PLFA法和DGGE法分析堆肥细菌群落变化

同时采用PLFA（Phospholipid fatty acid）谱图分析法和DGGE（Denaturing gradient gel electrophoresis）法分析农业废物堆肥化过程中细菌群落的变化。结果表明：（1）单体PLFA数据显示样品之间的群落结构有明显差异,随着堆肥进程的进行,群落在不断演替,PLFA数据可以较为明确地表征微生物生物量的变化,但不能给出具体的物种变化信息;（2）DGGE分析显示堆肥过程中的样点和DGGE条带数据都存在3大主要类别,堆肥过程中细菌群落结构至少经历了3个阶段的演替,即嗜温细菌群落、高温细菌群落和腐熟期细菌群落。堆肥过程是微生物群落结构与堆肥温度相互制约的过程;（3）虽然两类方法显示的信息并不完全一致,但也因此说明了不同方法显示的信息并不是绝对的,采用各类方法组合研究堆肥微生物的信息变化是很有必要的。     

Land-use history has a stronger impact on soil microbial community composition than aboveground vegetation and soil properties

The response of soil microbial communities following changes in land-use is governed by multiple factors. The objectives of this study were to investigate (i) whether soil microbial communities track the changes in aboveground vegetation during succession; and (ii) whether microbial communities return to their native state over time. Two successional gradients with different vegetation were studied at the W. K. Kellogg Biological Station, Michigan. The first gradient comprised a conventionally tilled cropland (CT), mid-succession forest (SF) abandoned from cultivation prior to 1951, and native deciduous forest (DF). The second gradient comprised the CT cropland, early-succession grassland (ES) restored in 1989, and long-term mowed grassland (MG). With succession, the total microbial PLFAs and soil microbial biomass C consistently increased in both gradients. While bacterial rRNA gene diversity remained unchanged, the abundance and composition of many bacterial phyla changed significantly. Moreover, microbial communities in the relatively pristine DF and MG soils were very similar despite major differences in soil properties and vegetation. After >50 years of succession, and despite different vegetation, microbial communities in SF were more similar to those in mature DF than in CT. In contrast, even after 17 years of succession, microbial communities in ES were more similar to CT than endpoint MG despite very different vegetation between CT and ES. This result suggested a lasting impact of cultivation history on the soil microbial community. With conversion of deciduous to conifer forest (CF), there was a significant change in multiple soil properties that correlated with changes in microbial biomass, rRNA gene diversity and community composition. In conclusion, history of land-use was a stronger determinant of the composition of microbial communities than vegetation and soil properties. Further, microbial communities in disturbed soils apparently return to their native state with time.     

Temporal dynamics and variation with forest type of phospholipid fatty acids in litter and soil of temperate forests across regions

Microorganisms form the basis of soil food webs and represent key control points of carbon cycling and sequestration. Virtually all central European forests are managed and land-use regimes likely impact microbial abundance and community composition. Consequently, knowledge on how land-use intensity and abiotic variables, such as pH, C-to-N ratios, moisture regimes and concomitantly different stress levels, affect microbial communities is needed. We investigated phospholipid fatty acid (PLFA) profiles of leaf litter and soil from four forest types differing in foliage, age and management intensity, replicated in three regions across Germany. To account for temporal variation, samples were taken twice in the same season, but with an interval of three years. Total microbial biomass and microbial community composition differed between years, presumably due to between year variations in weather conditions. The litter layer was more prone to effects of drying, with a reduction of almost 30% of total PLFAs in the drier year. In soil effects of weather conditions depended on soil type and therefore differed between regions, with microorganisms in the sandy soils of the Schorfheide being more susceptible to water-stress, as evidenced by a ten-fold increase of the stress indicator cy/pre ratio in the drier year. Despite temporal variations in microbial biomass and community composition, the balance between the fungal and bacterial energy channel, as measured by fungal-to-bacterial ratios, remained rather constant in particular in soil. While total microbial biomass did not differ between forest types, microbial community composition differed significantly between beech and coniferous forests. Despite more acidic conditions, the fungal energy channel was less pronounced in leaf litter of coniferous forests than in broad-leaved forests, whereas the proportion of bacterial fatty acids was the highest in coniferous forests. Increasing management intensity presumably fosters the bacterial energy channel in the exposed litter layer. Supporting this assumption coniferous forests featured significantly higher values of the stress indicators cy/pre and SAT/MONO ratio. Bacterial community structure and biomass closely correlated with pH, with particular PLFAs dominating at high and low pH, respectively, indicating pH-specific microbial communities. In contrast, fungal abundance in leaf litter was correlated with C-to-N ratio. The results suggest that leaf litter and soil need to be considered separately when investigating changes in microbial community composition, since susceptibility of microorganisms to environmental stressors differs markedly between these layers. This, and repeated sampling events, may be particularly important when investigating subtle effects such as those related to climate change.     

