A NOVEL EPIPHYTIC CYANOBACTERIAL SPECIES FROM THE GENUS BRASILONEMA CAUSING DAMAGE TO EUCALYPTUS LEAVES

A cyanobacterial mat colonizing the leaves of Eucalyptus grandis was determined to be responsible for serious damage affecting the growth and development of whole plants under the clonal hybrid nursery conditions. The dominant cyanobacterial species was isolated in BG-11 medium lacking a source of combined nitrogen and identified by cell morphology characters and molecular phylogenetic analysis (16S rRNA gene and cpcBA-IGS sequences). The isolated strain represents a novel species of the genus Brasilonema and is designated Brasilonema octagenarum strain UFV-E1. Thin sections of E. grandis leaves analyzed by light and electron microscopy showed that the B. octagenarum UFV-E1 filaments penetrate into the leaf mesophyll. The depth of infection and the mechanism by which the cyanobacterium invades leaf tissue were not determined. A major consequence of colonization by this cyanobacterium is a reduction in photosynthesis in the host since the cyanobacterial mats decrease the amount of light incident on leaf surfaces. Moreover, the cyanobacteria also interfere with stomatal gas exchange, decreasing CO2 assimilation. To our knowledge, this is the first report of an epiphytic cyanobacterial species causing damage to E. grandis leaves.

Effect of eucalyptus (Eucalyptus citriodora) fresh or residue leaves on methane emission in vitro

Rumen fermentation and methane emission for eucalyptus (Eucalyptus citriodora) fresh leaves (FL) or residue leaves (RL), after essential oil extraction from eucalyptus leaves in comparison with alfalfa (Medicago sativa) hay, were investigated in vitro. Eucalyptus FL and RL were obtained from the Distillery Trees Barras Company, Torrinha City, Sao Paulo, Brazil. The semi-automatic system of gas production was used to measure gas production, methane emission and rumen fermentation after 24 h incubation in vitro. The results showed that the crude protein (CP) contents were 76.4, 78.1 and 181.9 g kg(-1) DM for eucalyptus FL, RL and alfalfa hay, respectively. The neutral-detergent fibre (NDF) and acid-detergent fibre (ADF) were significantly lower in eucalyptus FL and RL than alfalfa hay. The Eucalyptus fresh and residue leaves were rich in total phenols (TP) and total tannins (TT) but had negligible content of condensed tannins (CT). There was significant reduction in cumulative gas production about 54 and 51% with eucalyptus FL and RL, respectively, compared with alfalfa hay. The methane emission (mL/g DM) was reduced (P<0.05) by 53 and 57% with eucalyptus FL and RL, respectively, but the reduction was 21 and 16% when expressed on truly digested organic matter basis. There were a decline (P<0.05) in true dry and organic matter degradation in vitro in eucalyptus FL and RL compared with alfalfa hay substrate. The partitioning factor values were higher (P<0.05) in eucalyptus FL and RL than alfalfa hay. There was no significant difference observed between eucalyptus FL, RL and alfalfa hay in protozoa count. It is concluded that the eucalyptus leaves have potential effect to mitigate CH4 production in vitro, which may be attributed to a decrease in fermentable substrate rather than to a direct effect on methanogenesis.

The Allelopathic Effect of Eucalyptus Leaf Extract on Grass Forage Seed

This study aimed to evaluate the allelopathic effect of various concentrations of an aqueous extract of eucalyptus leaves on Urochloa decumbens and Panicum maximum seeds. The extract was prepared from Eucalyptus urograndis leaves that were milled and mixed with distilled water in a 1:9 milled leaves: water ratio to obtain an extract with a defined concentration of 100%. In addition, dilutions of 50%, 25% and 12.5% were prepared, and a 0% dilution was used as a control. The experiment followed a completely randomized design, with four replicates, each of 50 seeds of U. decumbens and 50 seeds of P. maximum, arranged on filter paper moistened with each concentration of extract in a Gerbox plastic box. The results demonstrated the allelopathic potential of E. urograndis aqueous extracts applied to the seeds of U. decumbens and P. maximum. The 50% and 100% concentrations of leaf extract most strongly inhibited the germination, vigor and seedling growth of U. decumbens and P. maximum. The germination speed index and the root length were the characteristics that were most affected by the potentially allelopathic substances contained in the eucalyptus extracts at all concentrations.

