Microbial response of soils with organic and conventional management history to the cultivation of Bacillus thuringiensis (Bt)-maize under climate chamber conditions

An experiment was carried out in a climate chamber to analyse if Bt-maize may cause particular changes in soils with different levels of microbial biomass and activity due to long-term management history. Among the soils selected, the ones managed organically for 30 years exhibited twice the microbial biomass and 2.6 times the dehydrogenase activity (DHA) of the soil from a field with long-term conventional maize monoculture. Soils were cultivated twice in a row with Bt-maize, its near-isogenic line and a conventional breeding line. We tested the hypotheses that (a) soil microbial biomass and activity are affected by the cultivation of Bt-maize and that (b) the influence of Bt-maize depends on the level of soil microbial biomass and activity. Shoot and root yield and shoot C-content of Bt-maize were higher than the ones of the near-isogenic line. DHA under Bt-maize was 6 % higher, and the metabolic quotient for CO₂ (qCO₂) was 9 % lower than under its near-isogenic line, giving some support to hypothesis (a). No significant interactions of the soils and the varieties used were found in this study, thus hypothesis (b) was not confirmed, and soils with different microbial biomass and activity appear to react in a similar way to the cultivation of Bt-maize.     

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.     

Respiration pattern and microbial use of field-grown transgenic Bt-maize residues

A 49-day incubation experiment was carried out with the addition of field-grown maize stem and leaf residues to soil at three different temperatures (5, 15, and 25 °C). The aim was to study the effects of two transgenic Bt-maize varieties in comparison to their two parental non-Bt varieties on the mineralization of the residues, on their incorporation into the microbial biomass and on changes in the microbial community structure. The stem and leaf residues of Novelis-Bt contained 3.9 μg g⁻¹ dry weight of the Bt toxin Cry1Ab and those of Valmont-Bt only 0.8 μg g⁻¹. The residues of the two parental non-Bt varieties Nobilis and Prelude contained higher concentrations of ergosterol (+220%) and glucosamine (+190%) and had a larger fungal C-to-bacterial C ratio (+240%) than the two Bt varieties. After adding the Bt residues, an initial peak in respiration of an extra 700 μg CO₂-C g⁻¹ soil or 4% of the added amount was observed in comparison to the two non-Bt varieties at all three temperatures. On average of the four varieties, 19-38% of the maize C added was mineralized during the 49-day incubation at the three different temperatures. The overall mean increase in total maize-derived CO₂ evolution corresponded to a Q₁₀ value of 1.4 for both temperature steps, i.e. from 5 to 15 °C and from 15 to 25 °C. The addition of maize residues led to a strong increase in all microbial properties analyzed. The highest contents were always measured at 5 °C and the lowest at 25 °C. The variety-specific contents of microbial biomass C, biomass N, ATP and adenylates increased in the order Novelis-Bt ? Prelude<Valmont-Bt ? Nobilis. The mineralization of Novelis-Bt residues with the highest Bt concentration and lowest N concentration and their incorporation into the microbial biomass was significantly reduced compared to the parental non-Bt variety Nobilis. These negative effects increased considerably from 5 to 25 °C. The transgenic Bt variety Valmont did not show further significant effects except for the initial peak in respiration at any temperature.     

