The Institute of Low Temperature Science, Hokkaido University, Kita-19, Nishi-8, Kita-ku, 060-0819 Sapporo, Japan.
The Institute of Low Temperature Science, Hokkaido University, Kita-19, Nishi-8, Kita-ku, 060-0819 Sapporo, Japan.
Syst Appl Microbiol. 2022 May;45(3):126320. doi: 10.1016/j.syapm.2022.126320. Epub 2022 Mar 25.
Lake Harutori is a brackish meromictic lake with a steep physicochemical gradient in shallow water. Anoxic water below the chemocline has been characterized by high concentrations of sulfide (>10 mM) and methane (>1.5 mM). Previously, we reported that uncultured bacteria in the SEEP-SRB1 group were major sulfate reducers in the lake [21], but knowledge of sulfur oxidation and methane metabolism was scarce. In this current study, the Lake Harutori microbial community structure in the mixolimnion (at depths of 1.5 m and 3.0 m), upper chemocline (3.5 m), and monimolimnion (4.5 m) was further investigated by 16S rRNA gene amplicon sequencing and catalyzed reporter deposition fluorescence in situ hybridization (CARD-FISH). Reads of type I and II methanotrophs were retrieved mainly from 3.5 m and above. Methanotrophic bacteria detected by CARD-FISH accounted for 3.1% of DAPI-stained cells at 3.5 m. Detection frequencies of reads affiliated with the genera Sulfurimonas and Thiomicrorhabdus, which are known to comprise sulfur oxidizers, were relatively high at 3.5 m. Methanogenic archaeal reads were retrieved from the monimolimnion and they affiliated with the genus Methanosaeta. CARD-FISH counts indicated that the cells of Methanosaeta/Methanosarcina/Methanomicrobiales accounted for up to 0.8% of the DAPI-stained cells in the monimolimnion. On the other hand, many of the reads retrieved primarily from the monimolimnion were affiliated with phylogenetically novel uncultured groups.
哈鲁托里湖是一个微咸的分层湖,浅水区域的理化梯度很陡。在化变层以下的缺氧水中,硫化物(>10 mM)和甲烷(>1.5 mM)的浓度很高。之前,我们报道过,SEEP-SRB1 组中的未培养细菌是湖中主要的硫酸盐还原菌[21],但对硫氧化和甲烷代谢的了解很少。在本研究中,通过 16S rRNA 基因扩增子测序和催化报告物沉积荧光原位杂交(CARD-FISH),进一步研究了混合层(水深 1.5 m 和 3.0 m)、上化变层(3.5 m)和下化变层(4.5 m)中的哈鲁托里湖微生物群落结构。I 型和 II 型甲烷营养菌的reads 主要从 3.5 m 及以上回收。在 3.5 m 处,通过 CARD-FISH 检测到的甲烷营养菌占 DAPI 染色细胞的 3.1%。在 3.5 m 处,属于硫氧化菌的 Sulfurimonas 和 Thiomicrorhabdus 属的检测频率相对较高。甲烷生成古菌的 reads 从下化变层回收,它们属于 Methanosaeta 属。CARD-FISH 计数表明,Methanosaeta/Methanosarcina/Methanomicrobiales 细胞占下化变层 DAPI 染色细胞的 0.8%。另一方面,从下化变层主要回收的许多 reads 与系统发育新颖的未培养群有关。