Nonlabens ponticola sp. nov., isolated from seawater and reclassification of Nonlabens sediminis as a later heterotypic synonym of Nonlabens tegetincola

Abstract

A Gram-stain-negative, orange-pigmented and strictly aerobic bacterium, designated strain MJ115T, was isolated from seawater in Pohang, South Korea. Cells were non-motile rods and showed positive reactions for catalase and oxidase tests. Growth of strain MJ115T was observed at 4–35 °C (optimum, 30 °C), pH 6.0–7.0 (optimum, pH 6.5) and in the presence of 0–8.0 % (w/v) NaCl (optimum, 2.0%). Strain MJ115T contained iso-C15 : 0, anteiso-C15 : 0, anteiso-C17 : 1 ω9c, C17 : 0 2-OH, iso-C16 : 0 3-OH, iso-C17 : 0 3-OH and summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c) as major cellular fatty acids and menaquinone-6 as the major respiratory quinone. Phosphatidylethanolamine, two unidentified aminolipids and four unidentified lipids were detected as major polar lipids. The G+C content of the genomic DNA was 40.7 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain MJ115T formed a phyletic lineage with Nonlabens marinus S1-08T, Nonlabens agnitus JC2678T and Nonlabens antarcticus AKS 622T within the genus Nonlabens . Strain MJ115T was also most closely related to N. marinus S1-08T, N. agnitus JC2678T and N. antarcticus AKS 622T with 96.5, 96.4 and 96.0 % 16S rRNA sequence similarities, respectively. Here it is proposed that strain MJ115T represents a new species of the genus Nonlabens , for which the name Nonlabens ponticola sp. nov. is proposed. The type strain is MJ115T (=KCTC 72237T=NBRC 113963T). In addition, the comparison of the whole genome sequences and phenotypic features suggested that Nonlabens tegetincola and Nonlabens sediminis belong to the same species. Therefore, it is proposed that N. sediminis is reclassified as a later heterotypic synonym of N. tegetincola .

Alishewanella maricola sp. nov., isolated from seawater of the Yellow Sea

“…For the bacterial isolation, the collected seawater sample was serially diluted in artificial sea water (20 g NaCl, 2.9 g MgSO 4 , 4.53 g MgCl 2 ·6H 2 O, 0.64 g KCl, and 1.75 g CaCl 2 ·2H 2 O in per litre), and aliquots of each serial dilution were spread on marine agar (MA; BD, USA) and incubated at 30 °C for 5 days under aerobic conditions. Colonies grown on MA were randomly selected and their 16S rRNA genes were PCR-amplified using the universal primers F1 (5′-AGA GTT TGA TCM TGG CTC AG-3′) and R13 (5′-TAC GGY TAC CTT GTT ACG ACT T-3′) [9]. The PCR products were double-digested with Hae III and Hha I, and representative PCR amplicons indicating discrete fragment patterns were partially sequenced using the universal primer 340F (5′-CCT ACG GGA GGC AGC AG-3′), as described previously [9].…”

Rheinheimera maricola sp. nov., isolated from seawater of the Yellow Sea

“…The 16S rRNA gene of strain MA-13 T that was amplified by F1 and R13 primers was additionally sequenced using 518R (5′-ATTACCGCGGCTGCTGG-3′) and 805F (5′-GATTAGATACCCTGGTAGTC-3′) primers at Macrogen (Seoul, Republic of Korea) and the sequences obtained by the primers 340F, 518R and 805F were assembled to obtain an almost-complete 16S rRNA gene sequence (1476 nucleotides) as described previously [17]. The 16S rRNA gene sequence similarity values between strain MA-13 T and their closely related type strains were calculated using the EzBioCloud server (www.ezbiocloud.net/identify).…”

“…In brief, the seawater sample was serially diluted in artificial seawater (20 g NaCl, 2.9 g MgSO 4 , 4.53 g MgCl 2 ⋅6H 2 O, 0.64 g KCl and 1.75 g CaCl 2 ⋅2H 2 O per litre), spread on marine agar (MA; BD) and incubated at 30 °C for 5 days. The 16S rRNA genes in genomic DNA extracted from colonies grown on MA were amplified by PCR using the universal primers F1 (5′-AGAGTTTGATCMTGGCTCAG-3′) and R13 (5′-TACG GYTA CCTT GTTA CGACTT-3′) [17] and double-digested with HaeIII and HhaI [18]. Representative PCR products showing distinct fragment patterns were partially sequenced using the primer 340F (5′-CCTACGGGAGGCAGCAG-3′) [17].…”

“…A surface seawater sample was obtained at a distance of about 5 m from Chaeseokgang cliffs of the Yellow Sea (35° 37′ 30.4″ N 126° 27′ 58.0″ E) in the Republic of Korea. Strain MA-13 T was isolated from the seawater sample according to a previously described procedure, with some modifications [17]. In brief, the seawater sample was serially diluted in artificial seawater (20 g NaCl, 2.9 g MgSO 4 , 4.53 g MgCl 2 ⋅6H 2 O, 0.64 g KCl and 1.75 g CaCl 2 ⋅2H 2 O per litre), spread on marine agar (MA; BD) and incubated at 30 °C for 5 days.…”

“…The 16S rRNA genes in genomic DNA extracted from colonies grown on MA were amplified by PCR using the universal primers F1 (5′-AGAGTTTGATCMTGGCTCAG-3′) and R13 (5′-TACGGYTACCTTGTTACGACTT-3′) [17] and double-digested with Hae III and Hha I [18]. Representative PCR products showing distinct fragment patterns were partially sequenced using the primer 340F (5′-CCTACGGGAGGCAGCAG-3′) [17]. The resulting partial 16S rRNA gene sequences were compared with those of validated type strains using the Nucleotide Similarity Search program in the EzBioCloud server (www.ezbiocloud.net/identify/) [19] and a putatively novel strain in the genus Rheinheimera , designated as strain MA-13 T , was selected for further phenotypic and phylogenetic analysis.…”