Characterization of Gymnodinialimonas Ceratoperidinii gen. nov.,sp. nov., a New Bacterium Isolated From Rare Marine Dinoflagellate Ceratoperidinium Margalefii

DOI: https://doi.org/10.21203/rs.3.rs-1126159/v1

Abstract

A bacterial strain, designated J12C1-MA-4T, was isolated from a liquid culture of dinoflagellate Ceratoperidinium margalefii. The bacterium was Gram-stain-negative, aerobic, and rod-shaped. Oxidase and catalase were positive. Optimal growth was observed at 30°C, pH 7.0, in the presence of 1% (w/v) NaCl. Phylogenetic analyses based on the 16S rRNA gene and 92 core genes sets indicated that the strain J12C1-MA-4T belongs to the family Rhodobacteraceae in the class Alphaproteobacteria and represented a separate taxon separated from other genera. 16S rRNA gene sequence of strain J12C1-MA-4T showed high similarities to Loktanella ponticola KCTC 42133T (95.74%), Pseudooctadecabacter jejudonensis KCTC 32525T (95.52%) and Jannaschia helgolandensis KCTC 12191T (95.31%) in the Rhodobacteraceae family. The genome length of strain J12C1-MA-4T was 3621968 bp with a DNA G+C content of 64.48 mol%. The major cellular fatty acids of strain J12C1-MA-4T were summed feature 8 (comprising C18:1ω7c and/or C18:1ω6c) (>10%). Phosphatidylglycerol (PG), phosphatidylcholine(PC), phospholipids (PL), lipids 1 (L1) and aminolipid (AL) were shown to be the major polar lipids. The sole predominant isoprenoid quinone is Q-10. Based on phylogenetic, phenotypic, chemotaxonomic and genomic features, strain J12C1-MA-4T is considered to represent a new species in the new genus of the family Rhodobacteraceae for which the name Gymnodinialimonas ceratoperidinii gen. nov., sp. nov. is proposed. The type strain is J12C1-MA-4T (=KCTC 82770T = GDMCC 1.2729T).

Introduction

Dinoflagellates play a major role in the dynamics of the Earth's oceans and climate (Moustafa et al. 2010). Vitamins are an important growth factor for the majority of dinoflagellates (Tang et al. 2010). Marine Rhodobacteraceae are major vitamin suppliers for dinoflagellates (Sanudo-Wilhelmy et al. 2014). The family Rhodobacteraceae (Liang et al. 2021) belongs to the order Rhodobacterales within the class Alphaproteobacteria. Members are Gram-stain-negative, callus or rod-shaped and non-spore-forming bacteria. The major respiratory quinone is ubiquinone-10 (Q-10) and the major cellular fatty acid is unsaturated fatty acid C18 : 1 ω7c. And Rhodobacteraceae strains usually contain phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), and phosphatidylcholine (PC) as the main polar lipids. Members of the family Rhodobacteraceae are found in various saline environmental habitats, such as sea water, sea grass, marine sediment and hypersaline lake.

In the present study, we describe strain J12C1-MA-4T, which was isolated from rare dinoflagellate C. margalefii during a study on the relationship between bacteria and marine dinoflagellate. Polyphasic taxonomic analyses revealed that the strain J12C1-MA-4T should be proposed as a novel species of a new genus in the family Rhodobacteraceae, named as Gymnodinialimonas ceratoperidinii gen. nov.,sp. nov.

Materials And Methods

Isolation and culture conditions

The bacterial strain was isolated from a liquid culture of the dinoflagellate Ceratoperidinium margalefii (J12C1), which was collected from the East China Sea (31◦ 59′ 53′′ N, 127◦ 41′ 59′′ E) (Figure 1). The dinoflagellate was isolated from seawater and the pure dinoflagellate was cultured in F/2 medium for 30 days. Collect 100 microliters from the seed culture sample and spread on MA agar (MA, Marine agar 2216) and then cultured at 25°C. After 3 days, single colonies were transferred onto a new MA agar plate under the same conditions. A round, yellow, smooth, strain named J12C1-MA-4T was stored at -80°C in a suspension containing 10% (w/v) skimmed milk, and was selected for polyphasic sorting, and incubated as usual with MA at 25°C. Jannaschia helgolandensis KCTC 12191T; Pseudooctadecabacter jejudonensis KCTC 32525T; Loktanella acticola KCTC 52837T; Loktanella ponticola KCTC 42133T; Alterinioella_nitratireducens KCTC 72738T were obtained from KCTC as closely related strains in order to further investigate the characteristics of strain J12C1-MA-4T.