Transgenic Bt plants decompose less in soil than non-Bt plants

Bt plants are plants that have been genetically modified to express the insecticidal proteins (e.g. Cry1Ab, Cry1Ac, Cry3A) from subspecies of the bacterium, Bacillus thuringiensis (Bt), to kill lepidopteran pests that feed on corn, rice, tobacco, canola, and cotton and coleopteran pests that feed on potato. The biomass of these transgenic Bt plants (Bt+) was decomposed less in soil than the biomass of their near-isogenic non-Bt plant counterparts (Bt−). Soil was amended with 0.5, 1, or 2% (wt wt⁻¹) ground, dried (50 °C) leaves or stems of Bt corn plants; with 0.5% (wt wt⁻¹) ground, dried biomass of Bt rice, tobacco, canola, cotton, and potato plants; with biomass of the near-isogenic plants without the respective cry genes; or not amended. The gross metabolic activity of the soil was determined by CO₂ evolution. The amounts of C evolved as CO₂ were significantly lower from soil microcosms amended with biomass of Bt plants than of non-Bt plants. This difference occurred with stems and leaves from two hybrids of Bt corn, one of which had a higher C:N ratio than its near-isogenic non-Bt counterpart and the other which had essentially the same C:N ratio, even when glucose, nitrogen (NH₄NO₃), or glucose plus nitrogen were added with the biomass. The C:N ratios of the other Bt plants (including two other hybrids of Bt corn) and their near-isogenic non-Bt counterparts were also not related to their relative biodegradation. Bt corn had a significantly higher lignin content than near-isogenic non-Bt corn. However, the lignin content of the other Bt plants, which was significantly lower than that of both Bt and non-Bt corn, was generally not statistically significantly different, although 10-66% higher, from that of their respective non-Bt near-isolines. The numbers of culturable bacteria and fungi and the activity of representative enzymes involved in the degradation of plant biomass were not significantly different between soil amended with biomass of Bt or non-Bt corn. The degradation of the biomass of all Bt plants in the absence of soil but inoculated with a microbial suspension from the same soil was also significantly less than that of their respective inoculated non-Bt plants. The addition of streptomycin, cycloheximide, or both to the soil suspension did not alter the relative degradation of Bt+ and Bt− biomass, suggesting that differences in the soil microbiota were not responsible for the differential decomposition of Bt+ and Bt− biomass. All samples of soil amended with biomass of Bt plants were immunologically positive for the respective Cry proteins and toxic to the larvae of the tobacco hornworm (Manduca sexta), which was used as a representative lepidopteran in insect bioassays (no insecticidal assay was done for the Cry3A protein from potato). The ecological and environmental relevance of these findings is not clear.     

Persistence of Cry1Ac Protein from Transgenic Bt Cotton Cultivation and Residue Returning in Fields and Its Effect on Functional Diversity of Soil Microbial Communities

The persistence of Cry1Ac protein in the soil and its effect on soil microbial communities are a core issue in assessing the ecological risk of transgenic Bacillus thuringiensis (Bt) cotton. In this study a field experiment was conducted on the cultivation of transgenic Bt cotton (Jin 26 and BtJi 668) with the immediate returning of residues to the fields, in order to quantify the Cry1Ac protein content in the fields and investigate its effects on the functional diversity of soil microbial communities. Cry1Ac protein in the residue-soil mixture was gradually degraded in the transgenic Bt cotton fields. After transgenic Bt cotton straw was returned to the fields for 30 d, 63.73% and 58.33% of the initial amounts of Cry1Ac protein were degraded in the Jin 26 and BtJi 668 fields, respectively. Before the crops were sown in the following year (180 d after returning the straw), no Cry1Ac protein was detected in the fields. After returning the cotton straw to the fields for 30 d, the Shannon-Wiener and McIntosh indices of soil microbial communities in the transgenic Bt cotton fields were significantly higher than those in the non-transgenic cotton fields. Meanwhile, the utilization of carbon sources including amino acids, amines, and carbohydrates by the soil microbial communities significantly increased. Both the McIntosh index and the utilization of carbohydrates increased until 180 d. Principal component analysis revealed that amino acids, amides, and carbohydrates were the main carbon sources distinguishing the two principal component factors. These findings indicated that Cry1Ac protein did not accumulate in the fields after transgenic Bt cotton was planted for one year and the residues were immediately returned to the fields; however, the original functional diversity of soil microbial communities was affected continuously.     

