Identification and Characterization of High Acid Tolerant and Aluminum Resistant Yeasts Isolated from Tea Soils

From acidic tea soils of Kagoshima Prefecture in Japan, some soil properties were determined and 38 strains of acid tolerant microorganisms were isolated. Different Al³⁺ concentrations were applied to YG media to estimate Al resistance. Selected microbial strains could grow strongly in the liquid media in the presence of 100 mM Al³⁺ and survive even in 300 mM Al³⁺ at pH 3.0. Their base sequences of 28S rDNA-D1/D2 were determined and sequence data were searched using the Basic Local Alignment Search Tool (BLAST) system. The results of sequencing revealed that the isolates belong to two different species, Cryptococcus sp. and Candida palmioleophila. When cultivated with various Al³⁺ concentrations, the yeast growth was inhibited at a concentration of 200 mM. Pre-cultivation of these strains with 0–30 mM Al³⁺ did not promote the growth response caused by Al³⁺. Inductively-Coupled Plasma-Mass Spectrometry (ICP-MS) was used to assess the elimination of Al. The amount of Al remaining in culture media was decreased considerably after cultivation. Due to a capacity for resistance to significant Al concentrations as well as high Al elimination, these acid tolerant and Al resistant yeasts may have potential applications in the bio- and phyto-remediation of Al and acid-contaminated soils.     

Identification and Characterization of Yeasts with Tolerance to High Acidity and Resistance to Aluminum Isolated from Tea Soils

From acidic tea soils in Kagoshima Prefecture in Japan, 38 strains of acid-tolerant microorganisms were isolated and some soil properties were determined. Different Al concentrations were applied to YG media to estimate the Al resistance. Selected microbial strains were able to grow actively in liquid media in the presence of 100 mM Al and to survive even in 300 mM Al at pH 3.0. The base sequences of 28S rDNA-D1/D2 were determined and sequence data were searched using the Basic Local Alignment Search Tool (BLAST) system. The results of sequencing revealed that the isolates belonged to two different species, Cryptococcus sp. and Candida palmioleophila . Subsequently, Neighbor-Joining molecular phylogenetic trees were constructed. When the yeasts were cultured at various Al concentrations, the growth was inhibited at a concentration of 200 mM. Pre-culture of these strains with 0–30 mM Al did not promote the growth response induced by Al. Inductively coupled plasma-mass spectrometry (ICP-MS) was used to assess the elimination of Al from YG media. The amount of Al remaining in the culture media decreased considerably after culture. Due to a capacity for resistance to significant Al concentrations as well as high Al elimination, these acid-tolerant and Al-resistant yeasts may offer potential applications for the bio- and phyto-remediation of acid soils with a high level of Al.     

Aluminium speciation and pH of an acid soil in the presence of fluoride

The aim was to determine whether the addition of F to an acid soil reduces the concentration of free Al³⁺ and other forms that have been shown to be toxic to plants. The ability of two different extracts to reflect Al speciation in the soil solution was also investigated. Addition of F (0-5.2μmolg⁻¹) to an acid soil (pH 4.15, soil solution) increased the pH and total concentrations of Al and F in the soil solution whereas Al³⁺ remained constant or decreased. Soil solution pH, total soluble Al and Al extracted by 0.01 m CaCl₂ are not good predictors of the likelihood of aluminium toxicity in soils containing soluble fluoride.     

Ca2+ Effects on K+ Fluxes in Arabidopsis Seedlings Exposed to Al3+

Potassium transport was investigated in the root elongation zone of Arabidopsis seedlings during the first minutes of Al³⁺ exposure, using the non-invasive MIFE microelectrode technique. To prevent pH changes during Al³⁺ application, and to separate aluminium from acidic stress, plants were pre-treated with 5 mM homoPIPES before addition of AlCl₃ (pH 4.2). The 30-min treatment with 50 or 500 μM AlCl₃ led to a significant increase in K⁺ efflux in solutions containing 100 μM CaCl₂. This efflux was suppressed by high concentrations of Ca²⁺ (10 mM) in the bathing solution. Our results suggest that elevated external Ca²⁺ activities can sustain K⁺ influx in the root elongation zone during Al³⁺ exposure either by maintaining [Ca²⁺]_cyt or by affecting Al³⁺ uptake across the plasma membrane.     

