Jantzen Johanna R, Amarasinghe Prabha, Folk Ryan A, Reginato Marcelo, Michelangeli Fabian A, Soltis Douglas E, Cellinese Nico, Soltis Pamela S
Department of Biology University of Florida Gainesville Florida 32611 USA.
Florida Museum of Natural History University of Florida Gainesville Florida 32611 USA.
Appl Plant Sci. 2020 May 9;8(5):e11345. doi: 10.1002/aps3.11345. eCollection 2020 May.
Putatively single-copy nuclear (SCN) loci, which are identified using genomic resources of closely related species, are ideal for phylogenomic inference. However, suitable genomic resources are not available for many clades, including Melastomataceae. We introduce a versatile approach to identify SCN loci for clades with few genomic resources and use it to develop probes for target enrichment in the distantly related and (Melastomataceae).
We present a two-tiered pipeline. First, we identified putatively SCN loci using MarkerMiner and transcriptomes from distantly related species in Melastomataceae. Published loci and genes of functional significance were then added (384 total loci). Second, using HybPiper, we retrieved 689 homologous template sequences for these loci using genome-skimming data from within the focal clades.
We sequenced 193 loci common to and . Probes designed from 56 template sequences successfully targeted sequences in both clades. Probes designed from genome-skimming data within a focal clade were more successful than probes designed from other sources.
Our pipeline successfully identified and targeted SCN loci in and , enabling phylogenomic studies in both clades and potentially across Melastomataceae. This pipeline could be easily applied to other clades with few genomic resources.
利用近缘物种的基因组资源鉴定出的推定单拷贝核(SCN)基因座,是系统发育基因组学推断的理想选择。然而,包括野牡丹科在内的许多分支没有合适的基因组资源。我们引入了一种通用方法,用于为基因组资源较少的分支鉴定SCN基因座,并利用该方法开发用于远缘的 和 (野牡丹科)目标富集的探针。
我们提出了一个两层管道。首先,我们使用MarkerMiner和野牡丹科远缘物种的转录组鉴定推定的SCN基因座。然后添加已发表的基因座和具有功能意义的基因(共384个基因座)。其次,使用HybPiper,我们利用来自目标分支内的基因组浅层测序数据检索了这些基因座的689个同源模板序列。
我们对 和 共有的193个基因座进行了测序。从56个模板序列设计的探针成功靶向了两个分支中的序列。从目标分支内的基因组浅层测序数据设计的探针比从其他来源设计的探针更成功。
我们的管道成功鉴定并靶向了 和 的SCN基因座,使两个分支以及可能整个野牡丹科的系统发育基因组学研究成为可能。该管道可以很容易地应用于其他基因组资源较少的分支。