Comparative study on the responses of broiler chicken to hot and humid environment supplemented with different dietary levels and sources of selenium

Present study was carried out with the objective of investigating the role of green synthesized nano Se (GNS) in growth performance, digestibility of minerals, immunity, stress alleviation, antioxidant status, and body Se content of broiler chicken raised under hot and humid environment with respect to market nano Se (MNS) and inorganic Se. The experimental design was 3 × 3 factorial, in which three levels (0.15, 0.20, and 0.25 ppm) and three sources (inorganic, green nano, and market nano) of Se resulted in nine treatments viz. IS-0.15, GNS-0.15, MNS-0.15, IZ-0.20, GNS-0.20, MNS-0.20, IS-0.25, GNS-0.25, and MNS-0.25 (IS: inorganic Se, GNS: green nano Se, MNS: market nano Se). A total of 432 broiler chicken were divided among nine treatments with six replicates of birds per treatment (8 birds/replicate). Results of present study revealed significantly better growth performance of birds supplemented with 0.25 ppm nano Se. The supplementation of 0.25 ppm nano Se improved the immune response and lymphoid organ development of birds. Significantly higher Se and nitrogen digestibility coefficients, serum antioxidant activity and decline of Heterophil: Lymphocyte ratio and expression of HSP70 gene were observed in birds supplemented with 0.25 ppm Se and nano source of Se compared to inorganic Se. Significantly higher Se concentration in liver and breast muscle and higher serum Se concentration were observed in birds fed 0.25 ppm nano Se. The liver Se concentration was much higher than that of breast muscle. However, the nano Se synthesized by green method in this study did not differ significantly from the chemically synthesized nano Se. It was concluded that 0.25 ppm Se and nano form of Se are superior to lower levels and inorganic form of Se, respectively, in improving the immunity, growth, antioxidant status, and in stress alleviation of broiler chicken. However, GNS is equally efficient as chemically synthesized MNS.     

Identifying the functional groups and the influence of synthetic chelators on Cd availability and microbial biomass carbon in Cd-contaminated soil

Synthetic chelators play an important role in boosting the microbial biomass carbon (MBC), dissolved organic carbon (DOC), and heavy metal solubility in a contaminated soil toward a sustainability of environment for agricultural crops. Castor plant was grown under different levels of Cd contaminated soil (?Cd and +Cd) following adding three chelating agents, ethylenediaminetetraacetic acid (H₄EDTA), nitriloacetic acid (H₃ NTA), and NH₄ citrate (ammonium citrate) to the soil at rates of 10, 15, and 25 mmol in 5 kg of soil per pot. The highest bioavailable Cd concentrations in soil and castor plant were obtained from NH₄ citrate and H₄EDTA treatments in the contaminated soil. Fourier transform infrared (FTIR) analysis showed that NH₄ citrate was the most effective chelator in Cd-contaminated soil. MBC and DOC contents were significantly increased and reached at 81.98–80.37 and 1.96–1.90 mg kg^?1 respectively, in the (H₃ NTA) and NH₄ citrate treatments in Cd-contaminated soil. Further research is needed to investigate the use of chelators in the phytoextraction of Cd-contaminated soils under field conditions and whether it may be beneficial in accelerating the phytoextraction of Cd through hyperaccumulating plants.     

Enhanced accumulation of Cd in castor (Ricinus communis L) by soil-applied chelators

