National Science Foundation, 2415 Eisenhower Ave., Alexandria, VA, 22314, USA.
Bigelow Laboratory for Ocean Sciences, 60 Bigelow Drive, East Boothbay, ME, 04544, USA.
Sci Rep. 2019 Apr 15;9(1):6025. doi: 10.1038/s41598-019-42487-1.
Marine planktonic protists are critical components of ocean ecosystems and are highly diverse. Molecular sequencing methods are being used to describe this diversity and reveal new associations and metabolisms that are important to how these ecosystems function. We describe here the use of the single cell genomics approach to sample and interrogate the diversity of the smaller (pico- and nano-sized) protists from a range of oceanic samples. We created over 900 single amplified genomes (SAGs) from 8 Tara Ocean samples across the Indian Ocean and the Mediterranean Sea. We show that flow cytometric sorting of single cells effectively distinguishes plastidic and aplastidic cell types that agree with our understanding of protist phylogeny. Yields of genomic DNA with PCR-identifiable 18S rRNA gene sequence from single cells was low (15% of aplastidic cell sorts, and 7% of plastidic sorts) and tests with alternate primers and comparisons to metabarcoding did not reveal phylogenetic bias in the major protist groups. There was little evidence of significant bias against or in favor of any phylogenetic group expected or known to be present. The four open ocean stations in the Indian Ocean had similar communities, despite ranging from 14°N to 20°S latitude, and they differed from the Mediterranean station. Single cell genomics of protists suggests that the taxonomic diversity of the dominant taxa found in only several hundreds of microliters of surface seawater is similar to that found in molecular surveys where liters of sample are filtered.
海洋浮游原生生物是海洋生态系统的关键组成部分,具有高度多样性。分子测序方法被用于描述这种多样性,并揭示新的关联和新陈代谢,这些对这些生态系统的功能非常重要。我们在这里描述了使用单细胞基因组学方法来采样和研究来自一系列海洋样本的较小(微微型和纳米级)原生生物的多样性。我们从印度洋和地中海的 8 个 Tara 海洋样本中创建了超过 900 个单细胞扩增基因组(SAG)。我们表明,单细胞流式细胞术分选有效地区分了具有叶绿体和无叶绿体的细胞类型,这与我们对原生生物系统发育的理解一致。从单个细胞中获得具有可PCR 鉴定的 18S rRNA 基因序列的基因组 DNA 的产量很低(无叶绿体细胞分选的 15%,叶绿体细胞分选的 7%),并且使用替代引物的测试和与代谢组学的比较并未显示出主要原生生物群中存在系统发育偏见。几乎没有证据表明存在对任何预期或已知存在的系统发育群有利或不利的明显偏见。印度洋的四个公海站尽管从北纬 14°到南纬 20°,但其群落相似,与地中海站不同。原生生物的单细胞基因组学表明,在仅几百微升的表层海水中发现的优势分类群的分类多样性与在过滤数升样本的分子调查中发现的多样性相似。