maribellus comscasis sp. nov., isolated from the deep-sea cold seep · 2021. 1. 6. · abstract a...

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Maribellus comscasis sp. nov., isolated from the deep-sea cold seep 1

Rikuan Zheng1,2,3,4, Chaomin Sun1,2,4* 2

1CAS Key Laboratory of Experimental Marine Biology & Center of Deep Sea 3

Research, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China 4

2Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory 5

for Marine Science and Technology, Qingdao, China 6

3College of Earth Science, University of Chinese Academy of Sciences, Beijing, 7

China 8

4Center of Ocean Mega-Science, Chinese Academy of Sciences, Qingdao, China 9

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* Corresponding author 11

Chaomin Sun Tel.: +86 532 82898857; fax: +86 532 82898857. 12

E-mail address: [email protected] 13

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Category: New taxon in Bacteroidetes 15

Running title: A novel Bacteroidetes bacterium 16

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The NCBI GenBank accession numbers for the 16S rRNA gene sequence and 18

whole-genome sequence (WGS) of strain WC007T are MN096653 and CP046401, 19

respectively. 20

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(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. The copyright holder for this preprintthis version posted January 6, 2021. ; https://doi.org/10.1101/2021.01.06.425552doi: bioRxiv preprint

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ABSTRACT 27

A facultatively anaerobic, Gram-stain-negative, non-motile, curved rod-shaped 28

bacterium, designated WC007T, was isolated from the deep-sea cold seep, P. R. 29

China. Strain WC007T was found to grow at temperatures from 28 to 37 °C (optimum, 30

30 °C), at pH values between pH 6.0 and 8.0 (optimum, pH 7.0) and in 0-5.0% (w/v) 31

NaCl (optimum, 1.0%). The major fatty acids (>10.0%) were iso-C15:0, C16:0, summed 32

feature 3 and summed feature 8. The major isoprenoid quinone was MK-7. 33

Predominant polar lipids were phosphatidylethanolamine, one unidentified 34

phospholipid, one unidentified aminolipid and one unidentified lipid. The G+C 35

content of the genomic DNA was 38.38%. The average nucleotide identity (ANIb and 36

ANIm), amino acid identity (AAI), the tetranucleotide signatures (Tetra) and in silico 37

DNA-DNA hybridization (isDDH) similarities between the genome sequences of 38

isolate WC007T and Maribellus luteus XSD2T were 70.11%, 84.94%, 71.0%, 0.92022 39

and 20.40%, respectively, indicating that strain WC007T was distinguished from M. 40

luteus. Phylogenetic analysis based on 16S rRNA gene sequences placed strain 41

WC007T within the genus Maribellus and showed the highest similarity to strain 42

XSD2T (95.70%). In combination of the results of phylogenetic analysis and 43

phenotypic and chemotaxonomic data, strain WC007T was considered to represent a 44

novel species of the genus Maribellus, for which the name Maribellus comscasis sp. 45

nov. is proposed. The type strain is WC007T (=KCTC 25169T = MCCC 1K04777T). 46

The available of the genome sequence of strain WC007T would be helpful in 47

understanding the degradation mechanism of difficult-to-degrade polysaccharides. 48

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Keywords 51

Maribellus comscasis, 16S rRNA gene sequence, polysaccharides, deep-sea cold seep 52

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The family Prolixibacteraceae was originally described and classified under the order 55

Bacteroidales [1], class Bacteroidia, phylum Bacteroidetes, and the genus 56

Prolixibacter is the type genus [2]. After that, the families Prolixibacteraceae, 57

Marinilabiliaceae and Marinifilaceae were transferred to the order Marinilabiliales in 58

2016 [3]. At the time of writing, 11 genera including Aquipluma, Meniscus, 59

Puteibacter, Sunxiuqinia, Prolixibacter, Mangrovibacterium, Draconibacterium, 60

Mariniphaga, Tangfeifania, Roseimarinus and Maribellus had been identified within 61

the family Prolixibacteraceae [4-6]. Members of the family Prolixibacteraceae were 62

isolated from various habitats, such as coastal sediment [7, 8], crude oil [9], mangrove 63

sediment [1], freshwater lake [5], and marine sediments [10]. The new genus 64

Maribellus within the family Prolixibacteraceae was proposed recently [4], and 65

