Extending DNA barcoding coverage for Lake Whitefish (Coregonus clupeaformis) across the three major basins of Lake Huron
Lauren M. Overdyk, H. Braid, S. Crawford, R. Hanner
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引用次数: 8
Abstract
DNA barcoding is a useful tool for both species identification and discovery, but the latter requires denser sampling than typically used in barcode studies. Lake Whitefish (Coregonus clupeaformis) is a valuable species, fished traditionally, commercially, and recreationally in Lake Huron. Based on the natural geographic and bathymetric separation of the three major basins in Lake Huron, the potential separation of Lake Whitefish within these basins, and the variation among life history (early and late spawning), we predicted that Lake Huron might harbour cryptic lineages of Lake Whitefish at the basin level. To test this prediction, DNA barcodes of the mitochondrial 5’ cytochrome c oxidase subunit I (COI) gene sequences were recovered from spawning phase Lake Whitefish (n = 5 per site), which were collected from sites (n = 28) around Lake Huron during Fall 2012. These sequences, combined with other publically available DNA barcodes from the Barcode of Life Data System (BOLD), revealed twelve unique haplotypes across North America, with seven unique to Lake Huron. The dominant haplotype was found throughout Lake Huron and east to the St. Lawrence River. No deep divergences were revealed. This comprehensive lake-wide sampling effort offers a new perspective on C. clupeaformis, and Research Article Open Access © 2015 Lauren M. Overdyk et al. licensee De Gruyter Open. This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 License. Lauren M. Overdyk*, Heather E. Braid, Stephen S. Crawford, Robert H. Hanner Extending DNA barcoding coverage for Lake Whitefish (Coregonus clupeaformis) across the three major basins of Lake Huron *Corresponding author:Lauren M. Overdyk, Department of Integrative Biology, University of Guelph, 50 Stone Road East, Guelph, Ontario, Canada N1G 2W1, E-mail: loverdyk@uoguelph.ca Stephen S. Crawford, Robert H. Hanner: Department of Integrative Biology, University of Guelph, 50 Stone Road East, Guelph, Ontario, Canada N1G 2W1 Heather E. Braid: Institute for Applied Ecology New Zealand, Auckland University of Technology, Private Bag 92006, Auckland, New Zealand 1010 Stephen S. Crawford: Chippewas of Nawash, Unceded First Nation, 135 Lakeshore Blvd, Nevaashiinigmiing, Ontario, Canada N1G 2W1 Robert H. Hanner: Biodiversity Institute of Ontario, University of Guelph, 50 Stone Road East, Guelph, Ontario, Canada N1G 2W1 10.1515/dna-2015-0007 Downloaded from De Gruyter Online at 09/12/2016 11:31:59PM via free access 60 L.M. Overdyk et al. 2 Materials and Methods 2.1 Specimen Collection A lake-wide sampling effort for adult Lake Whitefish was undertaken in the fall of 2012 in Lake Huron (Figure 1; Table 1). Spawning-phase Lake Whitefish were sampled from twenty-eight locations across the three basins of Lake Huron: the Main Basin (n = 16), the North Channel (n = 4), and Georgian Bay (n = 8). Early and late samples were taken from Sites 16-18 (Cape Hurd, Stokes Bay, and Howdenvale) along the Western shore of the Saugeen (Bruce) Peninsula based on known variation in spawning activity of Lake Whitefish in these areas (Table 1). Five different individuals, including at least two males and two females, were sampled from each site for DNA barcoding. Two muscle fillets were taken from each individual and maintained at -20oC, thawed briefly for subsampling of approximately 1 gram of tissue for DNA extraction. We had previously obtained representative Lake Whitefish from Stokes Bay and Sharbot Lake, which were processed in the same manner.
在休伦湖的三个主要流域扩展白鱼的DNA条形码覆盖范围
DNA条形码是物种鉴定和发现的有用工具,但后者需要比条形码研究中通常使用的更密集的采样。湖白鱼(Coregonus clupeaformis)是休伦湖传统、商业和娱乐捕捞的珍贵物种。根据休伦湖三个主要盆地的自然地理和水深分离、白鱼在这些盆地内的潜在分离以及生活史(产卵早期和产卵后期)的变化,我们预测休伦湖可能在盆地水平上有白鱼的隐蔽谱系。为了验证这一预测,2012年秋季在休伦湖附近的28个地点收集了白鱼产卵期(n = 5个)的线粒体5 '细胞色素c氧化酶亚基I (COI)基因序列的DNA条形码。这些序列,结合其他来自生命条形码数据系统(BOLD)的公开DNA条形码,揭示了北美12种独特的单倍型,其中7种是休伦湖独有的。优势单倍型在整个休伦湖和圣劳伦斯河以东被发现。没有显示出深刻的分歧。这项全面的全湖采样工作为C. clupeaformis提供了新的视角,研究文章开放获取©2015 Lauren M. Overdyk等人获得De Gruyter Open许可。本作品采用知识共享署名-非商业-禁止衍生3.0许可协议。Lauren M. Overdyk*, Heather E. Braid, Stephen S. Crawford, Robert H. Hanner扩展休伦湖三大流域白鱼(Coregonus clupeaformis)的DNA条形码覆盖*通讯作者:Lauren M. Overdyk,加拿大圭尔夫大学整合生物学系,50 Stone Road East, Guelph, Ontario, Canada N1G 2W1, E-mail: loverdyk@uoguelph.ca加拿大安大略省圭尔夫市石路东50号圭尔夫大学整合生物学系Heather E. Braid:新西兰奥克兰理工大学应用生态研究所,Private Bag 92006,新西兰奥克兰1010 Stephen S. Crawford: Nawash的Chippewas, unded First Nation, 135 lakesshore Blvd, nevaashinigmiing, Ontario, Canada N1G 2W1 Robert H. Hanner:L.M. Overdyk等。2材料与方法2.1标本采集于2012年秋季在休伦湖进行了一项全湖白鱼成虫取样工作(图1;表1)从休伦湖三个流域的28个地点取样产卵期湖白鱼:主要盆地(n = 16)、北海峡(n = 4)和乔治亚湾(n = 8)。根据这些地区已知的湖白鱼产卵活动变化,从索金(布鲁斯)半岛西海岸的16-18个地点(赫德角、斯托克斯湾和豪登维尔)采集了早期和晚期样本(表1)。从每个地点采集了5个不同的个体,包括至少2个雄性和2个雌性,用于DNA条形码。从每个人身上取下两块肌肉片,保存在-20℃,短暂解冻,提取约1克组织进行DNA提取。我们以前从斯托克斯湾和夏博湖获得了代表性的湖白鱼,它们都是以同样的方式加工的。
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