Microbial responses to nitrogen addition in three contrasting grassland ecosystems

The effects of global N enrichment on soil processes in grassland ecosystems have received relatively little study. We assessed microbial community response to experimental increases in N availability by measuring extracellular enzyme activity (EEA) in soils from three grasslands with contrasting edaphic and climatic characteristics: a semiarid grassland at the Sevilleta National Wildlife Refuge, New Mexico, USA (SEV), and mesic grasslands at Konza Prairie, Kansas, USA (KNZ) and Ukulinga Research Farm, KwaZulu-Natal, South Africa (SAF). We hypothesized that, with N enrichment, soil microbial communities would increase C and P acquisition activity, decrease N acquisition activity, and reduce oxidative enzyme production (leading to recalcitrant soil organic matter [SOM] accumulation), and that the magnitude of response would decrease with soil age (due to higher stabilization of enzyme pools and P limitation of response). Cellulolytic activities followed the pattern predicted, increasing 35–52% in the youngest soil (SEV), 10–14% in the intermediate soil (KNZ) and remaining constant in the oldest soil (SAF). The magnitude of phosphatase response did not vary among sites. N acquisition activity response was driven by the enzyme closest to its pH optimum in each soil: i.e., leucine aminopeptidase in alkaline soil, β-N-acetylglucosaminidase in acidic soil. Oxidative enzyme activity varied widely across ecosystems, but did not decrease with N amendment at any site. Likewise, SOM and %C pools did not respond to N enrichment. Between-site variation in both soil properties and EEA exceeded any treatment response, and a large portion of EEA variability (leucine aminopeptidase and oxidative enzymes), 68% as shown by principal components analysis, was strongly related to soil pH (r = 0.91, P < 0.001). In these grassland ecosystems, soil microbial responses appear constrained by a molecular-scale (pH) edaphic factor, making potential breakdown rates of SOM resistant to N enrichment.     

Microbial biomass C,N and P in two arctic soils and responses to addition of NPK fertilizer and sugar: implications for plant nutrient uptake

The soil microbial carbon (C), nitrogen (N) and phosphorus (P) pools were quantified in the organic horizon of soils from an arctic/alpine low-altitude heath and a high-altitude fellfield by the fumigation-extraction method before and after factorial addition of sugar, NPK fertilizer and benomyl, a fungicide. In unamended soil, microbial C, N and P made up 3.3–3.6%, 6.1–7.3% and 34.7% of the total soil C, N and P content, respectively. The inorganic extractable N pool was below 0.1% and the inorganic extractable P content slightly less than 1% of the total soil pool sizes. Benomyl addition in spring and summer did not affect microbial C or nutrient content analysed in the autumn. Sugar amendments increased microbial C by 15 and 37% in the two soils, respectively, but did not affect the microbial nutrient content, whereas inorganic N and P either declined significantly or tended to decline. The increased microbial C indicates that the microbial biomass also increased but without a proportional enhancement of N and P uptake. NPK addition did not affect the amount of microbial C but almost doubled the microbial N pool and more than doubled the P pool. A separate study has shown that CO₂ evolution increased by more than 50% after sugar amendment and by about 30% after NPK and NK additions to one of the soils. Hence, the microbial biomass did not increase in response to NPK addition, but the microbes immobilized large amounts of the added nutrients and, judging by the increased CO₂ evolution, their activity increased. We conclude: (1) that microbial biomass production in these soils is stimulated by labile carbon and that the microbial activity is stimulated by both labile C and by nutrients (N); (2) that the microbial biomass is a strong sink for nutrients and that the microbial community probably can withdraw substantial amounts of nutrients from the inorganic, plant-available pool, at least periodically; (3) that temporary declines in microbial populations are likely to release a flush of inorganic nutrients to the soil, particularly P of which the microbial biomass contained more than one third of the total soil pool; and (4) that the mobilization-immobilization cycles of nutrients coupled to the population dynamics of soil organisms can be a significant regulating factor for the nutrient supply to the primary producers, which are usually strongly nutrient-limited in arctic ecosystems.     

