Comparison of extraction methods for recovery of extracellular β-glucosidase in two different forest soils

A study was made of the efficiency of three different extractants, 0.1 M sodium pyrophosphate (pH 7), 67 mM phosphate buffer (pH 6) and 0.5 M potassium sulphate (pH 6.6), in recovering the protein quantity and the β-glucosidase enzyme activity from two natural forest soils: (1) an Inceptisoil located in Tuscany (Italy) in a mild Mediterranean climate, and (2) a Lithic Calcixeroll soil located in Murcia (south-east of Spain) in a dry-semiarid climate. The pyrophosphate was used to determine the activity of extracellular-humic-bound proteins, while the phosphate buffer and potassium sulphate were used to extract dissolved extracellular proteins. The latter extractant, after chloroform fumigation, was also used to measure total proteins in soil. A preliminary screening, using SDS-PAGE in one dimension, was also carried out in order to optimize the separation condition of soil proteins extracted with different buffers. To remove the interfering co-extracted substances (humic acid) a purification step using a column packed with insoluble polyvinylpyrrolidone was performed. The highest β-glucosidase activity was recovered in the pyrophosphate extract, thus confirming its capability of extracting humic-bound β-glucosidase enzyme in a stable and active form. The extractants performed differently with the two soil types and band patterns obtained with SDS-PAGE were extractant-specific, demonstrating that each was selective for a particular class of proteins. Surprisingly, protein bands were also obtained using pyrophosphate, in spite of the very dark extract colour due to the presence of humic substances.     

Extraction and properties of phosphodiesterase from a forest soil

Phosphodiesterase together with brown-coloured compounds was extracted from a forest soil using 0.1 M phosphate buffer (pH 7). Use of KCl and EDTA with the buffer facilitated phosphodiesterase extraction. Distilled water extracted little enzyme activity. A curvilinear relationship such as the Langmuir type was found between solution volume and phosphodiesterase activity of the extract. The results implied that the phosphodiesterase extracted was extracellular and was adsorbed on the surface of soil particles by ionic bonding. Brown-coloured compounds in the extract were removed by precipitation with protamine sulfate. The phosphodiesterase activity of the extract treated with protamine sulfate was lost on keeping at 80°C for 10 min and was optimal at pH 5.2–6.0. The extract hydrolyzed either the 3'- or the 5'-phosphodiester bond of deoxythymidine p-nitrophenyl phosphate.     

Pre-lysis washing improves DNA extraction from a forest soil

A pre-lysis buffer washing procedure was introduced to DNA extraction from a forest soil with high organic matter and iron oxide contents. Sodium phosphate of 0.1 M (pH 7.5) was used as a buffer to wash soil samples when subsequent lysis buffer was phosphate, and 20 mM EDTA (pH 7.5) was used when subsequent lysis buffer included EDTA. Initial experiments were not successful because the DNA extracts could not be amplified by polymerase chain reaction (PCR). The consideration of introducing a pre-lysis washing procedure was based on the idea that the washing should promote soil dispersion and homogeneity, decrease DNA adsorption by soil components (e.g. iron oxides), and remove covalent cations and those easily-dissolving organic compounds from the soil samples. Results revealed that humic substance content decreased by 31%, but DNA yield increased by 24% in the DNA extracts of the pre-lysis washing procedures, compared to the non-washing procedures. DNA extracted by the pre-washing procedure needed less purification for subsequent 18S and 16S rDNA PCR amplifications. It was recommended that the pre-lysis buffer washing should be used for DNA extraction from those difficult environmental samples, such as the forest soil with high contents of organic matter and iron oxides.     

Extraction of β-glucosidase activity from PEA field soil

Brown compounds with β-glucosidase activity were extracted from a cultivated soil in 0.1 m phosphate buffer (pH 7), 0.3 m KCl and 10 mm EDTA. A curvilinear relationship of the Langmuir type was observed between the solution volume and β-glucosidase activity of the extract. The results indicated the β-glucosidase was extracellular and was adsorbed on the surfaces of soil particles by ionic bonds. The preparation was treated with protamine sulfate and its enzymatic properties were investigated. The activity of the preparation was optimal at pH 5.4 and was lost after 10 min at 80°C. The extract hydrolyzed p-nitrophenyl β-glucoside(100), phenyl β-glucoside(19), salicin(8), amygdalin(33), cellobiose(52) and gentiobiose(55) (relative activities shown in parentheses). But, the extract had no effect on methyl β-glucoside and phlorizin. The substrate specificity and optimum pH of the enzymatic activity of the soil extract was similar to those of β-glucosidases from various fungi.     

