Phenolic compounds in Coleostephus myconis (L.) Rchb.f.: characterization by HPLC-DAD-ESI/MS and phenology effects

The Asteraceae family is spread worldwide. In Portugal, there are more than 300 species, standing out as one of the botanical families with largest representation in the Portuguese flora. Coleostephus myconis (L.) Rchb.f. is a scarcely studied Asteraceae species, characterized as having ruderal growth and persistence in abandoned soils (an expanding problem due to the desertification phenomena in rural areas). In this work, the flowers of C. myconis were collected in three different flowering stages (i: flower bud; ii: flower in anthesis; iii: senescent flower) from the Northwestern area of the Portuguese territory. Powdered samples (1 g) were extracted twice with ethanol:water 50:50 (v/v). After removing solvents, the combined extracts were re-dissolved, filtered through 0.22-μm disposable LC filter disks and analyzed by high performance liquid chromatography coupled to a diode array detector and electrospray ionization-mass spectrometry (HPLC-DAD/ESI-MS). The phenolic compounds were characterized according to their UV and mass spectra, and retention times. For the quantitative analysis, calibration curves of standard compounds were used. According to the UV spectra (λmax = 314-330 nm) and pseudomolecular ions ([M-H]-) at m/z 353 and 515, all producing an m/z 191 ion, four compounds derived from quinic acid were detected: 3-O-caffeoylquinic acid (Figure 1A), 5-O-caffeoylquinic acid (Figure 1B), 3,5-O-dicaffeoylquinic acid (Figure 1C) and 4,5-O-dicaffeoylquinic acid (Figure 1D), as also supported by the literature [1,2]. A fifth phenolic acid was identified as protocatechuic acid. The detected flavonoid were quercetin-O-glucuronide, quercetin-3-Oglucoside, myricetin-O-methyl-hexoside and a second glycosylated myricetin (not possible to identify completely). Some statistically significant changes were detected among the different assayed flowering stages; nevertheless, 3,5-O-dicaffeoylquinic acid was the major compound, independently of the phenologic stage. According to the previous results, C. myconis might be considered as a potential natural source of these valuable bioactive compounds, especially considering the high botanical representativeness of this plant and its inexpensiveness.

Propagação de Camellia sinensis: efeito do genótipo, estaca, substrato, recipiente e ácido indolbutírico

Tem sido relatado que as estacas de Camellia sinensis possuem baixa capacidade de emitir raízes, motivando assim a realização de estudos básicos para otimização do processo de propagação por estacas. Assim sendo, o presente trabalho objetivou quantificar o potencial rizogênico de diferentes genótipos e o efeito da posição da estaca no ramo e incisão na base, do substrato, tamanho do recipiente e ácido indolbutírico no enraizamento de estacas semi-lenhosas dessa espécie. Para tal, foram coletados ramos dos genótipos IAC 259, F15 e Comum, em Pariquera-Açu-SP, no inverno de 2010. em seguida, preparadas as estacas, contendo uma gema e uma folha, foram mantidas em viveiro com 70% de sombreamento. Estacas da posição basal e mediana dos ramos são as mais adequadas para estaquia devido a menor mortalidade e maior enraizamento. A injúria na base da estaca não afeta a mortalidade e o enraizamento das estacas, porém induz à formação de calo. Também não houve diferenças na mortalidade e no enraizamento das estacas quando as mesmas foram mantidas em recipiente de 50, 90 e 120 cm³. Comparado com vermiculita, areia e casca de arroz carbonizada, o solo foi o melhor substrato para estaquia, que na presença do ferimento, juntamente com o tratamento das estacas com 10 g L-1 de AIB promoveu a maior porcentagem de enraizamento. Todavia, ainda nessa condição a mortalidade média das estacas foi de 42%. O potencial de enraizamento do genótipo Comum foi superior ao do IAC 259 e F15.

