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【目的】了解有机磷杀虫剂毒死蜱对土壤细菌群落结构的作用。【方法】联合微生物平板计数法、末端限制性片段多态性分析,选择PRIMER 5进行群落结构分析,以研究不同浓度的毒死蜱对辣椒根围可培养和不可培养细菌群落结构的影响。【结果】毒死蜱施入后前30 d,3个处理组的可培养细菌较对照组具有显著差异(P<0.05),但在第30天后,处理组可培养细菌数均能恢复到对照水平。采用PRIMER 5对T-RFLP数据进行多角度分析发现,HaeⅢ酶切片段中,对照组C3、处理组Y0和Z2的细菌群落结构较整体聚类较远。HhaⅠ酶切片段中,150μg/g的毒死蜱处理组(Z0)在第0天表现出最大的群落差异。ANOSIM表明,以不同浓度的毒死蜱分组,各组间细菌群落组成差异不显著(HaeⅢ:Global R=0.041,P=0.168;HhaⅠ:Global R=-0.04,P=0.842);以不同取样时间分组时,细菌群落组成差异显著(HaeⅢ:Global R=0.304,P=0.001;HhaⅠ:Global R=0.28,P=0.001)。经SIMPER分析所有TRFs可知,对群落丰度贡献最大的片段分别为TRF239、TRF240、TRF241。在线比对得到其代表菌群有芽孢杆菌属(Bacillus sp.)、梭菌属(Clostridium sp.)、葡萄球菌属(Staphylococcus sp.)、八叠球菌属(Sarcina sp.)、假单胞菌属(Pseudomonas sp.)等。【结论】高浓度的毒死蜱会对土壤细菌群落产生影响,抑制根围细菌生长,从而遏制植物的健康生长,因而有必要及时采取措施以减少大量重复使用毒死蜱所带来的危害。
【Objective】 To understand the effect of organophosphate insecticide chlorpyrifos on soil bacterial community structure. 【Method】 Combined microbial plate counting method and restriction fragment length polymorphism analysis of end fragments were carried out to select the PRIMER 5 community structure analysis to study the effects of different concentrations of chlorpyrifos on the culturable and uncultured bacterial community structure in pepper rhizosphere. 【Results】 The results showed that the viable bacteria of the three treatment groups had significant difference (P <0.05) compared with the control group before the first 30 days of treatment. However, after 30 days, the number of culturable bacteria in the treatment group recovered to the control level. Multi-angle analysis of T-RFLP data using PRIMER 5 revealed that bacterial community structure of control group C3 and treatment group Y0 and Z2 were far away from the overall clustering in HaeⅢ digestion fragments. In the Hha I digestion fragment, the 150 μg / g chlorpyrifos treated group (Z0) showed the largest community difference on the 0th day. ANOSIM showed that there was no significant difference in the composition of bacterial community between the groups with different concentrations of chlorpyrifos (HaeⅢ: Global R = 0.041, P = 0.168; HhaⅠ: Global R = -0.04, P = 0.842) (HaeⅢ: Global R = 0.304, P = 0.001; HhaⅠ: Global R = 0.28, P = 0.001). According to SIMPER analysis of all TRFs, the fragments that contributed the most to community abundance were TRF239, TRF240 and TRF241, respectively. On-line comparison revealed that the representative strains were Bacillus sp., Clostridium sp., Staphylococcus sp., Sarcina sp., Pseudomonas, Pseudomonas sp. 【Conclusion】 High concentrations of chlorpyrifos can affect the bacterial community in the soil, inhibit the growth of rhizosphere bacteria, and restrain the healthy growth of plants. Therefore, it is necessary to take prompt measures to reduce the harm caused by the heavy use of chlorpyrifos.