特殊生境微生物培养技术研究进展
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摘要
针对细菌耐药性日益增强,抗生素匮乏、特殊生境微生物药用资源开发不足等现状,对特殊生境微生物培养技术进行综述;总结制约特殊生境微生物生长的因素,概述设计开发新型的微生物培养基、改善微生物培养条件等方面研究进展,重点综述一些新颖独特的微生物培养技术和方法,包括群体培养与共培养、高通量培养技术、原位培养、细胞微囊包埋技术、分子生物学方法等;指出经典培养方法的优化使特殊生境微生物的可培养性显著改善,基于分子生物学方法的研究结果不仅丰富了微生物资源库,而且为今后特殊生境微生物分离与培养指出了明确方向。
In view of the increasingly severe drug resistance, the lack of antibiotics, and the inadequacy in the development of medicinal resources for special habitats, a review was made of microbial culture techniques in special habitats. The factors restricting the growth of microorganisms in special habitats were summarized, the research and development of the design and development of new microbial culture media were outlined, and the conditions of microbial culture were improved. Some of new and unique microbial culture techniques and methods were highlighted, including population culture and co-culture, high-throughput culture, in-situ culture, cell microencapsulation technology, and molecular biology methods. It was pointed out that the optimization of the classical culture method made the microorganism more culturable in special habitats. The results of the study based on the molecular biology method not only enriched the microbial resource pool, but also pointed out a clear direction for the isolation and culture of the special habitats in future.
In view of the increasingly severe drug resistance, the lack of antibiotics, and the inadequacy in the development of medicinal resources for special habitats, a review was made of microbial culture techniques in special habitats. The factors restricting the growth of microorganisms in special habitats were summarized, the research and development of the design and development of new microbial culture media were outlined, and the conditions of microbial culture were improved. Some of new and unique microbial culture techniques and methods were highlighted, including population culture and co-culture, high-throughput culture, in-situ culture, cell microencapsulation technology, and molecular biology methods. It was pointed out that the optimization of the classical culture method made the microorganism more culturable in special habitats. The results of the study based on the molecular biology method not only enriched the microbial resource pool, but also pointed out a clear direction for the isolation and culture of the special habitats in future.
引文
[1] 程东升. 资源微生物学[M]. 哈尔滨: 东北林业大学出版社, 1995.
[2] 车裕斌, 韩冰华. 特殊生境研究综述[J]. 咸宁师专学报(自然科学版),1997, 17(3): 74-75.
[3] 唐永红, 曹庸, 卢成瑛, 等. 特殊生境微生物及其活性代谢产物研究进展[J]. 微生物学通报, 2006, 33(4): 163-166.
[4] 岳秀娟, 余利岩, 张月琴. 自然界中处于VBNC状态微生物的研究进展[J]. 微生物学通报, 2004, 31(2): 108-111.
[5] VARTOUKIAN S R, PALMER R M, WADE W G. Strategies for culture of ‘unculturable’ bacteria[J]. FEMS Microbiology Letters, 2010, 309(1): 1-7.
[6] 付艳芳, 唐李斐, 万军庭, 等. 细菌的信息交流[J]. 氨基酸和生物资源, 2004, 26(4): 62-64.
[7] GUAN L L, ONUKI H, KAMINO K. Bacterial growth stimulation with exogenous siderophore and synthetic N-acyl homoserine lactone autoinducers under iron-limited and low-nutrient conditions[J]. Applied and Environmental Microbiology, 2000, 66(7): 2797-2803.
[8] BRUNS A, CYPIONKA H, OVERMANN J. Cyclic AMP and acyl homoserine lactones increase the cultivation efficiency of heterotrophic bacteria from the central Baltic Sea[J]. Applied and Environmental Microbiology, 2002, 68(8): 3978-3987.
[9] 王保军, 刘双江. 环境微生物培养新技术的研究进展[J]. 微生物学通报, 2013, 40(1): 6-17.
