石油污染盐碱土壤棉花根际微生物与石油烃降解关系

【目的】探讨棉花(Gossypium spp.)生长对石油烃(TPH)污染盐碱土壤微生物群落结构的影响,揭示根际微生物与TPH降解的相关关系。【方法】利用磷脂脂肪酸(PLFA)方法解析根际土壤活性微生物群落随棉花生长的动态变化特征。【结果】根际土壤先后出现了21种PLFAs,包括：饱和脂肪酸(SAT),标识除放线菌之外的细菌;甲基支链末端型饱和脂肪酸(TBSAT),标识除放线菌之外的革兰氏阳性(G+)细菌;标识真菌的多不饱和脂肪酸(PUFA);标识放线菌的甲基支链中间型饱和脂肪酸(MBSAT);标识革兰氏阴性(G?)细菌的单不饱和脂肪酸(MONO)和环丙基脂肪酸(CYCLO)。棉花根际与未栽种棉花的对照(CK)相比,根际土壤微生物PLFAs种类在苗期、蕾期、吐絮期分别增加了100%、83.3%、20.0%,生物量分别增加了53.9%、6.60倍和60.7%;土壤TPH降解率分别提高13.0%、28.0%和30.6%。相关性分析表明：根际土壤TPH降解与根际土壤微生物总生物量具有低度正相关关系(|r|=0.5),但与a14:0、a16:0、i15:0标记的G+细菌生物量高度正相关(|r|≥0.8)。【结论】棉花生长对石油污染盐碱土壤活性微生物群落结构具有显著(p<0.05)的影响,且加速了土壤TPH的降解。该结果将为今后更好地开展石油污染盐碱土壤的生物修复技术研究提供理论依据。     

磷脂脂肪酸法在土壤微生物群落分析中的应用

土壤微生物群落的组成一直是土壤学、微生物学和生态学研究的热点问题。我国在这方面的研究处于国际前列,越来越多的研究成果在国际重要刊物上发表。而磷脂脂肪酸(PLFA)法在土壤微生物群落分析中占有举足轻重的地位,国内外学者都热衷于使用该方法。但是PLFA法的使用仍存在一些不足的地方,需要研究学者们慎重使用。本文综述了国际上相关研究,概述了PLFA方法使用的发展历史,应用及挑战。总结了使用和数据解读时需要注意的问题,整理了PLFA法相关的生物标记以及与新方法结合的设想,方便以后研究的开展。     

Enhancement of the microbial community biomass and diversity during air sparging bioremediation of a soil highly contaminated with kerosene and BTEX

In order to obtain insights in complexity shifts taking place in natural microbial communities under strong selective pressure, soils from a former air force base in the Czech Republic, highly contaminated with jet fuel and at different stages of a bioremediation air sparging treatment, were analyzed. By tracking phospholipid fatty acids and 16S rRNA genes, a detailed monitoring of the changes in quantities and composition of the microbial communities developed at different stages of the bioventing treatment progress was performed. Depending on the length of the air sparging treatment that led to a significant reduction in the contamination level, we observed a clear shift in the soil microbial community being dominated by Pseudomonads under the harsh conditions of high aromatic contamination to a status of low aromatic concentrations, increased biomass content, and a complex composition with diverse bacterial taxonomical branches. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. The online version of an erratum to this article can be found at http://dx.doi.org/. An erratum to this article can be found at     