Maribellus luteus XSD2T, isolated from coastal seawater, was the only strain in the 66

genus Maribellus. The predominant respiratory quinone of strain XSD2T was MK-7, 67

which is the most frequently identified respiratory quinone in bacteria belonging to 68

the family Prolixibacteraceae [4]. 69

The phylum Bacteroidetes, specialized on polysaccharide degradation, was the 70

most abundant group of bacteria in the ocean after Proteobacteria and Cyanobacteria 71

[11]. Marine Bacteroidetes were commonly assumed to have a key role in degrading 72

phytoplankton polysaccharides [12], which had a great number and diversity of 73

carbohydrate-active enzymes (CAZymes) [13]. The CAZymes are categorized into 74

families of glycoside hydrolases (GHs), glycoside transferases (GTs), 75

carbohydrate-binding modules (CBMs), carbohydrate esterases (CEs), polysaccharide 76

lyases (PLs), sulfatases (targeting sulfated polysaccharides) plus a range of auxiliary 77

enzymes [12, 14, 15]. And the components of Bacteroidetes for the degradation of 78

polysaccharides was often encoded in distinct polysaccharide utilization loci (PULs) 79

[16], which were strictly regulated by the gene clusters that encode CAZymes and 80

protein ensembles required for the degradation of complicated carbohydrates. Besides 81

the various substrate-specific CAZymes genes, Bacteroidetes PULs also contain genes 82


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encoding SusCD-like proteins that are extracellular lipoproteins and integral 83

membrane beta-barrels termed TonB-dependent transporters [17]. In addition, PULs 84

of marine Bacteroidetes are frequently found many sulfatases [15, 18-21], because 85

abundant polysaccharides from marine algae were often sulfated. So far most 86

functional studies of Bacteroides polysaccharide degradation have been conducted in 87

human gut Bacteroides [22], however, only a few studies have investigated 88

polysaccharide degradation by marine Bacteroides, especially the deep-sea variety 89

[23]. 90

In this study, a facultatively anaerobic strain WC007T, belonging to the 91

Maribellus genus, was isolated from deep-sea sediment at a depth of 1,146 m in the 92

cold seep (22º 06' 58.598'' N 119º 17' 07.322'' E) as described previously [24], P. R. 93

China. Strain WC007T was isolated from an enrichment medium containing (per liter 94

of seawater): 1.0 g NaHCO3, 1.0 g CH3COONa, 1.0 g NH4Cl, 0.5 g KH2PO4, 0.2 g 95

MgSO4.7H2O, 1.0 g polysaccharide (cellulose, pectin and xylan), 1.0 mL 0.1 % (w/v) 96

resazurin, 0.7 g cysteine hydrochloride (pH 7.0) at atmospheric pressure and the 97

medium was prepared anaerobically as previously described [25]. The cultures were 98

repeatedly purified by using the Hungate roll-tube method in the medium containing 99

1.5% (w/v) agar. After incubation for 7 days, several colonies were picked by 100

sterilized bamboo skewers and then harvested and cultured in the liquid medium. The 101

process of isolation was repeated several times until the isolates were deemed to be 102

axenic. The purity of this isolate was confirmed routinely by transmission electron 103

microscopy (TEM) and by repeated sequencing of the 16S rRNA gene. Then the 104

single colony was transferred to a new medium (ORG) for further culture. The ORG 105

medium contained (L−1): 1.0 g NaHCO3, 1.0 g NH4Cl, 1.0 g CH3COONa, 0.2 g 106

MgSO4.7H2O, 0.5 g KH2PO4, 1.0 g yeast extract, 1.0 g peptone, 0.7 g cysteine 107

hydrochloride, 1 mL 0.1% (w/v) resazurin; the pH was adjusted to 7.0 [26]. The 108

