Influence of storage on soil microbial biomass estimated by three biochemical procedures

The effects of 28 and 56 days' storage at 25°, 4° and ?20°C on the microbial biomass content of four soils from tussock grasslands were studied by three biochemical procedures. Two of the procedures involved measurement of CO₂ and mineral-N (Min-N) production by chloroform-fumigated and unfumigated soil, and consequent estimation of biomass C and Min-N flush respectively. In the third, adenosine 5'-triphosphate (ATP) content was determined.Patterns of CO₂ production were often influenced by storage treatment. The use of fixed incubation periods for estimating the CO₂ flush of fumigated soil and the steady rate of CO₂ production by unfumigated soil did, however, give biomass C estimates that were generally similar to those calculated from individually determined incubation periods for each treatment and soil.Biomass C values could change significantly at all storage temperatures, but generally least at ?20°C. Storage at ?20°C was also the most suitable for retaining ATP contents, whereas 4°C was best for values of Min-N flush. Values of Min-N flush after storage of soil at ?20°C decreased significantly in two of the soils but increased in another. No storage temperature was thus satisfactory for all three indices of microbial biomass. Generally, however, 4°C was adequate for short periods, and 25°C the least suitable.     

Microbial biomass estimations in soils from tussock grasslands by three biochemical procedures

Microbial biomass was determined by three biochemical procedures in nine topsoils from a climosequence in tussock grasslands. The pH values of the samples ranged from 4.4 to 6.2 and organic C contents from 2.5 to 20.0%. When determined by a chloroform-fumigation procedure, contents of biomass C and mineral-N (Min-N) flush ranged from 530–2780 and 59–167 μgg^?1 dry soil respectively. Adenosine 5'-triphosphate (ATP) content ranged from 2.2 to 10.7 μg g^?1 dry soil. All three estimates were significantly correlated with each other and with several soil properties, including organic C and total N contents and CO₂ production. They were not significantly correlated with any climatic factor.In spite of these significant correlations, the ratios of the biomass estimates varied appreciably in the different soils. The ratios of biomass C/Min-N flush ranged from 7.8 to 22.8 (average 12.5), biomass C/ATP from 163 to 423 (average 248) and Min-N flush/ATP from 12 to 35 (average 22). These ratios were mostly higher than those found elsewhere for Australian and English soils. The high biomass C/ATP and Min-N flush/ATP ratios did not appear to originate from inefficient extraction of “native” ATP or from the soils' P status. Based on these results, care in the use of factors for obtaining soil microbial biomass content from Min-N flush or ATP values is indicated.     

The effects of biocidal treatments on metabolism in soil—I. Fumigation with chloroform

This series of five papers is a study of how biocidal treatments influence metabolism in soil, directed particularly towards the flush of decomposition caused by fumigation, and designed to see if the size of this flush can be used as a measure of the soil biomass.Chloroform fumigation caused an immediate increase in the amounts of ammonium and organic C extracted from a soil by 1 N K₂SO₄. When the CHCl₃-treated soil was then inoculated with fresh soil and incubated for 10 days. it consumed 2·8 times more O₂, evolved 2·2 times more CO₂ and mineralised 7·3 times more N than an unfumigated soil. Extractable organic C decreased by about 40% when the fumigated soil was incubated for 10 days. A second fumigation given immediately after the first produced no further increase in the flush, but some recovery occurred if the soil was incubated between fumigations. However, this recovery was slow and incomplete; a second fumigation given 53 days after the first gave a flush only one-seventh the size of the first. Glucose (or ryegrass) added to the soil and allowed to decompose before fumigation increased the size of the flush. After a 52-day incubation, 29% of the C originally added as ¹⁴C labelled glucose remained in the soil; fumigation on the 52nd day increased the evolution of labelled CO₂ during the subsequent 10-day period by a factor of 8. Fumigation of a soil that had already been sterilized by 2·5 Mrads of gamma radiation increased the flush slightly; the amount of O₂ consumed in 10 days increased from 123 to 137 mg/100 g soil. It is proposed that the flush of decomposition following CHCl₃ fumigation is caused by the decomposition of killed organisms by the survivors (or by organisms added in the inoculum) and that organisms are more rapidly and completely attacked after exposure to CHCl₃ than after irradiation. On this hypothesis. 10% of the glucose C originally added to the soil was located in the soil biomass after 52 days.     

