Diversity, composition, and geographical distribution of microbial communities in California salt marsh sediments

The Pacific Estuarine Ecosystem Indicators Research Consortium seeks to develop bioindicators of toxicant-induced stress and bioavailability for wetland biota. Within this framework, the effects of environmental and pollutant variables on microbial communities were studied at different spatial scales over a 2-year period. Six salt marshes along the California coastline were characterized using phospholipid fatty acid (PLFA) analysis and terminal restriction fragment length polymorphism (TRFLP) analysis. Additionally, 27 metals, six currently used pesticides, total polychlorinated biphenyls and polycyclic aromatic hydrocarbons, chlordanes, nonachlors, dichlorodiphenyldichloroethane, and dichlorodiphenyldichloroethylene were analyzed. Sampling was performed over large (between salt marshes), medium (stations within a marsh), and small (different channel depths) spatial scales. Regression and ordination analysis suggested that the spatial variation in microbial communities exceeded the variation attributable to pollutants. PLFA analysis and TRFLP canonical correspondence analysis (CCA) explained 74 and 43% of the variation, respectively, and both methods attributed 34% of the variation to tidal cycles, marsh, year, and latitude. After accounting for spatial variation using partial CCA, we found that metals had a greater effect on microbial community composition than organic pollutants had. Organic carbon and nitrogen contents were positively correlated with PLFA biomass, whereas total metal concentrations were positively correlated with biomass and diversity. Higher concentrations of heavy metals were negatively correlated with branched PLFAs and positively correlated with methyl- and cyclo-substituted PLFAs. The strong relationships observed between pollutant concentrations and some of the microbial indicators indicated the potential for using microbial community analyses in assessments of the ecosystem health of salt marshes.     

Spatial and temporal variations in salt marsh microorganisms of the Wadden Sea

Salt marshes exist at the interface of the marine and the terrestrial system. Shore height differences and associated variations in inundation frequency result in altered abiotic conditions, plant communities, and resource input into the belowground system. These factors result in three unique zones, the upper salt marsh (USM), the lower salt marsh (LSM), and the pioneer zone (PZ). Marine detritus, such as micro‐ and macroalgae, is typically flushed into the PZ daily, with storm surges moving both salt marsh detritus and marine detritus into higher salt marsh zones. Microbial assemblages are essential for the decomposition of organic matter and have been shown to sensitively respond to changes in abiotic conditions such as oxygen supply and salinity. However, temporal and spatial dynamics of microbial communities of Wadden Sea salt marshes received little attention. We investigated the dynamics of soil microbial communities across horizontal (USM, LSM, and PZ), vertical (0–5 and 5–10‐cm sediment depth), and temporal (spring, summer, and autumn) scales in the Wadden Sea salt marsh of the European North Atlantic coast using phospholipid fatty acid (PLFA) analysis. Our results show strong spatial dynamics both among salt marsh zones and between sediment depths, but temporal dynamics to be only minor. Despite varying in space and time, PLFA markers indicated that bacteria generally were the dominant microbial group across salt marsh zones and seasons, however, their dominance was most pronounced in the USM, whereas fungal biomass peaked in the LSM and algal biomass in the PZ. Only algal markers and the stress marker monounsaturated to saturated fatty acid ratio responded to seasonality. Overall, therefore, the results indicate remarkable temporal stability of salt marsh microbial communities despite strong variability in abiotic factors.     

Relationships between Sediment Microbial Communities and Pollutants in Two California Salt Marshes

Salt marshes are important ecosystems whose plant and microbial communities can alter terrestrially derived pollutants prior to coastal water discharge. However, knowledge regarding relationships between anthropogenic pollutant levels and salt marsh microbial communities is limited, and salt marshes on the West Coast of the United States are rarely examined. In this study, we investigated the relationships between microbial community composition and 24 pollutants (20 metals and 4 organics) in two California salt marshes. Multivariate ordination techniques were used to assess how bacterial community composition, as determined by terminal restriction fragment length polymorphism and phospholipid fatty acid analyses, was related to pollution. Sea urchin embryo toxicity measurements and plant tissue metabolite profiles were considered two other biometrics of pollution. Spatial effects were strongly manifested across marshes and across channel elevations within marshes. Utilizing partial canonical correspondence analysis, an ordination technique new to microbial ecology, we found that several metals were strongly associated with microbial community composition after accounting for spatial effects. The major patterns in plant metabolite profiles were consistent with patterns across microbial community profiles, but sea urchin embryo assays, which are commonly used to evaluate ecological toxicity, had no identifiable relationships with pollution. Whereas salt marshes are generally dynamic and complex habitats, microbial communities in these marshes appear to be relatively sensitive indicators of toxic pollutants.     