铜污染土壤线虫多样性的PCR-DGGE分析

A wheat pot experiment was conducted under greenhouse conditions to assess the effect of copper contamination on soil nematode diversity by polymerase chain reaction-denaturing gradient gel electrophoresis （PCR-DGGE） method and morphological analysis. The soil was treated with CuSO4.5H2O at the following concentrations： 0, 50, 100, 200, 400, and 800 mg kg^-1 dry soil, and the soil samples were collected at wheat jointing and ripening stages. Nematode diversity index （H′） from morphological analysis showed no difference between the control and the treated samples in either of the sampling dates. At the wheat ripening stage, nematode diversity obtained by the PCR-DGGE method decreased noticeably in the Cu800 treatment in comparison with the control. With optimization of the method of nematode DNA extraction, PCR-DGGE could give more information on nematode genera, and the intensity of the bands could reflect the abundance of nematode genera in the assemblage. The PCR-DGGE method proved promising in distinguishing nematode diversity in heavy metal coritaminated soil.     

Investigation of Associations Between Rhizosphere Microorganisms and the Chemical Composition of Flue-Cured Tobacco Leaves Using Canonical Correlation Analysis

Canonical correlation analysis was applied to determine the relationship of rhizosphere microorganisms to common chemical components in flue-cured tobacco leaves. The results were as follows: (1) The chemical composition of the tobacco was related to the major microbial physiological groups in the rhizosphere soil. No close relationship was found with the populations of bacteria, fungi, and actinomycetes. (2) Greater contents of total nitrogen and potassium in tobacco leaves were associated with larger populations of aerobic nitrogen-fixing bacteria and with smaller populations of aerobic cellulose-decomposing bacteria. In addition, potassium content was also negatively correlated with nitrosobacteria counts but showed no close relationship with the potassium bacteria in the rhizosphere. The nicotine content was associated with greater populations of nitrosobacteria and nitrobacteria, and the content of water-soluble total sugar was positively correlated with the counts of nitrosobacteria.     

Inputs of nitrogen and organic matter govern the composition of fungal communities in soil disturbed by overwintering cattle

Overwintering cattle outdoors causes soil surface disturbance, substantial increases of soil N_tot, C_org, and P and a shift in pH to alkaline levels. Since fungi predominate in unfertilized soils with acidic pH and have filamentous hyphae, we hypothesized that changes caused by overwintering cattle outdoors (trampling, excreta returns, and changes in soil chemistry) will lead to suppressed species richness, lower biomass, and alter the structure of fungal communities. The research was conducted on an upland pasture used more than 10 years for cattle overwintering. Both culture-dependent and -independent methods were used for the determination of either fungal species composition (cultivation; DGGE) or biomass (numbers of CFU; concentration of fungal PLFA marker 18:2ω6,9). Soils under three different levels of cattle disturbance (S - severe, M - moderate, C - no disturbance/control) were investigated during three subsequent years. In addition, the DGGE analysis of soils was completed by comparison with analysis of fresh cattle excrements (Ex). The composition of fungal communities showed significantly higher richness and a substantial shift in species composition in cattle-disturbed soils (S, M) in comparison to the non-disturbed soil (C). The number of separated DGGE bands was significantly higher in S (30.67 ± 1.63; mean ± SD) and M (25.50 ± 1.64) soils than in the C soil (19.33 ± 1.75). Sequencing of typical bands revealed common fungal genera - Alternaria, Penicillium, Fusarium, Rhizopus, Isaria, and Metarhizium. Profiles of the S soil were enriched by bands of rumen-born anaerobic fungi (Neocallimastix, Cyllamyces) occurring mainly in profiles of excrements, where relatively low band richness (14.33 ± 1.15) was observed. The increasing level of cattle disturbance induced an increase in the biomass of complex fungal community over the three-year experimental period from 3.39 ± 2.11 (mean ± SD) nmol of fungal PLFA per gram of the C soil to 5.87 ± 3.16 in the M soil and 9.21 ± 4.69 in the S soil. Concentrations of soil N_tot and C_org were evaluated as the parameters significantly correlating with biomass as well as composition of the fungal community.     

