PDF
Abstract
Aim: Temperate phages are known to heavily impact the growth of their host, be it in a positive way, e.g., when beneficial genes are provided by the phage, or negatively when lysis occurs after prophage induction. This study provides an in-depth look into the distribution and variety of prophages in Latilactobacillus curvatus (L. curvatus). This species is found in a wide variety of ecological niches and is routinely used as a meat starter culture.
Methods: Fourty five L. curvatus genomes were screened for prophages. The intact predicted prophages and their chromosomal integration loci were described. Six L. curvatus lysogens were analysed for phage-mediated lysis post induction via UV light and/or mitomycin C. Their lysates were analysed for phage particles via viral DNA sequencing and transmission electron microscopy.
Results: Two hundred and six prophage sequences of any completeness were detected within L. curvatus genomes. The 50 as intact predicted prophages show high levels of genetic diversity on an intraspecies level with conserved regions mostly in the replication and head/tail gene clusters. Twelve chromosomal loci, mostly tRNA genes, were identified, where intact L. curvatus phages were integrated. The six analysed L. curvatus lysogens showed strain-dependent lysis in various degrees after induction, yet only four of their lysates appeared to contain fully assembled virions with the siphovirus morphotype.
Conclusion: Our data demonstrate that L. curvatus is a (pro)phage-susceptible species, harbouring multiple intact prophages and remnant sequences thereof. This knowledge provides a basis to study phage-host interaction influencing microbial communities in food fermentations.
Keywords
Latilactobacillus curvatus
/
temperate bacteriophages
/
prophages
/
induction
/
lysis
Cite this article
Download citation ▾
Conrad L. Ambros, Matthias A. Ehrmann.
Distribution, inducibility, and characteristics of Latilactobacillus curvatus temperate phages.
Microbiome Research Reports, 2023, 2(4): 34 DOI:10.20517/mrr.2023.18
| [1] |
Samson JE.Bacteriophages in food fermentations: new frontiers in a continuous arms race.Annu Rev Food Sci Technol2013;4:347-68
|
| [2] |
Komora N,Ferreira V.The protective effect of food matrices on Listeria lytic bacteriophage P100 application towards high pressure processing.Food Microbiol2018;76:416-25
|
| [3] |
Shebs-Maurine EL,Laidler ST.Effects of bacteriophages and peroxyacetic acid applications on beef contaminated with Salmonella during different grinding stages.Meat Sci2021;173:108407
|
| [4] |
Merabishvili M,Verbeken G.Quality-controlled small-scale production of a well-defined bacteriophage cocktail for use in human clinical trials.PLoS One2009;4:e4944 PMCID:PMC2654153
|
| [5] |
Loessner MJ.Bacteriophage endolysins - current state of research and applications.Curr Opin Microbiol2005;8:480-7
|
| [6] |
Lunde M,Blatny JM.Effects of diverse environmental conditions on ϕ LC3 prophage stability in Lactococcus lactis.Appl Environ Microbiol2005;71:721-7 PMCID:PMC546743
|
| [7] |
Feyereisen M,Neve H.Biodiversity and classification of phages infecting Lactobacillus brevis.Front Microbiol2019;10:2396 PMCID:PMC6805780
|
| [8] |
Oh JH,Zhang S.Prophages in Lactobacillus reuteri are associated with fitness trade-offs but can increase competitiveness in the gut ecosystem.Appl Environ Microbiol2019;86:e01922-19 PMCID:PMC6912086
|
| [9] |
Wendling CC,Hall AR.Fitness benefits to bacteria of carrying prophages and prophage-encoded antibiotic-resistance genes peak in different environments.Evolution2021;75:515-28 PMCID:PMC7986917
|
| [10] |
Brown EM,Temple ER.Gut microbiome ADP-ribosyltransferases are widespread phage-encoded fitness factors.Cell Host Microbe2021;29:1351-65.e11 PMCID:PMC8429246
|
| [11] |
Liu Y,Boeren S,Smid EJ.Extracellular vesicle formation in Lactococcus lactis is stimulated by prophage-encoded holin-lysin system.Microb Biotechnol2022;15:1281-95 PMCID:PMC8966010
|
| [12] |
Briggiler Marcó M, Garneau JE, Tremblay D, Quiberoni A, Moineau S. Characterization of two virulent phages of Lactobacillus plantarum.Appl Environ Microbiol2012;78:8719-34 PMCID:PMC3502894
