Halobellus captivus sp. nov., an extremely halophilic archaeon isolated from a subterranean salt mine

Abstract

An extremely halophilic archaeon, strain ZY21T, was isolated from a subterranean rock salt sample in Yunnan, China. Colonies of strain ZY21T on nutrient-rich agar plates are orange, wet and transparent. Cells are pleomorphic, motile, Gram-stain negative and lyse in distilled water. Cells can grow at 20–55 °C (optimum 42 °C), in the presence of 15–30% (w/v) NaCl (optimum 18–20%) and at pH 6.0–9.5 (optimum 7.5). Mg2+ is required for growth (optimum 0.3 M). The major polar lipids of strain ZY21T are phosphatidylglycerol, phosphatidylglycerol sulfate and phosphatidylglycerol phosphate methyl ester, sulfated mannosyl-glucosyl-glycerol diether-1 and seven unidentified glycolipids. Sequence similarity searches with the 16S rRNA gene and rpoB′ gene showed that strain ZY21T is closely related to Halobellus rufus CBA1103T (sequence similarities: 97.5% for 16S rRNA gene and 93.3% for rpoB′ gene). The DNA G+C content of strain ZY21T was determined to be 63.0 mol% based on the draft genome sequence. Genome-based sequence similarity analysis showed that the values of the ANI, AAI, and DDH were far below the boundary for delineation of new species. Phenotypic, chemotaxonomic characteristics and phylogenetic properties suggest that strain ZY21T represents a novel species in the genus Halobellus, for which the name Halobellus captivus sp. nov. is proposed. The type strain is ZY21T (= CGMCC 1.16343T = NBRC 113439T).

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Abbreviations

ANI:

Average nucleotide identity

AAI:

Average amino-acid identity

DDH:

DNA–DNA hybridization

CGMCC:

China General Microbiological Culture Collection Center

NBRC:

NTE Biological Resource Center (Japanese)

MES:

2-Morpholinoethanesulfonic acid

PIPES:

1,4-Piperazine bis (ethanesulfonic acid)

CHES:

2-(Cyclohexylamino) ethanesulfonic acid

CAPS:

N-Cyclohexyl-3-aminopropanesulfonic acid

DMSO:

Dimethylsulfoxide

OD:

Optical density

DDH:

DNA–DNA hybridization

TLC:

Thin-layer chromatography

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Acknowledgements

We thank Professor Zhu L. Yang from the Kunming Institute of Botany, Chinese Academy of Sciences for the help in sample collection, and we also thank Dr. Hao Yan from the College of Life Sciences, Anhui Normal University, for his technical assistance.

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.

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

Corresponding author

Correspondence to Shaoxing Chen.

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The authors declare that they have no conflict of interest.

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

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Chen, S., Sun, S., Xu, Y. et al. Halobellus captivus sp. nov., an extremely halophilic archaeon isolated from a subterranean salt mine. Antonie van Leeuwenhoek 113, 221–231 (2020). https://doi.org/10.1007/s10482-019-01332-1

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Keywords

  • Haloarchaea
  • Halobellus
  • Salt mine
  • Salt deposit
  • Polyphasic taxonomy