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Algimonas porphyrae gen. nov., sp. nov., a member of the family Hyphomonadaceae, isolated from a red alga Porphyra yezoensis.
Article in International Journal of Systematic and Evolutionary Microbiology · March 2012 DOI: 10.1099/ijs.0.040485-0 · Source: PubMed
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1 Algimonas porphyrae gen. nov., sp. nov., a member of the family Hyphomonadaceae,
2 isolated from a red alga Porphyra yezoensis.
3
4 Youhei Fukui1, Mahiko Abe2, Masahiro Kobayashi3, Hiroaki Saito1, Hiroshi Oikawa4,
5 Yutaka Yano 5, and Masataka Satomi1*.
6
7 1 National Research Institute of Fisheries Science, Fisheries Research Agency,
8 Yokohama, 236-8648, Japan
9 2 National Fisheries University, Shimonoseki, 759-6595, Japan
10 3 Seikai National Fisheries Research Institute, Nagasaki, 851-2213, Japan
11 4National Research Institute of Fisheries and Environment of Inland Sea, Fisheries
12 Research Agency, Hiroshima, 739-0452, Japan
13 5National Salmon Resources Center, Fisheries Research Agency, Sapporo, 062-0922,
14 Japan
15 *Author for correspondence: Masataka Satomi. Tel. & Fax: +81-45-788-7669
16 e-mail: [email protected].
17 National Research Institute of Fisheries Science, Fisheries Research Agency, 2-12-4,
18 Fukuura, Kanazawa-ku, Yokohama, Kanagawa 236-8648, Japan
19 Subjective category: Proteobacteria
20 Running title: Algimonas porphyrae gen. nov., sp. nov.
21
22 The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene sequences of
23 strains 0C-2-2T, 0C-17, and LNM-3 are AB689189, AB689190, and AB689191,
24 respectively. 25 Summary
26 Three Gram-negative, stalked, motile bacteria, designated 0C-2-2T, 0C-17, and LNM-3,
27 were isolated from a red alga Porphyra yezoensis. 16S rRNA gene sequence analysis
28 revealed that the three novel strains belonged in the family Hyphomonadaceae, and are
29 closely related to Litorimonas taeanensis G5T (96.5 % 16S rRNA gene sequence
30 similarity) and Hellea balneolensis 26III/A02/215T (94.3 % similarity). The DNA G+C
31 contents of the new isolates (58.5-60.2 mol%) were clearly distinguished from L.
32 taeanensis G5 T (47.1 mol%) and H. balneolensis DSM 19091T (47.9 mol%). The G+C
33 content of L. taeanensis G5 T obtained in this study was distinct from a previous report
34 (63.6 mol%). DNA-DNA hybridization experiments showed that the new strains
35 constituted a single species. Eleven phenotypic features of the three isolates differed
36 from those of both related genera. The predominant respiratory quinone was
37 ubiquinone-10 and the major fatty acid was C18:1ω7c. On the basis of polyphasic
38 taxonomic analysis, the novel strains represent a novel genus and species for which the
39 name Algimonas porphyrae gen. nov., sp. nov. is proposed, with the type strain 0C-2-2T
40 (= LMG 26424T = NBRC 108216T).
41
42
43
44
45
46
47
48 49 The families Caulobacteraceae and Rhodobacteraceae have been classified into the
50 order Caulobacterales and Rhodobacterales within the class Alphaproteobacteria,
51 respectively (Garrity et al., 2005). However, the family Rhodobacteraceae was
52 integrated into the order Caulobacterales on the basis of 16S rRNA gene sequences, and
53 the family Hyphomonadaceae was newly founded in the order Caulobacterales (Lee et
54 al., 2005). The family Hyphomonadaceae included four genera of Hyphomonas,
55 Hirschia, Maricaulis, and Oceanicaulis (Lee et al., 2005) which were previously
56 classified as the family Rhodobacteraceae (Strömpl et al., 2003; Garrity et al., 2005).