DNA extraction, PCR and 16S rRNA-based phylogenetic analysis

Bacteria grown was used as a template on MA agar plates for 3 days. The DNA of strain F J12C1-MA-4T was extracted with a spin columns filtration method (QIAamp DNA mini kit). The 16S rRNA gene of strain J12C1-MA-4T was amplified by polymerase chain reaction (PCR) using two bacterial specific universal primers 27F and 1492R and then the purified PCR product was sequenced by Biofact using primers 518F and 805R. Sequences were viewed and assembled using Geneious Prime (https://www.geneious.com). The almost complete 16S rRNA gene sequence (1402 bp) was submitted to the GeneBank (https://www.ncbi.nlm.nih.gov/nuccore/MZ457320). Subsequently, the obtained 16S rRNA gene sequence was compared to the type strain sequences available on EzBioCloud server (http://www.ezbiocloud.net) (Yoon et al. 2017a). Phylogenetic analyses using the neighbor-joining, maximum-likelihood and maximum-parsimony algorithms were performed as described by Jiang et al.(Jiang et al. 2021).

Genome sequencing and analysis

Whole-genome sequencing was performed based on the PacBio RSII (Pacific Biosciences Inc.) and the Illumina HiSeq 2500 sequencing platform (Macrogen, Korea). The sequence data were assembled using the RS HGAP (v3.0) De novo assembler. Genome annotation was performed using the NCBI Prokaryotic Genome Annotation Pipeline (PGAP) (Tatusova et al. 2016). Using the Genome-to-Genome Distance Calculator web server (http://ggdc.dsmz.de/distcalc2.php) and the ANI calculator (www.ezbiocloud.net/tools/ani) (Yoon et al. 2017b) to calculate Digital DNA-DNA hybridization (dDDH) values and average nucleotide identity (ANI) values (J.P et al. 2013). AAI values between of complete genome of J12C1-MA-4T and each reference genome were calculated using an online AAI calculation tool available at the webserver (http://enve-omics.ce.gatech.edu/) (Luo et al. 2014). The up-to-date bacterial core genome (UBCG), consisting of 92 core genes, was used to construct a genome-wide phylogenetic tree after collecting the whole genome sequences of all closely related strains in NCBI GenBank as described by Na et al. (Na et al. 2018).

Morphological, physiological and chemotaxonomic characterization

Its morphological features were examined by using a scanning electron microscope (JEOL Ltd, Tokyo, Japan), samples were prepared as described by Li et al. (Li et al. 2019). To explore the optimal medium for J12C1-MA-4T, a variety of media were selected here for testing, such as Marine agar 2216 (MA, Difco), Tryptic Soy agar (TSA, Difco), Luria-Bertani agar (LB, Difco), Potato Dextrose agar (PD, Difco), Nutrient agar (NA, beef extract 3.0 g/L, peptone 5.0 g/L, and agar 15.0 g/L). The temperature range for growth was determined in MA medium at 25°C for 3 days at different temperatures of 4, 10, 15, 20, 25, 30, 35, 37, 40, 45, 50 and 60°C. Growth at different pH values (pH 3.0–12.0 in 1.0 unit intervals) (adjusted with 1N HCl and 1N NaOH, 1.0 pH unit interval) was measured in modified MA broth with 10 mM acetate or 10 mM Tris-HCl buffer described by Jiang et al. (Jiang et al. 2020). NaCl tolerance was tested in DW containing MA broth (NaCl concentrations of 0-12%). The pH and NaCl tolerance were monitored by measuring the absorbance of OD600 using a DU 700 UV-Vis spectrophotometer (Beckman Coulter). Motility was assessed on soft agar (0.4%; w/v) test tubes. Gram stain test using Gram stain kit (Difco). The bubbles produced after the addition of 3% (v/v) hydrogen peroxide solution determined the activity of catalase, which was determined by the appearance of a purple color change in the colonies after the addition of an oxidase reagent (bioMérieux). Other biochemical characteristics were tested using API 20NE, API 50CH and API ZYM (bioMérieux) according to the manufacturer's instructions.