Mineralization of bacteria and fungi in chloroform-fumigated soils

It has been suggested by others that the size of the flush of mineralization caused by CHC1₃ fumigation can be used to estimate the amount of microbial biomass in soils. Calculation of biomass from the flush requires that the proportion of CHCl₃-killed cell C mineralized be known. To determine this proportion, 15 species of [¹⁴C]labelled fungi and 12 species of [¹⁴C]labelled bacteria were added to four types of soil and these were fumigated for 24 h with CHC1₃, reinoculated with unfumigated soil, and incubated at 22°C for 10 days. The average percentage mineralization of the fungi was 43.7 ± 5.3, while the average for the bacteria was 33.3 ± 9.9. Using a 1:3 ratio for distribution of total biomass between the bacterial and fungal populations, respectively, it was calculated that the average mineralization of both types of cells was 41.1%. In experiments conducted to determine if CHC1₃ vapour alters stabilized microbial metabolites or dead microbial cells in a manner which makes them more susceptible to degradation, it was found that both fumigated and unfumigated dead fungal materials mineralized to the same extent in soil during 10 days of incubation.     

Activity and degradation of streptomycin and cycloheximide in soil

Streptomycin and cycloheximide were added (3 and 2 mg g^-1 dry soil, respectively) single and in combination to a forest soil to follow their possible degradation and their effects on soil mineralization-immobilization processes. After 0, 1, 2, 4, 7, and 10 days of incubation at 25°C and 60% water-holding capacity, measurements were taken of microbial biomass C and N, the evolution of CO₂, exchangeable NH _inf4 ^sup+ , 0.5M K₂SO₄-extractable organic C, and total N in both unfumigated and CHCl₃-fumigated soil. The results indicated that during the first 2 days of incubation, soil microorganisms were killed by the antibiotics and/or by CHCl₃ and used subsequently as a substrate by the survivors. Thereafter, surviving microorganisms probably also started to use biocidal molecules as an energy and nutrient source. The ratios of biomass C to biomass N and of CO₂ evolved to net NH _inf4 ^sup+ produced indicated that both biocides had non-target effects for most of the incubation. Thus, streptomycin and cycloheximide are not suitable in determining the relative contribution from fungi and bacteria to mineralization-immobilization processes in soils.     

Microwave irradiation of soil for routine measurement of microbial biomass carbon

 Microwave irradiation was evaluated as a non-toxic alternate to chloroform fumigation for routine measurement of soil microbial biomass C. Microwave energy was applied to moist soil to disrupt microbial cells. The flush of C released was then measured after extraction or incubation. Microwave irradiation at 800 J g^–1 soil was optimal because this level resulted in an almost instantaneous rise in soil temperature (≥80  °C), an abrupt reduction in microbial activity, maximal release of biomass C, and minimal solubilization of humic substances. Both incubation-CO₂ titration and extraction-colorimetry methods were used on separate 20-g subsamples to compare the labile C in the microwave-treated and untreated soil samples. The incubation-titration method was also used to measure C in chloroform-fumigated soil samples. Averaged across soils, the chloroform fumigation yielded 123.3±5.1 mg CO₂-C kg^–1. Microwave irradiation yielded 93.6±3.9 mg CO₂-C kg^–1 soil determined by incubation and 52.4±2.4 mg C kg^–1 soil determined by extraction, accounting for 76% and 42% of the net flush of C measured by the chloroform fumigation. Microwave-stimulated net flushes of C were correlated closely (r ²=0.974 for incubation or 0.908 for extraction) with microbial biomass C measured by the chloroform fumigation. Little correlation was found with the total soil organic C (r ²=0.241 for incubation or for 0.166 extraction). Mean efficiency factors for incubation (K _MI) or extraction (K _ME) were used to calculate microbial biomass C from net flushes of C between microwaved and unmicrowaved soils. Values of K _MI and K _ME were not affected by soil pH, bulk density or clay contents. Extraction of microwaved soil by 0.5M K₂SO₄ proved to be a simple, fast, precise, reliable, and safe method to measure soil microbial biomass C. Received: 12 September 1997     