57 Subsequently, additional genera, including Hellea, Henriciella, Litorimonas, Ponticaulis,
58 Robiginitomaculum, and Woodsholea have been classified as members of the family
59 Hyphomonadaceae (http://www.bacterio.cict.fr). Most bacteria of the family
60 Hyphomonadaceae are characterized by having a single or two stalks. These type strains
61 have been isolated from marine habitats such as seawater (Lee et al., 2007; Alain et al.,
62 2008), brackish water (Schlesner et al., 1990), hydrothermal vent (Weiner et al., 2000),
63 beach sand (Jung et al., 2011), shellfish beds (Weiner et al., 1985), and dinoflagellates
64 (Strömpl et al., 2003). In this study, three strains designed as 0C-2-2T, 0C-17, and
65 LNM-3, were isolated from a red alga Porphyra yezoensis. We describe the
66 phylogenetic, genetic, phenotypic, and chemotaxonomic characters of the three novel
67 strains.
68
69 Thalli of P. yezoensis were cultured in a sterile modified half-strength SWM-III medium
70 (Ogata, 1970) at 17 °C. The samples of the culture medium and the thalli were
71 incubated on Marine agar 2216 (MA; Difco) at 20 °C for 2 weeks. Strains 0C-2-2T and
72 0C-17 were isolated from each culture medium in separate lots. Strain LNM-3 was 73 isolated from a thallus surface. Litorimonas taeanensis G5T and Hellea balneolensis
74 DSM 19091T were used as reference strains in all following tests. MA and Marine broth
75 2216 (MB; Difco) were used for the routine culture of all strains. Novel strains were
76 maintained at 20 °C. L. taeanensis G5T and H. balneolensis DSM 19091T were
77 maintained at 25 °C. All strains were stored at -80 °C in 20 % (v/v) glycerol.
78
79 DNA was prepared by the method of Johnson (1981). The 16S rRNA genes of the three
80 strains were amplified using universal primers of 27F and 1492R (Weisburg et al.,
81 1991). The almost-complete 16S rRNA gene sequences of strains 0C-2-2T (1331 bp),
82 0C-17 (1349 bp), and LNM-3 (1331 bp) were determined using six sequencing primers
83 27F, 519F, 1190F, 530R, 760R, and 1492R (Satomi et al., 1997) using an automated
84 DNA sequencer (model 3100; Applied Biosystems). The sequences close to the novel
85 strains were obtained from the BLAST-N algorithm (Altschul et al., 1990) in the
86 GenBank, EMBL, and DDBJ databases. The 16S rRNA gene sequences of novel strains
87 and the related strains were aligned using Clustal X programs (Thompson et al., 1997)
88 and the sequence similarities were calculated except for alignment gaps and ambiguous
89 bases, using MEGA version 4 (Tamura et al., 2007). Phylogenetic trees were
90 constructed according to three methods of neighbour-joining, maximum-likelihood, and
91 maximum-parsimony analyses. Neighbour-joining tree was constructed with a bootstrap
92 through 1000 replications based on Kimura’s 2 parameter model (Kimura, 1980) using
93 MEGA version 4 (Tamura et al., 2007). Maximum-likelihood and maximum-parsimony
94 trees were constructed using PHYLIP version 3.69 (Felsenstein, 2009). The similarities
95 of 16S rRNA gene sequences among the three strains were 100 %. The 16S rRNA gene
96 sequence of strain 0C-2-2T had the similarities of 96.5 % and 94.3 % with L. taeanensis 97 G5T and H. balneolensis 26III/A02/215T, respectively. The phylogenetic tree based on
98 neighbour-joining analysis showed that the three strains belonged in the family
99 Hyphomonadaceae, and formed an independent group with a high bootstrap value of
100 100 % (Fig. 1). The clustering formation was also recovered in the maximum-likelihood
101 and the maximum-parsimony analyses.
102
103 DNA G+C content was determined using a HPLC with a reversed-phase column
104 (COSMOSIL 5C18-MS-II, Nacalai Tesque) according to the method of Tamaoka &
105 Komagata (1984). DNA of Tetragenococcus halophilus industrial strain (Bio’c) was
106 used as a control in the experiment. DNA-DNA hybridization was performed by a
107 microplate hybridization method (Ezaki et al., 1989). Each DNA of strain 0C-2-2T, L.
108 taeanensis G5T, and H. balneolensis DSM 19091T was labeled with a photobiotin. The
109 DNA G+C contents and the DNA-DNA relatedness among the three strains, L.
110 taeanensis G5T, and H. balneolensis DSM 19091T are shown in Table 1. The G+C
111 contents of the three isolates ranged from 58.5 to 60.2 mol%. The values of L.
112 taeanensis G5T and H. balneolensis DSM 19091T were 47.1 and 47.9 mol%,
113 respectively. The value of H. balneolensis DSM 19091Twas similar to 46.8 mol%
114 (original value, Alain et al., 2008), but the value of L. taeanensis G5T measured in this
115 study was distinct from 63.6 mol% (original value, Jung et al., 2011). The content of T.