Sheep-urine-induced changes in soil microbial community structure

Soil microbial communities play an important role in nutrient cycling and nutrient availability, especially in unimproved soils. In grazed pastures, sheep urine causes local changes in nutrient concentration which may be a source of heterogeneity in microbial community structure. In the present study, we investigated the effects of synthetic urine on soil microbial community structure, using physiological (community level physiological profiling, CLPP), biochemical (phospholipid fatty acid analysis, PLFA) and molecular (denaturing gradient gel electrophoresis, DGGE) fingerprinting methods. PLFA data suggested that synthetic urine treatment had no significant effect on total microbial (total PLFA), total bacterial or fungal biomass; however, significant changes in microbial community structure were observed with both PLFA and DGGE data. PLFA data suggested that synthetic urine induced a shift towards communities with higher concentrations of branched fatty acids. DGGE banding patterns derived from control and treated soils differed, due to a higher proportion of DNA sequences migrating only to the upper regions of the gel in synthetic urine-treated samples. The shifts in community structure measured by PLFA and DGGE were significantly correlated with one another, suggesting that both datasets reflected the same changes in microbial communities. Synthetic urine treatment preferentially stimulated the use of rhizosphere-C in sole-carbon-source utilisation profiles. The changes caused by synthetic urine addition accounted for only 10-15% of the total variability in community structure, suggesting that overall microbial community structure was reasonably stable and that changes were confined to a small proportion of the communities.     

木焦油污染土壤中微生物特性的空间变异性研究

运用地统计学方法研究了木焦油污染土壤中微生物量、微生物群落结构、微生物活性等的空间变异特征.分别采用26种主要磷脂类脂肪酸(PLFA)的总含量(totPLFA)、PLFA的第一主成分和第二主成分(PLFA PC1和PC2)以及土壤培养过程中CO₂-C的累积释放量(Cre)来表征土壤中的微生物量、微生物群落结构以及微生物活性.结果表明,多数微生物特性指标均存在不同程度的空间自相关性,其半变异函数曲线可用带块金效应的球状模型进行拟合.变量的空间相关距离在2.50～16.60 m之间.PLFA PC1、totPLFA和Cre均具有较强的空间依赖性,其相对结构变差（由结构性因素引起的空间变异）分别为82.3%、79.6%和64.7%,而PLFA PC2 不存在明显的空间依赖性.克立格空间插值图表明,样地中存在几处微生物相对密集分布且代谢活性较高的区域,其中优势微生物菌群是由PLFAs 16:1ω7t,cy17:0,18:1ω7 和cy19:0所表征的革兰氏阴性细菌.土壤中主要污染物多环芳烃含量和空间分布是影响微生物特性空间分布格局的重要因素之一.     

杉木人工林土壤微生物群落结构特征

采用氯仿熏蒸法、稀释平板法和磷脂脂肪酸(phospholipid fatty acid,PLFA)方法,分析了常绿阔叶林转变成杉木人工林后土壤微生物种群数量和群落结构的变化特征.结果表明:常绿阔叶林转变为杉木人工林后,林地土壤的微生物生物量碳、可培养细菌和放线菌数降低.杉木人工林地总PLFAs、细菌PLFAs、真菌PLFAs比常绿阔叶林分别降低了49.4％、52.4％和46.6％,革兰氏阳性和阴性细菌PLFAs远低于常绿阔叶林.杉木人工林根际土壤微生物生物量碳、可培养细菌和放线菌数显著高于杉木人工林林地土壤,根际土壤中总PLFAs、细菌PLFAs、革兰氏阳性和阴性细菌PLFAs的含量也高于林地土壤,但真菌PLFAs和细菌PLFAs之比却低于林地土壤.对土壤微生物群落结构进行主成分分析发现,第1主成分和第2主成分共解释了土壤微生物群落结构变异的78.2％.表明常绿阔叶林与杉木人工林土壤的微生物群落结构间存在差异.     

Effect of stand age on soil microbial community structure in wolfberry (Lycium barbarum L.) fields

