Marine Genomics Unit, Okinawa Institute of Science and Technology Graduate University, 1919-1 Tancha, Onna, Okinawa 904-0495, Japan.
Natural Science Center for Basic Research and Development, Gene Science Division, Hiroshima University, 1-4-2 Kagamiyama, Higashi-Hiroshima, Hiroshima 739-8527, Japan.
Gigascience. 2019 Apr 1;8(4). doi: 10.1093/gigascience/giz023.
Acoels are primitive bilaterians with very simple soft bodies, in which many organs, including the gut, are not developed. They provide platforms for studying molecular and developmental mechanisms involved in the formation of the basic bilaterian body plan, whole-body regeneration, and symbiosis with photosynthetic microalgae. Because genomic information is essential for future research on acoel biology, we sequenced and assembled the nuclear genome of an acoel, Praesagittifera naikaiensis.
To avoid sequence contamination derived from symbiotic microalgae, DNA was extracted from embryos that were free of algae. More than 290x sequencing coverage was achieved using a combination of Illumina (paired-end and mate-pair libraries) and PacBio sequencing. RNA sequencing and Iso-Seq data from embryos, larvae, and adults were also obtained. First, a preliminary ∼17-kilobase pair (kb) mitochondrial genome was assembled, which was deleted from the nuclear sequence assembly. As a result, a draft nuclear genome assembly was ∼656 Mb in length, with a scaffold N50 of 117 kb and a contig N50 of 57 kb. Although ∼70% of the assembled sequences were likely composed of repetitive sequences that include DNA transposons and retrotransposons, the draft genome was estimated to contain 22,143 protein-coding genes, ∼99% of which were substantiated by corresponding transcripts. We could not find horizontally transferred microalgal genes in the acoel genome. Benchmarking Universal Single-Copy Orthologs analyses indicated that 77% of the conserved single-copy genes were complete. Pfam domain analyses provided a basic set of gene families for transcription factors and signaling molecules.
Our present sequencing and assembly of the P. naikaiensis nuclear genome are comparable to those of other metazoan genomes, providing basic information for future studies of genic and genomic attributes of this animal group. Such studies may shed light on the origins and evolution of simple bilaterians.
后生动物是具有非常简单柔软身体的原始两侧对称动物,其中许多器官,包括肠道,都没有发育。它们为研究涉及基本两侧对称动物体节形成、全身再生和与光合微藻共生的分子和发育机制提供了平台。由于基因组信息对于后生动物生物学的未来研究至关重要,我们对后生动物 Praesagittifera naikaiensis 的核基因组进行了测序和组装。
为了避免来自共生微藻的序列污染,从没有藻类的胚胎中提取 DNA。使用 Illumina(配对末端和 mate-pair 文库)和 PacBio 测序的组合,实现了超过 290x 的测序覆盖率。还获得了来自胚胎、幼虫和成虫的 RNA 测序和 Iso-Seq 数据。首先,组装了一个初步的约 17 千碱基对(kb)线粒体基因组,该基因组从核序列组装中删除。结果,一个草稿核基因组组装长约 656Mb,支架 N50 为 117kb,串联 N50 为 57kb。尽管约 70%的组装序列可能由包括 DNA 转座子和逆转座子在内的重复序列组成,但该草案基因组估计包含 22143 个蛋白质编码基因,其中约 99%有相应的转录本支持。我们在后生动物基因组中未发现水平转移的微藻基因。基准通用单拷贝同源物分析表明,77%的保守单拷贝基因是完整的。Pfam 结构域分析为转录因子和信号分子提供了基本的基因家族。
我们目前对 P. naikaiensis 核基因组的测序和组装与其他后生动物基因组相当,为该动物类群的基因和基因组属性的未来研究提供了基本信息。这些研究可能有助于揭示简单两侧对称动物的起源和进化。