116 halophilus industrial strain was 36.1 mol%, which was similar to a previous value by
117 Justé et al. (2012), and our method yield accurate measurements in a previous study
118 (Fukui et al., 2012). Furthermore, we examined a partial sequence of the 16S rRNA gene of
119 L. taeanensis G5T, and confirmed that the sequence had 100 % similarity with that in database.
120 There were significant differences of > 10 mol% in the G+C contents between the three 121 strains (58.5-60.2 mol%) and the closest phylogenic neighbours (47.1-47.9 mol%),
122 which indicates the three strains belong to a different genus (Stackebrandt & Liesack,
123 1993). DNA-DNA hybridization relatedness among strains 0C-2-2T, 0C-17, and LNM-3
124 were 99.0-101.6 %, suggested that the three isolates belonged to the same species
125 (Wayne et al., 1987). On the other hand, strain 0C-2-2T was a different species from L.
126 taeanensis G5T (1.2-1.8 %) and H. balneolensis DSM 19091T (1.2-2.5 %).
127
128 Cell morphology and presences of a flagellum and a stalk were observed by a
129 transmission electron microscope (JEM-1200EX II, JEOL). Motility and Gram-stain
130 reaction (Merck) was ascertained by use of a light microscope (BX50, Olympus).
131 Colony morphology and pigmentation were observed on MA. Pigments of the cells
132 were extracted with acetone/methanol (7:2, v/v) (Biebl et al., 2005) and the absorption
133 spectrum between 350 and 900 nm was determined using a U-2000A spectrophotometer
134 (Hitachi). The following phenotypic tests were carried out using inoculated samples
135 over a period of 2 weeks. Growth under an anaerobic condition was tested on MA using
136 an Anaero Pack (Mitsubishi Gas Chemical). Temperature for growth was determined on
137 MA at 5-40 °C (at intervals of 5 °C). pH range for growth was determined in MB
138 adjusted to pH 4.0-10.0 (at intervals of one pH unit), using 1M NaOH and 1M HCl
139 solutions. After autoclaving, the pH values of MB were again measured and adjusted
140 with 1M NaOH and 1M HCl solutions. NaCl concentration for growth was tested in MB
141 prepared with various concentrations of 0.02, 0.5, 1-10 % (at intervals of 1 %, w/v)
142 NaCl (Alain et al., 2008). Phenotypic tests performed included the following: oxidase
143 (Oxidase reagent; bioMérieux) and catalase activities (3 % v/v H2O2), methyl red and
144 Voges-Proskauer reactions (Nissui Pharmaceutical), H2S production (Eiken Chemical), 145 hydrolysis of agar, alginate, DNA, starch, Tween 20, 40, and 80 (Sawabe et al., 1995),
146 and oxidation tests of various 10 % (w/v) carbon sources using OF basal medium
147 (Eiken Chemical). Other biochemical features were determined by using API 20NE,
148 API ZYM, and API 50CH (bioMérieux) following the manufacturer’s instructions
149 except that cells were suspended in 75 % artificial seawater (ASW; containing l-1
150 distilled water: 30 g NaCl, 0.7 g KCl, 5.3 g MgSO4. 7H2O, 10.8 g MgCl2. 6H2O,1.3 g
151 CaSO4. 2H2O). The test strips of API 20 and API ZYM were inoculated for 2 weeks and
152 1 day, respectively. Carbon utilization tests (API 50CH) were performed with basal
153 medium broth (Baumann et al., 1972) supplemented with 0.1 % (w/v) yeast extract for 3
154 weeks. Antibiotic susceptibility was determined using a disc diffusion method with
155 commercial disks (Eiken Chemical, Becton Dickinson) or filter paper disks impregnated
156 with different antibiotics. Antibiotic susceptibility as scored was positive when a
157 diameter of the inhibitory zone was above 10 mm (Alain et al., 2008). The phenotypic
158 characteristics of the three isolates are given in the species description and Table 2. The
159 main phenotypic features of the three isolates were rod-shaped cells, possession of a
160 polar flagellum or a prostheca (Fig. 2), Gram-negative staining reaction, and aerobic
161 growth, features that are frequently observed for members of the family
162 Hyphomonadaceae. The three isolates produced orange pigments with a maximum
163 absorption at 483-486 nm which was identical to a carotenoid (Biebl et al., 2005). The
164 absorption peak around 770 nm characterized by the presence of bacteriochlorophyll a
165 (Biebl et al., 2005) was not observed. The differences in phenotypic traits among the
166 three strains, L. taeanensis G5T, and H. balneolensis DSM 19091T are shown in Table 2.
