Stability in Natural Bacterial Communities: II. Plant Resource Allocation Effects on Rhizosphere Diazotroph Assemblage Composition

Abstract Plant rhizospheres are dynamic environments in which microbes compete for resources, particularly plant-derived organic carbon/energy sources. Altering plant productivity changes the availability of carbon/energy resources to rhizosphere microbiota. This limitation is expected to intensify competition for the remaining carbon supply and could cause the loss of poor competitors from the assemblage. We clipped or shaded plots of the salt marsh cord grass, Spartina alterniflora, to shift plant carbon resource allocation from the rhizosphere to the aboveground shoots. We then examined key porewater parameters (pH, salinity, H2S, NH⁺4), diazotrophic activity (by acetylene reduction assay), and diazotroph assemblage composition after 2 weeks or 8 weeks of treatment. The diazotroph assemblage composition was monitored via the polymerase chain reaction using nifH specific primers followed by denaturing gradient gel electrophoresis (DGGE) analysis. Porewater parameters and acetylene reduction rates did not differ significantly among treatments. The DGGE profiles also were very similar across the control and experimental treatments, indicating that no detectable diazotroph species were displaced from the assemblage. This implies that rhizosphere diazotrophs are able to compete successfully against nondiazotrophs, in spite of the high energy requirements of nitrogen fixation. These results show that the species composition of the diazotroph assemblage in the S. alterniflora rhizosphere is stable in the face of short-duration but potentially high-impact variations in carbon resource availability.     

Spatial and Temporal Assessment of Diazotroph Assemblage Composition in Vegetated Salt Marsh Sediments Using Denaturing Gradient Gel Electrophoresis Analysis

Abstract Diazotroph assemblage compositions were assessed in rhizosphere sediments from the tall and short form Spartina alterniflora growth zones over an annual cycle. Sediment cores were collected for DNA extraction and nitrogenase (acetylene reduction) activity assays, and porewater samples were analyzed for several chemical parameters in March, June, September, and December 1997. These data were collected to determine if within- or between-zone differences in the diazotroph assemblage composition correlated with differences in key environmental variables or acetylene reduction activity. Acetylene reduction rates differed between zones and within a zone over an annual period. Soluble sulfide concentrations were higher in the short form S. alterniflora zone on all dates except those in June and differed within both zones on different sample dates. nifH sequences were recovered from rhizosphere sediment DNA by PCR amplification using nifH specific primers. These amplimers were analyzed using denaturing gradient gel electrophoresis (DGGE), and the resulting patterns were compared by neural network and linear discriminant analyses. Ten prominent amplimers, four of which were apparent heteroduplexes, were observed. DGGE banding profiles showed minor differences among sampling dates and between sample zones, but the overall banding pattern was remarkably consistent. This reflects overall similarity between the amplifiable diazotroph assemblages in the tall and short S. alterniflora growth zones and substantial seasonal stability in assemblage composition. Received: 2 March 1999; Accepted: 4 May 1999     

Nutrient cycling relative to δ<Superscript>15</Superscript>N and δ<Superscript>13</Superscript>C natural abundance in a coastal wetland with long-term nutrient additions

