1 Jiangella anatolica sp. nov. isolated from coastal lake soil
2 Hilal Ay1*, Imen Nouioui2, Lorena Carro2¥, Hans-Peter Klenk2, Demet Cetin3, José M.
3 Igual4, Nevzat Sahin1, Kamil Isik5
4 1H. Ay, N. Sahin
5 Department of Molecular Biology and Genetics, Faculty of Science and Arts, Ondokuz Mayis
6 University, Samsun, Turkey
7 *Corresponding author: [email protected], Tel: +90 362 312 1919, Fax: +90 362 457 6081
8 2I. Nouioui, L. Carro, H.-P. Klenk
9 School of Natural and Environmental Sciences, Newcastle University, Ridley Building 2,
10 Newcastle upon Tyne, NE1 7RU, UK
11 ¥Present address: Dpto. de Microbiología y Genética. Universidad de Salamanca, Salamanca,
12 37007, Spain.
13 3D. Cetin
14 Science Teaching Programme, Gazi Faculty of Education, Gazi University, 06500, Ankara,
15 Turkey
16 4J. M. Igual
17 Instituto de Recursos Naturales y Agrobiologia de Salamanca, Consejo Superior de
18 Investigaciones Cientificas (IRNASA-CSIC), Salamanca, Spain
19 5K. Isik
20 Department of Biology, Faculty of Science and Arts, Ondokuz Mayis University, Samsun,
21 Turkey
22 23
24 Abstract
25 A novel actinobacterial strain, designated GTF31T, was isolated from a coastal soil sample of
26 Gölcük Lake, a crater lake in southwest Anatolia, Turkey. The taxonomic position of the strain
27 was established using a polyphasic approach. Phylogenetic analyses based on 16S rRNA gene
28 sequences and showed that the strain is closely related to Jiangella gansuensis DSM 44835T
29 (99.4 %), Jiangella alba DSM 45237T (99.3 %) and Jiangella muralis DSM 45357T (99.2 %).
30 Optimal growth was observed at 28 °C and pH 7 – 8. Whole cell hydrolysates were found to
31 contain LL-DAP, glucose, mannose, rhamnose and ribose. The predominant menaquinone was
32 identified as MK-9(H4). The polar lipid profile was found to contain diphosphatidylglycerol,
33 phosphatidylglycerol, phosphatidylinositol, glycophospholipids and unidentified phospholipids.
34 The major fatty acids were identified as anteiso-C15:0, iso-C16:0 and anteiso-C17:0. The G+C
35 content of the type strain was determined to be 72.5 % and the size of the draft genome is 7.0
36 Mb. The calculated digital DDH values between strain GTF31T and the type strains of J.
37 gansuensis, J. alba, J. muralis and Jiangella alkaliphila ranged from 24.4 to 34.4% and ANI
38 values ranged between 81.0 – 87.9 %. Based upon the consensus of phenotypic and
39 phylogenetic analyses as well as whole genome comparisons, strain GTF31T (= DSM 100984T =
40 CECT 9378T) is proposed to represent the type strain of a novel species, Jiangella anatolica sp.
41 nov.
42 Keywords: Phylogenomic, Gölcük Lake, Jiangella, Polyphasic taxonomy
43 Introduction
44 The genus Jiangella was first proposed by Song et al. (2005) within the family
45 Nocardioidaceae. Subsequently, Tang et al. (2011) established the family Jiangellaceae to
46 accommodate the genera Jiangella (Song et al. 2005) and Haloactinopolyspora (Tang et al.
47 2011), mainly based on phylogenetic analysis of the phylum Actinobacteria. Currently, the 48 family Jiangellaceae includes the genera Jiangella, Haloactinopolyspora and the recently
49 described Phytoactinopolyspora (Li et al. 2015). The type genus Jiangella encompasses
50 aerobic, Gram-positive, filamentous actinomycetes with substrate mycelium which fragment
51 into short and elongated rods. Members of the genus are characterised by the presence of LL-
52 diaminopimelic acid in the cell wall peptidoglycan and MK-9(H4) as the major menaquinone.