南亚热带3种人工林土壤微生物生物量和微生物群落结构特征

以我国南亚热带格木、红椎和马尾松人工林为对象,采用氯仿熏蒸浸提法和磷脂脂肪酸法(PLFA)分析了林地土壤微生物生物量和微生物群落结构组成.结果表明: 林分和季节因素均显著影响土壤微生物生物量、总PLFAs量、细菌PLFAs量和真菌PLFAs量,且干季林分下的土壤微生物生物量、总PLFAs量、单个PLFA量均大于雨季.红椎人工林土壤微生物生物量碳(MBC)和总PLFAs量最高,而格木人工林土壤微生物生物量氮(MBN)最高.土壤pH值对土壤丛枝菌根真菌(16:1ω5c)的影响达到极显著正相关水平.土壤总PLFAs量、革兰氏阳性菌(G⁺)以及腐生真菌(18:2ω6,9c)、革兰氏阳性菌/革兰氏阴性菌(G⁺/G^-)与土壤有机碳、全氮和全磷显著相关,表明土壤有机碳、全氮、全磷含量是影响该地区土壤微生物数量和种类的重要因素.外生菌根真菌(18:1ω9c)和丛枝菌根真菌与土壤碳氮比值呈极显著相关.     

Phospholipid Fatty Acid Composition and Heavy Metal Tolerance of Soil Microbial Communities along Two Heavy Metal-Polluted Gradients in Coniferous Forests

The effects of long-term heavy metal deposition on microbial community structure and the level of bacterial community tolerance were studied along two different gradients in Scandinavian coniferous forest soils. One was near the Harjavalta smelter in Finland, and one was at Ronnskar in Sweden. Phospholipid fatty acid (PLFA) analysis revealed a gradual change in soil microbial communities along both pollution gradients, and most of the individual PLFAs changed similarly to metal pollution at both sites. The relative quantities of the PLFAs br18:0, br17:0, i16:0, and i16:1 increased with increasing heavy metal concentration, while those of 20:4 and 18:2(omega)6, which is a predominant PLFA in many fungi, decreased. The fungal part of the microbial biomass was found to be more sensitive to heavy metals. This resulted in a decreased fungal/bacterial biomass ratio along the pollution gradient towards the smelters. The thymidine incorporation technique was used to study the heavy metal tolerance of the bacteria. The bacterial community at the Harjavalta smelter, exposed mainly to Cu deposition, exhibited an increased tolerance to Cu but not to Cd, Ni, and Zn. At the Ronnskar smelter the deposition consisting of a mixture of metals increased the bacterial community tolerance to all tested metals. Both the PLFA pattern and the bacterial community tolerance were affected at lower soil metal concentrations than were bacterial counts and bacterial activities. At Harjavalta the increased Cu tolerance of the bacteria and the change in the PLFA pattern of the microbial community were found at the same soil Cu concentrations. This indicated that the altered PLFA pattern was at least partly due to an altered, more metal-tolerant bacterial community. At Ronnskar, where the PLFA data varied more, a correlation between bacterial community tolerance and an altered PLFA pattern was found up to 10 to 15 km from the smelter. Farther away changes in the PLFA pattern could not be explained by an increased community tolerance to metals.     

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.     

