Bioaccumulation and toxicity of lead,influenced by edaphic factors: using earthworms to study the effect of Pb on ecological health

Purpose

Lead (Pb) is a highly studied contaminant with no known biological function that causes harmful adverse effects on ecological and human health. We tried to evaluate how protective the current soil regulatory levels are for Pb towards safeguarding the ecological health. In order to achieve this, our study evaluated the effect of soil texture and pH on the toxicity and availability of lead to earthworms in soils varying in soil properties.

Materials and methods

The earthworm Eisenia fetida was exposed to Pb in three soils with different physico-chemical characteristics. Pb solutions were homogenously mixed with soil to obtain concentrations ranging from 0 to 10,000 mg/kg Pb dry soil. Avoidance behaviour, weight loss and mortality were measured in this study to calculate the EC₅₀ and LC₅₀ values.

Results and discussion

Weight loss and mortality in earthworms due to Pb toxicity were in the following order: acidic > neutral > alkaline soil. The EC₅₀ values resulting in 50% decrease in worm weight over control for Pb in acidic, neutral and alkaline soils were 460, 3606 and 5753 mg/kg soil, respectively. Thus, the acidic soil recorded an EC₅₀ well below the soil guideline value for Pb. Whereas, the LC₅₀ values resulting in 50% mortality in worms over control were 1161, 4648 and 7851 mg/kg, respectively, for acidic, neutral and alkaline soils. The Pb concentrations in earthworms ranged from 0.2 to 740 mg/kg wet weight. Soils with low clay content and acidic to neutral pH values demonstrated an increased Pb toxicity in earthworms compared to the soils with alkaline pH.

Conclusions

The worm weight loss is a more sensitive parameter than the mortality. This study emphasizes that the soil regulatory levels for Pb are not protective of worms in acidic soils. Therefore, care should be taken when using the current regulatory limits to assess and predict the safety of a contaminated site with acidic soils towards the ecological health.

     

Development of QSAR's in soil ecotoxicology: Earthworm toxicity and soil sorption of chlorophenols,chlorobenzenes and chloroanilines

Soil adsorption and the toxicity of four chloroanilines for earthworms were investigated in two soil types. The toxicity tests were carried out with two earthworm species, Eisenia andrei and Lumbricus rubellus. LC₅₀ values in mg kg^?1 dry soil were recalculated towards molar concentrations in pore water using data from soil adsorption experiments. An attempt has been made to develop Quantitative Structure Activity Relationships (QSAR's) using these results and data on five chlorophenols and dichloroaniline in four soils and five chlorobenzenes in two soils published previously (Van Gestel and Ma, 1988, 1990; Van Gestel et al., 1991). Significant QSAR relationships were obtained between 1) adsorption coefficients (log K _om ) and the octanol/water partition coefficient (log k _ow ), and 2) LC₅₀ values (in itμmol L^?1 soil pore water) and log K _ow . It can be concluded that both earthworm species tested are equally sensitive to chlorobenzenes and chloroanilines, E. andrei is more sensitive than L. rubellus to chlorophenols.     

Forms of Al in soil and soil solution in a long‐term fertilizer application experiment

Under the conditions of a long‐term fertilizer experiment, this study aimed to determine the contents of total and exchangeable aluminium in soil as well as the Al concentration in the soil solution. Additionally, Al speciation was evaluated with the use of the MINTEQA2 software. The results obtained indicated that under the conditions of long‐term application of different mineral fertilizers or farmyard manure, the soil reaction changed to a great extent (pH 3.58–6.78). At the same time, the content of total Al in soil fluctuated from 18.85 to 22.13 g/kg and that of exchangeable Al ranged from 1.42 to 102.66 mg/kg. The concentration of Al in the soil solution was highly differentiated (5.19–124.07 μmol/L) as well as that of free aluminium ions (Al³⁺) (0–16.9 μmol/L). In acidic soils, aluminium complexes with organic matter are the predominant forms of Al in the soil solution. In soils with neutral soil reaction, there were no free aluminium ions. Soil liming and addition of organic amendment were the treatments that restricted the presence of toxic aluminium forms in soil.     