The adenylate energy charge of the soil microbial biomass

A method was developed for measuring adenosine 5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP) and adenosine 5'-monophosphate (AMP) in soil. All three adenine nucleotides were extracted from soil with a solution of trichloroacetic acid, paraquat and phosphate. ATP was measured in the neutralised (pH 7.4) soil extracts by the fire-fly luciferin-luciferase system. ADP was measured as ATP after incubating the neutralised extracts with pyruvate kinase (PK) and phosphoenolpyruvate (PEP) to convert ADP to ATP. AMP was converted to ATP by incubation with the coupled PK-PEP-myokinase system and measured as ATP. The quantities of nucleotides present in the extracts were corrected for incomplete extraction from soil by measuring the percentage recovery of added ATP, ADP and AMP. The adenylate energy charge (AEC) was calculated from the formula AEC = [[ATP] + 0.5[ADP]]/[[ATP] + [ADP] + [AMP]]. Measurements were made on (1) fresh soil, extracted as soon as possible after field sampling (2) soil stored air-dry at 5°C for 18 days and (3) soil stored air-dry at 5°C for 57 days and then rewetted to the original field moisture content and incubated aerobically for 2.5 h at 10°C before extraction.In moist soil the biomass maintains both ATP and AEC at levels close to those of activity growing cells, even though little of the biomass in soil can be in active growth at any given time. ATP accounted for 77% of the total adenine nucleotides (A_T) in the fresh soil, with an AEC of 0.85 (a value comparable to that found in microorganisms undergoing active growth in vitro. In contrast, ATP only accounted for 28% of A_T in the air-dried soil, with an AEC of 0.46. When the air-dried soil was rewetted, ATP increased to 66% of A_T and the AEC increased to 0.76. However, A_T in the air-dried soil (7.65 nmol g^?1 soil) was of the same order as that in rewetted soil (6.70 nmol g^?1) even though the AEC's were very different.These results show that the soil microbial biomass does not maintain a high AEC when air-dried. Once remoistened, the population tends to restore its AEC to the original value. This restoration occurs so rapidly that it cannot be due to the formation of a new biomass.     

Microbial biomass dynamics in recently air-dried and rewetted soils compared to others stored air-dry for up to 103 years

Our aim was to compare the soil microbial biomass concentration and its activity (measured as CO₂-C evolved) following the rewetting and aerobic incubation of soils which have previously been stored air-dry for different periods. Some of the soils have been stored in the Rothamsted sample archive for 103 years, others were comparable freshly sampled soils following air-drying and rewetting and other soils were stored air-dry for 2 years then rewetted for the work described here. Following air-drying, soil ATP concentrations were variable in recently air-dried soil, comprising about 10-35% of the initial ATP concentrations in fresh soil. Following rewetting, the percentage recovery of ATP increased in all soils by 7 days, then declined to between 73% and 87% of the original ATP concentration in the air-dried soils by day 12. Storage of air-dried soils decreased the ability of the microbial biomass to restore its ATP concentrations. For example, the ATP concentration in a soil sampled from stubbed (i.e. tree seedling, saplings and bushes cut frequently to ground level) grassland of the Broadbalk continuous wheat experiment at Rothamsted then air-dried for 2 years was only about 14% of that in the fresh soil at 2 days after rewetting. In other soils from the Hoosfield Barley Experiment, also at Rothamsted, previously given NPK or FYM since 1852, and sampled then stored air-dry for between 13 and 83 years, from 52% to 57% of the ATP in the comparable fresh soils was measured at two days after rewetting. The soil ATP concentration then changed little more up to 12 days. One of the most interesting findings was that while the microbial biomass ATP concentration in the above NPK soils only ranged from about 2 to 4 μmol ATP g⁻¹ biomass C, in the FYM soil the microbial biomass ATP concentrations (range 11.5-13.6 μmol ATP g⁻¹ biomass C) were the same as we repeatedly measure in fresh moist aerobic soil. We do not yet know the reasons for this. More than twice as much CO₂-C was evolved from the long-term stored soils than from freshly sampled ones. However, the specific respiration of the microbial biomass did not change much after the first 12 years of storage, indicating that loss of viability mainly occurred in the earlier years.     

Adenylates as an estimate of microbial biomass C in different soil groups

Adenylate (i.e. adenosine tri- (ATP), di- (ADP) and monophosphates (AMP)) and microbial biomass C data were collected over a wide range of sites including forest floor layers and forest, grassland and arable soils. Microbial biomass C was measured by fumigation extraction and adenylates after alkaline Na₃PO₄/DMSO/EDTA extraction and HPLC detection. Our aims were (1) to test whether the sum of adenylates is a better estimate for microbial biomass than the determination of ATP, (2) to compare our conversion values with those proposed by others, and (3) to analyse whether soil properties or land use form affect the relationships between ATP, adenylates and microbial biomass C. A close relationship was found between microbial biomass C and ATP (r=0.96), but also with the sum of adenylates (r=0.96) within all appropriately conditioned soil samples (n=112). In the mineral soil (n=98), the geometric means of the ATP-to-microbial biomass C ratio and the adenylates-to-microbial biomass C ratio were 7.4 and 11.4 μmol g⁻¹, respectively. The mean ratios did not differ significantly between the different texture classes and land use forms. In the forest floor, the ATP-to-microbial biomass C ratio and the adenylates-to-microbial biomass C ratio were both roughly two-thirds of those of the mineral soil. The average adenylate energy charge (AEC) of all soil samples was 0.79 and showed a strong negative relationship with the soil pH (r=−0.69). However, the AEC is presumably only indirectly affected by the soil pH.     