The effects of biocidal treatments on metabolism in soil—V: A method for measuring soil biomass

A new method for the determination of biomass in soil is described. Soil is fumigated with CHCl₃ vapour, the CHCl₃ removed and the soil then incubated. The biomass is calculated from the difference between the amounts of CO₂ evolved during incubation by fumigated and unfumigated soil. The method was tested on a set of nine soils from long-term field experiments. The amounts of biomass C ha^?1 in the top 23 cm of soil from plots on the Broadbalk continuous wheat experiment were 530 kg (unmanured plot), 590 (plot receiving inorganic fertilizers) and 1160 (plot receiving farmyard manure). Soils that had been fallowed for 1 year contained less biomass than soils carrying a crop. A calcareous woodland soil contained 1960 kg biomass C ha^?1, and an unmanured soil under permanent grass 2020. The arable soils contained about 2% of their organic C in the biomass; uncultivated soils a little more—about 3%.     

Application of eco-physiological quotients (qCO2 and qD) on microbial biomasses from soils of different cropping histories

Metabolic quotients for CO₂C (qCO₂C) and microbial-C-loss (qD) were studied on soil microbial communities under long-term monoculture (M) or continuous crop rotations (CR). Under defined laboratory conditions the mean qCO₂C (unit CO₂C unit⁻¹ C_mic h⁻¹) of different microbial biomasses from 17 M systems amounted to 1.097 μg CO₂qCO₂CC as compared to 0.645 μg CO₂C of microbial biomasses from 19 CR systems. The 1.7 times higher CO₂C release per unit biomass and time of microbial biomasses from M systems was significantly different at the P =0.001 level.In addition, microbial C-loss in samples from M or CR plots was followed for 5 weeks. Again, mean qD per unit microbial biomass and time was 1.6 times higher (P = 0.01) for microbial biomasses from M systems (0.301 μg C, 14 soils) when compared with CR systems (0.188μg C, 14 soils).These differences were not related to soil texture, C_org or pH of these soils. The effects of environmental influences (soil management) on the microbial pool in terms of a changing energy demand are discussed.     

Bacterial,fungal and protozoan responses to chloroform fumigation in stored soil

Microbial biomass estimated by CO₂ evolution following fumigation was 2.5–14.7 times greater than that estimated by direct microscopy in prairie soil. Bacteria, fungi and protozoa were counted by direct microscopy before, during and periodically for 10 days following chloroform fumigation and compared with microbial biomass as estimated by CO₂ evolution and N mineralization following chloroform fumigation. Protozoan populations were reduced to below detection levels immediately after fumigation and remained below detection levels during incubation following fumigation. Bacterial and fungal populations were reduced by fumigation to 37–79% of their original populations but usually recovered to their initial numbers by the second day following fumigation. In one case protozoa contributed up to 74 μg C, or about half of the total microbial biomass, to CO₂ evolution following fumigation.Microbial biomass was estimated in soil wetted to 60% of water-holding capacity (WHC) 1 wk or 1 day before fumigation. Microbial activity changed during the 1 wk incubation before fumigation but not total microbial biomass determined by microscopy.The ratio of CO₂ evolved-to-N mineralized followed fumigation changed in direct proportion to the ratio of fungal-to-bacterial biomass present in the soil before fumigation. Although more experiments with different soils should be performed, these results indicate that CO₂ evolved or N mineralized varies with the ratio of fungal-to-bacterial biomass initially present.     

Carbon mineralization in an organic soil, with and without added grass litter, from a high-CO2 environment at a carbon dioxide spring

Both CO₂-C production and the decomposability of grass leaf litter in a gley soil from a naturally occurring CO₂ spring were previously shown to be influenced by the atmospheric CO₂ concentrations under which the soil and litter were sampled. Here we investigate C mineralization in an organic soil from very high CO₂ environments (range 1220-3900 μl l⁻¹) at the same spring, and the effect of added leaf litter on CO₂-C production. Carbon mineralization in the organic soil was unusual in two respects: (1) the proportion of labile components was very high, with more than 11% of the initial soil C being metabolized to CO₂-C after 56 d at 25 °C; (2) rates of CO₂-C production in autumn samples increased on incubation, after an initial decline. Decomposition was initially more rapid in C3 Holcus lanatus (Yorkshire fog) than in C4 Pennisetum clandestinum (kikuyu) litter, but differed little in samples from different atmospheric CO₂ concentrations. Overall, the effects of environmental variables on estimates of litter decomposability in the organic soil were similar to, although much less marked than, those in the gley soil. Results suggest that organic components in the organic soil were metabolized at least as readily as those of added litter during the later stages of the 56-d incubation.     

