Comparison and improvement of methods for determining soil dehydrogenase activity by using triphenyltetrazolium chloride and iodonitrotetrazolium chloride

The triphenyltetrazolium chloride (TTC) method described by Thalmann (1968) and the iodonitrotetrazolium chloride (INT) method described by Spothelfer-Magaña and Thalmann (1992), used for measuring soil dehydrogenase activity, have been modified to overcome some methodical short-comings. Absorption maxima of 485 nm for triphenylformazan dissolved in acetone, 491 nm for iodonitrotetrazolium formazan (INTF) dissolved in tetrahydrofuran and 455 nm for INTF dissolved in N,N-dimethylformamide are recommended for measuring wavelengths. Extracting triphenylformazan twice with acetone is less toxic and proved to be at least as efficient as extraction with a mixture of 90% acetone and 10% carbon tetrachloride (Thalmann 1968 method). Tetrahydrofuran and dimethylformamide were equally good in extracting INTF from soils, but the former was less toxic. Anaerobic incubation resulted in the formation of higher amounts of triphenylformazan and INTF as well as reduced standard error. Both TTC and INT reduction showed high reproducibility and good differentiation of the microbial activity of six soils. For several reasons (more easily determined substrate dose depending on different soil types, better reduction, shorter incubation time), INT reduction seems to be a more suitable method of measuring soil microbial activity than TTC reduction.     

Soil microbial biomass and activities in a Japanese Andisol as affected by controlled release and application depth of urea

This experiment was conducted in maize field plots to study the effects of controlled release and application depth of urea on soil microbial biomass and activities at two depths of surface soil of a Japanese Andisol from June to September, 2001. Three N amendment treatments and a Control were included in this experiment: deep application (8 cm) of controlled release urea; deep application (8 cm) of conventional urea; surface application of conventional urea; Control, without N application. Prior to this experiment, the field plots received the same N fertilizer treatments for two consecutive years under maize/barley rotation. Soil microbial biomass, dehydrogenase and nitrification activities exhibited great vertical and temporal variations during the maize growth season, and the microbial biomass was significantly correlated to soil water-filled pore space (p<0.01). N fertilization did not significantly affect the microbial biomass, but greatly increased the dehydrogenase and nitrification activities. The increase in the microbial activities following N fertilization was not attributed to the increase in microbial biomass but to the increase in intrinsic microbial activities. Controlled release urea was found to continuously affect the dehydrogenase activity over a shorter distance, while conventional urea could greatly increase the enzyme activity for a shorter period of time. Both controlled release and deep application of urea had potentials to reduce the nitrification activity and suggested that the nitrate production might be decreased in 0–10 cm surface soil. Deep application of urea increased aboveground N uptake by maize and then the recovery rate of N fertilizer, whereas controlled release of urea greatly increased grain yield and N uptake by grain.     

The short-term effects of cessation of fertiliser applications,liming, and grazing on microbial biomass and activity in a reseeded upland grassland soil

A field study was conducted to determine the influence of a short-term (2 year) cessation of fertiliser applications, liming, and sheep-grazing on microbial biomass and activity in a reseeded upland grassland soil. The cessation of fertiliser applications (N and NPK) on a limed and grazed grassland had no effect on microbial biomass measurements, enzyme activities, or respiration. Withholding fertiliser and lime from a grazed grassland resulted in significant reductions in both microbial biomass C (P<0.05) and dehydrogenase activity (P<0.05) by approximately 18 and 21%, respectively. The removal of fertiliser applications, liming, and grazing resulted in even greater reductions in microbial biomass C (44%, P<0.001) and dehydrogenase activity (31%, P<0.001), and significant reductions in microbial biomass N (P<0.005), urease activity (P<0.05), phosphatase activity (P<0.001), and basal respiration (P<0.05). The abundance of culturable bacteria and fungi and the soil ATP content were unaffected by changes in grassland managements. With the cessation of liming soil pH fell from 5.4 to 4.7, and the removal of grazing resulted in a further reduction to pH 4.5. A significant negative linear relationship (r ²=0.97; P<0.01) was found between increasing soil acidity and dehydrogenase activity. Possible mechanisms influencing these changes are discussed.     

