Comparison of silica gel plating and most probable number methods for counting nitrifiers in soils

Abstract

A silica gel plating method for counting of nitrifying microorganisms in soils was found equally satisfactory to most probable number method.     

Growth rates of Aporrectodea caliginosa (Oligochaetae: Lumbricidae) as influenced by soil temperature and moisture in disturbed and undisturbed soil columns

Earthworm growth is affected by fluctuations in soil temperature and moisture and hence, may be used as an indicator of earthworm activity under field conditions. There is no standard methodology for measuring earthworm growth and results obtained in the laboratory with a variety of food sources, soil quantities and container shapes cannot easily be compared or used to estimate earthworm growth in the field. The objective of this experiment was to determine growth rates of the endogeic earthworm Aporrectodea caliginosa (Savigny) over a range of temperatures (5–20 °C) and soil water potentials (−5 to−54 kPa) in disturbed and undisturbed soil columns in the laboratory. We used PVC cores (6 cm diameter, 15 cm height) containing undisturbed and disturbed soil, and 1 l cylindrical pots (11 cm diameter, 14 cm height) with disturbed soil. All containers contained about 500 g of moist soil. The growth rates of juvenile A. caliginosa were determined after 14–28 days. The instantaneous growth rate (IGR) was affected significantly by soil moisture, temperature, and the temperature×moisture interaction, ranging from −0.092 to 0.037 d⁻¹. Optimum growth conditions for A. caliginosa were at 20 °C and −5 kPa water potential, and they lost weight when the soil water potential was −54 kPa for all temperatures and also when the temperature was 5 °C for all water potentials. Growth rates were significantly greater in pots than in cores, but the growth rates of earthworms in cores with undisturbed or disturbed soil did not differ significantly. The feeding and burrowing habits of earthworms should be considered when choosing the container for growth experiments in order to improve our ability to extrapolate earthworm growth rates from the laboratory to the field.     

Comparison of NH4 pool dilution techniques to measure gross N fluxes in a coarse textured soil

We compared gross N fluxes by ¹⁵N pool dilution in a coarse-textured agricultural soil when ¹⁵N was applied to the soil NH₄⁺ pool by either: (i) mixing a ¹⁵NH₄NO₃ solution into disturbed soil or (ii) injection of ¹⁵NH₃ gas into intact soil cores. The two techniques produced similar results for gross N mineralization rates indicating that NH₄⁺ production in soil was not altered by soil disturbance, method of application (gas vs. solution), or amount of N applied. This was not the case for immobilization rates, which were twofold higher when ¹⁵N label was applied to the soil NH₄⁺ pool with the mixing technique compared to the injection technique. This was attributed to the fact that more NH₄⁺ was applied with the mixing technique. Estimates of gross nitrification were accompanied by large error terms meaning differences between ¹⁵N labeling methods could not be accurately assessed for this process rate.     

Growth of Lumbricus terrestris and Aporrectodea caliginosa in an acid forest soil, and their effects on enchytraeid populations and soil properties

A laboratory experiment was carried out to test the hypothesis that the earthworms Lumbricus terrestris and Aporrectodea caliginosa are able to maintain their populations and reproduce in the acid forest soil of a deciduous forest where no lumbricids were found in the field. The experiment was conducted in 45-l containers in which layers of mineral subsoil, humus and organic topsoil collected from the site were established. Both species survived and at least L. terrestris reproduced during the 60 weeks’ incubation. Burrows and middens of L. terrestris were recorded and quantities of litter were consumed. The presence of lumbricids increased the organic matter content of humus, reduced the acidity of the topsoil and humus layers, and suppressed the population of the enchytraeid Cognettia sphagnetorum. A dense population of Enchytraeus albidus was found in L. terrestris middens. It is concluded that edaphic factors do not explain the absence of earthworms, but isolation from cultural landscapes and lack of opportunity to colonize the site from the surroundings is the decisive factor.     