Bioorthogonal Noncanonical Amino Acid Tagging for Selective Analysis of the Pseudomonas aeruginosa Proteome

In natural environments, bacterial physiology is frequently characterized by slow metabolic rates and complex cellular heterogeneities. The opportunistic pathogen Pseudomonas aeruginosa provides one such example; P. aeruginosa forms untreatable chronic biofilm infections of the cystic fibrosis lung, where oxygen limitation can lead to states of metabolic dormancy. To better understand the biology of these states, in vitro experiments must be adapted to better recapitulate natural settings. However, low rates of protein turnover and cellular or phenotypic complexity make these systems difficult to study using established methods. Here we adapt the bioorthogonal noncanonical amino acid tagging (BONCAT) method for time- and cell-selective proteomic analysis to the study of P. aeruginosa. Analysis of proteins synthesized in an anoxic dormancy state led to the discovery of a new type of transcriptional regulator which we designated SutA. We performed detailed analyses of SutA’s role in transcription under slow growth states and we elucidated the structural basis for its regulatory behavior. Additionally, we used cell-selective targeting of BONCAT labeling to measure the dynamic proteomic response of an antibiotic-tolerant biofilm subpopulation. Overall this work shows the utility of selective proteomics as applied to bacterial physiology and describes the broad biological insight obtained from that application.

Influence of mother plant and seed coat colour on germination success of Retama sphaerocarpa L (Boissier)

Mestrado em Engenharia Florestal e dos Recursos Naturais - Instituto Superior de Agronomia - UL

Seven Ulmus minor clones tolerant to Ophiostoma novo-ulmi registered as forest reproductive material in Spain

The Spanish elm programme began in 1986 in response to the devastating impact of Dutch elm disease on natural elm stands and urban trees. Its main objectives were to conserve remaining genetic resources and select and breed tolerant native elm genotypes. After 27 years of work conducting susceptibility trials on thousands of elm genotypes, the first seven tolerant Ulmus minor trees are now being registered by the Spanish Environmental Administration. This paper presents the results of the susceptibility tests on these clones and their distinctive genetic, morphological and phenological features. In all susceptibility trials the commercial ?Sapporo Autumn Gold? clone, which is highly tolerant to O. novo-ulmi, was used as a control. The registered clones were named ?Ademuz?, ?Dehesa de la Villa?, ?Majadahonda?, ?Toledo?, ?Dehesa de Amaniel?, ?Retiro? and ?Fuente Umbría?. The most tolerant clone was ?Dehesa de Amaniel?, as its wilting values were below 5% during the two consecutive inoculation trials performed in Madrid. ?Fuente Umbría?, tested over four consecutive years in Guadalajara and Palencia, was the Spanish clone with the most reliable tolerance level to O. novo-ulmi. The ?Ademuz? and ?Majadahonda? clones had the highest ornamental scores and are promising trees for use in urban environments and tree breeding for ornamental quality. These two genotypes showed a later bud burst phenology than the other U. minor clones, demonstrating suitability to areas with late frost events. The Spanish programme aims to substantially increase the range of tolerant native elms through new selections and crossings to gain a better understanding of the genetic basis of resistance.