The effect of two endogeic earthworm species on zinc distribution and availability in artificial soil columns

The objective of this study was to determine the impact of earthworm bioturbation on the distribution and availability of zinc in the soil profile.Experiments were carried out with Allolobophora chlorotica and Aporrectodea caliginosa in 24 perspex columns (∅ 10 cm), filled with 20-23 cm non-polluted soil (OM 2%, clay 2.9%, pH 0.01 M CaCl₂ 6.4), that was covered by a 3-5 cm layer of aged zinc spiked soil (500 mg Zn/kg dry soil) and another 2 cm non-polluted soil on top. After 80 and 175 days, columns were sacrificed and each cm from the top down to a depth of 15 cm was sampled. Earthworm casts, placed on top of the soil, were collected. Each sample was analyzed for total and CaCl₂-exchangeable zinc concentrations.Effects of earthworm bioturbation were most pronounced after 175 days. For A. chlorotica, total and CaCl₂-exchangeable zinc concentrations in the polluted layers were lower with than without earthworms. Total zinc concentrations in the non-polluted layers were higher in columns with earthworms. Casts of A. chlorotica collected on the soil surface showed slightly higher total zinc concentrations than non-polluted soil. Casts were found throughout the whole column. For A. caliginosa there were no differences in total zinc concentration between columns with and without earthworms. CaCl₂-exchangeable zinc concentrations in the polluted layers were lower for columns with earthworms. Casts were mainly placed on top of the soil and contained total zinc concentrations intermediate between those in non-polluted and polluted soil layers.This study shows that different endogeic earthworm species have different effects on zinc distribution and availability in soils. A. chlorotica transfers soil throughout the whole column, effectively mixing it, while A. caliginosa decreases metal availability and transfers polluted soil to the soil surface.     

Behavior and respiration responses of the earthworm Eisenia fetida to soil arsenite pollution

Soil arsenic (As) pollution from mining and industrial sources is a serious issue in China. Earthworms are considered ecosystem engineers and contribute to soil fertility development and maintenance of soil physico-chemical properties. In this study, earthworms were exposed to soils with different sodium arsenite concentrations (0, 5, 20, and 80 mg As kg^-1) for 60 d to investigate the changes in soil properties and the responses of the earthworms (e.g., burrowing activity and respiration). Earthworm burrowing activity decreased with increasing arsenite concentrations, and earthworm respiration was significantly lower in soils with 20 and 80 mg As kg^-1 compared to 0 mg As kg^-1. Changes in soil properties were also observed after incubation of As-amended soil with earthworms. Specifically, soil pH decreased, while soil electrical conductivity and contents of soil NH₃^--N, Olsen-P, and available K increased. Our results suggest that arsenite negatively impacts the metabolic activity of earthworms, leading to reduced burrowing activity, which in turn modifies the effects of earthworms on soil fertility and remediation.     

Lethal and sublethal effects of imidacloprid on two earthworm species (Aporrectodea nocturna and Allolobophora icterica)

The chemical imidacloprid is the major component of many widely used insecticides and is relatively persistent in soils. A set of experiments was carried out to estimate the lethal (mortality) and sublethal (weight loss) effects of one of these insecticides, Confidor, on two earthworm species commonly found in agricultural soils. A preliminary experiment in the absence of earthworms showed that imidacloprid was not rapidly degraded, with a decrease of less than 10% after 2 weeks, and that it was distributed in a reasonably homogeneous manner throughout the soil (less than 10% of variation between samples). The LC₅₀ of imidacloprid for the anecic species Aporrectodea nocturna and the endogeic species Allolobophora icterica was between 2 and 4 mg kg^–1 dry soil. This result is consistent with previous findings obtained with other earthworm species and natural soils. When sublethal effects were examined, significant decreases in weight were observed at concentrations of 0.5 and 1 mg kg^–1 dry soil for the two earthworm species whereas no effect was observed at a concentration of 0.1 mg kg^–1 dry soil (NOEC value). These concentrations are close to 0.33 mg kg^–1 which is the Predictive Environmental Concentration. Weight loss appears to be a valuable endpoint that can be used with worms freshly collected in the field as long as variability in the response of a control is taken into account.     

