Fig. S3. Polar Lipids of Strain T16B2T and All Nine Type Strains of the Genus Hyphomonas

2 Fig. S1. Transmission electron micrographs of negatively stained strains T16B2T cells grown

3 on marine agar medium for 72 h.

10 11

12 Fig. S2. Rep-PCR analyis of strains T16B2T, T24B3 and C76AD. (lane 1, T16B2T; lane 2,

13 T24B3; Lane 3, C76AD. M, DNA molecular weight marker: from top, 10kb, 9kb, 8 kb, 7 kb, 6

14 kb, 5 kb, 4 kb, 3 kb, 2 kb, 1 kb,).

23 Fig. S3. Polar lipids of strain T16B2T and all nine type strains of the genus Hyphomonas 24

25 1. 2.

27 3. 4. 28

29 5. 6.

31 7. 8. 32

33 9. 10.

34 Strains: 1, T16B2T; 2, H. beringensis 25B14_1T; 3, H. chukchiensis BH-BN04-4T; 4, H.

35 atlanticus 22II1-22F38T; 5, H. polymorpha DSM 2665T; 6, H. neptunium DSM 5154T; 7, H.

36 jannaschiana DSM 5153T; 8, H. oceanitis DSM 5155T; 9, H. johnsonii ATCC 43964T; 10, H.

37 adhaerens ATCC 43965T. GL, unidentified glycolipid; PL, unidentified phospholipid; P,

38 unknown polar lipid.

40 41 Table S1. Differential susceptibility to antibiotics of strains T16B2T, T24B3 and C76AD and

42 related Hyphomonas species.

43 Strains: 1, T16B2T; 2, T24B3; 3, C76AD; 4, H. beringensis 25B14_1T; 5, H. chukchiensis BH-

44 BN04-4T; 6, H. atlanticus 22II1-22F38T; 7, H. polymorpha DSM 2665T; 8, H. neptunium DSM

45 5154T; 9, H. jannaschiana DSM 5153T; 10, H. oceanitis DSM 5155T; 11, H. johnsonii ATCC

46 43964T; 12, H. adhaerens ATCC 43965T. The data for strains T16B2T, T24B3 and C76AD were

47 determined from this study. Other data were retrieved from our previous work (Li et al. 2014a,

48 Li et al. 2014c). And the strains used in this study were test at the same conditions with our

49 previous study (Li et al. 2014a, Li et al. 2014c). Characteristics are scored as: S, sensitive; R,

50 resistant.

Susceptibility to 1 2 3 4 5 6 7 8 9 10 11 12

antibiotics Ampicillin (10 μg) S S S S R S S R S R S R Cephradin (30 μg) S S S S S S S S R R S R Cefobid (30 μg) R R R R R R S R R R R R Ciprofloxacin (5 μg) S S S S S R S R S R S S Cefalexin (30 μg) S S S S S S S S R R S R Gentamicin (10 μg) S S S S S S R R S S S S Rocephin (30 μg) S S S S S S R S S R S S Erythromycin (15 μg) S S S S S S R S S S S S Cefazolin (30 μg) S S S S R S S R R R S R Norfloxacin (10 μg), S R S R R R R R R R R R Penicillin G (10 μg) S S R R R S R R S R S R Piperacillin (100 μg) S S S S S S S S S R S S Streptomycin (10 μg) S R S S R R R R S S S S Furazolidone (15 μg) S S S S S S S S S R R R Neomycin (10 μg) S S S R S R R S S S S S 51 52 Table S2. Summary of genomic features of strain T16B2T and nine type strains of the genus Hyphomonas

