Morphophysiological, ultrastructural, and nutritional changes induced by Cu toxicity in young Erythrina fusca plants.

Erythrina fusca is an important legume used for shade cover in cacao plantations in Brazil. Cacao plantations receive large quantities of copper (Cu)-containing agrochemicals, mainly for control of diseases. Therefore, Cu toxicity was investigated in seedlings grown in hydroponics with increasing concentrations of Cu (0.005-32 mg L-1) in a greenhouse. Ultrastructural analyses showed cell plasmolysis in the root cortical area and changes in thylakoid membranes at 8 mg Cu L-1 and higher. There were changes in epicuticular wax deposition on the leaf surface at the 16 and 32 mg Cu L-1 treatments. Leaf gas exchanges were highly affected 24 hours after application of treatments beginning at 8 mg Cu L-1 and higher Cu concentrations. Chemical analyses showed that Cu content in E. fusca roots increased as Cu concentration in the nutrient solution increased, whereas the shoot did not show significant changes. It is also observed that excess Cu interfered with Zn, Fe, Mn, Mg, K, P, and Ca content in the different E. fusca organs. Investigation of Cu toxicity symptoms focusing on morphophysiological, ultrastructural, gas exchange, and nutritional changes would be useful to alleviate Cu toxicity in E. fusca under field conditions, an important agroforestry species in cacao plantation.

Making a Bad Situation Worse: An Invasive Species Altering the Balance of Interactions between Local Species.

Biological invasions pose a significant threat to biodiversity, especially on oceanic islands. One of the primary explanations for the success of plant invaders is direct suppression of competitors. However, indirect interactions can also be important, although they are often overlooked in studies on biological invasion. The shrub Leucaena leucocephala is a widespread island invader with putative allelopathic effects on the germination and growth of other species. We quantified the impact of Leucaena on plant communities richness on an oceanic Brazilian island and, through nursery experiments, investigated the potential for allelopathic effects on the germination of Erythrina velutina, a native species that is often absent from stands of Leucaena. Additionally, in a manipulative field experiment, we examined the direct and indirect effects (mediated by the native species Capparis flexuosa) of the invader on the development of Erythrina. The species richness in invaded sites was lower than in uninvaded sites, and Capparis was the only native species that was frequently present in invaded sites. In the nursery experiments, we found no evidence that Leucaena affects the germination of Erythrina. In the field experiments, the odds of Erythrina germination were lower in the presence of Leucaena litter, but higher in the presence of Leucaena trees. However, the survival and growth of Erythrina were considerably inhibited by the presence of Leucaena trees. The isolated effect of native Capparis on the germination and growth of Erythrina varied from positive to neutral. However, when Capparis and Leucaena were both present, their combined negative effects on Erythrina were worse than the effect of Leucaena alone, which may be attributed to indirect effects. This study provides the first empirical evidence that the balance of the interactions between native species can shift from neutral/positive to negative in the presence of an exotic species.

Dormancy-breaking requirements of Sophora tomentosa and Erythrina speciosa (Fabaceae) seeds.

The physical dormancy of seeds has been poorly studied in species from tropical forests, such as the Atlantic Forest. This study aimed to examine the effect of moderate alternating temperatures on breaking the physical dormancy of seeds, the morphoanatomy and histochemistry of seed coats, and to locate the structure/region responsible for water entrance into the seed, after breaking the physical dormancy of seeds of two woody Fabaceae (subfamily Faboideae) species that occur in the Brazilian Atlantic Forest: Sophora tomentosa and Erythrina speciosa. To assess temperature effect, seeds were incubated in several temperature values that occur in the Atlantic Forest. For morphological and histochemical studies, sections of fixed seeds were subjected to different reagents, and were observed using light or epifluorescence microscopy, to analyze the anatomy and histochemistry of the seed coat. Treated and nonreated seeds were also analyzed using a scanning electron microscope (SEM) to observe the morphology of the seed coat. To localize the specific site of water entrance, the seeds were blocked with glue in different regions and also immersed in ink. In the present work a maximum temperature fluctuation of 15 degrees C was applied during a period of 20 days and these conditions did not increase the germination of S. tomentosa or E. speciosa. These results may indicate that these seeds require larger fluctuation of temperature than the applied or/and longer period of exposition to the temperature fluctuation. Blocking experiments water inlet combined with SEM analysis of the structures of seed coat for both species showed that besides the lens, the hilum and micropyle are involved in water absorption in seeds scarified with hot water. In seeds of E. speciosa the immersion of scarified seeds into an aniline aqueous solution showed that the solution first entered the seed through the hilum. Both species showed seed morphological and anatomical features for seed coats of the subfamily Faboideae. Lignin and callose were found around all palisade layers and the water impermeability and ecological role of these substances are discussed in the work.