Available organic nitrogen in temperate, subtropical, and tropical soils extracted with different solutions

Extraction of organic N by chemical solutions has been used to assess the amount of available N in soil. We tested the efficiency of several solutions in extracting organic N from tropical, subtropical and temperate soils. A conventional 0.067 M phosphate buffer successfully extracted organic N from all 23 soils examined. High-performance size exclusion chromatograms showed a single peak at about 7,800 Da for all phosphate buffer extracts irrespective of soil types. The peak area correlated with the organic N concentration of extracts. Tropical soils had lower retention of organic N than other soils according to the conventional and sequential extraction with phosphate buffer. Organic N extracted with sulfuric acid was significantly (P < 0.001) correlated with the amount of extracted Fe, suggesting that Fe might play a role in the retention of organic N in soil.     

Phosphorus availability to corn from wood ash-amended soils

Wood ash is a residual material produced during biomass burning. In the northeastern United States up to 80 % of the ash is spread on agricultural lands as a liming amendment with the remainder being disposed of in landfills. As well as raising soil pH, wood ash also adds plant nutrients to soil. This study is an examination of the plant availability of the P in 8 different soils amended with one wood ash. Plant availability was assessed by measuring the biomass and P concentration of corn (Zea mays) L.) plants grown in the greenhouse for 28 d in soil amended with either CaCO₃ (control), wood ash to supply 200 mg kg^?1 total P, or monocalcium phosphate (MCP) to supply 200 mg kg^?1 total P and CaCO₃. Both corn growth and P uptake were highest in the MCP treatments, intermediate in the wood ash treatments, and lowest in the controls for all soil types. The soil property which seemed to have the greatest influence on P availability was pH buffer capacity. The soils with the greatest capacity to buffer OH additions also tended to exhibit the greatest absolute P uptake from wood ash-amended soils and the greatest P uptake relative to that from MCP-amended soils. The ability of soil test extractants to predict uptake of P in the three soil treatments was examined. A buffered ammonium acetate extradant overestimated P availability in the ash-amended soils relative to the MCP-amended soils. An unbuffered, acid, fluoride-containing extract provided a measure of P levels that was consistent with P uptake from all soil treatments. In this study the predictive relationship was as follows: P uptake = 0.017× (Bray P, mg kg^?1) + 1.19; r = 0.81.     

土壤组分对广东省酸性水稻土磷吸附参数的影响

Soil components affecting phosphate sorption parameters were studied using acid paddy soils derived from basalt, granite, sand-shale and the Pearl River Delta sediments, respectively, in Guangdong Province.For each soil, seven 2.50 g subsamples were equilibrated with 50 mL 0.02 mol L-1 (pH=7.0) of KCl containing 0, 5, 10, 15, 25, 50 and 100 ng P kg-1, respectively, in order to derive P sorption parameters (P sorption maximum, P sorption intensity factor and maximum buffer capacity) by Langmuir isotherm equation. It was shown that the main soil components influencing phosphate sorption maximum (Xm) included soil clay, pH,amorphous iron oxide (Feo) and amorphous aluminum oxide (Alo), with their effects in the order of Alo ＞Feo ＞ pH ＞ clay. Among these components, pH had a negative effect, and the others had a positive effect.Organic matter (OM) was the only soil component influencing P sorption intensity factor (K). The main components influencing maximum phosphate buffer capacity (MBC) consisted of soil clay, OM, pH, Feo and Alo, with their effects in the order of Alo ＞ OM ＞ pH ＞ Feo ＞ clay. Path analysis indicated that among the components with positive effects on maximum phosphate buffer capacity (MBC), the effect was in the order of Alo ＞ Feo ＞ Clay, while among the components with negative effects, OM ＞ pH. OM played an important role in mobilizing phosphate in acid paddy soils mainly through decreasing the sorption intensity of phosphate by soil particles.     

Soil properties related to phosphate buffering in calcareous soils

Abstract

In a group of 24 related calcareous soils, derived from Jurassic oolitic limestone, there was marked variability (13‐fold) in phosphate buffering when expressed as the maximum buffer capacity. This variability was most closely related to the iron content and pH of the soils, and these together accounted for 72% of the variance. This percentage was not increased by including CaC0₃ content or organic matter, which were also correlated with the maximum buffer capacity. A high correlation with specific surface area of CaCO₃ was probably an indirect effect due to the high correlation between this variable and the Fe and pH of the soils.