Adenylate energy charge measurements in soil

Adenosine 5'-triphosphate (ATP), adenosine 5'-diphosphatc (ADP) and adenosine 5'-monophosphate (AMP) were extracted from soil with either a solution of trichloroacetic acid, paraquat and phosphate (TCA reagent) or a mixture of chloroform, sodium hydrogen carbonate, phosphate and adenosine (NaHCO₃ reagent). Standard enzymic procedures were used to convert ADP and AMP to ATP, which was measured by the fire-fly luciferin-luciferase system. The measured quantities of nucleotides were corrected for incomplete extraction using the percentage recoveries of added ATP, ADP and AMP. The adenylate energy charge ratio (AEC) was calculated from the formula AEC = ([ATP] + 0.5 [ADP])/([ATP] + [ADP] + [AMP]).Measurements were made on a grassland soil, following a conditioning incubation at 15°C and 50% WHC for 7 days. Additional measurements were made on the same soil after a further 50- or 100-day incubation at 25°C and 50% WHC, with or without an amendment of 1100 μg ryegrass Cg⁻¹ soil, added at the end of the conditioning incubation. Biomass-ATP concentration, measured in TCA extracts, changed little, even on prolonged incubation, and was maintained at a level comparable to that observed in earlier work (about 10 p mol ATP g⁻¹ biomass C). AEC values in TCA soil extracts were high (0.8–0.9) for all soil treatments and independent of substrate addition or length of incubation.In contrast, AEC was low (0.4) in fresh soil extracted with NaHCO₃ reagent, but increased to 0.6 when ryegrass was incubated with the soil for 50 days. Although the total adenine nucleotide pool (i.e. [ATP] + [ADP] + [AMP]) was similar as measured in NaHCO₃ and in TCA soil extracts, both energy charge and ATP content were lower in the NaHCO₃ extracts. It was therefore concluded that the main reason for the lower AECs observed with the NaHCO₃ reagent was that microbial ATPases were still active during extraction and caused appreciable hydrolysis of microbial ATP to ADP and AMP. In contrast, the TCA reagent rapidly inactivates ATPases and is therefore preferable for extracting adenine nucleotides from soil.The results indicate that the soil microbial biomass, although a mainly dormant population, maintains both AEC and ATP at levels characteristic of exponentially growing organisms in vitro, even during prolonged incubation without fresh substrate. It was also concluded that roots make a negligible contribution to total ATP extracted from fresh sieved soil.     

Decomposition of pea and maize straw in Pakistani soils along a gradient in salinity

Maize straw and pea straw were added to five Pakistani soils from a gradient in salinity to test the following hypotheses: Increasing salinity at high pH decreases proportionally (1) the decomposition of added straw and (2) the resulting net increase in microbial biomass. In the non-amended control soils, salinity had depressive effects on microbial biomass C, biomass N, but not on biomass P and ergosterol. The ratios microbial biomass C-to-N and biomass C-to-P decreased consistently with increasing salinity. In contrast, the ergosterol-to-microbial biomass C ratio was constant in the four soils at pH>8.9, but nearly doubled in the most saline, but least alkaline, soil (pH 8.2). The addition of the maize and pea straw always increased the contents of microbial biomass C, biomass N, biomass P and ergosterol, but without clear effects of salinity. Highest mean contents of microbial biomass C and biomass N were measured at day 0, immediately after the straw was added. Straw amendments increased the CO₂ evolution rates of all five soils without any effect of salinity. The same was true for total C and total N in the two fractions of particulate organic matter (POM) 63–400 μm and >400 μm. Lowest percentage of straw-derived CO₂-C and highest recoveries of POM-C and POM-N were observed in the maize straw treatment and the reverse in the pea straw treatment. Yield coefficients were calculated for maize and pea straw based on the assumption that the balance gap between CO₂ and the amount of POM can be fully assigned to microbial products.     

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.     