167 In particular, eleven phenotypic features (mode of division, growth at 35 °C, nitrate
168 reduction, hydrolysis of gelatin, lipase and cysteine arylamidase activities, utilizations 169 of sucrose, starch, glycogen, and 2-keto-gluconate, and susceptibility to streptomycin)
170 were different between the three strains and the related species.
171
172 Isoprenoid quinones were extracted with chloroform/methanol (2:1, v/v) after
173 incubation in MB, and the type and length were determined by a HPLC method
174 according to the method described by Akagawa-Matsushita et al. (1992). Polar lipids
175 and methyl esters of cellular fatty acids (FAMEs) were extracted according to the
176 method described by Minnikin et al. (1984) and Ikemoto et al. (1978), respectively,
177 after the three isolates were cultured on MA at 20 °C for 10 days in stationary phase of
178 growth. The spots of the polar lipids on two-dimensional TLC were identified by
179 spraying with the appropriate detection reagents (Minnikin et al, 1984; Komagata &
180 Suzuki, 1987). The FAMEs was examined using a GC equipped with a flame-ionization
181 detecter (GC-2010; Shimadzu) and a capillary column (Omegawax 320; Supelco) and
182 were identified by comparing the retention times of a sample to those of a standard
183 (Supelco, USA). Furthermore, for the fatty acids (>1 % of the total), the FAMEs and the
184 DMOX derivatization (Luthria & Sprecher, 1993) were identified by a GC-MS (HP
185 5973N; Agilent Technologies). The predominant isoprenoid quinone of all isolates was
186 ubiquinone-10 (Q-10) which was observed in many members of the class
187 Alphaproteobacteria (Martens et al., 2006). TLC profiles of the polar lipids of the five
188 strains are shown in Fig. 3. The common polar lipids in all five isolates were
189 phosphatidylglycerol, glucuronopyranosyldiglyceride, mono-glycosyldiglyceride, three
190 unidentified phospholipids, and an unidentified glycolipid (GL1). Another unidentified
191 glycolipid (GL2) was detected in only L. taeanensis G5T. The presence of
192 phosphatidylglycerol, glucuronopyranosyldiglyceride, and mono-glycosyldiglyceride 193 are characteristic of members of the family Hyphomonadaceae (Alain et al., 2008). The
194 total cellular fatty acid compositions of all isolates are shown in Table 3. The major
195 fatty acids in the three strains were C18:1ω7c (34.9-71.6 %), which is a typical feature of
196 the class Alphaproteobacteria (Alain et al., 2008), followed by C18:1 2-OH, C18:0, and
197 C19:1ω8c. The compositions of C17:0, C18:1ω7c, C19:1ω8c, and C19:1 2-OH (>5 %) in strain
198 0C-17 were distinctly different from those in strains 0C-2-2T and LNM-3. In order to
199 ascertain these differences among three strains, we also examined FAMEs at early
200 period of incubation (8 days), but these results were similar to those in 10 days of
T 201 incubation. The three isolates specifically had C19:0, whereas either L. taeanensis G5 or
T 202 H. balneolensis DSM 19091 specifically had C18:2ω7c, 13c and C19:2ω7c, 14c. In
203 particular, the part of unsaturated fatty acids of nonadecanoate and 2-hydroxy fatty acids
204 were firstly detected in the family Hyphomonadaceae. The differences between this
205 study and previous studies (Alain et al., 2008; Jung et al., 2011) can be attributed to the
206 extraction method of FAMEs and culture temperatures.
207
208 In conclusion, strains 0C-2-2T, 0C-17, and LNM-3 are a novel genus and species
209 Algimonas porphyrae gen. nov., sp. nov on the basis of phylogenetic, genetic,
210 phenotypic, and chemotaxonomic features. In particular, A. porphyrae strains had
211 higher G+C contents (58.5-60.2 mol%) than related genera (47.1-47.9 mol%). The
212 eleven phenotypic features and the compositions of several fatty acids in the three
213 strains were also distinguished from those of the related genera.