53 None of the species described so far contain mycolic acids. The genus accommodates Jiangella
54 gansuensis YIM 002T (Song et al. 2005), Jiangella alba YIM 61503T (Qin et al. 2009),
55 Jiangella alkaliphila D8-87T (Lee 2008), Jiangella muralis 15-Je-017T (Kämpfer et al. 2011)
56 and Jiangella mangrovi 3SM4-07T (Suksaard et al. 2015) isolated from desert soil, surface
57 sterilised stem of Maytenus austroyunnanensis, cave soil, cellar wall and mangrove soil,
58 respectively.
59 Members of the phylum Actinobacteria are well-known for being prolific producers of many
60 bioactive secondary metabolites (Goodfellow and Fiedler 2010). Thus, selective isolation of
61 novel actinobacteria from unexplored habitats such as marine sediments (Carro et al. 2018) or
62 lakes (Ay et al. 2017; Ay et al. 2018) holds considerable importance for the discovery of novel
63 bioactive compounds. In a continuation of our investigations on actinobacterial diversity in
64 unexplored habitats a novel strain, GTF31T, was isolated from a coastal soil sample of Gölcük
65 Lake, a crater lake in southwest Anatolia (Turkey). Polyphasic characterisation of strain
66 GTF31T showed that the strain is a novel species of the genus Jiangella, for which the name
67 Jiangella anatolica sp nov is proposed.
68 Materials and methods
69 Isolation, maintenance and cultural conditions
70 Strain GTF31T was obtained from a coastal soil sample of Gölcük Lake, a freshwater lake, (37°
71 44’ 03.90’’ N; 30° 29’ 39.24’’ E), collected in September 2013. The strain was isolated on
72 International Streptomyces Project Medium 2 (ISP 2 medium) (Shirling and Gottlieb 1966) 73 supplemented with nystatin (50 µg/ml) and rifampicin (5 µg/ml) using a standard dilution-plate
74 technique. Briefly, the soil sample was suspended in ¼ strength Ringer’s solution (Oxoid) and
75 pretreated with wet heat at 60 ºC for 20 minutes. Then, the diluted suspensions (10-2 and 10-3)
76 were spread on ISP 2 medium. Inoculated plates were incubated at 28 ºC for two weeks. The
77 Jiangella isolate was purified and maintained on slopes of tryptone-yeast extract agar (DSM
78 medium No. 680) at 4 ºC and as glycerol stock solutions (25 % w/v) at – 20 ºC.
79 J. gansuensis JCM 13367T, J.alba JCM 16901T and J.muralis JCM 17970T were obtained from
80 Japan Collection of Microorganisms (JCM). These reference strains for phenotypic comparison
81 were grown and tested under identical conditions.
82 Cultural properties and phenotypic tests
83 Cultural and morphological characteristics were observed on ISP media 1 – 7 (International
84 Streptomyces Project, Shirling and Gottlieb (1966)), nutrient agar and tryptic soy agar (TSA;
85 Difco) after incubation at 28 °C for 14 days. The ISCC-NBS Colour Charts Standard Samples
86 No. 2106 (Kelly 1964) was used to determine the colours of aerial mycelium, substrate
87 mycelium and diffusible pigments. Micromorphological characteristics were observed by
88 scanning electron microscopy (JEOL JSM 6060) using cultures grown on glucose-yeast extract-
89 malt extract (DSMZ Medium No. 65) at 28 °C for 14 days. The temperature (4, 10, 20, 28, 37,
90 50 ºC) and pH range (7 – 12, at intervals of 1.0 pH unit) for growth were determined on ISP 2
91 medium (Shirling and Gottlieb 1966). Enzyme activity profile of the strain was established
92 using the commercial kit API ZYM (BioMerieux) following the manufacturer’s instructions.
