title: Single-cell analysis reveals gene-expression heterogeneity in syntrophic dual-culture of Desulfovibrio vulgaris with Methanosarcina barkeri.
abstract: Microbial syntrophic metabolism has been well accepted as the heart of how methanogenic and other anaerobic microbial communities function. In this work, we applied a single-cell RT-qPCR approach to reveal gene-expression heterogeneity in a model syntrophic system of Desulfovibrio vulgaris and Methanosarcina barkeri, as compared with the D. vulgaris monoculture. Using the optimized primers and single-cell analytical protocol, we quantitatively determine gene-expression levels of 6 selected target genes in each of the 120 single cells of D. vulgaris isolated from its monoculture and dual-culture with M. barkeri. The results demonstrated very significant cell-to-cell gene-expression heterogeneity for the selected D. vulgaris genes in both the monoculture and the syntrophic dual-culture. Interestingly, no obvious increase in gene-expression heterogeneity for the selected genes was observed for the syntrophic dual-culture when compared with its monoculture, although the community structure and cell-cell interactions have become more complicated in the syntrophic dual-culture. In addition, the single-cell RT-qPCR analysis also provided further evidence that the gene cluster (DVU0148-DVU0150) may be involved syntrophic metabolism between D. vulgaris and M. barkeri. Finally, the study validated that single-cell RT-qPCR analysis could be a valuable tool in deciphering gene functions and metabolism in mixed-cultured microbial communities.
authors.affiliation: 1] Laboratory of Synthetic Microbiology, School of Chemical Engineering &Technology, Tianjin University, Tianjin 300072, P.R. China [2] Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin 300072, P.R. China [3] SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), 300072, P.R. China.
authors.forename: Zhenhua
authors.initials: Z
authors.lastname: Qi
authors.affiliation: 1] Laboratory of Synthetic Microbiology, School of Chemical Engineering &Technology, Tianjin University, Tianjin 300072, P.R. China [2] Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin 300072, P.R. China [3] SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), 300072, P.R. China.
authors.forename: Guangsheng
authors.initials: G
authors.lastname: Pei
authors.affiliation: 1] Laboratory of Synthetic Microbiology, School of Chemical Engineering &Technology, Tianjin University, Tianjin 300072, P.R. China [2] Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin 300072, P.R. China [3] SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), 300072, P.R. China.
authors.forename: Lei
authors.initials: L
authors.lastname: Chen
authors.affiliation: 1] Laboratory of Synthetic Microbiology, School of Chemical Engineering &Technology, Tianjin University, Tianjin 300072, P.R. China [2] Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin 300072, P.R. China [3] SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), 300072, P.R. China.
authors.forename: Weiwen
authors.initials: W
authors.lastname: Zhang
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism*; D002478:Cells, Cultured; D018920:Coculture Techniques; D016969:Desulfovibrio vulgaris / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D017021:Methanosarcina barkeri / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D059010:Single-Cell Analysis / Q000379:methods*
chemical_list: D001426:Bacterial Proteins; D015415:Biomarkers; D012333:RNA, Messenger