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DNA sampling of Little Penguin (Eudyptula minor) colonies in south-eastern Australia Rebecca Overeem1, Amanda Peucker1, Peter Dann2 & Chris Burridge1 3 1Deakin University, Warrnambool Campus, Victoria, Australia Island Nature Parks, Cowes, Victoria, Australia 3Otago University, Dunedin, New Zealand 2Phillip Cartoon: Vic Nelson Little Penguin (Eudyptula minor) • World’s smallest penguin • 33cm in height • Weight ~ 1kg • First breed at 2-3 years • Average lifespan of 6-7 years VICTORIA Melbourne • • • • Endemic to southern Australia and New Zealand Distributed from Fremantle in Western Australia to northern New South Wales and Tasmania 40 000 breeding pairs in Victoria Phillip and Gabo Islands contain the largest colonies Conservation Priority • Human induced pressure • Potentially limited movement between colonies • Phenotypic variation: potential that individual colonies are locally adapted Aim • Determine exchange of reproductive individuals among colonies – H0: Little Penguin colonies exist in a meta-population structure Southern Ocean Melbourne Approach • Survey variation of: – microsatellite loci – mtDNA Methods • Seven sites – 50 adults per site Granite Island St Kilda Gabo Island London Bridge Phillip Island Kangaroo Island Middle Island Southern Ocean Molecular Methods Microsatellites – Nuclear DNA – Repeating units of 1-6 bp – Distributed throughout the genome • 17 loci screened for polymorphism – Roeder et al. 2002 Adelie Penguin (4 loci) – Akst et al. 2002 Galapagos & Magellanics (6 loci) – Schlosser et al. 2003 Humboldt Penguin (7 loci) Molecular Methods continued Mitochondrial DNA – Genetic information contained in the mitochondria – Maternally inherited – Rapid rate of evolution 650bp segment of the control region sequenced, employing primers of Roeder et al. 2002 6 variable nucleotides identified, 3 PCR-RFLPs developed Results Microsatellites • 17 loci screened – 5 polymorphic • Akst et al. 2002 (B3-2, G3-11, G2-2) • Roeder et al. 2002 (AM13) • Schlosser et al. 2003 (Sh1Ca9) • All in agreement with Hardy-Weinberg equilibrium except G2-2 at Kangaroo Island • Genotypes independent across loci • Only Kangaroo Island distinguished from some other colonies • No relationship between genetic divergence and geographic distance 0.40 0.35 0.30 0.25 FST 0.20 Results Mitochondrial DNA 0.15 0.10 0.05 0.00 -0.05 0 200 400 600 800 1000 1200 1400 Distance (km) • PCR-RFLPs screened • All colonies polymorphic • Kangaroo and Granite Islands distinct from the Victorian colonies • Almost significant relationship between genetic divergence and geographic distance (Mantel test P-value = 0.054) 1600 Pairwise population comparisons Southern Ocean Further Study • Extend microsatellite and mtDNA research to incorporate – a broader geographic range (TAS, NSW, WA, NZ) – more microsatellite loci Acknowledgements • Staff and students at Deakin University, Warrnambool campus – Colleagues in the Molecular Ecology and Biodiversity Laboratory • Staff at Phillip Island Nature Park • This study was partially funded by: - Deakin University (RLO-Scholarship) - Holsworth Wildlife Research Fund (RLO) - Norman Wettenhall Foundation (RLO) - Australian Research Council (AJP-Scholarship) - Phillip Island Nature Parks (AJP) • Field assistants