Aluminium solubility related to secondary solid phases in upper B horizons with spodic characteristics

We examined the aluminium solubility in the upper B horizon of podzols and its relation to the solid phase of the soil in 60 samples covering a pH range from 3.8 to 5.1. Solid phases were characterized by extractions with acid oxalate and pyrophosphate (pH 10). The solubility of Al was studied in a batch experiment in which samples were equilibrated with 1 m m NaCl at 8°C for 5 days. We also monitored the dissolution kinetics of Al and Si, in some samples. The oxalate and pyrophosphate extractions suggested that secondary Al was mainly organically bound in most soils, and imogolite-type materials seemed to constitute much of inorganic secondary Al. No single gibbsite or imogolite equilibrium could explain Al³⁺ activities. In all samples Al solubility, defined as log{Al³⁺} + 1.65pH, was closely related to the molar ratio of aluminium to carbon in the pyrophosphate extracts (Al_p/C_p). Solubility increased with the Al_p/C_p ratio until the latter reached ≈ 0.1. This indicated that solubility was controlled by organic complexation, at least when Al_p/C_p was small. Silica dissolved slowly in most soils used in the kinetic experiments. We conclude that imogolite-type materials in the upper B horizon dissolved slowly because of coating with humic substances or ageing or both.     

Prolonged leaching of mineral forest soils with dilute HCl solutions: the solubility of Al and soil organic matter

Long-term acidification has been shown to result in a considerable decrease in the amount of organically bound soil Al and in a gradual decrease in the solubility of Al. We examined the solubility of soil organic matter (SOM) and Al in four acid mineral soils (one Arenosol Ah, two Podzol Bh, and one Podzol Bs) as they were leached sequentially using a solution containing 0.001 m HCl and 0.01 m KCl. The acid leaching resulted in relative decreases in Al that were 2–6 times greater than for organic C. The organic C and Al dissolved by the acid leaching originated mainly in the pyrophosphate-extractable fraction of the elements. Protonation seems to be a major mechanism in stabilizing the residual SOM, as indicated by small changes in effective cation exchange capacity with the degree of acid leaching. In the samples of Podzol Bh and Arenosol Ah soils the solubility of Al (defined as log₁₀{Al³⁺} + 1.5pH) in equilibrium suspensions (0.01 m KCl) was closely related to the ratio of pyrophosphate-extractable Al to pyrophosphate-extractable organic C. The Podzol Bs sample probably contained a small amount of a surface-reactive Al(OH)₃ phase, which rapidly became depleted by the acid leaching.     

The solution chemistry of podzolic soils from the eastern Canadian shield: a thermodynamic interpretation of the mineral phases controlling soluble Al3+andH4SiO4

Thirty-one soil solutions were extracted by immiscible displacement with CCl₄ under high speed centrifugation from sub-horizons of three podzolic soils from north-eastern Ontario, Canada. The solutions were analysed for major cations and anions and a speciation of dissolved Fe and Al was attempted to distinguish 'free', 'organically bound' and 'inorganically bound' species. Results indicated that the Ae (E) horizon solutions were of low pH and contained mainly organically bound Fe and Al. With depth, _pHs increased, ionic strengths decreased and the relative proportion of inorganically bound Fe and Al increased. Although application of phase diagrams permitted only a semi-quantitative interpretation of the data, all horizon solutions, with the exception of some Ae solutions, appeared supersaturated with respect to likely occurring crystalline and amorphous aluminosilicates [kaolinite, halloysite, allophane (Al:Si=l) and imogolite]. Of the phases considered, reactions involving imogolite-allophane, gibbsite-halloysite, gibbsite-allophane and gibbsite-imogolite all appeared reasonable in controlling the content of Al³⁺ and H₄SiO₄ in solution, although the presence of gibbsite and imogolite could not be definitely confirmed in these soils.     