Soil physicochemical properties and microbes are essential in terrestrial ecosystems through their role in cycling mineral compounds and decomposing organic matter. This study examined the effect of stand age on soil physicochemical properties and microbial community structure in wolfberry (Lycium barbarum L.) fields, in order to reveal the mechanism of soil degradation due to long-term stand of L. barbarum. The objective of the study was achieved by phospholipid fatty acid (PLFA) biomarker analysis of soil samples from L. barbarum fields in Zhongning County, Ningxia Province—the origin of L. barbarum. Five stand ages of L. barbarum were selected, < 1, 3, 6, 9, and 12 years (three plots each). The results showed that soil bulk density increased slightly with increasing stand age, while no clear trend was observed in soil pH or total salinity. As the stand age increased, soil organic matter and nutrients first increased before decreasing, with the highest levels being found in year 9. There was an amazing variety of PLFA biomarkers in soil samples at different stand ages. The average concentrations of total, bacterial, fungal, and actinomycete PLFAs in the surface soil initially decreased and then increased, before decreasing with the stand age in summer. The PLFA concentrations of major microbial groups were highest in year 9, with the total PLFA concentrations being 32.97% and 10.67% higher than those in years < 1 and 12, respectively. Higher microbial PLFA concentrations were detected in summer relative to autumn and in the surface relative to the subsurface soil. The highest ratios of Gram-positive to Gram-negative bacterial (G^?/G⁺) and fungal to bacterial (F/B) PLFAs were obtained in year 6, on average 76.09% higher than those at the other four stand ages. The soil environment was most stable in year 6, with no differences between other stand ages. Therefore, soil microbial community structure was strongly influenced by the stand age in year 6 only. The effect of stand age on soil G^?/G⁺ and microbial community structure varied with season and depth; there was little effect for F/B in the 20–40 cm soil layer. Principal component analysis revealed no correlations between microbial PLFA concentrations and total salinity in the soil; negative correlations were noted between soil pH and F/B in summer (P < 0.01), as well as between soil pH and fungal PLFA in autumn (P < 0.05). Moreover, microbial PLFA concentrations were correlated with soil organic matter (mean R = 0.7725), total nitrogen (mean R = 0.8296), total phosphorus (mean R = 0.8175), available nitrogen (mean R = 0.7458), and available phosphorus (mean R = 0.7795) (P < 0.01). On the whole, the soil ecosystem was most stable in year 6, while soil organic matter, nutrients, and microbial PLFA concentrations were maximal in year 9; thereafter, soil fertility indices and microbial concentrations decreased and soil quality declined gradually as the stand age increased. Therefore, farmers should reduce the application rate of fertilizers, especially compound or mixed fertilizers, in L. barbarum fields; organic or bacterial manure can be applied increasingly to improve the soil environment and prolong the economic life of L. barbarum.     

有机无机缓释复合肥对土壤微生物量碳、氮和群落结构的影响

研究缓释复合肥不同用量对土壤微生物量碳、氮和群落结构多样性的影响,在农业生产上广泛应用缓释复合肥有着重要意义。试验采用室内长期恒温培养和磷脂脂肪酸法,以化肥和普通复合肥适量施用养分量为对比,研究缓释复合肥适量、高量和超高量施肥水平对土壤微生物PLFA含量的影响规律。结果表明,SRF1、SRF2、SRF3(缓释复合肥适量、高量和超高量)较CK(不施肥)和CF1(化肥适量)显著增加土壤微生物量碳,且较CK、CF1和CCF1(普通复合肥适量)显著增加土壤微生物量氮。土壤微生物量碳、氮随着缓释复合肥施肥水平的增加而增加,但没有随着施肥水平的倍量增加而倍量增加,且SRF2和SRF3无显著差异。缓释复合肥(SRF1、SRF2和SRF3)较CK、CF1和CCF1增加土壤PLFA的种类和含量,且总PLFA含量增加7.4%—26.7%、17.6%—38.7%和12.8%—33.0%,3个施肥水平以SRF2作用效果最好,总PLFA含量最高,分别较SRF1和较SRF3高16.4%和17.9%。土壤细菌、放线菌、革兰氏阳性菌和革兰氏阴性菌PLFA含量以SRF1和SRF2显著高于CF1和CCF1。主成分分析和聚类分析显示施肥处理分布较多PLFA的优势种群,SRF3与SRF1和SRF2的PLFA结构差别较大。综上认为,适量施肥水平以缓释复合肥较化肥和普通复合肥对土壤微生物的作用显著,其中缓释复合肥3个施肥水平以高量施肥水平作用最好。     