Because nitrogen and phosphorus are primary resources for plant, algal, and microbial production, increases in nutrient inputs can markedly alter aquatic ecosystems. Coastal wetland plots at Belle W. Baruch Marine Field Laboratory (South Carolina, USA) have been amended with nitrogen and phosphorus for ~20 years to determine the effects of nutrient loading on coastal wetlands. We conducted a survey of δ¹⁵N and δ¹³C natural abundance in coastal wetland organic pools (sediment, vegetation) with long-term nutrient amendments (control, no addition; nitrogen; phosphorus; and nitrogen + phosphorus additions). Additionally, we conducted laboratory assays to quantify pore water nutrient availability and nitrification rates. Marsh vegetation (Spartina alterniflora) had enriched δ¹³C values (mean −14‰) relative to bulk sediment samples (mean −18‰). Nitrogen-amended plots (alone and in combination with phosphorus) had enriched δ¹³C values in the surface sediment (0–5 cm; mean −16.1‰) relative to control (mean −16.5‰) and phosphorus-amended plots (mean −16.8‰). Nitrogen-amended plots also had depleted δ¹⁵N in S. alterniflora leaf tissues (−3.3‰) and surface sediment samples (mean 2.1‰) relative to leaf tissues (mean 2.1‰) or sediment samples (mean 5.8‰) from control or phosphorus-only amended plots. Nitrate availability (as increased pore water concentration) was higher in N-amended plots although ammonium availability did not differ. Phosphorus availability was higher only in phosphorus-only amended plots. Overall, we found that long-term nutrient amendments to coastal wetlands significantly altered nutrient availability and uptake rates as well as natural abundance of δ¹³C and δ¹⁵N in multiple organic matter sources.     

Carbon-Degrading Enzyme Activities Stimulated by Increased Nutrient Availability in Arctic Tundra Soils

Climate-induced warming of the Arctic tundra is expected to increase nutrient availability to soil microbes, which in turn may accelerate soil organic matter (SOM) decomposition. We increased nutrient availability via fertilization to investigate the microbial response via soil enzyme activities. Specifically, we measured potential activities of seven enzymes at four temperatures in three soil profiles (organic, organic/mineral interface, and mineral) from untreated native soils and from soils which had been fertilized with nitrogen (N) and phosphorus (P) since 1989 (23 years) and 2006 (six years). Fertilized plots within the 1989 site received annual additions of 10 g N⋅m^-2⋅year^-1 and 5 g P⋅m^-2⋅year^-1. Within the 2006 site, two fertilizer regimes were established – one in which plots received 5 g N⋅m^-2⋅year^-1 and 2.5 g P⋅m^-2⋅year^-1 and one in which plots received 10 g N⋅m^-2⋅year^-1 and 5 g P⋅m^-2⋅year^-1. The fertilization treatments increased activities of enzymes hydrolyzing carbon (C)-rich compounds but decreased phosphatase activities, especially in the organic soils. Activities of two enzymes that degrade N-rich compounds were not affected by the fertilization treatments. The fertilization treatments increased ratios of enzyme activities degrading C-rich compounds to those for N-rich compounds or phosphate, which could lead to changes in SOM chemistry over the long term and to losses of soil C. Accelerated SOM decomposition caused by increased nutrient availability could significantly offset predicted increased C fixation via stimulated net primary productivity in Arctic tundra ecosystems.     

Molecular Analysis of Diazotroph Diversity in the Rhizosphere of the Smooth Cordgrass, Spartina alterniflora

N₂ fixation by diazotrophic bacteria associated with the roots of the smooth cordgrass, Spartina alterniflora, is an important source of new nitrogen in many salt marsh ecosystems. However, the diversity and phylogenetic affiliations of these rhizosphere diazotrophs are unknown. Denaturing gradient gel electrophoresis (DGGE) of PCR-amplified nifH sequence segments was used in previous studies to examine the stability and dynamics of the Spartina rhizosphere diazotroph assemblages in the North Inlet salt marsh, near Georgetown, S.C. In this study, plugs were taken from gel bands from representative DGGE gels, the nifH amplimers were recovered and cloned, and their sequences were determined. A total of 59 sequences were recovered, and the amino acid sequences predicted from them were aligned with sequences from known and unknown diazotrophs in order to determine the types of organisms present in the Spartina rhizosphere. We recovered numerous sequences from diazotrophs in the γ subdivision of the division Proteobacteria (γ-Proteobacteria) and from various anaerobic diazotrophs. Diazotrophs in the α-Proteobacteria were poorly represented. None of the Spartina rhizosphere DGGE band sequences were identical to any known or previously recovered environmental nifH sequences. The Spartina rhizosphere diazotroph assemblage is very diverse and apparently consists mainly of unknown organisms.     

