Vascular traits of stem galls: Cell increment versus morphogenetic constraints in wood anatomy.

Eremanthus erythropappus hosts globoid stem galls induced by Neolasioptera sp. (Diptera: Cecidomyiidae) close to the stem apex, which do not compromise the shoot apical meristem (SAM). We hypothesize that maintenance of the SAM, as well as the increasing number of leaves per branch and of galled stem lengths and diameters, are a consequence of alterations in vascular cells and, consequently, in the priority for water flow from non-galled stems to the stem galls. Our study focuses on the globoid stem galls to evaluate if gall induction and development promote changes in structure and function of secondary xylem cells. Anatomical, cytological, histometric and physiological methods were used to analyse non-galled stem branches (NGS), mature globoid stem galls and stem portions below and above the galls. These analyses revealed that vessel elements are larger in stem galls and in stem portions above the galls. Under Neolasioptera sp. induction activity, the vascular cambium of E. erythropappus produces less numerous but larger vessel elements and overproduces parenchyma cells. Contrary to the vascular constriction hypothesis proposed for bacterial galls, the vascular traits of the Neolasioptera sp. stem galls on E. erythropappus result in priority for water flow to galls and the non-galled portions above the galls, allowing the maintenance of galled stem growth and development.

Effects of fluoride emission on the morphoanatomy of three plant species endemics to Brazil using passive biomonitoring.

Fluoride is the most phytotoxic atmospheric pollutant. The objective of this study was to evaluate the effects of fluoride emissions by an aluminum smelter on three plant species endemics to Brazil, located at Parque Estadual do Itacolomi (PEI). The monitored species were Byrsonima variabilis (Malpighiaceae), Myrceugenia alpigena (Myrtaceae), and Eremanthus erythropappus (Asteraceae), which were monitored during 9 months using passive biomonitoring at five different locations with different distances from the smelter. The monitored species did not show macroscopic phytotoxicity damage to fluoride; however, they did show microscopic damage. The species closer to the smelter presented more severe anatomic damages, such as rupture of cell walls, protoplast retraction, and trichome alterations. Damaged stomatal ledges, flaking epicuticular wax, and damages to trichomes were observed. M. alpigena showed a higher accumulation of fluoride than the other species at all monitored sites. The test for cell death with Evans Blue was positive for the three studied species. Through biomonitoring in the PEI, we concluded that the emissions from the aluminum smelter affect the native vegetation and that due to the greater accumulation of fluoride and the diversity of microscopic damage in M. alpigena, the use of this species in the monitoring of environments polluted by fluoride is enhanced.

Antileishmanial activity of the essential oils of Myrcia ovata Cambess. and Eremanthus erythropappus (DC) McLeisch leads to parasite mitochondrial damage.

Leishmania amazonensis is a species causative of cutaneous and anergic diffuse cutaneous leishmaniasis, treatment-resistant form, in the New World. Plants essential oils exhibit great potential as microbicide agents. We described the composition of the essential oils of two plants native from Brazil, Myrcia ovata, with geranial and neral as major constituents, and Eremanthus erythropappus, with α-bisabolol. In vitro effects of these essential oils on L. amazonensis promastigotes growth and ultrastructure were analysed as well as their cytotoxicity to murine macrophages. Both oils were highly active with IC50/96 h of 8.69 and 9.53 µg/mL for M. ovata and E. erythropappus against promastigotes and caused ultrastructural alterations including mitochondrial enlargement. Cytotoxicity for murine macrophages varied with the oil concentrations. The IC50 low values of both M. ovata and E. erythropappus oils against L. amazonensis and their relative low cytotoxicity to mammal host cells support their potential use against cutaneous leishmaniasis.

Development and characterization of nuclear microsatellite markers for Eremanthus erythropappus and their transferability across related species.

BACKGROUND: We developed simple sequence repeats (SSR) for Eremanthus erythropappus (DC.) MacLeish, an endangered tree species endemic to the Brazilian Savanna and Atlantic Forest biomes, and tested their transferability to two closely related Eremanthus species. RESULTS: Using a genomic library enriched with tandem repeat motifs, we identified 16 primer pairs, and characterized them in two populations. Nine primers amplified the expected size fragments and seven SSRs were polymorphic, providing a total of 38 alleles and an average of 4.22 alleles per marker. The polymorphic information content (PIC) ranged from 0.44 to 0.94 with an average of 0.65. The average observed heterozygosity across all loci varied from 0.61 to 1.00. The observed (HO) and expected (HE) heterozygosity within the two populations varied from 0.65 to 1.00 and from 0.31 to 1.00, respectively. CONCLUSIONS: These newly developed SSR markers are a powerful tool for population genetic analyses and may be useful in studies on species ecology, evolution, and taxonomy.

