Nest-site selection and the factors influencing hatching success and offspring phenotype in a nocturnal skink. Supplementary Figures

<p>Nest-site selection in ectothermic animals influences hatching success and offspring phenotype, and it is predicted that females should choose nesting sites that maximise their reproductive fitness, ultimately through the reproductive success of their offspring. We completed nest-site choice experiments on a nocturnal lizard, the egg-laying skink (<i>Oligosoma suteri</i>), to determine whether eggs (and subsequent hatchlings) from cooler nests do better at cooler incubation temperatures, and conversely if those laid in warmer nests perform better at warmer incubation temperatures. We provided a simple nest-choice experiment, with oviposition-retreat sites available in either a hot or a cool sector of the enclosure; in the wild females nest under objects. Female <i>O. suteri </i>laid eggs both during the day and night, and nested more in the hot than cool sector. Eggs from each clutch were split across three egg incubation temperatures (18°C, 22°C, 26°C) to decouple the impact of initial nest-site choice from the subsequent incubation temperature regime. Whether eggs were initially laid in the hot or cool sector was not related to hatching success, offspring phenotype or offspring locomotor performance. We conclude that offspring phenotype and performance is primarily influenced by the temperature during incubation, rather than the initial thermal environment of the nest location. Thus, female <i>O. suteri </i>may select warmer nesting sites to ensure higher incubation temperature and enhanced offspring fitness.</p

Apteryx owenii_microsatellite_genotypes_April 2013

Apteryx owenii_microsatellite_genotypes_April 201

Oligonucleotide primers used in this study.

<p>The letters L and H refer to the light and heavy strands. Values in ‘5’ position' refer to the position of the 5′ position in the complete <i>Eumeces egregius</i> mtDNA sequence <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034599#pone.0034599-Kumazawa1" target="_blank">[96]</a>.</p

Mean TrN genetic distances between <i>control region</i> haplotypes.

<p>Mean TrN genetic distances between <i>control region</i> haplotypes.</p

<i>Control region</i> haplotype network for the Otago skink.

<p>Each circle represents one haplotype and the size indicates the number of individuals with each haplotype. The lines indicate one base pair change between sequences. The different colours indicate the population(s) in which each haplotype is found. A) Wild Otago skink populations, B) Wild and captive Otago skinks, C) Wild Otago skinks and introgressed scree skinks.</p

Estimates of genetic diversity (mitochondrial <i>control region</i>) within Otago skink populations and regions.

<p><i>n</i> = sample size, <i>h</i> = number of haplotypes (the specific haplotypes present are indicated), <i>Hd</i> = haplotypic diversity, <i>M</i> = total number of mutations, <i>S</i> = number of segregating (polymorphic) sites, π = nucleotide diversity, RI = raggedness index, SSD = sum of squared deviations. Asterisks indicate significant Tajima's <i>D</i>, Fu's <i>F</i>s statistic, RI and SSD values.</p

Distribution of the Otago skink and several closely related species of New Zealand skinks.

<p>The distribution of <i>O. otagense</i> (black circles), <i>O. waimatense</i> (blue squares), <i>O. acrinasum</i> (green circles), <i>O. infrapunctatum</i> (red triangles), <i>O. taumakae</i> (orange square), and <i>O. pikitanga</i> (purple circle) is indicated. Several localities mentioned in the text are highlighted, including the locations of the <i>O. otagense</i> and <i>O. waimatense</i> samples used in this study. The dashed line indicates the estimated former distribution of <i>O. otagense </i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034599#pone.0034599-Whitaker1" target="_blank">[26]</a>. The distribution data is adapted from the Department of Conservation's BioWeb Herpetofauna database. Inset: Major geographic regions of New Zealand.</p

Locality information and GenBank accession numbers for samples used in the phylogenetic analyses.

<p>Samples with CD or FT codes were obtained from the National Frozen Tissue Collection (NFTC) housed at Victoria University of Wellington, New Zealand. Samples with RE codes were obtained from ethanol preserved specimens housed at Te Papa, National Museum of New Zealand, Wellington (S codes refer to specimens from the former Ecology Division collection, now housed at Te Papa).</p

Mean model-corrected genetic distances (<i>ND2</i>, <i>ND4</i>, Cytochrome b) between the Otago skink and several closely-related species.

<p>Mean model-corrected genetic distances (<i>ND2</i>, <i>ND4</i>, Cytochrome b) between the Otago skink and several closely-related species.</p