The equilibrium buffer capacity, which is the traditional measure of phosphate buffering, was less variable but quite unrelated to all the soil properties measured except the soil surface area. However the maximum buffer capacity and quantity of adsorbed P together accounted for 63% of the variance in this parameter.     

Hydrolysis of cellobiose by β-glucosidase in the presence of soil minerals - Interactions at solid-liquid interfaces and effects on enzyme activity levels

Extracellular enzymatic activities in soils are essential for the cycling of organic matter. These activities take place in multiphase environments where solid phases profoundly affect biocatalytic activities. Aspergillus niger is ubiquitous in soils; its β-glucosidase plays an important role in the degradation of cellulose, and therefore in the global carbon cycle and in the turnover of soil organic matter. However, the information on the interactions of this protein with soil minerals is very limited, and even less is known about their consequences for the hydrolysis of the natural substrate cellobiose. We therefore characterised the sorptive interactions of this enzyme with the soil minerals montmorillonite, kaolinite and goethite and quantified the resulting changes in the hydrolysis rate of cellobiose. Fractions of adsorbed protein, and the resulting catalytic activity loss, were lower for montmorillonite than for kaolinite and goethite at given experimental conditions; adsorption was 9.7 ± 7.3% for montmorillonite, 70.3 ± 3.1% for kaolinite and 71.4 ± 1.8% for goethite, respectively. Adsorption of the protein to the minerals caused a total decrease in the catalytic activity of 18.8 ± 3.4% for kaolinite and 17.9 ± 4.7% for goethite whereas it was not significant for montmorillonite. The average catalytic activity lost by the pool of adsorbed molecules was 26.8% for kaolinite and 25.0% for goethite. Both the amount of adsorbed protein and the resulting loss of catalytic activity were found to be independent of the specific surface areas yet were influenced by the electrical properties of the mineral surfaces. Under the experimental conditions, montmorillonite and kaolinite are negatively charged whereas goethite is positively charged. However, because of the adsorption of phosphate anions from the buffer, a charge reversal took place at the surface of goethite. This was confirmed by zeta (ζ)-potential measurements in phosphate buffer, revealing negative values for all the tested minerals. Indeed goethite interacted with the enzyme as a negatively charged surface: the amount of adsorbed protein and the resulting catalytic activity loss were very similar to those of kaolinite. Our results show that, even if an important fraction of β-glucosidase is adsorbed to the minerals, the catalytic activity is largely retained. We suggest that this strong activity retention in presence of soil minerals results from a selective pressure on A. niger, which benefits from the activity of the adsorbed, and thus stabilized, enzyme pool.     

Phosphorus fractionation in lowland tropical rainforest soils in central Panama

Phosphorus availability is commonly assumed to limit productivity in lowland tropical rainforests, yet there is relatively little information on the chemical forms of soil phosphorus in such ecosystems. We used the Hedley sequential fractionation scheme to assess phosphorus chemistry in five soils supporting tropical rainforest on Barro Colorado Island, Republic of Panama. The soils represented a range of orders (Inceptisols, Alfisols, and Oxisols) formed on contrasting geological substrates and topography, but under uniform climate and vegetation. Total phosphorus in surface horizons ranged between 315 and 1114 mg P kg^− 1, being lowest on a soil derived from marine sediments and highest on soils derived from andesite. The majority of the phosphorus occurred in recalcitrant forms, although between 14% and 39% occurred as organic phosphorus. Readily-available phosphate, as extracted by anion-exchange membranes, occurred in small concentrations (4–13 mg P kg^− 1), although labile phosphorus, defined as phosphate extracted by anion-exchange membrane plus inorganic and organic phosphorus extracted by 0.5 M NaHCO₃, accounted for between 4.7% and 11.4% of the total soil phosphorus. Our results indicate a strong control of geology and topography on soil phosphorus in tropical rainforests, which may have important implications for understanding the diversity and distribution of plant species in such ecosystems. Further, some of the most common soils on Barro Colorado Island, including those on the 50 ha forest dynamics plot, are rich in phosphorus despite their relatively advanced stage of pedogenesis.     

