Lara Elena, Holmfeldt Karin, Solonenko Natalie, Sà Elisabet Laia, Ignacio-Espinoza J Cesar, Cornejo-Castillo Francisco M, Verberkmoes Nathan C, Vaqué Dolors, Sullivan Matthew B, Acinas Silvia G
Department of Marine Biology and Oceanography, Institut de Ciències del Mar (CSIC), Passeig Marítim de la Barceloneta, 37-49, 08003 Barcelona, Spain.
University of Arizona, Department of Ecology and Evolutionary Biology, 1007 E. Lowell St., Tucson, AZ, United States of America.
PLoS One. 2015 Jan 14;10(1):e0114829. doi: 10.1371/journal.pone.0114829. eCollection 2015.
Marine viruses (phages) alter bacterial diversity and evolution with impacts on marine biogeochemical cycles, and yet few well-developed model systems limit opportunities for hypothesis testing. Here we isolate phage B8b from the Mediterranean Sea using Pseudoalteromonas sp. QC-44 as a host and characterize it using myriad techniques. Morphologically, phage B8b was classified as a member of the Siphoviridae family. One-step growth analyses showed that this siphovirus had a latent period of 70 min and released 172 new viral particles per cell. Host range analysis against 89 bacterial host strains revealed that phage B8b infected 3 Pseudoalteromonas strains (52 tested, >99.9% 16S rRNA gene nucleotide identity) and 1 non-Pseudoaltermonas strain belonging to Alteromonas sp. (37 strains from 6 genera tested), which helps bound the phylogenetic distance possible in a phage-mediated horizontal gene transfer event. The Pseudoalteromonas phage B8b genome size was 42.7 kb, with clear structural and replication modules where the former were delineated leveraging identification of 16 structural genes by virion structural proteomics, only 4 of which had any similarity to known structural proteins. In nature, this phage was common in coastal marine environments in both photic and aphotic layers (found in 26.5% of available viral metagenomes), but not abundant in any sample (average per sample abundance was 0.65% of the reads). Together these data improve our understanding of siphoviruses in nature, and provide foundational information for a new 'rare virosphere' phage-host model system.
海洋病毒(噬菌体)改变细菌多样性和进化,影响海洋生物地球化学循环,但很少有成熟的模型系统限制了假设检验的机会。在这里,我们以假交替单胞菌属QC-44为宿主,从地中海分离出噬菌体B8b,并使用多种技术对其进行表征。形态学上,噬菌体B8b被归类为长尾噬菌体科的成员。一步生长分析表明,这种长尾噬菌体的潜伏期为70分钟,每个细胞释放172个新的病毒颗粒。对89个细菌宿主菌株的宿主范围分析表明,噬菌体B8b感染了3个假交替单胞菌菌株(测试了52个,16S rRNA基因核苷酸同一性>99.9%)和1个属于交替单胞菌属的非假交替单胞菌菌株(测试了来自6个属的37个菌株),这有助于界定噬菌体介导的水平基因转移事件中可能的系统发育距离。假交替单胞菌噬菌体B8b的基因组大小为42.7 kb,具有明确的结构和复制模块,其中前者通过病毒体结构蛋白质组学鉴定16个结构基因来划定,其中只有4个与已知结构蛋白有任何相似性。在自然界中,这种噬菌体在沿海海洋环境的光合层和无光层都很常见(在26.5%的可用病毒宏基因组中发现),但在任何样本中都不丰富(每个样本的平均丰度为读数的0.