Geologic Nitrogen as a Source of Soil Acidity

The origin of highly acidic (pH<4.5) barren soils in the Klamath Mountains of northern California was examined. Soil parent material was mica schist that contained an average of 2,700 mg N kg⁻¹, which corresponds to 7.1 Mg N ha⁻¹ contained in a 10-cm thickness of bedrock. In situ soil solutions were dominated by H⁺, labile-monomeric Al³⁺ and NO₃⁻, indicating that the barren area soils were nitrogen saturated—more mineral nitrogen available than required by biota. Leaching of excess NO₃⁻ has resulted in removal of nutrient cations and soil acidification. Nitrogen release rates from organic matter free soil ranged from 0.0163 to 0.0321 mg N kg⁻¹ d⁻¹. Nitrogen release rate from fresh ground rock was 0.0465 mg N kg⁻¹ d⁻¹. This study demonstrates that geologic nitrogen may represent a large and reactive nitrogen pool that can contribute significantly to soil acidification.     

Effects of metals on the germination and growth of fungal isolates from New Caledonian ultramafic soils

Soils of New Caledonian mining areas have relatively high metal contents. Two ultramafic soils rich in extractable Mn, Fe, Ni and Co and two agricultural soils with much lower concentrations of metals were compared through different experiments. Microbial population numbers and their tolerance to Mn²⁺ and Ni²⁺ were estimated. The effect of five metals (Ni²⁺, Co²⁺, Fe²⁺, Mn²⁺ and Mg²⁺) upon spore germination and mycelial growth was investigated for fungi isolated from the two types of soils. Nickel appeared to be the most toxic metal, but the other metals also had inhibitory effects at concentrations similar to those which occur in ultramafic soils. Spore germination was more sensitive to metals than mycelial growth. The toxic effects of the five metals were not cumulative when mixed together into the medium; antagonism between ions, partly neutralizing their effects may account for this. Fungal isolates from ultramafic soils appeared to be more tolerant to metals than those from agricultural soils. Some ultramafic soil isolates appeared to actually require the presence of metals, with their growth being stimulated by relatively high concentrations of these elements. Two fungal isolates of genera which are often common in agricultural soils (i.e. Curvularia and Fusarium), and which are not present in New Caledonian ultramafic soils were also tested. Curvularia was more sensitive to the five metals than all the other isolates and Fusarium was particularly inhibited by Ni²⁺. However, a proportion of the spores of each isolate was able to germinate and to grow moderately well in the presence of relatively high metal concentrations. Their absence from ultramafic soils is thought to result from their elimination by competitors that are more tolerant to metals.     

Microbial biomass and nitrogen transformations in surface soils strongly acidified by volcanic hydrogen sulfide deposition in Osorezan, Japan

Volcanic acidification has created unique ecosystems that have had to adapt to the acidic environments in volcanic regions. To characterize the primary microbial properties of strongly acidified soils in such environments, we investigated microbial biomass, nitrogen transformations and other relevant chemical properties in the surface soils of solfatara and forests from Osorezan, a typical volcanic region in Japan, and compared the results to common Japanese forest soils. Soil microbial biomass C (MBC) and N (MBN) were determined using the chloroform fumigation–extraction method. Potential net N mineralization and net nitrification were measured in aerobic laboratory incubations. Long-term acidification in the Osorezan soils by volcanic hydrogen sulfide deposition caused low soil pH (3.0–3.8), base cation deficiency and increased concentrations of toxic ions such as Al³⁺. The proportions of MBC to total carbon (MBC/TC ratio) and MBN to total nitrogen (MBN/TN ratio) were lower than those in common Japanese forest soils. The extreme acidic conditions may have inhibited microbial survival in the Osorezan acid soils. Net N mineralization occurred at rates comparable to those in common Cryptomeria japonica forest soils, probably because of the presence of acid-tolerant soil microorganisms. Net nitrification was completely inhibited and autotrophic ammonia oxidizers were not detected by the MPN method. The inhibition of nitrification prevents nitrogen leaching from the soils, thus maintaining a nitrogen cycle in the volcanic acid region in which     (and NH₃) is recycled among microorganisms and plants.     

