Song Yizhi, Kaster Anne-Kristin, Vollmers John, Song Yanqing, Davison Paul A, Frentrup Martinique, Preston Gail M, Thompson Ian P, Murrell J Colin, Yin Huabing, Hunter C Neil, Huang Wei E
Department of Engineering Science, University of Oxford, Parks Road, Oxford, OX1 3PJ, UK.
Leibniz Institute DSMZ, Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Inhoffenstrasse 7 B, 38124, Braunschweig, Germany.
Microb Biotechnol. 2017 Jan;10(1):125-137. doi: 10.1111/1751-7915.12420. Epub 2016 Oct 17.
Cell sorting coupled with single-cell genomics is a powerful tool to circumvent cultivation of microorganisms and reveal microbial 'dark matter'. Single-cell Raman spectra (SCRSs) are label-free biochemical 'fingerprints' of individual cells, which can link the sorted cells to their phenotypic information and ecological functions. We employed a novel Raman-activated cell ejection (RACE) approach to sort single bacterial cells from a water sample in the Red Sea based on SCRS. Carotenoids are highly diverse pigments and play an important role in phototrophic bacteria, giving strong and distinctive Raman spectra. Here, we showed that individual carotenoid-containing cells from a Red Sea sample were isolated based on the characteristic SCRS. RACE-based single-cell genomics revealed putative novel functional genes related to carotenoid and isoprenoid biosynthesis, as well as previously unknown phototrophic microorganisms including an unculturable Cyanobacteria spp. The potential of Raman sorting coupled to single-cell genomics has been demonstrated.
细胞分选与单细胞基因组学相结合是一种强大的工具，可规避微生物培养并揭示微生物“暗物质”。单细胞拉曼光谱（SCRS）是单个细胞的无标记生化“指纹”，可将分选的细胞与其表型信息和生态功能联系起来。我们采用了一种新颖的拉曼激活细胞喷射（RACE）方法，基于SCRS从红海的水样中分选单个细菌细胞。类胡萝卜素是高度多样的色素，在光合细菌中起重要作用，产生强烈且独特的拉曼光谱。在这里，我们表明基于特征性SCRS从红海样本中分离出了单个含类胡萝卜素的细胞。基于RACE的单细胞基因组学揭示了与类胡萝卜素和类异戊二烯生物合成相关的推定新功能基因，以及以前未知的光合微生物，包括一种不可培养的蓝藻属。已证明拉曼分选与单细胞基因组学相结合的潜力。
