Skip to main content
Log in

Halorubrum amylolyticum sp. nov., a novel halophilic archaeon isolated from a salt mine

  • Original Paper
  • Published:
Antonie van Leeuwenhoek Aims and scope Submit manuscript

Abstract

A pleomorphic and non-motile halophilic archaeon forming light-red pigmented colonies, strain ZC67T, was isolated from the Yuanyongjing Salt Mine, Yunnan, China. Based on similarity search and phylogenetic analysis of the 16S rRNA gene sequence, strain ZC67T belongs to the genus Halorubrum and is closely related to the species of Halorubrum (Hrr.) saccharovorum JCM 8865T, Hrr. persicum C49T, Hrr. halophilum B8T, Hrr. lipolyticum 9-3T, Hrr. salsamenti Y69T and Hrr. depositum Y78T with 16S rRNA gene sequence similarities of 99.0%, 98.7%, 98.5%, 98.4%, 98.1% and 97.7%, respectively. The values of average nucleotide identity (ANI) and average amino-acid identity (AAI) between strain ZC67T and its close relatives were less than 90.5% and 89.3%, respectively. In silico DNA-DNA hybridization (DDH) analysis showed that DNA-DNA relatedness between strain ZC67T and its relatives is less than 45%. Values of ANI, AAI and in silico DDH were clearly below the thresholds used for the delineation of a new species. The major polar lipids of strain ZC67T were similar to other neutrophilic members in the genus Halorubrum containing phosphatidylglycerol, phosphatidylglycerolphosphate methyl ester, phosphatidylglycerol sulfate and sulfated mannosyl-glucosyl-glycerol diether-1. The DNA G+C content was determined to be 66.3 mol% (based on the draft genome). Combined with other diagnostic characteristics, e.g. phenotypic and chemotaxonomic differences, strain ZC67T is concluded to represent a novel species in the genus Halorubrum, for which the name Halorubrum amylolyticum sp. nov. is proposed. The type strain is ZC67T (=CGMCC 1.15718T = JCM 31850T).

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2

Similar content being viewed by others

Abbreviations

MES:

2-Morpholinoethanesulfonic acid

PIPES:

1, 4-Piperazine bis (ethanesulfonic acid)

HEPES:

4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid

CHES:

2-(Cyclohexylamino) ethanesulfonic acid

DMSO:

dimethylsulfoxide

CGMCC:

China General Microbiological Culture Collection Center

JCM:

Japan Collection of Microorganisms

IU:

International Unit of enzyme activity

MEGA:

Molecular Evolutionary Genetics Analysis

References

  • Auch AF, von Jan M, Klenk HP, Göker M (2010) Digital DNA–DNA hybridization for microbial species delineation by means of genome-to-genome sequence comparison. Stand Genomic Sci 2:117–134

    Article  Google Scholar 

  • Bankevich A, Nurk S, Antipov D, Gurevich AA, Kulikov AS, Lesin VM, Nikolenko SI, Pham S, Prjibelski AD, Pyshkin AV, Sirotkin AV, Vyahhi N, Tesler G, Alekseyev MA, Pevzner PA (2012) SPAdes: a new genome assembly algorithm and its applications to single-cell sequencing. J Comput Biol 19:455–477

    Article  CAS  Google Scholar 

  • Boucher Y, Douady CJ, Sharma AK, Kamekura M, Doolittle WF (2004) Intragenomic heterogeneity and intergenomic recombination among haloarchaeal rRNA genes. J Bacteriol 186:3980–3990

    Article  CAS  Google Scholar 

  • Chen S, He J, Zhang J, Xu Y, Huang J, Ke LX (2017a) Halorubrum salsamenti sp. nov., a novel halophilic archaeon isolated from a brine of salt mine. Curr Microbiol 74:1358–1364

    Article  CAS  Google Scholar 

  • Chen S, Xu Y, Ke LX (2017b) Halorubrum trueperi sp. nov., a halophilic archaeon isolated from a salt mine. Int J Syst Evol Microbiol 67:1564–1570

