Population genetics of east Antarctic sea urchins

Population connectivity and gene flow in near shore Antarctic Echinoids (Sterechinus neumayeri, Abatus nimrodi and Abatus ingens) was investigated in East Antarctica. This data set consists of microsatellite genotype data from 11 novel loci and mitochondrial DNA sequences from two gene region, COI and 16S. In addition, to determine if changes in temperature and salinity impacted fertilisation success in S. neumayeri, and to determine the appropriate sperm to egg ratio for this type of experiment, a fertilisation experiment was completed using various combinations of temperature, salinity and sperm to egg ratio. Samples were collected near two Australian Antarctic research stations, Casey and Davis, during the 08/09 and 09/10 summer field seasons.To generate the microsatellite data set, a total of 545 adults, nuemayeri and 26 echinoplutei were collected. Spatial replication was achieved by comparing adult populations between two regions (Casey and Davis). These two regions are separated by approximately 1400 km. Sampling in the Casey region was done at two locations 9 km apart and in the Davis region at five locations separated by 5 - 30 km. Within each location 25-50 individuals were collected from up to three sites approximately 0.5 km apart. Within each site, all individuals were collected within an area less than 50 m2. Adult urchins were collected by dip nets, snorkel or scuba depending on location. Echinoplutei were collected from the water column in two locations in the Davis region using a purpose built plankton net. DNA was extracted using QiagenDNeasy Blood and Tissue extraction kits as per the manufacturer's protocols. PCR amplification was carried out in four multiplex reactions and analysis of the PCR product was carried out on a CEQ 8000 (Beckman Coulter) automated sequencer by capillary separation, and alleles scored as fragment size using CEQ 8000 Genetic Analysis System software (ver. 8.0).Data available: Data consists of 571 individual genotypes at 11 loci in an excel spreadsheet following the GenAlEx v 6.41 layout. Sites from the Davis region are; Old Wallow 1 (OW1), Old Wallow 2 (OW2), Boyd Island (BO1), Ellis Fjord 1 (EL1), Ellis Fjord 2 (EL2), Ellis Fjord 3 (EL3), Trigwell Island 1 (TR1), Trigwell Island 2 (TR2), Trigwell Island 3 (TR3), Zappit Point 1 (ZP1), Zappit Point 2 (ZP2), Zappit Point 3 (ZP3). Sites from the Casey region are; Browning Peninsula 1 (CB1), Browning Peninsula 2 (CB2), Browning Peninsula 3 (CB3), Sparkes Bay 1 (CS1), Sparkes Bay 2 (CS2).Echinoplutei samples are Hawker Island (D1); Kazak Island 1 (K1); Kazak Island 2 (K2) Data is coded as fragment length, with a zero value representing no data.To generate the mtDNA sequence data, a total of 24 S. neumayeri individuals were sequenced for the COI gene region with two haplotypes found. For the 16S gene region, 25 individuals were sequenced with three haplotypes founds. For Abatusingens, 51 individuals were sequenced with six CO1 haplotypes and five 16S haplotypes. For Abatus nimrodi (n = 48) there were two CO1 haplotypes and eight 16S haplotypes. In addition, eight A. shackeltoni, four A. philippii and one A. cavernosus sample were included from the Davis region.Data available: data are available in four FASTA text format files, one for Abatus COI data, one forAbatus 16S data, one for Sterechinus COI data. Individuals are coded with the first two letters representing species (SN = S. neumayeri, AN = A. nimrodi, AI = A. ingens, AS = A. shackletoni, AC= A. cavernosus) the next two representing gene region (CO = COI, 16 = 16S) and either three or four more digits for Davis region samples or five digits beginning with 41 for Casey region samples.To generate the fertilisation data set, S. neumayeri were collected from Ellis Fjord prior to ice breakout. A total of 12 individuals were screened for the fertilisation experiment, seven males and five females to ensure a suitable cross where greater than 90% fertilisation success was achievable. Sperm were activated with FSW at -1.8 degrees C and sperm concentration determined using a haemocytometer. Three temperature treatments, (-1.8 degrees C, 1 degrees C and 3 degrees C), three salinity treatments (35ppt, 30ppt and 25ppt), and five sperm to egg ratios (50:1, 100:1, 500:1, 1500:1 and 2500:1) were used during fertilisation, with four replicates at each temperature:salinity:sperm to egg ratio combination. After 30 min, three to five drops of 10% formalin were added to each vial to fix eggs and to prevent further fertilisation from occurring. To determine percentage fertilisation, the first 100 eggs encountered from each vial were scored as either fertilised or unfertilised based on the presence or absence of an elevated fertilisation membrane.Data available: Data are available as an excel file, with three spreadsheets, one for each temperature treatment. Each spreadsheet consists of three tables, one for each salinity treatment. Each salinity treatment table consists of five columns. From left to right these are; sperm : egg ratio - Sperm to egg ratio, rep. No. - replicate number,        Fert. - number of fertilised eggs countedUnfert.    - number of unfertilised eggs countedMean- mean number of fertilised eggs counted