Nitrogen-fixing bacteria of the genusBeijerinckia in South African soils

Summary In contrast to the ubiquitous distribution of the nitrogen-fixing bacteria of the genusAzotobacter which can be encountered in all soils with suitable pH, the nitrogen-fixing bacteria of the genusBeijerinckia seem to be restricted to the tropics.All attempts to isolateBeijerinckia from north-european soils have so far failed. The conditions underlying this peculiar phenomenon are not clearly understood. The selective action of low soil pH, low temperature, frost resistance, etc. cannot be responsible for the absence ofBeijerinckia from temperate zones.In an earlier paper¹³ it was emphasized that the presence ofBeijerinckia is almost invariably connected with the occurrence of laterites, tropical Red Earths or latosols. As these soil types are almost restricted to the tropics, this may be the reason why the genusBeijerinckia is mainly confined to tropical regions.It was also suggested in that paper thatBeijerinckia might also occur in lateritic soils — fossil or developed under particular climatic conditions — outside the tropics.The present paper gives the first evidence of the occurrence ofBeijerinckia in lateritic soils outside the tropics in South Africa. ThereBeijerinckia occurs abundantly in the lateritic soils of the Mistbelt, but it occurs also — although more sporadically — in soils as far south as Cape Town (lat. 34° S.). Hence, non-symbiotic nitrogen fixation may occur in some of these soils which are too acid to containAzotobacter.     

Contrasting responses to ectomycorrhizal inoculation in seedlings of six tropical African tree species

Five caesalpinioid legumes, Afzelia africana, Afzelia bella, Anthonotha macrophylla, Cryptosepalum tetraphylum and Paramacrolobium coeruleum, and one Euphorbiaceae species, Uapaca somon, with a considerable range in seed sizes, exhibited different responses to inoculation by four species of ectomycorrhizal (ECM) fungi, Scleroderma dictyosporum, S. verrucosum, Pisolithus sp. and one thelephoroid sp. in greenhouse conditions. Thelephoroid sp. efficiently colonized seedlings of all of the five caesalpinioid legumes except U. somon, but provided no more growth benefit than the other fungi. Thelephoroid sp. and S. dictyosporum colonized seedlings of U. somon poorly, but stimulated plant growth more than the other fungi. The relative mycorrhizal dependency (RMD) values of the caesalpinioid legumes were never higher than 50%, whilst U. somon had RMD values ranging from 84.6 to 88.6%, irrespective of the fungal species. The RMD values were negatively related to seed mass for all plant species. Potassium concentrations in leaves were more closely related than phosphorus to the stimulation of seedling biomass production by the ECM fungi. Our data support the hypothesis that African caesalpinioid legumes and euphorbe tree species with smaller seeds show higher RMD values than those with the larger seeds.     

Biogeochemical responses of two forest streams to a 2-month calcium addition

1. Calcium (Ca) has been lost from forest soils at the Hubbard Brook Experimental Forest (HBEF) because of decreased atmospheric input of Ca and high input of acid anions. Through time, this Ca loss has led to low streamwater Ca concentration and this change may affect stream ecosystem processes.
2. To test both the biogeochemical response of streams to increased calcium concentration and the role of streams in retaining calcium lost from soils, we added c. 120 μeq Ca L^?1 as CaCl₂ to two second‐order streams at HBEF for 2 months. One stream (buffered) also received an equivalent amount of NaHCO₃ to simulate the increase in pH and alkalinity if Ca were added with associated HCO₃^? ion. The other stream (unbuffered) received only CaCl₂. We collected water samples along a transect above and below the addition site at 11 dates: two before, seven during, and two after the addition.
3. Increase in pH in the buffered stream ranged from 5.6 to about 7.0 in the treated section. There was a net uptake of Ca on all sampling dates during the addition and these uptake rates were positively related to pH. Between 10 and 50% of the added Ca was taken up during the release in the 80‐m study reach. In the unbuffered stream, there was net uptake of Ca on only two dates, suggesting lower Ca uptake.
4. Water samples collected after the addition was stopped showed that a small fraction of the added Ca desorbed from sediments; the remainder was apparently in longer‐term storage in the sediments. No Ca desorbed from the stream sediments in the unbuffered stream, showing that sorption/desorption may be controlled by a pH‐induced increase in the number of exchange sites.
5. These streams appeared to be a significant sink for Ca over a 2‐month time scale, and thus, change in streamwater Ca during a year may be due to processing of Ca within the stream channel, as well as to changes in inputs from the catchment.     

