基于PCR-DGGE的重金属污染土壤微生物种群指纹分析

采用变性梯度凝胶电泳（DGGE）分析方法,研究沈阳市细河沿岸重金属污染土壤中微生物结构多样性、种群丰富度及种群结构的动态变化规律。结果表明：直接从土壤中抽提总DNA,对16SrRNAV3可变区序列进行扩增,DGGE指纹图谱分析,微生物种群结构和变化规律明显,同时也表现为重金属污染程度越重,多样性指数越低;其中细河底泥重金属含量严重超标,多样性指数最低。经方差分析,重金属污染情况与Shannon-wiener多样性指数呈负相关,其中重金属污染对微生物的均匀度指数影响最显著,并且锌污染对其影响最大为-0.985。表明PCR-DGGE技术可以用于污染环境下微生物多样性研究,也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。

Microbial communities in heavy metal polluted soils by PCR-DGGE fingerprint

The diversities of the microbial structures,the abundances of the microbial communities and the variations of microbial population in the heavy metal polluted soil of Xihe region,Shenyang,were investigated using denaturing gradient gel electrophoresis（DGGE） method.Here,we report our results based on direct extracting total DNA from the polluted soil,amplifying 16Sr RNA V3 variable sequence and analyzing the DGGE finger-print.Our investigation shows that the more serious heavy metal polluted soil the lower the diversity indexes of the microbial population.Especially,the diversity indexes in the dredging sludge,where the heavy metal content reaches the lowest among the samples.According to the analysis of variance,we found that there is a negative correlation between the heavy metal polluted situation and the Shannon-wiener diversity index,and heavy metal has a noticeable effect on the microbial evenness index,for example,the largest evenness index value is-0.985 which is caused by the Zinc pollution.Our investigation indicates the PCR-DGGE technique can be used to study the biodiversity in contaminated environment.This technique also could provide a new way and foundation to study microbial gene diversity in contaminated environment.