Phytoextraction has been identified as one of the most propitious methods of phytoremediation. This pot experiment were treated with varying amounts of (ethylenediamine triacetic acid) EDTA 3–15, (Nitriloacetic acid) NTA 3–10, (Ammonium citrate) NH₄ citrate 10 – 25 mmol and one mg kg^–1Cd, filled with 5 kg soil. The addition of chelators significantly increased Cd concentration in soil and plant. The results showed that maximum Cd uptake was noted under root, shoot and leaf of castor plant tissue (2.26, 1.54, and 0.72 mg kg^–1) under EDTA 15, NTA 10, and NH₄ citrate 25 mmol treatments respectively, and in soil 1.08, 1.06 and 0.52 mg kg^–1 pot^–1 under NH₄ citrate 25, NTA 10 and EDTA 15 mmol treatments respectively, as against to control (p < 0.05). Additions of chelators reduction biomass under the EDTA 15 mmol as compared to other treatments, However, Bioconcentration factor (BCF), translocation factor (TF) and remediation factor (RF) were significantly increased under EDTA 15 and NH₄ citrate 25 mmol as against control. Our results demonstrated that castor plant proved satisfactory for phytoextraction on contaminated soil, and EDTA 15 and NH₄ citrate 25 mmol had the affirmative effect on the Cd uptake in the artificial Cd-contaminated soil.     

Health risk assessments of lithium titanate nanoparticles in rat liver cell model for its safe applications in nanopharmacology and nanomedicine

Due to their high chemical stability, lithium titanate (Li₂TiO₃) nanoparticles (LTT NPs) now are projected to be transferred into different nanotechnology areas like nano pharmacology and nano medicine. With the increased applications of LTT NPs for numerous purposes, the concerns about their potential human toxicity effects and their environmental impact are also increased. However, toxicity data for LTT NPs related to human health are very limited. Therefore we aimed to investigate toxicity potentials of various concentrations (0–1,000 ppm) of LTT NPs (<100 nm) in cultured primary rat hepatocytes. Cell viability was detected by [3-(4,5-dimethyl-thiazol-2-yl) 2,5-diphenyltetrazolium bromide] (MTT) assay and lactate dehydrogenase (LDH) release, while total antioxidant capacity (TAC) and total oxidative stress (TOS) levels were determined to evaluate the oxidative injury. DNA damage was analyzed by scoring liver micronuclei rates and by determining 8-oxo-2-deoxyguanosine (8-OH-dG) levels. The results of MTT and LDH assays showed that higher concentrations of dispersed LTT NPs (500 and 1,000 ppm) decreased cell viability. Also, LTT NPs increased TOS (300, 500 and 1,000 ppm) levels and decreased TAC (300, 500 and 1,000 ppm) levels in cultured hepatocytes. The results of genotoxicity tests revealed that LTT NPs did not cause significant increases of micronucleated hepatocytes and 8-OH-dG as compared to control culture. In conclusion, the obtained results showed for the first time that LTT NPs had dose dependent effects on oxidative damage and cytotoxicity but not genotoxicity in cultured primary rat hepatocytes for the first time.     

The potential mitigation effect of ZnO nanoparticles on [Abelmoschus esculentus L. Moench] metabolism under salt stress conditions

Salt stress is known to be momentous abiotic stress which treats agricultural lands and crop production throughout the world and effects the system of food security. The current study aims to investigate the effect of foliar application of 10 mg/l of zinc oxide (ZnO) as a bulk or as a green synthesized nanoparticle (ZnO-NPs) which were hexagonal and spherical in shape and at size 16–35 nm to alleviate the impact of salinity concentrations (0, 10, 25, 50, 75 and 100% SW) on Okra (Abelmoschus esculentus L. Moench cv. Hasawi) species. The results demonstrated a gradual decrease in the photosynthetic pigments (i.e., chlorophyll a and b with total chlorophylls and carotenoids) with the growth of salinity conc. However, the sea water levels between 0 and 75% will led to increase in proline, total soluble sugar and activity of the antioxidant enzymes i.e., superoxide dismutase (SOD) and catalase (CAT) and then decrease at 100% SW. The addition of bulk ZnO or ZnO-NPs enhances the contents of the photosynthetic pigments, activity of both SOD and CAT and then lowers the accumulation of proline and total soluble sugar when equated with controls. Plants treated with ZnO-NPs showed the greatest results when compared with other treatments. The results of current study showed ZnO-NPs as an appropriate eco-friendly and low-cost application for plant growth under salinity which has an ability to moderate the salt stress effect of plants.     