Physical factors influencing decomposition of organic materials in soil aggregates

Glucose or starch labelled with ¹⁴C was mixed thoroughly into slurried soils. Aggregates of different sizes were obtained from the soils as they dried. The labelled substrates were considered to be distributed in both micro- and macropores in the aggregates. Control samples (labelled substrates in macropores only) were prepared by adding the labelled carbohydrates after the formation of the aggregates. The various samples were sterilized by γ-irradiation and stored at ?15°C.Samples were wetted to about ?20kPa, inoculated with soil organisms, and incubated for 4 weeks at 28°C in closed systems, which enabled regular measurement of ¹⁴CO₂ released.Based on the ¹⁴CO₂ released, it was concluded that starch was protected from microbial attack when present in micropores in aggregates made from fine sandy loam.After incubation samples were dried and rewetted. The flush of ¹⁴CO₂ released was twice as big for samples containing labelled starch compared with glucose, showing that disruption of aggregates, containing residual starch, and rearrangements of soil components are as important as chemical and biological factors in causing the flush of CO₂ resulting from wetting a soil. Mechanical disruption of the aggregates resulted in a similar flush of ¹⁴CO₂.     

Fluctuations in microbial biomass indices at different sampling times in soils from tussock grasslands

Microbial biomass in four topsoils from New Zealand tussock grasslands was estimated by three biochemical procedures at five sampling times over a 15 month period. In Conroy, Cluden and Tima soils, biomass C content was high in two sets of March (summer-autumn) samples and low in October (early spring) samples; in Carrick soil from a wetter, cooler environment, it was similar at all sampling times. Significant time-of-sampling variations occurred with Min-N flush in Tima and Carrick soils, and with adenosine 5'-triphosphate (ATP) content in three of the soils. Generally, the ratios of these biomass indices also varied significantly at some sampling times. Because of this variability, common factors could not validly be used with these soils for estimating biomass C contents from Min-flush or ATP values.The contribution of bacteria and fungi to the respiratory activity of the microbial biomass was unsuccessfully investigated using streptomycin and actidione as differential inhibitors of anabolic metabolism in the presence of added glucose. In three of the soils, rates of O₂ uptake did not generally increase significantly during incubation, even with added N, P, K and S or prior incubation overnight. In Conroy soil, rates did increase significantly, but the effects of the antibiotics separately and together could not be satisfactorily balanced.     

Limitations when quantifying microbial carbon and nitrogen by fumigation‐extraction in rooted soils

Soil microbial C and N (C_mic, N_mic) estimation by the chloroform fumigation‐extraction method is erroneous in densely rooted soils due to CHCl₃‐labile C and N compounds. The effect of a pre‐extraction with 50 mM K₂SO₄ and a pre‐incubation (conditioning at 25 °C for 7 days) on the flush in extractable, CHCl₃‐labile C (C‐flush) and N (N‐flush) was tested with reference to rooting density (0.3—75 mg root dry matter g^—1) in one arable and 3 grassland soils. In the arable soil and in the second horizon (10—20 cm) of a grassland soil, C‐flush values were not affected by the pre‐extraction. However, the pre‐extraction considerably reduced C‐flush values in the top soils of the grassland (above 10 cm). Only about 42 % was found in the pre‐extracted roots and the rest was lost during the pre‐extraction. The estimated concentrations of N_mic decreased due to pre‐extraction of soil samples with low root biomass. Clearly, the concentrations of N_mic were underestimated by introducing the pre‐extraction. Soil pre‐incubation reduced C‐flush values only slightly, whereas N‐flush values were not affected. It can be concluded that (1) CHCl₃‐labile root C and N is partly extracted with K₂SO₄ after pre‐incubation and (2) CHCl₃‐labile C and N removed with the roots during pre‐extraction is partly derived from microbial biomass. Soils with low rooting density (arable soils, grassland soils below approximately 10 cm depth) should therefore be fumigated and extracted without pre‐extraction. In densely rooted soils, fumigation extraction with and without pre‐extraction probably gives estimates for the minimum and maximum of C_mic and N_mic.     

