1887

Abstract

A Gram-stain-positive, endospore-forming endophytic bacterial strain designated MHSD28 was isolated from surface-sterilized leaves of collected from Eisleben, Botlokwa, Limpopo Province, South Africa. The phenotypic and phylogenetic characteristics of strain MHSD28 were consistent with those of members within the group. Comparative analysis between this strain and its relatives confirmed that it belongs to this group and forms a monophyletic branch. The digital DNA–DNA hybridization values between strain MHSD28 and its relatives were lower than the 70 % threshold for species delineation. To further determine its phylogenetic position, multi-locus sequence analysis (MLSA) based on five concatenated housekeeping gene (, , , and ) sequences, phenotypic analysis, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) biotyper identification, fatty acid and polar lipid profile analyses were carried out. Phenotypic characterization, MLSA, whole genome sequence based analyses and MALDI-TOF results placed strain MHSD28 within the group. The major fatty acids were iso-C and summed feature 3 and the main polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The respiratory quinone was menaquinone-7. The cell-wall peptidoglycan structure included -diaminopimelic acid. Considering the above results, strain MHSD28 represents a novel species of the group, for which the name sp. nov. is proposed. The type strain is MHSD28 (=BD 2262=LMG 32287=CECT 30671).

Funding
This study was supported by the:
  • ZAR-NRF (Award Thuthuka grant no TTK170405225920)
    • Principle Award Recipient: MahloroHope Serepa-Dlamini
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
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2023-10-25
2024-05-09
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References

  1. Ceuppens S, Boon N, Uyttendaele M. Diversity of Bacillus cereus group strains is reflected in their broad range of pathogenicity and diverse ecological lifestyles. FEMS Microbiol Ecol 2013; 84:433–450 [View Article] [PubMed]
    [Google Scholar]
  2. Smith NR, Gordon RE, Clark FE. Aerobic spore-forming bacteria: US government printing office; 1952
  3. Logan NA, Carman JA, Melling J, Berkeley RC. Identification of Bacillus anthracis by API tests. J Med Microbiol 1985; 20:75–85 [View Article] [PubMed]
    [Google Scholar]
  4. Nakamura LK. DNA relatedness among Bacillus thuringiensis serovars. Int J Syst Bacteriol 1994; 44:125–129 [View Article] [PubMed]
    [Google Scholar]
  5. Nakamura LK, Jackson MA. Clarification of the taxonomy of Bacillus mycoides. Int J Syst Bacteriol 1995; 45:46–49 [View Article] [PubMed]
    [Google Scholar]
  6. Nakamura LK. Bacillus pseudomycoides sp. nov. Int J Syst Bacteriol 1998; 48:1031–1035 [View Article]
    [Google Scholar]
  7. Lechner S, Mayr R, Francis KP, Prüss BM, Kaplan T et al. Bacillus weihenstephanensis sp. nov. is a new psychrotolerant species of the Bacillus cereus group. Int J Syst Bacteriol 1998; 48 Pt 4:1373–1382 [View Article] [PubMed]
    [Google Scholar]
  8. Guinebretière M-H, Auger S, Galleron N, Contzen M, De Sarrau B et al. Bacillus cytotoxicus sp. nov. is a novel thermotolerant species of the Bacillus cereus group occasionally associated with food poisoning. Int J Syst Evol Microbiol 2013; 63:31–40 [View Article] [PubMed]
    [Google Scholar]
  9. Miller RA, Beno SM, Kent DJ, Carroll LM, Martin NH et al. Bacillus wiedmannii sp. nov., a psychrotolerant and cytotoxic Bacillus cereus group species isolated from dairy foods and dairy environments. Int J Syst Evol Microbiol 2016; 66:4744–4753 [View Article] [PubMed]
    [Google Scholar]
  10. Jiménez G, Urdiain M, Cifuentes A, López-López A, Blanch AR et al. Description of Bacillus toyonensis sp. nov., a novel species of the Bacillus cereus group, and pairwise genome comparisons of the species of the group by means of ANI calculations. Syst Appl Microbiol 2013; 36:383–391 [View Article] [PubMed]
    [Google Scholar]
  11. Liu Y, Du J, Lai Q, Zeng R, Ye D et al. Proposal of nine novel species of the Bacillus cereus group. Int J Syst Evol Microbiol 2017; 67:2499–2508 [View Article]
    [Google Scholar]
  12. Liu X, Wang L, Han M, Xue Q, Zhang G et al. Bacillus fungorum sp. nov., a bacterium isolated from spent mushroom substrate. Int J Syst Evol Microbiol 2020; 70:1457–1462 [View Article]