Ecological implications of motor oil pollution: Earthworm survival and soil health

The nontarget effects of fresh and used motor oil were studied in a soil test system involving such criteria as earthworm survival, response of soil dehydrogenase and urease, and nitrification. When earthworms were exposed to motor oil-contaminated soil for 4 weeks, the observed median lethal concentrations (LC₅₀) were 40.33 and 3.88 g kg⁻¹ soil for fresh and used oil, respectively. Only fresh motor oil application increased earthworms' body weight even at the higher dose of 19 g kg⁻¹ soil. Gas chromatography/mass spectrometry revealed that used motor oil contained more of aromatic hydrocarbons and heavy metals than fresh oil. This disparity in the chemical composition might be the factor responsible for the significant toxicity of used motor oil towards earthworms. Activities of soil dehydrogenase and urease were significantly enhanced in presence of both the motor oils, while there was a significant inhibition in nitrification by the used motor oil even at a low concentration of 0.2 g kg⁻¹ soil. This study clearly demonstrated that earthworm survival and nitrification could serve as suitable indices to assess motor oil pollution in soil.     

Effects of Alum-treated Waste Water Sludge on Barley Growth

Alum sludge derived from a municipal wastewater plant was used as a soil amendment in a greenhouse study with barley (Hordeum vulgare) as the test crop. Treatment variables included the soil pH (4.5, 5.1 and 6.5), the amount of Al in the sludge (control = 30 mg Al_T/g; alum sludges = 38 and 52 mg Al_T/g), and the sludge application rate (100 and 270 kg N_T/ha). Soil amendment with the two alum sludges reduced soil pH, increased Al³⁺ activity in the soil solution, and reduced barley growth over the 6-week experiment. Barley growth decreased as the Al³⁺ activity in the sludged soil solution increased, but for a given Al³⁺ the phytotoxicity of Al was markedly pH dependent. For example, at a pH of 5.0 ± 0.1 an Al³⁺ activity of 0.5 μM was sufficient to inhibit plant growth by about 50% this IC₅₀ value increased five-fold to about 2.5 μM when the soil pH was 4.5 ± 0.1. This decrease in the toxicity of Al50 with acidification was explained in terms of a competitive interaction between the H⁺-ion and Al³⁺ at the root surface. Stepwise multiple regression allowed the prediction of aerial leaf biomass from soil pH and sludge application rate.     

Effects of endogeic earthworms on soil processes and plant growth in coniferous forest soil

Summary The effects of the endogeic earthworm, Aporrectodea caliginosa tuberculata (Eisen) on decomposition processes in moist coniferous forest soil were studied in the laboratory. The pH preference of this species and its effects on microbial activity, N and P mineralization, and the growth of birch seedlings were determined in separate pot experiments. Homogenized humus from a spruce stand was shown to be too acid for A. c. tuberculata. After liming, the earthworms thrived in the humus and their biomass increased (at pH above 4.8). In later experiments in which the humus was limed, the earthworms positively influenced the biological activity in humus and also increased the rate of N mineralization. A. c. tuberculata increased the growth of birch seedlings, with increases observed in stems, leaves, and roots. Neither NH ₄ ⁺ -N fertilizer nor mechanical mixing with artificial worms affected seedling growth. No plant-growth-affecting compounds (e.g., hormone-like compounds) due to the earthworms were present in the humus. The shoot: root ratio in the birch seedlings was not affected by either the earthworms or the fertilizer. The experiments revealed the impact of earthworm activity on soil processes and plant growth.     

The response of Aporrectodea rosea and Aporrectodea trapezoides (Oligochaeta: Lubricidae) to moisture gradients in three soil types in the laboratory

The question of whether the response of earthworms to soil moisture is governed by their reaction to soil wetness (moisture content) or to soil water energy (matric suction) was examined in two species of earthworm using moisture gradients in three contrasting soil types with clay contents varying from 4 to 39%. Gravimetric moisture gradients ranging over 5–30% were established in horizontal cores comprising 12 or 14 sections containing loosely packed soil. Earthworms were introduced to each section at the beginning of each experiment. The earthworms moved from sections containing dry soil into adjacent sections containing moister soil. Clear effects were evident after 6 h but these became more obvious after 96 h. For the earthworm Aporrectodea rosea, the threshold soil mositure level at which earthworms were induced to move away from dry soil was a matric suction of about 300 kPa (pF 3.4) and was independent of soil type. In contrast, for A. trapezoides, the threshold soil moisture varied with soil type (sandy loam 15 kPa, loam 25 kPa, clay 300 kPa). We conclude that, for the earthworm A. rosea, matric suction and not water content of soil provided the cue by which the earthworm recognized dry soil. For A. trapezoides, there was an interaction between matric suction and soil type in which the response of A. trapezoides to soil moisture varied with soil texture and the threshold for avoidance of dry soil ranged from a matric suction of 300 kPa (20% w/w) in clay to 15 kPa (10% w/w) in sandy loam.     