Seasonal Variability of Diazotroph Assemblages Associated with the Rhizosphere of the Salt Marsh Cordgrass, Spartina alterniflora

Nitrogen fixation is the primary N source in the highly productive but N-limited North Inlet, SC, USA salt marsh system. The diverse assemblages of nitrogen-fixing (diazotrophic) bacteria associated with the rhizospheres of the short and tall growth forms of Spartina alterniflora were analyzed at two sites, Crab Haul Creek and Goat Island, which are in different tidal creek drainage systems in this marsh. The sites differed in proximity to the main channel for tidal intrusion and in several edaphic parameters. We hypothesized that either the differing abiotic environmental regimes of the two sites or the variation due to seasonal effects result in differences in the diazotroph assemblage. Rhizosphere samples were collected seasonally during 1999 and 2000. DNA was purified and nifH amplified for denaturing gradient gel electrophoresis (DGGE) analysis of diazotroph assemblage composition. Principal components analysis was used to analyze the binary DGGE band position data. Season strongly influenced assemblage composition and biplots were used to identify bands that significantly affected the seasonal and site-specific clustering. The types of organisms that were most responsive to seasonal or site variability were identified on the basis of DGGE band sequences. Seasonally responsive members of the anaerobic diazotrophs were detected during the winter and postsenescence conditions and may have been responsible for elevated pore water sulfide concentrations. Sequences from a diverse assemblage of Gammaproteobacteria were predominant during growth periods of S. alterniflora. Abiotic environmental parameters strongly influenced both the S. alterniflora and the diazotrophic bacterial assemblages associated with this keystone salt marsh plant species.     

Diel plant water use and competitive soil cation exchange interact to enhance NH<Subscript>4</Subscript><Superscript>+</Superscript> and K<Superscript>+</Superscript> availability in the rhizosphere

Aims

Hydro-biogeochemical processes in the rhizosphere regulate nutrient and water availability, and thus ecosystem productivity. We hypothesized that two such processes often neglected in rhizosphere models — diel plant water use and competitive cation exchange — could interact to enhance availability of K⁺ and NH₄ ⁺, both high-demand nutrients.

Methods

A rhizosphere model with competitive cation exchange was used to investigate how diel plant water use (i.e., daytime transpiration coupled with no nighttime water use, with nighttime root water release, and with nighttime transpiration) affects competitive ion interactions and availability of K⁺ and NH₄ ⁺.

Results

Competitive cation exchange enabled low-demand cations that accumulate against roots (Ca²⁺, Mg²⁺, Na⁺) to desorb NH₄ ⁺ and K⁺ from soil, generating non-monotonic dissolved concentration profiles (i.e. ‘hotspots’ 0.1–1 cm from the root). Cation accumulation and competitive desorption increased with net root water uptake. Daytime transpiration rate controlled diel variation in NH₄ ⁺ and K⁺ aqueous mass, nighttime water use controlled spatial locations of ‘hotspots’, and day-to-night differences in water use controlled diel differences in ‘hotspot’ concentrations.

Conclusions

Diel plant water use and competitive cation exchange enhanced NH₄ ⁺ and K⁺ availability and influenced rhizosphere concentration dynamics. Demonstrated responses have implications for understanding rhizosphere nutrient cycling and plant nutrient uptake.

     

Responses of Salt Marsh Plant Rhizosphere Diazotroph Assemblages to Changes in Marsh Elevation,Edaphic Conditions and Plant Host Species