Development and characterization of nuclear microsatellite markers for Eremanthus erythropappus and their transferability across related species

BACKGROUND: We developed simple sequence repeats (SSR) for Eremanthus erythropappus (DC.) MacLeish, an endangered tree species endemic to the Brazilian Savanna and Atlantic Forest biomes, and tested their transferability to two closely related Eremanthus species. RESULTS: Using a genomic library enriched with tandem repeat motifs, we identified 16 primer pairs, and characterized them in two populations. Nine primers amplified the expected size fragments and seven SSRs were polymorphic, providing a total of 38 alleles and an average of 4.22 alleles per marker. The polymorphic information content (PIC) ranged from 0.44 to 0.94 with an average of 0.65. The average observed heterozygosity across all loci varied from 0.61 to 1.00. The observed ( HO ) and expected ( HE ) heterozygosity within the two populations varied from 0.65 to 1.00 and from 0.31 to 1.00, respectively. CONCLUSIONS: These newly developed SSR markers are a powerful tool for population genetic analyses and may be useful in studies on species ecology, evolution, and taxonomy.

Identification of the Bisabolol Synthase in the Endangered Candeia Tree (Eremanthus erythropappus (DC) McLeisch).

Candeia (Eremanthus erythropappus (DC) McLeisch, Asteraceae) is a Brazilian tree, mainly occurring in the cerrado areas. From ethnobotanical information its essential oil is known to have wound healing and nociceptive properties. These properties are ascribed to result from a sesquiterpene alcohol, (-)-α-bisabolol, which is present at high concentrations in this oil. Bisabolol is highly valued by the cosmetic industry because of its antibacterial, anti-inflammatory, skin-smoothing and wound healing properties. Over the past decades, Candeia timber has been collected at large scale for bisabolol extraction from wild reserves and the species is thereby at risk of extinction. To support the development of breeding and nursing practices that would facilitate sustainable cultivation of Candeia, we identified a terpene synthase gene, EeBOS1, that appears to control biosynthesis (-)-α-bisabolol in the plant. Expression of this gene in E. coli showed that EeBOS1 protein is capable of producing (-)-α-bisabolol from farnesyl pyrophosphate in vitro. Analysis of gene expression in different tissues from Candeia plants in different life stages showed a high correlation of EeBOS1 expression and accumulation of (-)-α-bisabolol. This work is the first step to unravel the pathway toward (-)-α-bisabolol in Candeia, and in the further study of the control of (-)-α-bisabolol production.

Influence of soil texture on nutrients and potentially hazardous elements in Eremanthus erythropappus.

Understanding the factors that control uptake rates and allocation of chemical elements among plant organs is a fundamental prerequisite to improve phytostabilization techniques of hazardous elements in contaminated areas. The present study shows evidence that different substrate textures (coarse and fine laterite) do not significantly change the partitioning of root and shoot dry biomass and with few exceptions, do not significantly affect the final average concentration of elements in Eremanthus erythropappus, but change the root:shoot allocation of both essential nutrients and elements potentially toxic to biota. Growth on coarse laterite resulted in significant higher K (30%), Mg (34%), P (25%), S (32%), Cu (58%), and Na (43%) concentrations in roots and lower Cd concentration (29%). In shoots, coarse laterite led to reduction in K, Fe, Al, and Cr and increase in Na and Sr concentrations. Changes in element allocation could be, in part, a result of differences in the water availability of substrates. Matric potential in coarse laterite was significantly lower in at least 47% of the days analyzed throughout the year. Changes in element phytoextraction or phytostabilization potential could influence the efficiency of rehabilitation projects in areas degraded by mining activities.

Assessing the Chemical Composition and Antimicrobial Activity of Essential Oils from Brazilian Plants-Eremanthus erythropappus (Asteraceae), Plectrantuns barbatus, and P. amboinicus (Lamiaceae).