Long-term fertilizer management in NERICA cultivated upland affects on soil bio-chemical properties

ABSTRACT

This study aims to characterize soil chemical properties and microbial biomass, greenhouse gas production, and organic matter dynamics in upland rice field as affected by the long-term fertilizer managements in Uganda. Soil total C (TC) and N (TN) contents were in the relatively smaller range under different fertilizer treatments, even after 20 crop seasons. However, available phosphate contents showed positive correlation with average yield of upland rice. Incubation experiments were conducted under aerobic or under flooding conditions to measure CO₂, methane, and nitrous oxide productions. After the incubation, soil samples were extracted to quantify nitrification rate for aerobic condition and ammonification rate for flooding condition. Soil microbial biomass carbon (MBC) and nitrogen were measured. Stable isotope ratio of ¹³C and ¹⁵N were also determined for the soil samples. CO₂ production potential under aerobic condition was higher than the flooding condition. The qCO₂ (CO₂/MBC) in the treatment applied with compost tended to be higher than the other treatments. Positive correlation between nitrous oxide production and nitrification was found. The delta ¹³C values of the soil samples indicated that the effect of C4 plants before rice cultivation still remained, while the contribution of biological N₂ fixation was little according to delta ¹⁵N values. These results indicate that soil microbial biomass in upland rice field of the long-term fertilizer experiment in Uganda was characterized with higher qCO₂. Greenhouse gas production was affected by fertilizer management, while soil organic C before the long-term experiment still remained in the experiment.     

Correlation between four methods to estimate total microbial biomass in stored,air-dried and glucose-amended soils

Correlation between the microbial volume, chloroform fumigation (CO₂-C flush), substrateinduced respiration (SIR) and ATP content methods to estimate microbial biomass was assessed on three New Zealand soils (two grassland, one arable) under three different treatments (stored, air-dried and glucose-amended). There were significant, positive correlations between all methods, r = 0.69–0.88, which were improved, r = 0.71–0.96, if the data for air-dried or glucose-amended soils were excluded from the analyses. The best agreement was between CO₂-C flush and ATP and the worst between CO₂-C flush and microbial volume. Exclusion of air-dried soil data improved these correlations.Estimates of microbial biomass for each soil often differed significantly between the four methods, when conversion factors cited in the literature were used. Ratios (i.e. conversion factors) between CO₂-C flush and ATP or SIR, or SIR and volume, were different to those cited in the literature, and only similar if specific data were excluded.We recommend that a minimum of two and preferably three methods be used to quantify the microbial population of soil, and that emphasis should be placed on the relative differences within and between soils using data which have not been converted to biomass C. Conversion of data to biomass C may result in substantial errors.     

The proportional mineralisation of microbial biomass and organic matter caused by air-drying and rewetting of a grassland soil

During the first few days after rewetting of an air-dried soil (AD-RW), microbial activity increases compared to that in the original moist soil, causing increased mineralisation (a flush) of soil organic carbon (C) and other nutrients. The AD-RW flush is believed to be derived from the enhanced mineralisation of both non-biomass soil organic matter (due to its physical release and enhanced availability) and microbial biomass killed during drying and rewetting. Our aim was to determine the effects of AD-RW on the mineralisation of soil organic matter and microbial biomass during and after repeated AD-RW cycles and to quantify their proportions in the CO₂-C flushes that resulted. To do this, a UK grassland soil was amended with ¹⁴C-labelled glucose to label the biomass and then given five AD-RW cycles, each followed by 7 d incubation at 25 °C and 50% water holding capacity. Each AD-RW cycle increased the amount of CO₂-C evolved (varying from 83 to 240 μg g⁻¹ soil), compared to the control with, overall, less CO₂-C being evolved as the number of AD-RW cycles increased. In the first cycle, the amount of biomass C decreased by 44% and microbial ATP by 70% while concentrations of extractable C nearly doubled. However, all rapidly recovered and within 1.3 d after rewetting, biomass C was 87% and ATP was 78% of the initial concentrations measured prior to air-drying. Similarly, by 2 d, extractable organic C had decreased to a similar concentration to the original. After the five AD-RW cycles, the amounts of total and ¹⁴C-labelled biomass C remaining in the soil accounted for 60 and 40% of those in the similarly incubated control soil, respectively. Soil biomass ATP concentrations following the first AD-RW cycle remained remarkably constant (ranging from about 10 to 14 μmol ATP g⁻¹ biomass C) and very similar to the concentration in the fresh soil prior to air-drying. We developed a simple mathematical procedure to estimate the proportion of CO₂-C derived from biomass C and non-biomass C during AD-RW. From it, we estimate that, over the five AD-RW cycles, about 60% of the CO₂-C evolved came from mineralisation of non-biomass organic C and the remainder from the biomass C itself.     