The use of biological methods to determine the microbiological activity of soils under cultivation

Summary In Ap horizons of typical arable soils under cereals in Northwest Germany, biological activity was estimated by measuring microbial activity. Twelve soils on local farms and six soils on a research farm were analysed. Microbial biomass, dehydrogenase activity, and alkaline phosphatase activity were compared with the biological availability of P, an index describing the relationship among several P fractions that has been used in ecological agriculture. The correlation between the microbial biomass and dehydrogenase and alkaline phosphatase activity was strong but the correlation between the biological availability of P and the enzyme activities was weak. In contrast, in the farm fields, there was a significant correlation between the microbial biomass and the biological availability of P. The correlation between the biological availability of P and pH was highly significant (r=0.65–0.93^***). Explanations for these correlations are discussed and proposals for further investigations are made. (1) Is the pH effect a direct chemical one or an indirect biological one? (2) Which soil organisms affect the biological availability of P in contrast to the microbial biomass, dehydrogenase activity, and alkaline phosphatase activity? (3) Is the method suitable for the investigation of all arable soils?     

Measurement of dehydrogenase activity using 2-p-iodophenyl-3-p-nitrophenyl-5-phenyltetrazolium chloride (INT) in the presence of copper

This paper reports on the adverse effect of Cu on the use of 2-p-iodophenyl-3-p-nitrophenyl-5-phenyltetrazolium chloride (INT) as a substrate for measuring microbial dehydrogenase activity (DHA). The presence of Cu affected the absorbance of the reaction product 2-p-iodophenyl-3-p-nitrophenyl-5-phenyltetrazolium formazan. The experiment provides evidence that the use of INT is an unreliable measure of DHA activity in soils contaminated with Cu.     

Soil microbial biomass and enzyme activity in subarctic agricultural and forest soils

Summary Microbial biomass, activities of dehydrogenase, phosphatase, and urease, and numbers of ammonium oxidizers were determined at monthly intervals on soil samples obtained from an on-going tillage residue-management study during the summers of 1985 and 1986. The site was cleared of black spruce (Picea mariana, Mill.) in 1979 and has been planted to spring barley (Hordeum vulgare) since 1982. Tillage treatments were no-tillage or disked twice, and residuemanagement treatments were removal of stubble and loose straw or leaving all straw on the plots. Microbial biomass and enzyme activities were moderate to high in the Ap horizon but very low in the B horizon. There was no difference in any parameter measured due to tillage or residue management. In 1986, comparisons were made between the Ap horizon and the agricultural soil and the A horizon of the soil beneath an adjacent black-spruce forest. Total microbial biomass and enzyme activities were generally greater in the forest soil than in the agricultural soil. However, specific activity of the biomass was generally greater in the agricultural soil. Soil microbial biomass and urease activities of both agricultural and forest soils were similar to those reported for warmer climates, but dehydrogenase activity was higher and phosphatase was lower.     

Grazing and plant-canopy effects on semiarid soil microbial biomass and respiration

The major objectives of this study were to determine the influence of grazing on the soil microbial biomass and activity in semiarid grassland and shrubland areas and to quantify the canopy effect (the differences in soil microbial biomass and activities between soils under plant canopies and soils in the open between plants). We also quantified changes in microbial biomass and activity during seasonal transition from dry to moist conditions. Chronosequences of sites withdrawn from grazing for 0, 11, and 16 years were sampled in a grassland (Bouteloua spp.) area and a shrubland (Atriplex canescens) area on and near the Sevilleta National Wildlife Reguge in central New Mexico, USA. Samples were obtained from beneath the canopies of plants (Yucca glauca in the grassland and A. canescens in the shrubland) and from open soils; they were collected three times during the spring and summer of a single growing season. Organic C, soil microbial biomass C, and basal respiration rates (collectively called the soil C triangle) were measured. We also calculated the microbial: organic C ratio and the metabolic quotient (ratio of respiration to microbial C) as measures of soil organic C stability and turnover. Although we had hypothesized that individual values of the soil C triangle would increase and that the ratios would decrease with time since grazing, differences in microbial parameters between sites located along the chronosequences were generally not significant. Grazing did not have a consistion effect on organic C, microbial C, and basal respiration in our chronosequences. The microbial: organic C ratio and the metabolic quotient generally increased with time since grazing on the shrubland chronosequence. The microbial: organic C ratio decreased with time since grazing and the metabolic quotient increased with time since grazing on the grassland chronosequence. The canopy effect was observed at all sites in nearly all parameters including organic C, microbial C, basal respiration, the microbial: organic C ratio, and the metabolic quotient which were predominantly higher in soils under the canopies of plants than in the open at all sites. Microbial biomass and activity did not increase during the experiment, even though the availability of moisture increased dramatically. The canopy effects were approximately equal on the shrubland and grassland sites. The microbial: organic C ratios and the metabolic quotients were generally higher in the shrubland soils than in the grassland soils.     