Ammonium uptake, transport and nitrogen economy in forest trees

AMMONIUM UPTAKE, TRANSPORT AND NITROGEN ECONOMY IN FOREST TREES Francisco M. Cánovas, Concepción Avila, Fernando N. de la Torre, Rafael A. Cañas, Belén Pascual, Vanessa Castro- Rodríguez, Jorge El-Azaz Departamento de Biología Molecular y Bioquímica, Facultad de Ciencias, Universidad de Málaga, Spain. Email: canovas@uma.es Forests ecosystems play a fundamental role in the regulation of global carbon fixation and preservation of biodiversity. Forest trees are also of great economic value because they provide a wide range of products of commercial interest, including wood, pulp, biomass and important secondary metabolites. The productivity of most forest ecosystems is limited by low nitrogen availability and woody perennials have developed adaptation mechanisms, such as ectomycorrhizal associations, to increase the efficiency of N acquisition and metabolic assimilation. The efficient acquisition, assimilation and economy of nitrogen are of special importance in trees that must cope with seasonal periods of growth and dormancy over many years. In fact, the ability to accumulate nitrogen reserves and to recycle N is crucial to determine the growth and production of forest biomass. Ammonium is the predominant form of inorganic nitrogen in the soil of temperate forests and many research efforts are addressed to study the regulation of ammonium acquisition, assimilation and internal recycling for the biosynthesis of amino acids, particularly those relevant for nitrogen storage. In our laboratory, we are interested in studying nitrogen metabolism and its regulation in maritime pine (Pinus pinaster L. Aiton), a conifer species of great ecological and economic importance in Europe and for which whole-transcriptome resources are available. The metabolism of phenylalanine plays a central role in the channeling of carbon from photosynthesis to the biosynthesis of phenylpropanoids and the regulation of this pathway is of broad significance for nitrogen economy of maritime pine. We are currently exploring the molecular properties and regulation of genes involved in the biosynthesis and metabolic fates of phenylalanine in maritime pine. An overview of this research programme will be presented and discussed. Research supported by Spanish Ministry of Economy and Competitiveness and Junta de Andalucía (Grants BIO2015-69285-R, BIO2012-0474 and research group BIO-114).

Germinação de sementes e otimização de técnicas de micropropagação de umbuzeiro (spondias tuberosa, arr.) anacardiaceae

The brazilian-plum (Spondias tuberosa, His) is a tropical fruit tree that has been consolidated in the market for agribusiness processing, due to its characteristic flavor of fruit. Accordingly, studies to optimize the propagation of plants are necessary for production of seedlings with agronomic and quality assurance measures. This study aimed at determining the efficient techniques for uniform seed germination, as brazilian-plum seed present mechanical dormancy, and establish optimal culture media for multiplication of shoots from the in vitro micropropagation. Firstly, in a greenhouse at the Universidade Federal do Rio Grande do Norte, was evaluated the influence of different methods of breaking dormancy in the emergence of seedlings of brazilian-plum and speed of germination (IVG) of seeds. After 60 days of cultivation, it was found that splay in the distal portion of the seed was the best treatment, with rates of 85.33% in germinability and 3.415 of IVG, compared with the treatment of seed-soaking in water for 12h + humus and the control group. Subsequently, new sources of seedling explants were obtained in studies of tissue culture. Laboratory of Plant Biotechnology that the university, was used stem apex, nodal segments and internodes in search of decontamination with various concentrations of calcium hypochlorite [Ca(OCl)2] and micropropagation, inoculating them in half WPM (1980) with various concentrations of 6-benzylaminopurine (BAP). We used 10 sample units with three replications for different concentrations of [Ca(OCl)2], BAP and explants type. After thirty days, which was observed for the control of contamination, during the establishment in vitro, concentrations of [Ca(OCl)2] between 0.5% and 2.0% were effective in combating exogenous contamination of the apex. In nodal segments and internodes, concentrations of [Ca(OCl)2] between 1.0% and 2.0% and 1.5% and 2.0% were respectively, sufficient to reduce the percentage of losses in these infestations explants. For micropropagation, the culture medium supplemented with 0.1 mg.L-1 BAP promotes better development of multiple shoots per explants from nodal segment. However, success does not get to shoot training in internodal segment

Aedes aegypti Pharate First Instar Aseasonal Quiescence Cues Anticipatory Plasticity with Implications for Urban Vector Ecology and Control