93 Carbon source utilisation and chemical sensitivity properties were determined using GEN III
94 Microplates in an Omnilog device (BIOLOG Inc., Hayward, CA, USA). The GEN III
95 Microplates were inoculated with cells suspended in a viscous inoculation fluid (IF C) provided
96 by the manufacturer at 85–87 % of transmittance for strain GTF31T and the closely related type
97 strains. The strains were studied in two independent technical replicates. Data were exported
98 and analysed using the opm package for R (Vaas et al. 2013; Vaas et al. 2012). 99
100
101 Chemotaxonomy
102 Biomass for chemotaxonomic analyses was obtained by cultivation in shake flasks at 28 °C for
103 14 days on a rotary shaker (150 rpm) using glucose-yeast extract-malt extract (DSMZ Medium
104 No. 65). Biomass was harvested by centrifugation at 9000 rpm, washed several times with
105 sterile distilled water and then freeze-dried. Whole cell amino acids and sugars were extracted
106 following a standard protocol described by Lechevalier and Lechevalier (1970) and analysed by
107 thin layer chromatography (TLC) according to Staneck and Roberts (1974). Polar lipids were
108 extracted and analysed by two-dimensional TLC following Minnikin et al. (1984) using
109 modifications of Kroppenstedt and Goodfellow (2006). Menaquinones were extracted from
110 freeze-dried biomass and purified according to Collins (1985). The extracted menaquinones
111 were analysed using a high performance liquid chromatography (HPLC; LC-10AS, Shimadzu
112 63 Co.) fitted with a reverse-phase C18 column (150 x 4.6 mm, 5 µm particle size, RP-18-
113 Lichrosorb column, Penomenex UK) at 270 nm. Fatty acid methyl esters were extracted from
114 fresh cultures grown on TSA at 28 °C for 10 days and analysed using Microbial Identification
115 System (MIDI) Sherlock software version 6.1 and the library RTSBA6 according to the
116 technical instructions provided by this system (Sasser 1990).
117 Phylogeny
118 Genomic DNA was extracted as described by Chun and Goodfellow (1995) and PCR
119 amplification of the 16S rRNA gene achieved using the universal primers 27F (5′–
120 AGAGTTTGATC(AC)TGGCTCAG–3′) and 1492R (5′–
121 ACGG(CT)TACCTTGTTACGACTT–3′) (Weisburg et al. 1991). The purified PCR products
122 were sequenced using an ABI PRISM 3730 XL automatic sequencer and an almost complete
123 16S rRNA gene sequence (1491 bp) was deposited in the GenBank data library and 124 corresponding sequences of the type strains of the genus Jiangella were retrieved by using the
125 EzBioCloud server (Yoon et al. 2017). A multiple sequence alignment was generated with
126 MUSCLE program (Edgar 2004). A Neighbour Joining tree (Saitou and Nei 1987) was
127 constructed in MEGA 6 (Tamura et al. 2013) using the model of Jukes and Cantor (Jukes and
128 Cantor 1969). Phylogenetic analyses for maximum likelihood (ML) and maximum parsimony
129 (MP) methods were conducted by the GGDC web server using the DSMZ phylogenomics
130 pipeline (Meier-Kolthoff et al. 2014) adapted to single genes after pairwise sequence similarities
131 were calculated according to Meier-Kolthoff et al. (2013a) via the GGDC web server (Meier-
132 Kolthoff et al. 2013b) available at http://ggdc.dsmz.de/. The ML tree was inferred from the
133 alignment with RAxML (Stamatakis 2014) using rapid bootstrapping in conjunction with the
134 autoMRE criterion (Pattengale et al. 2010) and the MP tree was inferred from the alignment
135 with TNT (Goloboff et al. 2008) using 1000 bootstraps in conjunction with tree-bisection-and-
136 reconnection branch swapping and ten random sequence additional replicates. The sequences
137 were checked for compositional bias using the X2 test as implemented in PAUP* (Swofford
138 2002).
