Eucalypt NADP-Dependent Isocitrate Dehydrogenase1 : cDNA Cloning and Expression in Ectomycorrhizae

NADP-dependent isocitrate dehydrogenase (NADP-ICDH) activity is increased in roots of Eucalyptus globulus subsp. bicostata ex Maiden Kirkp. during colonization by the ectomycorrhizal fungus Pisolithus tinctorius Coker and Couch. To investigate the regulation of the enzyme expression, a cDNA (EgIcdh) encoding the NADP-ICDH was isolated from a cDNA library of E. globulus-P. tinctorius ectomycorrhizae. The putative polypeptide sequence of EgIcdh showed a high amino acid similarity with plant NADP-ICDHs. Because the deduced EgICDH protein lacks an amino-terminal targeting sequence and shows highest similarity to plant cytosolic ICDHs, it probably represents a cytoplasmic isoform. RNA analysis showed that the steady-state level of EgIcdh transcripts was enhanced nearly 2-fold in ectomycorrhizal roots compared with nonmycorrhizal roots. Increased accumulation of NADP-ICDH transcripts occurred as early as 2 d after contact and likely led to the observed increased enzyme activity. Indirect immunofluorescence microscopy indicated that NADP-ICDH was preferentially accumulated in the epidermis and stele parenchyma of nonmycorrhizal and ectomycorrhizal lateral roots. The putative role of cytosolic NADP-ICDH in ectomycorrhizae is discussed.

Estabelecimento da composição volátil do nutriente FNI 210 usado pela Ceratitis capitata (Wiedemann, 1824) e dos extratos de diferentes espécies de plantas avaliação do potencial atrativo e repelente das plantas: avaliação do potencial atrativo e repelente das plantas

A mosca da fruta é uma das principais pragas agrícolas. Neste trabalho determinou-se a composição volátil do nutriente FNI 210 (proteína alimentar) e dos extratos de cinco plantas: Cedronella canariensis, Eucalyptus globulus, Laurus novocanariensis, Myrtus communis e Ruta chalepensis e avaliou-se o seu potencial atrativo e repelente em moscas adultas num olfatómetro em Y. A composição volátil do nutriente e dos extratos foi semelhante à encontrada por outros autores e apresentou compostos atrativos para a mosca da fruta. Nos bioensaios com o olfatómetro as moscas foram atraídas à proteína mas a percentagem média de respostas variou de acordo com o sexo, estado sexual, idade e número de indivíduos por grupo sendo mais alta aos 8 dias em grupos de 5. No geral, as fêmeas virgens responderam mais do que as não virgens e mais do que os machos virgens. O número de insetos que se dirigiram à proteína foi superior na primeira repetição nos primeiros 10 e 20 minutos. Contudo, em todos os bioensaios houve um número elevado de indivíduos não responderam. Nos bioensaios das plantas a resposta do mesmo grupo de 5 indivíduos com 8 dias foi testada três vezes no olfatómetro pela ordem seguinte: sem amostra, com proteína e com extrato de planta. Nos três casos as respostas dos adultos variaram de acordo com o sexo e estado sexual. As percentagens médias de respostas aos extratos foram superiores às obtidas nos ensaios sem amostra e menores que à proteína, á exceção do extrato de L. novocanariensis que apresentou um potencial atrativo superior ao da proteína nos machos virgens. Nos testes com o extrato, as respostas ao braço com amostra foram superiores ao braço sem amostra, à exceção das respostas das fêmeas não virgens ao extrato de R. chalepensis, o que sugere ser esta a única planta com potencial repelente.