The appropriate liquid medium was prepared, and strain J12C1-MA-4T and closely related strains were cultured in a shaker incubator, shaken for 3 days, and then the bacteria were harvested by centrifugation at 6000 rpm, lyophilized, and polar lipids and quinones were extracted with 100 mg of lyophilized cells. For the analysis of chemosynthetic properties, methods including polar lipids (Minnikin et al. 1984) and quinones (Collins et al. 1980) were used. Polar lipid spots were separated by two-dimensional thin-layer chromatography and spray dyes including 0.2% ninhydrin (Sigma-Aldrich), γ-naphthol, molybdenum blue (Sigma-Aldrich), 4% phosphomolybdic acid and Dragendorff solution for the identification of amino-containing lipids, glycolipids, phospholipids, total lipids and quaternary nitrogen-containing lipids, respectively. Cellular fatty acid methyl esters (FAMEs) were obtained from cells by saponification, methylation and extraction according to the protocol of Sherlock Microbial Identification System (MIDI, USA). Cellular FAMEs were separated by gas chromatography (6890) and identified and quantified using Sherlock Microbial Identification System software (Sasser 2006).

Results And Discussion

Molecular characteristics

Comparative analysis of 16S rRNA (1402 bp) gene sequences revealed that strain J12C1-MA-4T shows the highest similarity to L. ponticola KCTC 42133T (95.74%), P. jejudonensis KCTC 32525T (95.52%), J. helgolandensis KCTC 12191T (95.31%), and A. nitratireducens KCTC 72738T (93.9%), L. acticola KCTC 52837T (93.3%).

Phylogenetic analyses using the neighbor-joining, maximum-likelihood and maximum-parsimony algorithms were performed as described by Jiang et al. (Jiang et al. 2021). NJ, ML, and ME algorithms obtained similar results (Figure 2). Based on 16S rRNA phylogenetic analysis, strain J12C1-MA-4T is located on a separate branch in the family Rhodobacteraceae, next to the genus Loktanella, Planktomarina, Thalassobius, Pseudooctadecabacter and Octadecabacter.