139 The genome of strain GTF31T was sequenced using the Illumina HiSeq 2500 next generation
140 sequencing platform with a 250-bp paired end protocol (MicrobesNG, Birmingham, United
141 Kingdom) and the draft genome sequence was submitted to NCBI under the accession number
142 POTW01000000. The genome sequence was annotated on the Rapid Annotations Using
143 Subsystems Technology (RAST) server (Aziz et al. 2008). A phylogenomic tree was built on
144 the PATRIC web server (https://patricbrc.org/) (Wattam et al. 2016) using FastTree algorithm
145 (Price et al. 2010). The digital DNA-DNA hybridization (dDDH) similarities between the draft
146 genome sequence of strain GTF31T and publicly available genome sequence data of its close
147 phylogenetic neighbours were determined using formula 2 of the GGDC web server (Meier-
148 Kolthoff et al. 2013b). In addition, the level of pairwise genome-based similarities was 149 evaluated based on the average nucleotide identity (ANI) value determined by using online ANI
150 calculator described by Yoon et al. (2017) available at https://www.ezbiocloud.net/tools/ani.
151 Results and discussion
152 Strain GTF31T was found to grow well on ISP 1, ISP 2, ISP 3, ISP 6, nutrient agar and TSA
153 while moderate growth was observed on ISP 5 and ISP7 media. The strain could not grow on
154 ISP 4 medium and no diffusible pigment was observed on any media tested nor did it produce
155 melanin pigment. The substrate mycelium was observed to fragment into elongated rods (2.1
156 µm x 0.3 µm) while aerial mycelium and spore formation could not be observed during a 14-
157 day incubation period on any media tested (Fig 1). The strain was found to be able to grow at
158 temperature and pH range of 20 – 37 °C (optimum at 28 °C) and 6 – 12 (optimum at pH 7 – 8),
159 respectively, and in the presence of 1% NaCl. Strain GTF31T can oxidise various compounds
160 including acetic acid, acetoacetic acid, D-arabitol, D-cellobiose, dextrin, gelatin, β-gentiobiose,
161 D-glucose, N-acetyl-D-glucosamine, β-methyl-D-glucoside, glycerol, L-lactic acid, D-maltose,
162 D-mannitol, N-acetyl-β-D-mannosamine, pectin, D-salicin, sucrose, D-trehalose and turanose.
163 The strain is able to respire in presence of aztreonam, lincomycin, nalidixic acid, potassium
164 tellurite, rifamycin SV, 1% sodium lactate and troleandomycin whereas it could not tolerate
165 fusidic acid, guanidine hydrochloride, lithium chloride, minocycline, niaproof, sodium bromate
166 and tetrazolium blue (Table 1). Allantoin and urea hydrolysis and nitrate reduction tests were
167 found to be positive while arbutin hydrolysis is negative. In addition, strain GTF31T was found
168 to produce alkaline phosphatase, esterase (C4), esterase lipase (C8), lipase (C14), cystine
169 arylamidase, trypsin, α-chymotrypsin, acid phosphatase, naphthol-AS-BI-phosphohydrolase, α-
170 galactosidase, β-galactosidase, β-glucuronidase, α-glucosidase, β-glucosidase, N-acetyl-β-
171 glucosaminidase, α-mannosidase, α-fucosidase as shown by API ZYM (BioMerieux) tests.
172 Chemotaxonomic characteristics of strain GTF31T are consistent with those of members of the
173 genus Jiangella such as LL-diaminopimelic acid and glucose in whole cell hydrolysates. The
174 strain was also found to contain mannose, rhamnose and ribose. The polar lipid profile was 175 found to contain diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, four
176 glycophospholipids and three unidentified phospholipids (Fig S1). The predominant
177 menaquinone was determined to be MK-9(H4). The major cellular fatty acids were identified as
178 anteiso-C15:0 (28.7%), iso-C16:0 (16.9%) and anteiso-C17:0 (16.4%) (Table S1).
179 An almost complete 16S rRNA gene (1491 bp) was obtained and analysed on the EzBiocloud
180 server and also on the GGDC web server. Phylogenetic analyses of 16S rRNA gene sequences
181 showed that strain GTF31T is a member of the genus Jiangella. The strain is closely related to J.