214
215 Description of Algimonas gen. nov.
216 Algimonas (Al.gi.mo'nas. L. n. alga, seaweed; L. fem. n. monas, a monad, unit; N.L. 217 fem. n. Algimonas, a unit (bacterium) isolated from seaweed).
218 Cells are Gram-negative and straight to slight curved rods. Many cells are non-stalked
219 and possess a polar flagellum for motility. Some cells possess a prostheca.
220 Multiplication occurs by binary fission. Cells produce orange carotenoid pigments.
221 Bacteriochlorophyll a is not found. Aerobic condition and NaCl are required for growth.
222 The predominant isoprenoid quinone is Q-10. Polar lipids are phosphatidylglycerol,
223 glucuronopyranosyldiglyceride, mono-glycosyldiglyceride, three unidentified
224 phospholipids, and an unidentified glycolipid. The major cellular fatty acid is C18:1ω7c.
225 The genus belongs phylogenetically to the family Hyphomonadaceae in the class
226 Alphaproteobacteria. The G+C contents of the DNA are 58.5 to 60.2 mol%. The type
227 species of the genus is Algimonas porphyrae.
228
229 Description of Algimonas porphyrae sp. nov.
230 Algimonas porphyrae (por.phy'rae. N.L. gen. n. porphyrae, of Porphyra, isolated from
231 Porphyra yezoensis).
232 In addition to the description of the genus, the following properties are exhibited. The
233 cells are 2.13 μm long and 0.37 μm in diameter (n= 25) in MB after 5 days incubation.
234 Colonies on MA are circular, convex, entire, and 0.22 mm in diameter (n= 25) after 10
235 days incubation. Growth of the type strain occurs at 10-30 °C (optimum, 20 °C), pH
236 6.0-9.0 (optimum, pH 7.0-8.0), and 1-5 % of NaCl (optimum, 2-3 %). Oxidase is
237 negative and catalase is positive. Methyl red and Voges-Proskauer reactions are negative.
238 H2S and indole are not produced. Aesculin, Tween 20, 40, and 80 are hydrolysed, but
239 agar, alginate, DNA, gelatin, and starch are not. No acid is produced from carbohydrates.
240 Nitrate reduction activities are present. With API ZYM system, the type strain is 241 positive in alkaline phosphatase, esterase, esterase lipase, leucine arylamidase, valine
242 arylamidase, acid phosphatase, and naphthol-AS-BI-phosphohydrolase, but negative in
243 lipase, cystine arylamidase, trypsin, chymotrypsin, α-galactosidase, β-galactosidase,
244 β-glucuronidase, α-glucosidase, β-glucosidase, N-acetyl-β-glucosaminidase,
245 α-mannosidase, and α-fucosidase. With API 50CH system, the type strain utilizes
246 galactose, glucose, mannose, esculin ferric citrate, maltose, and starch are utilized, but
247 not glycerol, erythritol, D-arabinose, L-arabinose, ribose, D-xylose, L-xylose, adonitol,
248 methyl-β-D-xylopyranoside, fructose, sorbose, rhamnose, dulcitol, inositol, mannitol,
249 sorbitol, methyl-α-D-mannopyranoside, methyl-α-D-glucopyranoside,
250 N-acetyl-glucosamine, amygdalin, arbutin, salicin, cellobiose, lactose, melibiose,
251 sucrose, trehalose, inulin, melezitose, raffinose, glycogen, xylitol, gentiobiose,
252 D-turanose, D-lyxose, D-tagatose, D-fucose, L-fucose, D-arabitol, L-arabitol, gluconate,
253 2-keto-gluconate, or 5-keto-gluconate. The type strain is susceptible to (μg per disc
254 unless otherwise stated) ampicillin (10), carbenicillin (100), ciprofloxacin (5),
255 chloramphenicol (30), erythromycin (15), gentamicin (10), kanamycin (30), nalidixic
256 acid (30), neomycin (30), norfloxacin (10), novobiocin (30), penicillin G (10U),
257 rifampicin (5), streptomycin (10), and vancomycin (30) but resistance to polymyxin B
258 (300 U) and tetracycline (30).
259 The type strain is 0C-2-2T (LMG 26424T = NBRC 108216T) isolated from the red alga
260 Porphyra yezoensis. Strains 0C-17 (LMG 26425 = NBRC 108217) and LNM-3 (LMG
261 26426 = NBRC 108218) are reference strains.
