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Characterization and genomic analysis of a plaque purified strain of cyanophage PP

Virologica Sinica volume 28, pages 272–279 (2013)Cite this article

Abstract

Cyanophages are ubiquitous and essential components of the aquatic environment and play an important role in the termination of algal blooms. As such, they have attracted widespread interest. PP was the first isolated cyanophage in China, which infects Plectonema boryanum and Phormidium foveolarum. In this study, this cyanophage was purified three times by a double-agar overlay plaque assay and characterized. Its genome was extracted, totally sequenced and analyzed. Electron microscopy revealed a particle with an icosahedral head connected to a short stubby tail. Bioassays showed that PP was quite virulent. The genome of PP is a 42,480 base pair (bp), linear, double-stranded DNA molecule with 222 bp terminal repeats. It has high similarity with the known Pf-WMP3 sequence. It contains 41 open reading frames (ORFs), 17 of which were annotated. Intriguingly, the genome can be divided into two completely different parts, which differ both in orientation and function.

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References

  • Bergh O, Borsheim K Y, Bratbak G, et al. 1989. High abundance of viruses found in aquatic environments. Nature, 340: 467–468.

    PubMed  Article  CAS  Google Scholar 

  • Bratbak G, Heldal M, Norland S, et al. 1990. Viruses as partners in spring bloom microbial trophodynamics. Appl Environ Microbiol, 56(5): 1400–1405.

    PubMed  CAS  Google Scholar 

  • Bryan M J, Burroughs N J, Spence E M, et al. 2008. Evidence for the intense exchange of MazG in marine cyanophages by horizontal gene transfer. PloS One, 3(4): e2048.

    PubMed  Article  Google Scholar 

  • Carmichael W W. 2001. Health effects of toxin-producing Cyanobacteria: “The CyanoHABs”. Hum Ecol Risk Assess, 7(5): 1393–1407.

    Article  Google Scholar 

  • Chenard C, and Suttle C A. 2008. Phylogenetic diversity of sequences of cyanophage photosynthetic gene psbA in marine and freshwaters. Appl Environ Microbiol, 74(17): 5317–5324.

    PubMed  Article  CAS  Google Scholar 

  • Chen F, and Lu J R. 2002. Genomic sequence and evolution of marine Cyanophage P60: a new insight on lytic and lysogenic phages. Appl Environ Microbiol, 68(5): 2589–2594.

    PubMed  Article  CAS  Google Scholar 

  • Codd G A, Morrison L F, and Metcalf J S. 2005. Cyanobacterial toxins: risk management for health protection. Toxicol Appl Pharmacol, 203: 264–272.

    PubMed  Article  CAS  Google Scholar 

  • Dittmann E, and Wiegand C. 2006. Cyanobacterial toxins-occurrence, biosynthesis and impact on human affairs. Mol Nutr Food Res, 50: 7–17.

    PubMed  Article  CAS  Google Scholar 

  • Fuhrman J A. 1999. Marine viruses and their biogeochemical and ecological effects. Nature, 399: 541–548.

    PubMed  Article  CAS  Google Scholar 

  • Gao E B, Gui J F, and Zhang Q Y. 2012. A novel cyanophage with cyanobacterial non-bleaching protein A gene in the genome. J Virol, 86(1): 236–245.

    PubMed  Article  CAS  Google Scholar 

  • Jungblut A D, Hawes I, Mountfort D, et al. 2005. Diversity within cyanobacterial mat communities in variable salinity meltwater ponds of McMurdo Ice Shelf, Antarctica. Environ Microbiol, 7(4): 519–529.

    PubMed  Article  CAS  Google Scholar 

  • Kropinski A M, Mazzocco A, Waddell TE, et al. 2009. Enumeration of bacteriophages by double agar overlay plaque assay. Methods Mol Biol, 501: 69–76.

    PubMed  Article  CAS  Google Scholar 

  • Liao M J, Cheng K, Yang J Y, et al. 2010. Assessment of UV-B damage in cyanophage PP. Aquat Microb Ecol, 58: 323–328.

    Article  Google Scholar 

  • Liu X Y, Shi M, Kong S L, et al. 2007. Cyanophage Pf-WMP4, a T7-like phage infecting the freshwater cyanobacterium Phormidium foveolarum: Complete genome sequence and DNA translocation. Virology, 366: 28–39.

    PubMed  Article  CAS  Google Scholar 

  • Liu X Y, Kong S L, Shi M, et al. 2008. Genomic analysis of freshwater cyanophage Pf-WMP3 infecting cyanobacterium Phormidium foveolarum: the conserved elements for a phage. Microb Ecol, 56: 671–680.

    PubMed  Article  CAS  Google Scholar 

  • Lorraine C B. 2002. Cyanobacterial Harmful Algal Blooms (CyanoHABs): Developing a public health response. Lake Reserv Manag, 18(1): 20–31.

    Article  Google Scholar 

  • Mann N H, Clokie M R J, Millard A, et al. 2005. The genome of S-PM2, a “photosynthetic” T4-type bacteriophage that infects marine Synechococcus strains. J Bacteriol, 187(9): 3188–3200.