Exposure and effects assessments of Bt-maize on non-target organisms (gastropods, microarthropods, mycorrhizal fungi) in microcosms

Potential differences between Bt-maize (MEB307 expressing the insecticidal Cry1Ab protein) and a near-isogenic non-Bt variety (Monumental) in their influence on the garden snail (Helix aspersa), soil microarthropods (Collembola, Actinedida, Acaridida, Gamasida and Oribatida) and mycorrhizal fungi were studied. Growing snails were caged in microcosms allowing the development of Bt or non-Bt-maize (Zea mays L.) on a sandy loam soil. After 3 months exposure, survival and growth of snails were similar in both treatments. Cry1Ab protein was detected in the Bt-maize leaves (22–42.2 μg Bt protein g⁻¹ dry wt), in the snail tissues (0.04–0.11 μg Bt-protein g⁻¹ dry wt) and in their faeces (0.034–5 μg Bt-protein g⁻¹ dry wt). Total soil microarthropod abundance and diversity were similar between control (non-Bt-maize) and the genetically modified (GM) Bt-maize microcosms. The mycorrhizal colonization of roots did not differ between Bt and non-Bt-maize (frequency of mycorrhizal roots was 88.7% and 83.3% respectively). The mycorrhizal infectivity of soils, expressed as MI₅₀ (minimum soil dry weight required to colonize 50% of plants) was measured using red clover. MI₅₀ was similar for soils where Bt or non-Bt-maize was cultivated for 4 months. The detection of Cry1Ab protein in the viscera and faeces of H. aspersa exposed to Bt-maize indicates that snails contribute to the transfer of the Bt-protein from plant to soil or snail predators. This may constitute an alternative route of exposure for Bt-protein in soil, but this was without a negative influence on mycorrhizal fungi or microarthropods. Results showed that Bt-maize was not toxic for the selected non-target species exposed for 3 or 4 months. The microcosms and analyses used in this study represent new methods for assessing effects of chronic exposure to GM plants of several diverse, yet ecologically and temporally associated species. As the soil organisms we studied can also be used in standardized ecotoxicological tests (XP X31-205-2 for mycorrhizal fungi, ISO 11267 for Collembola and ISO 15952 for snails), microcosm exposures represent a way to link laboratory and field methods for the ecotoxicological evaluation of GM plants.     

Decomposition of maize residues after manipulation of colonization and its contribution to the soil microbial biomass

A 28-day incubation experiment at 12°C was carried out on the decomposition of maize leaf litter to answer the questions: (1) Is the decomposition process altered by chemical manipulations due to differences in the colonization of maize leaf litter? (2) Do organisms using this maize material contribute significantly to the soil microbial biomass? The extraction of the maize straw reduced its initial microbial biomass C content by 25%. Fumigation and extraction eliminated the microbial biomass by 88%. In total, 17% of added maize straw C was mineralized to CO₂ during the 28-day incubation at 12°C in the treatment with non-manipulated straw. Only 14% of added C was mineralized in the treatment with extracted straw as well as in the treatment with fumigated and extracted straw. The net increase in microbial biomass C was 79 μg g^?1 soil in the treatment with non-manipulated straw and an insignificant 9 μg g^?1 soil in the two treatments with manipulated straw. However, the net increase did not reflect the fact that the addition of maize straw replaced an identical 58% (≈180 μg g^?1 soil) of the autochthonous microbial biomass C₃-C in all three straw treatments. In the two treatments with manipulated straw, the formation of maize-derived microbial biomass C₄-C was significantly reduced by 25%. In the three straw treatments, the ratio of fungal ergosterol-to-microbial biomass C ratio showed a constant 60% increase compared to the control, and the contents of glucosamine and muramic acid increased by 18%. The average fungal C/bacterial C ratio was 3.6 in the soil and 5.0 in the recovered maize straw, indicating that fungal dominance was not altered by the initial chemical manipulations of the maize straw-colonizing microorganisms.     

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.     

Long‐term effects of leguminous cover crops on microbial indices and their relationships in soils of a coconut plantation of a humid tropical region