鄱阳湖湿地土壤微生物群落结构沿地下水位梯度分异特征

为了揭示地下水位梯度对湿地土壤微生物群落的影响,在鄱阳湖典型碟形湖泊白沙湖洲滩湿地设置了200m×300m大样地,沿地下水位梯度划分4个样带(从湖岸到湖心依次为GT-A,GT-B,GT-C,GT-D),采集了不同梯度带的土壤样品,利用磷脂脂肪酸法分析其土壤微生物群落结构分异特征。结果表明,随着地下水位抬升,土壤pH和沙粒含量升高,而有机碳、容重、粘粒和粉粒含量降低。与地下水位最低的梯度(GT-A)相比,地下水位在地表上下波动(GT-D)时,土壤微生物量碳氮及其分配比例分别增加了2.82、4.30、5.77和7.15倍;土壤微生物总量、细菌生物量、放线菌生物量、革兰氏阳性细菌及革兰氏阴性细菌生物量分别增长了106.8%、117.2%、74.9%,107.9%和207.2%。洲滩地下水位梯度的升高增加了土壤微生物群落的环境压力,进而降低了其群落结构的多样性。土壤微生物群落结构组成与土壤pH、含水量、沙粒含量以及碳氮比呈显著相关关系,而土壤微生物商则主要受pH和土壤质地的影响。以上结果表明地下水位梯度所引起的土壤微环境变化对微生物量、土壤有机碳周转和群落结构均产生了深刻影响。     

Seasonal changes in the microbial community of a salt marsh, measured by phospholipid fatty acid analysis

Microbial activity within the environment can have distinct geochemicaleffects, and so changes in a microbial community structure can result ingeochemical change. We examined seasonal changes in both the microbialcommunityand the geochemistry of an inter-tidal salt marsh in north-west England tocharacterise biogeochemical processes occurring at this site.Phospholipid fatty acid (PLFA) analysis of sediment samples collected atmonthly intervals was used to measure seasonal changes in microbial biomass andcommunity structure. The PLFA data were analysed using multivariate techniques(Ward's method and the Mahalanobis distance metric), and we show that the useofthe Mahalanobis distance metric improves the statistical analysis by providingdetailed information on the reasons samples cluster together and identifyingthedistinguishing features between the separate clusters. Five clusters of likesamples were defined, showing differences in the community structure over thecourse of a year.At all times, the microbial community was dominated by PLFA associated withaerobic bacteria, but this was most pronounced in summer (August). Theabundanceof branched fatty acids, a measure of the biomass of anaerobes, started toincrease later in the year than did those associated with aerobes and thefungalbiomarker 18:26 showed a brief late-summer peak.The salt marsh remained mildly oxic throughout the year despite the increase inmicrobial respiration, suggested by the large increases in the abundance ofPLFA, in the warmer months. The conditions therefore remained most favourablefor aerobic species throughout the year, explaining their continual dominanceatthis site. However, as the abundance of PLFA synthesised by anaerobesincreased,increases in dissolved Mn concentrations were observed, which we suggest weredue to anaerobic respiration of Mn(IV) to Mn(II). Overall, the geochemicalconditions were consistent with the microbial community structure and changeswithin it.     

培养条件下二氯喹啉酸对土壤微生物群落结构的影响

为探讨除草剂二氯喹啉酸对土壤微生物群落结构的影响,在25℃黑暗培养条件下,采用磷脂脂肪酸法(Phospholipid Fatty Acid,PLFA)分析了二氯喹啉酸(0对照、83.3和166.6μg/kg干土)处理对淹水和不淹水水田土壤微生物群落结构的影响。结果表明,在实验处理的60 d内,不淹水水田土壤在83.3μg/kg二氯喹啉酸处理后对土壤微生物生物总量、细菌生物量及真菌生物量有抑制作用,而166.6μg/kg二氯喹啉酸处理后各生物量指标均显著增加,但是两种浓度处理均使土壤真菌/细菌比值下降,说明二氯喹啉酸处理会使不淹水水田土壤微生物稳定性下降;淹水水田土壤在83.3μg/kg二氯喹啉酸处理下,对土壤微生物生物总量和真菌生物量有促进作用,对细菌生物量有一定的抑制作用,166.6μg/kg二氯喹啉酸处理对土壤微生物生物总量、细菌及真菌生物量均有促进作用,从真菌/细菌比值上看,不同浓度处理与对照无显著差异,说明二氯喹啉酸对淹水生物稳定性无明显干扰。PLFA主成分分析表明,不同浓度处理的两种土壤微生物群落均以含14∶0、15∶0、16∶0和18∶2n6c的微生物为优势种群。     