isolate was cultured in ORG broth and maintained at -80 °C as a suspension in ORG 109

supplemented with glycerol (20 %, v/v). 110


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Genomic DNA was extracted from strain WC007T cultured for 7 d at 30 °C using 111

a bacteria genomic DNA kit (Takara, Japan). The whole genome sequencing (WGS) 112

of strain WC007T was performed on the nanopore sequencing technology platform 113

with the Oxford Nanopore MinION (Oxford, UK) and Illumina MiSeq sequencing 114

platform (San Diego, USA). The experimental process was implemented in 115

accordance with the standard protocol provided by Oxford Nanopore Technologies 116

(Oxford, UK). And the library building includes the following four steps: (1) High 117

quality genomic DNA was extracted, and then Nanodrop, Qubit and 0.35 % agarose 118

gel electrophoresis were used for purity, concentration and integrity inspection, 119

respectively; (2) The large fragments of DNA were recovered by the BluePippin 120

automatic nucleic acid recovery system; (3) Library construction was conducted by 121

using the SQK-LSK109 connection Kit (Japan), which including DNA damage repair 122

and terminal repair, magnetic bead purification, ligation of sequencing adapters and 123

magnetic bead purification. (4) After the library construction, computer sequencing 124

will be performed and Canu V1.5 software was used to assemble the filtered subreads 125

[27]. Finally, Pilon software was used to correct the assembled genome with 126

second-generation data to obtain the final genome with higher accuracy [28]. 127

The whole genome of strain WC007T had been deposited at GenBank under the 128

accession number CP046401. The genome size of strain WC007T was 7,811,310 bp 129

with a DNA G+C content of 38.38%. The number of contigs was 1, the total of N50 130

was 7,811,310 and the sequencing depth was 50.0×. Annotation of the genome of 131

strain WC007T consisted of 6,176 coding sequences that included 54 RNA genes (6 132

rRNA genes, 45 tRNA genes and 3 other ncRNAs). And we checked the authenticity 133

of the genome using the QUAST-5.0.2 software. The results showed that the genome 134

assembly quality of WC007T was high. In the genome of WC007T, genes encoding 135

SusC, SusD, CAZymes and sulfatase were ubiquitously distributed, strongly 136

indicating it possesses potentials for polysaccharide degradation. As measures of 137

relatedness between strain WC007T and closely related strains, the genome 138


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relatedness values were calculated by several approaches: Average Nucleotide 139

Identity (ANI) based on the MUMMER ultra-rapid aligning tool (ANIm), ANI based 140

on the BLASTN algorithm (ANIb), the tetranucleotide signatures (Tetra), and in silico 141

DNA–DNA similarity. ANIm, ANIb, and Tetra frequencies were calculated, using 142

JSpecies WS (http://jspecies.ribohost.com/jspeciesws/) [29]. The recommended 143

species criterion cut-offs were used; 95% for the ANIb and ANIm and 0.99 for the 144

Tetra signature [30]. The amino acid identity (AAI) values were calculated by 145

AAI-profiler (http://ekhidna2.biocenter.helsinki.fi/AAI/) [31]. The in silico 146

DNA-DNA similarity (isDDH) values were calculated by the Genome-to-Genome 147

Distance Calculator (GGDC) (http://ggdc.dsmz.de/) [32]. The isDDH results were 148

based on the recommended formula 2, which is independent of genome size and, thus, 149

is robust when using whole-genome sequences. 150

From ANI calculations, it could be observed that the comparison between strain 151

WC007T and three closely relatives were noticeably lower (ANIb: 70.11-70.51% and 152

ANIm: 83.71-84.94%) than the 95% cut-off proposed for bacterial species delineation. 153