The chemical composition and antimicrobial activity of essential oils obtained from three Brazilian plant species-leaves and branches of Eremanthus erythropappus (Asteraceae), leaves of Plectranthus barbatus, and leaves of P. amboinicus (Lamiaceae)-were determined. Analysis by GC/MS and determination of Kovats indexes both indicated δ-elemene (leaves-42.61% and branches-23.41%) as well as (-)-α-bisabolol (leaves-24.80% and stem bark-66.16%) as major constituents of E. erythropappus essential oils. The main components of leaves of P. barbatus were identified as (Z)-caryophyllene (17.98%), germacrene D (17.35%), and viridiflorol (14.13%); whereas those of leaves of P. amboinicus were characterized as p-cymene (12.01%), γ-terpinene (14.74%), carvacrol (37.70%), and (Z)-caryophyllene (14.07%). The antimicrobial activity against yeasts and bacteria was assessed in broth microdilution assays to determine the minimum inhibitory concentration (MIC) necessary to inhibit microbial growth. In addition, the crude oil of branches of E. erythropappus was subjected to chromatographic separation procedures to afford purified (-)-α-bisabolol. This compound displayed biological activity against pathogenic yeasts, thus suggesting that the antimicrobial effect observed with crude oils of E. erythropappus leaves and branches may be related to the occurrence of (-)-α-bisabolol as their main component. Our results showed that crude oils of Brazilian plants, specifically E. erythropappus, P. barbatus, and P. amboinicus and its components, could be used as a tool for the developing novel and more efficacious antimicrobial agents.

Growing Eremanthus erythropappus in crushed laterite: A promising alternative to topsoil for bauxite-mine revegetation.

Topsoil is the preferred substrate for areas requiring rehabilitation after bauxite mining. However, topsoil is sometimes lacking and so there is a need to test the suitability of other, locally available substrates. In an abandoned bauxite mine in Southeastern Brazil, small patches of native vegetation spontaneously established in shallow depressions over weathered laterite, suggesting that granulometric reduction may have facilitated the establishment of plants. To test this hypothesis, blocks of laterite collected in the area were crushed to simulate texture observed in the vegetation patches. Topsoil collected in a preserved ferruginous field near to the extraction area was also used as a substrate in which Eremanthus erythropappus seedlings, a native woody species, were grown. Seedlings were cultivated without fertilizers in these two substrates and also directly over the exposed and uncrushed laterite. The species proved to be very promising for the revegetation, showing a high survival rate in all substrates. Higher annual growth rates and higher final biomass values were observed in topsoil, but the granulometric reduction of laterite doubled plant growth rate in comparison to the exposed laterite. This result was likely due to the increased availability of essential nutrients to plants and to the improvement in physical conditions for root growth and functioning. Moreover, seedling allometry was not altered by the type of substrate, suggesting that the species was highly tolerant to the new substrate conditions, a fundamental characteristic for success of revegetation of bauxite extraction degraded areas.

Caracterização morfoanatômica das folhas de Eremanthus erythropappus (DC.) MacLeisch, Asteraceae / Leaf morpho-anatomical characterization of Eremanthus erythropappus (DC.) MacLeisch, Asteraceae

Eremanthus erythropappus (DC.) MacLeisch (Asteraceae), popularmente conhecida como "candeia-da-serra", é uma espécie utilizada na medicina tradicional como cicatrizante e antimicrobiano. O objetivo do presente trabalho foi comparar o número de tricomas glandulares nas folhas jovens e adultas de E. erythropappus, assim como realizar a caracterização morfoanatômica destas folhas. Secções transversais e paradérmicas das folhas foram submetidas às microtécnicas fotônicas e à microscopia eletrônica de varredura. A quantificação dos tricomas glandulares foi realizada em folhas jovens e adultas com auxílio de câmara clara. As folhas são alternas ou fasciculadas e a lâmina foliar possui epiderme uniestratificada, revestida por cutícula delgada e lisa e mesofilo dorsiventral. Em ambas as faces da epiderme, ocorrem estômatos predominantemente anomocíticos e tricomas glandulares inseridos em depressões. Na face abaxial observam-se numerosos tricomas tectores. O parênquima paliçádico é uniestratificado e o parênquima voltado para a face abaxial é formado por três a cinco camadas de células com disposição compacta. As folhas jovens e adultas apresentam respectivamente 21,78±5,83 e 17,80±6,69 tricomas glandulares na face adaxial. A análise morfoanatômica das folhas de E. erythropappus mostra-se um método rápido e prático para a identificação e controle de qualidade de espécies vegetais utilizadas na terapêutica.

Eremanthus erythropappus (DC.) MacLeisch (Asteraceae), commonly known as 'candeia-da-serra', is a plant used in folk medicine as wound healing and antimicrobial. The aim of this study was to compare the number of glandular trichomes between the young and the mature leaves, as well as to perform the morpho-anatomical characterization of E. erythropappus leaves. Transverse and paradermal sections of the leaves were prepared according to light and scanning microtechniques for the morpho-anatomical characterization. The quantification of glandular trichomes on the adaxial surface of the epidermis was evaluated in young and mature leaves with camera lucida. The leaves are alternate or fasciculate and the blade has uniseriate epidermis coated with thin and smooth cuticle and dorsiventral mesophyll. There are predominantly anomocytic stomata on both surfaces, as well as glandular trichomes located in epidermal depressions. Various non-glandular trichomes are encountered on the abaxial surface. The palisade parenchyma consists of a single layer of cells and the parenchyma which is faced to the abaxial surface comprehends three to five layers of cells in compact arrangement. The young and mature leaves showed, respectively, 21.78±5.83 e 17.80±6.69 glandular trichomes on the adaxial side. The morpho-anatomical analysis of E. erythropappus leaves has proved to be a practical and rapid method for the identification and quality control of the vegetal species used for medical purposes.