|
| [13] |
Ehrmann MA,Picozzi C,Vogel RF.The genome of the Lactobacillus sanfranciscensis temperate phage EV3.BMC Res Notes2013;6:514 PMCID:PMC4234937
|
| [14] |
Pei Z,Han X.Comprehensive scanning of prophages in Lactobacillus: distribution, diversity, antibiotic resistance genes, and linkages with CRISPR-Cas systems.mSystems2021;6:e0121120
|
| [15] |
Leroy F,De Vuyst L.Functional meat starter cultures for improved sausage fermentation.Int J Food Microbiol2006;106:270-85
|
| [16] |
Bassi D,Belloso Daza MV.Taxonomical identification and safety characterization of lactobacillaceae from mediterranean natural fermented sausages.Foods2022;11:2776 PMCID:PMC9497648
|
| [17] |
Jo SG,Lee JY.Lactobacillus curvatus WiKim38 isolated from kimchi induces IL-10 production in dendritic cells and alleviates DSS-induced colitis in mice.J Microbiol2016;54:503-9
|
| [18] |
Baek HW,Lee YG.Dynamic interactions of lactic acid bacteria in Korean sourdough during back-slopping process.J Appl Microbiol2021;131:2325-35
|
| [19] |
Sánchez I,Poveda JM,Palop L.Phenotypic and genotypic characterization of lactobacilli isolated from Spanish goat cheeses.Int J Food Microbiol2005;102:355-62
|
| [20] |
Bulgasem BY,Hassan Z,Fnaish SG.Antifungal activity of lactic acid bacteria strains isolated from natural honey against pathogenic candida species.Mycobiology2016;44:302-9 PMCID:PMC5287163
|
| [21] |
Ambros CL.Distribution, inducibility, and characterisation of prophages in Latilactobacillus sakei.BMC Microbiol2022;22:267 PMCID:PMC9641780
|
| [22] |
Mo SJ,Hong HJ.Effects of Lactobacillus curvatus HY7601 and Lactobacillus plantarum KY1032 on overweight and the gut microbiota in humans: randomized, double-blinded, placebo-controlled clinical trial.Nutrients2022;14:2484 PMCID:PMC9228474
|
| [23] |
Zhang Y,Zhao MD,Choi SH.Screening and identification of Latilactobacillus curvatus Z12 from rumen fluid of an adult female sika deer as a potential probiotic for feed additives.Front Vet Sci2021;8:753527 PMCID:PMC8566888
|
| [24] |
Koch C,Lurz R,Beutin L.Isolation of a lysogenic bacteriophage carrying the stx1OX3 gene, which is closely associated with Shiga toxin-producing Escherichia coli strains from sheep and humans.J Clin Microbiol2001;39:3992-8 PMCID:PMC88477
|
| [25] |
Townsend EM,Muscatt G.The human gut phageome: origins and roles in the human gut microbiome.Front Cell Infect Microbiol2021;11:643214 PMCID:PMC8213399
|
| [26] |
DSMZ. MRS medium.doc. 2007. Available from: https://www.dsmz.de/microorganisms/medium/pdf/DSMZ_Medium11.pdf. [Last accessed on 5 Sep 2023]
|
| [27] |
Cold Spring Harbor Protocols. SM buffer with gelatin. Available from: https://cshprotocols.cshlp.org/content/2006/1/pdb.rec466.full?text-only=true. [Last accessed on 5 Sep 2023]
|
| [28] |
Kaletta J,Griebler C,Kurmayer R.A rigorous assessment and comparison of enumeration methods for environmental viruses.Sci Rep2020;10:18625 PMCID:PMC7596560
|
| [29] |
Wick RR,Gorrie CL.Unicycler: resolving bacterial genome assemblies from short and long sequencing reads.PLoS Comput Biol2017;13:e1005595 PMCID:PMC5481147
|
| [30] |
Li W,Haft DH.RefSeq: expanding the prokaryotic genome annotation pipeline reach with protein family model curation.Nucleic Acids Res2021;49:D1020-8 PMCID:PMC7779008
|
| [31] |
Haft DH,Badretdin A.RefSeq: an update on prokaryotic genome annotation and curation.Nucleic Acids Res2018;46:D851-60 PMCID:PMC5753331
|
| [32] |
Tatusova T,Badretdin A.NCBI prokaryotic genome annotation pipeline.Nucleic Acids Res2016;44:6614-24 PMCID:PMC5001611
|
| [33] |
SM buffer.Cold Spring Harb Protoc2006;2006:pdb.rec8111
|
| [34] |
Protocol for phage DNA extraction with Phenol:chloroform. Available from: https://cpt.tamu.edu/wordpress/wp-content/uploads/2018/09/Phage-DNA-Extraction-by-PhenolChloroform-Protocol.pdf. [Last accessed on 5 Sep 2023]
|
| [35] |
Richter M,Oliver Glöckner F.JSpeciesWS: a web server for prokaryotic species circumscription based on pairwise genome comparison.Bioinformatics2016;32:929-31 PMCID:PMC5939971
|
| [36] |
Arndt D,Marcu A.PHASTER: a better, faster version of the PHAST phage search tool.Nucleic Acids Res2016;44:W16-21 PMCID:PMC4987931
|
| [37] |