Responses of co-occurring populations of Dendrobaena octaedra (Lumbricidae) and Cognettia sphagnetorum (Enchytraeidae) to soil pH, moisture and resource addition

Dendrobaena octaedra (Lumbricidae) and Cognettia sphagnetorum (Enchytraeidae) are the two most dominating soil invertebrates in terms of biomass in boreal coniferous forest soils. A microcosm experiment was set up in order to study the influence of pH, moisture and resource addition on D. octaedra and C. sphagnetorum when both species are simultaneously present. Two kinds of coniferous forest humus were used as substrate, pine stand humus (pH 4.2), and spruce stand humus (pH 4.6); in the third treatment the pine stand humus was adjusted with slaked lime (CaOH₂) to the same initial pH as the spruce stand humus. Each substrate was adjusted to water contents of 25%, 42.5% and 60% of WHC (referred to as ‘dry’, ‘moist’ and ‘wet’). In the second part of the experiment, spruce needle litter and birch leaf litter were separately added into the pine stand humus (‘moist’, unlimed) and compared with a control without litter. The microcosms were plastic jars with 75 g (d.m.) of humus, into which 4 specimens of D. octaedra and 70 specimens of C. sphagnetorum were added. D. octaedra showed the highest biomass and C. sphagnetorum the lowest biomass in the spruce stand humus with higher pH. Moisture did not affect earthworms, while C. sphagnetorum thrived best at the highest moisture. Addition of both kinds of litter increased the numbers and biomass of D. octaedra, while on C. sphagnetorum resource addition had little effect. The results help to explain the abundance of these two species in coniferous forests differing in soil acidity, moisture and fertility.     

14C-labelled maleic hydrazide degradation in soil: Influence of temperature,moisture, clay and organic material

¹⁴C-labelled maleic hydrazide (MH) was added to each of three soils at a concentration of 4 mg kg^?1, and its degradation measured by the release of ¹⁴CO₂ after 2 days. Between 1 and 30°C, at a constant moisture content (full field capacity), the mean degradation rate increased by a factor of 3 for each temperature increment of 10°C (Q₁₀ = 3). The mean activation energy was 78 kJ mol^?1. Above 35°C, the degradation rate decreased.At soil moisture contents between wilting point and 80–90% of field capacity, the degradation rate doubled with an increase in moisture content of 50% of field capacity (constant temperature, 25°C). Above field capacity, the degradation rate was either unchanged or decreased. Below wilting point the degradation was very slow, even after 2 months.The rate of decomposition of MH at all temperatures and moisture contents was lowest in the soil with the highest content of organic matter and the lowest clay content. This soil had the highest Freundlich K value, and presumably adsorbed MH the most strongly, although the lower clay content may also play a role in the lower decomposing capacity of this soil.     

Interactive effect of tillage depth and mulch on soil temperature, productivity and water use pattern of rainfed barley (Hordium vulgare L.)

Soil porosity and organic matter content influence the hydrology, thermal status and productivity of agricultural soils. Shape, size and continuity of soil pores are determined by tillage practices. Thus appropriate tillage and mulch management can conserve residual soil moisture during the post rainy season. This can play a key role in enhancing productivity under the rainfed ecosystem of subhumid region in eastern India. A field study was carried out on a fine loamy soil from 1993–1994 to 1995–1996. Two tillage treatments were conventional ploughing (150 mm depth) and shallow ploughing (90 mm) depth. Each tillage practice was tested with three mulch management viz., no mulch, soil dust mulch and rice (Oryza sativa L.) straw mulch. Soil organic carbon, bulk density, moisture retentivity, soil temperature with productivity and water use pattern of barley (Hordium vulgare L.) were measured.Reduction in ploughing depth resulted in nominal increase in profile (0.0–1.2 m) moisture status, yield, and soil thermal status at 14:00 and water use efficiency (WUE). However, it decreased the magnitude of soil temperature in the morning (07:00). Straw mulch conserved 19–21 mm of moisture in the profile (1.2 m) over the unmulched condition. Both soil dust and rice straw mulching elevated soil thermal status at 07:00 as compared to unmulched condition, but this trend was reversed at 14:00. Straw mulching significantly increased grain yield and WUE over soil dust mulch and unmulched condition. Impact of straw mulch was more pronounced under shallow ploughing depth. Shallow tillage with rice straw mulching is recommended to the farmers to obtain higher level of yield and water use efficiency.     