Quantification and bioavailability of scyllo-inositol hexakisphosphate in pasture soils

The recent identification of scyllo-inositol hexakisphosphate in alkaline soil extracts by solution ³¹P NMR spectroscopy allowed us to investigate this compound in soils by re-analyzing spectra from two previously published studies. Concentrations of scyllo-inositol hexakisphosphate in 29 temperate pasture soils from England and Wales ranged between 11 and 130 mg P kg⁻¹ soil and accounted for between 4 and 15% of the soil organic phosphorus. The ratio of scyllo-inositol hexakisphosphate to myo-inositol hexakisphosphate ranged between 0.29 and 0.79. In a 10 month pot experiment with six grassland soils from New Zealand, growth of pine seedlings (Pinus radiata D. Don) decreased scyllo-inositol hexakisphosphate concentrations by between 10 and 46%. Growth of ryegrass (Lolium perenne L.) decreased scyllo-inositol hexakisphosphate in three low-nutrient soils by 5-21%, but increased it in three other soils by 11-16%. We conclude that scyllo-inositol hexakisphosphate is an important component of soil organic phosphorus with potential ecological significance.     

Isolating the influence of pH on the amounts and forms of soil organic phosphorus

Soil pH influences the chemistry, dynamics and biological availability of phosphorus (P), but few studies have isolated the effect of pH from other soil properties. We studied phosphorus chemistry in soils along the Hoosfield acid strip (Rothamsted, UK), where a pH gradient from 3.7 to 7.8 occurs in a single soil with little variation in total phosphorus (mean ± standard deviation 399 ± 27 mg P kg^?1). Soil organic phosphorus represented a consistent proportion of the total soil phosphorus (36 ± 2%) irrespective of soil pH. However, organic phosphorus concentrations increased by about 20% in the most acidic soils (pH < 4.0), through an accumulation of inositol hexakisphosphate, DNA and phosphonates. The increase in organic phosphorus in the most acidic soils was not related to organic carbon, because organic carbon concentrations declined at pH < 4.0. Thus, the organic carbon to organic phosphorus ratio declined from about 70 in neutral soils to about 50 in strongly acidic soils. In contrast to organic phosphorus, inorganic phosphorus was affected strongly by soil pH, because readily‐exchangeable phosphate extracted with anion‐exchange membranes and a more stable inorganic phosphorus pool extracted in NaOH–EDTA both increased markedly as soil pH declined. Inorganic orthophosphate concentrations were correlated negatively with amorphous manganese and positively with amorphous aluminium oxides, suggesting that soil pH influences orthophosphate stabilization via metal oxides. We conclude that pH has a relatively minor influence on the amount of organic phosphorus in soil, although some forms of organic phosphorus accumulate preferentially under strongly acidic conditions.     

Laccases in soil and the feasibility of their extraction

Because investigations had established the difficulty of isolating a laccase from native soil, experiments were made to overcome this problem. An extracellular laccase which had been isolated from the growth medium of the fungus Trametes versicolor was introduced into the soil. After a short incubation, approximately 80% of this activity could be recovered with water, aconitate or phosphate buffers as extradants. The enzyme could also be extracted from soils which had been inoculated with various laccase-producing fungi; maximum laccase formation was usually observed after about 30 days. Attempts were made to extract laccase activity from native soil with 10 different buffers, but only those buffers containing citrate ions yielded a positive result. It is unlikely that this activity is due to an enzyme, since it has been demonstrated that citrate ions and manganese form complexes which exhibit laccase-like activity. However, when soil was mixed with sand and placed inside a column, it was possible to elute laccase-like activity with several non-citrate buffers containing laccase substrates. This activity was found only in the eluents of fresh soils, but not in those of air-dried or autoclaved soils.     

基于不同方法测定土壤酸性磷酸酶活性的比较

土壤酸性磷酸酶与有机磷的矿化及植物的磷素营养关系最为密切。目前国内学者在测定酸性磷酸酶活性时主要参照关松荫《土壤酶及其研究法》中以磷酸苯二钠为基质的测定方法,而国外学者主要参照Dick《Methods of Soil Enzymology》中以对硝基苯磷酸二钠为基质的测定方法(PNPP)。但是,在以磷酸苯二钠为基质测定生成物的过程中,常出现显色程度不明显的问题;另外,采用不同基质测定酸性磷酸酶活性也造成了测定方法选择的困难。为合理选择土壤酸性磷酸酶活性的测定方法,本研究选用酸性、中性和碱性土壤各10个土样,分别采用以磷酸苯二钠为基质,且在显色阶段分别加入pH5.0醋酸盐缓冲液(DPP 1)和pH9.4硼酸盐缓冲液(DPP 2)的方法,以及PNPP方法测定土壤酸性磷酸酶活性。同时也研究了不同pH缓冲液和苯酚浓度对生成物显色反应的影响。结果表明:以磷酸苯二钠为基质、在显色反应阶段加入pH≤6的缓冲液时,苯酚和2,6-二溴苯醌氯亚胺不显色;当加入pH≥8的缓冲液时,两者之间显色且苯酚浓度和吸光值的Pearson相关系数极显著。这说明pH低是导致高苯酚浓度和2,6-二溴苯醌氯亚胺显色效果差的一个主要原因。此外,采用PNPP方法测定时,在酸性、中性和碱性土壤中,10个样本酸性磷酸酶活性的变异系数分别较DPP 2增加了70.04%、42.44%和21.17%;极差分别是DPP 2的27.18倍、26.85倍和39.43倍。总之,如果选用磷酸苯二钠为基质测定土壤酸性磷酸酶活性,应在显色阶段加入碱性硼酸盐缓冲液;选用对硝基苯磷酸二钠为基质,是更为简单和灵敏的方法。     