182 gansuensis DSM 44835T, J. alba DSM 45237T, J. muralis DSM 45357T, J. mangrovi 3SM4-07T
183 and J. alkaliphila DSM 45079T with 16S rRNA gene sequence similarities of 99.4%, 99.3%,
184 99.2%, 99.1% and 98.8%, respectively. Strain GTF31T was found to form a distinct branch at
185 the periphery of Jiangella 16S rRNA gene trees constructed by both the neighbour joining (Fig
186 2) and maximum likelihood (Fig S2) tree-making algorithms.
187 The draft genome sequence of strain GTF31T has been deposited in GenBank with accession
188 number of POTW00000000 and annotated with the RAST server. The draft genome is
189 composed of 256 contigs with N50 value of 54,053. The numbers of coding sequences and
190 RNAs are 6444 and 51, respectively. The total genome size is 7 Mb and the G+C content is
191 72.5%. In a phylogenomic tree constructed using the PATRIC web server, strain GTF31T was
192 found to form a distinct cluster with other members of the genus Jiangella (Fig 3). The
193 calculated digital DDH values between strain GTF31T and the type strains of J. gansuensis, J.
194 alba, J. muralis and J. alkaliphila ranged from 24.4 to 34.4%, values which are well below the
195 proposed threshold of 70% for prokaryotic species delineation (Wayne et al. 1987). In addition
196 to low DDH values, ANI values calculated on the EzBioCloud web server (Table S2) ranged
197 between 81.0 – 87.9 % (values below the threshold of 95-96 %; Richter and Rosselló-Móra
198 2009), which confirmed the differentiation of strain GTF31T from its phylogenetic neighbours.
199 On the basis of phylogenetic, chemotaxonomic and morphological analyses, strain GTF31T
200 shows characteristics typical of members of the genus Jiangella but differs from previously 201 described species. Therefore, it is clear that strain GTF31T represents a novel member of the
202 genus Jiangella for which Jiangella anatolica sp nov. is proposed.
203
204 Description of Jiangella anatolica sp. nov.
205 Jiangella anatolica (a.na.to’li.ca. N.L. fem. adj. anatolica, referring the place from which the
206 type strain was isolated).
207 Aerobic, Gram-positive actinomycete that forms mycelia fragmenting into elongated rods. The
208 substrate mycelium is cream or pebble grey on ISP 1, ISP 2, ISP 3, ISP 5, ISP 6, ISP 7, nutrient
209 agar and TSA media. Shows good growth on ISP 1 – 3, ISP 6, nutrient agar and TSA media
210 while moderate on ISP 5 and ISP 7 media. Optimal growth occurs at 28 °C and pH 7 – 8. Whole
211 cell hydrolysates contain LL-diaminopimelic acid, glucose, mannose, rhamnose and ribose. The
212 predominant menaquinone is MK-9(H4). The polar lipid profile consists of
213 diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, glycophospholipids and
214 unidentified phospholipids. Major fatty acids are anteiso-C15:0, iso-C16:0 and anteiso-C17:0. The
215 G+C content of the type strain is 72.5 % and the size of the draft genome of the type strain is 7.0
216 Mb.
217 The type strain GTF31T (= DSM 100984T = CECT 9378T) was isolated from a coastal soil
218 sample of Gölcük Lake, Isparta, Turkey. The GenBank/EMBL/DDBJ accession numbers of the
219 16S rRNA gene sequence and draft genome sequence of strain GTF31T are KU255546 and
220 POTW01000000, respectively.
221 Acknowledgements
222 HA is grateful for the Scientific and Technological Research Council of Turkey (TÜBİTAK) for
223 the PhD scholarship. IN and LC are grateful for Newcastle University for the postdoctoral
224 fellowship. 225
226
227
228 Author contributions
229 HA, NS and KI designed the study. HA, IN and LC conducted physiological experiments. HPK,
230 HA, IN, JMI designed chemotaxonomic analyses. DC carried out scanning electron microscopy
231 analysis.