Nitrous oxide production at different soil moisture contents in an arable soil in China

Abstract

Laboratory incubations were conducted to investigate nitrous oxide (N₂O) production from a subtropical arable soil (Typic Plinthodults) incubated at different soil moisture contents (SMC) and with different nitrogen sources using a 10% (v/v) acetylene (C₂H₂) inhibitory technique at 25°C. The production of N₂O and CO₂ was monitored during the incubations and changes in the contents of KCl-extractable NO^? ₃-N and NH⁺ ₄-N were determined. The production of N₂O increased slightly with an increase in SMC from 40% water-holding capacity (WHC) to 70% WHC, but increased dramatically at 100% WHC. After incubation the NO^? ₃-N content increased even at a SMC of 100% WHC. At a SMC of 100% WHC, the addition of NH⁺ ₄-N promoted the production of N₂O and CO₂, whereas the addition of NO^? ₃-N decreased N₂O production. Compared with the incubation without C₂H₂, the presence of C₂H₂ increased NH⁺ ₄-N content, but decreased NO^? ₃-N content, and there was no significant difference in N₂O production. These results indicate that heterotrophic nitrification contributes to N₂O production in the soil.     

Bacterial community structure in fumigated soil

Soil microbial biomass has been determined since the mid 1970's by the chloroform fumigation incubation technique as proposed by Jenkinson and Powlson (1976). The microbial biomass C can be determined by subtracting the CO₂ emitted from an unfumigated soil (mineralization of soil organic matter) from that emitted from a chloroform fumigated inoculated soil (mineralization of soil organic matter and killed soil microorganisms) and dividing the difference by a proportionality factor (k_C = 0.45). The question remained which microorganisms recolonized a fumigated soil. An arable soil was fumigated for one day with ethanol-free chloroform or left unfumigated and incubated aerobically after removal of the chloroform for 10 days. The bacterial population structures were determined in the fumigated and unfumigated soil after 0, 1, 5 and 10 days by means of 454 pyrosequencing of the 16S rRNA gene. Fumigating the arable soil reduced significantly the relative abundance of phylotypes belonging to different groups, but increased the relative abundance of only four genera belonging to two phyla (Actinobacteria and Firmicutes) and two orders (Actinomycetales and Bacillales). The relative abundance of phylotypes belonging to the Micromonospora (Micromonosporaceae) increased significantly from 0.2% in the unfumigated soil to 6.7% in the fumigated soil and that of Bacillus (Bacillaceae) from 3.6% to 40.8%, Cohnella (Paenibacillaceae) from undetectable amounts to 0.6% and Paenibacillus (Paenibacillaceae) from 0.3% to 4.2%. The relative percentage of phylotypes belonging to the Acidobacteria, Bacteroidetes, Chloroflexi, Gemmatimonadetes and Proteobacteria (α- β-, δ- and γ-Proteobacteria) were significantly lower in the fumigated than in the unfumigated soil and in most of them the relative abundance of different bacterial orders (i.e. Gp3, Gp4, Gp6, Sphingobacteriales, Gemmatimonadales, Rhodospirillales, Burkholderiales, Xanthomonadales) was reduced strongly (P < 0.001). It was found that the relative abundance of a wide range of bacteria was reduced shortly after fumigating an arable soil, but only a limited group of bacteria increased in a fumigated arable soil indicating a capacity to metabolize the killed soil microorganisms or recolonize a fumigated soil.     