Effects of physicochemical interactions and microbial activity on the persistence of Cry1Aa Bt (Bacillus thuringiensis) toxin in soil

Genetically modified crops, that produce Cry insecticidal crystal proteins (Cry) from Bacillus thuringiensis (Bt), release these toxins into soils through root exudates and upon decomposition of residues. The fate of these toxins in soil has not yet been clearly elucidated. Persistence can be influenced by biotic (degradation by microorganisms) and abiotic factors (physicochemical interactions with soil components, especially adsorption). The aim of this study was to follow the fate of Cry1Aa Bt toxin in contrasting soils subjected to different treatments to enhance or inhibit microbial activity, in order to establish the importance of biotic and abiotic processes for the fate of Bt toxin. The toxin was efficiently extracted from each soil using an alkaline buffer containing a protein, bovine serum albumin, and a nonionic surfactant, Tween 20. The marked decline of extractable toxin after incubation of weeks to months was soil-dependent. The decrease of extractable toxin with incubation time was not related to microbial degradation but mainly to physicochemical interactions with the surfaces that may decrease immunochemical detectability or enhance protein fixation. Hydrophobic interactions may play an important role in determining the interaction of the toxin with surfaces.     

Increase in soil pH due to Ca-rich organic matter application causes suppression of the clubroot disease of crucifers

Clubroot disease of cruciferous plants caused by the soil-borne pathogen Plasmodiophora brassicae is difficult to control because the pathogen survives for a long time in soil as resting spores. Disease-suppressive and conducive soils were found during the long-term experiment on the impact of organic matter application to arable fields and have been studied to clarify the biotic and abiotic factors involved in the disease suppression. The fact that a large amount of organic matter, 400 t ha⁻¹ yr⁻¹ farmyard manure (FYM) or 100 t ha⁻¹ yr⁻¹ food factory sludge compost (FSC), had been incorporated for more than 15 yr in the suppressive soils and these soils showed higher pH and Ca concentration than the disease conducive soil led us to hypothesize that an increase in soil pH due to the long-term incorporation of Ca-rich organic matter might be the primary cause of the disease suppression. We have designed a highly reproducible bioassay system to examine this hypothesis. The suppressive and conducive soils were mixed with the resting spores of P. brassicae at a rate of 10⁶ spore g⁻¹ soil, and Brassica campestris was grown in a growth chamber for 8 d. The number of root hair infections was assessed on a microscope. It was found that the incorporation of FYM and FSC at 2.5% (w/w) to the conducive soil suppressed the infection and that the finer particles (?5 mm) of FSC inhibited the infection and increased soil pH more effectively. Neutralization of the conducive soil by Ca(OH)₂, CaCO₃ and KOH suppressed the infection, but the effectiveness of KOH was less than those of Ca(OH)₂ and CaCO₃. Acidification of the suppressive soils by H₂SO₄, promoted the infection. The involvement of soil biota in the disease suppression was investigated using the sterilized (γ-ray irradiation) suppressive soils with respect to soil pH. The γ-ray irradiation promoted the infection at pH 5.5, but no infection was observed at pH 7.4 irrespective of the sterilization status. All these observations suggest that soil pH is a major factor in disease suppression by organic matter application and that Ca and soil biota play certain roles in the suppression under the influence of soil pH.     

