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Abstract

Bacilladnaviruses are single-stranded DNA viruses that infect diatoms that, so far, have been primarily identified in marine organisms and environments. Using a viral metagenomics approach, we discovered 13 novel bacilladnaviruses originating from samples of mud-flat snail (; =3 genomes) and benthic sediments (=10 genomes) collected from the Avon-Heathcote Estuary in New Zealand. Comparative genomics and phylogenetic analysis of the new bacilladnavirus sequences in the context of the previously classified members of the family helped refine and further expand the taxonomy. Here, based on the replication-associated protein phylogeny and pairwise identities, we established 4 new genera – , , and – and 13 new species within the family. Comparison of the bacilladnavirus capsid protein sequences suggests that the positively charged N-terminal region (R-arm) is required for encapsidation of the larger genomes, whereas the smaller bacilladnavirus genomes can be packaged in the absence of the R-arm subdomain. Furthermore, analysis of the bacilladnavirus genomes revealed that members of three genera encode a highly derived variant of a phospholipase A1, which is predicted to be involved in the lysis of the infected diatoms and/or facilitates the entry of the virions into the host cells. Collectively, our results allow refining of the taxonomy of bacilladnaviruses and provide new insights into the biology and evolution of this understudied group of diatom viruses.

Funding
This study was supported by the:
  • The Brian Mason Scientific & Technical Trust funds
    • Principal Award Recipient: ArvindVarsani
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. The Microbiology Society waived the open access fees for this article.
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2025-03-12
2026-01-14

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References

  1. Krupovic M, Varsani A, Kazlauskas D, Breitbart M, Delwart E et al. Cressdnaviricota: a virus phylum unifying seven families of rep-encoding viruses with single-stranded, circular DNA genomes. J Virol 2020; 94:e00582-20 [View Article] [PubMed]
    [Google Scholar]
  2. Kazlauskas D, Dayaram A, Kraberger S, Goldstien S, Varsani A et al. Evolutionary history of ssDNA bacilladnaviruses features horizontal acquisition of the capsid gene from ssRNA nodaviruses. Virology 2017; 504:114–121 [View Article] [PubMed]
    [Google Scholar]
  3. Kazlauskas D, Varsani A, Krupovic M. Pervasive chimerism in the replication-associated proteins of uncultured single-stranded DNA viruses. Viruses 2018; 10:187 [View Article] [PubMed]
    [Google Scholar]
  4. Munke A, Kimura K, Tomaru Y, Wang H, Yoshida K et al. Primordial capsid and spooled ssDNA genome structures unravel ancestral events of eukaryotic viruses. mBio 2022; 13:e0015622 [View Article] [PubMed]
    [Google Scholar]
  5. Nagasaki K, Tomaru Y, Takao Y, Nishida K, Shirai Y et al. Previously unknown virus infects marine diatom. Appl Environ Microbiol 2005; 71:3528–3535 [View Article] [PubMed]
    [Google Scholar]
  6. Tomaru Y, Shirai Y, Suzuki H, Nagasaki T, Nagumo T. Isolation and characterization of a new single-stranded DNA virus infecting the cosmopolitan marine diatom Chaetoceros debilis. Aquat Microb Ecol 2008; 50:103–112 [View Article]
    [Google Scholar]
  7. Tomaru Y, Shirai Y, Toyoda K, Nagasaki K. Isolation and characterisation of a single-stranded DNA virus infecting the marine planktonic diatom Chaetoceros tenuissimus. . Aquat Microb Ecol 2011; 64:175–184 [View Article]
    [Google Scholar]
  8. Kimura K, Tomaru Y. Isolation and characterization of a single-stranded DNA virus infecting the marine diatom Chaetoceros sp. strain SS628-11 isolated from western Japan. PLoS One 2013; 8:e82013 [View Article] [PubMed]
    [Google Scholar]
  9. Tomaru Y, Takao Y, Suzuki H, Nagumo T, Koike K et al. Isolation and characterization of a single-stranded DNA virus infecting Chaetoceros lorenzianus grunow. Appl Environ Microbiol 2011; 77:5285–5293 [View Article] [PubMed]
    [Google Scholar]
