应用PCR-RFLP及PCR-TGGE技术监测农田土壤微生物短期动态变化

在一块农田里于一个月内连续采集5次土样，提取土样微生物总DNA，应用PCR-RFLP及PCR-TGGE技术监测土壤微生物的动态变化。结果表明，细菌16S rDNA和真菌18S rDNA的PCR-RFLP图谱在5个采样时间点上基本一致；细菌的PCR-TGGE图谱平均有40条带，其相似性在90％以上，真菌的平均有20条带，其相似性在70％以上。说明这块农田土壤细菌区系短期内变化不大，而真菌区系存在较小幅度的动态变化，并且细菌多样性远远高于真菌多样性。比较两种分子生物学方法的结果表明，PCR-TGGE技术比PCR-RFLP技术更能精确地反映土壤微生物的动态变化，并且能同时快速地对多个样品进行有效区分。

Preliminary study on short-term dynamics of agricultural soil microbial community by PCR-RFLP and PCR-TGGE techniques

We studied the short-term dynamics of agricultural soil microbial community by PCR-RFLP and PCR-TGGE techniques. The results showed that the PCR-RFLP patterns of bacterial 16S rDNA and fungal 18S rDNA were similar among 5 soil samples which were continually sampled from a field in one month, the similarity of bacterial community PCR-TGGE patterns were up to 90%, with 40 bands averagely per sample, and that of fungal community were up to 70%, with 20 bands averagely per sample. It demonstrated that in this field soil bacterial community was relatively stable in short-term period, while fungal community had some extent changes, and bacterial diversity was far greater than that of fungi. It also manifested that PCR-TGGE is superior to PCR- RFLP for detail dynamics of soil microbial community, and it is a rapid method to discriminate multiple samples simultaneously.