An important source of new nitrogen in salt marsh ecosystems is microbial diazotrophy (nitrogen fixation). The diazotroph assemblages associated with the rhizospheres (sediment directly affected by the roots) of salt marsh plants are highly diverse, somewhat stable, and consist mainly of novel organisms. In Crab Haul Creek Basin, North Inlet, SC, the distribution of plant types into discrete zones is dictated by relatively minor differences in marsh elevation and it was hypothesized that the biotic and abiotic properties of the plant zones would also dictate the composition of the rhizosphere diazotroph assemblages. Over a period of 1 year, rhizosphere sediments were collected from monotypic stands of the black needlerush, Juncus roemerianus, the common pickleweed, Salicornia virginica, the short and tall growth forms of the smooth cordgrass Spartina alterniflora, and a mixed zone of co-occurring S. virginica and short form, S. alterniflora. DNA was extracted, purified and nifH sequences PCR amplified for denaturing gradient gel electrophoresis (DGGE) analysis to determine the composition of the diazotroph assemblages. The diazotroph assemblages were strongly influenced by season, abiotic environmental parameters and plant host. Sediment chemistry and nitrogen fixation activity were also significantly influenced by seasonal changes. DGGE bands that significantly affected seasonal and zone specific clustering were identified and most of these sequences were from novel diazotrophs, unaffiliated with any previously described organisms. At least one third of the recovered nifH sequences were from a diverse assemblage of Chlorobia, and γ-, α-, β- and δ-Proteobacteria. Diazotrophs that occurred throughout the growing season and among all zones (frequently detected) were also mostly novel. These significant sequences indicated that diazotrophs driving the structure of the assemblages were diverse, versatile, and some were ubiquitous while others were seasonally responsive. Several ubiquitous sequences were closely related to sequences of actively N₂ fixing diazotrophs previously recovered from this system. These sequences from ubiquitous and versatile organisms likely indicate the diazotrophs in these rhizosphere assemblages that significantly contribute to ecosystem function.     

Molecular analysis of diazotroph diversity in the rhizosphere of the smooth cordgrass, Spartina alterniflora

N(2) fixation by diazotrophic bacteria associated with the roots of the smooth cordgrass, Spartina alterniflora, is an important source of new nitrogen in many salt marsh ecosystems. However, the diversity and phylogenetic affiliations of these rhizosphere diazotrophs are unknown. Denaturing gradient gel electrophoresis (DGGE) of PCR-amplified nifH sequence segments was used in previous studies to examine the stability and dynamics of the Spartina rhizosphere diazotroph assemblages in the North Inlet salt marsh, near Georgetown, S.C. In this study, plugs were taken from gel bands from representative DGGE gels, the nifH amplimers were recovered and cloned, and their sequences were determined. A total of 59 sequences were recovered, and the amino acid sequences predicted from them were aligned with sequences from known and unknown diazotrophs in order to determine the types of organisms present in the Spartina rhizosphere. We recovered numerous sequences from diazotrophs in the gamma subdivision of the division Proteobacteria (gamma-Proteobacteria) and from various anaerobic diazotrophs. Diazotrophs in the alpha-Proteobacteria were poorly represented. None of the Spartina rhizosphere DGGE band sequences were identical to any known or previously recovered environmental nifH sequences. The Spartina rhizosphere diazotroph assemblage is very diverse and apparently consists mainly of unknown organisms.     

Nitrogen mineralization in a green manure-amended soil as influenced by cropping history and subsequent crop

A greenhouse experiment was conducted to determine the influence of cropping variables on nitrogen dynamics in a soil amended with green manure. Surface soil from various long-term spring wheat rotations was amended with¹⁵N-labelled legume green manure (Lathyrus tingitanus) and subsequently cropped (canola [Brassica napus] and spring wheat [Triticum aestivum]) or incubated without a crop for 56 days in a greenhouse. Nitrogen mineralization from both the indigenous soil N and from green manure was suppressed in cropped soil. Net N mineralization in the uncropped and cropped treatments averaged 73 and 43 mg kg⁻¹, respectively. This difference was attributed, in part, to enhanced biological immobilization in the rhizosphere. Previous cropping practices also had significant effect on N mineralization, largely by their influence on indigenous organic matter quality. These observations suggest that short-term N mineralization is favored by fallowing soil after green manure application whereas N retention in organic matter is favored by immediate cropping. Contribution 3878873     