The use of microbial parameters in monitoring soil pollution by heavy metals

Microbial parameters appear very useful in monitoring soil pollution by heavy metals, but no single microbial parameter can be used universally. Microbial activities such as respiration, C and N mineralization, biological N₂ fixation, and some soil enzymes can be measured, as can the total soil microbial biomass. Combining microbial activity and population measurements (e.g., biomass specific respiration) appears to provide more sensitive indications of soil pollution by heavy metals than either activity or population measurements alone. Parameters that have some form of internal control, e.g., biomass as a percentage of soil organic matter, are also advantageous. By using such approaches it might be possible to determine whether the natural ecosystem is being altered by pollutants without recource to expensive and long-running field experiments. However, more data are needed before this will be possible. Finally, new applications of molecular biology to soil pollution studies (e.g., genetic fingerprinting) which may also have value in the future are considered.     

Activity and biomass of soil microorganisms at different depths

We measured microbial biomass C and soil organic C in soils from one grassland and two arable sites at depths of between 0 and 90 cm. The microbial biomass C content decreased from a maximum of 1147 (0–10 cm layer) to 24 g g^-1 soil (70–90 cm layer) at the grassland site, from 178 (acidic site) and 264 g g^-1 soil (neutral site) at 10–20 cm to values of between 13 and 12 g g^-1 soil (70–90 cm layer) at the two arable sites. No significant depth gradient was observed within the plough layer (0–30 cm depth) for biomass C and soil organic C contents. In general, the microbial biomass C to soil organic C ratio decreased with depth from a maximum of between 1.4 and 2.6% to a minimum of between 0.5 and 0.7% at 70–90 cm in the three soils. Over a 24-week incubation period at 25°C, we examined the survival of microbial biomass in our three soils at depths of between 0 and 90 cm without external substrate. At the end of the incubation experiment, the contents of microbial biomass C at 0–30 cm were significantly lower than the initial values. At depths of between 30 and 90 cm, the microbial biomass C content showed no significant decline in any of the four soils and remained constant up to the end of the experiment. On average, 5.8% of soil organic C was mineralized at 0–30 cm in the three soils and 4.8% at 30–90 cm. Generally, the metabolic quotient qCO₂ values increased with depth and were especially large at 70–90 cm in depth.     

Ergosterol and microbial biomass relationship in soil

Ergosterol and microbial biomass C were measured in 26 arable, 16 grassland and 30 forest soils. The ergosterol content ranged from 0.75 to 12.94 g g^-1 soil. The geometric mean ergosterol content of grassland and forest soils was around 5.5 g g^-1, that of the arable soils 2.14 g g^-1. The ergosterol was significantly correlated with biomass C in the entire group of soils, but not in the subgroups of grassland and forest soils. The geometric mean of the ergosterol: microbial biomass C ratio was 6.0 mg g^-1, increasing in the order grassland (5.1), arable land (5.4) and woodland (7.2). The ergosterol:microbial biomass C ratio had a strong negative relationship with the decreasing cation exchange capacity and soil pH, indicating that the fungal part of the total microbial biomass in soils increased when the buffer capacity decreased. The average ergosterol concentration calculated from literature data was 5.1 mg g^-1 fungal dry weight. Assuming that fungi contain 46% C, the conversion factor from micrograms ergosterol to micrograms fungal biomass C is 90. For soil samples, neither saponification of the extract nor the more effective direct saponification during extraction seems to be really necessary.     