The eggs of the dengue fever vector Aedes aegypti possess the ability to undergo an extended quiescence period hosting a fully developed first instar larvae within its chorion. As a result of this life history stage, pharate larvae can withstand months of dormancy inside the egg where they depend on stored reserves of maternal origin. This adaptation known as pharate first instar quiescence, allows A. aegypti to cope with fluctuations in water availability. An examination of this fundamental adaptation has shown that there are trade-offs associated with it. Aedes aegypti mosquitoes are frequently associated with urban habitats that may contain metal pollution. My research has demonstrated that the duration of this quiescence and the extent of nutritional depletion associated with it affects the physiology and survival of larvae that hatch in a suboptimal habitat; nutrient reserves decrease during pharate first instar quiescence and alter subsequent larval and adult fitness. The duration of quiescence compromises metal tolerance physiology and is coupled to a decrease in metallothionein mRNA levels. My findings also indicate that even low levels of environmentally relevant larval metal stress alter the parameters that determine vector capacity. My research has also demonstrated that extended pharate first instar quiescence can elicit a plastic response resulting in an adult phenotype distinct from adults reared from short quiescence eggs. Extended pharate first instar quiescence affects the performance and reproductive fitness of the adult female mosquito as well as the nutritional status of its progeny via maternal effects in an adaptive manner, i.e., anticipatory phenotypic plasticity results as a consequence of the duration of pharate first instar quiescence and alternative phenotypes may exist for this mosquito with quiescence serving as a cue possibly signaling the environmental conditions that follow a dry period. M findings may explain, in part, A. aegypti’s success as a vector and its geographic distribution and have implications for its vector capacity and control.

Environmental factors correlated with the metabolite profile of Vitis vinifera cv. Pinot Noir berry skins along a European latitudinal gradient

Mature berries of Pinot Noir grapevines were sampled across a latitudinal gradient in Europe, from southern Spain to central Germany. Our aim was to study the influence of latitude-dependent environmental factors on the metabolite composition (mainly phenolic compounds) of berry skins. Solar radiation variables were positively correlated with flavonols and flavanonols and, to a lesser extent, with stilbenes and cinnamic acids. The daily means of global and erythematic UV solar radiation over long periods (bud break-veraison, bud break-harvest, and veraison-harvest), and the doses and daily means in shorter development periods (5–10 days before veraison and harvest) were the variables best correlated with the phenolic profile. The ratio between trihydroxylated and monohydroxylated flavonols, which was positively correlated with antioxidant capacity, was the berry skin variable best correlated with those radiation variables. Total flavanols and total anthocyanins did not show any correlation with radiation variables. Air temperature, degree days, rainfall, and aridity indices showed fewer correlations with metabolite contents than radiation. Moreover, the latter correlations were restricted to the period veraison-harvest, where radiation, temperature, and water availability variables were correlated, making it difficult to separate the possible individual effects of each type of variable. The data show that managing environmental factors, in particular global and UV radiation, through cultural practices during specific development periods, can be useful to promote the synthesis of valuable nutraceuticals and metabolites that influence wine quality.

