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[目的]建立一种分析微生物肥料菌种稳定性的T-RFLP指纹图谱技术。[方法]采用T-RFLP指纹图谱技术对进口细菌型微生物肥料菌种的稳定性进行分析。[结果]样品中优势菌的T-RFs片段主要是87、246、247、330 bp,其中330 bp为主要片段,Shannon多样性指数和均一性指数测定结果表明,不同批次产品间差异性较小,微生物群落结构较稳定。[结论]建立了微生物肥料菌种稳定性分析的T-RFLP指纹图谱技术,为进出口微生物肥料产品提供了一种快速分析方法。 相似文献
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Intensive land use practices necessary for providing food and raw materials are known to have a deleterious effect on soil. However, the effects that such practices have on soil microbes are less well understood. To investigate the effects of land use intensification on soil microbial communities we used a combined T-RFLP and pyrosequencing approach to study bacteria, archaea and fungi in spring and autumn at five long term observatories (LTOs) in Europe; each with a particular land use type and contrasting levels of intensification (low and high). Generally, due to large gradients in soil variables, both molecular methods revealed that soil microbial communities were structured according to differences in soil conditions between the LTOs, more so than land use intensity. Moreover, variance partitioning analysis also showed that soil properties better explained the differences in microbial communities than land use intensity effects. Predictable responses in dominant bacterial, archaeal and fungal taxa to edaphic conditions (e.g. soil pH and resource availability) were apparent between the LTOs. Some effects of land use intensification at individual field sites were observed. However, these effects were manifest when land use change affected soil conditions. Uniquely, this study details the responses of different microbial groups to soil type and land use intensification, and their relative importance across a range of European field sites. These findings reinforce our understanding of drivers impacting soil microbial community structure at both field and larger geographic scales. 相似文献
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We investigated the abundance and genetic heterogeneity of bacterial nitrite reductase genes (nir) and soil structural properties in created and natural freshwater wetlands in the Virginia piedmont. Soil attributes included soil organic matter (SOM), total organic carbon (TOC), total nitrogen (TN), pH, gravimetric soil moisture (GSM), and bulk density (Db). A subset of soil attributes were analyzed across the sites, using euclidean cluster analysis, resulting in three soil condition (SC) groups of increasing wetland soil development (i.e., SC1 < SC2 < SC3; less to more developed or matured) as measured by accumulation of TOC, TN, the increase of GSM, and the decrease of Db. There were no difference found in the bacterial community diversity between the groups (p = 0.4). NirK gene copies detected ranged between 3.6 × 104 and 3.4 × 107 copies g−1 soil and were significantly higher in the most developed soil group, SC3, than in the least developed soil group, SC1. However, the gene copies were lowest in SC2 that had a significantly higher soil pH (~6.6) than the other two SC groups (~5.3). The same pattern was found in denitrifying enzyme activity (DEA) on a companion study where DEA was found negatively correlated with soil pH. Gene fragments were amplified and products were screened by terminal restriction fragment length polymorphism (T-RFLP) analysis. Among 146 different T-RFs identified, fourteen were dominant and together made up more than 65% of all detected fragments. While SC groups did not relate to whole nirK communities, most soil properties that identified SC groups did significantly correlate to dominant members of the community. 相似文献
4.
