Laboratory of Microbiology, Faculty of Fisheries Sciences, Hokkaido University, Hakodate, Japan.
Aquaculture Department, Faculty of Fisheries and Marine Sciences, Universitas Diponegoro, Semarang, Indonesia.
PLoS One. 2022 Aug 10;17(8):e0271174. doi: 10.1371/journal.pone.0271174. eCollection 2022.
A Gram-staining-negative, aerobic bacterium, designated strain PT3T was isolated from laboratory-reared larvae of the Japanese sea cucumber Apostichopus japonicus. Phylogenetic analysis based on the 16S rRNA gene nucleotide sequences revealed that PT3T was closely related to Amphritea ceti RA1T (= KCTC 42154T = NBRC 110551T) and Amphritea spongicola MEBiC05461T (= KCCM 42943T = JCM 16668T) both with 98.3% sequence similarity, however, average nucleotide identity (ANI) and in silico DNA-DNA hybridization (in silico DDH) values among these three strains were below 95% and 70%, respectively, confirming the novelty of PT3T. Furthermore, the average amino acid identity (AAI) values of PT3T against other Amphritea species were on the reported genus delineation boundary (64-67%). Multilocus sequence analysis using four protein-coding genes (recA, mreB, rpoA, and topA) further demonstrated that PT3T, Amphritea ceti and Amphritea spongicola formed a monophyletic clade clearly separate from other members of the genus Amphritea. Three strains (PT3T, A. ceti KCTC 42154T and A. spongicola JCM 16668T) also showed higher similarities in their core genomes compared to those of the other Amphritea spp. Based on the genome-based taxonomic approach, Aliamphritea gen. nov. was proposed together with the reclassification of the genus Amphritea and Aliamphritea ceti comb. nov. (type strain RA1T = KCTC 42154T = NBRC 110551T), Aliamphritea spongicola comb. nov. (type strain MEBiC05461T = KCCM 42943T = JCM 16668T), and Aliamphritea hakodatensis sp. nov. (type strain PT3T = JCM 34607T = KCTC 82591T) were suggested.
一株革兰氏阴性需氧细菌,被命名为 PT3T 菌株,从实验室饲养的日本刺参(Apostichopus japonicus)幼虫中分离得到。基于 16S rRNA 基因核苷酸序列的系统发育分析显示,PT3T 与 Amphritea ceti RA1T(=KCTC 42154T=NBRC 110551T)和 Amphritea spongicola MEBiC05461T(=KCCM 42943T=JCM 16668T)密切相关,它们的序列相似度均为 98.3%,然而,这三个菌株之间的平均核苷酸同一性(ANI)和体外 DNA-DNA 杂交(in silico DDH)值分别低于 95%和 70%,证实了 PT3T 的新颖性。此外,PT3T 与其他 Amphritea 种属的平均氨基酸同一性(AAI)值处于已报道的属划分界限(64-67%)。使用四个蛋白编码基因(recA、mreB、rpoA 和 topA)进行的多位点序列分析进一步表明,PT3T、Amphritea ceti 和 Amphritea spongicola 形成了一个单系分支,与 Amphritea 属的其他成员明显分离。与其他 Amphritea spp. 相比,这三个菌株(PT3T、A. ceti KCTC 42154T 和 A. spongicola JCM 16668T)的核心基因组也具有更高的相似性。基于基于基因组的分类方法,提出了 Aliamphritea 属新属,同时对 Amphritea 属进行了重新分类,将 Aliamphritea ceti 组合新种(模式株 RA1T=KCTC 42154T=NBRC 110551T)、Aliamphritea spongicola 组合新种(模式株 MEBiC05461T=KCCM 42943T=JCM 16668T)和 Aliamphritea hakodatensis 新种(模式株 PT3T=JCM 34607T=KCTC 82591T)。