Defeitos de desenvolvimento do esmalte e respetivo tratamento

As estruturas dentárias são revestidas pelo esmalte dentário. O esmalte é um tecido de alta dureza, avascular e predominantemente branco. No entanto, distingue-se dos outros tecidos mineralizados do corpo pela sua incapacidade de remodelação. Devido a esse facto qualquer alteração que ocorra, quer ao longo da vida, quer no seu desenvolvimento fica, permanentemente, registada (Seow, 1997). Procurou-se nesta monografia aprofundar os conhecimentos sobre os mais comuns defeitos de desenvolvimento do esmalte existentes, assim como o respetivo tratamento. Para a realização desta monografia foram utilizados os seguintes motores de busca B-on, PubMed, Science Direct e Sci-elo, para a realização da pesquisa de informação, aplicando-se um critério de seleção temporal dos últimos 10 anos. As palavras-chaves e combinações de palavras utilizadas nos motores de busca referidos para a realização da pesquisa foram “Enamel”, “Enamel Development”, “Enamel Defects”, “Amelogenisis Imperfecta”, “Hypoplasia”. Dos 300 artigos encontrados nesta pesquisa, foram selecionados 68. O desenvolvimento dos tecidos dentários é um processo complexo conhecido por odontogénese, podendo ser simplisticamente dividido em três fases Fase de Botão, Fase de Capuz e por último a Fase de Campânula (Thesleff et al.,2009) Existem inúmeros defeitos de desenvolvimento do esmalte registados na literatura, não sendo mesmo possível em muitos casos enquadrar indubitavelmente o referido defeito numa categoria, ou até atribuir-lhe uma designação (Seow, 1997). Optou-se pela sua relevância e epidemiologia abordar nesta monografia os seguintes defeitos: Defeitos de desenvolvimento do esmalte; Opacidades; Opacidade difusa; Hipoplasia; Amelogenese imperfeita e todas as suas categorias; Fluorose e manchas por tetraciclinas assim como os seus respectivos tratamentos. Os defeitos de desenvolvimento de esmalte apresentam diversas características próprias e outras semelhantes entre si, verificando-se assim diversas possibilidades de tratamentos a realizar, uns mais invasivos e outros menos, que vão desde microabrasões na superfície do esmalte, à colocação de cerâmicas, dependendo sempre da preferência do paciente e do seu poder socioeconómico (Azevedo DT et al., 2011). Conclui-se que apesar de todos os problemas que acarretam quer a nível estético quer a nível funcional para os indivíduos nos quais não existe uma grande gravidade das lesões esses casos podem ser resolvidos por um Médico Dentista generalista desde que este tenha o conhecimento adequado dos protocolos de atuação.

Bunch yield of interspecific hybrids of American oil palm with oil palm in the juvenile phase.

To identify superior genotypes, 42 progenies of interspecific hybrids between American oil palm and oil palm were evaluated in an area with incidence of bud rot. The following variables were assessed: total bunch yield (TBY), number of bunches (NB) and mean bunch weight (MBW) from the third to the sixth year of cultivation and estimates of genetic parameters obtained by REML/BLUP procedures.

Starch, growth, and development in cuttings of rootstock "Paulsen 1103" from plants infested with ground pearl.

ABSTRACT The ground pearl, Eurhizococcus brasiliensis, is considered an important pest of vineyards in southern Brazil, with affected plants exhibiting leaf chlorosis, reduction in vigor, fading, and death. This study evaluated the quality of hardwood cuttings produced from plants infected (I) and not infected (NI) by ground pearl. ?Paulsen 1103? (Vitis berlandieri × Vitis rupestris) plants were grown for 29 months in brick-built raised beds either infested or not infested by ground pearl; then, 12 one-year-old branches with a maximum of 12 buds each were cut from each plant, subdivided into three portions (4 buds cutting-1), and subjected to destructive and nondestructive testing. Destructive testing comprised determining fresh and dry weight, length, internode diameters, and percentage of starch. Nondestructive testing comprised assessing the potential for bud sprouting and shoot development. Each mother plant in the I and NI beds was considered a replicate, with a total of 360 cuttings per treatment. It was observed that cuttings from infected plants had significantly lower (P<0.05) internode diameter, length, and fresh and dry weight than those of the uninfected plants. The percentage of starch content of the cuttings did not differ significantly. All cuttings showed the same percentage (100%) of bud breaking and no changes in growth and development of seedlings regardless of source. Given these results, it was concluded that vines of ?Paulsen 1103? infested with ground pearl produce smaller cuttings than those of uninfected plants but with no reduction in bud break percentage or seedling development. Key words: Margarodidae, Vitaceae, insect?plant interaction, carbohydrates. RESUMO A pérola-da-terra, Eurhizococcus brasiliensis, tem sido considerada uma importante praga dos vinhedos no sul do Brasil, sendo que as plantas atacadas manifestam clorose foliar, redução no vigor, definhamento e morte. Este trabalho teve por objetivo avaliar a qualidade de estacas lenhosas produzidas a partir do contraste de videiras infestadas (I) e não infestadas (NI) por pérola-da-terra. Após 29 meses de cultivo em canteiros de alvenaria, em presença ou ausência de pérola-da-terra, cada planta da variedade ?Paulsen 1103? (Vitis berlandieri × Vitis rupestris) foi submetida à retirada de 12 ramos de ano, com no máximo 12 gemas cada, sendo subdivididos em três porções 4 gemas estaca-1) e submetidos a avaliações destrutivas e não destrutivas. As destrutivas consistiram em determinar massas fresca e seca, comprimento, diâmetro de entrenós e percentual de amido. As avaliações não estrutivas consistiram em testar o potencial de brotação e desenvolvimento das estacas. Cada planta matriz dos canteiros I e NI foi considerada uma repetição, totalizando 360 estacas por tratamento. As estacas das plantas infestadas tiveram uma redução (P<0,05), em relação às não infestadas, em diâmetro, comprimento e massas fresca e seca. Não houve contraste significativo do percentual de amido avaliado das estacas. Quanto à brotação, destaca-se que todas as estacas apresentaram o mesmo percentual (100%), independente da origem, sem alterações no desenvolvimento e crescimento das mudas. Diante desses resultados, salienta-se que videiras ?Paulsen 1103? infestadas por pérola-da-terra produzem estacas menores, porém não há comprometimento no percentual de brotação e desenvolvimento das mudas, quando comparadas com plantas não infestadas. Palavras-chave: Margarodidae, Vitaceae, interação inseto-planta, carboidrato