磷脂脂肪酸方法在土壤微生物分析中的应用*

磷脂脂肪酸(PLYA)是活体微生物细胞膜的重要组分,不同类群的微生物能通过不同的生化途径合成不同的PLFA,部分PLFA可以作为分析微生物量和微生物群落结构等变化的生物标记。在土壤微生物分析中,越来越多地采用了PLFA方法。主要对PLFA方法在土壤微生物分析中的应用做一综述。     

Effect of salinity-altering pulsing events on soil organic carbon loss along an intertidal wetland gradient: a laboratory experiment

Salinity changes resulting from storm surge, tides, precipitation, and stormwater run-off are common in coastal wetlands. Soil microbial communities respond quickly to salinity changes, altering the rate of soil organic carbon (SOC) loss and associated biogeochemical processes. This study quantified the impact of salinity-altering pulses on SOC loss, defined as microbial respiration (CO₂ flux) at high and low tide, CH₄ flux, and dissolved OC (DOC) release, in 3 intertidal wetlands (Jacksonville, FL, USA). Intact soil cores from a freshwater tidal, brackish, and salt marsh were exposed to simulated tides and 3 salinity pulsing events during a 53-day laboratory experiment. Soil and water physio-chemical properties, nutrient release, and microbial indicators were measured. Microbial respiration was the dominate pathway of SOC loss (>97 %). Soil hydraulic conductivity was greater in brackish and salt marshes and was critical to overall soil respiration. High tide CO₂ flux was greatest in the freshwater marsh (58 % of SOC loss) and positively correlated with DOC concentration; low tide CO₂ flux was greatest in brackish and salt marshes (62 and 70 % of SOC loss, respectively) and correlated with NH₄ ⁺ and microbial biomass. The freshwater marsh was sensitive to brackish pulses, causing a 112 % increase in respiration, presumably from accelerated sulfate reduction and N-cycling. SOC loss increased in the salt marsh pulsed with freshwater, suggesting freshwater run-off may reduce a salt marsh’s ability to keep-pace with sea level rise. Increased inundation from storm surges could accelerate SOC loss in freshwater marshes, while decreasing SOC loss in brackish and salt marshes.     

Variation in Microbial Community Structure in Two Boreal Peatlands as Determined by Analysis of Phospholipid Fatty Acid Profiles

Analyses of phospholipid fatty acids (PLFAs) were used to assess variation in community structure and total microbial biomass in two boreal peatlands in Sweden. The total PLFA concentration in peat ranged from 0.16 to 7.0 nmol g of wet peat(sup-1) (median, 0.70 nmol g of wet peat(sup-1)). Principal-component analysis of PLFA data revealed that the degree of depth-related variation in PLFA composition was high among peatland habitats, with general differences between wet sites, with water tables within a few centimeters of the moss surface, and dry sites, with water tables >10 cm below the moss surface. However, variation in PLFA composition over the growing season was negligible. In the principal-component analyses, most PLFAs were determined to be parts of clusters of covarying fatty acids, suggesting that they originated in the same functional groups of microorganisms. Major clusters were formed by monounsaturated (typical of gram-negative eubacteria), terminally branched (gram-positive or anaerobic gram-negative eubacteria), methyl-branched and branched unsaturated (sulfate-reducing bacteria and/or actinomycetes), (omega)8 monounsaturated (methane-oxidizing bacteria), and polyunsaturated (eucaryotes) PLFAs. Within the clusters, PLFAs had rather distinct concentration-depth distributions. For example, PLFAs from sulfate-reducing bacteria and/or actinomycetes and those from methane-oxidizing bacteria had maximum concentrations slightly below and at the average water table depth, respectively.     

Cross-Site Soil Microbial Communities under Tillage Regimes: Fungistasis and Microbial Biomarkers