262
263 Acknowledgements
264 We acknowledge Dr. J. P. Euzéby for support in the Latin etymologies of the genus 265 and species name. We are grateful to Professor C. O. Jeon of Chung-Ang
266 University for kindly providing Litorimonas taeanensis G5T. This study was supported
267 by Research Fellowships of the Japan Society for the Promotion of Science for Young
268 Scientists (Project No. 23·4176).
269
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377 Figure legends
378 Fig. 1. Neighbour-joining phylogenetic tree based on 16S rRNA gene sequences
379 showing the relationships among strains 0C-2-2T, 0C-17, and LNM-3, and
380 other type strains in the family Hyphomonadaceae. Bootstrap values (>50 %) based on
381 1000 replications are listed as percentages at branch nodes. Filled circles indicate that
382 the corresponding nodes were recovered by all algorithms. Open circles indicate that
383 the corresponding nodes were recovered by two algorithms. Escherichia coli ATCC
384 11775T was used as an outgroup. Bar, 0.02 % sequence divergence. 385
386 Fig. 2. Transmission electron micrographs of strain 0C-2-2T negatively stained. (a) A
387 cell with a polar flagellum. Bar = 0.5 μm. (b) A cell with a prostheca in a state of binary
388 fission. Bar = 0.5 μm.
389
390 Fig. 3. Polar lipid profiles of (a) strain 0C-2-2T, (b) strain 0C-17, (c) strain LNM-3, (d)
391 L. taeanensis G5T, and (e) H. balneolensis DSM 19091T. Two-dimensional TLC was
392 performed by chloroform-methanol-water 65: 25: 4 (vol/vol) as the first direction and
393 chloroform-acetic acid-methanol-water 80:15:12:4 (vol/vol) as the second direction. All
394 polar lipids were detected with 5 % ethanolic molybdophosphoric acid.
395 Phosphatidylglycerol (PG) was identified using a commercial standard (DOOSAN
396 Serdary Research Laboratories). PG, phosphatidylglycerol; GUDG,
397 glucuronopyranosyldiglyceride; MGDG, mono-glycosyldiglyceride; PL1-3, unidentified
398 phospholipid; GL1-2, unidentified glycolipid. 77 Hyphomonas hirschiana VP-5T (AF082794) 93 Hyphomonas rosenbergii VP-6 T (AF082795) 100 Hyphomonas neptunium ATCC 15444T (AF082798) Hyphomonas polymorpha DSM2665T (AJ227813) Hyphomonas adhaerens MHS-3T (AF082790) 100 100 Hyphomonas jannaschiana ATCC 33883T (AJ227814) Hyphomonas oceanitis SCH-89T (AF082797) 97 87 Hyphomonas johnsonii MHS-2T (AF082791) Ponticaulis koreensis GSW-23T (FM202497) T 68 87 Henriciella litoralis SD10 (FJ230835) T 100 Henriciella marina Iso4 (EF660760) 98 Henriciella aquimarina P38T (EU819081) 98 Hirschia baltica DSM 5838T (AJ421782) 100 Hirschia maritima GSW-2T (FM202386) Robiginitomaculum antarcticum IMCC3195T (EF495229) Hellea balneolensis 26III/A02/215T (AY576758) 100 Litorimonas taeanensis G5T (FJ230838) 85 Algimonas porphyrae 0C-2-2T (AB689189) 99 Algimonas porphyrae 0C-17 (AB689190) 100 Algimonas porphyrae LNM-3 (AB689191) 80 Oceanicaulis alexandrii C116-18T (AJ309862) Woodsholea maritima CM243T (AJ578476) 100 Maricaulis salignorans MCS 18T (AJ227806) 67 Maricaulis washingtonensis MCS 6T (AJ227804) 99 Maricaulis maris ATCC 15268T (AJ227802) T 81 Maricaulis parjimensis MCS 25 (AJ227808) 100 Maricaulis virginensis VC-5T (AJ301667) Escherichia coli ATCC 11775T (X80725)
0.02 Fig. 1. Fukui et al. (a) (b)
Fig. 2. Fukui et al. (a) PL1 MGDG (b)PL1 MGDG (c) PL1 MGDG PL2 PL2 PL2 GUDG GUDG GUDG PG PG PG
PL3 PL3 PL3 GL1 GL1 GL1
PL1 (d) MGDG (e) PL1 MGDG PL2 GUDG PL2 GUDG PG GL2 PG PL3
PL3 GL1 GL1
Fig. 3. Fukui et al. Table 1. DNA G+C contents and DNA-DNA hybridization values among novel strains and other related genera Strains DNA G+C content DNA-DNA reassociation value (%) with biotinylated DNA from: (mol%) A. porphyrae sp. nov. L. taeanensis H. balneolensis 0C-2-2T G5T DSM 19091T Algimonas porphyrae sp. nov.: 0C-2-2T 58.5 100.0 1.2 1.2 0C-17 59.0 101.6 1.5 1.9 LNM-3 60.2 99.0 2.0 2.4 Litorimonas taeanensis G5T 47.1 1.8 100.0 3.9 Hellea balneolensis DSM19091T 47.9 2.5 3.4 100.0 Table 2. Differential phenotypic characteristics among novel strains and other related species
Species/strains: 1, A. porphyrae sp. nov. 0C-2-2T; 2, A. porphyrae sp. nov. 0C-17; 3, A. porphyrae sp. nov. LNM- 3; 4, L. taeanensis G5T; 5, H. balneolensis DSM 19091T. +, Positive; -, negative; S, susceptible; R, resistant.