Inhibition of nitrate accumulation in tropical grassland soils: effect of nitrogen fertilization and soil disturbance

In acid soils in the Eastern Plains of Colombia, forage grasses planted on land prepared before the previous dry season produced 40–50% more dry matter than when land was prepared immediately before planting. Virtually no NO₃⁻ accumulated in surface (0–10 cm) soil from three native undisturbed savanna sites. Where land was ploughed before the dry season, NO₃⁻ levels increased gradually after a 2–3 month lag, and dropped at the beginning of the rains. In samples incubated for 4 weeks, more NO₃⁻ accumulated in the wet than the dry season. A similar 2–3-month lag occurred when land was ploughed after the dry season. NH₄⁺ levels were higher in ploughed than savanna soils, and rose in all soils at the beginning of the rains. More NO₃⁻ and NH₄⁺ accumulated on incubation in pots than in soil cores. Forage grasses inhibited NO₃⁻ accumulation in the soil, relative to plant-free plots, and legumes stimulated it. N fertilization overcame this inhibition except in the case of Brachiaria humidicola .     

Fixation of radiocaesium traces in a weathering sequence mica → vermiculite → hydroxy interlayered vermiculite

Radiocaesium fixation in soils is reported to occur on frayed edge sites of micaceous minerals. The weathering of mica in acid soils may therefore influence the Cs⁺ fixation process and thereby the mobility of the radiopollutant. We produced a laboratory weathering model biotite → trioctahedral vermiculite → oxidized vermiculite → hydroxy interlayered vermiculite (HIV) and quantified the Cs⁺ fixation of each mineral both in a fixed K⁺–Ca²⁺ background and in acid conditions. The transformation process was achieved through K depletion by Na-tetraphenylboron, oxidation with Br₂ and Al-intercalation using NaOH and AlCl₃. In a constant K⁺–Ca²⁺ background, vermiculite fixed 92–95% of the initial ¹³⁷Cs⁺ contamination while biotite and HIV fixed only 18–33%. In acid conditions, the interlayer occupancy by either potassium (biotite) or hydroxy-Al groups (HIV) strongly limited Cs⁺ fixation to 1–4% of the initial ¹³⁷Cs⁺ contamination. Cs⁺ fixation occurred on vermiculitic sites associated with micaceous wedge zones. Though both oxidized and trioctahedral vermiculites fixed similar Cs⁺ amounts in a constant K⁺–Ca²⁺ background (92–95%), the oxidized vermiculite retained much more radiocaesium in acid conditions (78–84% against 54–59%), because of its dioctahedral character.     

Metal-humus complexes in A horizons of Thai and Korean red and yellow soils

Carbon, Al and Fe (C_pyr, Al_pyr and Fe_pyr) were extracted with 0.1 m Na₄P₂O₇ from 26 A horizon samples of tropical Thai and temperate Korean soils (Ultisols, Alfisols, Oxisols and Inceptisols). The soils, except for one Thai Inceptisol, had similar total C (0.35–3.29%) and C_pyr/total C ratios (0.20–0.41). There were approximately linear relationships between total C or C_pyr and clay content; two groups of soils gave different linear relationships. A curvilinear relationship between C_pyr and (Al + Fe)_pyr (milli-atom kg⁻¹) that can be approximated by an equation: C_pyr= 53 (Al_pyr+ Fe_pyr)^1/2– 24 was also found for most Thai and Korean soils. The above relationships indicated that total C and C_pyr would be close to zero at zero clay or zero (Al + Fe)_pyr. It was inferred that clay-humus interaction has a primary importance in the determination of humus content in red and yellow soils in tropical and temperate regions and that the main role of clay is to supply Al and Fe that complex and stabilize humus against microbial degradation.     

Correlations between extractable Na, K, Mg, Ca, P & N from fresh and dried samples of two Aquults

Significant increases in extractable ions resulted from air-drying and grinding samples of two infertile Aquults. Effects of the sample preparation differed markedly between ions and between the two soils. Regression equations were calculated to predict extractable ions in dried, ground samples from extractable ions in fresh, unground samples and the relationships were compared between the two soil series. Regressions were significantly different between soils for extractable PO³₄, Mg⁺⁺, and K⁺, but not for Ca⁺⁺ and Na⁺. Extractable NH ⁺₄ and NO-₃ in fresh, unground samples were not correlated with those in air-dry, ground samples of either soil. Differences in response to preparation between soil types appeared to be related to the oxidative status of these soils in the field, wherein constituents of more poorly-drained soils may be less stable to the oxidizing conditions of air-drying and grinding. Such complexities suggest that effects of sample preparation should be considered when interpreting soil nutrient data for studies of forest nutrient cycling and forest soil fertility.     