Otimização da extração sólido-líquido de antioxidantes de subprodutos florestais pelo método de superfície de resposta

Portugal continental apresenta uma vasta área florestal, que representa cerca de 35,4% da ocupação total do solo, com predominância de espécies como o eucalipto (Eucalyptus globulus) e o pinheiro-bravo (Pinus pinaster). Estas espécies apresentam uma elevada importância a nível económico, designadamente devido à sua ampla utilização, nomeadamente na indústria de celulose e papel, gerando elevadas quantidades de resíduos. Este resíduo de biomassa florestal é utilizado, na sua totalidade, para a geração de energia, na forma de eletricidade ou aquecimento. No entanto, existem outras opções viáveis, a nível económico, tais como a valorização destes subprodutos como fonte de compostos polifenólicos tornando-os, assim, um produto de valor acrescentado. A extração de compostos fenólicos de subprodutos florestais, como folhas de eucalipto e agulhas de pinheiros tem vindo a aumentar devido, principalmente, à substituição de antioxidantes sintéticos, contribuindo para a valorização de subprodutos florestais. Contudo, apesar de todas as potenciais aplicações e vantagens, apenas algumas centenas de espécies aromáticas identificadas são utilizadas à escala comercial. Neste trabalho foi avaliada a capacidade antioxidante de subprodutos da floresta, otimizando as condições de extração através do estudo dos fatores: tempo de extração, temperatura e composição de solvente através do método de superfície de resposta. O planeamento experimental utilizado teve como base um planeamento de compósito central e a avaliação do perfil de antioxidantes das matrizes analisadas foi realizada através de métodos de quantificação total, como o teor fenólico total, a atividade anti-radicalar – método do DPPH (radical 2,2-difenil-1-picrilhidrazilo) e o método de FRAP. Estes métodos analíticos convencionais foram modificados e, devidamente validados, para a análise em leitor de microplacas. Verificou-se que os extratos de pinheiro e de eucalipto, tanto as amostras verdes com as amostras, apresentam uma promissora capacidade antioxidante. O planeamento fatorial aplicado permitiu otimizar as condições de extração em relação às matrizes verdes. Contudo, o mesmo não se verificou em relação às matrizes secas. A composição (% de água) é sem dúvida o fator com mais efeito em todas as amostras (coeficientes de primeira e segunda ordem no modelo). Também a temperatura foi identificada como um fator com efeito significativo sobre os sistemas em análise.

Evaluation of the effect of the seed extract of Moringa oleifera Lam over the efficiency of organic filters in wastewater treatment of dairy cattle breeding

This study aimed to evaluate the effect of Moringa oleifera Lam extract on the removal of total solids (TS), total suspended solids (TSS) and chemical oxygen demand (COD), in different filter media for treating wastewater of dairy cattle breeding (DCW). The moringa seed extract was obtained by grinding 50 g of seeds in one liter of distilled water and, after passing the solution through a quantitative paper filter of 25 microns, 60 mL of the extract were added to wastewater from cattle breeding before the filtration process in organic filters made of thin coal, bamboo leaves, eucalyptus leaves, gliricidia branches and sawdust. This was followed by the completely randomized experimental design, adopting a factorial of 5 x 2. Aliquots of filtered effluent were collected and the total solids (TS) concentrations, total suspended solids (TSS) and chemical oxygen demand (COD) were determined. It was found that the increase in the efficiency of removal of COD and total solids can be attributed to the coagulating power of the moringa seed extract, wherein the filter medium with bamboo leaves presented the best performance, showing potential for use as alternative filter material in the primary treatment of DCW.

Amostragem e dinâmica populacional do percevejo bronzeado Thaumastocoris peregrinus (hemiptera: thaumastocoridae) em floresta clonal de eucalipto

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Caracterização morfológica e fenotípica de leveduras do gênero Cryptococcus com ênfase na pesquisa de Cryptococcus neoormans isoladas de amostras de água de mar e areia de três ambientes costeiros do estado de São Paulo: Canal de São Sebastião, Baixada Santista e Ubatuba

The yeast Cryptococcus neoformans is the etiologic agent of cryptococcosis, an infectious cosmopolitan disease that affects humans. Although rare, this disease is potentially fatal, especially for immunocompromised hosts. This pathogen is frequently isolated from excrements of pigeons and parrots, with many environmental sources such as birds, pigeon droppings, eucalyptus leaves, decaying trees, towers, churches and places of storage of grain (the port area). The isolation of this microorganism has been obtained also from the aquatic environment. The identification of environmental sources is needed to protect human health, especially susceptible populations such as immunocompromised. Therefore, this study investigated the presence of Cryptococcus neoformans in yeast isolates obtained from samples of sea water and sand from three regions of São Paulo: São Sebastião Channel, Santos and Ubatuba. Isolates were analyzed according to micro-and macroscopic characteristics and biochemical tests: microculture, urease, ink nankin, auxanograma, zymogram and phenol. We analyzed 199 isolates, 175 of which had features suggestive for Cryptococcus spp. in microculture. All these 175 isolates were sown in the Christensen urea middle to verify the production of urease and submitted to the technique nankin ink to visualize the capsule. Of these, only 24 were selected for the next test that was the auxanograma (assimilation of carbohydrate and nitrogen). Of the 24, 10 were tested in zymograms (fermented sugar), from which 5 were selected for the phenoloxidase test in medium containing dopamine. None of the 5 isolates tested had black or brown color characteristic of Cryptococcus neoformans. According to these tests, we arrived at 5 isolates identified to the genus Cryptococcus, but not the neoformans specie

