镉胁迫下稻田土壤微生物基因多样性的DGGE分子指纹分析

采用现代分子生物学技术,避开传统的分离培养过程,探讨重金属镉污染条件下稻田土壤微生物种群的基因多样性.经过直接从土壤中抽提总DNA,并对总DNA中16S rDNA及其中 V3可变区序列作PCR扩增、变性梯度凝胶电泳(DGGE)分析等,对镉胁迫下稻田土壤总DNA、微生物种类分布进行了初步的研究,发现不同浓度镉胁迫下稻田土壤间的菌种有明显差异,DGGE技术可以用于污染环境下微生物多样性研究.同时对DGGE分子指纹图谱的生物信息学分析作了初步尝试,为污染环境下土壤微生物多样性研究提供了新的实验方法与依据.

Diversity of Microbial Genes in Paddy Soil Stressed by Cadmium Using DGGE

Variations of diversity of microbial genes in submerged paddy soil stressed by heavy metal cadmium were studied using modern molecular biotechnology which includes directly extracting total DNA from paddy soil, amplifying 16S rDNA and their V3 variable region by PCR, the denaturing gradient gel electrophoresis (DGGE). Two methods for extraction and purification of microbial DNA were compared. Bacterial communities were quantified by analyzing the DGGE band patterns. The genetic clusters and correlative comparison of bacterial communities were analyzed based on the DGGE finger print. The results showed that there are some significant differences between bacterial communities in paddy soils treated with different concentrations of cadmium. The information about effect of cadium on microbial population based on molecular biological techniques are conformed with that from traditional methods,but that obtained about variations of microbial genes in paddy soil is much more than results based on the latter methods. It could provide a new way and foundation to research microbial gene diversity in contaminated environment.