Isolation and characterization of microsatellite loci in Astroniumgraveolens (Anacardiaceae) and cross amplification in related species.

Microsatellite markers are a useful genetic tool to answer ecological and conservation genetics questions. Microsatellite primers were developed and characterized to evaluate forest fragmentation effects on genetic structure, diversity and gene flow patterns in the dioecious tropical tree Astroniumgraveolens (Anacardiaceae). Using genomic library enrichment, sixteen microsatellite loci were developed for A.graveolens. Polymorphism was evaluated in 80 individuals from four localities in Mexico. The overall number of alleles detected was 63, average alleles per locus 3.9 with a range from one to 11 per locus. Cross amplification trails on related species in the Anacardiaceae family: Spondiaspurpurea and Amphipterygiumadstringens, achieved successful amplification for all primers. Microsatellite markers described here are the first to be characterized for A.graveolens. These genetic markers will be a useful tool to assess the genetic consequences of habitat fragmentation and selective logging on A.graveolens populations. Cross amplification success on S.purpureaandA.adstringens suggest that they may be used for population genetic studies in other species within the family.

Tolerance evaluation and morphophysiological responses of Astronium graveolens, a native brazilian Cerrado, to addition of lead in soil.

Since the ecosystem is contaminated by lead, decontamination is a difficult and expensive process. Therefore, an alternative would be the use of phytoremediation plants, which have been studied more intensely in recent decades. Astronium graveolens Jacq (Anacardiaceae) is a Cerrado native species and plants of this biome are known to present adaptations and modifications that keep them in this ecosystem. Our aim was to find the tolerance index of A. graveolens to lead doses and to evaluate the morphophysiological alterations of the species when exposed to the heavy metal. The experiment was carried out in a greenhouse in Ilha Solteira-SP, with a completely randomized design and using lead acetate (Pb(C2H3O2)2) as the source of the heavy metal. Samples of the vegetative organs (roots and leaves) were fixed, subsequently dehydrated in an ethyl series, included in hydroxyethyl methacrylate (Leica Historesin) and sectioned for histological slide assembly and subsequent anatomical analysis. The levels of phenolic compounds, protein, amino acid, ammonia, allantoic acid and allantoin were quantified. We calculated the tolerance index for the species. Significant differences were found in leaf and root tissues anatomy, while in relation to the physiology of A. graveolens, a significant difference was observed when the concentration of total ureids in the roots was evaluated. Pb did not interfere with the survivability of the species. In fact, A. graveolens showed a higher secondary growth in the treatment with higher level of lead.

Mutagenicity and chemopreventive activities of Astronium species assessed by Ames test.

In the neotropical savannah, Astronium species are used in popular medicine to treat allergies, inflammation, diarrhea and ulcers. Given that natural products are promising starting points for the discovery of novel potentially therapeutic agents, the aim of the present study was to investigate the mutagenic and antimutagenic activities of hydroalcoholic extracts of Astronium spp. The mutagenicity was determined by the Ames test on Salmonella typhimurium strains TA98, TA97a, TA100 and TA102. The antimutagenicity was tested against the direct-acting and indirect-acting mutagens. The results showed that none of the extracts induce any increase in the number of revertants, demonstrating the absence of mutagenic activity. On the other hand, the results on the antimutagenic potential showed a moderate inhibitory effect against NPD and a strong protective effect against B[a]P and AFB1. This study highlights the importance of screening species of Astronium for new medicinal compounds. The promising results obtained open up new avenues for further study and provide a better understanding the mechanisms by which these species act in protecting DNA from damage. However, further pharmacological and toxicological investigations of crude extracts of Astronium spp., as well as of its secondary metabolites, are necessary to determine the mechanism(s) of action to guarantee their safer and more effective application to human health.

Morphological responses and tolerance of a tree native to the Brazilian Cerrado Astronium fraxinifolium Schott to boron toxicity.

