Friedrich Ines, Neubauer Hannes, Kuritsyn Alisa, Bodenberger Bernhard, Tskhay Faina, Hartmann Sara, Poehlein Anja, Bömeke Mechthild, Hoppert Michael, Schneider Dominik, Hertel Robert, Daniel Rolf
Genomic and Applied Microbiology, Göttingen Genomics Laboratory, Institute of Microbiology and Genetics, Georg-August University of Göttingen, Göttingen, Germany.
General Microbiology, Institute of Microbiology and Genetics, Georg-August University of Göttingen, Göttingen, Germany.
Front Microbiol. 2023 Mar 21;14:1095850. doi: 10.3389/fmicb.2023.1095850. eCollection 2023.
Focusing on visible plaques for phage isolation leaves the question if we miss the diversity of non-plaque forming phages. We addressed this question through direct plaque-based isolation by employing the new hosts LVF1 and LVF3 dsDNA, ssDNA, dsRNA, and ssRNA host-associated metavirome analysis. Of the 25 distinctive dsDNA phage isolates, 14 were associated with and 11 with . TEM analysis revealed that 6 were myoviruses, 18 siphoviruses and 1 podovirus, while phages infecting belonged all to siphoviruses. The associated viromes suggested a higher phage diversity in summer than in winter, and dsDNA phages were the dominant group. Isolation of vB_SmaP-Kaonashi was possible after investigating the viromes associated with , demonstrating the great potential of accompanying host-associated metavirome analysis. The ssDNA virome analysis showed that the LVF1 host is associated with and phages, although none of them were isolated. The results demonstrated that the classical isolation technique is not exhausted, leading to the isolation of new dsDNA phages. It can be further improved by combination with metavirome techniques, which revealed further diversity.
专注于可见噬菌斑进行噬菌体分离,会引发一个问题,即我们是否遗漏了非噬菌斑形成噬菌体的多样性。我们通过使用新宿主LVF1和LVF3进行基于噬菌斑的直接分离,对dsDNA、ssDNA、dsRNA和ssRNA宿主相关宏病毒组进行分析,从而解决了这个问题。在25种独特的dsDNA噬菌体分离株中,14种与……相关,11种与……相关。透射电子显微镜分析显示,6种是肌尾噬菌体,18种是长尾噬菌体,1种是短尾噬菌体,而感染……的噬菌体均属于长尾噬菌体。相关的病毒组表明,夏季的噬菌体多样性高于冬季,且dsDNA噬菌体是主要群体。在研究与……相关的病毒组后,成功分离出了vB_SmaP-Kaonashi,这证明了伴随宿主相关宏病毒组分析具有巨大潜力。单链DNA病毒组分析表明,LVF1宿主与……和……噬菌体相关,尽管它们均未被分离出来。结果表明,经典的分离技术尚未穷尽,仍能分离出新的dsDNA噬菌体。通过与宏病毒组技术相结合,可进一步改进该技术,从而揭示更多的多样性。