新世纪微生物学者的一项重要任务——未培养微生物的分离培养

扼要介绍近年来有关分离培养未培养的微生物 (Unculturablemicroorganisms)、扩大生态环境微生物多样性的认识等方面取得的重要进展。首先 ,是采用非常规的 ,甚至认为有毒性的电子传递物质 ,获得了多种新生理型(physiotypes)的纯种微生物。另外 ,在多种生态域中分离出非同寻常的微生物 ,扩大了这些生境微生物的多样性的视野 ,如普遍存在于大洋的浮游细菌SAR11类群 ,其微小菌体呈半月形 ,在海水中细菌浓度可达 (10⁵～10⁶) mL ,基因组估计为 1.54Mb。更值得注意的是从极端高温环境分离的嗜热纳米古细菌属 (Nanoarchaeota) ,其基因组仅有500kb ,是已知原核生物的最小者。应重视的是在分离这些多样性微生物的过程中所发展的新型培养技术 ,如微滴胶囊化法和扩散小室法都是具有革命性的方法 ,既有通用性 ,又无需昂贵设备 ,对今后这一领域的发展将起重要推动作用

An Important Mission for Microbiologists in the New Century-Cultivation of the Unculturable Microorganisms

The present review summarizes the recent advances in cultivating the so-called recalcitrant microorganisms in various natural environments. Firstly, the use of new electron donors and acceptors reveals a list of unique and previously unrecognized physiotypes (Table 1), and the growth supporting reactions include oxidation of phosphite, arsenite, soil humic substances, benzene, Fe (II) and aryl-halides. The novel pure cultures obtained include: Desulfatogenum phosphitoxidans, lithochemotrophic arsenite oxidizing bacteria, Dechloromonas agitata, Dechalosoma suillum, and various Proteobacteria capable of decomposing soil humic substances anaerobically. Secondly, the adaption of nutrient-poor media (down to 1/100 of the traditional concentration), including sea water, soil extract and extension of natural habitats (insect hind-gut, submarine hot vent) are extremely rewarding. For example, cultivation of the ubiquitous SAR11 marine bacterioplankton clade reveals a phylum, small in cell size and genome, but dominating in ocean surface, thus constituting a huge biomass on earth. In addition, the nanosized hyperthermophilic Archaeon from a submarine hot vent, represents an unknown phylum, and because of its symbiotic living with Ignicocous cells, it is named Nanoarchaeum equitans. Its genome size is only 500kb, the smallest for prokaryote. Considering its hyperthermic living environment and small genome, N. equitans may possibly still a primitive form of microbial life and affords an advantageous culture for the study of life origin and evolution on earth. Thirdly, several novel isolation techniques have been developed such as gel microdrop encapsulation and diffusion chamber, both with the advantages of high-throughout operation and simulating to certain extent of the natural environment, thus allowing growth synergy based on cross-feeding to occur. Finally, it should be noted that the cultivation of unculturable microorganisms mentioned in this review does not require expensive and sophisticated equipments and most of the techniques can be adapted in our laboratory at home.