The indiscriminate use of fertilizers and chemical pesticides can lead to boron contamination of the soil. Decontamination in general is expensive and results in other impacts. Phytoremediation is a sustainable alternative for soil restoration. Astronium fraxinifolium Schott (Anacardiaceae) is a tree species native to the Cerrado that is considered to be a pioneer species and a selective xerophyte, and it has been widely used in the reforestation and restoration of degraded areas. This study set out to characterize the physiology and anatomy of A. fraxinifolium under different boron concentrations and to assess the tolerance and phytoremediation potential of the species. An experiment with a completely randomized design was conducted in a greenhouse. The carbon allocation and chlorophyll content of leaves of A. fraxinifolium were determined. Boron concentration and the species' tolerance index were calculated from root and shoot samples. Levels of amino acids, proteins, total carbohydrates, starch, phenolic compounds, and anatomical analysis were also measured. A. fraxinifolium showed tolerance to boron concentrations in the substrate and accumulated a greater amount of the element in the aerial part, showing its phytoextraction ability. No significant differences were found in the physiology of A. fraxinifolium; however, some anatomical changes were observed. In the leaves, there were changes in the thickness of the abaxial surface of the epidermis and palisade and spongy parenchyma, and total leaf thickness, and in the roots, there were changes in the thickness of the phloem, diameter of vessel elements, and number of vessel elements per square millimeter. However, boron did not interfere in the development and survival of A. fraxinifolium, which points to the possibility that the species has phytoremediation potential.

Antifungal Activity of a Hydroethanolic Extract From Astronium urundeuva Leaves Against Candida albicans and Candida glabrata.

We have previously reported on the activity of different extracts from Astronium sp. against Candida albicans, with the hydroethanolic extract prepared from leaves of A. urundeuva, an arboreal species widely distributed in arid environments of South America and often used in folk medicine, displaying the highest in vitro activity. Here we have further evaluated the antifungal activity of this extract against strains of C. albicans and C. glabrata, the two most common etiological agents of candidiasis. The extract was tested alone and loaded into a nanostructured lipid system (10% oil phase, 10% surfactant and 80% aqueous phase, 0.5% Poloxamer 407®). In vitro susceptibility assays demonstrated the antifungal activity of the free extract and the microemulsion against both Candida species, with increased activity against C. glabrata, including collection strains and clinical isolates displaying different levels of resistance against the most common clinically used antifungal drugs. Checkerboard results showed synergism when the free extract was combined with amphotericin B against C. albicans. Serial passage experiments confirmed development of resistance to fluconazole but not to the free extract upon prolonged exposure. Although preformed biofilms were intrinsically resistant to treatment with the extract, it was able to inhibit biofilm formation by C. albicans at concentrations comparable to those inhibiting planktonic growth. Cytotoxicity assays in different cell lines as well as an alternative model using Artemia salina L. confirmed a good safety profile of the both free and loaded extracts, and an in vivo assay demonstrated the efficacy of the free and loaded extracts when used topically in a rat model of vaginal candidiasis. Overall, these results reveal the promise of the A. urundeuva leaves extract to be further investigated and developed as an antifungal.

Synergism between ozone and light stress: structural responses of polyphenols in a woody Brazilian species.

Microscopic studies on isolated ozone (O3) effects or on those in synergy with light stress commonly report the induction of polyphenols that exhibit different aspects within the vacuole of photosynthesizing cells. It has been assumed that these different aspects are randomly spread in the symptomatic (injured) regions of the leaf blade. Interestingly, secretory ducts that constitutively produce polyphenols also exhibit these same variations in their vacuolar aspect, in a spatial sequence related to the destiny of these cells (e.g., programmed cell death (PCD) in lytic secretion processes). Here, we demonstrate that the deposition pattern of polyphenols prior to the establishment of the hypersensitive-like response, a type of PCD caused by O3, follows the same one observed in the epithelial cells of the constitutive lysigenous secretory ducts. Astronium graveolens, an early secondary Brazilian woody species, was selected based on its susceptibility to high light and presence of secretory ducts. The synergism effects were assessed by exposing plants to the high O3 concentrations at an urban site in São Paulo City. Confocal, widefield and light microscopies were used to examine polyphenols' occurrence and aspects. The spatial pattern of polyphenols distribution along the leaflets of plants submitted to the synergism condition, in which a dense vacuolar aspect is the target of a cell destined to death, was also observed in the constitutive secretory cells prior to lysis. This similar structural pattern may be a case of homology of process involving both the constitutive (secretory ducts) and the induced (photosynthesizing cells) defenses.