The whole-genome length of strain J12C1-MA-4T was 3621968 bp. Using the whole-genome calculated the GC-content was 64.48%. After annotation with PGAP (GenBank accession number: CP079194), there were 3428 protein-coding genes, 42 transfer RNA genes, 3 ncRNA and 6 ribosomal RNA genes, including 2 5S ribosomal RNA genes, 2 16S rRNA genes, 2 23S rRNA genes. The 3406 genes were functionally classified according to clusters of orthologous groups (COG) assignments, and 31.5% of the total assigned COGs were classified in the unknown group, followed by the amino acid transport and metabolism (9.6%), energy production and conversion (5.9%), transcription (5.3%). The digital DDH values, ANI value and AAI of strain J12C1-MA-4T to close related strain J. helgolandensis KCTC 12191T; P. jejudonensis KCTC 32525T; L. acticola KCTC 52837T; L. ponticola KCTC 42133T; A. nitratireducens KCTC 72738T revealed 18.8%, 20.9%, -,19.3%, 18.7%, and 71.59%, 70.69%, -, 69.58%, 73.54 and 60.55%, 61.28%,-, 60.18%, 64.99% are shown in Table 2 respectively. The dDDH values were much lower than the threshold for proposing novel bacterial species (dDDH: 70 %) (Chun et al. 2018).ANI values for J12C1-MA-4T are below the identified genus score threshold compared to closely related species (ANI: 92–94%) (Palmer et al. 2020). In addition, the AAI values between J12C1-MA-4T and other closely related genomes in the Rhodobacteraceae family ranged from 60-65%, which is below the defined genus delimitation threshold. (AAI: 80%) (Luo et al. 2014). The phylogenomic tree showed that strain J12C1-MA-4T is also located on a separate branch in the family Rhodobacteraceae. Nonetheless, strain J12C1-MA-4T had a far phylogenetic distance to all type strains within Rhodobacteraceae (Fig. 2), suggesting that strain J12C1-MA-4T was quite different Rhodobacteraceae from all other type strains. Therefore, the results of the phylogenetic analyses suggest that J12C1-MA-4T can be classified as a novel genus and species in the family of Rhodobacteraceae. Additionally, genes related to the metabolism, including complete pathway about carbohydrate (glycolysis, gluconeogenesis, Pyruvate oxidation, TCA cycle, first carbon oxidation, PRPP biosynthesis, D-galactonate degradation), lipid (fatty acid biosynthesis, acyl-CoA synthesis, Phosphatidylcholine (PC) biosynthesis), were encoded in the genome. Metabolism of cofactors and vitamins (Coenzyme A biosynthesis, Lipoic acid biosynthesis, Molybdenum cofactor biosynthesis, Cobalamin biosynthesis) were found to be encoded in the genome.

Morphological, physiological and chemotaxonomic characteristics

Colonies grown on MA medium were yellow-colored, circular, smooth, and 1 mm in diameter after 3 days of incubation. Strain J12C1-MA-4T can be grown in the temperature ranges 4-37°C (optimum 30°C), salt concentration range 0-8% (optimum 1%), pH range 5-8 (optimum 7), and this strain is Gram-negative, aerobic, non-motile, catalase-positive, and oxidase-positive. Cells were rod-shaped in shape, 0.2–0.5 µm width, and 1.0–1.8 µm length (Figure 4). Additional characteristics of strain J12C1-MA-4T with closely related strains are shown in Tables 1 and 3.

The polar lipid profile of strain J12C1-MA-4T was dominated by phosphatidylglycerol (PG), phosphatidylcholine(PC), phospholipids (PL), lipids 1 (L1) and aminolipid (AL) (Figure S1), J12C1-MA-4T and reference strain both have phosphatidylglycerol (PG) and phosphatidylcholine(PC). The predominant isoprenoid quinone of strain J12C1-MA-4T was Q-10, which was consistent with other members of the family Rhodobacteraceae. Strain J12C1-MA-4T contained Summed feature 8 (comprising C18:1ω7c and/or C18:1ω6c) (79.14%) as the major fatty acids (>10%) (Table 4). Regarding fatty acid composition, strain J12C1-MA-4T had only Summed feature 8 (including C18:1ω7c and/or C18:1ω6c) with more than 10%, and the percentage was as high as 79.14%, the most abundant fatty acids in the reference strains were all summed feature 8, but for its reference strains, more than one component over 10%, for example, J. helgolandensis KCTC 12191T had summed feature 8 (40.66%), C18:0 (12.73%) and C19:0 cyclo ω8c (27.90%) as major components (>10%), Pseudooctadecabacter jejudonensis KCTC 32525T had C18:1 ω7c 11-methyl (10.72%), summed feature 8 (71.37%) as major components (>10%), Loktanella acticola KCTC 52837T is mainly composed of, C18:1 ω7c 11-methyl (11.24%), summed feature 8 (69.85%). These results indicate that strain J12C1-MA-4T was affiliated with the family Rhodobacteraceae, suggesting that strain J12C1-MA-4T is distinguished from the closely related strains.

