Soil bacterial and fungal community successions under the stress of chlorpyrifos application and molecular characterization of chlorpyrifos-degrading isolates using ERIC-PCR
Soil bacterial and fungal community successions under the stress of chlorpyrifos application and molecular characterization of chlorpyrifos-degrading isolates using ERIC-PCR
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DOI：
10.1631/jzus.b1300175
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发表时间：
2014-04-01
影响因子：
5.1
通讯作者：
Yu, Yun-long
中科院分区：
文献类型：
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作者：
Chen, Lie-zhong;Li, Yan-li;Yu, Yun-long
毒死蜱是近年来广泛使用的一种杀虫剂，它在施于作物或与土壤混用时会对土壤产生不良影响。采用巢式聚合酶链反应（PCR）和变性梯度凝胶电泳（DGGE）相结合的方法，研究了毒死蜱5和20 mg/kg处理土壤中细菌和真菌群落的演替。此外，能够有效地分解毒死蜱的菌株进行了分子分型，使用肠细菌重复基因间共识-PCR（ERIC-PCR）。在实验条件下，毒死蜱在土壤中的降解符合一级动力学方程，毒死蜱在5和20 mg/kg剂量下的半衰期分别为8.25和8.29 d。DGGE指纹图谱和主成分分析（PCA）结果表明，毒死蜱处理后土壤真菌群落组成发生了明显变化，处理前后土壤真菌群落结构明显分离。而在细菌群落结构方面，毒死蜱处理土壤样品在前30 d内差异明显，到60 d后恢复到与对照相似的水平，且随着时间的推移，毒死蜱处理土壤样品与对照土壤样品在PCA上的距离逐渐缩短，最终聚为一类。总之，我们的研究结果表明，毒死蜱的应用可以影响真菌群落结构在一个快速和持久的方式，而只影响细菌群落在一个临时的方式。最后，筛选出9株典型的ERIC型毒死蜱降解菌。
Chlorpyrifos is a widely used insecticide in recent years, and it will produce adverse effects on soil when applied on crops or mixed with soil. In this study, nested polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE) were combined to explore the bacterial and fungal community successions in soil treated with 5 and 20 mg/kg of chlorpyrifos. Furthermore, isolates capable of efficiently decomposing chlorpyrifos were molecular-typed using enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR). Under the experimental conditions, degradation of chlorpyrifos in soil was interpreted with the first-order kinetics, and the half-lives of chlorpyrifos at 5 and 20 mg/kg doses were calculated to be 8.25 and 8.29 d, respectively. DGGE fingerprint and principal component analysis (PCA) indicated that the composition of the fungal community was obviously changed with the chlorpyrifos treatment, and that samples of chlorpyrifos treatment were significantly separated from those of the control from the beginning to the end. While for the bacterial community, chlorpyrifos-treated soil samples were apparently different in the first 30 d and recovered to a similar level of the control up until 60 d, and the distance in the PCA between the chlorpyrifos-treated samples and the control was getting shorter through time and was finally clustered into one group. Together, our results demonstrated that the application of chlorpyrifos could affect the fungal community structure in a quick and lasting way, while only affecting the bacterial community in a temporary way. Finally, nine typical ERIC types of chlorpyrifos-degrading isolates were screened.