Lucas Anjos Souza

Lead tolerance and phytoremediation potential of Brazilian leguminous tree species at the seedling stage.

A greenhouse pot experiment was conducted to evaluate the potential of three Brazilian leguminous woody species, Mimosa caesalpiniaefolia, Erythrina speciosa and Schizolobium parahyba, for the revegetation of lead- (Pb-) contaminated areas. The response of seedlings to increasing Pb concentrations (0, 250, 500 and 1000 mg kg(-1)) in the soil was studied. In addition to Pb accumulation and translocation, the following parameters were assessed: chlorophyll, nitrate, ammonia, lipid peroxidation (MDA) and free amino acid content; seedling growth; and nitrogenase activity. No differences were observed in the germination of woody species seeds sown in soils with or without Pb addition. M. caesalpiniaefolia did not show visual symptoms of Pb toxicity, while the other two species demonstrated stress symptoms, including reduced shoot biomass yield, leaf area and height. Biochemical analyses of plant tissues revealed markedly different responses to increasing Pb concentrations, such as changes in foliar soluble amino acid composition in S. parahyba; changes in ammonia and nitrate content in E. speciosa, M. caesalpiniaefolia and S. parahyba; and changes in MDA content in S. parahyba. The levels of chlorophyll a and b and carotenoid were affected in the species studied. For the Nitrogen-fixing (N(2)-fixing) species E. speciosa, an increase of Pb in the soil affected nodule formation and growth, which led to reduced nitrogenase activity in seedlings. The concentration of Pb in shoots and roots increased with the Pb concentration in soil. However, most of the Pb absorbed accumulated in the roots, and only a small fraction was translocated to aboveground parts. These findings were confirmed by the low bioconcentration factor (BCF) and translocation factor (TF) values for the three species. The tolerance index (TI) values suggested that M. caesalpiniaefolia, a N(2)-fixing tree, was the species that was most tolerant to high Pb concentrations in soil, while E. speciosa and S. parahyba showed moderate tolerance. Of the three Brazilian native woody species studied, M. caesalpiniaefolia was found to have the highest Pb tolerance and phytostabilisation potential in Pb-contaminated soils.

Use of non-hyperaccumulator plant species for the phytoextraction of heavy metals using chelating agents

Soil contamination by heavy metals is a challenge faced by many countries, and engineering technologies to solve this problem are expensive and can cause negative impacts on the environment. One way to minimise the levels of heavy metals in the soil is to use plants that can absorb and accumulate heavy metals into harvestable parts, a process called phytoextraction. Typical plant species used in research involving phytoextraction are heavy metal hyperaccumulators, but plants from this group are not good biomass producers and grow more slowly than most species; thus, they have an important role in helping scientists understand the mechanisms involved in accumulating high amounts of heavy metals without developing symptoms or dying. However, because of their slow growth, it is not practical to use these species for phytoextraction. An alternative approach is to use non-hyperaccumulator plants assisted by chelating agents, which may improve the ability of plants to accumulate more heavy metals than they would naturally. Chelating agents can be synthetic or organic acids, and the advantages and disadvantages of their use in improving the phytoextraction potential of non-hyperaccumulator plants are discussed in this article. We hope to draw attention to ways to improve the phytoextraction potential of non-hyperaccumulator plants that produce a large amount of biomass and to stimulate more research on phytoextraction-inducing substances.

Evaluation of Mycorrhizal Influence on the Development and Phytoremediation Potential of Canavalia Gladiata in Pb-Contaminated Soils

Soil contamination by heavy metals is a serious problem to humans due to its high level of toxicity. The heavy metal lead (Pb) is commonly used in industries and if the disposal of residues that contain this element is not done properly may result in tragic consequences to the organisms. In this experiment we assessed the potential of a forrage leguminous, Canavalia gladiata, to phytoremediate lead-contaminated soil under mycorrhizal influence. The experimental design was composed of 4 Pb doses (0, 250, 500, and 1000 mg kg−1 of soil) and the plants were inoculated or uninoculated with arbuscular mycorrhizal fungi (AMF). We observed that the nodulation was severely affected by the presence of Pb independently of the mycorrhizal status; most of the elements analyzed were affected independently of the mycorrhizal status with exception of P. The mycorrhizal colonization was able to restrict the entrance of Pb in plants under high concentrations of Pb but promoted its accumulation in both organs under intermediate concentrations of this element. Besides the mycorrhization did not promote plant growth under Pb stress, the use of this plant may be considered to be used for phytostabilization purposes.