Extratos vegetais na germinação de sementes de Baccharis dracunculifolia e Plantago lanceolata

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Improving mass rearing techniques for Cleruchoides noackae (Hymenoptera: Mymaridae)

The bronze bug, Thaumastocoris peregrinus, is an important pest affecting Eucalyptus plantations. The egg parasitoid Cleruchoides noackae was introduced in Brazil in 2012 for biological control of this pest. A mass rearing of C. noackae was established at EMBRAPA Florestas. This paper summarizes the main techniques developed to date in order to maximize the production of adult C. noackae. The use of eggs laid on towel paper strips increased the number of emerging parasitoids when compared to cutouts of Eucalyptus leaves. Host eggs 2 days old are preferred by C. noackae over 3- or 4-day-old ovipostures. Eggs can be stored at 5 ºC for 30 days after being parasitized without signifi cant effects on parasitoid emergence; such storage is a convenient strategy. The mean parasitoid emergence varies signifi cantly with the density of host eggs; an increase in the number of host eggs offered reduced the number of parasitoids that emerged. These improvements have played a signifi cant role in the production of C. noackae that has made possible mass release of C. noackae in Brazil and the establishment of natural populations of the parasitoid, as recently confi rmed.

Photosynthetic and anatomical responses of Eucalyptus grandis leaves to potassium and sodium supply in a field experiment

Although vast areas in tropical regions have weathered soils with low potassium (K) levels, little is known about the effects of K supply on the photosynthetic physiology of trees. This study assessed the effects of K and sodium (Na) supply on the diffusional and biochemical limitations to photosynthesis in Eucalyptus grandis leaves. A field experiment comparing treatments receiving K (+K) or Na (+Na) with a control treatment (C) was set up in a K-deficient soil. The net CO2 assimilation rates were twice as high in +K and 1.6 times higher in +Na than in the C as a result of lower stomatal and mesophyll resistance to CO2 diffusion and higher photosynthetic capacity. The starch content was higher and soluble sugar was lower in +K than in C and +Na, suggesting that K starvation disturbed carbon storage and transport. The specific leaf area, leaf thickness, parenchyma thickness, stomatal size and intercellular air spaces increased in +K and +Na compared to C. Nitrogen and chlorophyll concentrations were also higher in +K and +Na than in C. These results suggest a strong relationship between the K and Na supply to E. grandis trees and the functional and structural limitations to CO2 assimilation rates. © 2013 John Wiley & Sons Ltd.

Functional specialization of Eucalyptus fine roots: contrasting potential uptake rates for nitrogen, potassium and calcium tracers at varying soil depths

1. Little is known about the role of deep roots in the nutrition of forest trees and their ability to provide a safety-net service taking up nutrients leached from the topsoil. 2. To address this issue, we studied the potential uptake of N, K and Ca by Eucalyptus grandis trees (6 years of age - 25 m mean height), in Brazil, as a function of soil depth, texture and water content. We injected NO(3)(-)- (15)N, Rb(+) (analogue of K(+)) and Sr(2+) (analogue of Ca(2+)) tracers simultaneously in a solution through plastic tubes at 10, 50, 150 and 300 cm in depth in a sandy and a clayey Ferralsol soil. A complete randomized design was set up with three replicates of paired trees per injection depth and soil type. Recently expanded leaves were sampled at various times after tracer injection in the summer, and the experiment was repeated in the winter. Soil water contents were continuously monitored at the different depths in the two soils. 3. Determination of foliar Rb and Sr concentrations and (15)N atom % made it possible to estimate the relative uptake potential (RUP) of tracer injections from the four soil depths and the specific RUP (SRUP), defined as RUP, per unit of fine root length density in the corresponding soil layer. 4. The highest tracer uptake rates were found in the topsoil, but contrasting RUP distributions were observed for the three tracers. Whilst the RUP was higher for NO(3)(-)- (15)N than for Rb(+) and Sr(2+) in the upper 50 cm of soil, the highest SRUP values for Sr(2+) and Rb(+) were found at a depth of 300 cm in the sandy soil, as well as in the clayey soil when gravitational solutions reached that depth. 5. Our results suggest that the fine roots of E. grandis trees exhibit contrasting potential uptake rates with depth depending on the nutrient. This functional specialization of roots might contribute to the high growth rates of E. grandis trees, efficiently providing the large amounts of nutrients required throughout the development of these fast-growing plantations.