本研究调查了南极近岸海胆（斯特雷奇努斯·纽迈耶海胆Sterechinus neumayeri、尼姆罗德阿巴特斯海胆Abatus nimrodi及英根斯阿巴特斯海胆Abatus ingens）在东南极洲的种群连通性与基因流。该数据集包含来自11个新位点的微卫星基因型数据（microsatellite genotype data）及两个基因区域（COI与16S）的线粒体DNA（mitochondrial DNA）序列。此外，为探究温度与盐度变化对纽迈耶海胆S. neumayeri受精成功率的影响，并确定此类实验的适宜精卵比，研究人员采用温度、盐度及精卵比的多种组合完成了受精实验。

样本采集于澳大利亚两座南极科考站（凯西站Casey与戴维斯站Davis）附近，时间为08/09及09/10夏季科考季。

微卫星数据集生成：共采集545只纽迈耶海胆成体及26只海胆幼体（echinoplutei）。通过比较凯西与戴维斯两区域的成体种群实现空间重复（两区域相距约1400公里）。凯西区域在相距9公里的两个地点采样，戴维斯区域在相距5-30公里的五个地点采样；每个地点从至多三个相距约0.5公里的站点采集25-50个个体，每个站点内的个体采集范围均小于50平方米。成体海胆根据地点不同采用抄网、浮潜或水肺潜水方式采集；海胆幼体则通过定制浮游生物网从戴维斯区域的两个地点水体中采集。

微卫星数据可用信息：数据以遵循GenAlEx v6.41格式的Excel表格呈现，包含571个个体在11个位点的基因型。戴维斯区域站点包括：Old Wallow 1（OW1）、Old Wallow 2（OW2）、Boyd Island（BO1）、Ellis Fjord 1（EL1）、Ellis Fjord 2（EL2）、Ellis Fjord 3（EL3）、Trigwell Island 1（TR1）、Trigwell Island 2（TR2）、Trigwell Island 3（TR3）、Zappit Point 1（ZP1）、Zappit Point 2（ZP2）、Zappit Point 3（ZP3）；凯西区域站点包括：Browning Peninsula 1（CB1）、Browning Peninsula 2（CB2）、Browning Peninsula 3（CB3）、Sparkes Bay 1（CS1）、Sparkes Bay 2（CS2）；海胆幼体样本来自Hawker Island（D1）、Kazak Island 1（K1）、Kazak Island 2（K2）。数据以片段长度编码，零值代表无数据。

线粒体DNA序列数据集生成：共对24只纽迈耶海胆S. neumayeri的COI基因区域测序，发现2种单倍型（haplotype）；对25只个体的16S基因区域测序，发现3种单倍型。英根斯阿巴特斯海胆Abatus ingens的51个个体测序结果显示：CO1基因区域有6种单倍型，16S基因区域有5种单倍型；尼姆罗德阿巴特斯海胆Abatus nimrodi（n=48）的CO1基因区域有2种单倍型，16S基因区域有8种单倍型。此外，数据集还包含戴维斯区域的8只沙克尔顿阿巴特斯海胆A. shackeltoni、4只菲利普阿巴特斯海胆A. philippii及1只洞穴阿巴特斯海胆A. cavernosus样本。

线粒体DNA数据可用信息：数据以4个FASTA文本格式文件存储，分别对应阿巴特斯海胆COI数据、阿巴特斯海胆16S数据、斯特雷奇努斯海胆COI数据。个体编码规则为：前两位字母代表物种（SN=纽迈耶海胆S. neumayeri、AN=尼姆罗德阿巴特斯海胆A. nimrodi、AI=英根斯阿巴特斯海胆A. ingens、AS=沙克尔顿阿巴特斯海胆A. shackletoni、AC=洞穴阿巴特斯海胆A. cavernosus）；接下来两位代表基因区域（CO=COI、16=16S）；戴维斯区域样本后续为3-4位数字，凯西区域样本后续为以41开头的5位数字。

受精实验数据集生成：纽迈耶海胆S. neumayeri样本采集于埃利斯峡湾冰层破裂前。共筛选12个个体用于受精实验（7雄5雌），确保杂交组合的受精成功率可达90%以上。精子在-1.8℃下用过滤海水（FSW）激活，浓度通过血细胞计数板（haemocytometer）测定。实验采用3种温度处理（-1.8℃、1℃、3℃）、3种盐度处理（35ppt、30ppt、25ppt）及5种精卵比（50:1、100:1、500:1、1500:1、2500:1），每个温度-盐度-精卵比组合设置4次重复。30分钟后，每管加入3-5滴10%福尔马林固定卵子以终止受精。受精率测定方式为：每管计数前100个卵子，根据是否存在隆起的受精膜判断其受精状态。

受精实验数据可用信息：数据以Excel文件存储，含3个工作表（对应3种温度处理）。每个工作表包含3个表格（对应3种盐度处理），每个表格含5列，从左至右依次为：精卵比（sperm:egg ratio）、重复编号（rep.No.）、受精卵数（Fert.）、未受精卵数（Unfert.）、平均受精卵数（Mean）。