Mycorrhization alters foliar soluble amino acid composition and influences tolerance to Pb in Calopogonium mucunoides

Soil contamination by lead (Pb) is a problem due to the persistence of this element on soil. High amounts of Pb in soil impair plant establishment, however some plants may increase tolerance to heavy metals when colonized by arbuscular mycorrhizal fungi (AMF). The leguminous plant, Calopogonium mucunoides, is Pb-sensitive that is tolerant to Pb when associated to AMF. We performed a chromatographic analysis of foliar free-amino acid in non-mycorrhizal and mycorrhizal plants to determine its relation with Pb tolerance. Mycorrhization caused drastic increase in aspartate, glutamine, glycine, threonine, alanine, isoleucine and gamma-aminobutiric acid (GABA), while depletion of asparagine, histidine and arginine was observed in AMF-associated plants. When grouped according to common metabolic precursor, it was found that amino acids derived from 3-phosphoglicerate and pyruvate was higher in mycorrhizal plants while amino acids derived from glucose-6-phosphate and 2-oxoglutarate were higher in non-mycorrhizal plants; phosphoenolpyruvate and oxaloacetate pathways were not influenced by mycorrhization. Summarizing, mycorrhization changed soluble amino acids profile in C. mucunoides leaves, especially aspartate, alanine and GABA, which may be involved in tolerance to abiotic stress. Additionally the depletion of asparagine, histidine and arginine may be related to a deviation for metabolic pathways not related to protein biosynthesis but to the synthesis of polyamines, especially in the case of arginine. Therefore, we suggest that mycorrhization influence on soluble free amino acid profile in leaves can be one of the factors involved with the attenuation of Pb toxicity in AMF-associated C. mucunoides plants.

Tolerância e potencial fitorremediador de Stizolobium aterrimum associada ao fungo micorrízico arbuscular Glomus etunicatum em solo contaminado por chumbo

Heavy metal pollution of soils has increased significantly in the last years owing to anthropic action. Several techniques can be used to revert or to minimize soil contamination, although many of these techniques are harmful to the soil. An alternative is to use a new technique, called phytoremediation, based on the ability of plants to take up elements from soils with excessive high levels of metals or of other potentially toxic elements and thus contribute to soil decontamination. The inoculation of plants with arbuscular mycorrhizal fungi (AMF) can influence the absorption of these elements. The phytoremediation potential of Stizolobium aterrimum plants with or without AMF in soils with increasing lead concentrations was evaluated in a greenhouse experiment, in a 4 x 2 factorial design. The treatments consisted of the addition of four Pb rates (0, 250, 500 and 1000 mg dm-3) to the soil where black velvet bean plants associated or not with Glomus etunicatum AMF where grown. The results showed that black velvet bean was Pb tolerant at the tested rates. The association with AMF did not influence the Pb plant uptake. However, the mycorrhiza influenced biological nitrogen fixation by increasing the activity of the enzyme nitrogenase in mycorrhizal plants. Despite the good results obtained in relation to Pb tolerance of black velvet bean, further studies on the uptake of this element are needed, above all in the case of multicontaminated soils, as actually observed in polluted systems.

Physiological and biochemical responses of Dolichos lablab L. to cadmium support its potential as a cadmium phytoremediator

PurposeThis study aims to investigate the response of a high biomass producer non-hyperaccumulator legume plant species, Dolichos lablab L., to cadmium (Cd) stress for phytoremediation process.Materials and methodsThree individual experiments were carried out to assess physiological and biochemical parameters to support the use of this plant species as a phytoremediator. The first experiment was carried out in Cd-contaminated soil while the second and third experiments were conducted in sand in which Cd was applied to study biochemical responses. Analysis of mineral nutrition, phytoremediation parameters, antioxidant response, and protein identification by gel-based proteomics were performed.Results and discussionGood tolerance to Cd under moderate level of contamination was observed. Mineral nutrition was little affected, and phytoremediation index was satisfactory. Additionally, biochemical responses based on antioxidant enzyme analysis were well responsive in roots, reflecting the capacity of Cd stress attenuation in this organ. A proteomic analysis revealed positive regulation of root proteins involved in carbohydrate, amino acids, nitrogen metabolism, and abiotic/biotic stress response, which together may contribute to create a scenario to overcome Cd-induced stress.ConclusionsBased on the physiological and biochemical results, we concluded that D. lablab L. is suitable for phytoremediation/phytostabilization purposes.

pH effects on nodulation and biological nitrogen fixation in Calopogonium mucunoides