Dormancy-breaking requirements of Sophora tomentosa and Erythrina speciosa (Fabaceae) seeds / Dormancy-breaking requirements of Sophora tomentosa and Erythrina speciosa (Fabaceae) seeds

The physical dormancy of seeds has been poorly studied in species from tropical forests, such as the Atlantic Forest. This study aimed to examine the effect of moderate alternating temperatures on breaking the physical dormancy of seeds, the morphoanatomy and histochemistry of seed coats, and to locate the structure/region responsible for water entrance into the seed, after breaking the physical dormancy of seeds of two woody Fabaceae (subfamily Faboideae) species that occur in the Brazilian Atlantic Forest: Sophora tomentosa and Erythrina speciosa. To assess temperature effect, seeds were incubated in several temperature values that occur in the Atlantic Forest. For morphological and histochemical studies, sections of fixed seeds were subjected to different reagents, and were observed using light or epifluorescence microscopy, to analyze the anatomy and histochemistry of the seed coat. Treated and non-treated seeds were also analyzed using a scanning electron microscope (SEM) to observe the morphology of the seed coat. To localize the specific site of water entrance, the seeds were blocked with glue in different regions and also immersed in ink. In the present work a maximum temperature fluctuation of 15ºC was applied during a period of 20 days and these conditions did not increase the germination of S. tomentosa or E. speciosa. These results may indicate that these seeds require larger fluctuation of temperature than the applied or/and longer period of exposition to the temperature fluctuation. Blocking experiments water inlet combined with SEM analysis of the structures of seed coat for both species showed that besides the lens, the hilum and micropyle are involved in water absorption in seeds scarified with hot water. In seeds of E. speciosa the immersion of scarified seeds into an aniline aqueous solution showed that the solution first entered the seed through the hilum. Both species showed seed morphological and anatomical features for seed coats of the subfamily Faboideae. Lignin and callose were found around all palisade layers and the water impermeability and ecological role of these substances are discussed in the work.

Requisitos para romper la latencia en semillas de Sophora tomentosa y Erythrina speciosa (Fabaceae). La latencia física de las semillas ha sido poco estudiada en las especies de los bosques tropicales, como el bosque atlántico. Este estudio tuvo como objetivo examinar el efecto de las temperaturas moderadas alternantes en romper la latencia física de las semillas, la anatomía y la histoquímica de la cubierta de las semillas, y la localización de la estructura o región responsable de la entrada de agua, después de romper la latencia física de las semillas de Sophora tomentosa y Eythrina speciosa, dos especies leñosas de Fabaceae (subfamilia Faboideae) que presentes en el bosque atlántico de Brasil. Para cumplir con el primer objetivo se incubaron las semillas a varias temperaturas que se dan en el bosque atlántico. Para los estudios morfológicos e histoquímicos se fijaron secciones de semillas sometidos a diferentes reactivos y se observaron usando luz o microscopía de epifluorescencia para analizar la anatomía y la histoquímica de la cubierta de la semilla. Semillas tratadas y no tratadas se analizaron también usando un microscopio electrónico de barrido (MDB) o microscopio estereoscópico (ME) para observar la morfología de la cubierta de la semilla. Para localizar el sitio específico de la entrada de agua, las semillas fueron bloqueadas con pegamento en diferentes regiones y también sumergidas en tinta. En el presente trabajo se aplicó una fluctuación de temperatura máxima de 15°C durante un período de 20 días y estas condiciones no aumentó la germinación de S. tomentosa o E. speciosa. Estos resultados pueden indicar que estas semillas requieren mayor fluctuación de la temperatura que la aplicada y/o un período más largo de exposición a la fluctuación de la temperatura. Experimentos de bloqueo de entrada de agua combinada con el análisis de las estructuras de la cubierta de la semilla para ambas especies (SEM) mostró que, a pesar de la lente, el hilio y micropilo están implicados en la absorción de agua en las semillas escarificadas con agua caliente. En las semillas de E. speciosa la inmersión de semillas escarificadas en una solución acuosa de anilina mostró que la solución entró por primera vez a la semilla a través del hilio. Ambas especies mostraron características morfológicas y anatómicas de semillas con cubierta de la subfamilia Faboideae. La lignina y callosa se encontraron alrededor de todas las capas de empalizada y la impermeabilidad al agua y en el trabajo se discute el papel ecológico de estas sustancias.