Comparison of net ecosystem CO2 exchange in two peatlands in western Canada with contrasting dominant vegetation, Sphagnum and Carex

Net ecosystem carbon dioxide exchange was measured in two contrasting peatlands in northern Alberta, Canada using the eddy covariance technique during the growing season (May–October). Sphagnum spp. made up approximately 66% of the total LAI (1.52 m² m⁻²) at the poor fen and the total N content of Sphagnum capitula was 7.8 mg g⁻¹ at the peak of the growing season. In contrast, the dominant plant species at the extreme-rich fen site, the perennial sedge, Carex lasiocarpa, accounted for approximately 60% of the total LAI (1.09 m² m⁻²), and had leaf total N content of 19.3 mg g⁻¹ at peak biomass. In addition, the peak aboveground biomass was higher at the poor fen (230.9 g m⁻²) than at the extreme-rich fen (157.1 g m⁻²). Both sites had maximum daily rates of net CO₂ uptake of approximately 5 μmol m⁻² s⁻¹, and typical nighttime rates of CO₂ loss of approximately 2 μmol m⁻² s⁻¹ during the peak of the growing season. Calculations of maximum photosynthetic and respiratory capacity were consistently higher at the extreme-rich fen. The poor fen was a net sink for CO₂ during 4 of the 6 months (peaking at 44 g C m⁻² in July), while only slight net losses of CO₂ (3 g C m⁻²) occurred in May and September. In contrast, the extreme-rich fen was calculated to be a significant net sink for CO₂ only during 2 months of the growing season (peaking at 30 g C m⁻² in August), while significant net losses of CO₂ occurred in May (8 g C m⁻²) and in October (13 g C m⁻²). The plant species at the poor fen site were active earlier and later in the growing season, while it took longer for C. lasiocarpa to develop leaf tissue, and leaf senescence and reduction in photosynthetic activity occurred earlier in the fall at the extreme-rich fen. When integrated over the 6-month growing season, the poor fen was a net sink (90 g C m⁻²) that was three times larger than the extreme-rich fen (31 g C m⁻²). The ratio of cumulative total ecosystem respiration to gross primary production was 0.7 at the poor fen and 0.9 at the extreme-rich fen.     

Growth of Gaeumannomyces graminis var. tritici in soil: Effects of temperature and water potential

The effects of temperature and water on the growth of the take-all fungus, Gaeumannomyces graminis var. tritici (Ggt), were examined in two factorial experiments. The first examined the effects of temperature and water potential on the growth of two isolates of Ggt on agar media, using osmotically-adjusted water potentials. The second experiment was concerned with the growth of the Ggt isolates in one sterile and two natural soils at two water regimes in the absence of a living host. Three temperatures (10, 18 and 26°C) were used in these experiments. A third experiment determined growth through soil.Growth was greatest at high temperatures and low water potential in axenic culture, but in unsterile soil growth at different temperatures and water potentials was strongly influenced by competition from the soil biota. The best temperature for growth in unsterile soil was 18°C. Growth at 26°C in unsterile soil was greatly reduced, this being attributed to more intense microbial competition. In sterile soil Ggt grew equally well at 18 and 26°C. At 10°C, both isolates of Ggt grew better in unsterile soil than in sterile soil.Under suitable conditions Ggt grew out readily from infected straw into unsterile soil (up to 5 cm in 10 days) in the absence of a host plant, forming melanized, hyaline and branched hyphae. These hyphae were infectious after dry storage for 5 months in the laboratory. Ggt thus appears to be a more successful soil inhabitant than is widely believed. Our experiments could explain many of the host-based concepts related to field expression of disease.The technique presented here could be of value for testing the suppressiveness or conduciveness of soils by measuring fungal growth in soil.     

Relationships among fine roots, fungal hyphae and soil microarthropods among different soil microhabitats in a temperate coniferous forest of Chamaecyparis obtusa

The relationships between roots and soil communities are not well understood. We used the ingrowth-core method with L-, FH-, and M-layer substrates to investigate the relationships among soil organic carbon, fine root biomass, hyphal length and the numbers of soil microarthropods. The study was carried out in a temperate forest of the arbuscular mycorrhizal conifer, Chamaecyparis obtusa. The relationships among fine roots, fungi and soil microarthropods were different among soil substrates and faunal taxa. Soil carbon contents, fine root biomass, hyphal length and soil-microarthropod numbers were the highest in the FH-substrate, and the lowest in the M-substrate. For each substrate, the total numbers of soil microarthropods did not positively correlated with soil organic carbon. A positive correlation between fine root biomass and the soil microarthropod numbers was significant only in the M-substrate, but not in the L- and FH-substrates. In M-substrates, strong positive correlations were found between fine root biomass or hyphal length and Mesostigmata or Oribatida numbers, but Collembola numbers were not corelated. Further studies of the regulation mechanism of soil food web structures should note that the soil microarthropods have different responses to C sources according to soil conditions and trophic interactions.     