  10. Gastineau R, Lemieux C, Turmel M, Grypioti E, Verret F et al. Two new bacilladnaviruses associated with the diatom Haslea ostrearia. Eur J Phycol 2020; 55:444–453 [View Article]
    [Google Scholar]
  11. Tomaru Y, Toyoda K, Suzuki H, Nagumo T, Kimura K et al. New single-stranded DNA virus with a unique genomic structure that infects marine diatom Chaetoceros setoensis. Sci Rep 2013; 3:3337 [View Article] [PubMed]
    [Google Scholar]
  12. Kimura K, Tomaru Y. Discovery of two novel viruses expands the diversity of single-stranded DNA and single-stranded RNA viruses infecting a cosmopolitan marine diatom. Appl Environ Microbiol 2015; 81:1120–1131 [View Article] [PubMed]
    [Google Scholar]
  13. Tisza MJ, Pastrana DV, Welch NL, Stewart B, Peretti A et al. Discovery of several thousand highly diverse circular DNA viruses. Elife 2020; 9:e51971 [View Article] [PubMed]
    [Google Scholar]
  14. McDaniel LD, Rosario K, Breitbart M, Paul JH. Comparative metagenomics: natural populations of induced prophages demonstrate highly unique, lower diversity viral sequences. Environ Microbiol 2014; 16:570–585 [View Article] [PubMed]
    [Google Scholar]
  15. King AMQ, Lefkowitz EJ, Mushegian AR, Adams MJ, Dutilh BE et al. Changes to taxonomy and the international code of virus classification and nomenclature ratified by the international committee on taxonomy of viruses (2018). Arch Virol 2018; 163:2601–2631 [View Article] [PubMed]
    [Google Scholar]
  16. Zerbini FM, Siddell SG, Lefkowitz EJ, Mushegian AR, Adriaenssens EM et al. Changes to virus taxonomy and the ICTV statutes ratified by the international committee on taxonomy of viruses (2023). Arch Virol 2023; 168:175 [View Article] [PubMed]
    [Google Scholar]
  17. Kazlauskas D, Varsani A, Koonin EV, Krupovic M. Multiple origins of prokaryotic and eukaryotic single-stranded DNA viruses from bacterial and archaeal plasmids. Nat Commun 2019; 10:3425 [View Article] [PubMed]
    [Google Scholar]
  18. Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinform 2014; 30:2114–2120 [View Article] [PubMed]
    [Google Scholar]
  19. Nurk S, Meleshko D, Korobeynikov A, Pevzner PA. metaSPAdes: a new versatile metagenomic assembler. Genome Res 2017; 27:824–834 [View Article] [PubMed]
    [Google Scholar]
  20. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. J Mol Biol 1990; 215:403–410 [View Article] [PubMed]
    [Google Scholar]
  21. Katoh K, Standley DM. MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Mol Biol Evol 2013; 30:772–780 [View Article] [PubMed]
    [Google Scholar]
  22. Guindon S, Dufayard J-F, Lefort V, Anisimova M, Hordijk W et al. New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Syst Biol 2010; 59:307–321 [View Article] [PubMed]
    [Google Scholar]
  23. Darriba D, Taboada GL, Doallo R, Posada D. ProtTest 3: fast selection of best-fit models of protein evolution. Bioinform 2011; 27:1164–1165 [View Article] [PubMed]
    [Google Scholar]
  24. Pei J, Grishin NV. PROMALS3D: multiple protein sequence alignment enhanced with evolutionary and three-dimensional structural information. Methods Mol Biol 2014; 1079:263–271 [View Article] [PubMed]
    [Google Scholar]
  25. Procter JB, Carstairs GM, Soares B, Mourão K, Ofoegbu TC et al. Alignment of biological sequences with jalview. Methods Mol Biol 2021; 2231:203–224 [View Article] [PubMed]
    [Google Scholar]
  26. Muhire BM, Varsani A, Martin DP. SDT: a virus classification tool based on pairwise sequence alignment and identity calculation. PLoS One 2014; 9:e108277 [View Article] [PubMed]
    [Google Scholar]
  27. Steinegger M, Meier M, Mirdita M, Vöhringer H, Haunsberger SJ et al. HH-suite3 for fast remote homology detection and deep protein annotation. BMC Bioinform 2019; 20:473 [View Article] [PubMed]
    [Google Scholar]
  28. Mirdita M, Schütze K, Moriwaki Y, Heo L, Ovchinnikov S et al. ColabFold: making protein folding accessible to all. Nat Methods 2022; 19:679–682 [View Article] [PubMed]
    [Google Scholar]
  29. Holm L. Dali server: structural unification of protein families. Nucleic Acids Res 2022; 50:W210–W215 [View Article] [PubMed]
    [Google Scholar]
