An alternative molecular marker with respect to the 16S rRNA gene demonstrating better identification and phylogenetic parameters has not been designed for the whole Bifidobacteriaceae family, which includes the genus Bifidobacterium and scardovial genera. Therefore, the aim of the study was to find such a gene in available genomic sequences, suggest appropriate means and conditions for asmplification and sequencing of the desired region of the selected gene in various strains of the bacterial family and verify the importance in classification and phylogeny. Specific primers flanking the variable region (~800 pb) within the pyrG gene encoding the CTP synthetase were designed by means of gene sequences retrieved from the genomes of strains belonging to the family Bifidobacteriaceae. The functionality and specificity of the primers were subsequently tested on the wild (7) and type strains of bifidobacteria (36) and scardovia (7). Comparative and phylogenetic studies based on obtained sequences revealed actual significance in classification and phylogeny of the Bifidobacteriaceae family. Gene statistics (percentages of mean sequence similarities and identical sites, mean number of nucleotide differences, P- and K-distances) and phylogenetic analyses (congruence between tree topologies, percentages of bootstrap values >50 and 70%) indicate that the pyrG gene represents an alternative identification and phylogenetic marker exhibiting higher discriminatory power among strains, (sub)species, and genera than the 16S rRNA gene. Sequences of the particular gene fragment, simply achieved through specific primers, enable more precisely to classify and evaluate phylogeny of the family Bifidobacteriaceae including, with some exceptions, health-promoting probiotic bacteria.

Faculty of Agrobiology Food and Natural Resources Department of Microbiology Nutrition and Dietetics Czech University of Life Sciences Prague 6 Suchdol Czechia

Institute of Animal Physiology and Genetics of the Czech Academy of Sciences Prague 4 Krč Czechia

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Abayneh, T. , Colquhoun, D. J. , & Sørum, H. (2012). Multi‐locus Sequence Analysis (MLSA) of PubMed DOI

Adékambi, T. , Butler, R. W. , Hanrahan, F. , Delcher, A. L. , Drancourt, M. , & Shinnick, T. M. (2011). Core gene set as the basis of multilocus sequence analysis of the subclass Actinobacteridae. PLoS ONE, 6, e14792 10.1371/journal.pone.0014792 PubMed DOI PMC

Avcin, S. L. , Pokorn, M. , Kitanovski, L. , Premru, M. M. , & Jazbec, J. (2015). PubMed DOI PMC

Barberis, C. M. , Cittadini, R. M. , Almuzara, M. N. , Feinsilberg, A. , Famiglietti, A. M. , Ramírez, M. S. , & Vay, C. A. (2012). Recurrent urinary infection with PubMed DOI PMC

Biavati, B. , & Mattarelli, P. (2012). Genus

Bunesova, V. , Vlkova, E. , Rada, V. , Killer, J. , & Musilova, S. (2014). Bifidobacteria from the gastrointestinal tract of animals: Differences and similarities. Beneficial Microbes, 5, 377–388. 10.3920/BM2013.0081 PubMed DOI

Castresana, J. (2000). Selection of conserved blocks from multiple alignments for their use in phylogenetic analysis. Molecular Biology and Evolution, 17, 540–552. 10.1093/oxfordjournals.molbev.a026334 PubMed DOI

Delcenserie, V. , Gavini, F. , Beerens, H. , Tresse, O. , Franssen, C. , & Daube, G. (2007). Description of a new species, PubMed DOI

Delétoile, A. , Passet, V. , Aires, J. , Chambaud, I. , Butel, M. J. , Smokvina, T. , & Brisse, S. (2010). Species delineation and clonal diversity in four PubMed DOI

Diancourt, L. , Passet, V. , Chervaux, C. , Garault, P. , Smokvina, T. , & Brisse, S. (2007). Multilocus sequence typing of PubMed DOI PMC

Dong, X. , Xin, Y. , Jian, W. , Liu, X. , & Ling, D. (2000). PubMed DOI

Downes, J. , Mantzourani, M. , Beighton, D. , Hooper, S. , Wilson, M. J. , Nicholson, A. , & Wade, W. G. (2011). PubMed DOI