[10] 田甜, 李冬梅, 戴世鲲, 等. 海洋环境中难培养微生物的寡营养培养[J]. 微生物学通报, 2009, 36(7): 1031-1039.
[11] JANSSEN P H, YATES P S, GRINTON B E, et al. Improved culturability of soil bacteria and isolation in pure culture of novel members of the divisions acidobacteria, actinobacteria, proteobacteria, and verrucomicrobia[J]. Applied and Environmental Microbiology, 2002, 68(5): 2391-2396.
[12] FUQUA C, WINANS S C, GREENBERG E P. Census and consensus in bacterial ecosystems: the LuxR-LuxI family of quorum-sensing transcriptional regulators[J]. Annual Review of Microbiology, 1996, 50(1): 727-751.
[13] SANGWAN P, KOVAC S, DAVIS K E, et al. Detection and cultivation of soil verrucomicrobia[J]. Applied and Environmental Microbiology, 2005, 71(12): 8402-8410.
[14] HAMAKI T, SUZUKI M, FUDOU R, et al. Isolation of novel bacteria and actinomycetes using soil-extract agar medium[J]. Journal of Bioscience and Bioengineering, 2005, 99(5): 485-492.
[15] SANTINI J M, SLY L I, SCHNAGL R D, et al. A new chemolithoautotrophic arsenite-oxidizing bacterium isolated from a gold mine: phylogenetic, physiological, and preliminary biochemical studies[J]. Applied and Environmental Microbiology, 2000, 66(1): 92-97.
[16] SCHINK B, FRIEDRICH M. Phosphite oxidation by sulphate reduction[J]. Nature, 2000, 406(6791): 37.
[17] UPHOFF H U, FELSKE A, FEHR W, et al. The microbial diversity in picoplankton enrichment cultures: a molecular screening of marine isolates[J]. FEMS Microbiology Ecology, 2001, 35(3): 249-258.
[18] TYSON G W, BANFIELD J F. Cultivating the uncultivated: a community genomics perspective[J]. Trends in Microbiology, 2005, 13(9): 411-415.
[19] RETTEDAL E A, GUMPERT H, SOMMER M O. Cultivation-based multiplex phenotyping of human gut microbiota allows targeted recovery of previously uncultured bacteria[J]. Nature Communications, 2014, 5(5): 4714-4714.
[20] KAWANISHI T, SHIRAISHI T, OKANO Y, et al. New detection systems of bacteria using highly selective media designed by SMART: selective medium-design algorithm restricted by two constraints[J]. PLoS One, 2011, 6(1): e16512.
[21] TAMAKI H, SEKIGUCHI Y, HANADA S, et al. Comparative analysis of bacterial diversity in freshwater sediment of a shallow eutrophic lake by molecular and improved cultivation-based techniques[J]. Applied and Environmental Microbiology, 2005, 71(4): 2162-2169.
[22] LEADBETTER J R. Cultivation of recalcitrant microbes: cells are alive, well and revealing their secrets in the 21st century laboratory[J]. Current Opinion in Microbiology, 2003, 6(3): 274-281.
[23] SAIT M, HUGENHOLTZ P, JANSSEN P H. Cultivation of globally distributed soil bacteria from phylogenetic lineages previously only detected in cultivation-independent surveys[J]. Environmental Microbiology, 2002, 4(11): 654-666.
[24] DAVIS K E, JOSEPH S J, JANSSEN P H. Effects of growth medium, inoculum size, and incubation time on culturability and isolation of soil bacteria[J]. Applied and Environmental Microbiology, 2005, 71(2): 826-834.
[25] JANSSEN P H, ZENGLER K. New cultivation strategies for terrestrial microorganisms[M]//ZENGLER K. Accessing Uncultivated Microorganisms. Washington D C: ASM Press, 2008: 173-192.