232 Compliance with ethical standards
233 Conflict of interest
234 The authors declare that they have no conflict of interest.
235 Ethical approval
236 This article does not contain any studies with human participants and/or animals performed by
237 any of the authors. The formal consent is not required in this study.
238 Supplementary material
239 Table S1 Fatty acid composition of strain GTF31T, Jiangella gansuensis JCM 13367T, J. alba
240 JCM 16901T, J. muralis JCM 17970T
241 Table S2 Genome sequence comparison between strain GTF31T and closely related type strains
242 of the genus Jiangella. Genome accession numbers are given in brackets
243 Fig. S1 Polar lipid profile of strain GTF31T. Key: DPG, diphosphatidylglycerol; PG,
244 phosphatidylglycerol; PI, phosphatidylinositol; GPL1-4, unidentified glycophospholipids; PL1-
245 3, unidentified phospholipid 246 Fig. S2 Maximum likelihood phylogenetic tree based on 16S rRNA gene sequences showing
247 relationships between strain GTF31T and closely related type strains. The tree was inferred
248 under the GTR+GAMMA model. The numbers above the branches are support values when
249 larger than 60% from maximum likelihood (left) and maximum parsimony (right)
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Table 1 Phenotypic characteristics that distinguish strain GTF31T from closely related type strains of the genus Jiangella. J. gansuensis JCM 13367T, J.alba JCM 16901T, J.muralis JCM
17970T. All data was obtained from the present study and the strains involved in phenotypic comparison grown and tested under identical conditions
GTF31T JCM 13367T JCM 16901T JCM 17970T Respiration in presence of
D-Arabitol + + + -
γ-Amino-n-Butyric Acid - - + -
β-Hydroxy-Butyric Acid - + - -
Citric Acid + - + -
D-Fructose - + + +
D-Fucose + - - +
D-Galactose - + - -
N-Acetyl-D-Galactosamine - + + +
D-Galacturonic Acid - - + -
L-Galactonic Acid-γ-Lactone - - + -
D-Gluconic Acid - - + -
3-O-Methyl-D-Glucose - + - -
L-Glutamic Acid + + + -
α-Keto-Glutaric Acid - - + -
Inosine + - + + myo-Inositol - + - -
D-Malic Acid - - - +
L-Malic Acid - - + -
D-Mannitol + - + -
D-Mannose + - - +
D-Melibiose + - - -
D-Lactic Acid Methyl Ester - - + -
α-D-Lactose + + + +
Pectin + + + +
D-Raffinose + - + + D-Serine - + - -
D-Sorbitol + - + +
Stachyose - - + +
Resistance to
4% NaCl - + + +
8% NaCl - + - +
Butyric Acid + + - +
Fusidic Acid - + - -
Guanidine Hydrochloride - - + +
Lithium Chloride - + + +
Minocycline - - - +
Mucic Acid - - + -
Propionic Acid + - - +
Quinic Acid + - - -
Sodium Bromate - + + +
Tetrazolium Violet + - + +
Tetrazolium Blue - + + +
Vancomycin + - - -
+ positive reaction, - negative reaction. All strains could respirate in presence of dextrin, D- maltose, D-trehalose, D-cellobiose, β-gentiobiose, sucrose, turanose, α-D-lactose, β-methyl-D- glucoside, D-salicin, N-acetyl-D-glucosamine, N-acetyl-β-D-mannosamine, D-glucose, L- fucose, L-rhamnose, glycerol, gelatin, pectin, L-lactic acid, Tween 40, acetoacetic acid, acetic acid but not N-acetyl-neuraminic acid, D-glucose-6-phosphate, D-fructose-6-phosphate, D- aspartic acid, D-serine, L-alanine, L-arginine, L-aspartic Acid, L-histidine, L-pyroglutamic acid,
L-serine, D-glucuronic acid, glucuronamide, D-saccharic acid, p-hydroxy-phenylacetic acid, methyl pyruvate, bromo-succinic acid, α-hydroxy-butyric acid, α-keto-butyric acid, sodium formate. All strains could tolerate NaCl up to 1%, troleandomycin, rifamycin SV, lincomycin, nalidixic acid, potassium tellurite, aztreonam but not niaproof. None of the strains could grow at pH 5.