Calopogonium mucunoides Desv. is able to form nodules, root organs in which biological nitrogen fixation takes place, after a symbiotic interaction with soil bacteria known as rhizobia. Such distinct advantage of some legume species faces different environmental abiotic factors such as acid and alkaline pH conditions of soil. Nodulation and symbiotic nitrogen fixation response in different pH ranges were determined under greenhouse conditions. Plants were cultivated in vermiculite and treated with nutritive solution adjusted to pH 4.0, 5.5, and 7.0, and after three months, the following variables were measured: nodule number, nitrogen fixation, tissue protein, amino acids, total ureides, allantoin, and allantoate. The number of nodules and nitrogen fixation were enhanced under acidic conditions, but nitrogenase activity was drastically decreased at pH 7.0. Acidic conditions decreased the amount of protein, amino acids, total ureides, allantoate, and allantoin in leaves and nodules, but at pH 5.5, only protein content was decreased. Symbiosis with C. mucunoides and biological nitrogen fixation were kept under acidic conditions, but it was negatively affected under conditions that are near to neutral. It is reasonable to conclude that the association between C. mucunoides and nitrogen-fixing bacteria from “Cerrado” evolved in this context, which is supported by the difficulties faced to keep interaction under pH near neutrality.

Changes in soluble amino acid composition during Canavalia ensiformis development: responses to nitrogen deficiency

A limited number of studies focusing nitrogen metabolism have been carried out with plants from Cerrado, the second largest biome of Brazil. Canavalia ensiformes is a legume native from Cerrado and is considered an important forage crop that contributes to soil nitrogen (N) improvement. There are few studies related to amino acid metabolism and growth capacity under nitrogen deficiency for this plant species. Therefore, the objective of this work was to study the profile of soluble amino acids during the growth cycle (from vegetative to the reproductive stage) of Canavalia ensiformis. Major changes in the concentration and composition of soluble amino acids at the beginning of the reproductive stage were observed indicating important alterations in amino acids metabolism. The data revealed that N-stress conditions led to increased aspartate and decreased asparagine contents in most organs and developmental stages of C. ensiformis. A pronounced increase in glutamate concentration during N-stress was also detected. Glutamine, alanine, GABA, threonine, histidine, arginine and glycine metabolisms were probably impaired by N deficiency, which was dependent upon plant developmental stage. The effect of nitrate presence or absence on amino acids metabolism in C. ensiformis is discussed.

Comprehensive in vitro and in vivo studies of novel melt-derived Nb-substituted 45S5 bioglass reveal its enhanced bioactive properties for bone healing

The present work presents and discusses the results of a comprehensive study on the bioactive properties of Nb-substituted silicate glass derived from 45S5 bioglass. In vitro and in vivo experiments were performed. We undertook three different types of in vitro analyses: (i) investigation of the kinetics of chemical reactivity and the bioactivity of Nb-substituted glass in simulated body fluid (SBF) by 31P MASNMR spectroscopy, (ii) determination of ionic leaching profiles in buffered solution by inductively coupled plasma optical emission spectrometry (ICP-OES), and (iii) assessment of the compatibility and osteogenic differentiation of human embryonic stem cells (hESCs) treated with dissolution products of different compositions of Nb-substituted glass. The results revealed that Nb-substituted glass is not toxic to hESCs. Moreover, adding up to 1.3 mol% of Nb2O5 to 45S5 bioglass significantly enhanced its osteogenic capacity. For the in vivo experiments, trial glass rods were implanted into circular defects in rat tibia in order to evaluate their biocompatibility and bioactivity. Results showed all Nb-containing glass was biocompatible and that the addition of 1.3 mol% of Nb2O5, replacing phosphorous, increases the osteostimulation of bioglass. Therefore, these results support the assertion that Nb-substituted glass is suitable for biomedical applications.

Boron phytoremediation: Stizolobium aterrimum is tolerant and can be used for phytomanagement of boron excess in soils

ABSTRACT Soil contamination by toxic elements causes concern and is increasing through industrial development, mining activities and the overuse of chemical fertilisers. Some toxic elements, such as boron, also play a structural role. The present study aimed to evaluate the behaviour of S. aterrimum plants growing in boron artificially contaminated soils. It has been found that S. aterrimum is tolerant to low doses of boron, moderately tolerating doses of up to 240 mg.dm−3. But high doses (doses above the adequate zone) cause decreased plant growth and present symptoms of toxicity, such as foliar chlorosis. The dose of 240 mg.dm−3 can already be considered toxic, since the tolerance index (TI) was less than 50%. The species can be used as a phytoextractor in low doses of boron because it presented a high tolerance index (TI) and demonstrated the ability to uptake and accumulate boron in root tissue.