Chitinase and peroxidase activity in different stages of eucalypt leaves after inoculation with Puccinia psidii and acibenzolar-S-metil

Chitinase and peroxidase activity in different stages of eucalypt leaves after inoculation with Puccinia psidii and acibenzolar-S-metil To elucidate some biochemical processes during infection in the pathosystem Puccinia psidii x eucalyptus, the defense metabolism in different-stage leaves was compared between rust-resistant and susceptible clones, respectively. In addition, chitinase and peroxidase activities were assayed. Each treatment consisted of 4 replicates, in a completely randomized design: 2 clones, inoculated and not inoculated with P. psidii; sprayed with acibenzolar-S-methyl (ASM) and distilled water; and represented by the 1(st) leaf pair (size equivalent to 1/5 total leaf development), 2(nd) pair (2/5 total development), and 4(th) pair (4/5 total leaf length). Leaves were harvested in 4 periods: 0, 24, 72 and 96 hours after inoculation. Results indicated that ASM treatment or P. psidii action led to higher chitinase and peroxidase activity level but did not alter the expression of these activities in developed leaves (4(th) pair) during the experiment. Alterations in enzyme levels after inoculation were only observed in developing leaves (1(st) and 2(nd) pairs), which suggests that the response to infection was concomitant to chitinase and peroxidase synthesis. The highest increases in enzymatic activities were observed in resistant clones at 72 hours after inoculation and in susceptible ones previously treated with ASM and later inoculated with the pathogen.

Cultivo de Eucalyptus urograndis em atmosfera enriquecida com CO2: mudanças no proteoma cloroplastidial

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Studies on the eucalyptus leaf disease complex in Portugal

Native from south eastern Australia, Eucalyptus globulus is the main species in eucalypts plantations in Portugal. The most serious foliar disease in eucalypt plantations is linked to Mycosphaerella senso lato, which affects young trees in the juvenile phase foliage causing leaf necrosis. This disease results in reduced growth rate of the host and lower wood volume, thus causing significant productivity losses. The most common name for this disease was Mycosphaerella Leaf Disease that became inappropriate when most of the pathogens on eucalypts were re-distributed into several genera. The term "Eucalyptus Leaf Disease Complex" is now more appropriate. The overall aim of this thesis was to investigate the Eucalyptus Leaf Disease Complex in Portugal, focusing on species diversity, taxonomy and the role played by each species in the disease complex on Eucalyptus globulus. Literature on the Eucalyptus Leaf Disease Complex was reviewed and the species were distributed into several genera. A survey based on symptomatic leaves collected from several Eucalyptus globulus plantations and characterized by morphological and molecular tools provided an overview of species incidence and of the most frequent species in the disease complex. The present work reveals additional species of Mycosphaerella senso lato associated with eucalypt plantations in Portugal. Thus, five new records of Teratosphaeria and phylogenetically related species were added to the Iberian Peninsula, namely, Neodevriesia hilliana, for the first time on Myrtaceae; Quasiteratosphaeria mexicana, Teratosphaericola pseudoafricana, Teratosphaeria pluritubularis and Teratosphaeria lusitanica, a new species. Furthermore, new anamorphic structures were found and two new combinations were made. Regarding other genera, some species were observed for the first time, such as Cladosporium cladosporioides, Fusicladium eucalypti, Mycosphaerella madeirae, in the mainland. In addition to leave diseases, Teratosphaeria gauchensis was found causing a severe stem and trunk canker on Eucalyptus globulus. The aggressiveness of several species was compared to evaluate each species individually in the complex, permitting to distinguish different behaviours, from primary to secondary pathogens. Cladosporium cladosporioides, M. communis and M. lateralis, appeared to be more aggressive than Teratosphaeria nubilosa. In fact, contrary to the prevailing views on this disease complex, Teratosphaeria nubilosa is not the only species responsible for the disease, which clearly involves a complex of species acting together.

Boron mobility in eucalyptus clones

Understanding the magnitude of B mobility in eucalyptus may help to select clones that are more efficient for B use and to design new practices of B fertilization. This study consisted of five experiments with three eucalyptus clones (129, 57 and 58) where the response to and mobility of B were evaluated. Results indicated that clone 129 was less sensitive to B deficiency than clones 68 and 57, apparently due to its ability to translocate B previously absorbed via root systems to younger tissues when B in solution became limiting. Translocation also occurred when B was applied as boric acid only once to a single mature leaf, resulting in higher B concentration in roots, stems and younger leaves. The growth of B-deficient plants was also recovere by a single foliar application of B to a mature leaf. This mobility was greater, when foliar-applied B was supplied in complexed (boric acid + manitol) than in non-complexed form (boric acid alone). When the root system of clone 129 was split in two solution compartments, B supplied to one root compartment was translocated to the shoot and back to the roots in the other compartment, improving the B status and growth. Thus, it appears that B is relatively mobile in eucalyptus, especially in clone 129, and its higher mobility could be due to the presence of an organic compound such as manitol, able to complex B.