红壤侵蚀区芒萁对土壤微生物群落结构的影响

土壤微生物是反映土壤质量状况的重要指标,研究侵蚀地植被恢复后土壤微生物群落结构的变化对深入认识土壤质量的演变具有重要意义。对比分析了未治理地(Y0)、治理13年(Y13)和31年(Y31)的马尾松林(Pinus massoniana)林下芒萁(Dicranopteris dichotoma)覆盖地(NRd)、去除芒萁覆盖地(Rd)与林下裸地(CK)土壤微生物生物量和群落结构差异,结果表明:林下裸地土壤微生物生物量碳(MBC)、微生物生物量氮(MBN)和总微生物磷脂脂肪酸量(总PLFAs)的含量均显著低于芒萁覆盖地,且去除芒萁4个月后,MBC和总PLFAs均有降低趋势,表明芒萁覆盖对土壤微生物生物量具有重要影响;林下芒萁覆盖地土壤革兰氏阳性菌(GP)、革兰氏阴性菌(GN)、丛植菌根真菌(VAM)、真菌(Fungi)、放线菌(ACT)的PLFAs含量显著高于林下裸地(Y13例外),去除芒萁4个月后,各值均有有接近林下裸地的趋势;芒萁覆盖地真菌/细菌的比值(F/B)均显著高于林下裸地(P0.05),芒萁覆盖地革兰氏阳性菌/革兰氏阴性菌的比值(GP/GN)、饱和直链脂肪酸/单不饱和脂肪酸的比值(sat/mono)和(cy17:0+cy19:0ω8c)/(16:1ω7c+18:1ω7c)(cy/pre)显著小于林下裸地(P0.05),去除芒萁4个月后,芒萁覆盖地土壤cy/pre显著升高(P0.05)(Y13例外),意味着芒萁覆盖地土壤生态系统更稳定,土壤的养分可利用性更高,微生物生物量和群落结构更丰富,活性更强;皮尔逊相关分析和冗余分析发现,土壤理化性质与土壤微生物生物量和群落结构关系密切,土壤C/N、p H和氮素水平是调控芒萁覆盖下土壤微生物生物量和群落结构的主要生态因子。     

增温对长白山苔原土壤微生物群落结构的影响

研究土壤微生物群落结构对温度升高的响应,对预测气候变化条件下土壤微生物以及土壤养分循环具有重要意义。采用开顶箱(OTC,Open-top chamber)增温方法对长白山苔原土壤进行连续两个生长季(6—9月)增温处理,结果表明:增温使土壤磷脂脂肪酸(PLFA,Phospholipid fatty acid)总量降低了16.1%,革兰氏阳性菌/革兰氏阴性菌比值(G+/G-)升高21.2%。增温与对照条件下的G+、G-、细菌、真菌的PLFAs相对含量和真菌/细菌比值在统计上无显著差异,除真菌与G-外,其它指标均存在明显的季节波动。增温与对照条件下,细菌、G+、G+/G-和PLFA总量在土壤温度较高的7、8月份较温度较低的9月份高,真菌/细菌比值则在9月份温度较低时达到最大值。主成分分析表明,整个生长季代表真菌和G-的脂肪酸相对变化较明显。冗余分析(RDA,Redundancy analysis)表明,G+/G-比值与土壤温度呈正相关关系,土壤含水量与PLFA总量呈负相关关系,表明增温直接或间接导致G+/G-比值和PLFA总量变化,改变了土壤微生物的群落结构。     

Plant community and soil conditions individually affect soil microbial community assembly in experimental mesocosms