Robustness of germination analysis methods for Copaifera langsdorffii Desf. (Fabaceae) seeds.

2016

A concept for the viticulture of 'tropical wines'.

At worldwide level, the classical viticulture produces wines is almost all places in different climate types, where it is possible to obtain only one harvest per year. In these conditions, bud burst in vines occurs as a result of temperature raising at the end of the winter?beginning of spring. With the development of the vegetative cycle, grapes arrive to maturity/harvest period at the end of the summer?beginning of the fall season. After the fall of the leaves, vines undergo a dormant period with a vegetative repose (condition found also in some intertropical producer regions). A lot of grapes are produced for a long-time in the intertropical zone, but only a little part is used to winemaking. Considered a challenge in the past, to produce quality wines in the tropics became reality. The present industry of fine wines began about 30 years ago. Today, there are many commercial wineries in several countries in Occident and Orient in the tropical zone of the globe, like in Brazil, India, Thailand and Venezuela, producing some million liters of fine wines per year - called ?tropical wines?. This article analysis these climates, in relation to the particular viticulture adapted and developed, where it is possible to have more than one cycle per year, with one or more harvests per year. Based on the particularities of this viticulture, a characterization is proposed for the viticulture of ?tropical wines?. Some examples showing different tropical climates are presented.

Caracterização estrutural e funcional da subfamília gênica ARR-tipo B em macieira

A produtividade de pomares de macieira está estritamente relacionada ao processo de superação da dormência, o qual se caracteriza pela inabilidade do crescimento meristemático mesmo sob condições favoráveis. Embora eventos fisiológicos deste processo tenham sido elucidados, aspectos moleculares ainda são pouco compreendidos. A busca por elementos cis de regulação em genes DAM (Dormancy Associated MADS-box) de macieira revelou a presença de sítios de ligação a fatores de transcrição denominados Arabidopsis Response Regulators (ARR)?tipo B. Estes fatores fazem parte da via de sinalização de citocininas e seu papel na dormência ainda não foi elucidado. Pelo presente trabalho, temos por objetivo compreender a estrutura e a função dos elementos cis e trans associados aos fatores ARR-tipo B de macieira e avaliar se os mesmos podem estar atuando como possíveis repressores do estado dormente da planta.