Evolution of the landscape of Madeira Island: long-term vegetation dynamics

The aim of this thesis was to evaluate historical change of the landscape of Madeira Island and to assess spatial and temporal vegetation dynamics. In current research diverse “retrospective techniques”, such as landscape repeat photography, dendrochronology, and research of historical records were used. These, combined with vegetation relevés, aimed to gather information about landscape change, disturbance history, and vegetation successional patterns. It was found that landscape change, throughout 125 years, was higher in the last five decades manly driven by farming abandonment, building growth and exotic vegetation coverage increase. Pristine vegetation was greatly destroyed since early settlement and by the end of the nineteenth century native vegetation was highly devastated due to recurrent antropogenic disturbances. These actions also helped to block plant succession and to modify floristical assemblages, affecting as well as species richness. In places with less hemeroby, although significant growth of vegetation of lower seral stages was detected, the vegetation of most mature stages headed towards unbalance between recovery and loss, being also very vulnerable to exotic species encroachment. Recovery by native vegetation also occurred in areas formerly occupied by exotic plants and agriculture but it was almost negligible. Vegetation recovery followed the successional model currently proposed, attesting the model itself. Yet, succession was slower than espected, due to lack of favourable conditions and to recurrent disturbances. Probable tempus of each seral stage was obtained by growth rates of woody taxa estimated through dendrochronology. The exotic trees which were the dominant trees in the past (Castanea sativa and Pinus pinaster) almost vanished. Eucalyptus globulus, the current main tree of the exotic forest is being replaced by other cover types as Acacia mearnsii. The latter, along with Arundo donax, Cytisus scoparius and Pittosporum undulatum are currently the exotic species with higher invasive behaviour. However, many other exotic species have also proved to be highly pervasive and came together with the ones referred above to prevent native vegetation regeneration, to diminish biological diversity, and to block early successional phases delaying native forest recovery.

The Evolving Nature of Small-Scale Forestry in Australia

Two forms of small-scale forestry are developing in Australia, each with different impacts on rural communities. One is based on growing short-rotation Eucalyptus globulus (blue gum) for pulp and the other on production of higher-value products from longer-rotation native hard-woods. Several impediments exist to further development of small-scale forestry, including the lack of a small-scale forestry culture, concerns over harvest rights, lack of market development, the long wait for returns, and satisfaction with current land uses. Nevertheless, the rapid increase in farm woodlot establishment in the past five years has paralleled the strong increase in the private industrial plantation estate. As markets develop and hindrances are overcome, landholders not previously interested in small-scale forestry may consider ita worthwhile land use.

Modelling predicts positive and negative interactions between three Australian tropical tree species in monoculture and binary mixture

Computer modelling promises to. be an important tool for analysing and predicting interactions between trees within mixed species forest plantations. This study explored the use of an individual-based mechanistic model as a predictive tool for designing mixed species plantations of Australian tropical trees. The 'spatially explicit individually based-forest simulator' (SeXI-FS) modelling system was used to describe the spatial interaction of individual tree crowns within a binary mixed-species experiment. The three-dimensional model was developed and verified with field data from three forest tree species grown in tropical Australia. The model predicted the interactions within monocultures and binary mixtures of Flindersia brayleyana, Eucalyptus pellita and Elaeocarpus grandis, accounting for an average of 42% of the growth variation exhibited by species in different treatments. The model requires only structural dimensions and shade tolerance as species parameters. By modelling interactions in existing tree mixtures, the model predicted both increases and reductions in the growth of mixtures (up to +/- 50% of stem volume at 7 years) compared to monocultures. This modelling approach may be useful for designing mixed tree plantations. (c) 2006 Published by Elsevier B.V.

Planted forests and biodiversity

Expansion of planted forests and intensification of their management has raised concerns among forest managers and the public over the implications of these trends for sustainable production and conservation of forest biological diversity. We review the current state of knowledge on the impacts of plantation forestry on genetic and species diversity at different spatial scales and discuss the economic and ecological implications of biodiversity management within plantation stands and landscapes. Managing plantations to produce goods such as timber while also enhancing ecological services such as biodiversity involves tradeoffs, which can be made only with a clear understanding of the ecological context of plantations in the broader landscape and agreement among stakeholders on the desired balance of goods and ecological services from plantations.

Environmental impacts of freshwater use and suspended solids in life cycle assessment

Perturbation of natural ecosystems, namely by increasing freshwater use and its degradative use, as well as topsoil erosion by water of land-use production systems, have been emerging as topics of high environmental concern. Freshwater use has become a focus of attention in the last few years for all stakeholders involved in the production of goods, mainly agro-industrial and forest-based products, which are freshwater-intensive consumers, requiring large inputs of green and blue water. This thesis presents a global review on the available Water Footprint Assessment and Life Cycle Assessment (LCA)-based methods for measuring and assessing the environmental relevance of freshwater resources use, based on a life cycle perspective. Using some of the available midpoint LCA-based methods, the freshwater use-related impacts of a Portuguese wine (white ‘vinho verde’) were assessed. However, the relevance of environmental green water has been neglected because of the absence of a comprehensive impact assessment method associated with green water flows. To overcome this constraint, this thesis helps to improve and enhance the LCA-based methods by providing a midpoint and spatially explicit Life Cycle Impact Assessment (LCIA) method for assessing impacts on terrestrial green water flow and addressing reductions in surface blue water production caused by reductions in surface runoff due to land-use production systems. The applicability of the proposed method is illustrated by a case study on Eucalyptus globulus conducted in Portugal, as the growth of short rotation forestry is largely dependent on local precipitation. Topsoil erosion by water has been characterised as one of the most upsetting problems for rivers. Because of this, this thesis also focuses on the ecosystem impacts caused by suspended solids (SS) from topsoil erosion that reach freshwater systems. A framework to conduct a spatially distributed SS delivery to freshwater streams and a fate and effect LCIA method to derive site-specific characterisation factors (CFs) for endpoint damage on aquatic ecosystem diversity, namely on algae, macrophyte, and macroinvertebrates organisms, were developed. The applicability of this framework, combined with the derived site-specific CFs, is shown by conducting a case study on E. globulus stands located in Portugal as an example of a land use based system. A spatially explicit LCA assessment was shown to be necessary, since the impacts associated with both green water flows and SS vary greatly as a function of spatial location.