Impact of chelator-induced phytoextraction of cadmium on yield and ionic uptake of maize

Enhanced phytoextraction uses soil chelators to increase the bioavailability of heavy metals. This study tested the effectiveness of ethylenediaminetetraacetic acid (EDTA) and citric acid in enhancing cadmium (Cd) phytoextraction and their effects on the growth, yield, and ionic uptake of maize (Zea mays). Maize seeds of two cultivars were sown in pots treated with 15 (Cd₁₅) or 30 mg Cd kg^?1 soil (Cd₃₀). EDTA and citric acid at 0.5 g kg^?1 each were applied 2 weeks after germination. Results demonstrated that the growth, yield per plant, and total grain weight were reduced by exposure to Cd. EDTA increased the uptake of Cd in shoots, roots, and grains of both maize varieties. Citric acid did not enhance the uptake of Cd, rather it ameliorated the toxicity of Cd, as shown by increased shoot and root length and biomass. Cadmium toxicity reduced the number of grains, rather than the grain size. The maize cultivar Sahiwal-2002 extracted 1.6% and 3.6% of Cd from soil in both Cd+ EDTA treatments. Hence, our study implies that maize can be used to successfully phytoremediate Cd from soil using EDTA, without reducing plant biomass or yield.     

Influence of EDTA on lead transportation and accumulation by Sedum alfredii hance

Hydroponics and pot experiments were conducted to study the effects of ethylenediaminetetraacetic acid (EDTA) on Pb transportation and accumulation by two contrasting ecotypes of Sedum alfredii Hance. In hydroponics experiments, the accumulating ecotype (AE) showed more ability to tolerate Pb toxicity compared with the non-accumulating ecotype (NAE). When treated with equimolar mixtures of EDTA and Pb, maximum Pb accumulation occurred without any phytotoxicity symptoms. Pot experiments with Pb contents of 400 mg kg-1 showed that 5 mM EDTA is the optimum dose for the phytoextraction of soils contaminated with relatively low Pb levels; in contrast, increasing EDTA addition resulted in increased Pb accumulation in the shoots of AE in soils with high Pb content (1200 mg kg(-1)). The post-harvest effects of EDTA on available Pb were strong compared with those without addition of EDTA (CK). Within the initial 7 days almost no differences of water-soluble Pb were noted in soils contaminated with both levels of Pb but after 2 weeks, water-soluble Pb started to decrease significantly compared with before. Considering the toxicity and biodegradability of synthetic chelators, it can be concluded that the chelate-assisted technique is more suitable for soils contaminated with low Pb levels and to avoid environment risks; a suitable dose of chelators must be considered before application.     

On the toxic effects of tetraethyl lead and its derivatives on the chrysophyte poterioochromonas malhamensis. IV. influence of lead antidotes and related agents

The unicellular alga Poterioochromonas malhamensis was exposed to 12.5 μM of inorganic or triethyl lead and simultaneously treated with lead antidotes and related agents at concentrations of 12.5, 31.25 and 62.5 μM. With increasing concentrations some of the antidotes alone slightly to severely inhibited algal growth (BAL¹, CaNa₂EDTA, EDTA, Na₂EDTA), whereas others (DPA, EGTA, DIZO) were non-toxic at the concentrations tested. EGTA and CaNa₂EDTA, at all concentrations tested, completely suppressed the growth inhibition caused by inorganic lead; Na₂EDTA and EDTA were protective at the lower or medium concentrations, but DIZO. DPA and BAL considerably enhanced lead toxicity with increasing concentrations. None of the tested agents was able to reduce the toxic effects of triethyl lead. All antidotes markedly increased inhibition of algal growth caused by triethyl lead and some were even lethal to the poisoned algae either at the highest (Na₂EDTA, EDTA, DPA) or at all concentrations used (DIZO, BAL). P. malhamensis proved to be a highly sensitive and valuable tests system and the results obtained exhibited striking parallels to medical and clinical experience in therapy of human poisoning with inorganic and organic lead compounds.     