Response of microbial biomass to alternate moist and dry conditions in a soil incubated with 14C- and 15N-labelled plant material

A red mediterranean soil was incubated for 1.5 yr, with ¹⁴C- and ¹⁵N-labelled plant material under constant temperature and moisture conditions. Then a portion of the soil was submitted to 4 rapid drying (at 40°C) and rewetting cycles. The duration of the dry periods ranged from 8 to 10 days and the wet periods from 15 to 20 days. Another portion of the soil was incubated under continuously moist conditions. At the end of each dry and moist period, biomass-C and -N were estimated, using the chloroform fumigation technique. The portion of biomass killed on drying and that restored after rewetting were calculated, by the difference between the sizes of biomass present after the dry and the moist periods.Soil drying destroyed $\text{1}\text{3}$ to $\text{1}\text{4}$ of biomass and at each cycle, after remoistening, the biomass was progressively restored to approximately the same size as before drying.The labelled-C to total-C ratio of the CO₂ released from undisturbed and continuously moist soil, ranged from 6 to 7%. In biomass, which survived the drying, the values ranged from 20 to 22%, whereas in the killed biomass they ranged between 7 and 8%, i.e. the same orders of magnitude as that of CO₂ evolved from undisturbed soil.A comparison of the labelled-C to total-C ratio of (1) CO₂C released from undisturbed and continuously moist soil, (2) the extra CO₂-C evolved as a result of alternate drying-remoistening conditions, (3) CO₂C released from the soil at the end of moist periods, (4) the C of microbial biomass which survived the drying, and (5) the C of the biomass present at the end of the moist periods, revealed that the calculated portion of biomass killed on drying essentially corresponded to a still relatively “active” fraction of biomass and that the biomass surviving rapid drying, essentially corresponded to a dormant and protected fraction.In contrast to the labelled-C to total-C ratio, the labelled-N to total-N ratio, in the fraction of biomass which was destroyed on drying, was not different from that of the surviving fraction. During incubation, labelled nitrates accumulated progressively in soil; transformations of N were probably affected by a remetabolization of the nitrates and of material made decomposable on drying, including destroyed microflora and non-biomass material.     

Dynamics of maize (Zea mays L.) leaf straw mineralization as affected by the presence of soil and the availability of nitrogen

An incubation experiment was carried out with maize (Zea mays L.) leaf straw to analyze the effects of mixing the residues with soil and N amendment on the decomposition process. In order to distinguish between soil effects and nitrogen effects for both the phyllospheric microorganisms already present on the surface of maize straw and soil microorganisms the N amendment was applied in two different placements: directly to the straw or to the soil. The experiment was performed in dynamic, automated microcosms for 22 days at 15 °C with 7 treatments: (1) untreated soil, (2) non-amended maize leaf straw without soil, (3) N amended maize leaf straw without soil, (4) soil mixed with maize leaf straw, (5) N amended soil, (6) N amended soil mixed with maize leaf straw, and (7) soil mixed with N amended maize leaf straw. ¹⁵NH₄¹⁵NO₃ (5 at%) was added. Gas emissions (CO₂, ¹³CO₂ and N₂O) were continuously recorded throughout the experiment. Microbial biomass C, biomass N, ergosterol, δ¹³C of soil organic C and of microbial biomass C as well as ¹⁵N in soil total N, mineral N and microbial biomass N were determined in soil samples at the end of the incubation. The CO₂ evolution rate showed a lag-phase of two days in the non-amended maize leaf straw treatment without soil, which was completely eliminated when mineral N was added. The addition of N generally increased the CO₂ evolution rate during the initial stages of maize leaf straw decomposition, but not the cumulative CO₂ production. The presence of soil caused roughly a 50% increase in cumulative CO₂ production within 22 days in the maize straw treatments due to a slower decrease of CO₂ evolution after the initial activity peak. Since there are no limitations of water or N, we suggest that soil provides a microbial community ensuring an effective succession of straw decomposing microorganisms. In the treatments where maize and soil was mixed, 75% of microbial biomass C was derived from maize. We concluded that this high contribution of maize using microbiota indicates a strong influence of organisms of phyllospheric origin to the microbial community in the soil after plant residues enter the soil.