Variation in Microbial Community Composition and Culturability in the Rhizosphere of Leucanthemopsis alpina (L.) Heywood and Adjacent Bare Soil Along an Alpine Chronosequence

We compared the size, culturability, diversity, and dominant species similarity of the bacterial communities of Leucanthemopsis alpina (L.) Heywood rhizosphere and adjacent bare soil (interspace) along a chronosequence of soil development time (5, 50, and 70 years) in the forefield of the Dammaglacier (Switzerland). We found no evidence that the size of the bacterial community was significantly affected by either soil age or the presence of L. alpina. In contrast, the proportion of the bacterial community that could be cultured on nonselective agars, and which was taken as an indication of the proportion of r-selected populations, was significantly higher in the 50- and 70-year-old soils than in the 5-year-old soil, and was also significantly higher in the rhizosphere of L. alpina at all time points. RDA indicated significant correlations between the increased culturability of the bacterial community over time and increasing concentrations of labile N, and between the increased culturability in the rhizosphere and increased concentrations of labile C and N. HaeIII-amplified ribosomal DNA (rDNA) restriction analysis of a library of 120 clones of 16S rDNA revealed 85 distinct phylotypes. Hurlbert's probability of interspecific encounter (PIE) values derived from this library ranged from 0.95 to 1.0, indicating a very high genetic diversity. There was no significant difference in the PIE values of rhizosphere and interspace communities. Detrended correspondence analysis (DCA) of 16S ribosomal RNA (rRNA) denaturing gradient gel electrophoresis (DGGE) community profiles clearly distinguished the rhizosphere from the interspace community in the 5-year-old soils and also clearly distinguished between these communities and the rhizosphere and interspace communities of the 50- and 70-year-old soils. However, 16S rRNA DGGE revealed little difference between rhizosphere and interspace communities in the 50- and 70-year-old soils. The relative similarity of the 16S rRNA profiles strongly reflected labile carbon and nitrogen availability. Overall, our results suggest that improved C and N availability in the rhizosphere of L. alpina increases the size of r-selected bacterial species populations, but that the influence of L. alpina depends on soil age, being maximal in the youngest soils and minimal in the oldest. The reduced influence of L. alpina in the older soils may reflect a feedback between improved nutrient availability and reduced rhizodeposition.     

施用污泥对油菜根际养分和不同种类重金属的影响

根际是控制植物养分动态的重要因素,养分动态也影响着根际土壤环境。当土壤被污水污泥改良后,根际土壤中的养分和重金属性质也会发生变化。目前很少有人研究施用污泥的土壤中植物根系对根际重金属有效性和分布的影响。采用根垫—冰冻薄层切片法对施用污泥后土壤中油菜根际的养分和重金属分布情况进行研究,以期探明污泥改良土壤中根际重金属的活化特征。当土壤施用污泥后,根际土壤中DTPA提取态Zn,Cd,Ni,Mn,有效磷,有效钾和铵态氮被显著消耗,而根际土壤中DTPA提取态Cu没有明显的消耗或积累。当土壤中施用大量污泥时,根际土壤的pH值随着离根表面距离的增加而增加。无论土壤是否用污泥处理,油菜根际土壤中可交换态Cu都显著减少。当土壤被50%污泥改良时,在距离根表面0—2 mm处的油菜根际土壤中碳酸盐结合态,铁锰氧化物结合态,有机物结合态,残渣态的Cu和Zn都被消耗较多。污泥的施用对油菜的生长有促进作用。随着污泥施用量的增加,油菜地上部分Cu和Zn的含量没有显著变化。施用污泥量小于25%的土壤中,污泥没有增加重金属的可利用性和移动性。除了Cu,油菜根际土壤中DTPA提取态Zn,Cd,Ni的减少表明施用污泥的土壤中重金属的活化是非常有限的。     