Characteristic 1 2 3 4 5 Mode of division Binary fission Binary fission Binary fission Budding Budding Pigmentation Orange Orange Orange Pale-orange Orange Growth at 5 °C---+- 10 °C+--+- 35 °C---++ Growth in pH 6.0, pH 9.0 + + + + - 0.5 % NaCl - - - + - 1 % NaCl + + + + - 5 % NaCl + - - + - 6-7 % NaCl - - - + - Oxidase activity - - - + - Hydrolysis of DNA - - - - + Tween 40, 80 + + + - + API 20NE Nitrate reduction + + + - - Hydrolysis of gelatin - - - + + Enzyme activity (API ZYM) Lipase - - - + + Cystine arylamidase - - - + + Acid phosphatase + - - + + Carbon utilization (API 50CH) Erythritol - - - - + Ribose - - - + - D-xylose - - - + - Adonitol - - - - + Galactose + - - - - Glucose + + + + - Mannose + - + - - Rhamnose - - - - + Dulcitol - - - - + Mannitol - - - - + Arbutin - - - - + Cellobiose - - - + - Maltose + + + - + Sucrose - - - + + Inulin - - - - + Starch + + + - - Glycogen - - - + + D-tagatose - - - - + L-arabitol - - - - + 2-keto-gluconate - - - + + Susceptibility to Carbenicillin (100 μg) S S R R S Kanamycin (30 μg) S S S R S Polymyxin B (300 U) R S R R R Streptomycin (10 μg) S S S R R Vancomycin (30 μg) S R S R S Table 3. Cellular fatty acid contents (%) of Algimonas porphyrae sp. nov. and other related genera Species/strains: 1, A. porphyrae sp. nov. 0C-2-2T; 2, A. porphyrae sp. nov. 0C-17; 3, A. porphyrae sp. nov. LNM-3; 4, L. taeanensis G5T; 5, H. balneolensis DSM 19091T. tr, trace amount (less than 0.5 %); ND, not detected. Fatty acid (%) 1 2 3 4 5
C17:0 0.6 6.1 0.5 4.4 3.5
C18:0 4.9 2.1 4.9 1.6 1.4
C19:0 1.4 0.5 1.2 ND ND
C16:1ω7c tr tr tr 1.1 ND
C17:1ω8c ND tr ND tr 1.3
C17:1ω7c tr 1.1 tr 5.1 0.9
C17:1ω6c tr 2.9 tr 6.2 2.8
C18:1ω7c 71.4 34.9 71.6 42.6 27.4
C18:2ω7c, 13c ND ND ND 1.4 2.6
C19:1ω9c tr 1.7 tr 0.7 1.3
C19:1ω8c 2.4 26.9 2.2 13.9 24.8
C19:1ω7c tr 3.8 tr 1.6 5.2
C19:2ω7c, 14c ND ND ND ND 5.7
C17:1 2-OH tr 1.3 tr 2.2 1.3
C18:1 2-OH 11.1 7.7 11.1 13.0 1.3
C19:1 2-OH tr 6.6 tr 1.1 1.3
iso-C18:1 2-OH tr tr ND tr 1.6
iso-C19:1 2-OH tr ND tr ND 8.8
iso-C20:1 2-OH tr tr ND ND 3.9
Unknown ECL 26.141 1.3 0.7 1.5 1.3 ND
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