Poretsky Rachel S, Hewson Ian, Sun Shulei, Allen Andrew E, Zehr Jonathan P, Moran Mary Ann
University of Georgia, Department of Marine Sciences, Athens, GA 30602, USA.
Environ Microbiol. 2009 Jun;11(6):1358-75. doi: 10.1111/j.1462-2920.2008.01863.x. Epub 2009 Feb 4.
Metatranscriptomic analyses of microbial assemblages (< 5 microm) from surface water at the Hawaiian Ocean Time-Series (HOT) revealed community-wide metabolic activities and day/night patterns of differential gene expression. Pyrosequencing produced 75 558 putative mRNA reads from a day transcriptome and 75 946 from a night transcriptome. Taxonomic binning of annotated mRNAs indicated that Cyanobacteria contributed a greater percentage of the transcripts (54% of annotated sequences) than expected based on abundance (35% of cell counts and 21% 16S rRNA of libraries), and may represent the most actively transcribing cells in this surface ocean community in both the day and night. Major heterotrophic taxa contributing to the community transcriptome included alpha-Proteobacteria (19% of annotated sequences, most of which were SAR11-related) and gamma-Proteobacteria (4%). The composition of transcript pools was consistent with models of prokaryotic gene expression, including operon-based transcription patterns and an abundance of genes predicted to be highly expressed. Metabolic activities that are shared by many microbial taxa (e.g. glycolysis, citric acid cycle, amino acid biosynthesis and transcription and translation machinery) were well represented among the community transcripts. There was an overabundance of transcripts for photosynthesis, C1 metabolism and oxidative phosphorylation in the day compared with night, and evidence that energy acquisition is coordinated with solar radiation levels for both autotrophic and heterotrophic microbes. In contrast, housekeeping activities such as amino acid biosynthesis, membrane synthesis and repair, and vitamin biosynthesis were overrepresented in the night transcriptome. Direct sequencing of these environmental transcripts has provided detailed information on metabolic and biogeochemical responses of a microbial community to solar forcing.
对来自夏威夷海洋时间序列(HOT)表层水的微生物群落(<5微米)进行的宏转录组分析揭示了群落范围内的代谢活动以及基因表达差异的昼夜模式。焦磷酸测序从白天转录组产生了75558条假定的mRNA读数,从夜间转录组产生了75946条。对注释mRNA的分类分箱表明,蓝细菌在转录本中所占的百分比(注释序列的54%)高于基于丰度预期的比例(细胞计数的35%和文库16S rRNA的21%),并且可能代表了这个表层海洋群落中白天和夜间转录最活跃的细胞。对群落转录组有贡献的主要异养类群包括α-变形菌纲(注释序列的19%,其中大部分与SAR11相关)和γ-变形菌纲(4%)。转录本库的组成与原核基因表达模型一致,包括基于操纵子的转录模式以及大量预计高表达的基因。许多微生物类群共有的代谢活动(如糖酵解、柠檬酸循环、氨基酸生物合成以及转录和翻译机制)在群落转录本中得到了很好的体现。与夜间相比,白天光合作用、C1代谢和氧化磷酸化的转录本过多,并且有证据表明自养和异养微生物的能量获取与太阳辐射水平相协调。相比之下,氨基酸生物合成、膜合成与修复以及维生素生物合成等管家活动在夜间转录组中占比过高。对这些环境转录本的直接测序提供了有关微生物群落对太阳强迫的代谢和生物地球化学响应的详细信息。