    PubMed  Article  CAS  Google Scholar 

  • Paul J H, and Weinbauer M. 2010. Detection of lysogeny in marine environments. In Wilhelm S W, Weinbauer M G, and Suttle C A (eds), Manual of Aquatic Viral Ecology, pp 30–33, American Society of Limnology and Oceanography, TX, USA.

    Chapter  Google Scholar 

  • Pope W H, Weigele P R, Chang J, et al. 2007. Genome sequence, structural proteins, and capsid organization of the cyanophage Syn5: a “horned” bacteriophage of marine Synechococcus. J Mol Biol, 368: 966–981.

    PubMed  Article  CAS  Google Scholar 

  • Proctor L M, and Fuhrman J A. 1990. Viral mortality of marine bacteria and cyanobacteria. Nature, 343: 60–62.

    Article  Google Scholar 

  • Raytcheva D A, Haase-Pettingell C, Piret J, et al. 2011. Intracellular assembly of cyanophage Syn5 proceeds through a scaffold-containing procapsid. J Virol, 85(5): 2406–2415.

    PubMed  Article  CAS  Google Scholar 

  • Sabehi G, Shaulov L, Silver D H, et al. 2012. A novel lineage of myoviruses infecting cyanobacteria is widespread in the oceans. Proc Natl Acad Sci U S A, 109(6): 2037–2042.

    PubMed  Article  CAS  Google Scholar 

  • Safferman R S, and Morris S E. 1963. Algal virus: isolation. Science, 140(3567): 679–680.

    PubMed  Article  CAS  Google Scholar 

  • Safferman R S. 1983. Classification and nomenclature of viruses of cyanobacteria. Intervirology, 19: 61–66.

    PubMed  Article  CAS  Google Scholar 

  • Sherman L A, Haselkorn R. 1970. LPP-1 infection of the blue-green alga Plectonema boryanum: I. Electron Microscopy. J Virol, 6(6): 820–833.

    PubMed  CAS  Google Scholar 

  • Sullivan M B, Coleman M L, Weigele P, et al. 2005. Three Prochlorococcus cyanophage genomes: signature features and ecological interpretations. PloS Biol, 3(5): e144.

    PubMed  Article  Google Scholar 

  • Sullivan M B, Krastins B, Hughes J L, et al. 2009. The genome and structural proteome of an ocean siphovirus: A new window into the cyanobacterial ‘mobilome’. Environ Microbiol, 11: 2935–2951.

    PubMed  Article  CAS  Google Scholar 

  • Suttle CA. 2005. Viruses in the sea. Nature, 437: 356–361.

    PubMed  Article  CAS  Google Scholar 

  • Suttle CA. 2007. Marine viruses-major players in the global ecosystem. Nat Rev Microbiol, 5: 801–812.

    PubMed  Article  CAS  Google Scholar 

  • Weigele P R, Pope W H, Pedulla M L, et al. 2007. Genomic and structural analysis of Syn9, a cyanophage infecting marine Prochlorococcus and Synechococcus. Environ Microbiol, 9: 1675–1695.

    PubMed  Article  CAS  Google Scholar 

  • Wilhelm S W, Carberry M J, Eldridge M L, et al. 2006. Marine and freshwater cyanophages in a Laurentian Great Lake: evidence from infectivity assays and molecular analyses of g20 genes. Appl Environ Microbiol, 72(7): 4957–4963.

    PubMed  Article  CAS  Google Scholar 

  • Wilson W H, Joint L R, Carr N G, et al. 1993. Isolation and molecular characterization of five marine cyanophages propagated on S ynechococcus sp. strain WH7803. Appl Environ Microbiol, 59: 3736–3743.

    PubMed  CAS  Google Scholar 

  • Yoshida M, Yoshida T, Kashima A, et al. 2008. Ecological dynamics of the toxic bloom-forming cyanobacterium Microcystis aeruginosa and its cyanophages in freshwater. Appl Environ Microbiol, 74(10): 3269–3273.

    PubMed  Article  CAS  Google Scholar 

  • Yoshida T, Nagasaki K, Takashima Y, et al. 2008. Ma-LMM01 infecting Microcystis aeruginosa illuminates diverse cyanophage genome strategies. J Bacteriol, 190(5): 1762–1772.

    PubMed  Article  CAS  Google Scholar 

  • Zhao Y J, Chen K, Shi Z L, et al. 2002. Isolation and identification of the first cyanophage in China. Prog Nat Sci, 12: 923–927. (In Chinese)

    Google Scholar 

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Authors and Affiliations

  1. State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China

    Yiran Zhou, Juan Lin, Na Li, Zhihong Hu & Fei Deng

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  1. Yiran Zhou
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  2. Juan Lin
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  3. Na Li
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  4. Zhihong Hu
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  5. Fei Deng
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Correspondence to Fei Deng.

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Zhou, Y., Lin, J., Li, N. et al. Characterization and genomic analysis of a plaque purified strain of cyanophage PP. Virol. Sin. 28, 272–279 (2013). https://doi.org/10.1007/s12250-013-3363-0

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  • DOI: https://doi.org/10.1007/s12250-013-3363-0

Keywords

  • Cyanophage PP
  • Characterization
  • Plaque assay
  • Complete genome sequencing
  • Genome organization