Supplementary Material

Table S1 Housekeeping genes primers, PCR conditions.

primer / Sequences 5´-3ˊ / PCR condition
atpD-f / CCAACCATGTAGAASGCYTG / 94°C, 5 min, (94°C, 40S; 55°C 40S; 72°C, 90S)× 30 cycles, 72°C,10 min.
atpD-r / GAAGTTCAGCARCAGCTG
gyrB-f / GGYGGTAAGTTCGAYGAYAAYWSCTAYAARG / 94°C, 5 min, (94°C, 40S; 58°C 40S; 72°C, 90S)× 30 cycles, 72°C,10 min.
gyrB-r / CCRTCVACRTCRGCATCGGTCATG
rpoB-f※ / TRRTCVGCTTCTTCRATRG / 94°C, 5 min, (94°C, 40S; 48°C 40S; 72°C, 60S)× 30 cycles, 72°C,5 min.
rpoB-r※ / CCGTCGTATYCGYTSTGT

※ Primers rpoB were not suitable for Oleibacter marinus NBRC105760T.

Table S2 Fatty acid compositions (%) of strain GYP-2T and the type strains of close related taxa in the family Oceanospirillaceae..

Strains: 1, GYP-2T; 2, Oceanobacter kriegii LMG 6238T; 3, Oleibacter marinus NBRC 105760T; 4, Thalassolituus oleivorans LMG 21420T. –, < 1% of total fatty acid content.

Fatty acid / 1 / 2 / 3 / 4
C10:0 / – / – / 9.52 / –
C11:0 / – / – / 1.91 / –
C12:0 / 6.35 / 9.20 / 23.84 / 1.13
C14:0 / – / – / 2.98 / 3.26
C16:0 / 18.81 / 19.71 / 13.54 / 20.06
C18:0 / 1.67 / 2.79 / 2.02 / –
C10:03-OH / 2.99 / 3.86 / 0.97 / –
C12:03-OH / 2.43 / 4.32 / 12.71 / 4.89
Iso C11:03-OH / – / – / 1.51 / –
Iso C12:0 / – / – / 1.72 / –
Cyclo C17:0 / – / – / – / 5.53
Cyclo C19:0ω8c / – / – / – / 17.95
C17:1ω8c / 1.29 / – / 1.75 / –
C18:1ω9c / – / – / 10.61 / –
Summed feature 2※ / – / – / 3.25 / –
Summed feature 3※ / 25.30 / 26.40 / 10.28 / 3.46
Summed feature 4※ / – / 1.13 / 2.10 / –
Summed feature 8※ / 35.09 / 29.34 / 1.28 / 40.47

※Summed feature 2: C12:0 aldehyde, unknown ECL 10.928, isoC16:1I and/or C14:03-OH; Summed feature 3: C16:1ω7c and/or C16:1ω6c; Summed feature 4: isoC17:1I and/or anteiso B; Summed feature 8: C18:1ω6c and/or C 18:1ω7c.

Fig. S1 1H NMR spectrum of PHB produced by GYP-2T. (Solvent=DCCl3)

Fig. S2 Transmission electron micrograph of negatively stained cells of strain GYP-2T. Cells were grown on GYP plate for 2 days at 28 °C

Fig. S3 Maximum-parsimony phylogenetic tree based on 16S rRNA gene sequences showing the relationship between strain GYP-2T and type species in related genera. Numbers at nodes indicate percentages of 1000 bootstrap resamplings, only values above 50 % are shown.

Fig. S4 Maximum likelihood phylogenetic tree based on 16S rRNA gene sequences showing the relationship between strain GYP-2T and type species in related genera from Gammaproteobacteria. Numbers at nodes indicate percentages of 1000 bootstrap resamplings, only values above 50 % are shown. Bar, 0.01 substitutions per site.

Fig. S5 Maximum likelihood phylogenetic trees based on housekeeping genes atpD, gyrB and rpoB deduced amino acid sequences showing the relationship between strain GYP-2T and type species in related genera. Numbers at nodes indicate percentages of 1000 bootstrap resamplings. Bars, 0.01 or 0.05 substitutions per site.

Fig. S6 HPLC spectrum of the respiratory quinone. GYP-2T retention time =13.53 min, should be Q-9; Oceanobacter kriegii LMG 6238T retention time =10.43 min, should be Q-8.

Fig. S7 Two-dimensional thin-layer chromatogram of polar lipids of strain GYP-2T and Oceanobacter kriegii LMG 6238T. DPG, diphosphatidylglycerol; PG, phosphatidylglycerol; PE, phosphatidylethanolamine; NPPL, ninhydrin-positive phospholipid; NPL, ninhydrin-positive lipid; PL, unidentified phospholipid; L, unidentified polar lipid.