Fournier, P. E. , Suhre, K. , Fournous, G. , & Raoult, D. (2006). Estimation of prokaryote genomic DNA G+C content by sequencing universally conserved genes. International Journal of Systematic and Evolutionary Microbiology, 56, 1025–1029. 10.1099/ijs.0.63903-0 PubMed DOI

Glaeser, S. P. , & Kämpfer, P. (2015). Multilocus sequence analysis (MLSA) in prokaryotic taxonomy. Systematic and Applied Microbiology, 38, 237–245. 10.1016/j.syapm.2015.03.007 PubMed DOI

Gupta, R. S. , Nanda, A. , & Khadka, B. (2017). Novel molecular, structural and evolutionary characteristics of the phosphoketolases from bifidobacteria and Coriobacteriales. PLoS ONE, 12, e0172176 10.1371/journal.pone.0172176 PubMed DOI PMC

Huys, G. , Vancanneyt, M. , D′Haene, K. , Falsen, E. , Wauters, G. , & Vandamme, P. (2007). PubMed DOI

Jian, W. , Zhu, L. , & Dong, X. (2001). New approach to phylogenetic analysis of the genus PubMed DOI

Killer, J. , Havlik, J. , Bunesova, V. , Vlkova, E. , & Benada, O. (2014). PubMed DOI

Killer, J. , Kopečný, J. , Mrázek, J. , Havlík, J. , Koppová, I. , Benada, O. , … Kofroňová, O. (2010). PubMed DOI

Killer, J. , Ročková, Š. , Vlková, E. , Rada, V. , Havlík, J. , Kopečný, J. , … Profousová, I. (2013a). PubMed DOI

Killer, J. , Sedláček, I. , Rada, V. , Havlík, J. , & Kopečný, J. (2013b). Reclassification of PubMed DOI

Librado, P. , & Rozas, J. (2009). DnaSP v5: A software for comprehensive analysis of DNA polymorphism data. Bioinformatics, 25, 1451–1452. 10.1093/bioinformatics/btp187 PubMed DOI

Lugli, G. A. , Milani, C. , Turroni, F. , Duranti, S. , Ferrario, C. , Viappiani, A. , … Ventura, M. (2014). Investigation of the evolutionary development of the genus PubMed DOI PMC

Lugli, G. A. , Milani, C. , Turroni, F. , Duranti, S. , Mancabelli, L. , Mangifesta, M. , … Ventura, M. (2017). Comparative genomic and phylogenomic analyses of the PubMed DOI PMC

Mahlen, S. D. , & Clarridge, J. E. (2009). Site and clinical significance of PubMed DOI PMC

Martin, D. P. , Lemey, P. , Lott, M. , Moulton, V. , Posada, D. , & Lefeuvre, P. (2010). RDP3: A flexible and fast computer program for analyzing recombination. Bioinformatics, 26, 2462–2463. 10.1093/bioinformatics/btq467 PubMed DOI PMC

Martin, D. , & Rybicki, E. (2000). RDP: Detection of recombination amongst aligned sequences. Bioinformatics, 16, 562–563. 10.1093/bioinformatics/16.6.562 PubMed DOI

Mattarelli, P. , Holzapfel, W. , Franz, C. M. , Endo, A. , Felis, G. E. , Hammes, W. , … Dellaglio, F. (2014). Recommended minimal standards for description of new taxa of the genera PubMed DOI

Milani, C. , Lugli, G. A. , Duranti, S. , Turroni, F. , Bottacini, F. , Mangifesta, M. , … Ventura, M. (2014). Genomic encyclopedia of type strains of the genus PubMed DOI PMC

Milani, C. , Turroni, F. , Duranti, S. , Lugli, G. A. , Mancabelli, L. , Ferrario, C. , … Ventura, M. (2015). Genomics of the genus PubMed PMC

Naushad, H. S. , & Gupta, R. S. (2013). Phylogenomics and molecular signatures for species from the plant pathogen‐containing order xanthomonadales. PLoS ONE, 8, e55216 10.1371/journal.pone.0055216 PubMed DOI PMC

