In silico and empirical quantification of viruses is paramount for obtaining information on viral populations that have a major impact on biogeochemical cycles. The uncultured Pelagibacter virus vSAG 37‐F6 discovered via single‐virus genomics is one of the most abundant and cosmopolitan marine viruses; however, little is understood about its temporal variation. Here, we estimated the absolute number of infecting 37‐F6 viruses in coastal bacterioplankton from the Mediterranean Sea by using a novel, feasible SYBR Green I chip‐based digital PCR (SYBR dPCR) technique, not implemented before for enumerating (uncultured) microbes. Quantitative SYBR dPCR estimated 450–3480 genome copies of virus 37‐F6 in cells/mL (i.e. infecting viruses) and a total of ≈10–400 putative infected cells/mL with a potential C release of 0.12–4.9 pg/ml in the analysed samples. Considering that virus 37‐F6 is ubiquitous and abundant in all Tara samples, an enormous amount of C could be transformed by this virus through the ‘viral shunt’. Thus, this SYBR dPCR technique has enabled the absolute quantification of an ecologically relevant uncultured virus in nature and the estimation of its potential contribution on biogeochemical cycles. Overall, our study also shows that this approach has a broad applicability for quantifying any other target loci in Microbiology and Virology.

中文翻译：

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通过基于芯片的数字聚合酶链反应绝对定量感染病毒颗粒。

在计算机上对于获得对生物地球化学循环有重大影响的病毒种群信息，病毒的经验量化至关重要。通过单病毒基因组学发现的未经培养的杆菌病毒vSAG 37-F6是最丰富和国际化的海洋病毒之一；然而，对其时间变化知之甚少。在这里，我们通过使用一种新颖的，可行的基于SYBR Green I芯片的数字PCR（SYBR dPCR）技术估算了地中海沿岸浮游细菌感染37-F6病毒的绝对数量，该技术以前并未用于枚举（未培养的）微生物。定量SYBR dPCR估计在细胞/ mL（即感染病毒）中病毒450-3480基因组拷贝数为37-F6（即感染病毒），在总共约10-400个推定感染细胞/ mL中，潜在的C释放量为0.12-4.9 pg / ml。分析样品。在Tar​​a样品中，这种病毒可以通过“病毒分流器”转化大量C。因此，这种SYBR dPCR技术已经能够对自然界中与生态相关的未培养病毒进行绝对定量，并能够估计其对生物地球化学循环的潜在贡献。总体而言，我们的研究还表明，这种方法对于定量微生物学和病毒学中的任何其他目标基因座具有广泛的适用性。