Table S1 Fatty acids composition of strain GTF31T, Jiangella gansuensis JCM 13367T, J. alba
JCM 16901T, J. muralis JCM 17970T. All data was obtained from the present study and the strains were grown under identical conditions
Fatty acid GTF31T JCM 13367T JCM 16901T JCM 17970T
Saturated fatty acids
C12:0 - 1.4 - -
C16:0 - 2.2 1.5 -
C17:0 5.2 1.4 2.7 6.2
C18:0 - 2.8 4.2 -
C17:0 10-methyl - - - 1.7
Unsaturated fatty acids
C17:1 ω8c 2.1 3.2 9.8 2.7
C18:1 ω9c - 3.1 0.8 - iso-C15:1 G - 1.8 - - iso-C16:1 G 3.1 4.1 2.4 1.7 iso-C17:1 ω9c - 1.9 0.9 - anteiso-C15:1 A 1.1 3.3 - - anteiso-C17:1 1.1 4.9 - -
ω9c anteiso-C17:1 A - 1.7 0.4 -
Branched fatty acids iso-C14:0 3.4 2.2 7.0 7.6 iso-C15:0 5.4 9.7 11.9 5.7 iso-C16:0 16.9 7.8 15.5 21.4 iso-C17:0 3.0 3.6 1.9 1.5 iso-C18:0 2.0 - - 1.7 anteiso-C15:0 28.7 28.2 31.4 26.0 anteiso-C17:0 16.4 13.6 11.3 10.1
Hydroxy fatty acids iso-C14:0 3-OH - - - 2.0
C15:0 2-OH 2.7 2.5 2.0 - iso-C15:0 3-OH 1.0 1.8 - -
Table S2 Genome sequence comparison between strain GTF31T and closely related type strains of the genus Jiangella. Genome accession numbers are given in brackets
Strain GTF31T (POTW01000000)
ANI (%) dDDH (%)
J. gansuensis DSM 44835T (AZXT01000000) 81.0 24.4
J. alba DSM 45237T (FNUC00000000) 87.9 34.4
J. muralis DSM 45357T (LFXL01000000) 87.9 34.4
J. alkaliphila DSM 45079T (LT629791) 87.7 33.9
Fig. 1 Scanning electron micrograph of strain GTF31T grown on glucose-yeast extract-malt extract agar (DSMZ Medium No. 65) at 28 °C for 14 days
Fig. 2 Neighbour joining phylogenetic tree based on 16S rRNA gene sequences of strain
GTF31T and related type species. Numbers at the nodes represent bootstrap values. Only bootstrap values more than ≥50% are shown. Bar indicates 0.01 substitutions per site
Fig. 3 Phylogenomic tree showing relationship between strain GTF31T and closely related species of the genus Jiangella as well as other members of the family Jiangellaceae was built on PATRIC web server (https://patricbrc.org/) (Wattam et al. 2017) using FastTree algorithm
(Price et al. 2010). Numbers at the nodes indicate support values for confidence of subtrees within the larger tree
Fig. S1 Polar lipid profile of strain GTF31T. Key: DPG, diphosphatidylglycerol; PG, phosphatidylglycerol; PI, phosphatidylinositol; GPL1-4, unidentified glycophospholipids; PL1-
3, unidentified phospholipids
ig. S2 Maximum likelihood phylogenetic tree based on 16S rRNA gene sequences showing relationships between strain GTF31T and closely related type strains. The tree was inferred under the GTR+GAMMA model. The numbers above the branches are support values when larger than 60% from maximum likelihood (left) and maximum parsimony (right) bootstrapping. The branches are scaled in terms of the expected number of substitutions per site
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