Supplementary Fig.S1.Agarose gel electrophoresis of housekeeping genes (Lane: M, 100 bp marker; 1, atpD gene; 2, recA gene; 3, glnII gene).

Supplementary Fig.S2. Two-dimensional thin-layer chromatographyseparation of the polar lipids of strainNHI-8T. 1,Cardiolipin; 2,phosphatidylglycerol; 3, ornithine-containing lipid; 4, phosphatidylethanolamine; 5,phosphatidyl-N-dimethylethanolamine; and 6,phosphatidylcholine.

Supplementary Fig.S3. SDS-PAGE of whole-cell soluble proteins of strain NHI-8T and type strains of closely related Mesorhizobium species. Lanes: 1, Marker (kDa); 2, M. soliNHI-8T; 3,M. chacoenseKACC13891T; 4,M. mediterraneumKACC10664T; 5,M. robiniaeHAMBI3082T; 6,M. muleienseHAMBI3264T; 7,M. temperatumHAMBI2583T; 8,M. tamadayenseLMG26736T; 9,M. gobienseHAMBI2974T; 10,M. tianshanenseKACC10665T; 11,M. opportunistumHAMBI3007T; 12,M. lotiKACC11199T; 13,M. huakuiiKACC10731T; 14,M. metalliduransLMG24485T; and 15,M. amorphaeKACC11070T.

Supplementary Table S1.Isolates and reference strains used in this study.

Strain / Collection number / Original host plant / Geographic origin / References
M. soliNHI-8T / KEMB9005-153T
KACC17916T
JCM19897T / Robiniapseudoacacia / South Korea / This study
M. chacoense / KACC13891T / Prosopis alba / Argentina / Velázquezet al. 2001
M. mediterraneum / KACC10664T / Cicerarietinum / Syria / Nouret al. 1995; Jarvis et al. 1997
M. robiniae / HAMBI3082T / Robiniapseudoacacia / China / Zhouet al. 2010
M. muleiense / HAMBI3264T / Cicerarietinum / China / Zhanget al. 2012
M. temperatum / HAMBI2583T / Astragalusadsurgens / China / Gaoet al. 2004
M. tamadayense / LMG26736T / Anagyrislatifolia and Lotus berthelotii / Spain / Ramírez-Bahenaet al. 2012
M. gobiense / HAMBI2974T / ? / China / Hanet al. 2008
M. tianshanense / KACC10665T / Glycyrrhizapallidiflora / China / Chenet al. 1995, Jarvis et al. 1997
M. opportunistum / HAMBI3007T / Biserrulapelecinus / Australia / Nandasenaet al. 2009
M. loti / KACC11199T / Lotus spp. / New Zealand / Jarvis et al. 1997, Jarvis et al. 1982
M. huakuii / KACC10731T / Astragalussinicus / China / Chen et al. 1991, Jarvis et al. 1997
M. metallidurans / LMG24485T / Anthyllisvulneraria / France / Vidal et al.2009

KEMB: Korea National Environmental Microorganisms Bank; KACC: Korean Agricultural Culture Collection; LMG: Laboratory of Microbiology, Department of Biochemistry and Microbiology, Faculty of Sciences of Ghent University of the Belgian Coordinated Collections of Microorganisms; HAMBI: University of Helsinki, Faculty of Agriculture and Forestry, Division of Microbiology and Biotechnology.

Supplementary Table S2.Profiles and relative content of fatty acids in strain NHI-8T and type strains of closely related Mesorhizobium species.