Extração de ácidos triterpénicos a partir da casca de eucalipto utilizando líquidos iónicos

Os recursos renováveis têm sido um forte alvo de investigação científica nos últimos anos, onde o aproveitamento de biomassa e seus resíduos para a obtenção de compostos de valor acrescentado, combustíveis e energia têm sido abordados no conceito de biorrefinaria integrada. As indústrias de papel geram quantidades significativas de resíduos, nomeadamente a casca de eucalipto que é atualmente queimada para a geração de energia. De forma a valorizar este resíduo, a presente dissertação teve como objetivo extrair compostos triterpénicos, a partir casca externa de Eucalyptus globulus, utilizando solventes de extração alternativos - soluções aquosas de líquidos iónicos (LIs) – para substituir os solventes orgânicos actualmente utilizados. Os ácidos triterpénicos apresentam um elevado interesse na indústria cosmética, farmacêutica e alimentar graças às suas propriedades antiinflamatórias, antitumurais, entre outras. Primeiramente, caracterizou-se a casca externa de Eucalyptus globulus, e posteriormente procedeu-se ao estudo de solubilidade de ácido ursólico (AU, utilizado como molécula modelo) a 25 ºC em soluções aquosas de LIs e surfactantes de modo a selecionar os solventes mais eficientes para a extração. Deste trabalho conclui-se que a capacidade surfactante das soluções aquosas de LIs, particularmente [C4C1im][C8H17SO4], [C16C1im]Cl e [C14C1im]Cl, desempenham um papel fundamental para a solubilização de AU em água, podendo aumentar quase 16000 vezes a sua solubilidade, e permitiu recuperar cerca de 89% deste composto com simples adição de água como anti-solvente. Por fim, compararam-se as quantidades de ácidos triterpénicos extraídas a partir da casca de eucalipto com soluções aquosas de [C14C1im]Cl, metanol e com extração em soxhlet com diclorometano.

Production of bioplastics from hydrolysates of paper industry by Haloferax mediterranei

The biorefinery concept has attracted much attention over the last decade due to increasing concerns about the use of fossil resources. In this context emerged the use of bioplastics, namely polyhydroxyalkanoates (PHA). PHA are biocompatible and biodegradable plastics that can be obtained from renewable raw materials and can constitute an alternative solution to conventional plastics. In this work, hydrolysed cellulose pulp, coming from Eucalyptus globulus wood cooking, was used as substrate to the PHA-storing bacteria Haloferax mediterranei. The hydrolysed pulp is rich in simple sugars, mainly glucose (81.96 g.L-1) and xylose (20.90 g.L-1). Tests were made in defined medium with glucose and xylose and in hydrolysate supplemented with salts and yeast extract. Different concentrations of glucose were tested, namely 10, 15, 20, 30 and 40 g.L-1. The best accumulation results (27.1 % of PHA) were obtained in hydrolysate medium with 10 g.L-1. Using this concentration, assays were performed in fed-batch and sequencing batch reactor conditions in order to determine the best feeding strategy. The strategy that led to the best results was fed-batch assay with 24.7 % of PHA. An assay without sterile conditions was performed, in which was obtained the same growth than in sterilization test. Finally it was performed an assay in a bioreactor and a fast growth (0.14 h-1) with high glucose and xylose consumption rates (0.368 g.L-1.h-1 and 0.0947 g.L-1.h-1, respectively) were obtained. However 1.50 g.L-1 of PHA, corresponding to 16.1 % (92.52 % of 3HB and 3HV of 7.48 %) of % PHA were observed. The polymer was further characterized by DSC with a glass transition temperature of -6.07 °C, a melting temperature of 156.3 °C and a melting enthalpy of 63.07 J.g-1, values that are in accordance with the literature. This work recognizes for the first time the suitability of the pulp paper hydrolysate as a substrate for PHA production by H. mediterranei.

