PMID- 28389550
OWN - NLM
STAT- MEDLINE
DCOM- 20171116
LR  - 20191210
IS  - 1098-5336 (Electronic)
IS  - 0099-2240 (Print)
IS  - 0099-2240 (Linking)
VI  - 83
IP  - 12
DP  - 2017 Jun 15
TI  - Single-Cell Resolution of Uncultured Magnetotactic Bacteria via 
      Fluorescence-Coupled Electron Microscopy.
LID - 10.1128/AEM.00409-17 [doi]
LID - e00409-17
AB  - Magnetotactic bacteria (MTB) form intracellular chain-assembled nanocrystals of 
      magnetite or greigite termed magnetosomes. The characterization of magnetosome 
      crystals requires electron microscopy due to their nanoscopic sizes. However, 
      electron microscopy does not provide phylogenetic information for MTB. We have 
      developed a strategy for the simultaneous and rapid phylogenetic and 
      biomineralogical characterization of uncultured MTB at the single-cell level. It 
      consists of four steps: (i) enrichment of MTB cells from an environmental sample, 
      (ii) 16S rRNA gene sequencing of MTB, and (iii) fluorescence in situ 
      hybridization analyses coordinated with (iv) transmission or scanning electron 
      microscopy of the probe-hybridized cells. The application of this strategy 
      identified a magnetotactic Gammaproteobacteria strain, SHHR-1, from brackish 
      sediments collected from the Shihe River estuary in Qinhuangdao City, China. 
      SHHR-1 magnetosomes are elongated prismatic magnetites which can be idealized as 
      hexagonal prisms. Taxonomic groups of uncultured MTB were also identified in 
      freshwater sediments from Lake Miyun in northern Beijing via this novel 
      coordinated fluorescence and scanning electron microscopy method based on four 
      group-specific rRNA-targeted probes. Our analyses revealed that major 
      magnetotactic taxonomic groups can be accurately determined only with coordinated 
      scanning electron microscopy observations on fluorescently labeled single cells 
      due to limited group coverage and specificity for existing group-specific MTB 
      fluorescence in situ hybridization (FISH) probes. Our reported strategy is simple 
      and efficient, offers great promise toward investigating the diversity and 
      biomineralization of MTB, and may also be applied to other functional groups of 
      microorganisms.IMPORTANCE Magnetotactic bacteria (MTB) are phylogenetically 
      diverse and biomineralize morphologically diverse magnetic nanocrystals of 
      magnetite or greigite in intracellular structures termed magnetosomes. However, 
      many uncultured MTB strains have not been phylogenetically identified or 
      structurally investigated at the single-cell level, which limits our 
      comprehensive understanding of the diversity of MTB and their role in 
      biomineralization. We developed a fluorescence-coupled electron microscopy method 
      for the rapid phylogenetic and biomineralogical characterization of uncultured 
      MTB at the single-cell level. Using this novel method, we successfully identified 
      taxonomic groups of several uncultured MTB and one novel magnetotactic 
      Gammaproteobacteria strain, SHHR-1, from natural environments. Our analyses 
      further indicate that strain SHHR-1 forms elongated prismatic magnetites. Our 
      findings provide a promising strategy for the rapid characterization of 
      phylogenetic and biomineralogical properties of uncultured MTB at the single-cell 
      level. Furthermore, due to its simplicity and generalized methodology, this 
      strategy can also be useful in the study of the diversity and biomineralization 
      properties of microbial taxa involved in other mineralization processes.
CI  - Copyright (c) 2017 Li et al.
FAU - Li, Jinhua
AU  - Li J
AD  - Key Laboratory of Earth and Planetary Physics, Institute of Geology and 
      Geophysics, Chinese Academy of Sciences, Beijing, People's Republic of China 
      lijinhua@mail.iggcas.ac.cn.
AD  - Laboratory for Marine Geology, Qingdao National Laboratory for Marine Science and 
      Technology, Qingdao, People's Republic of China.
AD  - France-China Biomineralization and Nano-Structures Laboratory, Chinese Academy of 
      Sciences, Beijing, People's Republic of China.
FAU - Zhang, Heng
AU  - Zhang H
AD  - Key Laboratory of Earth and Planetary Physics, Institute of Geology and 
      Geophysics, Chinese Academy of Sciences, Beijing, People's Republic of China.