[26] MITEVA V I, BRENCHLEY J E. Detection and isolation of ultrasmall microorganisms from a 120,000-year-old greenland glacier ice core[J]. Applied and Environmental Microbiology, 2005, 71(12): 7806-7818.
[27] DEDYSH S N. Cultivating uncultured bacteria from northern wetlands: knowledge gained and remaining gaps[J]. Frontiers in Microbiology, 2011, 2(184): 184.
[28] SAIT M, DAVIS K E R, JANSSEN P H. Effect of pH on isolation and distribution of members of subdivision 1 of the phylum Acidobacteria occurring in soil[J]. Applied and Environmental Microbiology, 2006, 72(3): 1852-1857.
[29] JANSSEN P H, SCHUHMANN A, M RSCHEL E, et al. Novel anaerobic ultramicrobacteria belonging to the Verrucomicrobiales lineage of bacterial descent isolated by dilution culture from anoxic rice paddy soil[J]. Applied and Environmental Microbiology, 1997, 63(4): 1382-1388.
[30] ANGEL R, MATTHIES D, CONRAD R. Activation of methanogenesis in arid biological soil crusts despite the presence of oxygen[J]. PLoS One, 2011, 6(5): e20453.
[31] KAEBERLEIN T, LEWIS K, EPSTEIN S S. Isolating “uncultivable” microorganisms in pure culture in a simulated natural environment[J]. Science, 2002, 296(5570): 1127-1129.
[32] PLUGGE C M, STAMS A J. Enrichment of thermophilic syntrophic anaerobic glutamate-degrading consortia using a dialysis membrane reactor[J]. Microbial Ecology, 2002, 43(3): 379-387.
[33] EGAN M, MOTHERWAY M O, KILCOYNE M, et al. Cross-feeding by Bifidobacterium breve UCC2003 during co-cultivation with Bifidobacterium bifidum PRL2010 in a mucin-based medium [J]. BMC Microbiol, 2014, 14(1): 1-14.
[34] DUBEY G P, BEN-YEHUDA S. Intercellular nanotubes mediate bacterial com munication[J]. Cell, 2011, 144(4): 590-600.
[35] BUTTON D K, SCHUT F, QUANG P, et al. Viability and isolation of marine bacteria by dilution culture: theory, procedures, and initial results[J]. Applied and Environmental Microbiology, 1993, 59(3): 881-891.
[36] HAHNKE R L, BENNKE C M, FUCHS B M, et al. Dilution cultivation of marine heterotrophic bacteria abundant after a spring phytoplankton bloom in the North Sea[J]. Environmental Microbiology, 2015, 17(10): 3515-3526.
[37] CONNON S A, GIOVANNONI S J. High-throughput methods for culturing microorganisms in very-low-nutrient media yield diverse new marine isolates[J]. Applied and Environmental Microbiology, 2002, 68(8): 3878-3885.
[38] COLIN Y, CAUMETTE P, GUYONEAUD R. Contribution of enrichments and resampling for sulfate reducing bacteria diversity assessment by high-throughput cultivation[J]. Journal of Microbiological Methods, 2015, 110: 92-97.
[39] RAPPé M S, CONNON S A, VERGIN K L, et al. Cultivation of the ubiquitous SAR11 marine bacterioplankton clade[J]. Nature, 2002, 418(6898): 630-633.
[40] de BRUYN J C, BOOGERD F C, BOS P, et al. Floating filters, a novel technique for isolation and enumeration of fastidious, acidophilic, iron-oxidizing, autotrophic bacteria[J]. Applied and Environmental Microbiology, 1990, 56(9): 2891-2894.
[41] GAVRISH E, BOLLMANN A, EPSTEIN S, et al. A trap for in situ cultivation of filamentous actinobacteria[J]. Journal of Microbiological Methods, 2008, 72(3): 257-262.
