Dubey Suresh Kumar, Tripathi Anil Kumar, Upadhyay Siddh Nath
Department of Botany, Banaras Hindu University, Varanasi 221 005, India.
Bioresour Technol. 2006 Nov;97(17):2217-24. doi: 10.1016/j.biortech.2005.06.008. Epub 2005 Sep 27.
Soil is a repository of diverse microorganisms, which has frequently been used to isolate and exploit microbes for industrial, environmental and agricultural applications. Knowledge about the structure and dynamics of bacterial communities in soil has been limited as only a small fraction of bacterial diversity is accessible to culture methods. Traditional enrichment techniques and the pure culture approach for microbiological studies have offered only a narrow portal for examining the soil microbial flora due to their limited selectivity. Therefore, the morphological and nutritional criteria used to describe bacterial community failed to provide a natural taxonomic order according to evolutionary relationship. Molecular methods under an emerging discipline of biology "molecular microbial ecology" are now helping in getting these constraints removed to some extent. Nucleic acid extraction from soil is the first crucial step in the application of most of the molecular techniques, which have largely been dominated by diverse variations of PCR. Due to its rapidity, sensitivity and specificity, PCR-based finger printing techniques have proved extremely useful in assessing the changes in microbial community structure. Such techniques can yield complex community profiles and can also provide useful phylogenetic information. Fluorescent in situ hybridization (FISH) can be used to evaluate the distribution and function of bacterial population in situ. DNA microarray techniques have also been developed and being frequently used for the evaluation of ecological role and phylogenetic affiliations of bacterial populations in the soil.
土壤是多种微生物的储存库,常被用于分离和利用微生物以用于工业、环境和农业应用。由于培养方法只能获取细菌多样性的一小部分,关于土壤中细菌群落结构和动态的知识一直有限。传统的富集技术和微生物学研究的纯培养方法由于其选择性有限,只为研究土壤微生物群落提供了一个狭窄的窗口。因此,用于描述细菌群落的形态学和营养标准未能根据进化关系提供自然的分类顺序。生物学新兴学科“分子微生物生态学”中的分子方法现在在一定程度上有助于消除这些限制。从土壤中提取核酸是大多数分子技术应用的关键第一步,这些技术在很大程度上由PCR的各种变体主导。基于PCR的指纹技术因其快速性、敏感性和特异性,在评估微生物群落结构变化方面已被证明极为有用。此类技术可以产生复杂的群落图谱,还能提供有用的系统发育信息。荧光原位杂交(FISH)可用于原位评估细菌群体的分布和功能。DNA微阵列技术也已得到发展,并经常用于评估土壤中细菌群体的生态作用和系统发育关系。