Response of mycorrhizal, rhizosphere and soil basal respiration to temperature and photosynthesis in a barley field

The mycorrhizal, rhizosphere and basal components of soil respiration were partitioned in a barley field experiment with the main objective of determining the controlling effects of photosynthetic activity and temperature on soil respiration sources. Micro-pore meshes were used to create both root and mycorrhiza-free soil cores over which collars for soil respiration measurements were inserted. Differences between mesh treatments were used to determine the contribution of each component. With a focus on the growing season, we analyzed the response of respiration sources to photosynthesis, temperature and moisture, as well as changes in microbial biomass, mineral nitrogen and carbon-nitrogen ratios responding to treatment and time of year. Results gave clear differences between sources in their response to both temperature and photosynthetic activity and showed that several processes are involved in determining respiration rates as well as apparent temperature relations. In particular, the respiration of arbuscular mycorrhizal hyphae was seen to be a significant amount of root derived carbon respiration (25.3%) and consequently of total assimilated carbon (4.8%). This source showed a stronger response to photosynthetic activity than the rhizosphere component (r²=0.79, p<0.001 and r²=0.324, p=0.53, respectively). Q₁₀ values—the increase in respiration rates with a 10 °C increase in temperature—changed seasonally and showed temperature relations being dependent on the presence of mycorrhizal and rhizosphere respiration sources, as well as on plant development. Respiration from mycorrhizal hyphae and the rhizosphere showed no response (r²=0, p<0.99) or low response (r²=0.14, p<0.01) to temperature, respectively. We conclude that the potential importance and controls of mycorrhizal fungi respiration in croplands are comparable to those observed in other ecosystems, and that temperature response curves should be carefully interpreted given that substrate availability and plant dynamics strongly regulate respiration rates in ecosystems.     

Short-term carbon and nitrogen dynamics in soil,litterfall and canopy of a suburban native forest subjected to prescribed burning in subtropical Australia

Purpose

This study aimed to understand the mechanisms of the variations in carbon (C) and nitrogen (N) pools and examine the possibility of differentiating the burning effects from seasonal and pre-existed N limitations in a native suburban forest ecosystem influenced by prescribed burning in subtropical Australia.

Materials and methods

Soil and litterfall samples were collected from two study sites from 1 to 23 months since last burnt. Soil labile C and N pools, soil C and N isotopic compositions (δ¹³C and δ¹⁵N), litterfall mass production (LM), and litterfall total C, total N, δ¹³C and δ¹⁵N were analysed. In-situ gas exchange measurements were also conducted during dry and wet seasons for Eucalyptus baileyana and E. planchoniana.

Results and discussion

The results indicated that labile C and N pools increased within the first few months after burning, with no correlations with climatic factors. Therefore, it was possible that the increase was due to the burning-induced factors such as the incorporation of ashes into the soil. The highest values of soil and litterfall δ¹⁵N, observed when the study was commenced at the experimental sites, and their high correlations with climatic factors were indicative of long-term N and water limitation. The ¹³C signals showed that soil N concentrations and climatic factors were also two of the main factors controlling litterfall and foliage properties mainly through the changes in photosynthetic capacity and stomatal conductance.

Conclusions

Long-term soil N availabilities and climatic factors were the two of the main driving factors of C and N cycling in the studied forest sites. Further studies are needed to compare soil and litterfall properties before and after burning to profoundly understand the effects of prescribed burning on soil labile C and N variations.

     

Labile, recalcitrant, and microbial carbon and nitrogen pools of a tallgrass prairie soil in the US Great Plains subjected to experimental warming and clipping

Carbon (C) and nitrogen (N) fluxes are largely controlled by the small but highly bio-reactive, labile pools of these elements in terrestrial soils, while long-term C and N storage is determined by the long-lived recalcitrant fractions. Changes in the size of these pools and redistribution among them in response to global warming may considerably affect the long-term terrestrial C and N storage. However, such changes have not been carefully examined in field warming experiments. This study used sulfuric acid hydrolysis to quantify changes in labile and recalcitrant C and N fractions of soil in a tallgrass prairie ecosystem that had been continuously warmed with or without clipping for about 2.5 years. Warming significantly increased labile C and N fractions in the unclipped plots, resulting in increments of 373 mg C kg⁻¹ dry soil and 15 mg N kg⁻¹ dry soil, over this period whilst clipping significantly decreased such concentrations in the warmed plots. Warming also significantly increased soil microbial biomass C and N in the unclipped plots, and increased ratios of soil microbial/labile C and N, indicating an increase in microbial C- and N-use efficiency. Recalcitrant and total C and N contents were not significantly affected by warming. For all measured pools, only labile and microbial biomass C fractions showed significant interactions between warming and clipping, indicating the dependence of the warming effects on clipping. Our results suggest that increased soil labile and microbial C and N fractions likely resulted indirectly from warming increases in plant biomass input, which may be larger than warming-enhanced decomposition of labile organic compounds.     