[42] FERRARI B C, WINSLEY T, GILLINGS M, et al. Cultivating previously uncultured soil bacteria using a soil substrate membrane system [J]. Nature Protocols, 2008, 3(8): 1261-1269.
[43] 姜明国, 甘光华, 杨立芳, 等. 广西红树林根际土壤放线菌的原位培养分离及其活性筛选[J]. 海洋学报, 2015, 37(2): 55-64.
[44] ZENGLER K, TOLEDO G, RAPPE M, et al. Cultivating the uncultured[J]. Proceedings of the National Academy of Sciences of the United States of America, 2002, 99(24): 15681-15686.
[45] ZENGLER K, BEHRENDT L. Protocols for high-throughput isolation and cultivation[M]//McGENITY T J, TIMMIS K N, FERNANDEZ B N. Hydrocarbon and Lipid Microbiology Protocols. Berlin: Springer, 2014: 27-35.
[46] BENDOV E, KRAMARSKYWINTER E, KUSHMARO A. An in situ method for cultivating microorganisms using a double encapsulation technique[J]. FEMS Microbiology Ecology, 2009, 68(3): 363-371.
[47] JIANG C Y, DONG L, ZHAO J K, et al. High-throughput single-cell cultivation on microfluidic streak plates[J]. Applied and Environmental Microbiology, 2016, 82(7): 2210-2218.
[48] NICHOLS D, CAHOON N, TRAKHTENBERG E M, et al. Use of ichip for high-throughput in situ cultivation of “uncultivable” microbial species[J]. Applied and Environmental Microbiology, 2010, 76(8): 2445-2450.
[49] JUNG D, SEO E Y, EPSTEIN S S, et al. Application of a new cultivation technology, I-tip, for studying microbial diversity in freshwater sponges of Lake Baikal, Russia[J]. Fems Microbiology Ecology, 2014, 90(2): 417-423.
[50] LIEBNER S, HARDER J, WAGNER D. Bacterial diversity and community structure in polygonal tundra soils from Samoylov Island, Lena Delta, Siberia[J]. International Microbiology the Official Journal of the Spanish Society for Microbiology, 2008, 11(3): 195-202.
[51] ALBERTSEN M, HUGENHOLTZ P, SKARSHEWSKI A, et al. Genome sequences of rare, uncultured bacteria obtained by diff-erential coverage binning of multiple metagenomes[J]. Nature Biotechnology, 2013, 31(6): 533-538.
[52] 董燕, 宋程程, 黄鹤. 单细胞测序技术研究进展[J]. 化学工业与工程, 2015, 32(1): 71-78.
[53] 文小霞, 徐邦牢, 王伟鑫, 等. 微流控芯片快速鉴定多重细菌[J]. 分析化学, 2014, 42(6): 791-798.
[54] 陈炯, 屠丽红, 陈艳新, 等. 微流体芯片技术检测食源性致病菌的应用研究[J]. 上海预防医学, 2014, 26(6): 330-334.
[55] 崔庆锋, 王黎明, 刘志恒, 等. 酸性土壤中嗜酸稀有放线菌的多样性研究[J]. 微生物学报, 2004, 44(5): 571-575.
[2] 车裕斌, 韩冰华. 特殊生境研究综述[J]. 咸宁师专学报(自然科学版),1997, 17(3): 74-75.
[3] 唐永红, 曹庸, 卢成瑛, 等. 特殊生境微生物及其活性代谢产物研究进展[J]. 微生物学通报, 2006, 33(4): 163-166.
[4] 岳秀娟, 余利岩, 张月琴. 自然界中处于VBNC状态微生物的研究进展[J]. 微生物学通报, 2004, 31(2): 108-111.
[5] VARTOUKIAN S R, PALMER R M, WADE W G. Strategies for culture of ‘unculturable’ bacteria[J]. FEMS Microbiology Letters, 2010, 309(1): 1-7.