In this study, leguminous crops like Atylosia scarabaeoides, Centrosema pubescens, Calopogonium mucunoides, and Pueraria phaseoloides. grown as soil cover individually in the interspaces of a 19‐yr‐old coconut plantation in S. Andaman (India) were assessed for their influence on various microbial indices (microbial biomass C, biomass N, basal respiration, ergosterol, levels of ATP, AMP, ADP) in soils (0–50 cm) collected from these plots after 10 years. The effects of these cover crops on . CO₂ (metabolic quotient), adenylate energy charge (AEC), and the ratios of various soil microbial properties viz., biomass C : soil organic C, biomass C : N, biomass N : total N, ergosterol : biomass C, and ATP : biomass C were also examined. Cover cropping markedly enhanced the levels of organic matter and microbial activity in soils after the 10‐yr‐period. Microbial biomass C and N, basal respiration, . CO₂, ergosterol and levels of ATP, AMP, ADP in the cover‐cropped plots significantly exceeded the corresponding values in the control plot. While the biomass C : N ratio tended to decrease, the ratios of biomass N : total N, ergosterol : biomass C, and ATP : biomass C increased significantly due to cover cropping. Greater ergosterol : biomass C ratio in the cover‐cropped plots indicated a decomposition pathway dominated by fungi, and high . CO₂ levels in these plots indicated a decrease in substrate use efficiency probably due to the dominance of fungi. The AEC levels ranged from 0.80 to 0.83 in the cover‐cropped plots, thereby reflecting greater microbial proliferation and activity. The ratios of various microbial and chemical properties could be assigned to three different factors by principal components analysis. The first factor (PC1) with strong loadings of ATP : biomass C ratio, AEC, and . CO₂ reflected the specific metabolic activity of soil microbes. The ratios of ergosterol : biomass C, soil organic C : total N, and biomass N : total N formed the second factor (PC2) indicating a decomposition pathway dominated by fungi. The biomass C : N and biomass C : soil organic C ratios formed the third principal component (PC3), reflecting soil organic matter availability in relation to nutrient availability. Overall, the study suggested that Pueraria phaseoloides. or Atylosia scarabaeoides were better suited as cover crops for the humid tropics due to their positive contribution to soil organic C, N, and microbial activity.     

小麦和玉米秸秆腐解特点及对土壤中碳、氮含量的影响

通过室内模拟培养试验,揭示了不同水分条件下小麦和玉米秸秆在土壤中的腐解特点及对土壤碳、氮含量的影响。结果表明,1)水分条件对有机物质腐解的影响较大,在32 d的培养期间,相对含水量为60%(M60)时,土壤CO2释放速率始终低于含水量80%(M80)的处理。M60条件下释放的CO2-C量占秸秆腐解过程中释放碳总量的40.1%,而M80条件下达到51.5%;M60条件下,添加秸秆土壤中有机碳含量平均提高2.24 g/kg,显著高于M80条件下的1.43 g/kg。2)添加玉米秸秆的土壤,在培养期内CO2释放速率始终高于小麦秸秆处理,CO2-C累积释放量和有机碳净增量分别为408.35 mg/pot和2.12 g/kg;而小麦秸秆处理分别仅为378.94 mg/pot和1.56 g/kg,两种秸秆混合的处理介于二者之间。3)与未添加秸秆相比,土壤中添加小麦或玉米秸秆后,土壤有机碳、微生物量碳、全氮和微生物量氮含量均显著提高,且数量上总体趋势表现为:玉米秸秆两种秸秆混合小麦秸秆。可见,适宜水分条件有利于秸秆腐解过程中秸秆中碳向无机碳方向转化,而不利于向土壤有机碳方向转化;且玉米秸秆比小麦秸秆更易腐解。秸秆在土壤中腐解对补充土壤碳、氮作用很大,可改善土壤微生物生存条件,提高土壤质量。     

Criteria for measurement of microbial growth and activity in soil

Changes in CO₂ evolution, phosphatase and urease activity and ATP contents were related to bacterial and fungal biomass determined microscopically during glucose mineralization at different concentrations of mineral nutrients. Similar results were obtained in a sandy loam and a clay soil except that in the clay the increase in microbial and enzyme activities were delayed. Higher initial rates of CO₂ evolution were noted after the addition of P to a glucose and N amended soil at C:P ratios greater than 30:1. Increases in phosphatase activity coincided with increases in bacterial and fungal populations only in treatments without inorganic P. Peak rates of CO₂ evolution preceded biomass production by 18–24 h, therefore, CO₂ evolution rates did not show a correlation on normal regression analysis with biomass. Soil ATP content was influenced by P concentrations and soil type. ATP was therefore not a specific indicator of biomass in the detailed studies where P concentrations and sequential growth of bacteria and fungi were major factors. Soil urease increased with bacterial and fungal populations. It did not respond to P other than through microbial biomass and was highly correlated with microbial biomass. The results show that no one measurement of microbial biomass or activity is sufficient to interpret microbial growth in the soil system. Each of the criteria measured were sensitive to specific conditions affecting biomass and activity.     