The AAI values of 71.0-71.7% between strain WC007T and closely relatives were 154

obtained, which were also far below the 95% cut-off value generally recommended 155

for species differentiation. The Tetra values were also lower (0.92022-0.9496) than 156

the 0.99 cut-off proposed for bacterial species differentiation. Finally, the isDDH 157

values were significantly lower (18.40-20.40%) than the 70% cut-off value generally 158

recommended for species differentiation. The related genome comparison data were 159

listed in the Table S1. 160

Genomic DNA extraction and PCR amplification of the 16S rRNA gene of strain 161

WC007T were carried out as described by Hetharua et al [33]. The PCR product was 162

cloned into the vector pMD-19T (TaKaRa, Japan), sequenced and then compared to 163

the 16S rRNA gene sequence extracted from the genome. The results exhibited 99.9 % 164

sequence similarity. The whole full-length 16S rRNA gene sequence (1,516 bp) of 165

WC007T was obtained from the genome, which had been deposited in the GenBank 166


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database (accession number MN096653). Moreover, the 16S rRNA gene sequences of 167

related taxa were obtained from NCBI (www.ncbi.nlm.nih.gov/). A sequence 168

similarity calculation using the NCBI server indicated the closest relatives of strain 169

WC007T were Maribellus luteus XSD2T (95.70%), Mariniphaga sediminis SY21T 170

(93.44%) and Draconibacterium sediminis MCCC 1A00734T (92.99%). Phylogenetic 171

analysis was performed using the software MEGA version 6.0 [34]. Phylogenetic 172

trees were constructed by the neighbor-joining algorithm [35], maximum Likelihood 173

[36] and minimum-evolution methods [37]. The numbers above or below the branches 174

are bootstrap values based on 1,000 replicates. Phylogenetic analyses based on the 175

16S rRNA and genome sequences showed strain WC007T belonged to the genus 176

Maribellus and formed an independent phyletic line with the type strain Maribellus 177

luteus XSD2T (Fig. 1 and Fig. S1). Therefore, we propose strain WC007T to be 178

classified as the type strain of a novel species in the genus Maribellus, for which the 179

name Maribellus comscasis sp. nov. is proposed. On the basis of the phylogenetic 180

results, Maribellus luteus XSD2T (=MCCC 1H00347T) was selected as the closest 181

recognized neighbor of strain WC007T and was used as a reference strain in most of 182

the subsequent phenotypic tests. 183

To observe the morphological characteristics of M. comscasis WC007T, cells were 184

examined using transmission electronic microscopy (TEM) (HT7700; Hitachi, Japan) 185

with a JEOL JEM 12000 EX (equipped with a field emission gun) at 100 kV. The 186

cells suspension of M. comscasis WC007T was washed with Milli-Q water and 187

centrifuged at 5,000 g for 5 min. Subsequently, the sample was taken by immersing 188

copper grids coated with a carbon film for 20 min in the bacterial suspensions and 189

washed for 10 min in distilled water and dried for 20 min at room temperature [38]. 190

Strain WC007T was Gram-stain-negative and showed a curved rod-shaped, 191

1.0-3.5×0.5-0.8 µm in size, which had no flagellum as indicated by TEM (Fig. 2). 192

For phenotypic characteristics comparison, the temperature, pH and NaCl 193

concentration ranges for the growth of strain WC007T were determined in duplicate 194


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experiments using the ORG medium as described above. Water baths and 195

thermostatic incubators were used to incubate bacterial cultures from 4 to 80 °C. The 196

pH of the culture medium was adjusted by 6 M HCl for low pH and 10% NaHCO3 197

(w/v) for high pH. The pH range for growth was tested from pH 3.0 to pH 11.0 (initial 198

pH at 30 °C) with increments of 0.5 pH units. Salt tolerance was determined by 199

directly weighing NaCl (0-100 g L−1) into the Hungate tubes before packaging the 200

autotrophic medium. Catalase activity was determined by observation of bubble 201

production after the application of 3% (v/v) hydrogen peroxide solution. Oxidase 202

activity was evaluated by the oxidation of 1% (w/v) tetramethyl p-phenylenediamine. 203

Substrate utilization was tested at atmospheric pressure in duplicate, using Hungate 204

tubes containing basal medium contained (L−1): 1.0 g NaHCO3, 1.0 g NH4Cl, 1.0 g 205