Taxonomic conclusion

In summary, the phylogenetic and chemotaxonomic analysis suggest that strain J12C1-MA-4T belongs to the family Rhodobacteraceae. However, 16S rRNA similarity (below 96.9%), ANI, AAI, fatty acid composition, dDDH values as well as some physiological characteristics (such as API 20NE, API 50CH, and API ZYM test), show that strain J12C1-MA-4T was different with the closely related strains. Thus, strain J12C1-MA-4T represents a new species in the new genus of the family Rhodobacteraceae, and the name of Gymnodinialimonas ceratoperidinii gen. nov., sp. nov. is proposed.

Description of Gymnodinialimonas gen. nov.

Gymnodinialimonas(Gym.no.di.ni.a.li.mo'nas. N.L. pl. neut. n. Gymnodiniales, unarmored dinoflagellates; L. fem. n. monas, unit, cell; N.L. fem. n. Gymnodinialimonas, a monad isolated from unarmored dinoflagellates)

Cells are Gram-strain-negative, aerobic, oxidase and catalase were positive. Predominant respiratory quinone is Q-10. The predominant fatty acids are summed feature 8 (comprising C18:1ω7c and/or C18:1ω6c). The principal polar lipids were phosphatidylglycerol (PG), phosphatidylcholine(PC), phospholipids (PL), lipids 1 (L1) and aminolipid (AL). The type species of Gymnodinialimonas is Gymnodinialimonas centrodinii.

Description of Gymnodinialimonas ceratoperidinii sp. nov.

Gymnodinialimonas ceratoperidinii (ce.ra.to.pe.ri.di'ni.i. N.L. gen. n. ceratoperidrii of the dinoflagellate Ceratoperidinium margalefii, the source of the isolation)

Colonies were circular, yellow-colored, smooth, and 1 mm in diameter when cultured on MA medium at 25°C for 3 days. Strain can grow at MA, the temperature ranged from at 4–37°C (optimum 30°C), at pH 5–8.0 (optimum pH 7.0), and in the presence of 0-8% NaCl (optimum 1%). Cells were aerobic, Gram-negative, rod-shaped (1.0–1.8 X 0.2–0.5 µm), non-motile, catalase-positive, and oxidase-positive. API 20NE tests indicated positive reactions for urease. API 50CH test showed the production of acid from esculin ferric citrate. In the API ZYM kit, show the positive result of acid phosphatase, valine arylamidase, cystine arylamidase, naphthol-as-bi-phosphohydrolase, alkaline phosphatase. The principal polar lipids of strain J12C1-MA-4T were phosphatidylglycerol (PG), phosphatidylcholine(PC), phospholipids (PL), lipids 1 (L1) and aminolipid (AL). Q-10 is the predominant isoprenoid quinone. The principal fatty acids of strain J12C1-MA-4T were summed feature 8 (comprising C18:1ω7c and/or C18:1ω6c).

The type strain is J12C1-MA-4T (= KCTC 82770T = GDMCC 1.2729T), which has been isolated from the dinoflagellate Ceratoperidinium margalefii.

The GenBank accession numbers for the 16S rRNA gene and the whole-genome sequence of strain J12C1-MA-4T are MZ457320 and CP079194, respectively.

Declarations

Data availability

The GenBank accession numbers of the 16S rRNA gene and the whole-genome sequences of the strain J12C1-MA-4T are MZ457320 and CP079194, respectively. This strain can be obtained from the Korea Collection for Type Cultures (KCTC 82770T) and the Guangdong Microbial Culture Collection Center (GDMCC 1.2729T).

Funding

This work was supported by grant from the Korea Research Institute of Bioscience and Biotechnology (KRIBB) Research Initiative Program and by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (No. 2021R1C1C1008377).

Conflict of interest

The authors declare that there are no conflicts of interest.

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Tables

Table 1. Comparison of the physiological and biochemical differences between strain J12C1-MA-4T and phylogenetically related type strains in the family Rhodobacteraceae based on 16S rRNA gene and genome sequences.

1, Gymnodinialimonas ceratoperidinii J12C1-MA-4T; 2, Jannaschia  helgolandensis KCTC 12191T; 3, Pseudooctadecabacter jejudonensis KCTC 32525T; 4Loktanella acticola KCTC 52837T; 5, Loktanella ponticola KCTC 42133T; 6, Alterinioella nitratireducens KCTC 72738T; ND, not determined.