Contribution of plant phytochemicals to organic homeostasis: focus on the aging process and opportunistic infections

Aging process is conceived as a normal stage during human life cycle, but it is also considered a hot topic among scientists and medical community. Alarming rates of premature aging and oxidative stress-related diseases have increasingly affect human individuals. Stress, pollution and exposition to chemical substances are considered the main triggering factors for those conditions; in addition, they also suppress the immune system and, therefore, improve organic vulnerability and occurrence of opportunistic infections [I]. Apart from the associated morbidity and mortality, the increasing rates of antimicrobial resistance improve the severity of the clinical conditions [2]. Botanical preparations possess a multitude of bioactive properties, namely acting as antimicrobials, antioxidants, and homeostasis modulators. Thus, upcoming alternatives, mainly based in plant phytochemicals, are necessary to improve the wellbeing as also life expectancy of individuals. The present study aims to evaluate and to compare both antioxidant and antimicrobial properties of plant extracts rich in phenolic compounds. Among the tested plants, Glycyrrhiza glabra L. (licorice) evidenced the most pronounced free radicals scavenging and antimicrobial effects, followed by Salvia officina/is L. (sage), Thymus vulgaris L. (thyme) and Origanum vulgare L. (oregano). Eucalyptus globulus Labill. (blue gum) and Juglans regia L. (walnut) also showed a high effect, while Pterospartum tridentatum (L.) Willk. (carqueja) and Rubus ulmifolius Schott (elm leaf blackberry) displayed moderate effects, and lastly, Tabebuia impetigirwsa (Mart. ex DC) Standley (pau d'arco), Foeniculum vulgare Miller (fennel), Rosa canina L. (rose hips) and Matricaria recutita L. (chamomile) gave only slight effects. In general, the most pronounced bioactivities were observed in the plant preparations (infusion>decoction>hydromethanolic extract) with higher levels of phenolic compounds (both flavonoids and phenolic acids). The observed synergisms between the phenolic compounds present in the extracts highlight the use of phytochemicals as future health promoters. However, further studies are necessary to understand the effective mode of action of individual phenolic constituents as also the existence of polyvalence relationships between them.

Estudo da bioecologia de Rhombacus eucalypti Ghosh & Chakrabarti, um ácaro eriofídeo do eucalipto em Portugal

Mestrado em Engenharia Agronómica - Instituto Superior de Agronomia - UL

Eucalyptus sp seedling response to potassium fertilization and soil water

A considerable portion of Brazil's commercial eucalypt plantations is located in areas Subjected to periods of water deficit and grown in soils with low natural fertility, particularly poor In potassium. Potassium is influential in controlling water relations of plants. The objective of this study was to verify the influence of potassium fertilization and soil water potential (psi(w)) oil the dry matter production and oil water relations Of eucalypt seedlings grown under greenhouse conditions. the experimental units were arranged in 4x4x2 randomized blocks factorial design, as follow: four species of Eucalyptus (Eucalyptus grandis, Eucalyptus urophylla, Eucalyptus camaldulensis and hybrid Eucalyptus grandis x Eucalyptus urophylla), four dosages of K (0, 50, 100 and 200 mg dm(-3)) and two soil water potentials (-0.01 M Pa and -0.1 M Pa). Plastic containers with 15 cm diameter and 18 cm height, with Styrofoam base, containing 3.0 dm(3) of soil and two plants per container were used. Soil water potential was kept at -0.01 MPa for 40 days after seeding. Afterward, the experimental units were divided into two groups: in one group the potential was kept at 0.01 MPa, and in the other one, at -0.10 MPa. Sol I water potential was control led gravimetrically twice a day with water replacement until the desired potential was reestablished. A week before harvesting, the leaf water potential (psi), the photosynthetic rate (A), the stomatal conductance (gs) and the transpiration rate were evaluated. The last week before harvesting, the mass of the containers was recorded daily before watering to determine the consumption of water by the plants. After harvesting, total dry matter and leaf area were evaluated. the data were Submitted to analysis of variance, to Tukey's tests and regression analyses. The application of K influenced A, gs and the transpiration rate. Plants deficient in K showed lower A and higher Us and transpiration rates. There were no statistical differences in A, gs and transpiration rates ill plants with and Without water deficit. The addition of K reduced the consumption of water per unit of leaf area and, in general, plants submitted to water deficit presented a lower consumption of water.