PRODUCCIÓN Y CARACTERIZACIÓN DE CARBONES ACTIVADOS A PARTIR DE RESIDUOS DE CANDEIA (Eremanthus erythropappus) Y SU APLICACIÓN EN LA ADSORCIÓN DE COMPUESTOS ORGÁNICOS / PRODUCTION AND CHARACTERIZATION OF ACTIVATED CARBONS FROM WASTE CANDEIA (Eremanthus erythropappus) AND THEIR APLICATION ON ORGANIC COMPOUNDS ADSORPTION / PRODUÇÃO E CARACTERIZAÇÃO DE CARVÕES ATIVADOS A PARTIR DE RESIDUOS DE CANDEIA (Eremanthus erythropappus) E TESTES DE ADSORÇÃO DE compostos orgânicos

En este trabajo se prepararon dos carbones activados (CA), utilizando como agentes activantes ZnCl2 yK2CO3, denominados CA/ZnCl2,CA/K2CO3, respectivamente, a partir de residuos generados en el proceso de extracción del aceite esencial de candeia, árbol nativo de Brasil. Estos carbones se caracterizaron por microscopía electrónica de barrido, análisis termogravimétrico, análisis elemental, superficie específica por el método BET e índice de iodo. Estos materiales se probaron en la adsorción de dos moléculas modelos: el fenol y el azul de metileno, y se comparó su capacidad de adsorción con un carbón activado comercial (Merck), denominado CA/Com. Las isotermas de adsorción para CA/ZnCl2, CA/K2CO3 y CA/Com mostraron capacidades de adsorción máxima de 297, 228 y 271 mg g-1, para azul de metileno y de 109, 195 y 161 mg g-1, para el fenol, respectivamente. Los resultados evidenciaron que los carbones son eficientes en el proceso de adsorción de las moléculas modelo ensayadas. Las isotermas fueron ajustadas a las ecuaciones de Langmuir y de Freundlich. El modelo de Langmuir entregó un ajuste mejor, mostrando que los adsorbentes presentan superficies energéticamente más uniformes. Los carbones activados producidos se presentan como potenciales adsorbentes para compuestos orgánicos, agregando un valor adicional al residuo.

In this work, two activated carbons were prepared, using as activating agents ZnCl2 and K2CO3, denominated CA/ZnCl2,CA/ K2CO3, respectively, from waste generated in the process of extraction of essential oil of Candeia, a tree native of Brazil. These carbons were characterized by scanning electron microscopy, thermal analysis, elemental analysis, BET surface area and iodine index. These materials were tested in the adsorption of model molecules: Phenol and Methylene Blue, and a commercial activated carbon (Merck) was used for comparison purposes. Adsorption isotherms for CA/ZnCl2,CA/K2CO3 and CA/Com showed maximum sorption capacities of 297, 228 and 271 mg g-1 for Methylene Blue and 109,195 and 161 mg g-1 for phenol respectively. The results showed that the carbons are efficient in the adsorption process of molecules tested. The isotherms were fitted to equations of Langmuir and Freundlich. The Langmuir model provided better fit, showing that the adsorbents have energetically uniform surfaces. The materials produced were characterized as potential adsorbents for organic compounds that can be used as an alternative for obtaining activated carbons adding value to the residue.

Neste trabalho, foram preparados dois carvões ativados, utilizando como agentes ativantes o ZnCl² eoK2CO3, a partir de resíduos gerados no processo de extração de óleo essencial de Candeia, árvore nativa no Brasil. Estes carvões foram caracterizados por microscopia eletrônica de varredura, análise termo-gravimétrica, análise elementar, área superficial BET e índice de iodo. E testados na Adsorção das moléculas modelo: Fenol e Azul de Metileno, foi utilizado um carvão comercial (Merck) para fins de comparação. As isotermas de Adsorção para o CA/ZnCl2, CA/K2CO3 e CA/Com apresentaram capacidades máximas de adsorção de 297, 228 e 271 mg g-1 para Azul Metileno e de 109, 195 e 161 mg g-1 para fenol respectivamente. Os resultados evidenciaram que os carvões são eficientes no processo de adsorção das moléculas testadas. As isotermas foram ajustadas as equações de Langmuir e Freundlich, o modelo de Langmuir forneceu melhor ajuste, mostrando que os ad-sorventes apresentam superfícies energeticamente uniformes. Os materiais produzidos se caracterizaram como potenciais adsorventes para compostos orgânicos podendo ser utilizados como alternativa para obtenção de carvões ativados, além de agregar valor ao resíduo.