Nuñez, H. , Loyola, D. , Cárdenas, J. P. , Holmes, D. S. , Johnson, D. B. , & Quatrini, R. (2014). Multi locus sequence typing scheme for PubMed DOI

O'Callaghan, A. , & van Sinderen, D. (2016). Bifidobacteria and their role as members of the human gut microbiota. Frontiers in Microbiology, 7, 925. PubMed PMC

Pechar, R. , Killer, J. , Mekadim, C. , Geigerová, M. , & Rada, V. (2017a). Classification of culturable bifidobacterial population from colonic samples of wild pigs ( PubMed DOI

Pechar, R. , Killer, J. , Salmonová, H. , Geigerová, M. , Švejstil, R. , Švec, P. , … Benada, O. (2017b). PubMed

Pechar, R. , Killer, J. , Švejstil, R. , Salmonová, H. , Geigerová, M. , Bunešová, V. , … Benada, O. (2017c). PubMed

Rada, V. , & Petr, J. (2000). A new selective medium for the isolation of glucose non‐fermenting bifidobacteria from hen caeca. Journal of Microbiological Methods, 43, 127–132. 10.1016/S0167-7012(00)00205-0 PubMed DOI

Roux, S. , Enault, F. , Bronner, G. , & Debroas, D. (2011). Comparison of 16S rRNA and protein‐coding genes as molecular markers for assessing microbial diversity (Bacteria and Archaea) in ecosystems. FEMS Microbiology Ecology, 78, 617–628. 10.1111/j.1574-6941.2011.01190.x PubMed DOI

Rudenko, N. , Golovchenko, M. , Belfiore, N. M. , Grubhoffer, L. , & Oliver, J. H. Jr (2014). Divergence of PubMed DOI PMC

Sarmiento‐Rubiano, L. A. , Berger, B. , Moine, D. , Zúñiga, M. , Pérez‐Martínez, G. , & Yebra, M. J. (2010). Characterization of a novel PubMed DOI PMC

Satokari, R. M. , Vaughan, E. E. , Akkermans, A. D. , Saarela, M. , & de Vos, W. M. (2001). Bifidobacterial diversity in human feces detected by genus‐specific PCR and denaturing gradient gel electrophoresis. Applied and Environmental Microbiology, 67, 504–513. 10.1128/AEM.67.2.504-513.2001 PubMed DOI PMC

Scardovi, V. (1986). Genus

Scardovi, V. , Trovatelli, L. D. , Biavati, B. , & Zani, G. (1979). DOI

Sechovcová, H. , Killer, J. , Pechar, R. , Geigerová, M. , Švejstil, R. , Salmonová, H. , … Benada, O. (2017). PubMed

Simpson, P. J. , Ross, R. P. , Fitzgerald, G. F. , & Stanton, C. (2004). PubMed DOI

Sun, Z. , Zhang, W. , Guo, C. , Yang, X. , Liu, W. , Wu, Y. , … Zhang, H. (2015). Comparative genomic analysis of 45 type strains of the genus PubMed DOI PMC

Tamura, K. , Peterson, D. , Peterson, N. , Stecher, G. , Nei, M. , & Kumar, S. (2011). MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution, 28, 2731–2739. 10.1093/molbev/msr121 PubMed DOI PMC

Tanner, A. C. , Mathney, J. M. , Kent, R. L. , Chalmers, N. I. , Hughes, C. V. , Loo, C. Y. , … Dewhirst, F. E. (2011). Cultivable anaerobic microbiota of severe early childhood caries. Journal of Clinical Microbiology, 49, 1464–1474. 10.1128/JCM.02427-10 PubMed DOI PMC

Tomasini, N. , Lauthier, J. J. , Llewellyn, M. S. , & Diosque, P. (2013). MLSTest: Novel software for multi‐locus sequence data analysis in eukaryotic organisms. Infection, Genetics and Evolution, 20, 188–196. 10.1016/j.meegid.2013.08.029 PubMed DOI