[6] 付艳芳, 唐李斐, 万军庭, 等. 细菌的信息交流[J]. 氨基酸和生物资源, 2004, 26(4): 62-64.
[7] GUAN L L, ONUKI H, KAMINO K. Bacterial growth stimulation with exogenous siderophore and synthetic N-acyl homoserine lactone autoinducers under iron-limited and low-nutrient conditions[J]. Applied and Environmental Microbiology, 2000, 66(7): 2797-2803.
[8] BRUNS A, CYPIONKA H, OVERMANN J. Cyclic AMP and acyl homoserine lactones increase the cultivation efficiency of heterotrophic bacteria from the central Baltic Sea[J]. Applied and Environmental Microbiology, 2002, 68(8): 3978-3987.
[9] 王保军, 刘双江. 环境微生物培养新技术的研究进展[J]. 微生物学通报, 2013, 40(1): 6-17.
[10] 田甜, 李冬梅, 戴世鲲, 等. 海洋环境中难培养微生物的寡营养培养[J]. 微生物学通报, 2009, 36(7): 1031-1039.
[11] JANSSEN P H, YATES P S, GRINTON B E, et al. Improved culturability of soil bacteria and isolation in pure culture of novel members of the divisions acidobacteria, actinobacteria, proteobacteria, and verrucomicrobia[J]. Applied and Environmental Microbiology, 2002, 68(5): 2391-2396.
[12] FUQUA C, WINANS S C, GREENBERG E P. Census and consensus in bacterial ecosystems: the LuxR-LuxI family of quorum-sensing transcriptional regulators[J]. Annual Review of Microbiology, 1996, 50(1): 727-751.
[13] SANGWAN P, KOVAC S, DAVIS K E, et al. Detection and cultivation of soil verrucomicrobia[J]. Applied and Environmental Microbiology, 2005, 71(12): 8402-8410.
[14] HAMAKI T, SUZUKI M, FUDOU R, et al. Isolation of novel bacteria and actinomycetes using soil-extract agar medium[J]. Journal of Bioscience and Bioengineering, 2005, 99(5): 485-492.
[15] SANTINI J M, SLY L I, SCHNAGL R D, et al. A new chemolithoautotrophic arsenite-oxidizing bacterium isolated from a gold mine: phylogenetic, physiological, and preliminary biochemical studies[J]. Applied and Environmental Microbiology, 2000, 66(1): 92-97.
[16] SCHINK B, FRIEDRICH M. Phosphite oxidation by sulphate reduction[J]. Nature, 2000, 406(6791): 37.
[17] UPHOFF H U, FELSKE A, FEHR W, et al. The microbial diversity in picoplankton enrichment cultures: a molecular screening of marine isolates[J]. FEMS Microbiology Ecology, 2001, 35(3): 249-258.
[18] TYSON G W, BANFIELD J F. Cultivating the uncultivated: a community genomics perspective[J]. Trends in Microbiology, 2005, 13(9): 411-415.
[19] RETTEDAL E A, GUMPERT H, SOMMER M O. Cultivation-based multiplex phenotyping of human gut microbiota allows targeted recovery of previously uncultured bacteria[J]. Nature Communications, 2014, 5(5): 4714-4714.
[20] KAWANISHI T, SHIRAISHI T, OKANO Y, et al. New detection systems of bacteria using highly selective media designed by SMART: selective medium-design algorithm restricted by two constraints[J]. PLoS One, 2011, 6(1): e16512.
[21] TAMAKI H, SEKIGUCHI Y, HANADA S, et al. Comparative analysis of bacterial diversity in freshwater sediment of a shallow eutrophic lake by molecular and improved cultivation-based techniques[J]. Applied and Environmental Microbiology, 2005, 71(4): 2162-2169.
[22] LEADBETTER J R. Cultivation of recalcitrant microbes: cells are alive, well and revealing their secrets in the 21st century laboratory[J]. Current Opinion in Microbiology, 2003, 6(3): 274-281.