Microbial content of commercial South African high-moisture dried fruits

AIM: The aim was to evaluate commercially available South African high-moisture dried fruits (HMDF) for the microbial, moisture and SO2 contents, as well as aw and pH. METHODS AND RESULTS: The microbial content of commercially available HMDF was evaluated using nine different growth media. The moisture content, aw) SO2 and pH of each product were determined using standard analytical methods. It was found that the highest total aerobic counts were generated from high-moisture dried (HMD) prunes and raisins. The most frequent spoilers were members of the genus Bacillus. Fungal counts were also very high in the apricot products, exceeding the limit of 1000 CFU g(-1) as set by HMDF producers. Members of the genus Staphylococcus were found in the HMD raisins and Salmonella and thermoduric organisms were isolated from the HMD prunes. CONCLUSIONS: The microbial levels of South African HMDF were within the limits set, with the exception of apricots. SIGNIFICANCE AND IMPACT OF STUDY: The study shows the presence of Salmonella, Staphylococcus and Clostridium in South African HMDF. The presence of thermoduric organisms indicated that the current pasteurization process is not adequate and that the addition of preservatives would be an additional method to ensure safety and quality.     

Microbial activity and soil respiration under nitrogen addition in Alaskan boreal forest

Climate warming could increase rates of soil organic matter turnover and nutrient mineralization, particularly in northern high‐latitude ecosystems. However, the effects of increasing nutrient availability on microbial processes in these ecosystems are poorly understood. To determine how soil microbes respond to nutrient enrichment, we measured microbial biomass, extracellular enzyme activities, soil respiration, and the community composition of active fungi in nitrogen (N) fertilized soils of a boreal forest in central Alaska. We predicted that N addition would suppress fungal activity relative to bacteria, but stimulate carbon (C)‐degrading enzyme activities and soil respiration. Instead, we found no evidence for a suppression of fungal activity, although fungal sporocarp production declined significantly, and the relative abundance of two fungal taxa changed dramatically with N fertilization. Microbial biomass as measured by chloroform fumigation did not respond to fertilization, nor did the ratio of fungi : bacteria as measured by quantitative polymerase chain reaction. However, microbial biomass C : N ratios narrowed significantly from 16.0 ± 1.4 to 5.2 ± 0.3 with fertilization. N fertilization significantly increased the activity of a cellulose‐degrading enzyme and suppressed the activities of protein‐ and chitin‐degrading enzymes but had no effect on soil respiration rates or ¹⁴C signatures. These results indicate that N fertilization alters microbial community composition and allocation to extracellular enzyme production without affecting soil respiration. Thus, our results do not provide evidence for strong microbial feedbacks to the boreal C cycle under climate warming or N addition. However, organic N cycling may decline due to a reduction in the activity of enzymes that target nitrogenous compounds.     

Microbial diversity of topographical gradient profiles in Fushan forest soils of Taiwan

To evaluate the microbial diversity of Fushan forest soils, the variation of soil properties, microbial populations, and soil DNA with soil depth in three sites of different altitude were analyzed. Microbial population, moisture content, total organic carbon (C_org), and total nitrogen (N_tot) decreased with increasing soil depth. The valley site had the lowest microbial populations among the three tested sites due to the low organic matter content. Bacterial population was the highest among the microbial populations. The ratios of cellulolytic microbes to the total bacteria in organic layers were high, implying their roles in the carbon cycle. The microbial biomass carbon (C_mic) and nitrogen (N_mic) contents ranged from 130.5 to 564.1 μg g⁻¹ and from 16.7 to 95.4 μg g⁻¹, respectively. The valley had the lowest C_mic and N_mic. The organic layer had the highest C_mic and N_mic and decreased with soil depth. Analysis using denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction (PCR) amplicons of 16S rDNA showed that the bacterial diversity of the three sites were very similar to each other in the major bands, and the variation was in the minor bands. However, the patterns in PCR-DGGE profile through gradient horizons were different, indicating the prevalence of specific microbes at different horizons. These results suggest that the microbial diversity in the deeper horizons is not simply the diluted analogs of the surface soils and that some microbes dominate only in the deeper horizons. Topography influenced the quantity and diversity of microbial populations.     

