Faculty of Agriculture and Life Science, Hirosaki University, 3 Bunkyo-cho, Hirosaki, Aomori 036-8561, Japan.
Int J Syst Evol Microbiol. 2014 Apr;64(Pt 4):1296-1303. doi: 10.1099/ijs.0.059378-0. Epub 2014 Jan 14.
An obligately anaerobic bacterial strain designated T-1-35(T) was isolated as a dominant cultivable cellulose-degrading bacterium from soil of a Japanese rice field as an anaerobic filter-paper degrader. Cells of strain T-1-35(T) stained Gram-positive and were non-spore-forming rods with rounded ends, 0.8-1.0×3.5-15.0 µm, and motile by means of two to four polar flagella. Cells of strain T-1-35(T) exhibited pleomorphism: in aged cultures (over 90 days of incubation), almost all cells were irregularly shaped. Although no spore formation was observed, cells tolerated high temperatures, up to 90 °C for 10 min. The temperature range for growth was 15-40 °C, with an optimum at 35 °C. The pH range for growth was 5.5-9.0, with an optimum at pH 8.0-8.5 (slightly alkaliphilic). Strain T-1-35(T) fermented some carbohydrates to produce ethanol and lactate as the major products. Major cellular fatty acids were iso-C16 : 0 and iso-C13 : 0 3-OH. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain T-1-35(T) belonged to Clostridium rRNA cluster III. The closest relative of strain T-1-35(T) was Bacteroides cellulosolvens WM2(T), with 16S rRNA gene sequence similarity of 93.4 %. Phenotypic, physiological and molecular genetic methods demonstrated that strain T-1-35(T) was distinct from its phylogenetic relatives (members of Clostridium rRNA cluster III) because it predominantly produced ethanol, iso-C13 : 0 3-OH was a major cellular fatty acid and it always exhibited pleomorphism. On the basis of the results of a polyphasic taxonomic study, strain T-1-35(T) is considered to represent a novel genus and species, Anaerobacterium chartisolvens gen. nov., sp. nov. The type strain of Anaerobacterium chartisolvens is T-1-35(T) ( = DSM 27016(T) = NBRC 109520(T)). In addition, from the results of our phylogenetic analysis and its phenotypic features, the species Bacteroides cellulosolvens Murray et al. 1984 is proposed to be reclassified in the new genus Pseudobacteroides as Pseudobacteroides cellulosolvens gen. nov., comb. nov., with the type strain WM2(T) ( = ATCC 35603(T) = DSM 2933(T) = NRCC 2944(T)).
一株被命名为 T-1-35(T)的严格厌氧菌从日本稻田的土壤中作为厌氧滤纸降解菌被分离出来,是一种优势可培养纤维素降解菌。T-1-35(T) 菌株的细胞革兰氏阳性,呈无芽孢的圆形杆状,0.8-1.0×3.5-15.0 µm,通过两个到四个极生鞭毛运动。T-1-35(T) 菌株的细胞表现出多形性:在老化培养物(超过 90 天的培养物)中,几乎所有细胞都是不规则形状的。虽然没有观察到孢子形成,但细胞能耐受高温,高达 90°C 持续 10 分钟。生长的温度范围为 15-40°C,最适温度为 35°C。生长的 pH 范围为 5.5-9.0,最适 pH 为 8.0-8.5(略偏碱性)。T-1-35(T) 菌株能发酵一些碳水化合物,产生乙醇和乳酸作为主要产物。主要的细胞脂肪酸是 iso-C16:0 和 iso-C13:0 3-OH。基于 16S rRNA 基因序列的系统发育分析表明,T-1-35(T) 菌株属于 Clostridium rRNA 簇 III。T-1-35(T) 菌株与 Bacteroides cellulosolvens WM2(T) 最为密切相关,16S rRNA 基因序列相似度为 93.4%。表型、生理和分子遗传学方法表明,T-1-35(T) 菌株与亲缘关系较远的菌株(Clostridium rRNA 簇 III 的成员)不同,因为它主要产生乙醇,iso-C13:0 3-OH 是主要的细胞脂肪酸,并且总是表现出多形性。基于多相分类学研究的结果,T-1-35(T) 菌株被认为代表了一个新的属和种,Anaerobacterium chartisolvens 属,种。Anaerobacterium chartisolvens 的模式菌株是 T-1-35(T)(=DSM 27016(T)=NBRC 109520(T))。此外,根据我们的系统发育分析结果及其表型特征,建议将 Murray 等人于 1984 年命名的物种 Bacteroides cellulosolvens 重新分类到新属 Pseudobacteroides 中,作为 Pseudobacteroides cellulosolvens 属,种。nov.,其模式菌株为 WM2(T)(=ATCC 35603(T)=DSM 2933(T)=NRCC 2944(T))。