Unravelling the nanostructure of strawberry fruit pectins by atomic force microscopy

Atomic force microscopy (AFM) allows the analysis of individual polymers at nanostructural level with a minimal sample preparation. This technique has been used to analyse the pectin disassembly process during the ripening and postharvest storage of several fleshy fruits. In general, pectins analysed by AFM are usually visualized as isolated chains, unbranched or with a low number of branchs and, occasionally, as large aggregates. However, the exact nature of these structures is unknown. It has been suggested that pectin aggregates represent a mixture of rhamnonogalacturonan I and homogalacturonan, while isolated chains and their branches are mainly composed by polygalacturonic acid. In order to gain insight into the nature of these structures, sodium carbonate soluble pectins from ripe strawberry (Fragaria x ananassa, Duch.) fruits were subjected to enzymatic digestion with endo-Polygalacturonase M2 from Aspergillus aculeatus, and the samples visualized by AFM at different time intervals. Pectins isolated from control, non-transformed plants, and two transgenic genotypes with low level of expression of ripening-induced pectinase genes encoding a polygalacturonase (APG) or a pectate lyase (APEL) were also included in this study. Before digestion, isolated pectin chains from control were shorter than those from transgenic fruits, showing number-average (LN) contour length values of 73.2 nm vs. 95.9 nm and 91.4 nm in APG and APEL, respectively. The percentage of branched polymers was significantly higher in APG polyuronides than in the remaining genotypes, 33% in APG vs. 6% in control and APEL. As a result of the endo-PG treatment, a gradual decrease in the main backbone length of isolated chains was observed in the three samples. The minimum LN value was reached after 8 h of digestion, being similar in the three genotypes, 22 nm. By contrast, the branches were not visible after 1.5-2 h of digestion. LN values were plotted against digestion time and the data fitted to a first-order exponential decay curve, obtaining R2 values higher than 0.9. The half digestion time calculated with these equations were similar for control and APG pectins, 1.7 h, but significantly higher in APEL, 2.5 h, indicating that these polymer chains were more resistant to endo-PG digestion. Regarding the pectin aggregates, their volumes were estimated and used to calculate LN molecular weights. Before digestion, control and APEL samples showed complexes of similar molecular weights, 1722 kDa, and slightly higher than those observed in APG samples. After endo-PG digestion, size of complexes diminished significantly, reaching similar values in the three pectin samples, around 650 kDa. These results suggest that isolated polymer chains visualized by AFM are formed by a HG domain linked to a shorter polymer resistant to endo-PG digestion, maybe xylogalacturonan or RG-I. The silencing of the pectate lyase gene slightly modified the structure and/or chemical composition of polymer chains making these polyuronides more resistant to enzymatic degradation. Similarly, polygalacturonic acid is one of the main component of the aggregates.

Effect of clearing native forest (Caatinga) strips on some hydrologic components of micro-watersheds.

The influences of clearing native vegetation (Caatinga) in contour strips at 25 cm vertical interval on evaporation losses in cleared strips, annual runoff efficiency and annuall soil loss on gently sloped micro-waterheds in the arid zones of Northeast Brazil are reported. The alternate native vegetation (Caatinga) strips function very effectively as windbreaks thus reducing evaporation losses substantially in the leeward cleared strips. The runoff measured at the micro-watershed with cleared strips was many-fold lower than the runoff obtained at a completely denuded watershed even when it was protected by narrow based channel terraces. However, the annual runoff efficiency can be significantly increased in a strip cleared watershed if narrow based channel terraces are provided on the lower side of cleared strips. The annual soil losses in strip cleared watersheds as well as completely denuded waterhed of gentle slopes were negligible. Thus clearing land in alternate contour strips on a micro-watersheds shall substantially improve crop water use efficiency without creating any significant erosion problems. Additionally this treatment will increase runoff for water harvesting for irrigation purposes.

Relatório final de estágio realizado na Escola Básica Integrada Rainha Dona Leonor de Lencastre no ano letivo 2011-2012

Como parte integrante do 2º ano de Mestrado em Ensino da Educação Física nos Ensinos Básico e Secundário da Faculdade de Motricidade Humana, o presente relatório tem como finalidade analisar o processo de estágio pedagógico desenvolvido no ano letivo 2012/2013 na Escola Básica Integrada Rainha Dona Leonor de Lencastre, localizada em S. Marcos, concelho de Sintra. Este estágio tem em vista a preparação para a função docente. Todo o processo teve como referência o Guia de Estágio Pedagógico 2012/2013 onde se encontram especificados os objetivos a atingir nas quatro áreas inerentes ao processo de estágio: Organização e Gestão do Ensino e da Aprendizagem (Área 1), Inovação e Investigação Pedagógica (Área 2), Participação na Escola (Área 3) e Relação com a Comunidade (Área 4). Em relação a cada uma destas áreas será realizada uma aprofundada reflexão sobre as principais dificuldades sentidas, as estratégias utilizadas para a sua superação e os contributos finais de todo este ciclo de aprendizagem para mim enquanto professora. Em último lugar será apresentada uma reflexão global acerca de todo o processo formativo salientando as principais etapas desta longa aprendizagem e quais aquelas que considerei mais enriquecedoras para o meu futuro enquanto docente.