CH3COONa, 0.2 g MgSO4.7H2O, 0.5 g KH2PO4, 0.7 g cysteine hydrochloride, 1 mL 206

0.1% (w/v) resazurin. Single substrate (including cellulose, pectin, xylan, glucose, 207

acetate, maltose, butyrate, fructose, glycine, ethanol, formate, lactate, sucrose, sorbitol, 208

D-mannose) was added from sterile filtered stock solutions to the final concentration 209

at 20 mM, respectively. Cell culture without adding any other substrates was used as a 210

control. The pH was adjusted to 7.0 with NaOH. The cultures were incubated at 30 °C 211

for 7 days and then determined by spectrophotometry at 600 nm. For each substrate 212

was repeated three times. All tested substrates were listed in the species description. 213

The detailed physiological characteristics that differentiate the Maribellus luteus 214

XSD2T were listed as Table 1. Strain WC007T required NaCl for growth and growth 215

was observed at 0-5.0% NaCl (optimum: 1.0% NaCl). And growth occurred at 216

28-37 °C (optimum, 30 °C) and at pH 6.0-8.0 (optimum, pH 7.0). In addition, the 217

positive activities of acetate, maltose, fructose, ethanol, formate, lactate, sorbitol and 218

D-mannose in strain WC007T distinguished those in strain XSD2T. The differential 219

phenotypic characteristics between strain WC007T and the closely related type strain 220

XSD2T are shown in the Table 1. Overall, the phenotypic characterization supports 221

that strain WC007T represents a novel species of the genus Maribellus. 222


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For chemotaxonomic analysis, cells of WC007T were cultured and collected under 223

the same conditions unless stated otherwise, with the closely related type strains were 224

grown on the ORG solid medium for 7 d at 30 °C under the same condition. Cells of 225

WC007T and the closely related type strain were harvested from cultures at the 226

mid-exponential phase of growth and freeze-dried. Cellular fatty acids were extracted 227

and determined from dried cells by using GC (model 7890A, Agilent, USA) 228

according to the protocol of the Sherlock Microbial Identification System [39]. Polar 229

lipids were extracted and determined as described by Tindall et al [40]. 230

The predominant respiratory quinone of strain WC007T was MK-7, which is the 231

most frequently identified respiratory quinone in bacteria belonging to the family 232

Prolixibacteraceae [4]. The major cellular fatty acids (>10.0 %) in strain WC007T 233

were iso-C15:0, C16:0, summed feature 3 and summed feature 8. The amount of 234

iso-C15:0 in strain WC007T (14.07%) was higher than that found in strain XSD2T 235

(1.45%), while the amount of C18:0 and C18:1ω9c in strain WC007T (2.39%, 7.62%) 236

was lower than that found in strain XSD2T (17.33%, 16.57%), respectively. The major 237

polar lipids in strain WC007T were phosphatidylethanolamine, one unidentified 238

phospholipid, one unidentified aminolipid and one unidentified lipid (Fig. S2). Strain 239

WC007T was apparently different from the type strain XSD2T by the presence of one 240

unidentified phospholipid. 241

In summary, phylogenetic analysis of strain WC007T based on 16S rRNA gene 242

sequence similarities confirmed the distinctness of strain WC007T from the closely 243

related strain XSD2T. Moreover, based on a polyphasic taxonomic approach and 244

several phenotypic characteristics, strain WC007T distinguished from Maribellus 245

luteus XSD2T, the only recognized species of the genus Maribellus. Therefore, we 246

propose that strain WC007T was classified as the type strain of a novel species in the 247

genus Maribellus, for which the name Maribellus comscasis sp. nov. is proposed. 248

Description of Maribellus comscasis sp. nov. 249


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Maribellus comscasis (com.sca'sis. L. gen. pl. n. comscasis from the Center for Ocean 250