[23] SAIT M, HUGENHOLTZ P, JANSSEN P H. Cultivation of globally distributed soil bacteria from phylogenetic lineages previously only detected in cultivation-independent surveys[J]. Environmental Microbiology, 2002, 4(11): 654-666.
[24] DAVIS K E, JOSEPH S J, JANSSEN P H. Effects of growth medium, inoculum size, and incubation time on culturability and isolation of soil bacteria[J]. Applied and Environmental Microbiology, 2005, 71(2): 826-834.
[25] JANSSEN P H, ZENGLER K. New cultivation strategies for terrestrial microorganisms[M]//ZENGLER K. Accessing Uncultivated Microorganisms. Washington D C: ASM Press, 2008: 173-192.
[26] MITEVA V I, BRENCHLEY J E. Detection and isolation of ultrasmall microorganisms from a 120,000-year-old greenland glacier ice core[J]. Applied and Environmental Microbiology, 2005, 71(12): 7806-7818.
[27] DEDYSH S N. Cultivating uncultured bacteria from northern wetlands: knowledge gained and remaining gaps[J]. Frontiers in Microbiology, 2011, 2(184): 184.
[28] SAIT M, DAVIS K E R, JANSSEN P H. Effect of pH on isolation and distribution of members of subdivision 1 of the phylum Acidobacteria occurring in soil[J]. Applied and Environmental Microbiology, 2006, 72(3): 1852-1857.
[29] JANSSEN P H, SCHUHMANN A, M RSCHEL E, et al. Novel anaerobic ultramicrobacteria belonging to the Verrucomicrobiales lineage of bacterial descent isolated by dilution culture from anoxic rice paddy soil[J]. Applied and Environmental Microbiology, 1997, 63(4): 1382-1388.
[30] ANGEL R, MATTHIES D, CONRAD R. Activation of methanogenesis in arid biological soil crusts despite the presence of oxygen[J]. PLoS One, 2011, 6(5): e20453.
[31] KAEBERLEIN T, LEWIS K, EPSTEIN S S. Isolating “uncultivable” microorganisms in pure culture in a simulated natural environment[J]. Science, 2002, 296(5570): 1127-1129.
[32] PLUGGE C M, STAMS A J. Enrichment of thermophilic syntrophic anaerobic glutamate-degrading consortia using a dialysis membrane reactor[J]. Microbial Ecology, 2002, 43(3): 379-387.
[33] EGAN M, MOTHERWAY M O, KILCOYNE M, et al. Cross-feeding by Bifidobacterium breve UCC2003 during co-cultivation with Bifidobacterium bifidum PRL2010 in a mucin-based medium [J]. BMC Microbiol, 2014, 14(1): 1-14.
[34] DUBEY G P, BEN-YEHUDA S. Intercellular nanotubes mediate bacterial com munication[J]. Cell, 2011, 144(4): 590-600.
[35] BUTTON D K, SCHUT F, QUANG P, et al. Viability and isolation of marine bacteria by dilution culture: theory, procedures, and initial results[J]. Applied and Environmental Microbiology, 1993, 59(3): 881-891.
[36] HAHNKE R L, BENNKE C M, FUCHS B M, et al. Dilution cultivation of marine heterotrophic bacteria abundant after a spring phytoplankton bloom in the North Sea[J]. Environmental Microbiology, 2015, 17(10): 3515-3526.
[37] CONNON S A, GIOVANNONI S J. High-throughput methods for culturing microorganisms in very-low-nutrient media yield diverse new marine isolates[J]. Applied and Environmental Microbiology, 2002, 68(8): 3878-3885.
[38] COLIN Y, CAUMETTE P, GUYONEAUD R. Contribution of enrichments and resampling for sulfate reducing bacteria diversity assessment by high-throughput cultivation[J]. Journal of Microbiological Methods, 2015, 110: 92-97.
