Methanoculleus taiwanensis sp nov., a methanogen isolated from deep marine sediment at the deformation front area near Taiwan

A mesophilic, hydrogenotrophic methanogen, strain CYW4(T), was isolated from deep-sea sediment obtained by the Ocean Researcher I cruiser, ORI-961, in 2011. The sediment was from the deformation front area offshore of south-western Taiwan. Here, seismic reflections indicated that methane hydrates were abundant. The methanogenic substrates utilized by strain CYW4(T) were formate and H-2/CO2, but not acetate, secondary alcohols, methylamines, methanol and ethanol. Cells of strain CYW4(T) were non-motile, irregular cocci and 0.6-1.5 mu m in diameter. The S-layer protein had an M-r of 112 000. The optimum growth conditions were at 37 degrees C, pH 8.1 and 0.08 M NaCl. Growth of the strain was stimulated by acetate. The G+C content of the chromosomal DNA of strain CYW4(T) was 61 mol%. Phylogenetic analysis revealed that strain CYW4(T) was most closely related to Methanoculleus marisnigri JR1(T) (96.82% 16S rRNA gene sequence similarity). Based on the morphological, phenotypic and phylogenetic characteristics presented here, it is evident that strain CYW4(T) represents a novel species of the genus Methanoculleus, and the name Methanoculleus taiwanensis sp. nov. is proposed. The type strain is CYW4(T) (=BCRC AR10043(T) =NBRC 110782(T)). The optical density of cultures of strain CYW4(T) dropped abruptly upon entering the stationary growth phase. During this time numerous particles of approximately 50 nm in diameter were observed on and around the cells. This suggests that strain CYW4(T) harbours a lytic virus that is induced in the stationary phase, which is of interest because only a few lytic viruses have been reported in methanogens.