Untergasser, A. , Cutcutache, I. , Koressaar, T. , Ye, J. , Faircloth, B. C. , Remm, M. , & Rozen, S. G. (2012). Primer3‐new capabilities and interfaces. Nucleic Acids Research, 40, e115 10.1093/nar/gks596 PubMed DOI PMC

Valdez, R. M. , Dos Santos, V. R. , Caiaffa, K. S. , Danelon, M. , Arthur, R. A. , Negrini, T. C. , … Duque, C. (2016). Comparative in vitro investigation of the cariogenic potential of bifidobacteria. Archives of Oral Biology, 71, 97–103. 10.1016/j.archoralbio.2016.07.005 PubMed DOI

Ventura, M. , Canchaya, C. , Del Casale, A. , Dellaglio, F. , Neviani, E. , Fitzgerald, G. F. , & van Sinderen, D. (2006). Analysis of bifidobacterial evolution using a multilocus approach. International Journal of Systematic and Evolutionary Microbiology, 56, 2783–2792. 10.1099/ijs.0.64233-0 PubMed DOI

Větrovský, T. , & Baldrian, P. (2013). The variability of the 16S rRNA gene in bacterial genomes and its consequences for bacterial community analyses. PLoS ONE, 8, e57923 10.1371/journal.pone.0057923 PubMed DOI PMC

Weber, E. , Reynaud, Q. , Suy, F. , Gagneux‐Brunon, A. , Carricajo, A. , Guillot, A. , & Botelho‐Nevers, E. (2015). PubMed DOI

Wu, D. , Jospin, G. , & Eisen, J. A. (2013). Systematic identification of gene families for use as “markers” for phylogenetic and phylogeny‐driven ecological studies of bacteria and archaea and their major subgroups. PLoS ONE, 8, e77033 10.1371/journal.pone.0077033 PubMed DOI PMC

Yoon, S. H. , Ha, S. M. , Kwon, S. , Lim, J. , Kim, Y. , Seo, H. , & Chun, J. (2017). Introducing EzBioCloud: A taxonomically united database of 16S rRNA gene sequences and whole‐genome assemblies. International Journal of Systematic and Evolutionary Microbiology, 67, 1613–1617. PubMed PMC

Zhang, G. , Gao, B. , Adeolu, M. , Khadka, B. , & Gupta, R. S. (2016). Phylogenomic analyses and comparative studies on genomes of the bifidobacteriales: Identification of molecular signatures specific for the order bifidobacteriales and its different subclades. Frontiers in Microbiology, 7, 978. PubMed PMC

Selection of Probiotics for Honey Bees: The In Vitro Inhibition of Paenibacillus larvae, Melissococcus plutonius, and Serratia marcescens Strain Sicaria by Host-Specific Lactobacilli and Bifidobacteria

The bifidobacterial distribution in the microbiome of captive primates reflects parvorder and feed specialization of the host

Glutamine synthetase type I (glnAI) represents a rewarding molecular marker in the classification of bifidobacteria and related genera

Fragment of the aspartyl-tRNA synthetase applicable as a shared classification and phylogenetic marker in particular representatives of the order Lactobacillales

The threonine-tRNA ligase gene region is applicable in classification, typing, and phylogenetic analysis of bifidobacteria

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GENBANK
NR042759, KT351222, AM419460, KT351223, NR041347, KT351224, KT351225, NR118047, KT351226, HQ842704, KT351227, NR112093, KT351228, NC008618, KY365559, KY705017, KY746715, KT351229, KT351230, CP001606, NC012815, LC071851, KT351231, KT351232, KY365560, KY705018, KY746716, KT351233, NR104872, KT351234, KT351235, AB006658, KT351236, KY705019, KY746717, KT351237, KY705020, KY746718, KT351238, KT351239, AP012326, KT351240, KT351241, KT351242, NR043439, KT351243, NR112779, KT351244, AP010889, KT351245, AP010888, KT351246, KT351247, KY705021, KY746719, KT351248, KT351249, KT351250, KY705022, KY746720, KT351251, KT351252, KT351253, KT351254, KT351255, KT351256, AY148470, KT351257, KY705023, KY746721, KT351258, KT351259, KT351260, AP009256, AB042288