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New possibilities arise for studies of hybridization: SNP-based markers for the multi-species Daphnia longispina complex derived from transcriptome data

机译:杂交研究的新可能性出现:基于转录组数据的多物种水蚤(Daphnia longispina)复合物的基于SNP的标记

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摘要

In order to trace community dynamics and reticulate evolution in hybrid species complexes, long-term comparative studies of natural populations are necessary. Such studies require the development of tools for fine-scale genetic analyses. In the present study, we developed species-diagnostic SNP-based markers for hybridizing freshwater crustaceans: the multispecies Daphnia longispina complex. Specifically, we took advantage of transcriptome data from a key species of this hybrid complex, the annotated genome of a related Daphnia species and well-defined reference genotypes from three parental species. Altogether eleven nuclear loci with several species-specific SNP sites were identified in sequence alignments of these reference genotypes from three parental species and their interspecific hybrids. A PCR-RFLP assay was developed for cost-efficient large population screening by SNP-based genotyping. Taxon assignment by RFLP patterns was nearly perfectly concordant with microsatellite genotyping across several screened populations from Europe. Finally, we were able to amplify two short regions of these loci in formaldehyde-preserved samples dating back to the year 1960. The species-specific SNP-based markers developed here provide valuable tools to study hybridization over time, including the long-term impact of various environmental factors on hybridization and biodiversity changes. SNP-based genotyping will finally allow eco-evolutionary dynamics to be revealed at different time scales.
机译:为了追踪杂种物种复合体中的群落动态和网状进化,必须对自然种群进行长期比较研究。此类研究需要开发用于精细遗传分析的工具。在本研究中,我们开发了基于物种可诊断SNP的杂交淡水甲壳类动物的标记物:多物种水蚤(Daphnia longispina)复合物。具体来说,我们利用了该杂种复合体关键物种的转录组数据,相关水蚤物种的注释基因组以及来自三个亲本物种的明确定义的参考基因型。在来自三个亲本物种及其种间杂种的这些参考基因型的序列比对中,总共鉴定出具有几个物种特异性SNP位点的11个核基因座。通过基于SNP的基因分型,开发了一种经济高效的大批人群筛选PCR-RFLP分析方法。通过RFLP模式进行的分类群分配与来自欧洲多个筛选人群的微卫星基因分型几乎完全一致。最后,我们能够在1960年以前保存甲醛的样本中扩增这些基因座的两个短区域。此处开发的基于物种的基于SNP的特定标记物为研究随时间推移的杂交提供了有价值的工具,包括长期影响环境因素对杂交和生物多样性变化的影响基于SNP的基因分型将最终允许在不同的时间尺度上揭示生态进化动力学。

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