A comparison of two colorimetric assays,based upon Lowry and Bradford techniques,to estimate total protein in soil extracts

Soil extracts usually contain large quantities of dissolved humified organic material, typically reflected by high polyphenolic content. Since polyphenols seriously confound quantification of extracted protein, minimising this interference is important to ensure measurements are representative. Although the Bradford colorimetric assay is used routinely in soil science for rapid quantification protein in soil-extracts, it has several limitations. We therefore investigated an alternative colorimetric technique based on the Lowry assay (frequently used to measure protein and humic substances as distinct pools in microbial biofilms). The accuracies of both the Bradford assay and a modified Lowry microplate method were compared in factorial combination. Protein was quantified in soil-extracts (extracted with citrate), including standard additions of model protein (BSA) and polyphenol (Sigma H1675-2). Using the Lowry microplate assay described, no interfering effects of citrate were detected even with concentrations up to 5 times greater than are typically used to extract soil protein. Moreover, the Bradford assay was found to be highly susceptible to two simultaneous and confounding artefacts: 1) the colour development due to added protein was greatly inhibited by polyphenol concentration, and 2) substantial colour development was caused directly by the polyphenol addition. In contrast, the Lowry method enabled distinction between colour development from protein and non-protein origin, providing a more accurate quantitative analysis. These results suggest that the modified-Lowry method is a more suitable measure of extract protein (defined by standard equivalents) because it is less confounded by the high polyphenolic content which is so typical of soil extracts.     

Differential growth response of Curculigo orchioides to native arbuscular mycorrhizal fungal (AMF) communities varying in number and fungal components

The study reports diversity of arbuscular mycorrhizal fungal (AMF) species in the rhizosphere of an endangered anticancerous herb – Curculigo orchioides Gaertn. in its natural habitat. A total of 18 species of AMF, belonging to three genera (Acaulospora, Glomus and Gigaspora) were recorded, with Glomus microcarpum being the most abundant species type. The AMF species composition across the study sites appeared to be influenced by soil pH rather than soil P and vegetation. Acaulospora laevis spores were restricted to sites where the soil pH was acidic. The effectiveness of these native AMF species on growth performance of C. orchioides plants was compared under experimental conditions. In general, the mycorrhizal plants were superior in most of the evaluated parameters, but the extent to which the growth of mycorrhizal plants was influenced varied with the inocula used. The plants inoculated with mixed consortia containing maximum AMF species richness exhibited improved growth in comparison to consortia containing lower AMF diversity and monospecies cultures. The variable plant responses observed with any two consortia having same species richness in the present study could be due to variable component AMF species and their relative abundance. These results emphasize the need to protect the below-ground diversity of AMF and recommend their usage for restoration practices.     

Exploitation of litter and soil by the train millipede Parafontaria laminata (Diplopoda: Xystodesmidae) in larch plantation forests in Japan

Swarms of the train millipede (Parafontaria laminata) are known to occur every 8 years; during the swarms, adults emerging from the soil have an extremely high density in natural and plantation forests in central Japan. Their influence on organic layer accumulation on forest floors was investigated under field conditions in plantation larch forests in central Japan, before and after the adult swarming period of 2000. In addition, the adult millipede feeding preference and the amount of food consumed were observed in the laboratory. The field density of adults ranged from 11 to 311 individuals m⁻² in October 2000; the highest biomass was 28.6±16.4 g dry wt m⁻². Reduction of the forest floor organic layer was density- dependent when the population consisted of 7th instar larvae; however, the adult population did not consume in a density-dependent manner. Higher levels of geophagy were observed in high-density treatments in the laboratory. We hypothesized that the geophagy of the millipedes was partly supported by the naturally high organic matter content of Andosol, and that soil consumption would sustain an extremely high biomass in a temperate conifer forest soil.