The exploitation of soil ecosystem services by agricultural management strategies requires knowledge of microbial communities in different management regimes. Crop cover by no-till management protects the soil surface, reducing the risk of erosion and nutrient leaching, but might increase straw residue-borne and soilborne plant-pathogenic fungi. A cross-site study of soil microbial communities and Fusarium fungistasis was conducted on six long-term agricultural fields with no-till and moldboard-plowed treatments. Microbial communities were studied at the topsoil surface (0 to 5 cm) and bottom (10 to 20 cm) by general bacterial and actinobacterial terminal restriction fragment length polymorphism (T-RFLP) and phospholipid fatty acid (PLFA) analyses. Fusarium culmorum soil fungistasis describing soil receptivity to plant-pathogenic fungi was explored by using the surface layer method. Soil depth had a significant impact on general bacterial as well as actinobacterial communities and PLFA profiles in no-till treatment, with a clear spatial distinction of communities (P < 0.05), whereas the depth-related separation of microbial communities was not observed in plowed fields. The fungal biomass was higher in no-till surface soil than in plowed soil (P < 0.07). Soil total microbial biomass and fungal biomass correlated with fungistasis (P < 0.02 for the sum of PLFAs; P < 0.001 for PLFA 18:2ω6). Our cross-site study demonstrated that agricultural management strategies can have a major impact on soil microbial community structures, indicating that it is possible to influence the soil processes with management decisions. The interactions between plant-pathogenic fungi and soil microbial communities are multifaceted, and a high level of fungistasis could be linked to the high microbial biomass in soil but not to the specific management strategy.     

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.     

氮肥添加对高寒藏嵩草(Kobresia tibetica)沼泽化草甸和土壤微生物群落的影响

以藏嵩草沼泽化草甸为研究对象,利用磷脂脂肪酸(PLFA)技术,研究连续6年N素添加对地上植被群落数量特征、土壤微生物群落结构的影响。结果表明:①藏嵩草沼泽化草甸群落生物量、枯枝落叶对施肥处理无明显响应,且莎草科植物对土壤氮素的吸收和利用率较低。②施肥增加了0-10 cm土壤微生物类群PLFAs丰富度尤其细菌和革兰氏阳性菌PLFAs,降低了10-20 cm PLFAs丰富度;③磷脂脂肪酸饱和脂肪酸/单烯不饱和脂肪酸、细菌PLFAs/真菌PLFAs的比值随土壤层次增加而增加;④0-10 cm土层革兰氏阳性菌、真菌PLFAs含量与pH、土壤速效磷、速效氮、土壤有机质显著正相关(P0.05或P0.01);10-20 cm土层,细菌、革兰氏阳性菌、真菌和总PLFAs含量与土壤有机质含量显著正相关(P0.05或P0.01)。表明藏嵩草沼泽化草甸微生物PLFAs含量和丰富度对施肥的响应存在明显的土层梯度效应,土壤微生物PLFAs含量和丰富度主要受表层土壤初始养分含量的影响。     

Phospholipid-Derived Fatty Acids and Quinones as Markers for Bacterial Biomass and Community Structure in Marine Sediments

Phospholipid-derived fatty acids (PLFA) and respiratory quinones (RQ) are microbial compounds that have been utilized as biomarkers to quantify bacterial biomass and to characterize microbial community structure in sediments, waters, and soils. While PLFAs have been widely used as quantitative bacterial biomarkers in marine sediments, applications of quinone analysis in marine sediments are very limited. In this study, we investigated the relation between both groups of bacterial biomarkers in a broad range of marine sediments from the intertidal zone to the deep sea. We found a good log-log correlation between concentrations of bacterial PLFA and RQ over several orders of magnitude. This relationship is probably due to metabolic variation in quinone concentrations in bacterial cells in different environments, whereas PLFA concentrations are relatively stable under different conditions. We also found a good agreement in the community structure classifications based on the bacterial PLFAs and RQs. These results strengthen the application of both compounds as quantitative bacterial biomarkers. Moreover, the bacterial PLFA- and RQ profiles revealed a comparable dissimilarity pattern of the sampled sediments, but with a higher level of dissimilarity for the RQs. This means that the quinone method has a higher resolution for resolving differences in bacterial community composition. Combining PLFA and quinone analysis as a complementary method is a good strategy to yield higher resolving power in bacterial community structure.     