Short-term effects of earthworm activity and straw amendment on the microbial C and N turnover in a remoistened arable soil after summer drought

Short-term effects of actively burrowing Octolasion lacteum (Örl.) (Lumbricidae) on the microbial C and N turnover in an arable soil with a high clay content were studied in a microcosm experiment throughout a 16 day incubation. Treatments with or without amendment of winter wheat straw were compared under conditions of a moistening period after summer drought. The use of ¹⁴C labeled straw allowed for analyzing the microbial use of different C components. Microbial biomass C, biomass N and ergosterol were only slightly affected by rewetting and not by O. lacteum in both cases. Increased values of soil microbial biomass were determined in the straw treatments even after 24 h of incubation. This extra biomass corresponded to the initial microbial colonization of the added straw. O. lacteum significantly increased CO₂ production from soil organic matter and from the ¹⁴C-labeled straw. Higher release rates of ¹⁴C-CO₂ were recorded shortly after insertion of earthworms. This effect remained until the end of the experiment. O. lacteum enhanced N mineralization. Earthworms significantly increased both mineral N content of soil and N leaching in the treatments without straw addition. Moreover, earthworms slightly reduced N immobilization in the treatments with straw addition. The immediate increase in microbial activity suggests that perturbation of soil is more important than substrate consumption for the effect of earthworms on C and N turnover in moistening periods after drought.     

Microbial biomass and activity indices after organic substrate addition to a selenium-contaminated soil

High concentrations of Se in soil might have negative effects on microorganisms. For this reason, the effect of organic substrate addition (glucose + maize straw) on Se volatilisation in relation to changes in microbial biomass and activity indices was investigated using an artificially Se-contaminated soil. Microbial biomass N was reduced on average by more than 50% after substrate addition, but adenylate energy charge (AEC) and metabolic quotient qCO₂ were both increased. The Se content decreased by nearly 30% only with the addition of the organic substrate at 25°C. No significant Se loss occurred without substrate at 25°C or with substrate at 5°C. In the two treatments with substrate addition, the substrate-derived CO₂ evolution was about 30% lower with Se addition than without. In contrast, Se had no effect on any of the other soil microbial indices analysed, i.e. microbial biomass C, microbial biomass N, adenosine triphosphate (ATP), AEC, ATP-to-microbial biomass C, and qCO₂.     

Variations in soil microbial biomass and crop roots due to differing resource quality inputs in a tropical dryland agroecosystem

The influence of exogenous organic inputs on soil microbial biomass dynamics and crop root biomass was studied through two annual cycles in rice-barley rotation in a tropical dryland agroecosystem. The treatments involved addition of equivalent amount of N (80 kg N ha⁻¹) through chemical fertilizer and three organic inputs at the beginning of each annual cycle: Sesbania shoot (high-quality resource, C:N 16, lignin:N 3.2, polyphenol+lignin:N 4.2), wheat straw (low-quality resource, C:N 82, lignin:N 34.8, polyphenol+lignin:N 36.8) and Sesbania+wheat straw (high-and low-quality resources combined), besides control. The decomposition rates of various inputs and crop roots were determined in field conditions by mass loss method. Sesbania (decay constant, k=0.028) decomposed much faster than wheat straw (k=0.0025); decomposition rate of Sesbania+wheat straw was twice as fast compared to wheat straw. On average, soil microbial biomass levels were: rice period, Sesbania?Sesbania+wheat straw>wheat straw?fertilizer; barley period, Sesbania+wheat straw>Sesbania?wheat straw?fertilizer; summer fallow, Sesbania+wheat straw>Sesbania>wheat straw?fertilizer. Soil microbial biomass increased through rice and barley crop periods to summer fallow; however, in Sesbania shoot application a strong peak was obtained during rice crop period. In both crops soil microbial biomass C and N decreased distinctly from seedling to grain-forming stages, and then increased to the maximum at crop maturity. Crop roots, however, showed reverse trend through the cropping period, suggesting strong competition between microbial biomass and crop roots for available nutrients. It is concluded that both resource quality and crop roots had distinct effect on soil microbial biomass and combined application of Sesbania shoot and wheat straw was most effective in sustained build up of microbial biomass through the annual cycle.