Zhou Y,Lynch KH,Wishart DS.PHAST: a fast phage search tool.Nucleic Acids Res2011;39:W347-52 PMCID:PMC3125810
|
| [38] |
Lowe TM.tRNAscan-SE On-line: integrating search and context for analysis of transfer RNA genes.Nucleic Acids Res2016;44:W54-7 PMCID:PMC4987944
|
| [39] |
Chan PP,Mak AJ.tRNAscan-SE 2.0: improved detection and functional classification of transfer RNA genes.Nucleic Acids Res2021;49:9077-96 PMCID:PMC8450103
|
| [40] |
Roberts RJ,Posfai J.REBASE: a database for DNA restriction and modification: enzymes, genes and genomes.Nucleic Acids Res2023;51:D629-30 PMCID:PMC9825431
|
| [41] |
ICTV. Taxon names are written differently from virus names. Available from: https://talk.ictvonline.org/information/w/faq/386/how-to-write-virus-species-and-other-taxa-names. [Last accessed on 5 Sep 2023]
|
| [42] |
Sullivan MJ,Beatson SA.Easyfig: a genome comparison visualizer.Bioinformatics2011;27:1009-10 PMCID:PMC3065679
|
| [43] |
Kumar S,Li M,Tamura K.MEGA X: molecular evolutionary genetics analysis across computing platforms.Mol Biol Evol2018;35:1547-9 PMCID:PMC5967553
|
| [44] |
Lee J,Kim B,Jeong G.Identification of Lactobacillus sakei and Lactobacillus curvatus by multiplex PCR-based restriction enzyme analysis.J Microbiol Methods2004;59:1-6
|
| [45] |
Williams KP.Integration sites for genetic elements in prokaryotic tRNA and tmRNA genes: sublocation preference of integrase subfamilies.Nucleic Acids Res2002;30:866-75 PMCID:PMC100330
|
| [46] |
Qin Y,Vesper O.The highly conserved LepA is a ribosomal elongation factor that back-translocates the ribosome.Cell2006;127:721-33
|
| [47] |
Bobay LM,Touchon M.The adaptation of temperate bacteriophages to their host genomes.Mol Biol Evol2013;30:737-51 PMCID:PMC3603311
|
| [48] |
Hotopp JCD,Kumar N.Comparative genomics of Neisseria meningitidis: core genome, islands of horizontal transfer and pathogen-specific genes.Microbiology2006;152:3733-49
|
| [49] |
Bobay LM,Rocha EP.Pervasive domestication of defective prophages by bacteria.Proc Natl Acad Sci U S A2014;111:12127-32 PMCID:PMC4143005
|
| [50] |
Yang JY,Miranda-Sanchez F.Degradation of host translational machinery drives tRNA acquisition in viruses.Cell Syst2021;12:771-9.e5 PMCID:PMC8826309
|
| [51] |
Bailly-Bechet M,Rocha E.Causes for the intriguing presence of tRNAs in phages.Genome Res2007;17:1486-95 PMCID:PMC1987346
|
| [52] |
Yoshikawa G,Blanc-Mathieu R.Xanthomonas citri jumbo phage XacN1 exhibits a wide host range and high complement of tRNA genes.Sci Rep2018;8:4486 PMCID:PMC5852040
|
| [53] |
Tomikawa C,Guérineau V.Characterization of redundant tRNAIles with CAU and UAU anticodons in Lactobacillus plantarum.J Biochem2018;163:233-41
|
| [54] |
McDonald MJ,Swamy KB,Leu JY.The evolutionary dynamics of tRNA-gene copy number and codon-use in E. coli.BMC Evol Biol2015;15:163 PMCID:PMC4539685
|
| [55] |
Murphy J,Ainsworth S,van Sinderen D.Bacteriophage orphan DNA methyltransferases: insights from their bacterial origin, function, and occurrence.Appl Environ Microbiol2013;79:7547-55 PMCID:PMC3837797
|
| [56] |
Pingoud A,Pingoud V.Type II restriction endonucleases: structure and mechanism.Cell Mol Life Sci2005;62:685-707
|
| [57] |
Bickle TA.Biology of DNA restriction.Microbiol Rev1993;57:434-50 PMCID:PMC372918
|
| [58] |
Sternberg N.Cleavage of the bacteriophage P1 packaging site (pac) is regulated by adenine methylation.Proc Natl Acad Sci U S A1990;87:8070-4 PMCID:PMC54894
|
| [59] |
Jensen TØ,Redl S.Genome-wide systematic identification of methyltransferase recognition and modification patterns.Nat Commun2019;10:3311 PMCID:PMC6700114
|
| [60] |
Marinus MG.Biological function for 6-methyladenine residues in the DNA of Escherichia coli K12.J Mol Biol1974;85:309-22
|
| [61] |
Lahue RS,Modrich P.DNA mismatch correction in a defined system.Science1989;245:160-4
|
| [62] |
Smith MJ.DNA methylation in lysogens of pathogenic Burkholderia spp. Requires prophage induction and is restricted to excised phage DNA.J Bacteriol2005;187:1196-200 PMCID:PMC545696
|
| [63] |
Kenyon CJ.DNA-damaging agents stimulate gene expression at specific loci in Escherichia coli.Proc Natl Acad Sci U S A1980;77:2819-23 PMCID:PMC349496
|