Substrate type,temperature, and moisture content affect gross and net N mineralization and nitrification rates in agroforestry systems

Accurate prediction of soil N availability requires a sound understanding of the effects of environmental conditions and management practices on the microbial activities involved in N mineralization. We determined the effects of soil temperature and moisture content and substrate type and quality (resulting from long-term pasture management) on soluble organic C content, microbial biomass C and N contents, and the gross and net rates of soil N mineralization and nitrification. Soil samples were collected at 0–10 cm from two radiata pine (Pinus radiata D. Don) silvopastoral treatments (with an understorey pasture of lucerne, Medicago sativa L., or ryegrass, Lolium perenne L.) and bare ground (control) in an agroforestry field experiment and were incubated under three moisture contents (100, 75, 50% field capacity) and three temperatures (5, 25, 40 °C) in the laboratory. The amount of soluble organic C released at 40 °C was 2.6- and 2.7-fold higher than the amounts released at 25 °C and 5 °C, respectively, indicating an enhanced substrate decomposition rate at elevated temperature. Microbial biomass C:N ratios varied from 4.6 to 13.0 and generally increased with decreasing water content. Gross N mineralization rates were significantly higher at 40 °C (12.9 g) than at 25 °C (3.9 g) and 5 °C (1.5 g g^–1 soil day^–1); and net N mineralization rates were also higher at 40 °C than at 25 °C and 5 °C. The former was 7.5-, 34-, and 29-fold higher than the latter at the corresponding temperature treatments. Gross nitrification rates among the temperature treatments were in the order 25 °C >40 °C >5 °C, whilst net nitrification rates were little affected by temperature. Temperature and substrate type appeared to be the most critical factors affecting the gross rates of N mineralization and nitrification, soluble organic C, and microbial biomass C and N contents. Soils from the lucerne and ryegrass plots mostly had significantly higher gross and net mineralization and nitrification rates, soluble organic C, and microbial biomass C and N contents than those from the bare ground, because of the higher soil C and N status in the pasture soils. Strong positive correlations were obtained between gross and net rates of N mineralization, between soluble organic C content and the net and gross N mineralization rates, and between microbial biomass N and C contents.     

Effects of past and current crop management on soil microbial biomass and activity

Because soil biota is influenced by a number of factors, including land use and management techniques, changing management practices could have significant effects on the soil microbial properties and processes. An experiment was conducted to investigate differences in soil microbiological properties caused by long- and short-term management practices. Intact monolith lysimeters (0.2 m² surface area) were taken from two sites of the same soil type that had been under long-term organic or conventional crop management and were then subjected to the same 2.5-year crop rotation [winter barley (Hordeum vulgare L.), maize (Zea mais L.), lupin (Lupinus angustifolius L.), and rape (Brassica napus L. ssp. oleifera)] and two fertilizer regimes (following common organic and conventional practices). Soil samples were taken after crop harvest and analyzed for microbial biomass C and N, microbial activity (fluorescein diacetate hydrolysis, arginine deaminase activity, and dehydrogenase activity), and total C and N. The incorporation of the green manure stimulated growth and activity of the microbial communities in soils of both management histories. Soil microbial properties did not show any differences between organically and conventionally fertilized soils, indicating that crop rotation and plant type had a larger influence on the microbial biomass and enzyme activities than fertilization. Initial differences in microbial biomass declined, while the effects of farm management history were still evident in enzyme activities and total C and N. Links between enzyme activities and microbial biomass C varied depending on treatment, indicating differences in microbial community composition.     

Continuous defoliation of perennial ryegrass (Lolium perenne) and white clover (Trifolium repens) and associated changes in the composition and activity of the microbial population of an upland grassland soil

A microcosm study was conducted to investigate the effect of continuons plant defoliation on the composition and activity of microbial populations in the rhizosphere of perennial ryegrass (Lolium perenne) and white clover (Trifolium repens). Continuons defoliation of ryegrass and clover resulted in sigmficant (P <0.01) increases in soil microbial biomass, although whilst increases were measured from day 2 in soil sown with clover significant increases were only seen from day 21 in soil sown with ryegrass. These increases were paralleled, from day 10 onwards, by increases in the numbers of culturable bacteria. Numbers ofPsendomonas spp. also increased in the later stages of the study. No influence on culturable fungal populations was detected. Whilst shifts in the composition of the microbial populations were measured in response to defoliation there was little effect on microbial activity. No changes in either dehydrogenase activity or microbial respiration in the rhizosphere of ryegrass or clover were measured in response to defoliation, but both dehydrogenase activity and microbial respiration were greater in ryegrass than clover when values over the whole study were combined. Continuous defoliation resulted in significant (P <0.001) reductions in the root dry weight of ryegrass and clover, of the order 19% and 16%, respectively.     

