In Situ Discrimination and Cultivation of Active Degraders in Soils by Genome-Directed Cultivation Assisted by SIP-Raman-Activated Cell Sorting
文献类型：期刊论文
| 作者 | Li, Jibing1,4,7,8; Zhang, Dayi2,3; Luo, Chunling1,4,7,8 ; Li, Bei5,6; Zhang, Gan4,7,8
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| 刊名 | ENVIRONMENTAL SCIENCE & TECHNOLOGY
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| 出版日期 | 2023-10-12 |
| 卷号 | 57期号:44页码:17087-17098 |
| 关键词 | Raman-activated cell sorting stable-isotope probing genome-directed cultivation active toluene degrader genomic sequencing |
| ISSN号 | 0013-936X |
| DOI | 10.1021/acs.est.3c04247 |
| 英文摘要 | The identification and in situ cultivation of functional yet uncultivable microorganisms are important to confirm inferences regarding their ecological functions. Here, we developed a new method that couples Raman-activated cell sorting (RACS), stable-isotope probing (SIP), and genome-directed cultivation (GDC)-namely, RACS-SIP-GDC-to identify, sort, and cultivate the active toluene degraders from a complex microbial community in petroleum-contaminated soil. Using SIP, we successfully identified the active toluene degrader Pigmentiphaga, the single cells of which were subsequently sorted and isolated by RACS. We further successfully assembled the genome of Pigmentiphaga based on the metagenomic sequencing of C-13-DNA and genomic sequencing of sorted cells, which was confirmed by gyrB gene comparison and average nucleotide identity determination. Additionally, the genotypes and phenotypes of this degrader were directly linked at the single-cell level, and its complete toluene metabolic pathways in petroleum-contaminated soil were reconstructed. Based on its unique metabolic properties uncovered by genome sequencing, we modified the traditional cultivation medium with antibiotics, amino acids, carbon sources, and growth factors (e.g., vitamins and metals), achieving the successful cultivation of RACS-sorted active degrader Pigmentiphaga sp. Our results implied that RACS-SIP-GDC is a state-of-the-art approach for the precise identification, targeted isolation, and cultivation of functional microbes from complex communities in natural habitats. RACS-SIP-GDC can be used to explore specific and targeted organic-pollution-degrading microorganisms at the single-cell level and provide new insights into their biodegradation mechanisms. |
| WOS研究方向 | Engineering ; Environmental Sciences & Ecology |
| 语种 | 英语 |
| WOS记录号 | WOS:001101030300001 |
| 源URL | [http://ir.gig.ac.cn/handle/344008/79093]  |
| 专题 | 有机地球化学国家重点实验室 |
| 通讯作者 | Luo, Chunling |
| 作者单位 | 1.Univ Chinese Acad Sci, Beijing 100039, Peoples R China 2.Jilin Univ, Key Lab Groundwater Resources & Environm, Minist Educ, Changchun 130012, Peoples R China 3.Jilin Univ, Coll New Energy & Environm, Changchun 130021, Peoples R China 4.Chinese Acad Sci, Guangzhou Inst Geochem, Guangdong Hong Kong Macao Joint Lab Environm Pollu, Guangzhou 510640, Peoples R China 5.Chinese Acad Sci, State Key Lab Appl Opt, Changchun Inst Opt Fine Mech & Phys, Changchun 130033, Peoples R China 6.HOOKE Instruments Ltd, Changchun 130033, Peoples R China 7.CAS Ctr Excellence Deep Earth Sci, Guangzhou 510640, Peoples R China 8.Chinese Acad Sci, Guangzhou Inst Geochem, State Key Lab Organ Geochem, Guangzhou 510640, Peoples R China |
| 推荐引用方式 GB/T 7714 | Li, Jibing,Zhang, Dayi,Luo, Chunling,et al. In Situ Discrimination and Cultivation of Active Degraders in Soils by Genome-Directed Cultivation Assisted by SIP-Raman-Activated Cell Sorting[J]. ENVIRONMENTAL SCIENCE & TECHNOLOGY,2023,57(44):17087-17098. |
| APA | Li, Jibing,Zhang, Dayi,Luo, Chunling,Li, Bei,&Zhang, Gan.(2023).In Situ Discrimination and Cultivation of Active Degraders in Soils by Genome-Directed Cultivation Assisted by SIP-Raman-Activated Cell Sorting.ENVIRONMENTAL SCIENCE & TECHNOLOGY,57(44),17087-17098. |
| MLA | Li, Jibing,et al."In Situ Discrimination and Cultivation of Active Degraders in Soils by Genome-Directed Cultivation Assisted by SIP-Raman-Activated Cell Sorting".ENVIRONMENTAL SCIENCE & TECHNOLOGY 57.44(2023):17087-17098. |
入库方式： OAI收割
来源：广州地球化学研究所
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