  30. Meng EC, Goddard TD, Pettersen EF, Couch GS, Pearson ZJ et al. UCSF chimeraX: tools for structure building and analysis. Protein Sci 2023; 32:e4792 [View Article] [PubMed]
    [Google Scholar]
  31. McClatchie S, Juniper SK, Knox GA. Structure of a mudflat diatom community in the avon‐heathcote estuary, New Zealand. N Z J Mar Freshwater Res 1982; 16:299–309 [View Article]
    [Google Scholar]
  32. Chandler M, de la Cruz F, Dyda F, Hickman AB, Moncalian G et al. Breaking and joining single-stranded DNA: the HUH endonuclease superfamily. Nat Rev Microbiol 2013; 11:525–538 [View Article] [PubMed]
    [Google Scholar]
  33. Ilyina TV, Koonin EV. Conserved sequence motifs in the initiator proteins for rolling circle DNA replication encoded by diverse replicons from eubacteria, eucaryotes and archaebacteria. Nucleic Acids Res 1992; 20:3279–3285 [View Article] [PubMed]
    [Google Scholar]
  34. Koonin EV, Ilyina TV. Computer-assisted dissection of rolling circle DNA replication. Biosystems 1993; 30:241–268 [View Article] [PubMed]
    [Google Scholar]
  35. Gorbalenya AE, Koonin EV. Helicases: amino acid sequence comparisons and structure-function relationships. Curr Opin Struc Biol 1993; 3:419–429 [View Article]
    [Google Scholar]
  36. Gorbalenya AE, Koonin EV, Wolf YI. A new superfamily of putative NTP-binding domains encoded by genomes of small DNA and RNA viruses. FEBS Lett 1990; 262:145–148 [View Article] [PubMed]
    [Google Scholar]
  37. Krupovic M, Varsani A. Naryaviridae, Nenyaviridae, and Vilyaviridae: three new families of single-stranded DNA viruses in the phylum Cressdnaviricota. Arch Virol 2022; 167:2907–2921 [View Article] [PubMed]
    [Google Scholar]
  38. Varsani A, Harrach B, Roumagnac P, Benkő M, Breitbart M et al. 2024 taxonomy update for the family Circoviridae. Arch Virol 2024; 169:176 [View Article]
    [Google Scholar]
  39. Varsani A, Krupovic M. Sequence-based taxonomic framework for the classification of uncultured single-stranded DNA viruses of the family Genomoviridae. Virus Evol 2017; 3:vew037 [View Article] [PubMed]
    [Google Scholar]
  40. Varsani A, Krupovic M. Smacoviridae: a new family of animal-associated single-stranded DNA viruses. Arch Virol 2018; 163:2005–2015 [View Article] [PubMed]
    [Google Scholar]
  41. Siddell SG, Walker PJ, Lefkowitz EJ, Mushegian AR, Dutilh BE et al. Binomial nomenclature for virus species: a consultation. Arch Virol 2020; 165:519–525 [View Article] [PubMed]
    [Google Scholar]
  42. Lefeuvre P, Lett JM, Varsani A, Martin DP. Widely conserved recombination patterns among single-stranded DNA viruses. J Virol 2009; 83:2697–2707 [View Article] [PubMed]
    [Google Scholar]
  43. Martin DP, Biagini P, Lefeuvre P, Golden M, Roumagnac P et al. Recombination in eukaryotic single stranded DNA viruses. Viruses 2011; 3:1699–1738 [View Article] [PubMed]
    [Google Scholar]
  44. de la Higuera I, Kasun GW, Torrance EL, Pratt AA, Maluenda A et al. Unveiling crucivirus diversity by mining metagenomic data. mBio 2020; 11:e01410-20 [View Article] [PubMed]
    [Google Scholar]
  45. Kinsella CM, Bart A, Deijs M, Broekhuizen P, Kaczorowska J et al. Entamoeba and giardia parasites implicated as hosts of CRESS viruses. Nat Commun 2020; 11:4620 [View Article] [PubMed]
    [Google Scholar]
  46. Krupovic M, Zhi N, Li J, Hu G, Koonin EV et al. Multiple layers of chimerism in a single-stranded DNA virus discovered by deep sequencing. Genome Biol Evol 2015; 7:993–1001 [View Article] [PubMed]
    [Google Scholar]
  47. Roux S, Enault F, Bronner G, Vaulot D, Forterre P et al. Chimeric viruses blur the borders between the major groups of eukaryotic single-stranded DNA viruses. Nat Commun 2013; 4:2700 [View Article] [PubMed]
    [Google Scholar]
  48. Requião RD, Carneiro RL, Moreira MH, Ribeiro-Alves M, Rossetto S et al. Viruses with different genome types adopt a similar strategy to pack nucleic acids based on positively charged protein domains. Sci Rep 2020; 10:5470 [View Article] [PubMed]
    [Google Scholar]
  49. Sarker S, Terrón MC, Khandokar Y, Aragão D, Hardy JM et al. Structural insights into the assembly and regulation of distinct viral capsid complexes. Nat Commun 2016; 7:13014 [View Article] [PubMed]
    [Google Scholar]
