Developing a microfluidic-based epicPCR reveals diversepotential hosts of the mcrA gene in marine cold seep  

作　　者：Wenli Shen Danrui Wang Jiangtao Li Yue Liu Yinzhao Wang Xingsheng Yang Xi Peng Bingliang Xie Lei Su Ziyan Wei Qing He Zhiyi Wang Kai Feng Wenbin Du Ye Deng 

机构地区：[1]CAS Key Laboratory of Environmental Biotechnology,Research Center for Eco-Environmental Sciences,Chinese Academy of Sciences,Beijing,China [2]Institute for MarineScience and Technology,Shandong University,Qingdao,China [3]Soil Ecology Lab,Nanjing Agricultural University,Nanjing,China [4]State Key Laboratory of Marine Geology,Tongji University,Shanghai,China [5]College of Environmental Science and Engineering,Liaoning Technical University,Fuxin,China [6]Microbiology Division,School of LifeSciences and Biotechnology,Shanghai Jiao Tong University,Shanghai,China [7]State Key Laboratory of Microbial Resources,Institute of Microbiology,Chinese Academy ofSciences,Beijing,China [8]Savaid Medical School,University of the Chinese Academy of Sciences,Beijin

出　　处：《mLife》2025年第1期70-82,共13页微生物(英文)

基　　金：supported by the NationalNatural Science Foundation of China(Nos.32001092,91851106,and 91951103).

摘　　要：Anaerobic methanotrophic(ANME)microbes play a crucial role in the bioprocess of anaerobic oxidation of methane(AOM).However,due to their unculturable status,their diversity is poorly understood.In this study,we established a microfluidics-based epicPCR(Emulsion,Paired Isolation,and Concatenation PCR)to fuse the 16S rRNA gene and mcrA gene to reveal thediversity of ANME microbes(mcrA gene hosts)in three sampling push-cores from the marine cold seep.A total of 372516Samplicon sequence variants(ASVs)of the mcrA gene hosts were detected,and classified into 78 genera across 23 phyla.Across all samples,the dominant phyla with high relative abundance(>10%)were the well-known Euryarchaeota,and somebacterial phyla such as Campylobacterota,Proteobacteria,and Chloroflexi;however,the specificity of these associations wasnot verified.In addition,the compositions of the mcrA gene hosts were significantly different in different layers,where thearchaeal hosts increased with the depths of sediments,indicating the carriers of AOM were divergent in depth.Furthermore,the consensus phylogenetic trees of the mcrA gene and the 16S rRNA gene showed congruence in archaea not in bacteria,suggesting the horizontal transfer of the mcrA gene may occur among host members.Finally,some bacterial metagenomeswere found to contain the mcrA gene as well as other genes that encode enzymes in the AOM pathway,which prospectivelypropose the existence of ANME bacteria.This study describes improvements for a potential method for studying the diversityof uncultured functional microbes and broadens our understanding of the diversity of ANMEs.

关 键 词：anaerobic oxidation of methane cold seep epicPCR mcrA gene 

分 类 号：Q938[生物学—微生物学]