管理措施对水稻土中电子传输体系活性的影响

Electron transport system(ETS)/dehydrogenase activity in a paddy field soil was measured under a variety of incubation conditions using the reduction of 2-(p-iodophenyl-3-(p-nitrophenyl)-5-phenyl tetrazolium chloride(INT) to iodonitrotetrazolium formazan(INTF).The results exhibited a high positive correlation between the ETS activity and the incubation temperature and soil moisture,Dehydrogenase/ETS activity displayed a negative correlation with insecticide concentrations,and the activity affected adversely as the concentration of the insecticide increased.The hiher doses,5 and 10 field rates(1 field rate=1500mL ha^-1),of insecticide significantly inhibited ETS activity,while lower rates failed to produce any significant reducing effect,Inorganic N( as urea) of concentrations from 0 to 100ug N g^-1 soil showed a positive response to ETS activity,However,at concentrations of 200 and 400ug N g^-1,the activity was reduced significantly.     

Soil enzymatic response to addition of municipal solid-waste compost

Modifications of soil microbiological activity by the addition of municipal solid-waste compost were studied in laboratory incubations. Three composts were compared, one lumbricompost and two classical composts with different maturation times. Organic C mineralization and nine enzyme activities (dehydrogenase, peroxidase, cellulase, -glucosidase, -galactosidase, N-acetyl--glucosaminidase, protease, amidase, and urease) were determined in the composts and the amended soil. Initial enzyme activities varied in the soil according to the sampling date (winter or summer) and were greater in the composts than in the soil, except for urease. Generally, the youngest compost exhibited greater activity than the oldest one. In the amended soil, the composts did not increase enzyme activity in an additive way. Dehydrogenase, the only strictly endocellular enzyme, was the only one for which the activity in the amended soil increased significantly in proportion to the addition of compost. During the incubations, C mineralization and dehydrogenase activity were significantly correlated, indicating that dehydrogenase was a reliable indicator of global microbial activity. Peroxidase activity in the soil remained constant, but increased in the composts and amended soil. Addition of the oldest compost had no effect on the activity of the C cycle enzymes, but the youngest compost increased creased soil activity at the higher application rate. Enzymes of the N cycle were stimulated by all compost amendments, but the increase was only transient for amidase and urease. Lumbricomposting had no marked effect on compost enzyme activity, either before or during the incubation.     

Microbial response of an acid forest soil to experimental soil warming

 Effects of increased soil temperature on soil microbial biomass and dehydrogenase activity were examined on organic (O) horizon material in a low-elevation spruce-fir ecosystem. Soil temperature was maintained at 5  °C above ambient during the growing season in the experimental plots, and soil temperature, moisture, microbial biomass, and dehydrogenase activity were measured during the experiment. An incubation study was also conducted under three temperature regimes, 5, 15, and 25  °C, and under four moisture regimes of 20, 120, 220, and 320% to further evaluate these environmental factors on dehydrogenase activity and microbial biomass. Soil moisture content and microbial biomass controls were significantly lower (30% and 2 μg g^–1 soil, respectively) in the heated plots during the treatment period, suggesting that moisture content was important in controlling microbial biomass. In the incubation study, temperature appeared more important than moisture in controlling microbial biomass and dehydrogenase activity. Increasing temperature between 5  °C and 25  °C resulted in significant decreases in microbial biomass and dehydrogenase activity. Received: 7 August 1998     

Soil microbial biomass, dehydrogenase activity, bacterial community structure in response to long-term fertilizer management

This study describes the effects of balanced versus nutrient-deficiency fertilization on soil microbial biomass, activity, and bacterial community structure in a long-term (16 years) field experiment. Long-term fertilization greatly increased soil microbial biomass C and dehydrogenase activity, except that the P-deficiency fertilization had no significant effect. Organic manure had a significantly greater (P<0.05) impact on the biomass C and the activity, compared with mineral fertilizers. Microbial metabolic activity (dehydrogenase activity per microbial biomass C) was significantly higher (P<0.05) under balanced fertilization than under nutrient-deficiency fertilization. General bacterial community structure was analyzed by PCR-denaturing gradient gel electrophoresis (DGGE) targeting eubacterial 16S rRNA gene. Mineral fertilization did not affect the DGGE banding pattern, while specific DGGE band was observed in organic manure-fertilized soils. Phylogenetic analysis showed that the change of bacterial community in organic manure-fertilized soil might not be because of the direct influence of the bacteria in the compost, but because of the promoting effect of the compost on the growth of an indigenous Bacillus sp. in the soil. We emphasize the importance of balanced-fertilization, as well as the role of P, in maintaining soil organic matter, and promoting the biomass and activity of microorganisms.     