AD  - Laboratory for Marine Geology, Qingdao National Laboratory for Marine Science and 
      Technology, Qingdao, People's Republic of China.
AD  - France-China Biomineralization and Nano-Structures Laboratory, Chinese Academy of 
      Sciences, Beijing, People's Republic of China.
AD  - College of Life Science and Technology, Heilongjiang Bayi Agricultural 
      University, Daqing, People's Republic of China.
FAU - Menguy, Nicolas
AU  - Menguy N
AD  - France-China Biomineralization and Nano-Structures Laboratory, Chinese Academy of 
      Sciences, Beijing, People's Republic of China.
AD  - IMPMC, CNRS UMR 7590, Sorbonne Universites, MNHN, UPMC, IRD UMR 206, Paris, 
      France.
FAU - Benzerara, Karim
AU  - Benzerara K
AD  - IMPMC, CNRS UMR 7590, Sorbonne Universites, MNHN, UPMC, IRD UMR 206, Paris, 
      France.
FAU - Wang, Fuxian
AU  - Wang F
AD  - Key Laboratory of Earth and Planetary Physics, Institute of Geology and 
      Geophysics, Chinese Academy of Sciences, Beijing, People's Republic of China.
AD  - Laboratory for Marine Geology, Qingdao National Laboratory for Marine Science and 
      Technology, Qingdao, People's Republic of China.
AD  - France-China Biomineralization and Nano-Structures Laboratory, Chinese Academy of 
      Sciences, Beijing, People's Republic of China.
FAU - Lin, Xiaoting
AU  - Lin X
AD  - Key Laboratory of Earth and Planetary Physics, Institute of Geology and 
      Geophysics, Chinese Academy of Sciences, Beijing, People's Republic of China.
AD  - College of Life Science and Technology, Heilongjiang Bayi Agricultural 
      University, Daqing, People's Republic of China.
FAU - Chen, Zhibao
AU  - Chen Z
AD  - College of Life Science and Technology, Heilongjiang Bayi Agricultural 
      University, Daqing, People's Republic of China.
FAU - Pan, Yongxin
AU  - Pan Y
AD  - Key Laboratory of Earth and Planetary Physics, Institute of Geology and 
      Geophysics, Chinese Academy of Sciences, Beijing, People's Republic of China.
AD  - France-China Biomineralization and Nano-Structures Laboratory, Chinese Academy of 
      Sciences, Beijing, People's Republic of China.
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
DEP - 20170531
PL  - United States
TA  - Appl Environ Microbiol
JT  - Applied and environmental microbiology
JID - 7605801
SB  - IM
MH  - Bacterial Typing Techniques/instrumentation/*methods
MH  - China
MH  - Gammaproteobacteria/classification/genetics/*isolation & 
      purification/ultrastructure
MH  - Genome, Bacterial
MH  - Geologic Sediments/*microbiology
MH  - In Situ Hybridization, Fluorescence
MH  - Lakes/*microbiology
MH  - Magnetosomes/chemistry/genetics/ultrastructure
MH  - Microscopy, Electron
MH  - Phylogeny
MH  - Rivers/*microbiology
MH  - Single-Cell Analysis/instrumentation/*methods
PMC - PMC5452806
OTO - NOTNLM
OT  - FISH
OT  - TEM
OT  - biomineralization
OT  - fluorescence-coupled electron microscopy
OT  - magnetotactic bacteria
EDAT- 2017/04/09 06:00
MHDA- 2017/11/29 06:00
PMCR- 2017/05/31
CRDT- 2017/04/09 06:00
PHST- 2017/02/17 00:00 [received]
PHST- 2017/04/05 00:00 [accepted]
PHST- 2017/04/09 06:00 [pubmed]
PHST- 2017/11/29 06:00 [medline]
PHST- 2017/04/09 06:00 [entrez]
PHST- 2017/05/31 00:00 [pmc-release]
AID - AEM.00409-17 [pii]
AID - 00409-17 [pii]
AID - 10.1128/AEM.00409-17 [doi]
PST - epublish
SO  - Appl Environ Microbiol. 2017 May 31;83(12):e00409-17. doi: 10.1128/AEM.00409-17. 
      Print 2017 Jun 15.