Mega-Science, Chinese Academy of Sciences). 251

Cells are Gram-stain-negative curve-shaped, 2.0-6.0 µm in length and 0.5-0.8 µm 252

in width. Facultative anaerobic and oxidase-positive. The temperature range for 253

growth is 28-37 °C with an optimum at 30 °C. Growing at pH values of 6.0-8.0 254

(optimum, pH 7.0). Growth occurs at NaCl concentrations between 0.0-5.0% with 255

optimum growth at 1.0% NaCl. By analyzing the hydrolysis of polysaccharides, the 256

growth is promoted significantly by cellulose, pectin and xylan. From the sole carbon 257

source utilization test, growth is stimulated by acetate, maltose, fructose, lactate, 258

sorbitol and D-mannose. Weak growth occurs with ethanol and formate. The major 259

polar lipids are phosphatidylethanolamine, one unidentified phospholipid, one 260

unidentified aminolipid, one unidentified lipid. Containing significant proportions 261

(>10%) of the cellular fatty acids iso-C15:0, C16:0, summed feature 3 (containing 262

C16:1ω7c and/or C16:1ω6c) and summed feature 8 (containing C18:1ω7c and/or 263

C18:1ω6c). 264

The type strain, WC007T (=KCTC 25169T =MCCC 1K04777T), was isolated from 265

the sediment of deep-sea cold seep, P.R. China. The DNA G+C content of the type 266

strain is 38.38%. 267

268

Funding information 269

This work was funded by the National Key R and D Program of China (Grant No. 270

2018YFC0310800), China Ocean Mineral Resources R&D Association Grant (Grant 271

No. DY135-B2-14), Strategic Priority Research Program of the Chinese Academy of 272

Sciences (Grant No. XDA22050301), the Taishan Young Scholar Program of 273

Shandong Province (tsqn20161051), and Qingdao Innovation Leadership Program 274

(Grant No. 18-1-2-7-zhc) for Chaomin Sun. 275

Conflicts of interest 276

The authors have no conflict of interest. 277


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27. Koren S, Walenz BP, Berlin K, Miller JR, Bergman NH et al. Canu: scalable 358 and accurate long-read assembly via adaptive k-mer weighting and repeat separation. 359 Genome Res 2017;27(5):722-736. 360 28. Walker BJ, Abeel T, Shea T, Priest M, Abouelliel A et al. Pilon: An Integrated 361 Tool for Comprehensive Microbial Variant Detection and Genome Assembly 362 Improvement. Plos One 2014;9(11). 363 29. Richter M, Rossello-Mora R, Glockner FO, Peplies J. JSpeciesWS: a web 364 server for prokaryotic species circumscription based on pairwise genome comparison. 365 Bioinformatics 2016;32(6):929-931. 366 30. Richter M, Rossello-Mora R. Shifting the genomic gold standard for the 367 prokaryotic species definition. P Natl Acad Sci USA 2009;106(45):19126-19131. 368 31. Medlar AJ, Toronen P, Holm L. AAI-profiler: fast proteome-wide exploratory 369 analysis reveals taxonomic identity, misclassification and contamination. Nucleic 370 Acids Res 2018;46(W1):W479-W485. 371 32. Meier-Kolthoff JP, Auch AF, Klenk HP, Goker M. Genome sequence-based 372 species delimitation with confidence intervals and improved distance functions. Bmc 373 Bioinformatics 2013;14. 374 33. Hetharua B, Min DR, Liao H, Lin LA, Xu H et al. Litorivita pollutaquae gen. 375 nov., sp. nov., a marine bacterium in the family Rhodobacteraceae isolated from 376 surface seawater of Xiamen Port, China. Int J Syst Evol Micr 2018;68(12):3908-3913. 377 34. Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. MEGA6: Molecular 378 Evolutionary Genetics Analysis Version 6.0. Mol Biol Evol 2013;30(12):2725-2729. 379 35. Saitou N, Nei M. The Neighbor-Joining Method - a New Method for 380 Reconstructing Phylogenetic Trees. Mol Biol Evol 1987;4(4):406-425. 381 36. Felsenstein J. Evolutionary Trees from DNA-Sequences - a 382 Maximum-Likelihood Approach. J Mol Evol 1981;17(6):368-376. 383 37. Rzhetsky A, Nei M. A Simple Method for Estimating and Testing 384 Minimum-Evolution Trees. Mol Biol Evol 1992;9(5):945-967. 385 38. Han ZZ, Yan HX, Zhao H, Zhou SX, Han M et al. Bio-precipitation of Calcite 386 with Preferential Orientation Induced by Synechocystis sp. PCC6803. Geomicrobiol J 387 2014;31(10):884-899. 388 39. Athalye M, Noble WC, Minnikin DE. Analysis of Cellular Fatty-Acids by 389 Gas-Chromatography as a Tool in the Identification of Medically Important 390 Coryneform Bacteria. J Appl Bacteriol 1985;58(5):507-512. 391 40. Smibert R, Krieg N. Phenotypic characterization. Methods for general and 392 molecular bacteriology, p 607–654. Methods for general and molecular microbiology 393 ASM Press, Washington, DC 1994. 394