GenBank accession Size Gene Pnumber of rRNA number Strains numbers* (Mb) s of tRNA T16B2T AWFA00000000 3.42 3345 33 41 H. beringensis 25B14_1T AWFF00000000 3.80 3660 33 41 H. chukchiensis BH-BN04-4T AWFG00000000 3.86 3823 33 42 H. atlanticus 22II1-22F38T AWFH00000000 3.60 3561 33 40 H. polymorpha DSM 2665T ARYM00000000 4.05 3893 31 41 H. neptunium DSM 5154T CP000158.1 3.71 3568 33 43 H. jannaschiana DSM 5153T ARYJ00000000 3.64 3546 31 40 H. oceanitis DSM 5155T ARYL00000000 4.28 4216 41 43 H. johnsonii ATCC 43964T ARYK00000000 3.62 3489 31 41 H. adhaerens ATCC 43965T ARYH00000000 3.67 3522 31 42 53 *The GenBank accession numbers were obtained from reference (Li et al. 2014b). 54 Table S3. Fatty acids of strains T16B2T, T24B3 and C76AD and nine type strains of the genus

55 Hyphomonas.

56 Strains: 1, T16B2T; 2, T24B3; 3, C76AD; 4, H. beringensis 25B14_1T; 5, H. chukchiensis BH-

57 BN04-4T; 6, H. atlanticus 22II1-22F38T; 7, H. polymorpha DSM 2665T; 8, H. neptunium DSM

58 5154T; 9, H. jannaschiana DSM 5153T; 10, H. oceanitis DSM 5155T; 11, H. johnsonii ATCC

59 43964T; 12, H. adhaerens ATCC 43965T. The data for strains T16B2T, T24B3 and C76AD were

60 determined in this study. Other data were retrieved from our previous reports ( Li et al . 2014a,

61 Li et al. 2014c). All strains were tested under the same conditions. Values are percentages of

62 total fatty acids. ND, not detected; tr, trace amount (<1.0%). Fatty acids less than 1.0% of all

63 strains are not shown.

Fatty acids 1 2 3 4 5 6 7 8 9 10 11 12

C9:0 ND ND ND ND ND ND ND ND 4.6 ND ND ND

C12:1 3OH ND 2.0 2.2 2.0 2.2 ND 1.7 1.7 2.4 2.4 1.8 1.7

C16:0 9.7 14.1 13.5 16.0 14.5 8.9 2.9 8.3 10.4 20.2 18.4 16.2

C16:1ω5c 1.6 1.2 tr 2.7 4.5 2.0 tr 2.0 2.4 7.2 5.0 1.2

C17:0 7.1 7.4 5.4 7.4 1.3 5.0 15.7 3.8 6.6 tr 2.8 3.2

C17:1ω6c 4.5 3.7 2.0 4.3 2.2 7.5 23.5 8.0 7.0 tr 3.4 2.2

C17:1ω8c 3.0 3.7 3.0 3.1 2.5 5.9 21.5 15.8 6.1 tr 3.6 3.0

C18:1ω5c tr tr tr 1.1 tr tr tr tr tr 1.0 tr tr

C18:1ω7c-methyl 14.7 10.2 12.2 16.8 7.7 8.6 1.9 4.0 11.8 16.2 4.8 11.6

C18:0 4.7 7.0 4.8 4.1 tr 2.3 tr 2.2 2.5 1.7 4.7 6.2

C18:1ω9c tr tr tr tr ND 1.2 ND 1.1 tr tr tr tr

C19:0ω8c-cyclo ND ND ND ND ND 1.4 ND tr ND 1.0 ND tr

C20:0 1.0 1.2 1.3 tr tr 1.2 ND tr tr 1.0 tr tr Summed tr tr tr tr 3.6 1.3 tr 3.5 tr 4.8 2.8 tr Feature 3 Summed tr tr tr tr tr tr 2.4 1.0 tr tr tr tr Feature 7 Summed 49.7 43.9 51.6 38.3 58.0 49.9 25.4 45.1 42.1 40.3 47.4 49.1 Feature 8 64 *Summed features represent groups of two or three fatty acids that could not be separated by 65 GLC with the MIDI system. Summed Feature 3, C16:1ω6c/ω7c; Summed Feature 7,

66 unidentified 18.846/C19:1ω6c; summed Feature 8, C18:1ω7c/ω6c.