Earthworm colonisation of abandoned arable soil polluted by copper

The distribution, density and biomass of earthworms were investigated at the copper polluted site, Hygum (Denmark). In 1994, shortly after farming of the area was abandoned, only four earthworm species were present and their distribution was restricted to areas where copper concentration did not exceed 200 mg kg^?1 dry soil. Sixteen years later (in 2010), without any agricultural activity, ten species of earthworms were found, in particular, epigeic species were present where soil copper concentrations reached >1000 mg kg^?1 dry soil.     

Significance of earthworms in stimulating soil microbial activity

 The stimulatory effect of earthworms (Lumbricus terrestris L.) on soil microbial activity was studied under microcosm-controlled conditions. The hypothesis was tested that microbial stimulation observed in the presence of a soil invertebrate would be due to the utilization of additional nutritive substances (secretion and excretion products) that it provides. Changes in microbial activity were monitored by measuring simultaneously CO₂ release and protozoan population density. The increase in CO₂ released in the presence of earthworms was found to result from both earthworm respiration and enhanced microbial respiration. The stimulation of microbial activity was confirmed by a significant increase in protozoan population density, which was 3–19 times greater in the presence of earthworms. The respiratory rate of L. terrestris was estimated to be 53 μl O₂ g^–1 h^–1. Earthworm respiration significantly correlated with individual earthworm weight, but there was no correlation between the increase in microbial respiration and earthworm weight. This finding does not support the hypothesis given above that enhanced microbial respiration is due to utilization of earthworm excreta. A new hypothesis that relationships between microbial activity and earthworms are not based on trophic links alone but also on catalytic mechanisms is proposed and discussed. Received: 26 August 1997     

Influence of cadmium,copper, and pentachlorophenol on growth and sexual development of Eisenia andrei (Oligochaeta; Annelida)

Summary In the existing guidelines for earthworm toxicity testing, mortality is the only test criterion. Mortality is, however, not a very sensitive parameter, and from an ecological point of view growth and reproduction are more important for a proper risk assessment of chemicals in soil. In this study the growth and sexual development of juvenile earthworms were considered as test criteria in a standardized earthworm toxicity test. The effect of Cd, Cu, and pentachlorophenol on the growth and sexual development of juveniles of the species Eisenia andrei was studied in an artificial soil substrate. Two tests with Cd were carried out to study the effects of the mode of application of the food source (cow dung). EC₅₀ (50% effective concentration) values for the effect of Cd, Cu and pentachlorophenol on the growth of E. andrei were 33–96, >100, and >32 mg kg^-1 dry soil, respectively, and there was no observed effect at 18–32, 56, and 32 mg kg^-1 dry soil, respectively. Sexual development of the earthworms was inhibited at 10 mg Cd kg^-1 and 100 mg Cu kg^-1 dry soil, but was not affected at the highest pentachlorophenol concentration tested (32 mg kg^-1 dry soil). The results were the same whether the food was applied in a hole in the middle of the soil or mixed homogeneously through the soil.     

Increases in pH and soluble salts influence the effect that additions of organic residues have on concentrations of exchangeable and soil solution aluminium

It has been suggested that additions of organic residues to acid soils can ameliorate Al toxicity. For this reason the effects of additions of four organic residues to an acid soil on pH and exchangeable and soil solution Al were investigated. The residues were grass, household compost, filter cake (a waste product from sugar mills) and poultry manure, and they were added at rates equivalent to 10 and 20 t ha^?1. Additions of residues increased soil pH measured in KCl (pH_(KCl)) and decreased exchangeable Al³⁺ in the order poultry manure > filter cake > household compost > grass. The mechanism responsible for the increase in pH differed for the different residues. Poultry manure treatment resulted in lower soil pH measured in water (pH_(water)) and larger concentrations of total (Al_T) and monomeric (Al_mono) Al in soil solution than did filter cake. This was attributed to a soluble salt effect, originating from the large cation content of poultry manure, displacing exchangeable Al³⁺ and H⁺ back into soil solution. The considerably larger concentrations of soluble C in soil solution originating from the poultry manure may also have maintained greater concentrations of Al in soluble complexed form. There was a significant negative correlation (r = ?0.94) between pH_(KCl) and exchangeable Al. Concentrations of Al_T and Al_mono in soil solution were not closely related with pH or exchangeable Al. The results suggest that although additions of organic residues can increase soil pH and decrease Al solubility, increases in soluble salt and soluble C concentrations in soil solution can substantially modify these effects.     