Fatty acid (relative content, %) / 1 / 2 / 3 / 4 / 5 / 6 / 7 / 8 / 9 / 10 / 11 / 12 / 13
Straight-chain saturated
C12:0 / 0.4 / 0.4 / 0.2 / 1.8 / 1.9 / 0.8 / 0.12 / 0.3 / - / 2.6 / - / - / 1.6
C14:0 / - / - / - / - / - / - / - / 0.2 / - / - / - / - / -
C16:0 / 5.0 / 7.3 / 10.2 / 14.7 / 7.8 / 10.2 / 13.1 / 11.7 / 9.0 / 13.2 / 14.6 / 13.2 / 12.1
C17:0 / 0.4 / 0.3 / 0.6 / - / 0.3 / 0.5 / 0.3 / - / - / - / - / - / -
C18:0 / 11.0 / 8.3 / 4.6 / 5.9 / 7.1 / 6.3 / 6.0 / 4.5 / 2.9 / 4.9 / 0.5 / 4.4 / 4.9
C19:0 / - / - / 0.1 / - / - / - / - / - / - / - / - / - / -
C20:0 / - / - / 0.2 / 1.4 / - / - / 0.6 / - / - / 0.5 / 0.4 / 0.9 / -
Branched saturated
iso-C10:0 / 0.2 / 0.3 / 0.2 / 0.8 / 0.5 / 0.6 / 0.2 / 0.4 / 1.6 / 0.5 / 0.2 / 0.7
anteiso-C11:0 / - / - / - / - / - / - / - / - / 1.4 / - / - / - / -
C12:03OH / - / - / 0.3 / - / 0.4 / 0.4 / 0.3 / 0.2 / - / 0.6 / 0.3 / 0.3 / 0.6
iso-C13:03OH / - / - / 0.3 / 0.8 / 0.4 / - / 0.6 / 0.5 / - / - / 0.3 / 0.4 / -
anteiso-C13:0 / - / - / - / - / - / - / - / - / 1.4 / - / - / - / -
iso-C15:03OH / 0.3 / 0.9 / - / - / - / - / - / - / - / - / - / - / -
iso-C15:0 / - / 1.6 / 0.2 / - / - / - / - / 0.3 / - / - / 0.3 / 0.2 / -
anteiso-C15:0 / - / - / - / - / 0.5 / - / - / - / 0.9 / - / - / 0.2 / -
alcohol-C16:0 N / 0.3 / 0.4 / 0.2 / 1.7 / 0.6 / 0.8 / 0.2 / 0.4 / 1.2 / 1.4 / - / 0.2 / 1.0
iso-C17:0 / 5.0 / 7.5 / 3.2 / 5.2 / 3.3 / 3.5 / 2.8 / 4.6 / 3.0 / 5.2 / 8.2 / 4.7 / 6.2
cyclo-C17:0 / - / 0.5 / 0.9 / - / - / - / 0.8 / 0.5 / - / 1.3 / 0.6 / 0.6 / -
anteiso-C17:0 / - / - / - / - / - / - / - / - / 0.6 / - / - / 0.2 / -
C18:03OH / 0.3 / - / - / - / - / - / - / - / - / - / - / - / -
iso-C19:0 / 0.5 / - / - / - / - / - / - / - / - / - / - / - / -
cyclo-C19:0ω8c / 17.3 / 11.6 / 34.1 / 17.7 / 13.4 / 16.9 / 11.0 / 10.2 / 5.8 / 36.3 / 20.9 / 30.0 / 8.9
C19:010-methyl / - / - / 0.4 / - / 0.9 / - / 0.5 / 0.4 / - / - / 0.5 / 0.9 / -
Unsaturated
C15:1ω8c / - / - / 0.2 / 0.6 / - / - / 0.4 / 0.4 / - / - / - / 0.2 / -
C17:1ω8c / - / - / - / - / - / - / 0.2 / - / - / - / - / - / -
C18:1ω9c / - / - / 0.6 / 0.8 / - / 1.2 / 0.5 / - / - / - / 5.7 / 2.2 / -
C18:1ω7c 11-methyl / 0.4 / 24.7 / 8.0 / 4.2 / 4.2 / 6.4 / - / 8.7 / 7.2 / 6.7 / 12.6 / 5.3 / 5.9
C18:12OH / - / - / - / - / - / - / - / - / - / - / 0.2 / 0.9 / -
C20:2ω6,9c / 0.5 / 0.3 / 0.5 / 1.2 / 0.5 / 0.7 / 0.3 / - / - / 2.2 / - / 0.5 / -
C20:4ω6,9,12,15c / - / - / - / - / - / - / - / - / 1.8 / - / - / - / -
C20:1ω7c / 0.5 / 0.3 / 1.4 / - / - / 0.5 / 0.4 / 0.5 / - / - / - / 0.5 / -
Unknown / 0.3 / - / - / - / - / - / - / - / - / - / - / - / -
Summed featuresa
3 / 0.4 / 0.7 / 0.8 / - / 0.8 / 0.7 / 1.1 / 1.4 / 1.8 / - / 0.6 / 0.4 / 1.6
8 / 57.0 / 35.1 / 33.0 / 43.3 / 57.6 / 50.7 / 60.6 / 54.7 / 63.0 / 23.2 / 33.7 / 33.5 / 56.6

aSummed features are groups of two fatty acids that cannot be separated by GLC with the MIDI system. Summed features3 and 8 containedC16:1ω7c/ or C16:1ω6c and C18:1ω7c/ or C18:1ω6c.

Species:1,M. soliNHI-8T; 2,M. chacoenseKACC13891T; 3,M. mediterraneumKACC10664T; 4,M. robiniaeHAMBI3082T; 5,M. muleienseHAMBI3264T; 6,M. temperatumHAMBI2583T; 7,M. tamadayenseLMG26736T; 8,M. gobienseHAMBI2974T; 9,M. tianshanenseKACC10665T; 10,M. opportunistumHAMBI3007T; 11,M. lotiKACC11199T; 12,M. huakuiiKACC10731T; and13,M. metalliduransLMG24485T.-, Not detected.

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