Characteristics

1

2a

3b

4c

5d

6e

Isolation source

Ceratoperidinium margalefii

North Sea water

Zone where the ocean and a freshwater spring meet

Seawater

Seawater

Seawater

Colony morphology

Circular, yellow, smooth

small,

whitish, shiny

circular, slightly convex, smooth, glistening, reddish orange

circular, slightly convex, glistening, smooth, greyish yellow

circular, slightly convex, smooth, glistening, greyish-yellow

circular, convex, smooth, and nontransparent

Growth condition

 

 

 

 

 

 

temperature range (oC) (optimum)

4–37 (30)

15–30 (25–30)

15–35 (30)

4–30 (25)

4–30 (25)

10–37 (30)

Motile

Non-motile

Non-motile

Non-motile

Non-motile

Non-motile

Non-motile

Cell size (μm)

1.0–1.8 X

0.2–0.5

1.9–3.2 X 0.7–1.1

0.2–0.5 X 0.8–5.0

0.4–1.2 X >0.4

0.2–0.7 X 0.4–7.0

0.8–1.2 X 1.0–2.5

pH range (optimum)

5.0–9.0 (7.0)

7.0–8.0 (ND)

6.0–8.0 (7.0–8.0)

6.0–8.0(7.0–8.0)

5.5–8.0(7.0–8.0)

6.5–9.5 (8.0)

NaCl concentration range (%) (optimum)

0–8 (1)

1–7 (ND)

1–6 (2)

0–8.0 (2.0-3.0)

0.5–6.0 (2)

1.0–11 (3.0)

Data from Wagner et al. (Wagner-Dobler et al. 2003); b Data from Billerbeck et al. (Billerbeck et al. 2015) ; c Data from Park et al. (Park et al. 2017);  d Data from Jung et al. (Jung et al. 2014); e Data from Kong et al. (Kong et al. 2021).

Table 2. Comparison of the genome features differences between strain J12C1-MA-4T and phylogenetically related type strains in the family Rhodobacteraceae based on 16S rRNA gene and genome sequences.

1, Gymnodinialimonas ceratoperidinii J12C1-MA-4T; 2, Jannaschia  helgolandensis KCTC 12191T; 3, Pseudooctadecabacter jejudonensis KCTC 32525T; 4Loktanella acticola KCTC 52837T; 5, Loktanella ponticola KCTC 42133T; 6, Alterinioella_nitratireducens KCTC 72738T; ND, not determined.


1

2a

3b

4c

5d

6e

ANI

-

71.59%

70.69%

-

69.58%

73.54%

AAI

-

60.55%

61.28%

-

60.18%

64.99%

dDDH

-

18.8%

20.9%

-

19.3%

18.7%

G+C

64.48 mol%

-

-

57.3 mol%

55.9 mol%

65.4 mol%

Data from Wagner et al. (Wagner-Dobler et al. 2003); b Data from Billerbeck et al. (Billerbeck et al. 2015) ; c Data from Park et al. (Park et al. 2017);  d Data from Jung et al. (Jung et al. 2014); e Data from Kong et al. (Kong et al. 2021).

Table 3. Biochemical differences between strain J12C1-MA-4T phylogenetically related type starins in the family Rhodobacteraceae based on 16S rRNA gene and genome sequences.

1, Gymnodinialimonas ceratoperidinii J12C1-MA-4T; 2, Jannaschia   helgolandensis KCTC 12191T; 3, Pseudooctadecabacter jejudonensis KCTC 32525T; 4Loktanella acticola KCTC 52837T; 5, Loktanella ponticola KCTC 42133T; 6, Alterinioella_nitratireducens KCTC 72738T; All data were obtained from parallel experiments in this study under certain defined conditions. +, positive; -, negative; w, weak. 