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.

Propagation and conservation of native forest genetic resources of medicinal use by means of in vitro and ex vitro techniques.

In Argentina, there are numerous native species which are an important source of natural products and which are traditionally used in medicinal applications. Some of these species are going through an intense extraction process in their natural habitat which may affect their genetic diversity. The aim of this study was to establish vegetative propagation systems for three native forestal species of medicinal interest. This will allow the rapid obtainment of plants to preserve the germplasm. This study included the following species which are widely used in folk medicine and its applications: Erythrina crista-galli or "seibo" (astringent, used for its cicatrizant properties and for bronchiolitic problems); Acacia caven or "espinillo" (antirheumatic, digestive, diuretic and with cicatrizant properties) and Salix humboldtiana or "sauce criollo" (antipyretic, sedative, antispasmodic, astringent). The methodology included the micropropagation of seibo, macro and micropropagation of Salix humboldtiana and the somatic embryogenesis of Acacia caven. The protocol for seibo regeneration was adjusted from nodal sections of seedlings which were obtained from seeds germinated in vitro. The macropropagation through rooted cuttings of "sauce criollo" was achieved and complete plants of this same species were obtained through both direct and indirect organogenesis using in vitro cultures. The somatic embryogenesis for Acacia caven was optimized and this led to obtain a high percentage of embryos in different stages of development. We are able to support the conservation of native forest resources of medicinal use by means of vegetative propagation techniques.

Primeiro registro no Brasil de Erythrina velutina Willd. como hospedeira de Tetranychus neocaledonicus (Acari: Tetranychidae) / First Brazilian report of Erythrina velutina Willd. as host of Tetranychus neocaledonicus (Acari: Tetranychidae)

Os ácaros tetraniquídeos representam importantes pragas para diversas culturas agrícolas em todo o mundo. Durante a prospecção em mudas de Erytrina velutina no distrito de São Cristovão, Estado de Sergipe, Brasil, a ocorrência destes foi observada em 15 por cento das mudas que foram levadas ao laboratório para a caracterização dos danos. As mudas apresentavam sintomas de redução e encarquilhamento do limbo. Foram coletados 100 indivíduos destas plantas, os quais foram montados em lâminas de microscopia para a identificação taxonômica. Os ácaros foram identificados como pertencentes à espécie Tetranychus neocaledonicus (Acari: Tetranychidae). Este é o primeiro relato em E. velutina, como hospedeira para T. neocaledonicus no Brasil.

Spider mites are important pests to several crops worldwide. During prospecting in Erytrina velutina specimens from São Cristovão District, Sergipe State, Brazil, their occurrence was observed in 15 percent seedlings, which were taken to the laboratory for description of damages. The seedlings presented leaf blade reduction and crumpling symptoms. One hundred mites were collected from these seedlings and mounted on microscope slides for taxonomic identification. The identified mites belonged to the species Tetranychus neocaledonicus (Acari: Tetranychidae). This is the first report of E. velutina as host for T. neocaledonicus in Brazil.