Soils harbor large, diverse microbial communities critical for local and global ecosystem functioning that are controlled by multiple and poorly understood processes. In particular, while there is observational evidence of relationships between both biotic and abiotic conditions and microbial composition and diversity, there have been few experimental tests to determine the relative importance of these two sets of factors at local scales. Here, we report the results of a fully factorial experiment manipulating soil conditions and plant cover on old‐field mesocosms across a latitudinal gradient. The largest contributor to beta diversity was site‐to‐site variation, but, having corrected for that, we observed significant effects of both plant and soil treatments on microbial composition. Separate phyla were associated with each treatment type, and no interactions between soil and plant treatment were observed. Individual soil characteristics and biotic parameters were also associated with overall beta‐diversity patterns and phyla abundance. In contrast, soil microbial diversity was only associated with site and not experimental treatment. Overall, plant community treatment explained more variation than soil treatment, a result not previously appreciated because it is difficult to dissociate plant community composition and soil conditions in observational studies across gradients. This work highlights the need for more nuanced, multifactorial experiments in microbial ecology and in particular indicates a greater focus on relationships between plant composition and microbial composition during community assembly.     

Root controls on soil microbial community structure in forest soils

We assessed microbial community composition as a function of altered above- and belowground inputs to soil in forest ecosystems of Oregon, Pennsylvania, and Hungary as part of a larger Detritus Input and Removal Treatment (DIRT) experiment. DIRT plots, which include root trenching, aboveground litter exclusion, and doubling of litter inputs, have been established in forested ecosystems in the US and Europe that vary with respect to dominant tree species, soil C content, N deposition rate, and soil type. This study used phospholipid fatty-acid (PLFA) analysis to examine changes in the soil microbial community size and composition in the mineral soil (0–10 cm) as a result of the DIRT treatments. At all sites, the PLFA profiles from the plots without roots were significantly different from all other treatments. PLFA analysis showed that the rootless plots generally contained larger quantities of actinomycete biomarkers and lower amounts of fungal biomarkers. At one of the sites in an old-growth coniferous forest, seasonal changes in PLFA profiles were also examined. Seasonal differences in soil microbial community composition were greater than treatment differences. Throughout the year, treatments without roots continued to have a different microbial community composition than the treatments with roots, although the specific PLFA biomarkers responsible for these differences varied by season. These data provide direct evidence that root C inputs exert a large control on microbial community composition in the three forested ecosystems studied.     

Fatty acid methyl ester (FAME) profiles as measures of soil microbial community structure

Analysis of fatty acid methyl ester (FAME) profiles extracted from soils is a rapid and inexpensive procedure that holds great promise in describing soil microbial community structure without traditional reliance on selective culturing, which seems to severely underestimate community diversity. Interpretation of FAME profiles from environmental samples can be difficult because many fatty acids are common to different microorganisms and many fatty acids are extracted from each soil sample. We used principal components (PCA) and cluster analyses to identify similarities and differences among soil microbial communities described using FAME profiles. We also used PCA to identify particular FAMEs that characterized soil sample clusters. Fatty acids that are found only or primarily in particular microbial taxa-marker fatty acids-were used in conjunction with these analyses. We found that the majority of 162 soil samples taken from a conventionally-tilled corn field had similar FAME profiles but that about 20% of samples seemed to have relatively low, and that about 10% had relatively high, bacterial:fungal ratios. Using semivariance analysis we identified 21:0 iso as a new marker fatty acid. Concurrent use of geostatistical and FAME analyses may be a powerful means of revealing other potential marker FAMEs. When microbial communities from the same samples were cultured on R2A agar and their FAME profiles analyzed, there were many differences between FAME profiles of soil and plated communities, indicating that profiles of FAMEs extracted from soil reveal portions of the microbial community not culturable on R2A. When subjected to PCA, however, a small number of plated communities were found to be distinct due to some of the same profile characteristics (high in 12:0 iso, 15:0 and 17:1 ante A) that identified soil community FAME profiles as distinct. Semivariance analysis indicated that spatial distributions of soil microbial populations are maintained in a portion of the microbial community that is selected on laboratory media. These similarities between whole soil and plated community FAME profiles suggest that plated communities are not solely the result of selection by the growth medium, but reflect the distribution, in situ, of the dominant, culturable soil microbial populations.