Characteristics

1

2

3

4

5

6

API 20NE




 

 

 

Indoleproduction (TRyptoPhane)

-

+

-

-

-

-

Arginine DiHydrolase

-

+

-

-

+

-

UREase

+

+

+

+

-

-

Acid production (API 50CH)

 

 

 

 

 

 

Glycerol

-

-

-

+

-

+

L-arabinose

W

W

-

-

-

-

L-xylose

-

W

-

-

-

-

D-fructose

-

W

-

-

-

-

D-mannose

-

-

-

-

+

-

D-mannitol

-

-

-

+

-

-

Arbutin

-

-

-

+

-

-

Esculin ferric citrate

+

+

+

+

+

+

D-turanose

-

-

-

+

-

-

D-fucose

W

W

-

-

-

-

L-fucose

-

W

-

-

-

-

Potassium 5-KetoGluconate

W

W

-

-

-

-

ZYM                                                                                    

 

 

 

 

 

 

Alkaline phosphatase

-

­+

+

-

+

+

Esterase(C8)

+

+

+

-

-

+

Leucine arylamidase

+

+

+

+

-

+

Valine arylamidase

W

W

-

-

-

-

Cystine arylamidase

W

-

-

-

-

-

Acid phosphatase

+

+

+

-

-

-

Naphthol-AS-BI-phosphohydrolase

W

W

+

+

+

+

α-Galactosidase

-

W

-

-

-

-

β-Galactosidase

-

+

-

-

-

-

α-glucosidase

-

-

-

+

-

-

β-glucosidase

-

-

-

+

-

-

                                                                                                                             

Table 4. Cellular fatty acid compositions (>1%) of the strain J12C1-MA-4T and phylogenetically related type strains in the family Rhodobacteraceae based on 16S rRNA gene and genome sequences.

1, Gymnodinialimonas ceratoperidinii J12C1-MA-4T; 2, Jannaschia   helgolandensis KCTC 12191T 3, Pseudooctadecabacter jejudonensis KCTC 32525T; 4Loktanella acticola KCTC 52837T; 5, Loktanella ponticola KCTC 42133T; 6, Alterinioella_nitratireducens KCTC 72738T; All data are from the present study. Values are presented as the percentages of total fatty acids. ND, not detected. Major components (> 10%) are shown in bold.

Fatty acids

1

2

3

4

5

6

Saturated

 


 

 

 

 

C16:0

4.11

0.69

3.73

7.71

4.79

19.94

C18:0

1.2

12.73

3.86

1.69

0.36

1.29

Hydroxy

 

 

 

 

 

 

C10:0 3OH

1.98

2.95

1.77

1.63

0.99

2.14

C12:0 3OH

ND

ND

ND

ND

ND

4.31

C12:1 3OH

ND

ND

4.51

3.27

3.16

ND

C16:0 2OH

3.52

ND

ND

ND

ND

3.07

C18:1 2OH

3.69

ND

ND

ND

ND

ND

Unsaturated

 

 

 

 

 

 

C20:2 ω6,9c

ND

1.53

ND

ND

ND

0.88

C18:1 ω7c 11-methyl

3.83

7.05

10.72

11.24

2.42

1.24

C19:0 cyclo ω8c

ND

27.90

ND

0.33

ND

31.58

Summed feature*

 

 

 

 

 

 

2

0.17

1.58

ND

ND

ND

ND

3

1.05

0.38

1.17

1.53

13.56

1.47

7

ND

1.76

1.74

0.75

0.33

ND

8

79.14

40.66

71.37

69.85

73.61

32.17

*There are two or three fatty acids divided into a group which cannot be separated by GLC with the MIDI System, stipulated as summed features. Summed feature 2 contains C12:0 aldehyde, unknown 10.928; Summed feature 3 contains C16:1 ω7c and/or C16:1 ω6c, C16:1 ω6c and/or C16:1 ω7c; Summed feature 7 contains C19 : 1ω6cand/or C19 : 1ω7cand/or cyclo C19 : 0.  Summed feature 8 contains C18:1 ω7c and/or C18:1 ω6c.