  50. Butkovic A, Kraberger S, Smeele Z, Martin DP, Schmidlin K et al. Evolution of anelloviruses from a circovirus-like ancestor through gradual augmentation of the jelly-roll capsid protein. Virus Evol 2023; 9:vead035 [View Article] [PubMed]
    [Google Scholar]
  51. Liou S-H, Boggavarapu R, Cohen NR, Zhang Y, Sharma I et al. Structure of anellovirus-like particles reveal a mechanism for immune evasion. Nat Commun 2024; 15:7219 [View Article] [PubMed]
    [Google Scholar]
  52. De Koch MD, Krupovic M, Fielding R, Smith K, Schiavone K et al. Novel lineage of anelloviruses with large genomes identified in dolphins. J Virol 2025; 99:e0137024 [View Article] [PubMed]
    [Google Scholar]
  53. Varsani A, Hopkins A, Lund MC, Krupovic M. 2024 taxonomic update for the families Naryaviridae, Nenyaviridae, and Vilyaviridae. Arch Virol 2024; 170:18 [View Article] [PubMed]
    [Google Scholar]
  54. Richmond GS, Smith TK. Phospholipases A₁. Int J Mol Sci 2011; 12:588–612 [View Article] [PubMed]
    [Google Scholar]
  55. Bian C, Yuan C, Chen L, Meehan EJ, Jiang L et al. Crystal structure of a triacylglycerol lipase from Penicillium expansum at 1.3 a determined by sulfur SAD. Proteins 2010; 78:1601–1605 [View Article] [PubMed]
    [Google Scholar]
  56. Brzozowski AM, Savage H, Verma CS, Turkenburg JP, Lawson DM et al. Structural origins of the interfacial activation in Thermomyces (Humicola) lanuginosa Lipase. Biochemistry 2000; 39:15071–15082 [View Article]
    [Google Scholar]
  57. Lan D, Zhao G, Holzmann N, Yuan S, Wang J et al. Structure-guided rational design of a mono- and diacylglycerol lipase from Aspergillus oryzae : a single residue mutant increases the hydrolysis ability. J Agric Food Chem 2021; 69:5344–5352 [View Article]
    [Google Scholar]
  58. Cotmore SF, Agbandje-McKenna M, Canuti M, Chiorini JA, Eis-Hubinger A-M et al. ICTV virus taxonomy profile: Parvoviridae. J Gen Virol 2019; 100:367–368 [View Article] [PubMed]
    [Google Scholar]
  59. Cotmore SF, Tattersall P. Parvoviral host range and cell entry mechanisms. Adv Virus Res 2007; 70:183–232 [View Article] [PubMed]
    [Google Scholar]
  60. Farr GA, Zhang L, Tattersall P. Parvoviral virions deploy a capsid-tethered lipolytic enzyme to breach the endosomal membrane during cell entry. Proc Natl Acad Sci USA 2005; 102:17148–17153 [View Article] [PubMed]
    [Google Scholar]
  61. Wolf YI, Silas S, Wang Y, Wu S, Bocek M et al. Doubling of the known set of RNA viruses by metagenomic analysis of an aquatic virome. Nat Microbiol 2020; 5:1262–1270 [View Article] [PubMed]
    [Google Scholar]
  62. Koonin EV, Fischer MG, Kuhn JH, Krupovic M. The polinton-like supergroup of viruses: evolution, molecular biology, and taxonomy. Microbiol Mol Biol Rev 2024; 88:e0008623 [View Article] [PubMed]
    [Google Scholar]
  63. Yutin N, Shevchenko S, Kapitonov V, Krupovic M, Koonin EV. A novel group of diverse Polinton-like viruses discovered by metagenome analysis. BMC Biol 2015; 13:95 [View Article] [PubMed]
    [Google Scholar]
  64. Born D, Reuter L, Mersdorf U, Mueller M, Fischer MG et al. Capsid protein structure, self-assembly, and processing reveal morphogenesis of the marine virophage mavirus. Proc Natl Acad Sci USA 2018; 115:7332–7337 [View Article] [PubMed]
    [Google Scholar]
  65. Edwards BR, Thamatrakoln K, Fredricks HF, Bidle KD, Van Mooy BAS. Viral infection leads to a unique suite of allelopathic chemical signals in three diatom host-virus pairs. Mar Drugs 2024; 22:228 [View Article] [PubMed]
    [Google Scholar]
  66. Zerbini FM, Siddell SG, Mushegian AR, Walker PJ, Lefkowitz EJ et al. Differentiating between viruses and virus species by writing their names correctly. Arch Virol 2022; 167:1231–1234 [View Article] [PubMed]
    [Google Scholar]
  67. Gilchrist CLM, Chooi YH. clinker & clustermap.js: automatic generation of gene cluster comparison figures. Bioinform 2021; 37:2473–2475 [View Article] [PubMed]
    [Google Scholar]
  68. Crooks GE, Hon G, Chandonia JM, Brenner SE. WebLogo: a sequence logo generator. Genome Res 2004; 14:1188–1190 [View Article] [PubMed]
    [Google Scholar]
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