Effects of earthworms on decomposition and metal availability in contaminated soil: Microcosm studies of populations with different exposure histories

Population-specific differences in the responses of earthworms to simultaneous exposure to Cu and Zn were studied in microcosm experiments. Two populations of Aporrectodea caliginosa tuberculata (Eisen) with different metal exposure histories were chosen for the studies. Microcosms were prepared containing either uncontaminated soil or soils with low or high combined Cu/Zn -concentrations (79/139 or 178/311 mg kg⁻¹ dry mass of soil, respectively). Earthworms from each population were introduced to the microcosm treatments with some microcosms serving as controls without earthworms. One series of microcosms was destructively sampled after 16 weeks incubation in a climate chamber. Survival, growth, reproduction and decomposition by earthworms in each treatment were measured. An additional microcosm series was sampled for soil and earthworm measurements at four weeks intervals to determine temporal changes in the availability of metals in the soils and their accumulation into earthworms. Cu and Zn were sequentially extracted from the soil samples of both microcosm series to estimate mobility and availability of the metals in the soil. Earthworms with long-term exposure history to metal-contaminated soil seemed to tolerate higher soil metal concentrations than earthworms without earlier exposure. Both earthworms and metals affected soil respiration (CO₂ production) and nitrogen mineralization. In addition, earthworms seemed to decrease the mobility and bioavailability of metals in the soil through their burrowing activity.     

The influence of soil characteristics on the toxicity of four chemicals to the earthworm Eisenia fetida andrei (Oligochaeta)

Summary The acute toxicity of Cd (chloride), chloroacetamide, 3,4-dichloroaniline and pentachlorophenol to the earthworm Eisenia fetida andrei was determined using the OECD (1984) artificial soil and contact testing procedures. To investigate the influence of two soil characteristics (pH and organic-matter content), the toxicity of the chemicals was also determined in two natural sandy soils. It is concluded that the filter-paper contact test cannot be recommended to predict earthworm toxicity of these chemicals in soil. Toxicity in soil was influenced by both pH and organic-matter content. Differences between LC₅₀ values in the high-organic-matter artificial soil and in an acid, low-organic-matter sandy soil were, however, not greater than a factor of 3–4. The results of this study therefore support the use of a well-defined artificial soil substrate for standardized earthworm toxicity tests.     

Earthworm bioturbation stabilizes carbon in non-flooded paddy soil at the risk of increasing methane emissions under wet soil conditions

Studies on earthworms in rice-based ecosystems tend to focus on some pest species, while the potential of these important soil engineers for beneficially affecting carbon storage and cycling is widely ignored. We carried out a microcosm experiment to quantify the impact of the tropical earthworm Pheretima sp. on the C turnover in paddy soils under different conditions of water saturation and N fertilization. The soil was sampled at the lowland farm of the International Rice Research Institute (Philippines). In the absence of earthworms, soil respiration showed a distinct hump-shaped maximum at intermediate levels of water saturation (4-fold higher than in hand-dry soil) and increased 1.5-fold with increasing amounts of N fertilization. Amounts of CH₄ emitted, in contrast, were small at low to moderate soil humidity and became very high under conditions of water saturation (80-fold higher than hand-dry soil). No response to nitrogen addition was observed. Earthworms suppressed both the respiration maximum at intermediate saturation levels (by a factor of 1.4) and the stimulating impact of N fertilization (1.7-fold at maximum fertilizer level). On the other hand, earthworms strongly increased CH₄ release under conditions of high water saturation (3-fold). No consistent response of the soil microflora (bacterial abundance, soil enzymes) to earthworm activity could be established. Our findings suggest that the stabilization of soil organic C via earthworm bioturbation is confined to the range of soil humidity that allows high activity of Pheretima sp. Under conditions of intensive agriculture, the stabilizing effect of the worms may even be augmented by the fact that they offset the positive effect of N fertilization on microbial respiration. Earthworms may thus play a vital role in reducing the CO₂ flush from paddy soils after the conversion to non-flooded crops such as aerobic rice or maize. Acceleration of methane emission in very humid soils nevertheless points to a certain risk that is associated with increasing earthworm abundance in production systems that are still exposed to temporary flooding during the wet season.     