Antioxidant defense in a lead accumulating plant, Sesbania drummondii.

Seedlings of Sesbania drummondii were grown in 500 mg l-1 Pb(NO3)2 in presence and absence of chelators: EDTA, DTPA and HEDTA for 4 weeks. Plants were assayed for activities of the antioxidant enzymes: ascorbate peroxidase (APX), guaiacol peroxidase (GPX), catalase (CAT), superoxide dismutase (SOD) and glutathione (gamma-glutamyl-cysteinyl-glycine) content. Activities of antioxidant enzymes were elevated in the presence of Pb but were similar to controls in plants grown in the presence of Pb and EDTA, -DTPA or -HEDTA. Glutathione content was significantly elevated upon exposure to Pb, but lowered upon exposure to chelators. Chlorophyll a fluorescence kinetics were assessed by determination of Fv/Fm and Fv/Fo values. Seedling growth in Pb alone and Pb + chelators did not significantly affect photosynthetic integrity (Fv/Fo) and efficiency (Fv/Fm). The results suggest that Sesbania plants were able to tolerate Pb-induced stress using an effective antioxidant defense mechanism. This study also indicates a protective role of synthetic chelators in Pb-induced oxidative stress metabolism in a Pb-accumulating plant.     

柠檬酸和EDTA对铜污染土壤环境中吊兰生长的影响

通过盆栽试验研究了在铜污染条件下,柠檬酸和EDTA作为活化剂对铜污染土壤中吊兰生长状况的影响.结果表明,柠檬酸和EDTA对吊兰富集量的影响与其对土壤中铜的活化能力呈显著性正相关.柠檬酸对土壤铜有较强的活化作用,能够有效提高吊兰对铜的吸收,且在浓度为5mmol/L时效果最为明显,而较高的铜富集量又抑制了吊兰的生长;EDTA对吊兰富集能力的影响相对较弱,对吊兰的生长也无显著影响.相比而言,柠檬酸对铜污染土壤中吊兰生长状况的影响比EDTA大.     

Effect of ZnO and TiO2 nanoparticles preilluminated with UVA and UVB light on Escherichia coli and Bacillus subtilis

We evaluated the effects of zinc oxide (ZnO) and titanium dioxide (TiO₂) nanoparticles (NPs) preilluminated with ultraviolet light on Escherichia coli and Bacillus subtilis. The experiments were conducted using three different types of light: visible, Ultraviolet A (UVA, 315–400 nm), and Ultraviolet B (UVB, 280–315 nm). The bacteria were exposed to NPs, either as liquid suspensions for growth inhibition assays or on agar plates for colony forming unit (CFU) assays. We found that the ZnO NPs were more toxic when preilluminated with UVA or UVB light than with visible light in both growth inhibition and CFU assays. TiO₂ NPs were not toxic to the bacteria under UVA or UVB preillumination conditions. The photo-dissolution of ZnO NPs increased with UV preillumination, which could explain the observed toxicity of ZnO NPs. We detected oxidative stress elicited by photoactive nanoparticles by measuring superoxide dismutase activity. The results of this study show that the toxicity of photoactive nanoparticles can be increased by UV preillumination by dissolution of toxic ions, which suggests the potential for preillumination-dependent toxicity of nanoparticles on soil environments in low light or darkness.     