Association of microbial community composition and activity with lead,chromium, and hydrocarbon contamination

Microbial community composition and activity were characterized in soil contaminated with lead (Pb), chromium (Cr), and hydrocarbons. Contaminant levels were very heterogeneous and ranged from 50 to 16,700 mg of total petroleum hydrocarbons (TPH) kg of soil(-1), 3 to 3,300 mg of total Cr kg of soil(-1), and 1 to 17,100 mg of Pb kg of soil(-1). Microbial community compositions were estimated from the patterns of phospholipid fatty acids (PLFA); these were considerably different among the 14 soil samples. Statistical analyses suggested that the variation in PLFA was more correlated with soil hydrocarbons than with the levels of Cr and Pb. The metal sensitivity of the microbial community was determined by extracting bacteria from soil and measuring [(3)H]leucine incorporation as a function of metal concentration. Six soil samples collected in the spring of 1999 had IC(50) values (the heavy metal concentrations giving 50% reduction of microbial activity) of approximately 2.5 mM for CrO(4)2- and 0.01 mM for Pb2+. Much higher levels of Pb were required to inhibit [14C]glucose mineralization directly in soils. In microcosm experiments with these samples, microbial biomass and the ratio of microbial biomass to soil organic C were not correlated with the concentrations of hydrocarbons and heavy metals. However, microbial C respiration in samples with a higher level of hydrocarbons differed from the other soils no matter whether complex organic C (alfalfa) was added or not. The ratios of microbial C respiration to microbial biomass differed significantly among the soil samples (P < 0.05) and were relatively high in soils contaminated with hydrocarbons or heavy metals. Our results suggest that the soil microbial community was predominantly affected by hydrocarbons.     

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.     

川西亚高山不同林龄云杉人工林土壤微生物群落结构

以川西亚高山云杉人工林林地土壤为对象,采用磷脂脂肪酸(PLFA)法研究了4种不同林龄(50、38、27和20年)的人工林土壤微生物多样性和群落结构特征.结果表明: 随着林龄的增加,土壤有机碳和全氮含量逐步增加;土壤微生物Shannon多样性和Pielou均匀度指数则呈现先增后减的趋势.土壤微生物总PLFAs量、细菌PLFAs量、真菌PLFAs量、放线菌PLFAs量以及丛枝菌根真菌PLFAs量均表现为随林龄的增加而增加.主成分分析(PCA)表明,不同林龄人工林的土壤微生物群落结构之间存在显著差异,其中,第1主成分(PC1)和第2主成分(PC2)共同解释了土壤微生物群落结构总变异的66.8%.冗余分析(RDA)表明,对土壤微生物群落结构产生显著影响的环境因子分别为土壤有机碳、全氮、全钾以及细根生物量.随着人工造林时间的延长,土壤肥力和微生物生物量增加,森林生态系统的恢复进程稳定.     

石油污染土壤水改旱田后污染物组份及微生物群落结构变化

对沈抚灌区水改旱田不同年限土壤的石油污染物浓度及组分进行了分析,并采用变性梯度凝胶电泳和磷脂脂肪酸分析方法,分析了污染土壤微生物群落结构的变化.结果表明：1)石油污染土壤水改旱田后,年限越长,总多环芳烃在总石油烃中所占的比重越大,高分子量多环芳烃在总多环芳烃中所占比重也越大;2)总磷脂脂肪酸量与总石油烃呈显著正相关,与总多环芳烃相关性不显著;3)两种方法对土壤微生物群落结构的分析得出的结论一致,石油污染土壤微生物群落结构主要与其相对地理位置有关,当污染物的浓度达到一定程度时,土壤微生物群落结构会发生明显的改变.     

Microbial Biomass and Community Structure in a Sequence of Soils with Increasing Fertility and Changing Land Use

Abstract The microbial biomass and community structure of eight Chinese red soils with different fertility and land use history was investigated. Two community based microbiological measurements, namely, community level physiological profiling (CLPP) using Biolog sole C source utilization tests and phospholipid fatty acid (PLFA) profiles, were used to investigate the microbial ecology of these soils and to determine how land use alters microbial community structure. Microbial biomass-C and total PLFAs were closely correlated to organic carbon and total nitrogen, indicating that these soil microbial measures are potentially good indices of soil fertility in these highly weathered soils. Metabolic quotients and C source utilization were not correlated with organic carbon or microbial biomass. Multivariate analysis of sole carbon source utilization patterns and PLFAs demonstrated that land use history and plant cover type had a significant impact on microbial community structure. PLFAs showed these differences more than CLPP methods. Consequently, PLFA analysis was a better method for assessing broad-spectrum community differences and at the same time attempting to correlate changes with soil fertility. Soils from tea orchards were particularly distinctive in their CLPP. A modified CLPP method, using absorbance readings at 405 nm and different culture media at pH values of 4.7 and 7.0, showed that the discrimination obtained can be influenced by the culture conditions. This method was used to show that the distinctive microbial community structure in tea orchard soils was not, however, due to differences in pH alone. Received: 1 December 1999; Accepted: 6 June 2000; Online Publication: 28 August 2000     