Soil acidity and aluminium toxicity as affected by surface liming and cover oat residues under a no-till system

A 3-year field trial examined in a long-term no-till system the effects of surface-applied lime and cover black oat ( Avena strigosa Schreb) residues on soil chemical attributes, root growth and grain yield of corn ( Zea mays L.) and soybean ( Glycine max L. Merrill) on a loamy, kaolinitic, thermic Typic Hapludox in Paraná State, Brazil. The treatments consisted of dolomitic lime broadcast on the soil surface at 0 or 12 t/ha, with and without cover of black oat residues. Corn and soybeans were grown without rainfall limitation. Applying lime on the surface improved soil acidity and decreased aluminium (Al) toxicity to a 10-cm depth 1 year after application. Surface liming increased pH and the content of exchangeable Ca²⁺ to a 20-cm depth, and decreased Al toxicity to a 40- to 60-cm depth, 3 years after application, indicating that the surface-applied lime moved deeper. Cover black oat residues did not favour the mobility of surface-applied lime to alleviate subsoil acidity and an increase in the Al³⁺ saturation level at the soil surface was found in unlimed plots with black oat residues. Root growth and grain yields of corn and soybean were not influenced by surface liming with or without cover black oat residue. Despite the soil acidity level, root length of corn and soybean ranged from 55 to 60% at 0- to 10-cm depth. The results suggest that Al toxicity is low in no-till systems during cropping seasons with adequate and well-distributed rainfall, but this effect is not related to the presence of cover oat residues.     

Aluminum tolerance associated with enhancement of plasma membrane H+-ATPase in the root apex of soybean

Seventeen soybean cultivars were screened to discern differences in aluminum (Al) sensitivity. The Sowon (Al-tolerant) and Poongsan (Al-sensitive) cultivars were selected for further study by simple growth measurement. Aluminum-induced root growth inhibition was significantly higher in the Poongsan cultivar than in the Sowon cultivar, although the differences depended on the Al concentration (0, 25, 50, 75 or 100 μmol L^–1) and the amount of exposure (0, 3, 6, 12 or 24 h). Damage occurred preferentially in the root apex. High-sensitivity growth measurements using India ink implicated the central elongation zone located 2–3 mm from the root apex. The Al content was lower 0–5 mm from the root apices in the Sowon cultivar than in the apices of the Poongsan cultivar when exposed to 50 μmol L^–1 Al for 12 h. Furthermore, the citric acid exudation rate was more than twofold higher in the Sowon cultivar. Protein production of plasma membrane (PM) H⁺-ATPase from the root apices (0–5 mm) was upregulated in the presence of Al for 24 h in both cultivars. This activity, however, decreased in both cultivars treated with Al and the Poongsan cultivar was more severely affected. We propose that Al-induced growth inhibition is correlated with changes in PM H⁺-ATPase activity, which is linked to the exudation of citric acid in the root apex.     

Analysis of Potassium Uptake by Rice Roots Treated with Aluminum Using a Positron Emitting Nuclide, 38K

We investigate the effect of Al on K⁺ uptake by rice roots. Potassium-38 (³⁸K), a positron emitting nuclide (the half-life: 7.61 min), was used to trace K⁺ behavior. When a rice root was treated with 10μM Al for 24 h, the uptake of ³⁸K in the root was increased in the range of 1 to 2 cm from the root tip compared with that of the control sample. Because the root continued to grow without showing any damage of plasma membrane during the Al treatment, it was suggested that the ³⁸K uptake was not occurred through diffusion into the cells. The uptake of ³⁸K in both treatments, with/without Al, was decreased by VO₄^3- (inhibitor of H⁺-ATPase on plasma membrane) and DNP (H⁺ ionophore) treatment, which suggested that the K⁺ uptake was performed through an active transport, such as H⁺:K⁺ transport or H⁺ gradient promoted by an Al treatment.     