不同形态氮素施肥对小黑杨幼苗生长的影响

以小黑杨当年播种苗为材料,研究了不同施氮量（12,24和48 mg·株^-1）和不同形态氮素（有机氮和无机氮）施肥对小黑杨幼苗生长的影响,以探讨小黑杨对氮基酸类有机氮素施肥的生长响应。结果表明：无论施无机氮（硝酸铵）还是有机氮（精氨酸）,小黑杨幼苗的苗高、地径、总生物量都是随施氮量的增加而增加,但是中等施氮量处理的幼苗氮利用效率最高。无机氮（硝酸铵）和有机氮（精氨酸）处理的幼苗在相同施氮水平下生长表现无明显差异,施用有机氮可以与施用无机氮获得相同的促进苗木生长的效果。不同氨基酸及其组合肥料施用对小黑杨幼苗生长的影响显著。单一氨基酸施肥情况下,施用精氨酸促进苗木生长的效果最好,谷氨酸次之,甘氨酸最差;氨基酸组合施肥情况下,有精氨酸的组合施肥苗木生长好,有甘氨酸的组合施肥苗木生长差。不同氨基酸施肥处理对小黑杨幼苗各器官氮含量没有明显影响。     

The effect of EDTA and EDDS on lead uptake and localization in hydroponically grown Pisum sativum L.

Pisum sativum plants were treated for 3 days with an aqueous solution of 100 μM Pb(NO₃)₂ or with a mixture of lead nitrate and ethylenediaminetetraacetic acid (EDTA) or [S,S]-ethylenediaminedisuccinic acid (EDDS) at equimolar concentrations. Lead decline from the incubation media and its accumulation and localization at the morphological and ultrastructural levels as well as plant growth parameters (root growth, root and shoot dry weight) were estimated after 1 and 3 days of treatment. The tested chelators, especially EDTA, significantly diminished Pb uptake by plants as compared to the lead nitrate-treated material. Simultaneously, EDTA significantly enhanced Pb translocation from roots to shoots. In the presence of both chelates, plant growth parameters remained considerably higher than in the case of uncomplexed Pb. Considerable differences between the tested chelators were visible in Pb localization both at the morphological and ultrastructural level. In Pb+EDTA-treated roots, lead was mainly located in the apical parts, while in Pb+EDDS-exposed material Pb was evenly distributed along the whole root length. Transmission electron microscopy and EDS analysis revealed that in meristematic cells of the roots incubated in Pb+EDTA, large electron-dense lead deposits were located in vacuoles and small granules were rarely noticed in cell walls or cytoplasm, while after Pb+EDDS treatment metal deposits were restricted to the border between plasmalemma and cell wall. Such results imply different ways of transport of those complexed Pb forms.     

纳米氧化锌和接种丛枝菌根真菌对大豆生长及营养吸收的影响

纳米氧化锌是应用最广的人工纳米颗粒(nanoparticles,NPs)之一,具有一定生物毒性。丛枝菌根(arbuscular mycorrhizal,AM)真菌能与陆地上80%以上的高等植物形成丛枝菌根共生体,并能改善宿主植物矿质营养,提高其抗逆性。然而纳米ZnO与丛枝菌根的关系尚不清楚。通过温室沙培盆栽试验,研究了施加不同水平纳米ZnO(0、500、1000、2000、3000 mg/kg)和接种AM真菌Acaulospora mellea对大豆生长及营养状况的影响。结果表明,3000 mg/kg的纳米ZnO显著抑制大豆植株生长,表现出植物毒性,在其他水平时没有显著影响。纳米ZnO在施加水平500、1000 mg/kg时没有抑制AM真菌对大豆根系的侵染,但是高施加水平(2000 mg/kg)时对AM真菌产生毒害,几乎完全抑制大豆根系菌根侵染。接种AM真菌仅在500 mg/kg纳米ZnO时显著促进大豆生长,增加大豆植株对P、K、N的吸收,降低根系Zn含量。纳米ZnO可能会持续释放锌离子,并抑制大豆根系对矿质营养元素的吸收,从而产生生物毒性,而AM真菌与大豆根系的共生可起到有益作用。     

Effects of low pH and aluminum stresses on common beans (Phaseolus vulgaris) differing in low-phosphorus and photoperiod responses