395

396

397

398


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Table 1. Differential physiological characteristics of the novel strain WC007T and its 399

closest related type strain Maribellus luteus XSD2T. Strains: 1, WC007T (all data from 400

this study); 2, Maribellus luteus XSD2T (all data from this study except DNA G+C 401

content and polar lipids). +, Positive result or growth; -, negative result or no growth. 402

Characteristic 1 2

Cell length (µm)

Temperature range

for growth (°C) Optimum

pH range for growth

Optimum

NaCl range for growth (%)

Optimum

Oxidase activity

Hydrolysis of:

Cellulose

Pectin

Xylan

Utilization as a sole carbon source:

Acetate

Maltose

Fructose

Ethanol

Formate

Lactate

Sorbitol

D-mannose

Major menaquinone(s)

Polar lipids

Major fatty acids (>10 %)

DNA G+C content (mol%)

Isolation source

2.0-6.0

28-37

30

6.0-8.0

7.0

0-5

1

+

+

+

+

+

+

+

-

-

+

+

+

MK-7

PE, PL, AL, L

iso-C15:0, C16:0, summed

feature 3, summed

feature 8

38.38

deep-sea sediments

2.0-9.0

20–40

28

6.0-8.5

7.0

1-5

2

-

-

-

-

-

-

-

+

+

-

-

-

MK-7

PE, AL, 3L


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C16:0, C18:1ω9c,

summed feature 8,

C18:0

44.1

surface seawater

*Summed features are groups of two or three fatty acids that could not be separated 403

by GLC using the MIDI system. Summed feature 8 contains C18:1ω7c and/or C18:1ω6c. 404

Summed feature 8 contains C16:1ω7c/C16:1ω6c and/or C16:1ω6c/C16:1ω7c. 405

Table 2. Percentages of fatty acids useful for distinguishing WC007T from its closest 406

relative Maribellus luteus XSD2T. Strains: 1, WC007T (all data from this study); 2, 407

Maribellus luteus XSD2T (all data from this study). 408

Fatty acid Percentage (w/v) of total fatty acids

1 2

Saturated:

C18:0

Branched:

2.39

17.33

iso-C15:0

C18:1ω9c

14.07

7.62

1.45

16.57

409

410

411

412

413

414

415

416

417

418


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419

420

421

422

423

424

425

426

427

428

Figure legends 429

Fig. 1. Neighbor-joining tree based on 16S rRNA gene sequences showing the 430

position of the novel strain WC007T among bacterial members of the family 431

Prolixibacteraceae and other closely related families. The filled circles indicate 432

branches of the tree that were also formed using the maximum-likelihood and 433

minimum-evolution methods. Numbers at branching points are bootstrap values 434

(expressed as percentages of 1000 replications). The access number of each 16S 435

rRNA is indicated after the strain’s name. The sequence of Agarivorans albus LMG 436

21761T is used as an outgroup. Bar, 0.05 substitutions per nucleotide position. 437

Fig. 2. Transmission electron microscopy (TEM) observation of a negatively stained 438

culture of strain WC007T. Bar is 2 μm. 439

440

441


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https://doi.org/10.1101/2021.01.06.425552

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