Resprouters vs reseeders in South African forest trees; a model based on forest canopy height

1. We expect tree species that regenerate primarily by sprouting to produce fewer seedlings than co-occurring species that regenerate mainly from seedlings, because of the trade-off between allocating resources either to ensuring vegetative reproduction (e.g. protective bark/latent buds) or to sexual reproduction (e.g. seeds).
2. Furthermore, resprouting species, because of their multi-stemmed nature, should be at a relative disadvantage, and therefore relatively infrequent, in tall forests. This is because a resprouting individual allocates resources to a number of basal branches/stems and buds rather than maximizing vertical extension of a single leader, as is the case in a seeder. Also, many tall stems arising from the same multi-stemmed base, as is the case in resprouters, will be relatively poorly supported in comparison to the single stem of a reseeder.
3. To test these two ideas we surveyed a number of plots in a range of South African forests and thicket communities. We noted the numbers of seedlings and resprouts for each species and determined a mean for each site.
4. Short forests and thickets were dominated by multi-stemmed species and there was an almost total absence of seedlings. In contrast, tall forests were dominated by single-stemmed reseeding species and were accompanied by seedlings.     

Microbial responses to environmental arsenic

Microorganisms have evolved dynamic mechanisms for facing the toxicity of arsenic in the environment. In this sense, arsenic speciation and mobility is also affected by the microbial metabolism that participates in the biogeochemical cycle of the element. The ars operon constitutes the most ubiquitous and important scheme of arsenic tolerance in bacteria. This system mediates the extrusion of arsenite out of the cells. There are also other microbial activities that alter the chemical characteristics of arsenic: some strains are able to oxidize arsenite or reduce arsenate as part of their respiratory processes. These type of microorganisms require membrane associated proteins that transfer electrons from or to arsenic (AoxAB and ArrAB, respectively). Other enzymatic transformations, such as methylation-demethylation reactions, exchange inorganic arsenic into organic forms contributing to its complex environmental turnover. This short review highlights recent studies in ecology, biochemistry and molecular biology of these processes in bacteria, and also provides some examples of genetic engineering for enhanced arsenic accumulation based on phytochelatins or metallothionein-like proteins.     

Unexpected microbial metabolic responses to elevated temperatures and nitrogen addition in subarctic soils under different land uses

Biogeochemistry - Subarctic regions are particularly affected by global warming. As vegetation periods lengthen, boreal forests could gradually be converted into agricultural land. How land use...     

The impact of long-term nitrogen addition on microbial community composition in three Hawaiian forest soils

We evaluated the microbial communities in three Hawaiian forest soils along a natural fertility gradient and compared their distinct responses to long-term nitrogen (N) additions. The sites studied have the same elevation, climate, and dominant vegetation, but vary in age of development, and thus in soil nutrient availability and nutrient limitation to plant growth. Fertilized plots at each site have received 100 kg ha year(-1) N addition for at least 8 years. Soil parameters, water content, pH, and ammonium and nitrate availability differed by site, but not between control and N-addition treatments within a site at the time of sampling. Microbial biomass also varied by site, but was not affected by N addition. In contrast, microbial community composition (measured by phospholipid analysis) varied among sites and between control and N-addition plots within a site. These data suggest that microbial community composition responds to N addition even when plant net primary productivity is limited by nutrients other than N. This may have implications for the behavior of forests impacted by atmospheric N deposition that are considered to be "nitrogen saturated," yet still retain N in the soil.     

Herbivorous insect species in the tree canopy of a relict South African forest

Abstract.

• 1 The herbivorous insects on twelve species of evergreen broadleafed trees were repeatedly sampled over a period of 11 months in a small relict forest on the east coast of South Africa. This extraordinarily speciose forest patch has an unusually high proportion of endemic tree species, some of which are extremely rare.
• 2 The insect herbivore fauna (number of species) seems to be markedly depauperate compared to that reported on native, broadleafed trees from other parts of the world. Some possible reasons for this are discussed.
• 3 The total number of herbivorous insect species on each tree species was strongly correlated with the local relative abundance of the host plant species.
• 4 There was no relationship between the total number of insect herbivore species on each tree species and the relative taxonomic isolation of the trees. The proportion of seemingly unique (= specialist) herbivorous insect species (i.e. those that occurred on one tree species only) was greatest on taxonomically isolated trees.
• 5 A fundamental deficiency in the interpretation of the data in this study, and of many other similar studies that report on the number of insect species on plants, is discussed, namely the lack of clarity on the closeness of the association between individual insect herbivore species and their respective host plants.

