RegioDiv: Genetic diversity of herbaceous plants in Germany: Agrostis capillaris. RegioDiv Agrostis capillaris

Autochthonous seed material of wild species for ecological restoration is produced in accordance with the rules of the German Regiosaatgut system in 22 seed zones (Bucharova et al. 2019, https://doi.org/10.1007/s10592-018-1067-6). The RegioDiv project analysed genetic diversity of more than 30 grassland plant species (grasses, legumes, non-legume herbs) in all seed zones to test how well seed zones represent genetic patterns. Data on spatial patterns of diversity will be used for practical recommendations for species management and seed zone design. We collected appr. 19,000 leaf samples from 28 taxa in 22 seed zones across Germany, which had been subdivided in 72 subregions. Additional samples were available from neighboring countries. In total 14,496 samples underwent ddRAD analysis following Peterson et al. (2012, https://doi.org/10.1371/journal.pone.0037135) using EcoRI and MspI as restriction enzymes and a selected fragment length of 350-450 bp. Each 1152 samples were sequenced per S4 lane of an Illumina Novaseq 6000 (PE150) with a volume of 2,500 Mio. Sequences, resulting in appr. 2 Mio. Sequences per individual sample. A total of 12,389 samples proved to have a sufficient number of DNA sequences for further analysis. In total 11,762 samples remained after excluding references samples and outgroup species for final data analysis. The number of samples originating in Germany in the final data sets ranged between 89 (Tragopogon orientalis) and 891 (Knautia arvensis), with a mean of 339. This project holds all samples used for Agrostis capillaris

用于生态修复的野生乡土种子材料，严格遵循德国Regiosaatgut（区域种子认证体系）的规则，于22个种子区内完成生产（Bucharova等，2019，https://doi.org/10.1007/s10592-018-1067-6）。RegioDiv项目针对全部22个种子区内的30余种草地植物类群（禾草、豆科植物、非豆科草本植物）的遗传多样性开展分析，旨在验证种子区能否有效反映物种的遗传空间格局。相关多样性空间分布数据，将为物种管理实践与种子区优化设计提供科学参考依据。本研究团队在德国全境22个被细分为72个亚区域的种子区内，共采集了28个分类单元的约19000份叶片样本，此外还获取了部分来自邻国的补充样本。总计14496份样本按照Peterson等（2012，https://doi.org/10.1371/journal.pone.0037135）建立的方法进行ddRAD（双酶切限制性位点相关DNA测序）分析，选用EcoRI与MspI作为限制性内切酶，筛选片段长度介于350-450 bp的测序片段。采用Illumina Novaseq 6000测序平台的S4泳道完成测序，每份泳道可搭载1152份样本进行双端150bp（PE150）测序，单泳道总测序量达25亿条序列，单份样本的平均测序量约为200万条序列。最终共有12389份样本的DNA序列数量与质量满足后续分析的要求。在剔除对照样本与外类群物种后，剩余11762份样本用于最终数据分析。最终数据集中，源自德国的样本数量跨度为89份（东方婆罗门参，Tragopogon orientalis）至891份（田野山萝卜，Knautia arvensis），单物种平均样本量为339份。本项目涵盖了用于细弱剪股颖（Agrostis capillaris）分析的全部样本。