Development of soil microbial communities during tallgrass prairie restoration

Soil microbial communities were examined in a chronosequence of four different land-use treatments at the Konza Prairie Biological Station, Kansas. The time series comprised a conventionally tilled cropland (CTC) developed on former prairie soils, two restored grasslands that were initiated on former agricultural soils in 1998 (RG₉₈) and 1978 (RG₇₈), and an annually burned native tallgrass prairie (BNP), all on similar soil types. In addition, an unburned native tallgrass prairie (UNP) and another grassland restored in 2000 (RG₀₀) on a different soil type were studied to examine the effect of long-term fire exclusion vs. annual burning in native prairie and the influence of soil type on soil microbial communities in restored grasslands. Both 16S rRNA gene clone libraries and phospholipid fatty acid analyses indicated that the structure and composition of bacterial communities in the CTC soil were significantly different from those in prairie soils. Within the time series, soil physicochemical characteristics changed monotonically. However, changes in the microbial communities were not monotonic, and a transitional bacterial community formed during restoration that differed from communities in either the highly disturbed cropland or the undisturbed original prairie. The microbial communities of RG₉₈ and RG₀₀ grasslands were also significantly different even though they were restored at approximately the same time and were managed similarly; a result attributable to the differences in soil type and associated soil chemistry such as pH and Ca. Burning and seasonal effects on soil microbial communities were small. Similarly, changing plot size from 300 m² to 150 m² in area caused small differences in the estimates of microbial community structure. In conclusion, microbial community structure and biochemical properties of soil from the tallgrass prairie were strongly impacted by cultivation, and the microbial community was not fully restored even after 30 years.     

喀斯特生态系统中乔木和灌木林根际土壤微生物生物量及其多样性的比较

我国喀斯特区域面积分布较广,而喀斯特生态系统的退化已成为当前西南地区面临的严重的生态问题。本研究选取贵州中部两种不同植被类型的生态系统—乔木林和灌木林,以乔木林中的白栎、园果化香和灌木林中的火棘、竹叶椒等主要优势树种为对象,研究不同的植物树种对根际土壤微生物生物量及其细菌群落结构的影响。结果显示:乔木林系统中根际土壤微生物生物量碳、氮显著性高于灌木林,植物的根际效应在乔木林中表现更为显著;同时乔木林中的优势树种通过根系分泌物的作用显著提高根际土壤细菌多样性指数,而灌木林中优势树种的根际土壤微生物量及多样性均未表现出明显的根际效应。因此,植被的演替通过改变土壤微生物的特性影响植物-微生物-土壤之间的物质和能量循环,进一步影响喀斯特生态系统的稳定和健康功能。     

健康与罹患青枯病的番茄土壤细菌群落特征比较

应用实时荧光定量PCR及MiSeq高通量测序技术,全面地研究了连作番茄田块中健康与感染青枯病植株周围土体及根际土壤细菌群落结构和组成.结果表明:健康番茄土体土壤的pH及全碳含量显著高于感病番茄土体土壤;土体及根际土壤的细菌群落结构和组成明显不同于感病番茄土体及根际土壤细菌群落.与感病番茄根际相比,健康番茄根际细菌的数量...     

林火预测预报研究综述

在危害森林的诸因子中,火灾是一种最具破坏性的灾害,各林业先进国家都非常重视林火研究工作。在世界范围内,林火预测预报的应用已成为越来越多的有林国家林火管理的有力工具。林火预测预报从二十世纪20年代迄今,已有80余年的历史,在世界各国发展很快。林火预测预报分为火险天气预报、林火发生预报和林火行为预报。林火预报研究中的关键问题是选择主要的林火因子和合适的林火预测预报方法。     

潮土中残留小麦和玉米秸秆养分含量差异及与微生物群落组成的关系

黄淮海平原典型潮土上小麦和玉米收获后的秸秆往往直接还田,但驱动它们在不同质地潮土(砂质、壤质、黏质)中分解的微生物是否与残留秸秆养分含量有关尚不清楚.本研究基于尼龙网袋法,通过10个月的田间培育试验,监测秸秆分解率、残留秸秆养分含量及微生物群落组成,评估各指标在秸秆类型和土壤质地之间的差异,探究残留秸秆养分与微生物群落...     

Bacterial communities in soybean rhizosphere in response to soil type, soybean genotype, and their growth stage