     

Microbial responses to salt-induced osmotic stress

Summary Soil columns were exposed to balanced (low Na⁺) or unbalanced (high Na⁺) high-salt solutions for a period of 7 days followed by 7 days of stress reflief. Total numbers of bacteria released into the perfusates rose under both types of stress, but the proportion of displaced bacteria that were viable fell significantly. Relief from both types of stress stimulated rapid increases in the number of viable micro-organisms released from soil. Examination of the soils at the end of the relief periods revealed that soils exposed to stress contained more viable bacteria than the non-stressed controls. However, high levels of balanced stress led to a significant decrease in species diversity within the microbial population, but a similar effect was not observed in soils exposed to unbalanced, high Na⁺ stress. These results suggest that, while salt stress may cause a significant reduction in the number of microorganisms in a soil, a large portion of the microbial population can rapidly adapt to marked changes in salinity.     

Nitrification in coniferous forest soils

Net nitrification rates tend to be low or negligible in the forest floor of many coniferous forests of North-East Scotland. The most likely process controls are substrate availability, pH, allelopathy, water potential, nutrient status and temperature. These are discussed in relation to field and laboratory studies of net and potential rates of nitrification.Fungi make up by far the largest part of the nitrifier community in the coniferous forest floor. Very little is known about the distribution and activity of autotrophs in these systems, although it is certain that in vitro evidence suggesting autotrophs cannot nitrify at pH levels characteristic of coniferous forest soils is unrealistic.Because of the metabolic diversity of nitrifying fungi, a variety of organic and inorganic nitrification pathways may exist in coniferous forests. The possible involvement of free radicles in fungal nitrification in coniferous forest soils is also suggested.A complete understanding of nitrification in coniferous forest soils can only result from field characterisation of N flux such as through the use of ¹⁵N. This must be combined with ecophysiological characterisation of the organisms involved in order that the complexity of nitrification in coniferous forest soils can be resolved.     

Threshold responses of soil organic carbon concentration and composition to multi-level nitrogen addition in a temperate needle-broadleaved forest

Responses of soil organic carbon (SOC) cycling and C budget in forest ecosystems to elevated nitrogen (N) deposition are divergent. Little is known about the N critical loads for the shift between gain and loss of SOC storage in the old-growth temperate forest of Northeast China. The objective of this study was to investigate the nonlinear responses of SOC concentration and composition to multiple rates of N addition, as well as the microbial mechanisms responsible for SOC alteration under N enrichment. Nine rates of urea addition (0, 10, 20, 40, 60, 80, 100, 120, 140 kg N ha^?1 year^?1) with 4 replicates for each treatment were conducted. Soil samples in the 0–10 cm mineral layer were taken after 3 years of N fertilization. Soil aggregate size distribution and SOC physical fractionation were performed to examine SOC dynamics. Phospholipid fatty acid (PLFA) technique was used to measure the abundance and structure of microbial community. Three years of N addition led to significant increases in the concentrations of soil particulate organic C and aggregate-associated organic C fractions only. The responses of total N and each labile SOC fraction to the rates of N addition followed Gaussian equations, with the N critical loads being estimated to be between 80 and 100 kg N ha^?1 year^?1. The change in SOC concentration (ΔSOC) was positively correlated with the changes in aggregate associated OC (r² > 0.80) and POC concentrations (r² > 0.50). Significant correlations among the concentrations of labile SOC fractions, the percentages of soil aggregates, and the abundances of microbial PLFAs were observed, which implies a close linkage between microbial community structure and SOC accumulation and stability. Our results suggest that increase in soil moisture and shift of microbial community structure could control the critical N load for the switch between C accumulation and loss. The current N deposition rate (~ 11 kg N ha^?1 year^?1) to the northeast China’s forests is favorable for soil C accumulation over the short term.