Ibekwe A M, Papiernik S K, Gan J, Yates S R, Yang C H, Crowley D E
George E. Brown Jr. Salinity Laboratory, USDA Agricultural Research Service, Riverside, California 92507, USA.
Appl Environ Microbiol. 2001 Jul;67(7):3245-57. doi: 10.1128/AEM.67.7.3245-3257.2001.
Agricultural soils are typically fumigated to provide effective control of nematodes, soilborne pathogens, and weeds in preparation for planting of high-value cash crops. The ability of soil microbial communities to recover after treatment with fumigants was examined using culture-dependent (Biolog) and culture-independent (phospholipid fatty acid [PLFA] analysis and denaturing gradient gel electrophoresis [DGGE] of 16S ribosomal DNA [rDNA] fragments amplified directly from soil DNA) approaches. Changes in soil microbial community structure were examined in a microcosm experiment following the application of methyl bromide (MeBr), methyl isothiocyanate, 1,3-dichloropropene (1,3-D), and chloropicrin. Variations among Biolog fingerprints showed that the effect of MeBr on heterotrophic microbial activities was most severe in the first week and that thereafter the effects of MeBr and the other fumigants were expressed at much lower levels. The results of PLFA analysis demonstrated a community shift in all treatments to a community dominated by gram-positive bacterial biomass. Different 16S rDNA profiles from fumigated soils were quantified by analyzing the DGGE band patterns. The Shannon-Weaver index of diversity, H, was calculated for each fumigated soil sample. High diversity indices were maintained between the control soil and the fumigant-treated soils, except for MeBr (H decreased from 1.14 to 0.13). After 12 weeks of incubation, H increased to 0.73 in the MeBr-treated samples. Sequence analysis of clones generated from unique bands showed the presence of taxonomically unique clones that had emerged from the MeBr-treated samples and were dominated by clones closely related to Bacillus spp. and Heliothrix oregonensis. Variations in the data were much higher in the Biolog assay than in the PLFA and DGGE assays, suggesting a high sensitivity of PLFA analysis and DGGE in monitoring the effects of fumigants on soil community composition and structure. Our results indicate that MeBr has the greatest impact on soil microbial communities and that 1,3-D has the least impact.
为了有效防治线虫、土壤传播病原体和杂草,以便种植高价值经济作物,农业土壤通常要进行熏蒸处理。采用基于培养的方法(Biolog)和不依赖培养的方法(磷脂脂肪酸[PLFA]分析以及直接从土壤DNA扩增的16S核糖体DNA[rDNA]片段的变性梯度凝胶电泳[DGGE]),研究了熏蒸剂处理后土壤微生物群落的恢复能力。在一个微观实验中,研究了甲基溴(MeBr)、甲基异硫氰酸酯、1,3 - 二氯丙烯(1,3 - D)和氯化苦施用后土壤微生物群落结构的变化。Biolog指纹图谱的差异表明,MeBr对异养微生物活性的影响在第一周最为严重,此后MeBr和其他熏蒸剂的影响程度要低得多。PLFA分析结果表明,所有处理中微生物群落都向以革兰氏阳性细菌生物量为主的群落转变。通过分析DGGE条带模式,对熏蒸土壤中不同的16S rDNA图谱进行了定量分析。计算了每个熏蒸土壤样品的香农 - 韦弗多样性指数H。除了MeBr处理的土壤(H从1.14降至0.13)外,对照土壤和熏蒸剂处理土壤之间保持了较高的多样性指数。培养12周后,MeBr处理样品中的H增加到0.73。对独特条带产生的克隆进行测序分析表明,MeBr处理样品中出现了分类学上独特的克隆,这些克隆主要与芽孢杆菌属和俄勒冈嗜热放线菌密切相关。Biolog分析中数据的变化远高于PLFA和DGGE分析,这表明PLFA分析和DGGE在监测熏蒸剂对土壤群落组成和结构的影响方面具有较高的灵敏度。我们的结果表明,MeBr对土壤微生物群落的影响最大,而1,3 - D的影响最小。