Microbial activities during the early stage of laboratory decomposition of tropical leaf litters: the effect of interactions between litter quality and exogenous inorganic nitrogen

The comparative decomposition of tropical leaf litters (e.g. Andropogon gayanus, Casuarina equisetifolia, Faidherbia albida) of different qualities was investigated under laboratory conditions during a 60-day incubation period conducted with a typical oxisol. Total CO₂-C, soil inorganic N, microbial biomass (fumigation-extraction), -glucosidase and dehydrogenase activities were determined over the incubation to assess how they responded to the addition of inorganic N (+N). Cumulative CO₂-C evolved from the litter-amended soils was higher than that recorded for the unamended control soil. For the unfertilized treatment (0 N), correlation coefficients calculated between initial chemical data and CO₂ flux during the first day of incubation were r =0.963 for water soluble-C and 0.869 for soluble carbohydrates (P <0.05). At the end of the incubation, the amounts of CO₂-C in the F. albida- and A. gayanus-amended soils were higher than that in the C. equisetifolia-amended treatment. Cumulative net N immobilization increased during the first 30 days of incubation, the amounts being similar for A. gayanus- and C. equisetifolia-amended soil and higher than that recorded in the F. albida-amended treatment. Soil microbial biomass and enzyme activities increased in the litter-amended soils during the first 15 days of incubation and decreased (except for the dehydrogenase activity) thereafter. The addition of inorganic N modified the patterns of CO₂-C respiration and net N immobilization. The magnitude of these modifications varied according to the litter quality. The use of an accurate indicator based on several litter components to predict the amplitude of organic material decomposition is discussed.     

The impacts of individual plant species on rhizosphere microbial communities in soils of different fertility

To investigate the effects of individual plant species on microbial community properties in soils of differing fertility, a microcosm experiment was carried out using plant species representative of the dominant flora in semi-fertile temperate grasslands of northern England. Soil microbial biomass and activity were found to be significantly greater in the more fertile, agriculturally improved soil than in the less productive unimproved meadow soil. Differences in microbial community structure were also evident between the two soils, with fungal abundance being greater in the unimproved soil type. Individual plant species effects significantly differed between the two soils. Holcus lanatus and Anthoxanthum odoratum stimulated microbial biomass in the improved soil type, but negatively affected this measure in the unimproved soil. In both soil types, herb species generally had negative effects on microbial biomass. Patterns for microbial activity were less consistent, but as with microbial biomass, A. odoratum and H. lanatus promoted respiration, whereas the herbs negatively affected this measure. All plant species grown in the improved soil increased the abundance of fatty acids synthesised by bacteria (bacterial phospholipid fatty acid analysis) relative to bare soil, but they negatively impacted on this group of fatty acids in unimproved soil. Similarly, the abundance of the fungal fatty acid 18:26 was increased by all plants in the more fertile improved soil only, albeit non-significantly. Our data indicate that effects of plant species on microbial properties differ markedly in soils of differing fertility, making general predictions about how individual plants impact on soil properties difficult to make.     

Organic matter, microbial biomass and enzyme activity of soils under different crop rotations in the tropics

Soil organic matter level, soil microbial biomass C, ninhydrin-N, C mineralization, and dehydrogenase and alkaline phosphatase activity were studied in soils under different crop rotations for 6 years. Inclusion of a green manure crop of Sesbania aculeata in the rotation improved soil organic matter status and led to an increase in soil microbial biomass, soil enzyme activity and soil respiratory activity. Microbial biomass C increased from 192 mg kg^–1 soil in a pearl millet-wheat-fallow rotation to 256 mg kg^–1 soil in a pearl millet-wheat-green manure rotation. Inclusion of an oilseed crop such as sunflower or mustard led to a decrease in soil microbial biomass, C mineralization and soil enzyme activity. There was a good correlation between microbial biomass C, ninhydrin-N and dehydrogenase activity. The alkaline phosphatase activity of the soil under different crop rotations was little affected. The results indicate the green manuring improved the organic matter status of the soil and soil microbial activity vital for the nutrient turnover and long-term productivity of the soil. Received: 7 January 1996