双价转基因抗虫棉花对土壤氨氧化细菌和氨氧化古菌群落结构及丰度的影响 总被引：2，自引：2，他引：0
采用末端片段多态性分析(T-RFLP)和实时定量PCR(Quantitative real-time PCR,qPCR)方法,研究了不同生长时期双价转基因抗虫棉花和亲本非转基因棉花(对照)土壤氨氧化细菌(Ammonia-oxidizing bacteria,AOB)和氨氧化古菌(Ammonia-oxidizing archaea,AOA)群落结构和丰度的变化,以探讨双价转基因抗虫棉花种植对土壤氨氧化微生物的影响。结果表明,双价转基因抗虫棉花土壤中AOB和AOA优势种群与对照相同,且随生长时期无显著变化,但在各自样品中所占比例不同。不同生长时期双价转基因抗虫棉花土壤AOB的Shannon指数和Evenness指数均与对照无显著差异,AOA的Shannon指数与对照也无显著差异,而苗期时Evenness指数显著低于对照(P0.05),其他3个生长时期差异均不显著。qPCR分析表明,除花铃期双价转基因抗虫棉花土壤AOB丰度高于对照外,其他3个生长时期均低于对照,而AOA丰度在4个生长时期均低于对照。吐絮期时,双价转基因抗虫棉花与对照土壤AOB和AOA丰度均差异显著(P0.05)。双价转基因抗虫棉花对土壤AOB和AOA群落结构组成无显著影响,但降低了AOB和AOA的数量,双价转基因抗虫棉花对土壤微生物的某些类群可能存在潜在影响。 相似文献
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To explore long-term impact of organic and inorganic fertilizers on microbial communities, we targeted both the total bacterial community and the autotrophic ammonia oxidizing bacteria (AOB) in soil from six treatments at an experimental field site established in 1956: cattle manure, sewage sludge, Ca(NO3)2, (NH4)2SO4, unfertilized and unfertilized without crops. All plots, except the bare fallows, were cropped with maize. Effects on activity were assessed by measuring the basal respiration and substrate induced respiration (SIR) rates, and the potential activity of the AOB. To determine the bacterial community composition, 16S rRNA genes were used to fingerprint total soil communities by terminal restriction fragment length polymorphism analysis and AOB communities by denaturing gradient gel electrophoresis. The fertilization regimes had clear effects on both activity and composition of the soil communities. Basal respiration and r, which was kinetically derived as the exponentially growing fraction of the SIR-response, correlated well with the soil organic C content (r=0.93 and 0.66, respectively). Soil pH ranged from 3.97 to 6.26 in the treatments and was found to be an important factor influencing all microbial activities. pH correlated negatively with the ratio between basal respiration and SIR (r=0.90), indicating a decreased efficiency of heterotrophic microorganisms to convert organic carbon into microbial biomass in the most acid soils with pH 3.97 and 4.68 ((NH4)2SO4 and sewage sludge fertilized plots, respectively). The lowest SIR and ammonia oxidation rates were also found in these treatments. In addition, these treatments exhibited individually different community fingerprints, showing that pH affected the composition of AOB and total bacterial communities. The manure fertilized plots harbored the most diverse AOB community and the pattern was linked to a high potential ammonia oxidation activity. Thus, the AOB community composition appeared to be more strongly linked to the activity than the total bacterial communities were, likely explained by physiological differences in the populations present. 相似文献
6.
水稻田是大气甲烷的重要排放源。尿素氮肥施用是提升水稻产量和品质的重要措施,但其对稻田土壤中产甲烷古菌的影响规律仍不清楚。通过模拟水稻生长季节可能的田间温度变化,本研究设置水稻土施加尿素(N,400 mg/kg干土)与未施加尿素两个处理,在15℃、25℃、37℃以及50℃下进行为期100天的厌氧培养,定期测定了培养过程中甲烷累积量以及土壤理化因子如p H、NH_4~+-N以及有机碳的变化,并运用基于16S r RNA基因的T-RFLP(末端限制性片段多态性分析)技术分析了产甲烷过程中古菌群落结构随时间的变化情况。结果表明:在中低温范围内(15~37℃),尿素对水稻土产甲烷有抑制作用,但在50℃高温下尿素对水稻土产甲烷量没有显著影响。尿素可能通过改变产甲烷古菌群落结构来影响甲烷的产生,在15~37℃范围内,尿素降低了水稻土产甲烷古菌群落的稳定性,增大了其在不同时间的差异性;而在50℃高温时,尿素对水稻土产甲烷古菌稳定性和差异性的影响不明显。不同温度下,尿素均降低了甲烷八叠球菌(Methanosarcinaceae)的丰度,且随着温度的变化,尿素对水稻土产甲烷机制的改变可能没有影响。 相似文献
7.