    Article  CAS  Google Scholar 

  • Chen S, Xu Y, Sun S, Chen F, Liu J (2019) Halalkalicoccus subterraneus sp. nov., an extremely halophilic archaeon isolated from a subterranean halite deposit. Antonie Van Leeuwenhoekvan Leeuwenhoek. https://doi.org/10.1007/s10482-019-01241-3

    Article  Google Scholar 

  • Chun J, Oren A, Ventosa A, Christensen H, Arahal DR, da Costa MS, Rooney AP, Yi H, Xu XW, De Meyer S, Trujillo ME (2018) Proposed minimal standards for the use of genome data for the taxonomy of prokaryotes. Int J Syst Evol Microbiol 68:461–466

    Article  CAS  Google Scholar 

  • Cui HL, Tohty D, Zhou PJ, Liu SJ (2006) Halorubrum lipolyticum sp. nov. and Halorubrum aidingense sp. nov., isolated from two salt lakes in Xin-Jiang, China. Int J Syst Evol Microbiol 56:1631–1634

    Article  CAS  Google Scholar 

  • Cui HL, Lin ZY, Dong Y, Zhou PJ, Liu SJ (2007) Halorubrum litoreum sp. nov., an extremely halophilic archaeon from a solar saltern. Int J Syst Evol Microbiol 57:2204–2206

    Article  CAS  Google Scholar 

  • Cui HL, Gao X, Yang X, Xu XW (2010) Halorussus rarus gen. nov., sp. nov., a new member of the family Halobacteriaceae isolated from a marine solar saltern. Extremophiles 14:493–499

    Article  CAS  Google Scholar 

  • de la Haba RR, Corral P, Sánchez-Porro C, Infante-Domínguez C, Makkay AM, Amoozegar MA, Ventosa A, Papke RT (2018) Genotypic and lipid analyses of strains from the archaeal genus Halorubrum reveal insights into their taxonomy, divergence, and population structure. Front Microbiol 9:512

    Article  Google Scholar 

  • De Ley J, Cattoir H, Reynaerts A (1970) The quantitative measurement of DNA hybridization from renaturation rates. Eur J Biochem 12:133–142

    Article  Google Scholar 

  • Dussault HP (1955) An improved technique for staining red halophilic bacteria. J Bacteriol 70:484–485

    CAS  PubMed  PubMed Central  Google Scholar 

  • Felsenstein J (1981) Evolutionary trees from DNA sequences: a maximum likelihood approach. J Mol Evol 17:368–376

    Article  CAS  Google Scholar 

  • Feng J, Zhou PJ, Liu SJ (2004) Halorubrum xinjiangense sp. nov., a novel halophile isolated from saline lakes in China. Int J Syst Evol Microbiol 54:1789–1791

    Article  CAS  Google Scholar 

  • Fullmer MS, Soucy SM, Swithers KS, Makkay AM, Wheeler R, Ventosa A, Goqarten JP, Papke RT (2014) Population and genomic analysis of the Halorubrum. Front Microbiol 5:140

    Article  Google Scholar 

  • Gerhardt P, Murray RGE, Wood WA, Krieg NR (1994) Methods for general and molecular bacteriology. American Society for Microbiology, Washington, D.C.

