log‐dwelling beetles Apasis puncticeps and Adelium calosomoides. We are unaware of other nuclear markers for Adeliini. We tested > 70 primer pairs per species, but despite exhaustive optimization, we obtained only five polymorphic markers. Nonetheless, the markers are valuable in detection of effects of habitat fragmentation.

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“…As a result, understanding the spatial distribution and identifying populations of high conservation value (from a genetic point of view) will be crucial for development of effective management measures. It has previously proven difficult to obtain polymorphic markers for the western ringtail possum and none of the primers for the brushtail possum (Spencer and Bryant 2001, Taylor and Cooper 1998Hansen et al 2005, Mitrovski et al 2005 were found to amplify in the ringtail possum. As such it was necessary to find a species-specific alternative.…”

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confidence: 98%

Isolation and characterisation of polymorphic microsatellite markers in the western ringtail possum, Pseudocheirus occidentalis

Wilson

Tores²,

Spencer

2009

Conservation Genet Resour

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We report on the development of 14 novel polymorphic microsatellite markers from the western ringtail possum (Pseudocheirus occidentalis), a ''threatened'' (Vulnerable) arboreal marsupial, endemic to the biodiversity hotspot region of south-west of Western Australia. The markers were developed to obtain population genetic data to quantify any changes which may be associated with increasing geographical fragmentation. We describe the diversity among 66 unrelated (adult) P. occidentalis individuals. Overall, the markers produced between three to eight alleles per locus and observed and expected heterozygosities of 59% and 69%, respectively. Due to predation by introduced foxes, urban encroachment changes in fire regime and the resultant levels of fragmentation in its range, these microsatellite markers are an important tool for evaluating population genetic information and for prioritising populations for conservation management.The western ringtail possum (Pseudocheirus occidentalis) is a medium sized nocturnal marsupial weighing up to 1.3 kg. It is endemic to south-western Australia (Van Dyck and Strahan 2008). The species is under increasing threat from introduced (cats and foxes) and native (pythons and chuditch) predators, changes in fire management practices and habitat fragmentation (Van Dyck and Strahan 2008). It has suffered localised decline over much of its range and localised extinction from several inland locations. As a result, understanding the spatial distribution and identifying populations of high conservation value (from a genetic point of view) will be crucial for development of effective management measures. It has previously proven difficult to obtain polymorphic markers for the western ringtail possum and none of the primers for the brushtail possum (Spencer and Bryant 2001, Taylor and Cooper 1998Hansen et al. 2005, Mitrovski et al. 2005 were found to amplify in the ringtail possum. As such it was necessary to find a species-specific alternative. Here we report on the development of polymorphic microsatellite loci which provide sufficient resolution for detecting fine-scale population genetic structure, gene flow and measures of population genetic diversity.Ninety-eight plasmid clones were sequenced (Genebank accession numbers GQ222585-GQ222683) and 30 microsatellite-containing regions were selected based on a minimum of 200 base pairs (bp) of high sequence-quality flanking either side of the repetitive element to design primers. Primer Express (Version 2.0; Applied Biosystems) was used to design a set of primers for each of the 30 sequences, using the following parameters: (i) Tm between 57°C and 63°C, (optimal 59°C); (ii) GC content of 45-55%; a 3 0 GC clamp of 3 residues; (iii) primer length 20-26 bp (optimum 24); and (iv) the PCR amplicon size of 80-350 bp. Primer pairs for these 30 sequences were ordered with an M13 tag (Schuelke 2000) and optimised to work in a 30 ll volume temperature-gradient PCR (55-65°C) containing 19 reaction buffer, 2.5 mM MgCl 2 , 0.1 mM bovine serum

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A set of microsatellite markers for an endangered arboreal marsupial, Leadbeater's possum

Cited by 4 publications

References 9 publications

Isolated remnant or recent introduction? Estimating the provenance of Yellingbo Leadbeater's possums by genetic analysis and bottleneck simulation

Isolated remnant or recent introduction? Estimating the provenance of Yellingbo Leadbeater's possums by genetic analysis and bottleneck simulation

Anonymous single‐copy nuclear DNA (scnDNA) markers for two endemic log‐dwelling beetles: Apasis puncticeps and Adelium calosomoides (Tenebrionidae: Lagriinae: Adeliini)

Isolation and characterisation of polymorphic microsatellite markers in the western ringtail possum, Pseudocheirus occidentalis

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