Three experiments were conducted in this study in order to investigate the impacts of soil type, soybean genotype, and the reproductive growth stage on bacterial communities in the soybean rhizosphere. Communities were evaluated by principal component analysis of denaturing gradient gel electrophoresis (DGGE) banding patterns and sequencing of partial 16S rDNA polymerase chain reaction (PCR) amplicons. A pot experiment analyzing three soybean genotypes grown in two different types of soil (Black soil and Dark Brown soil) indicated that soil type was the major factor in influencing the bacterial communities in the soybean rhizosphere, with a more significant effect observed in the Black soil samples than in the Dark Brown soil samples. A field experiment was conducted in Dark Brown soil using three soybean genotypes, and the results gleaned from both pot and field experiments indicated that bacterial communities in the soybean rhizosphere changed with growth stages, and higher number of DGGE bands observed in early reproductive growth stages, while surprisingly, a significant impact of genotype on the bacterial communities was not observed in these experiments. However, a plate culture experiment targeting the culturable bacterial communities detected a remarkable difference in the community structures of the rhizosphere between the two soybean genotypes, suggesting that a small portion of the total bacteria was influenced by genotype. Sequence analysis of DGGE bands indicated that bacterial phyla of Proteobacteria, Actinobacteria, Bacteroidetes, Nitrospirae, Firmicutes, Verrucomicrobia and Acidobacteria commonly inhabit the soybean rhizosphere.     

Frequent addition of wheat straw residues to soil enhances carbon mineralization rate

In many ecosystems, residues are added frequently to soil, in the form of root turnover and litter fall. However, in most studies on residue decomposition, residues are added once and there are few studies that have investigated the effect of frequent residue addition on C mineralization and N dynamics. To close this knowledge gap, we mixed mature wheat residue (C/N 122) into soil at a total rate of 2% w/w once at the start (R1×), every 16 days (R4×), every 8 days (R8×) or every 4 days (R16×). Un-amended soil served as control. All treatments were mixed every 4 days. Soil respiration was measured continuously over the 80-day incubation. Inorganic N, K₂SO₄-extractable C and N, chloroform-labile C and N (as an estimate of microbial biomass C and N), soil pH and microbial community composition were assessed every 16 days. Increasing frequency of residue addition increased C mineralization per g residue. Compared to R1×, cumulative respiration per g residue at the end of the incubation (day 80) was increased by 57, 82 and 92% in R4×, R8× and R16×, respectively. The largest differences in soil respiration per g residue occurred in the first 30 days. Despite large increases in cumulative respiration, frequent residue addition did not affect inorganic N or K₂SO₄-extractable N concentrations, chloroform-labile C and N or soil pH. Compared to the control, all residue treatments resulted in increases in chloroform-labile C and N and soil pH but decreased inorganic and K₂SO₄-extractable N. Microbial community composition was affected by residue addition, however there were no consistent differences among residue treatments. It is concluded that experiments with single residue additions may underestimate residue decomposition rates in the field. The increased C mineralization caused by frequent residue additions does not appear to be due to an increased microbial biomass or changes in microbial community composition, but rather to increased C mineralization per unit biomass.     

案头牡丹农艺指标与基质配方理化指标关系的研究

采用主成分分析的方法对牡丹农艺指标进行了简化,然后采用典型相关分析法对反映案头牡丹生长状况的一组指标与反映基质理化性质状况的另一组指标进行分析。结果表明案头牡丹的农艺指标与基质理化指标相关极显著,两组指标之间的相关主要是由株花蕾数与容重有显著相关引起的。统计分析提示案头牡丹与基质配方理化性质的差别主要是由于容重造成的。案头牡丹的开花朵数主要与电导率的高低有关。     

喀斯特小流域土壤饱和导水率垂直分布特征

土壤剖面饱和导水率(Saturated hydraulic conductivity,Ks)的垂直分布对土壤水文过程有极其重要的影响,但在地质背景特殊的喀斯特地区其研究还相对匮乏。通过测定典型喀斯特小流域内23个土壤剖面(0~10、10~20、20~30、30~50、50~70、70~100 cm)土壤Ks及土壤碎石含量(Rock fragment content,RC)、容重(Bulk density,BD)、毛管孔隙度(Capillary porosity,CP)、非毛管孔隙度(Non-capillary porosity,NCP)、土壤有机碳(Soil organic carbon,SOC)等土壤性质,并结合各样点的坡位(Slope position,SP)、坡度(Slope gradient,SG)、坡向(Slope aspect,SA)、裸岩率(Bare rock,BR)、土地利用类型(Land-use type,LU)等环境因素,应用偏相关分析和典范对应分析(CCA)的方法,研究了喀斯特小流域Ks的垂直分布特征及其主要影响因素。结果表明,Ks随土壤深度的增加而减小并可用对数函数模拟(R2=0.848)。20~100 cm各层Ks变幅较小且变异接近,因此在水文模型中可用20~30 cm土壤Ks代替深层。土壤性质中,RC与Ks的相关系数(0.484)最大。环境因素对Ks垂直分布的影响依次为SPSGSALUBR。由此可知,RC是影响Ks最重要的土壤性质,而SP则是影响Ks垂直分布最重要的环境因素。该结果有利于弄清喀斯特地区降雨入渗规律及其主要影响因素,为小流域植被恢复及水文模型的构建提供科学依据。