Using common beans differing greatly in the response to photoperiod and low-phosphorus (P) stress, we investigated their responses to acidity and aluminum (Al)toxicity and the relationship between Al tolerance and organic acid exudation under Al or low P stress. A genotype Ginshi was found to be sensitive to low pH treatment. When exposed to pH 4.5, serious curvature in the root tips of cv. Ginshi was observed; however, it was completely corrected by the application of 5 or 10 μmol/L AlCl3; increasing calcium (Ca) could ameliorate Al toxicity, but it could not correct root curvature at pH 4.5. Common beans showed significant differences in both root growth and Al tolerance, and the varieties from the Andes were more tolerant to Al toxicity than those from the Mesoamerican origin. In the presence of 50 μmol/L AlCl3,all the common bean genotypes exuded citrate, and a significant difference in the amounts of citrate was observed among genotypes. The genotypes originated in the Mesoamerica tended to release more citrate than other origins in the presence of Al. The P-inefficient genotype DOR364 exuded more citrate than the P-efficient genotype G19833 in the presence of 50 μmol/L AlCl3, whereas no organic acids were detected in root exudates under low-P stress. A reduction of citrate exudation in the DOR364, but a slight increase of citrate exudation in the G19833, was observed under Al stress after they were exposed to 6-d P starvation. These results suggest that different low-P or Al tolerance in common beans might not be associated with organic acid exudation.     

Impact of Bulk and Nanosized Titanium Dioxide (TiO<Subscript>2</Subscript>) on Wheat Seed Germination and Seedling Growth

The impacts of different concentrations of bulk and nanosized TiO₂ on seed germination and seedling growth of wheat were studied in a randomized completely design with four replications in the College of Agriculture, Ferdowsi University of Mashhad, Iran, in 2011. The experimental treatments included five concentrations of bulk (1, 2, 10, 100, and 500 ppm), five concentrations of nanosized TiO₂ (1, 2, 10, 100, and 500 ppm), and control (without any TiO₂). Results indicated that among the wheat germination indices, only mean germination time was affected by treatments. The lowest and the highest mean germination time (0.89 vs. 1.35 days) were obtained in 10 ppm concentration of nanosized TiO₂ and control treatments, respectively. In addition, shoot length, seedling length, and root dry matters were affected by bulk and nanosized TiO₂ concentrations, significantly. Shoot and seedling lengths at 2 and 10 ppm concentrations of nanosized TiO₂ were higher than those of the untreated control and bulk TiO₂ at 2 and 10 ppm concentrations. Employing nanosized TiO₂ in suitable concentration could promote the seed germination of wheat in comparison to bulk TiO₂ but in high concentrations had inhibitory or any effect on wheat.     

Mobilisation of recently absorbed Fe in ex vivo perfused rat duodena and the influence of iron status and subsequently absorbed chelators

To investigate the effect of subsequently absorbed metal chelators on recently absorbed ⁵⁹Fe, duodenal segments from iron-deficient and iron-adequate rats were perfused ex vivo until the ⁵⁹Fe tissue load had reached a steady state. Subsequently, the segments were perfused with 3 model chelators and their iron complexes: nitrilotriacetic acid (NTA), ethylenediaminetetraacetic acid (EDTA) and citrate. Of these, NTA and EDTA bind iron much tighter than citrate, and Fe–NTA complexes exchange iron within seconds while Fe-EDTA complexes need 48 h to reach equilibrium.

Duodenal mucosa-to-serosa transport rates were comparable for all 3 chelators and correlated linearly with luminal concentration. Subsequent perfusion with increasing NTA, Fe–NTA(1:2) and EDTA concentrations mobilised increasing amounts of ⁵⁹Fe from the duodenum. Mobilised ⁵⁹Fe moved preferentially back into the luminal perfusate in iron-adequate segments. In iron-deficient segments, ⁵⁹Fe preferentially continued the absorption process across the basolateral membrane. Fe–EDTA(1:1) hardly mobilised any ⁵⁹Fe back into the lumen, though basolateral transfer increased at high concentrations. Citrate and Fe–citrate(1:1) mobilised ⁵⁹Fe only at very high concentrations.