Efeito da aplicação de fertilização nitrogenada e lodo de esgoto nas características da madeira juvenil de árvores de Eucalyptus grandis

This work evaluated the wood anatomical and physical characteristics of 24 months-old Eucalyptus grandis trees, planted in 3x2m spacing and fertilized with nitrogen (6, 12, 18 month old) and sewage sludge (planting, 8 month old). For each treatment 10 eucalypts trees were cut according to the distribution of basal area. Wood samples were collected in different percentages of the total height to analyze the anatomical (vessels and fibers) and physical (wood density) characteristics. The results showed that the wood apparent density and wood basic density of the eucalypt trees in the nitrogen and sewage sludge were larger in comparison to the control. Radial profiles of wood apparent density, were similar in the three treatments, presenting the exected characteristics of juvenile wood of 24 months-old eucalypt trees. Fiber and vessel dimensions were not affected by fertilization.

Can Eucalyptus invade native forest fragments close to commercial stands?

Some Eucalyptus species are widely used as a plantation crop in tropical and subtropical regions. One reason for this is the diversity of end uses, but the main reason is the high level of wood production obtained from commercial plantings. With the advancement of biotechnology it will be possible to expand the geographical area in which eucalypts can be used as commercial plantation crops, especially in regions with current climatic restrictions. Despite the popularity of eucalypts and their increasing range, questions still exist, in both traditional planting areas and in the new regions: Can eucalypts invade areas of native vegetation, causing damage to natural ecosystems biodiversity? The objective of this study it was to assess whether eucalypts can invade native vegetation fragments in proximity to commercial stands, and what factors promote this invasive growth. Thus, three experiments were established in forest fragments located in three different regions of Brazil. Each experiment was composed of 40 plots (1 m(2) each one), 20 plots located at the border between the forest fragment and eucalypts plantation, and 20 plots in the interior of the forest fragments. In each experimental site, the plots were paired by two soil exposure conditions, 10 plots in natural conditions and 10 plots with soil exposure (no plant and no litter). During the rainy season, 2 g of eucalypts seeds were sown in each plot, including Eucalyptus grandis or a hybrid of E. urophylla x E. grandis, the most common commercial eucalypt species planted in the three region. At 15, 30, 45, 90, 180, 270 and 360 days after sowing, we assessed the number of seedlings of eucalypts and the number of seedlings of native species resulting from natural regeneration. Fifteen days after sowing, the greatest number of eucalypts seedlings (37 m(-2)) was observed in the plots with lower luminosity and exposed soil. Also, for native species, it was observed that exposed soil improved natural germination reaching the highest number of 163 seedlings per square meter. Site and soil exposure were the factors that have the greatest influence on seed germination of both eucalypt and native species. However, 270 days after sowing, eucalypt seedlings were not observed at any of the three experimental sites. The result shows the inability of eucalypts to adapt to condition outside of their natural range. However, native species demonstrated their strong capacity for natural regeneration in forest fragments under the same conditions where eucalypts were seeded. (C) 2011 Elsevier B.V. All rights reserved.

Inheritance of resistance to Puccinia psidii G. Winter in a eucalyptus interspecific hybrid progeny evaluated under conditions of natural infection

Inheritance of resistance to Puccinia psidii G. Winter in a eucalyptus interspecific hybrid progeny evaluated under conditions of natural infection Rust caused by the fungus Puccinia psidii is currently the most important disease of eucalyptus. It is widely disseminated in Brazil, and causes serious damage in nurseries and plantation areas. The identification of resistant germplasm along with knowledge of the genetic basis of resistance heredity are the first requirements for the success of breeding programs aiming to develop resistant varieties. Earlier studies carried out under controlled conditions suggested a monogenic control as well as the participation of at least two genes promoting resistance to rust. The goal of this study was to evaluate the resistance to P. psidii under field conditions in fourteen progenies from controlled crosses and self-crosses among four hybrid clones of Eucalyptus grandis Hill ex Maiden x Eucalyptus urophylla ST Blake that contrast for resistance to the fungus. Results indicated that resistance could be explained by one locus with main effects and at least three different alleles. However, loci with minor effects may influence the resistance, since variation on severity classes was observed. Differences in segregation of resistance between reciprocal crosses were not observed, indicating absence of cytoplasmic effects.