    Google Scholar 

  • Gupta RS, Naushad S, Fabros R, Adeolu M (2016) A phylogenomic reappraisal of family-level divisions within the class Halobacteria: proposal to divide the order Halobacteriales into the families Halobacteriaceae, Haloarculaceae fam. nov., and Halococcaceae fam. nov., and the order Haloferacales into the families, Haloferacaceae and Halorubraceae fam nov. Antonie Van Leeuwenhoek 109:1521–1523

    Article  Google Scholar 

  • Gutiérrez C, González C (1972) Method for simultaneous detection of proteinase and esterase activities in extremely halophilic bacteria. Appl Microbiol 24:516–517

    PubMed  PubMed Central  Google Scholar 

  • Gutiérrez MC, Castillo AM, Pagaling E, Heaphy S, Kamekura M, Xue Y, Ma Y, Cowan DA, Jones BE, Grant WD, Ventosa A (2008) Halorubrum kocurii sp. nov., an archaeon isolated from a saline lake. Int J Syst Evol Microbiol 58:2031–2035

    Article  Google Scholar 

  • Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl Acids Symp Ser 41:95–98

    CAS  Google Scholar 

  • Kharroub K, Quesada T, Ferrer R, Fuentes S, Aguilera M, Boulahrouf A, Ramos-Cormenzana A, Monteoliva-Sánchez M (2006) Halorubrum ezzemoulense sp. nov., a halophilic archaeon isolated from Ezzemoul sabkha, Algeria. Int J Syst Evol Microbiol 56:1583–1588

    Article  CAS  Google Scholar 

  • Lowe TM, Eddy SR (1997) tRNAscan-SE: a program for improved detection of transfer RNAgenes in genomic sequence. Nucleic Acids Res 25:955–964

    Article  CAS  Google Scholar 

  • Luo R, Liu B, Xie Y, Li Z, Huang W, Yuan J, He G, Chen Y, Pan Q, Liu Y, Tang J, Wu G, Zhang H, Shi Y, Liu Y, Yu C, Wang B, Lu Y, Han C, Cheung DW, Yiu SM, Peng S, Xiaoqian Z, Liu G, Liao X, Li Y, Yang H, Wang J, Lam TW, Wang J (2012) SOAPdenovo2: an empirically improved memory-efficient short-read de novo assembler. Gigascience 1:18

    Article  Google Scholar 

  • Marmur J (1961) A procedure for the isolation of deoxyribonucleic acid from micro-organisms. J Mol Biol 3:208–218

    Article  CAS  Google Scholar 

  • McGenity TJ, Grant WD (1995) Transfer of Halobacterium saccharovorum, Halobacterium sodomense, Halobacterium trapanicum NRC 34041 and Halobacterium lacusprofundi to the genus Halorubrum gen. nov., as Halorubrum saccharovorum comb. nov., Halorubrum sodomense comb. nov., Halorubrum trapanicum comb. nov., and Halorubrum lacusprofundi comb. nov. Syst Appl Microbiol 18:237–243

    Article  Google Scholar 

  • McGenity TJ, Grant WD (2015) Halorubrum. In: Whitman WB (ed) Bergey’s manual of systematics of Archaea and bacteria. Association with Bergey’s Manual Trust, Wiley, pp 1–11

  • McGenity TJ, Oren A (2012) Life in saline environments. In: Bell E (ed) Life at extremes: environments, organisms, and strategies for survival. CABI International, Wallingford, pp 402–437

  • Medlar AJ, Toronen P, Holm L (2018) AAI-profiler: fast proteome-wide exploratory analysis reveals taxonomic identity, misclassification and contamination. Nucleic Acids Res 46:W479–W485

    Article  CAS  Google Scholar 

  • Meier-Kolthoff JP, Göker M, Spröer C, Klenk HP (2013) When should a DDH experiment be mandatory in microbial taxonomy? Arch Microbiol 195:413–418

    Article  CAS  Google Scholar 

  • Minegishi H, Kamekura M, Itoh T, Echigo A, Usami R, Hashimoto T (2010) Further refinement of the phylogeny of the Halobacteriaceae based on the full-length RNA polymerase subunit B’ (rpoB’) gene. Int J Syst Evol Microbiol 60:2398–2408

    Article  Google Scholar 

  • Nawrocki EP, Eddy SR (2013) Infernal 1.1: 100-fold faster RNA homology searches. Bioinformatics 29:2933–2935