铁蟹过敏原的分离、鉴定和快速纯化

应用免疫印迹(Western blot) 的方法鉴定铁蟹过敏原组分,然后利用电泳洗脱的方法快速纯化主要过敏原。取磷酸盐缓冲液制备的铁蟹肉浸出液,经十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分离,测定各组分的相对分子量,然后利用18 例蟹过敏患者的血清进行免疫印迹,鉴定出主要和次要过敏原,利用普通垂直电泳槽快速洗脱主要过敏原蛋白,并鉴定活性。结果显示,SDS-PAGE显示铁蟹肉可辨条带有16条,相对分子质量在16.5～168 kD 之间。Western blotting结果表明,铁蟹肉浸出液共有10条过敏条带,其中相对分子质量为76kD的蛋白条带,阳性反应率为100%,纯化后获取了76 kD的主要过敏原,经过免疫印迹鉴定其具有免疫活性。表明相对分子质量76 kD的组分为铁蟹主要过敏组分,快速电洗脱可以纯化相对分子量为76kD的过敏原组分。     

Extraction,purification and properties of soil hydrolases

Invertase, cellulase, phosphatases, protease and β-glucosidase were extracted from permanent pasture soil with 0.2 M phosphate buffer (pH 8) in the presence of 0.2 M EDTA. This extract was further treated with ammonium and salmine sulphates. Attempts were made to fractionate these enzyme activities by gel and anion-exchange chromatography. Specific activities were estimated in all fractions and some characteristics of the purified enzymes (optimum pH, temperature and substrate concentration, and K_m and V_max) were investigated. The results indicated that extracted enzyme activities occurred partly in soil as a carbohydrate-enzyme complex and partly as a humo-carbohydrate complex.     

磷的吸附和表面电荷特征及其与华南地区某些土壤矿物的关系

The phosphate adsorption and surface charge characteristics of the tropical and subtropical soils derived from different parent materials in China were determined, and their relations to soil mineralogy were analysed. The results showed that all soil phosphate adsorption curves were well fitted by Freundlich equation and Langmuir equation. The maximum buffering capacity of P ranged from 66 to 9 880 mg kg^-1, with an increasing order of purple soil, skeletal soil, red soil, lateritic red soil, yellow soil and latosol; and the highest value was 149 times the lowest value, which indicated great differences among these soils in phosphate adsorption and supplying characteristics. The pH₀ (zero point of charge) values obtained by salt titration-potential titration varied from 3.03 to 5.49, and the highest value was found in the latosol derived from basalt whereas the lowest value was found in the purple soil. The correlation analysis indicated that the main minerals responsible for phosphate adsorption in the soils were gibbsite, amorphous iron oxide and kaolinite; and the pH₀ was mainly controlled by kaolinite, gibbsite and oxides.     

Comparative evaluation of Ca chloride and Ca phosphate for extractable sulfur in soils with a wide range in pH

Deficiency of sulfur (S) is becoming widespread in the rainfed systems of India, and there is increasing need for diagnosing the deficiency. Calcium chloride and Ca phosphate are commonly used for extracting available S in soils. Because of cost and the ease of availability locally, we prefer using Ca chloride as an extractant over Ca phosphate, for extracting available S. However, there is paucity of data on the comparative evaluation of the two extractants to extract available S, especially in soils having a wide range in natural pH (from acidic to alkaline range). It is recognized that soil pH plays a dominant role in the adsorption–desorption and extractability of sulfate‐S in soils. We compared the extraction of S by Ca chloride and Ca phosphate in 86 Indian soils having a wide range in pH (4.5 to 10.6). Sulfur in the extracts was determined by ICP‐AES. Considering all the 86 soil samples tested, there was an excellent agreement between the values of extractable S determined by using the two extractants (r = 0.96, p < 0.001). However, the correlation coefficient (r) between the values of extractable S by the two reagents, although highly significant, varied among the groups of soil samples according to the range in soil pH. The highest correlation coefficient (r = 0.99, p < 0.0001, n = 17) was found for soils with pH in the alkaline range (8.5–10.6), and the lowest correlation coefficient (r = 0.71, p < 0.0001, n = 58) was obtained with a set of soil samples with pH in the acidic range (4.5–6.5). For soil samples having pH in the near‐neutral range (6.7–7.3), an excellent agreement was observed (r = 0.93, p < 0.0001, n =11) between the extractable‐S values obtained by the two extractants. While Ca phosphate extracted higher amount of S compared to Ca chloride in soil samples with pH in the acidic range, the two extractants were equally effective for soil samples with pH in the neutral or alkaline range. Our results suggest that for most of the soils in the semiarid tropical regions, which have pH in the neutral to alkaline range, Ca chloride can replace Ca phosphate as an extractant for removing available S in such soils.