This behaviour is in accordance with the rules of complex chemistry: strong, fast reacting ligands like NTA show most impact. Slowly reacting complexes like Fe–EDTA(1:1) have little mobilising impact in spite of strong affinity between EDTA and iron. The low affinity between iron and citrate can be compensated by large concentration. Moreover, iron-deficient segments show stronger re-uptake of mobilised ⁵⁹Fe from the lumen and a stronger transfer of ⁵⁹Fe from the tissue across the basolateral membrane. Both are compatible with the more marked expression of divalent metal transporter 1 (DMT-1) and IREG-1 at the brushborder and basolateral membrane of iron-deficient enterocytes. The data suggest that iron ions interact with food ligands during their passage from the apical to the basolateral side of duodenal enterocytes.     

Engineered ZnO and TiO(2) nanoparticles induce oxidative stress and DNA damage leading to reduced viability of Escherichia coli

Extensive use of engineered nanoparticle (ENP)-based consumer products and their release into the environment have raised a global concern pertaining to their adverse effects on human and environmental health. The safe production and use of ENPs requires improvement in our understanding of environmental impact and possible ecotoxicity. This study explores the toxicity mechanism of ZnO and TiO(2) ENPs in a gram-negative bacterium, Escherichia coli. Internalization and uniform distribution of characterized bare ENPs in the nano range without agglomeration was observed in E. coli by electron microscopy and flow cytometry. Our data showed a statistically significant concentration-dependent decrease in E. coli cell viability by both conventional plate count method and flow cytometric live-dead discrimination assay. Significant (p<0.05) DNA damage in E. coli cells was also observed after ENP treatment. Glutathione depletion with a concomitant increase in hydroperoxide ions, malondialdehyde levels, reactive oxygen species, and lactate dehydrogenase activity demonstrates that ZnO and TiO(2) ENPs induce oxidative stress leading to genotoxicity and cytotoxicity in E. coli. Our study substantiates the need for reassessment of the safety/toxicity of metal oxide ENPs.     

Secretion of citrate from roots in response to aluminum and low phosphorus stresses in Stylosanthes

Excess aluminum (Al) ions and phosphorus (P) deficiency are the key factors that limit plant growth in acid soils. Secretion of organic acids (OA) from roots has been proposed as an Al-resistance mechanism. Nonetheless, the correlation between Al resistance and this mechanism has not been tested beyond a very small number of Al-resistant and Al-sensitive genotypes. To elucidate the mechanisms responsible for plant adaptability to acid soils, we studied the secretion of OA from roots of Stylosanthes in response to high-Al and low-P stresses using six different genotypes. Relative root inhibition by 50?µM Al ranged from 25–71% and differed significantly among six Stylosanthes genotypes. Al treatment induced the secretion of citrate from the roots of Stylosanthes seedling in a dose- and time-dependent manner. Moreover, the secretion rate was significantly higher in the Al-resistant genotype. On the other hand, inhibition of Al-induced citrate secretion by phenylisothiocyanate or 9-anthracenecarboxylic acid resulted in an increase in Al content in Stylosanthes root apices. P deficiency also induced citrate secretion from Stylosanthes seedling roots. Furthermore, citrate secretion was much more robust with exposure to both excess-Al and P-deficiency stresses than under either stress alone. Unlike Al-induced citrate secretion, which was rapid, low-P-induced secretion was a slow process, with significant increases in secretion only becoming evident after 6 d of treatment with free phosphate. The lag between treatment with Al and citrate secretion was approximately 4 h. These results suggest that the secretion of citrate is a mechanism for resistance to both excess-Al and low-P stresses in Stylosanthes.