荒漠草原不同放牧强度背景下添加氮水对凋落物分解的影响

凋落物分解是连接生态系统地上、地下过程的重要环节,决定了生态系统养分循环速率,但到目前为止对凋落物分解在荒漠草地生态系统受放牧以及外源资源补给影响的研究较少。本研究通过对不同放牧强度(对照、轻牧、中牧和重牧)短花针茅草原群落进行添加氮素(10.0 g N m~(-2) a~(-1))和增水(108 mm/a)处理,探讨群落水平凋落物分解速率的变化。研究结果显示,过去不同强度放牧历史对群落凋落物分解影响极显著(P0.0001)。凋落物前期分解(135 d)过程中,凋落物初始C∶N比与凋落物分解速率常数呈显著负相关关系,表明凋落物可降解性在凋落物前期分解中起主要作用。轻度放牧影响下凋落物分解速度最快,这与该条件下凋落物C∶N比显著低于其他放牧强度下的有关,说明适度放牧不仅有利于群落维持,也在一定程度上有利于生态系统养分循环。当凋落物分解更长时间(870 d)后,对照区凋落物分解速率显著低于放牧处理样地,但凋落物初始C∶N比对凋落物分解速率没有显著影响。进一步分析显示,不同放牧强度背景下长期凋落物分解速率与分解环境的土壤微生物多样性成正相关关系,与群落盖度呈极显著(P0.001)负相关关系。添加氮素显著(P0.05)降低凋落物分解速度,但对凋落物氮含量无显著影响。生长季加水未影响凋落物质量及凋落物分解速度。研究结果表明,凋落物前期分解受凋落物质量影响,但较长时间凋落物分解则与分解过程中接受到的太阳辐射量有关。     

Taxonomic patterns in the nitrogen assimilation of soil prokaryotes

Nitrogen (N) is frequently a limiting nutrient in soil; its availability can govern ecosystem functions such as primary production and decomposition. Assimilation of N by microorganisms impacts the availability of N in soil. Despite its established ecological significance, the contributions of microbial taxa to N assimilation are unknown. Here we measure N uptake and use by microbial phylotypes and taxonomic groups within a diverse assemblage of soil microbes through quantitative stable isotope probing (qSIP) with ¹⁵N. Following incubation with ¹⁵ , distinct patterns of ¹⁵N assimilation among taxonomic groups were observed. For instance, glucose addition stimulated ¹⁵N assimilation in most members of Actinobacteria and Proteobacteria but generally decreased ¹⁵N use by Firmicutes and Bacteriodetes. While is considered a preferred and universal source of N to prokaryotes, the majority (> 80%) of N assimilation in our soils could be attributed to a handful of active orders. Characterizing N assimilation of taxonomic groups with ¹⁵N qSIP may provide a basis for understanding how microbial community composition influences N availability in the environment.     

Heat stress reduces the contribution of diazotrophs to coral holobiont nitrogen cycling

Efficient nutrient cycling in the coral-algal symbiosis requires constant but limited nitrogen availability. Coral-associated diazotrophs, i.e., prokaryotes capable of fixing dinitrogen, may thus support productivity in a stable coral-algal symbiosis but could contribute to its breakdown when overstimulated. However, the effects of environmental conditions on diazotroph communities and their interaction with other members of the coral holobiont remain poorly understood. Here we assessed the effects of heat stress on diazotroph diversity and their contribution to holobiont nutrient cycling in the reef-building coral Stylophora pistillata from the central Red Sea. In a stable symbiotic state, we found that nitrogen fixation by coral-associated diazotrophs constitutes a source of nitrogen to the algal symbionts. Heat stress caused an increase in nitrogen fixation concomitant with a change in diazotroph communities. Yet, this additional fixed nitrogen was not assimilated by the coral tissue or the algal symbionts. We conclude that although diazotrophs may support coral holobiont functioning under low nitrogen availability, altered nutrient cycling during heat stress abates the dependence of the coral host and its algal symbionts on diazotroph-derived nitrogen. Consequently, the role of nitrogen fixation in the coral holobiont is strongly dependent on its nutritional status and varies dynamically with environmental conditions.Subject terms: Microbial ecology, Climate-change ecology     