Erythrina speciosa (Leguminosae-Papilionoideae) under soil water saturation: morphophysiological and growth responses.

BACKGROUND AND AIMS: Erythrina speciosa is a Neotropical tree that grows mainly in moist habitats. To characterize the physiological, morphological and growth responses to soil water saturation, young plants of E. speciosa were subjected experimentally to soil flooding. METHODS: Flooding was imposed from 2 to 4 cm above the soil surface in water-filled tanks for 60 d. Non-flooded (control) plants were well watered, but never flooded. The net CO(2) exchange (A(CO2)), stomatal conductance (g(s)) and intercellular CO(2) concentration (C(i)) were assessed for 60 d. Soluble sugar and free amino acid concentrations and the proportion of free amino acids were determined at 0, 7, 10, 21, 28 and 45 d of treatments. After 28, 45 and 60 d, dry masses of leaves, stems and roots were determined. Stem and root cross-sections were viewed using light microscopy. KEY RESULTS: The A(CO2) and g(s) were severely reduced by flooding treatment, but only for the first 10 d. The soluble sugars and free amino acids increased until the tenth day but decreased subsequently. The content of asparagine in the roots showed a drastic decrease while those of alanine and gamma-aminobutyric increased sharply throughout the first 10 d after flooding. From the 20th day on, the flooded plants reached A(CO2) and g(s) values similar to those observed for non-flooded plants. These events were coupled with the development of lenticels, adventitious roots and aerenchyma tissue of honeycomb type. Flooding reduced the growth rate and altered carbon allocation. The biomass allocated to the stem was higher and the root mass ratio was lower for flooded plants when compared with non-flooded plants. CONCLUSIONS: Erythrina speciosa showed 100 % survival until the 60th day of flooding and was able to recover its metabolism. The recovery during soil flooding seems to be associated with morphological alterations, such as development of hypertrophic lenticels, adventitious roots and aerenchyma tissue, and with the maintenance of neutral amino acids in roots under long-term exposure to root-zone O(2) deprivation.

Phytodegradation potential of Erythrina crista-galli L., Fabaceae, in petroleum-contaminated soil.

This work aimed at investigating both the tolerance and the phytodegradation potential of Erythrina crista-galli L. in petroleum-contaminated soil. It consisted in analyzing E. crista-galli germination, surviving, growth, and development when cultivated at different contaminant concentrations and pollutant degradation rates. This specimen was selected because it presented a special behavior among others also exposed to petroleum in an accident that occurred in the Araucaria region (south of Brazil), resulting in a four-million-liter oil spill. The experiment was carried out in a greenhouse containing non-contaminated soil (NCS), vegetated contaminated soil (VCS), and non-vegetated contaminated soil (NVCS) at the following petroleum concentrations: 25 g kg(-1) (VCS-25), 50 g kg(-1) (VCS-50), and 75 g kg(-1) (VCS-75). After 60 days, the soil samples were analyzed by gas chromatography. Germination was more and more evident as higher petroleum concentrations were observed. The surviving rates of groups NCS, VCS-25, VCS-50, and VCS-75 were 64%, 70%, 61%, and 96%, respectively. The VCS group growth was reduced when compared to the control group (NCS). The individuals exposed to petroleum pollution presented differences in the anatomic structure of their roots when compared to the NCS group. It was observed that the petroleum degradation rate was higher for VCS group than for NVCS. E. crista-galli is potentially recommended for petroleum-contaminated soils because of its positive association in the presence of contamination.

Production of alkaloids by in vitro culture of Erythrina americana Miller.

The production of erythroidines and other alkaloids was studied in cotyledons, callus and cell suspension cultures of Erythrina americana Miller. The cell suspension cultures, grown in Murashige & Skoog medium with naphthaleneacetic acid (3 mg l(-1)) and kinetin (2 mg l(-1)), produced 89 and 17 microg alpha- and beta-erythroidines respectively per g dry wt.