Responses of a tropical tree species to ozone: visible leaf injury, growth, and lipid peroxidation.

The Brazilian native tree species Astronium graveolens was indicated as sensitive to ozone in a fumigation experiment. Thus, the objective of this study was to evaluate how sensitive A. graveolens is to ozone under realistic conditions in the field. Eighteen saplings were exposed to ozone in a contaminated area and in a greenhouse with filtered air during two exposure periods of approximately 63 days each (March-May 2012 and September-October 2012). Leaf injury was analyzed by means of its incidence and severity, the leaf injury index (LII) and the progression of leaf abscission. These variables were monitored weekly, whereas growth and lipid peroxidation were monitored monthly. Plants exposed to ozone showed significant growth decrease and visible leaf injury increase, but lipid peroxidation and leaf abscission remained unchanged. These results indicated that plants subjected to ozone possibly diverted energy from growth to the production of antioxidants necessary to cope with ozone-induced oxidative stress.

Antioxidant and antiangiogenic activity of Astronium graveolens Jacq. leaves.

Angiogenesis is involved in many physiological and pathological conditions. Natural compounds with antioxidant activity have also been reported to possess potent antiangiogenic properties by regulating angiogenesis modulators such as vascular endothelial growth factor (VEGF). Based on this, we screened the antioxidant and antiangiogenic activities of Astronium graveolens leaf extracts by a DPPH test and a competitive enzyme-linked immunosorbent assay, respectively. MeOH extract expressed a significant free radical-scavenging activity (EC50 = 37.65 µg/mL) and it was able to inhibit the interaction between placental growth factor (PlGF) (placental growth factor), a VEGF family member, and its receptor Flt-1 by more than 50% at 1 mg/mL. 1,2,3,4,6-Penta-O-galloyl-d-glucopyranose, 6 is the most active compound of the extract. It exhibited a high potency in scavenging DPPH (EC50 = 2.16 µg/mL) and reduced by 58% the PlGF/Flt-1 interaction at a concentration of 50 µM. Moreover, the known compounds (1-6) have been isolated for the first time in A. graveolens.

Development and optimization of extractives from Astronium urundeuva (allemão) Engl. by factorial design

The purpose of this study was to evaluate the influences from variables of preparation on dry residue and total tannins from macerates of Astronium urundeuva barks. Thus, a 3 ² factorial design was used to study the importance of plant proportion (10, 15 and 20 percent, w/v) and the ethanol concentration (40, 60 and 80 percent, v/v) on the selected response. Mathematical models were fitted according to experimental data. The validated equations were used to generate response surfaces. The analysis of the surfaces showed that the optimum conditions to obtain extractive preparations with high extractive efficiency for tannins were: 10 percent (w/v) of plant with ethanol 40 percent (v/v) as solvent.

O objetivo deste trabalho foi avaliar as influências de variáveis de preparação de macerados das cascas de Astronium urundeuva sobre resíduo seco e teor de taninos totais. Desta forma, um desenho fatorial do tipo 3² foi empregado para avaliar a importância da proporção de planta (10, 15 e 20 por cento; m/v) e da concentração de etanol (40, 60 e 80 por cento; v/v) sobre as respostas eleitas. Modelos matemáticos foram ajustados aos dados experimentais. As equações validadas foram usadas para gerar superfícies de respostas. A análise das superfícies demonstrou que as condições ótimas para obtenção de preparações extrativas com elevada eficiência extrativa de taninos foram: 10 por cento (m/v) para planta e etanol 40 por cento (v/v) como solvente.