Analysis of the endophytic lifestyle and plant growth promotion of Burkholderia terricola ZR2-12

Nitrogen (N) enrichment has been pinpointed as a main driver for biodiversity change. Most of our knowledge of effects of increased N availability on ecosystems comes from northern Europe and America. Most other ecosystem types have been neglected. In contribution to filling this gap, our study examined the short-term effects of N enrichment in a N-manipulation (doses and forms) field study of a severely nutrient-limited Mediterranean ecosystem located in a Natura 2000 site in Portugal. Our aims were to (a) understand the effects of N enrichment on plant diversity, and to (b) link N-driven plant community changes with changes in soil inorganic N availability. In general, the standing plant community responded to short-term N enrichment with increased richness and evenness. Changes in the plant community occurred through changes in species composition and cover, and were correlated with soil N, and N and phosphorus availability. Fertilization with 80 kg NH₄NO₃ ha^?1 y^?1 was the treatment which changed plant composition the most, while geophytes, hemicryptophytes and therophytes were the biological types more responsive to N enrichment. Dittrichia viscosa was the only species that responded significantly to increased N, i.e., its cover decreased in control plots, but increased in fertilized plots, suggesting that it could be used as an indicator of N enrichment in Mediterranean maquis. Changes in plant richness and evenness were correlated with the mean and/or the variation (standard deviation) of soil inorganic N parameters (e.g. nitrate concentration in the soil solution and the soil??s ratio of bioavailable N and phosphorus) measured along the time between the two plant community assessments. However, short- and long-term effects can be quite distinct, thus highlighting the need for further studies.     

Does nitrogen availability control rates of litter decomposition in forests?

The effects of increased exogenous N availability on rates of litter decomposition were assessed in several field fertilization trials. In a jack pine (Pinus banksiana Lamb.) forest, needle litter decomposed at the same rate in control plots and in plots fertilized with urea and ammonium nitrate (1350 kg N ha^-1) with or without P and K. Mixed needle litter of western hemlock (Tsuga heterophylla (Raf.) Sarg.), western red cedar (Thuja plicata Donn) and Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) incubated in plots recently amended with sewage sludge (500 kg N ha^-1) lost less weight during 3 years than did litter in control plots. Forest floor material also decomposed more slowly in plots amended with sewage sludge. Paper birch (Betula papyrifera Marsh.) leaf litter placed on sewage sludge (1000 kg N ha^-1), pulp sludge, or sewage-pulp sludge mixtures decomposed at the same rate as leaf litter in control plots. These experiments demonstrate little effect of exogenous N availability on rates of litter decomposition.The influence of endogenous N availability on rates of litter decomposition was examined in a microcosm experiment. Lodgepole pine (Pinus contorta var. latifolia Engelm.) needle litter collected from N-fertilized trees (525 kg N ha^-1 in ammonium nitrate) were 5 times richer in N than needles from control trees (1.56% N versus 0.33% N in control trees), but decomposed at the same rate. Green needles from fertilized trees contained twice as much N as needles from control trees (1.91% N versus 0.88% N), but decomposed at the same rate. These experiments suggest that N availability alone, either exogenous or endogenous, does not control rates of litter decomposition. Increased N availability, through fertilization or deposition, in the absence of changes in vegetation composition, will not alter rates of litter decomposition in forests.     

Field response of rice paddy crop to Azospirillum inoculation: physiology of rhizosphere bacterial communities and the genetic diversity of endophytic bacteria in different parts of the plants