    [Google Scholar]
  13. Makuwa SC, Serepa-Dlamini MH. The antibacterial activity of crude extracts of secondary metabolites from bacterial endophytes associated with Dicoma anomala. Int J Microbiol 2021; 2021:8812043 [View Article] [PubMed]
    [Google Scholar]
  14. MIDI Sherlock Microbial Identification System Operating Manual, Version 6.1 Newark, DE: MIDI Inc; 2008
    [Google Scholar]
  15. Bligh EG, Dyer WJ. A rapid method of total lipid extraction and purification. Can J Biochem Physiol 1959; 37:911–917 [View Article]
    [Google Scholar]
  16. Tindall BJ, Sikorski J, Smibert RM, Kreig NR. Phenotypic characterization and the principles of comparative systematics. Methods Gen Mol Microbiol 2007; 3:330–393 [View Article]
    [Google Scholar]
  17. Tindall BJ, Rosselló-Móra R, Busse HJ, Ludwig W, Kämpfer P. Notes on the characterization of prokaryote strains for taxonomic purposes. Int J Syst Evol Microbiol 2010; 60:249–266 [View Article] [PubMed]
    [Google Scholar]
  18. Vieira S, Huber KJ, Neumann-Schaal M, Geppert A, Luckner M et al. Usitatibacterrugosus gen. nov., sp. nov. and Usitatibacter Palustris sp. nov., novel members of Usitatibacteraceae fam. nov. within the order Nitrosomonadales isolated from soil. Int J Syst Evol Microbiol 2021; 71:004631
    [Google Scholar]
  19. Schumann P, Kalensee F, Cao J, Criscuolo A, Clermont D et al. Reclassification of Haloactinobacterium glacieicola as Occultella glacieicola gen. nov., comb. nov., of Haloactinobacterium album as Ruania alba comb. nov., with an emended description of the genus Ruania, recognition that the genus names Haloactinobacterium and Ruania are heterotypic synonyms and description of Occultellaaeris sp. nov., a halotolerant isolate from surface soil sampled at an ancient copper smelter. Int J Syst Evol Microbiol 2021; 71:004769 [View Article] [PubMed]
    [Google Scholar]
  20. Matsuda N, Matsuda M, Notake S, Yokokawa H, Kawamura Y et al. Evaluation of a simple protein extraction method for species identification of clinically relevant staphylococci by matrix-assisted laser desorption ionization-time of flight mass spectrometry. J Clin Microbiol 2012; 50:3862–3866 [View Article]
    [Google Scholar]
  21. Yoon S-H, Ha S-M, Kwon S, Lim J, Kim Y et al. Introducing EzBioCloud: a taxonomically united database of 16S rRNA gene sequences and whole-genome assemblies. Int J Syst Evol Microbiol 2017; 67:1613–1617 [View Article] [PubMed]
    [Google Scholar]
  22. Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: Molecular Evolutionary Genetics Analysis across computing platforms. Mol Biol Evol 2018; 35:1547–1549 [View Article] [PubMed]
    [Google Scholar]
  23. Felsenstein J. Evolutionary trees from DNA sequences: a maximum likelihood approach. J Mol Evol 1981; 17:368–376 [View Article] [PubMed]
    [Google Scholar]
  24. Saitou N, Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 1987; 4:406–425 [View Article] [PubMed]
    [Google Scholar]
  25. Meier-Kolthoff JP, Göker M. TYGS is an automated high-throughput platform for state-of-the-art genome-based taxonomy. Nat Commun 2019; 10:2182 [View Article] [PubMed]
    [Google Scholar]
  26. Meier-Kolthoff JP, Carbasse JS, Peinado-Olarte RL, Göker M. TYGS and LPSN: a database tandem for fast and reliable genome-based classification and nomenclature of prokaryotes. Nucleic Acids Res 2022; 50:D801–D807 [View Article] [PubMed]
    [Google Scholar]
  27. Lee I, Ouk Kim Y, Park S-C, Chun J. OrthoANI: an improved algorithm and software for calculating average nucleotide identity. Int J Syst Evol Microbiol 2016; 66:1100–1103 [View Article]
    [Google Scholar]
  28. de los Santos Villalobos S, Robles RI, Parra Cota FI, Larsen J, Lozano P et al. Bacillus cabrialesii sp. nov., an endophytic plant growth promoting bacterium isolated from wheat (Triticum turgidum subsp. durum) in the Yaqui Valley, Mexico. Int J Syst Evol Microbiol 2019; 69:3939–3945 [View Article]
    [Google Scholar]
  29. Maughan H, van der Auwera G. Bacillus taxonomy in the genomic era finds phenotypes to be essential though often misleading. Infect Genet Evol 2011; 11:789–797 [View Article] [PubMed]
    [Google Scholar]