[39] RAPPé M S, CONNON S A, VERGIN K L, et al. Cultivation of the ubiquitous SAR11 marine bacterioplankton clade[J]. Nature, 2002, 418(6898): 630-633.
[40] de BRUYN J C, BOOGERD F C, BOS P, et al. Floating filters, a novel technique for isolation and enumeration of fastidious, acidophilic, iron-oxidizing, autotrophic bacteria[J]. Applied and Environmental Microbiology, 1990, 56(9): 2891-2894.
[41] GAVRISH E, BOLLMANN A, EPSTEIN S, et al. A trap for in situ cultivation of filamentous actinobacteria[J]. Journal of Microbiological Methods, 2008, 72(3): 257-262.
[42] FERRARI B C, WINSLEY T, GILLINGS M, et al. Cultivating previously uncultured soil bacteria using a soil substrate membrane system [J]. Nature Protocols, 2008, 3(8): 1261-1269.
[43] 姜明国, 甘光华, 杨立芳, 等. 广西红树林根际土壤放线菌的原位培养分离及其活性筛选[J]. 海洋学报, 2015, 37(2): 55-64.
[44] ZENGLER K, TOLEDO G, RAPPE M, et al. Cultivating the uncultured[J]. Proceedings of the National Academy of Sciences of the United States of America, 2002, 99(24): 15681-15686.
[45] ZENGLER K, BEHRENDT L. Protocols for high-throughput isolation and cultivation[M]//McGENITY T J, TIMMIS K N, FERNANDEZ B N. Hydrocarbon and Lipid Microbiology Protocols. Berlin: Springer, 2014: 27-35.
[46] BENDOV E, KRAMARSKYWINTER E, KUSHMARO A. An in situ method for cultivating microorganisms using a double encapsulation technique[J]. FEMS Microbiology Ecology, 2009, 68(3): 363-371.
[47] JIANG C Y, DONG L, ZHAO J K, et al. High-throughput single-cell cultivation on microfluidic streak plates[J]. Applied and Environmental Microbiology, 2016, 82(7): 2210-2218.
[48] NICHOLS D, CAHOON N, TRAKHTENBERG E M, et al. Use of ichip for high-throughput in situ cultivation of “uncultivable” microbial species[J]. Applied and Environmental Microbiology, 2010, 76(8): 2445-2450.
[49] JUNG D, SEO E Y, EPSTEIN S S, et al. Application of a new cultivation technology, I-tip, for studying microbial diversity in freshwater sponges of Lake Baikal, Russia[J]. Fems Microbiology Ecology, 2014, 90(2): 417-423.
[50] LIEBNER S, HARDER J, WAGNER D. Bacterial diversity and community structure in polygonal tundra soils from Samoylov Island, Lena Delta, Siberia[J]. International Microbiology the Official Journal of the Spanish Society for Microbiology, 2008, 11(3): 195-202.
[51] ALBERTSEN M, HUGENHOLTZ P, SKARSHEWSKI A, et al. Genome sequences of rare, uncultured bacteria obtained by diff-erential coverage binning of multiple metagenomes[J]. Nature Biotechnology, 2013, 31(6): 533-538.
[52] 董燕, 宋程程, 黄鹤. 单细胞测序技术研究进展[J]. 化学工业与工程, 2015, 32(1): 71-78.
[53] 文小霞, 徐邦牢, 王伟鑫, 等. 微流控芯片快速鉴定多重细菌[J]. 分析化学, 2014, 42(6): 791-798.
[54] 陈炯, 屠丽红, 陈艳新, 等. 微流体芯片技术检测食源性致病菌的应用研究[J]. 上海预防医学, 2014, 26(6): 330-334.
[55] 崔庆锋, 王黎明, 刘志恒, 等. 酸性土壤中嗜酸稀有放线菌的多样性研究[J]. 微生物学报, 2004, 44(5): 571-575.