The response of rice plants to the application of inoculant containing two Azospirillum brasilense strains was studied under field conditions. The experiment was performed as three treatments with four replicates in randomized complete blocks arranged as plots of 60 m² in an area on a Vertic Argiudol soil type in the province of Entre Ríos, Argentina. The bacterial rhizosphere community and also the diazotrophic isolates obtained from control and inoculated rice plants were analyzed in relation to their physiology and biological nitrogen fixation (BNF). The MPN of diazotrophs in the rhizosphere varied during the ontogenic cycle. The patterns of distribution of the microbial physiological activities obtained by principal component analysis of community-level physiological profiles (CLPP) showed differences in the utilization of carbon sources by the rhizosphere communities among treatments. Although the analyses of DGGE 16S and nifH profiles have not indicated that the inoculation influenced the genetic diversity of bacterial communities among treatments, they revealed that the banding profiles were altered in different parts of the rice plant by each Azospirillum inoculation treatment. These observations suggest that physiological responses of plant tissues to the inoculation may have occurred. According to agronomic parameters of each treatment, the Azospirillum inoculation increased aerial biomass at the tillering and grain-filling stages. Although the N content accumulated in rice plants increased by 16 and 50 kg ha^?1, the BNF contribution could not be estimated under our experimental conditions by the ¹⁵N balance technique. Based on this field inoculation experiment to rice plants, it is noteworthy that our data suggest that due to Azospirillum inoculation the increase of total N accumulated in rice plants could be a tool to help farmers to improve production and maintain high input of plant residues, providing more organic matter to the soil and guaranteeing sustainability of the system.     

樟树人工林凋落物养分含量及归还量对氮沉降的响应

氮沉降的持续增加对陆地生态系统的健康发展构成严重威胁,森林是陆地生态系统中重要的组成部分,大量的氮沉降对其结构和功能造成严重影响。凋落物是森林生态系统养分循环的重要组成部分,它对土壤肥力、森林生态系统养分循环等方面具有重要作用。为了探讨亚热带常绿阔叶森林凋落物对氮沉降增加的响应,在湖南省森林植物园以樟树人工林为研究对象进行模拟氮沉降的实验,实验设置4种氮添加水平CK(0g N m~(-2)a~(-1),对照)、LN(5g N m~(-2)a~(-1)),MN(15g N m~(-2)a~(-1)),HN(30g N m~(-2)a~(-1)),研究氮沉降对樟树林年凋落物量、凋落物养分含量以及归还量的影响。结果表明:不同施氮水平下(CK、LN、MN、HN),樟树林凋落物的年凋落量分别为(4.53±0.32)t hm~(-2)a~(-1)、(3.95±0.28)t hm~(-2)a~(-1)、(3.56±0.41)t hm~(-2)a~(-1)、(4.46±0.48)t hm~(-2)a~(-1),施氮抑制了樟树林的凋落量,且低、中氮处理下差异显著(P0.05);施氮处理后凋落物的养分含量大小顺序为:CNCaKMg,凋落物的碳含量没有显著变化,但氮含量都有所增加,因此,施氮降低了樟树凋落物各组分的C/N比;凋落物中元素的年归还量大小顺序表现为:CNCaKMg,施氮处理对凋落物C、K、Ca、Mg归还量有抑制作用,但对凋落物N归还量表现为促进作用。     

Long-term effects of experimental nitrogen additions on foliar litter decay and humus formation in forest ecosystems

Decomposition rates and N dynamics of foliar litter from 4 tree species were measured over a 72 month period on the Chronic Nitrogen Addition plots at the Harvard Forest, Petersham MA, beginning in November 1988. Plots received nitrogen additions of 0, 5 and 15 g N m^-2yr^-1 in two different stand types: red pine and mixed hardwood. Bags were collected in August and November of each year and litter analysed for mass remaining, nitrogen, cellulose and lignin content. Mass remaining was significantly greater for litter in nitrogen treated plots than in control plots after 48 months. Lignin content of litter was significantly higher with nitrogen treatments but there was little effect of treatment on cellulose content. N concentration was similar between treatments, but greater mass remaining in treated plots resulted in a higher total amount of N in humus produced in the high N plot. This mechanism could be a sink for up to 1.5 g N·m^-2yr^-1 of the 1.5 g N·m^-2yr^-1 added annually to the high N plots. Reduced decomposition rates in conjunction with increased lignin accumulation could impact global carbon sequestration as well.