摘要
微生物培养及显微技术作为鉴定微生物种群的手段有很大的局限性 ,因为环境中大多数微生物处于“存活但不能培养”的状态。因此 ,不依赖于微生物培养的生物化学以及分子生物学方法正被广泛地用于微生物生态学研究。主要介绍了荧光技术 ,基于 PCR的分析技术和
The cultivation of microbes and microscopy techniques as a means to identify microbial communities have severe limitations, since the majority of microbes in the environment are viable but nonculturable. For this reason, the methods of biochemistry and molecular biology that are independent of culturing are being widely used in the studies of microbial ecology. In this paper, fluorescent technique, PCR-based technique, PLFA technique and others to characterize diversity of microorganisms are reviewed. Fluorescence-based microscopy techniques are widely used in microbial ecology. Such techniques include fluorescent protein, fluorescence staining for total counts, viability counts and fluorescence in situ hybridizaion(FISH). The advantage of FISH is that it does not need cell lysis and easy to perform. But the result affected by the difference of the cell membrane's penetration ability. It is powerful tool for studying population dynamics, tracking microorganisms released into the environment but can't identify microorganisms has not been cultured. PCR-based techniques include PCR-RFLP, PCR-SSCP, PCR-DGGE. They have been the most useful in providing information about microbial communities. The greatest advantage of those methods is avoiding cultivation which solves the big obstacle “VNBC”. The technology has been well established and applied into microbial communities researches of soil, biofilms, seawater, wetlands and et al. The disadvantages caused by the storage, extraction efficiency, lysis efficiency and PCR efficiency have been solved in some extent but still need further study. PLFA and FAME are exclusively present in living cell and only in cell membranes. Since unique fatty acids are indications of specific groups, such signature molecules can be bacteria taxonomic discriminator especially FAME which has been accepted. Because fatty acids are rapidly metabolized following cell death, this method is very fit for community dynamics study. The shortcoming is that the method cannot be used to characterize microorganisms to species. Another is it need careful operation because any variation in those signatures would give rise to false community estimates created by artifacts in the methods. In some research areas, only one method cannot serve well. Synthetically use two or more methods provides favorable result and avoids intrinsical shortcomings. Summarily, the use of molecular techniques for microbial ecology research provided new understanding of microbial community. They give us a more complete picture of this area.
出处
《生态学报》
CAS
CSCD
北大核心
2003年第5期988-995,共8页
Acta Ecologica Sinica
基金
国家"十五"科技攻关重点项目 (2 0 0 1 BA90 3 B)~~