Propriedades farmacológicas do extrato etanólico de Eremanthus erythropappus (DC. ) McLeisch (Asteraceae) / Pharmacological properties of the ethanol extract from Eremanthus erythropappus (DC. ) McLeisch (Asteraceae)

O presente trabalho investigou as propriedades antinociceptiva, antiinflamatória e antiulcerogênica do extrato etanólico dos galhos de E. erythropappus. Foram realizados os testes de contorções abdominais induzidas por ácido acético; tempo da lambida da pata induzida por formalina; placa quente; edema de pata induzido por carragenina; lesões ulcerativas induzidas por indometacina e etanol. As doses de 50, 100 e 200 mg/kg inibiram as contorções em 43,07 por cento, 69,34 por cento e 70,07 por cento, respectivamente. O tempo da lambida da pata foi reduzido em ambas as fases nas doses testadas, enquanto o tempo de latência da placa quente teve efeito significativo na dose de 200 mg/kg. O edema de pata foi reduzido em 23,81 por cento (100 mg/kg) e 47,62 por cento (200 mg/kg). O índice ulcerativo das lesões induzidas por indometacina foi reduzido na dose de 100 mg/kg e 200 mg/kg, enquanto a dose de 200 mg/kg teve efeito significativo sobre o volume e o pH do suco gástrico. O índice ulcerativo e o volume do suco gástrico, induzidos por etanol, foram reduzidos na dose de 200 mg/kg, enquanto o pH do suco gástrico aumentou nas doses de 100 e 200 mg/kg. Os resultados indicam que o extrato etanólico de E. erythropappus avaliado pode constituir alvo potencial para uso em terapias da dor, da inflamação e de úlcera.

The present work investigated the antinociceptive, anti-inflammatory and antiulcerogenic activities of Eremanthus erythropappus ethanol extract. The tests were carried out by acetic acid writhing, paw licking induced by formalin, hot plate, indomethacin- and ethanol-induced ulcer. The doses of 50, 100 and 200 mg/kg inhibited the contortions in 43.07, 69.34 and 70.07 percent, respectively. Both phases of paw lick were reduced at the tested doses, while reaction time at hot plate had significant effect at 200 mg/kg. The paw edema was decreased in 23.81 percent (100 mg/kg) and 47.62 percent (200 mg/kg). The index of ulceration induced by indomethacin was reduced at the doses of 100 and 200 mg/kg, while 200 mg/kg decreased the gastric volume and pH. The index of ulceration and gastric volume induced by ethanol was reduced at 200 mg/kg, while the pH increased at 100 and 200 mg/kg. The results indicate that the ethanol extract from E. erythropappus may constitute a potential target for the use in therapies of the pain, inflammation and ulcer.

Contrasting genetic diversity and differentiation of populations of two successional stages in a neotropical pioneer tree (Eremanthus erythropappus, Asteraceae)

Eremanthus erythropappus, commonly known as “candeia”, is an abundant pioneer tree species, forming dense populations known as “candeial”, but it is also found in forests at middle stages of succession. Trees from forests are bigger and occur in lower density than in the “candeial”. The objectives of the present study were to investigate if the decrease in population density during successional process is accompanied by 1) changes in within-population genetic diversity, and 2) differentiation of populations. Eight populations, four of early successional stage (“candeial”) and four of middle successional stages (forest), were analyzed with RAPD markers. The genetic diversity found was high compared to other tree species analyzed with RAPD markers. AMOVA revealed that most of the genetic variations of E. erythropappus were found within populations (85.7%), suggesting that this species is predominantly outcrossing. The relatively low differentiation among the populations can be attributed to small distances among the populations analyzed (0.2 to 10.8 km). No indication that populations from middle successional habitats show lower genetic variation than populations from early successional stages was found. The percentage of polymorphic fragments (82.8 and 84.8%) and the Shannon indexes (0.442 and 0.455) were similar in “candeial” and forest, respectively. These results suggest that if an increase in selection intensity occurred during succession, it did not result in a decrease in genetic diversity or that the selection effect was balanced by other factors, such as gene flow. Higher significant differentiation among E. erythropappus populations from “candeial” in relation to that among populations from forest was also not detected.