Both plants and microbes influence soil nutrient cycling. However, the links between plants, microbes and nutrient cycling are poorly understood. In this study, we investigated how plant identity and interspecific competition influence soil nitrogen cycling and attempted to link plant identity and interspecific competition to community structures of bacterial and archaeal ammonia oxidizers based on terminal restriction fragment length polymorphism analysis (T-RFLP) of bacterial and archaeal ammonia monooxygenase (amoA) genes. Faba bean and maize monocultures and a faba bean/maize mixture were planted with two nitrogen levels (0 and 100 mg N kg−1 soil as urea). Soil mineral nitrogen, ammonia oxidizer function (potential nitrification activity, PNA) and community structures were measured 28 and 54 days after plant emergence. Faba bean and maize substantially differed in their influences on mineral nitrogen concentrations and PNA in rhizosphere soils. Soil mineral nitrogen and PNA in the rhizosphere soils of the faba bean/maize mixture were closer to those of the maize monoculture than to those of the faba bean monoculture. T-RFLP with restriction enzymes BsaJI and Hpy8I distinguished variations in bacterial and archaeal ammonia oxidizers community structure, respectively, and detected both between-cluster and within-cluster variations in bacterial ammonia oxidizers. T-RFLP data showed that nitrogen addition favored part of a Nitrosospira cluster 3b sequence type and suppressed part of a cluster Nitrosospira 3a sequence type of bacterial ammonia oxidizers, while it had no influence on the archaeal ammonia oxidizer community structure. Although multivariate analysis showed that the function and community structure of bacterial ammonia oxidizers were significantly correlated, plant species and interspecific competition did not significantly change the community structure of bacterial and archaeal ammonia oxidizers. These results indicate that plant species and interspecific competition regulate soil nitrogen cycling via a mechanism of other than alteration in the community structure of ammonia oxidizers as investigated by DNA based methods. 相似文献
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Total and active soil fungal communities in a native eucalypt forest and first rotation Pinus elliotti plantation were investigated by direct extraction of DNA and RNA from soil. Terminal restriction fragment length polymorphism (T-RFLP) analysis of internal transcribed spacer (ITS) and 18S rRNA profiles indicated that total and active fungal communities differed significantly in both forest types. This was supported by DGGE profile analysis on an individual plot basis for both forest types and when groups in the canonical analysis were redefined to allow comparison between forest types. Analyses of both ITS and 18S T-RFLP profiles indicated that conversion from native eucalypt forest to P. elliottii plantation may significantly alter total and active soil fungal communities. ITS DGGE (DNA) and 18S (RNA) profiles also suggested differences in fungal communities in the two forest types. No significant separation of the fungal communities in the two forest types was observed, however, when ITS DGGE (RNA) profiles were compared. Overall, the data suggest that conversion from native eucalypt forest to P. elliottii plantation at the Beerburrum State Forest in subtropical Australia has significantly altered soil fungal communities. 相似文献
10.
Barbara Kraigher Janez Hacin Ivan Mahne Ines Mandic-Mulec 《Soil biology & biochemistry》2006,38(9):2762-2771
Fen peatlands are specific wetland ecosystems containing high soil organic carbon (SOC). There is a general lack of knowledge about the microbial communities that abound in these systems. We examined the microbial activity and community structure in two fen soils differing in SOC content sampled from the Ljubljana Marsh under different seasonal conditions. Substrate-induced respiration and dehydrogenase activity were used as indicators of total microbial activity. Both methods indicated higher microbial activities in the fen soil with the higher SOC content on all dates of sampling. To determine whether the differences in microbial activity were associated with differences in the microbial community structures, terminal restriction fragment length polymorphism (T-RFLP) of bacterial 16S rRNA genes was performed. Comparison of the T-RFLP profiles revealed very similar community structures in both fens and in the two seasonal extremes investigated. This suggested a stable community structure in the two fens, which is not affected by the SOC content or seasonal variation. In addition, a bacterial 16S ribosomal RNA gene based clone library was prepared from the fen soil with the higher SOC content. Out of 114 clones analysed, approximately 53% belonged to the Proteobacteria, 23% to the Acidobacteria, 21% to a variety of other taxa, and less than 3% were affiliated with the Firmicutes. 相似文献