  30. Gevers D, Cohan FM, Lawrence JG, Spratt BG, Coenye T et al. Opinion: re-evaluating prokaryotic species. Nat Rev Microbiol 2005; 3:733–739 [View Article] [PubMed]
    [Google Scholar]
  31. Liu Y, Lai Q, Göker M, Meier-Kolthoff JP, Wang M et al. Genomic insights into the taxonomic status of the Bacillus cereus group. Sci Rep 2015; 5:14082 [View Article]
    [Google Scholar]
  32. Soufiane B, Baizet M, Côté J-C. Multilocus sequence analysis of Bacillus thuringiensis serovars navarrensis, bolivia and vazensis and Bacillus weihenstephanensis reveals a common phylogeny. Antonie van Leeuwenhoek 2013; 103:195–205 [View Article]
    [Google Scholar]
  33. Makuwa SC, Serepa-Dlamini MH. Data on draft genome sequence of Bacillus sp. strain MHSD28, a bacterial endophyte isolated from Dicoma anomala. Data Brief 2019; 26:104524 [View Article] [PubMed]
    [Google Scholar]
  34. Anderson I, Sorokin A, Kapatral V, Reznik G, Bhattacharya A et al. Comparative genome analysis of Bacillus cereus group genomes with Bacillus subtilis. FEMS Microbiol Lett 2005; 250:175–184 [View Article] [PubMed]
    [Google Scholar]
  35. Meier-Kolthoff JP, Hahnke RL, Petersen J, Scheuner C, Michael V et al. Complete genome sequence of DSM 30083T, the type strain (U5/41T) of Escherichia coli, and a proposal for delineating subspecies in microbial taxonomy. Stand Genomic Sci 2014; 9:2 [View Article]
    [Google Scholar]
  36. Lee I, Chalita M, Ha S-M, Na S-I, Yoon S-H et al. ContEst16S: an algorithm that identifies contaminated prokaryotic genomes using 16S RNA gene sequences. Int J Syst Evol Microbiol 2017; 67:2053–2057 [View Article] [PubMed]
    [Google Scholar]
  37. Konstantinidis KT, Tiedje JM. Genomic insights that advance the species definition for prokaryotes. Proc Natl Acad Sci U S A 2005; 102:2567–2572 [View Article] [PubMed]
    [Google Scholar]
  38. Goris J, Konstantinidis KT, Klappenbach JA, Coenye T, Vandamme P et al. DNA-DNA hybridization values and their relationship to whole-genome sequence similarities. Int J Syst Evol Microbiol 2007; 57:81–91 [View Article] [PubMed]
    [Google Scholar]
  39. Richter M, Rosselló-Móra R. Shifting the genomic gold standard for the prokaryotic species definition. Proc Natl Acad Sci U S A 2009; 106:19126–19131 [View Article] [PubMed]
    [Google Scholar]
  40. Chun J, Oren A, Ventosa A, Christensen H, Arahal DR et al. Proposed minimal standards for the use of genome data for the taxonomy of prokaryotes. Int J Syst Evol Microbiol 2018; 68:461–466 [View Article] [PubMed]
    [Google Scholar]
  41. Meier-Kolthoff JP, Auch AF, Klenk H-P, Göker M. Genome sequence-based species delimitation with confidence intervals and improved distance functions. BMC Bioinformatics 2013; 14:60 [View Article] [PubMed]
    [Google Scholar]
  42. Kämpfer P. Limits and possibilities of total fatty acid analysis for classification and identification of Bacillus species. Syst Appl Microbiol 1994; 17:86–98 [View Article]
    [Google Scholar]
  43. Song Y, Yang R, Guo Z, Zhang M, Wang X et al. Distinctness of spore and vegetative cellular fatty acid profiles of some aerobic endospore-forming bacilli. J Microbiol Methods 2000; 39:225–241 [View Article] [PubMed]
    [Google Scholar]
  44. Manzulli V, Rondinone V, Buchicchio A, Serrecchia L, Cipolletta D et al. Discrimination of Bacillus cereus group members by MALDI-TOF mass spectrometry. Microorganisms 2021; 9:142 [View Article] [PubMed]
    [Google Scholar]
  45. Kang H, Weerawongwiwat V, Kim JH, Sukhoom A, Kim W. Bacillus songklensis sp. nov., isolated from soil. Int J Syst Evol Microbiol 2013; 63:4189–4195 [View Article] [PubMed]
    [Google Scholar]
  46. Liu Y, Lai Q, Du J, Shao Z. Bacillus zhangzhouensis sp. nov. and Bacillus australimaris sp. nov. Int J Syst Evol Microbiol 2016; 66:1193–1199 [View Article] [PubMed]
    [Google Scholar]
  47. Kang H, Kang J, Cha I, Kim H, Joung Y et al. Bacillus salinus sp. nov., isolated from commercial solar salt. Int J Syst Evol Microbiol 2020; 70:2696–2702 [View Article] [PubMed]
    [Google Scholar]
  48. Song L, Liu H, Wang J, Huang Y, Dai X et al. Bacillus oceani sp. nov., isolated from seawater. Int J Syst Evol Microbiol 2016; 66:796–800 [View Article]
    [Google Scholar]
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