The factors limiting crop production on the Mubangwe Farm, Malawi

Abstract. Mubangwe Farm, northern Malawi, was brought into cultivation from savanna vegetation with Brachystegia between 1970 and 1983 to produce tobacco, maize and groundnuts. Because of poor yields even with inputs of lime, N, P and K the fields were soon abandoned. The soils were only moderately acidic with little likelihood of Al toxicity. They held only small amounts of exchangeable Ca²⁺ (down to 0.01 cmol_c/kg) particularly at depth. Phosphorus availability was low particularly in the subsoil. A minus-one pot experiment showed that the growth of sorghum in topsoil samples was limited by lack of N, P, S and Ca. The lower availabilities of P and Ca in the subsoil are therefore likely to be major limitations to growth, allowing only limited root penetration. The loamy sand to sandy clay soils generally had small available water capacities (down to 0.07 cm³/cm³) which, with restricted root development, may have lead to drought in dry periods during the growing season. The findings emphasize the need to measure both subsoil and topsoil properties when new areas of land are being developed for crop production.     

Rice-soil interactions in Vietnamese acid sulphate soils: impacts of submergence depth on soil solution chemistry and yields

Abstract. The aim of this study was to investigate the effects of water submergence depth on radial oxygen loss (ROL), soil solution chemistry and rice growth performance in acid sulphate soils in southern Vietnam. ROL was measured in a solution culture. In a separate pot experiment the impact of water submergence depth on rice growth and soil solution chemistry was studied. Three submergence depths were used in the two experiments (5, 10 and 15 cm). ROL declined with submergence depth and was significantly greater in young roots (with no root hairs) than in older roots. In the pot experiment rice growth and soil solution chemistry were clearly affected by the submergence depth. During the first crop at 5 cm submergence, there was a significantly higher yield and a higher oxidation state (pe+pH) compared to 10 or 15 cm submergence. The Fe concentration was significantly greater at the 5 cm depth compared to the 10 or 15 cm depth. SO₄^2– reduction was delayed at the 5 cm depth. Rice yield was c. 25% less at the 15 cm than at the 5 cm depth. During a second crop, there was a substantial SO₄^2- reduction and H₂S formation and almost no significant effects of submergence depth on either soil solution chemistry or crop yield. In a field experiment with a dry-season rice crop, yield and Fe, Al and SO₄^2– concentrations were higher at a shallow submergence depth than at greater depths in the same field, showing similar depth trends to those found during the first crop in the pot experiment. Farmers should be advised to use a shallow submergence depth and, if possible, avoid deep-rooted rice varieties. A conceptual model is suggested, which summarizes the relationships between ROL and soil solution chemistry.     

Influence of the nature of clay minerals on the fixation of radiocaesium traces in an acid brown earth–podzol weathering sequence

The magnitude of radiocaesium fixation by micaceous clay minerals is affected by their transformation, which depends on weathering in soil. The net retention of radiocaesium traces was quantified by sorption–desorption experiments in the various horizons of four sandy soils forming an acid brown earth–podzol weathering sequence derived from sandy sediments and characterized by marked changes in mineral composition. The features of the 2:1 minerals of the four soils, resulting from an aluminization process in depth and a desaluminization process towards the surface, had a strong influence on Cs⁺ fixation. Beneath the desaluminization front, which deepens from the acid brown earth to the podzol, hydroxy interlayered vermiculite was dominant and the ¹³⁷Cs⁺ fixation was the weakest. At the desaluminization front depth, vermiculite was responsible for the strongest ¹³⁷Cs⁺ fixation. In the upper layers, smectite appeared in the podzolized soils and the ¹³⁷Cs⁺ fixation decreased. The magnitude in Cs⁺ fixation therefore appeared as a tracer of the transformation process affecting the 2:1 clay minerals in the acid brown earth–podzol weathering sequence. This magnitude was positively correlated with the vermiculite content of the studied soil materials estimated by the rubidium saturation method.