Estimating high-affinity methanotrophic bacterial biomass, growth, and turnover in soil by phospholipid fatty acid 13C labeling

A time series phospholipid fatty acid (PLFA) 13C-labeling study was undertaken to determine methanotrophic taxon, calculate methanotrophic biomass, and assess carbon recycling in an upland brown earth soil from Bronydd Mawr (Wales, United Kingdom). Laboratory incubations of soils were performed at ambient CH4 concentrations using synthetic air containing 2 parts per million of volume of 13CH4. Flowthrough chambers maintained a stable CH4 concentration throughout the 11-week incubation. Soils were analyzed at weekly intervals by gas chromatography (GC), GC-mass spectrometry, and GC-combustion-isotope ratio mass spectrometry to identify and quantify individual PLFAs and trace the incorporation of 13C label into the microbial biomass. Incorporation of the 13C label was seen throughout the experiment, with the rate of incorporation decreasing after 9 weeks. The delta13C values of individual PLFAs showed that 13C label was incorporated into different components to various extents and at various rates, reflecting the diversity of PLFA sources. Quantitative assessments of 13C-labeled PLFAs showed that the methanotrophic population was of constant structure throughout the experiment. The dominant 13C-labeled PLFA was 18:1omega7c, with 16:1omega5 present at lower abundance, suggesting the presence of novel type II methanotrophs. The biomass of methane-oxidizing bacteria at optimum labeling was estimated to be about 7.2 x 10(6) cells g(-1) of soil (dry weight). While recycling of 13C label from the methanotrophic biomass must occur, it is a slower process than initial 13CH4 incorporation, with only about 5 to 10% of 13C-labeled PLFAs reflecting this process. Thus, 13C-labeled PLFA distributions determined at any time point during 13CH4 incubation can be used for chemotaxonomic assessments, although extended incubations are required to achieve optimum 13C labeling for methanotrophic biomass determinations.     

Geographic Variation in Plant Community Structure of Salt Marshes: Species,Functional and Phylogenetic Perspectives

In general, community similarity is thought to decay with distance; however, this view may be complicated by the relative roles of different ecological processes at different geographical scales, and by the compositional perspective (e.g. species, functional group and phylogenetic lineage) used. Coastal salt marshes are widely distributed worldwide, but no studies have explicitly examined variation in salt marsh plant community composition across geographical scales, and from species, functional and phylogenetic perspectives. Based on studies in other ecosystems, we hypothesized that, in coastal salt marshes, community turnover would be more rapid at local versus larger geographical scales; and that community turnover patterns would diverge among compositional perspectives, with a greater distance decay at the species level than at the functional or phylogenetic levels. We tested these hypotheses in salt marshes of two regions: The southern Atlantic and Gulf Coasts of the United States. We examined the characteristics of plant community composition at each salt marsh site, how community similarity decayed with distance within individual salt marshes versus among sites in each region, and how community similarity differed among regions, using species, functional and phylogenetic perspectives. We found that results from the three compositional perspectives generally showed similar patterns: there was strong variation in community composition within individual salt marsh sites across elevation; in contrast, community similarity decayed with distance four to five orders of magnitude more slowly across sites within each region. Overall, community dissimilarity of salt marshes was lowest on the southern Atlantic Coast, intermediate on the Gulf Coast, and highest between the two regions. Our results indicated that local gradients are relatively more important than regional processes in structuring coastal salt marsh communities. Our results also suggested that in ecosystems with low species diversity, functional and phylogenetic approaches may not provide additional insight over a species-based approach.