Development and optimisation of molecular assays for microsatellite genotyping and molecular sexing of non-invasive samples of the ghost bat, Macroderma gigas
| Autor: | Jamie N. Tedeschi, Margaret Byrne, Peter S. Spencer, Kym Ottewell, Rujiporn Thavornkanlapachai, Kyle N. Armstrong, Brad Durrant, Chris Knuckey, Shelley McArthur |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Male Genotype Genotyping Techniques Population Population genetics Sexing 03 medical and health sciences Feces 0302 clinical medicine Chiroptera Genetics Animals education Molecular Biology Genotyping Alleles education.field_of_study Genetic diversity Polymorphism Genetic biology Australia Macroderma gigas Genetic Variation High-Throughput Nucleotide Sequencing General Medicine biology.organism_classification 030104 developmental biology Genetics Population Evolutionary biology 030220 oncology & carcinogenesis Microsatellite Female Genetic monitoring Microsatellite Repeats |
| Zdroj: | Molecular biology reports. 47(7) |
| ISSN: | 1573-4978 |
| Popis: | The ghost bat (Macroderma gigas) is endemic to Australia but is under threat, with scarce information available on the genetic health of remaining populations. Here, we develop molecular assays for microsatellite genotyping and molecular sexing of non-invasive samples as a genetic monitoring tool to identify individuals, measure genetic diversity and investigate spatial and temporal patterns of habitat use by ghost bats. We identified novel microsatellites through high-throughput sequencing on the Illumina MiSeq platform. Of 48 loci tested, six markers were added to five previously developed microsatellite loci. We developed three Y-linked (DDX3Y, Zfy and SRY) and one X-linked markers (Zfx) to enable molecular identification of sex. To assess performance, all 11 microsatellite and four sex-linked markers were amplified in three multiplex reactions in 160 M. gigas faecal samples from the Pilbara region, Western Australia. The combined markers offered a high level of individual discrimination (PIDsibs = 0.00002) and we detected 19 bats in total (11 males, 4 females and 4 sex undetermined). The number of alleles per locus ranged from 5 to 14 and the average observed and expected heterozygosity across loci were Ho = 0.735 (0.58–0.91) and uHe = 0.785 (0.59–0.89) respectively. Our molecular assays allowed identification of individuals from faecal samples at multiple time points and spatial locations and enabled us to elucidate patterns of habitat usage at the study site. This study highlights the value of our molecular assays as a potential capture-mark-recapture technique for population monitoring for this species. |
| Databáze: | OpenAIRE |
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