不同磷水平对红壤溶液中锰、铝、镁和钙浓度变化以及小麦生长的影响

为了探明不同磷水平对红壤中土壤溶液主要金属离子变化的影响以及小麦对磷的响应,确定红壤中小麦适宜的施磷水平,采用原位提取土壤溶液和比较生物量的方法,监测了短期内红壤溶液中主要金属离子浓度变化及小麦生物量的变化。结果表明:碳酸钙的加入可以显著升高酸性红壤的p H,土壤溶液中铝、锰和镁浓度显著低于未加碳酸钙处理;800 mg/kg磷处理后铝、锰、镁和钙的浓度要比未加磷处理分别至少降低47%、44%、37%和33%。随着施磷量的增加,小麦在200 mg/kg磷处理时积累的生物量最大,随后磷增加,小麦生物量反而降低。而加碳酸钙处理小麦地下部生物量随着施磷量增加则降低。结果表明碳酸钙不仅可以有效升高土壤p H,降低土壤溶液铝浓度,还降低土壤溶液中锰的浓度。磷的加入同样可以降低锰和铝的浓度,缓解铝和锰毒害。红壤中生长小麦的适宜施磷量为200 mg/kg。     

Life-history traits and population growth rate in the laboratory of the earthworm Dendrobaena octaedra cultured in copper-contaminated soil

A study on the widespread earthworm Dendrobaena octaedra was conducted to determine which individual life history traits were the most sensitive to copper and to determine the contribution of changes in individual traits to changes in the population growth rate (λ). The study showed that the effect of copper on population growth rate mirrored the effects seen on growth, maturation and reproductive output, with stimulation at the lowest concentrations and inhibition at the highest concentration. A decomposition analysis showed that the mean change in λ was mainly driven by time between consecutive cocoon productions, except at the highest copper concentration (200 mg/kg dry soil) where decreased production of fertile cocoons also contributed to the reductions in λ. The highest population growth rate (λ = 1.18 week⁻¹) occurred at 80 mg Cu/kg dry soil. At higher concentrations λ became gradually smaller, and was almost 1 week⁻¹ (where no population increase or decrease occurs) at the highest exposure concentration of 200 mg Cu/kg dry soil suggesting that extinction would occur if a population of D. octaedra were to be exposed to copper concentrations only slightly higher than this level.     

Differential tolerances of two old world bluestem genotypes to excess aluminum in acid soil and nutrient solution

Two genotypes of Old world bluestems from the species Bothriochloa intermedia (R. Br.), A. Camus, shown earlier to differ in tolerance to acid, Al‐toxic Tatum subsoil at pH 4.1, were characterized further with respect to growth in pots of Tatum soil over a wider pH range and tolerance to Al in nutrient solutions. The two genotypes studied were acid‐soil tolerant P. I. 300860 (860) and acid soil sensitive P. I. 300822 (822).

The soil experiment confirmed earlier rankings of acid soil tolerance in these two genotypes. For example, with 0, 375 or 750 ug CaCO₃ g^‐1 soil (final pH 4.0, 4.3 and 4.6), the 860 genotype produced significantly more dry top weight than 822, but these differences were precluded with 1500 or 3000 ug g^‐1 CaCO₃ added (pH 4.7 and 5.4). At pH 4.3 and 4.6, the root dry weights of the two genotypes were also significantly different and weights were equalized at pH 4.7 and 5.4. The 860 genotype made fairly good top growth (67% of maximum) at pH 4.3 and a soil Al saturation of 63%; this situation was lethal for 822. When grown in greenhouse pots, the acid‐soil tolerant 860 genotype required only about one fourth as much CaCO₃ as 822 to produce good growth of forage on acid Tatum subsoil. If confirmed under field conditions, such a difference could be economically significant in reclaiming acidic marginal land and in producing forage at low cost.

Differential Al tolerance in the two genotypes was confirmed in nutrient solutions. For example, with 8 mg Al L^‐1 added, both top and root dry weights of 860 were significantly higher than those of 822, but with no Al added, these growth differences disappeared.

Mineral analyses of plants did not shed much light on mechanisms of differential acid soil or Al tolerance. For example, Al concentrations in plant tops associated with toxicity varied from 33–43 ug g^‐1 in nutrient solutions containing Al to 119–283 ug g^‐1 in acid soil It appears that elucidation of Al‐adaptive mechanisms will require physiological and biochemical studies at the cellular level.