    Article  CAS  Google Scholar 

  • Nawrocki EP, Burge SW, Bateman A, Daub J, Eberhardt RY, Eddy SR, Flodden EW, Gardner PP, Jones TA, Tate J, Finn RD (2014) Rfam 12.0: updates to the RNA families database. Nucleic Acids Res 43:D130–137

    Article  Google Scholar 

  • Ochsenreiter T, Pfeifer F, Schleper C (2002) Diversity of Archaea in hypersaline environments characterized by molecular-phylogenetic and cultivation studies. Extremophiles 6:267–274

    Article  CAS  Google Scholar 

  • Oren A, Ventosa A, Grant WD (1997) Proposed minimal standards for description of new taxa in the order Halobacteriales. Int J Syst Bacteriol 47:233–238

    Article  Google Scholar 

  • Oren A, Arahal DR, Ventosa A (2009) Emended descriptions of genera of the family Halobacteriaceae. Int J Syst Evol Microbiol 59:637–642

    Article  Google Scholar 

  • Parte AC (2019) List of prokaryotic names with standing in nomenclature. https://www.bacterio.net

  • Ram Mohan N, Fullmer MS, Makky AM, Wheeler R, Ventosa A, Naor A, Goqarten JP, Papke RT (2014) Evidence from phylogenetic and genome fingerprinting analyses suggests rapidly changing variation in Halorubrum and Haloarcula populations. Front Microbiol 5:143

    Article  Google Scholar 

  • Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425

    CAS  Google Scholar 

  • Schbert M, Lindgreen S, Orlando L (2016) AdapterRemoval v2: rapid adapter trimming, identification, and read merging. BMC Res Notes 9:88

    Article  Google Scholar 

  • Stackebrandt E, Goebel BM (1994) Taxonomic note: a place for DNA–DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology. Int J Syst Bacteriol 44:846–849

    Article  CAS  Google Scholar 

  • 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. Mol Biol Evol 28:2731–2739

    Article  CAS  Google Scholar 

  • Yim KJ, Cha IT, Lee HW, Song HS, Kim KN, Lee SJ, Nam YD, Hyun DW, Bae JW, Rhee SK, Seo MJ, Choi JS, Choi HJ, Roh SW, Kim D (2014) Halorubrum halophilum sp. nov., an extremely halophilic archaeon isolated from a salt-fermented seafood. Antonie Van Leeuwenhoek 105:603–612

    Article  CAS  Google Scholar 

  • Yoon SH, Ha SM, Lim J, Kwon S, Chun J (2017) A large-scale evaluation of algorithms to calculate average nucleotide identity. Antonie Van Leeuwenhoek 110:1281–1286

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Professor Zhu L. Yang from the Kunming Institute of Botany, Chinese Academy of Sciences for the help in sample collection.

Funding

This work was supported by grants from the National Natural Science Foundation of China (31460003), the Anhui Provincial Key Lab. of the Conservation and Exploitation of Biological Resources (591601), the China Scholarship Council (201808340054) and the Department of Education Anhui Province, China.

Author information

Authors and Affiliations

Authors

Contributions

SC conceived the project. SS, FC, YX, JL and SC performed the experiments. SS and SC analysed the data, and drafted the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Shaoxing Chen.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

No specific ethical or institutional permits were required to conduct sampling and the experimental studies did not involve endangered or protected species.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA and rpoB′ genes of strain ZC67T are KX376720 and KX595316, respectively. The accession numbers for the genome sequences of strains ZC67T, Hrr. salsamenti Y69T and Hrr. depositum Y78T are SDJP00000000, VCNL00000000 and VCNM00000000, respectively.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 2595 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sun, S., Chen, F., Xu, Y. et al. Halorubrum amylolyticum sp. nov., a novel halophilic archaeon isolated from a salt mine. Antonie van Leeuwenhoek 112, 1849–1861 (2019). https://doi.org/10.1007/s10482-019-01313-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10482-019-01313-4

Keywords

Navigation