Towards modification of Medicago truncatula epigenome: genome editing with engineered nucleases

Periodic drought is the primary limitation of plant growth and crop yield. The rise of water demand caused by the increase in world population and climate change, leads to one of the biggest challenges of modern agriculture: to increase food and feed production. De novo DNA methylation is a process regulated by small interfering RNA (siRNAs), which play a role in plant response and adaptation to abiotic stress. In the particular case of water deficit, growing evidences suggest a link between the siRNA pathways and drought response in the model legume Medicago truncatula. As a first step to understand the role of DNA methylation under water stress, we have set up several bioinformatics and molecular methodologies allowing the design of Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 systems and the assembly of TALENs (transcription activator-like effector nucleases), to target both dicer-like 3 (MtDCL3) and RNA-Dependent RNA polymerase (MtRDR2), enzymes of the RNA-directed DNA methylation pathway. TALENs efficiency was evaluated prior to plant transformation by a yeast-based assay using two different strategies to test TALENs activity: Polyacrylamide gel electrophoresis (PAGE) and Single strand conformation polymorphisms (SSCP). In this assay, yeast cells triple transformation emerged as good and rapid alternative to laborious yeast mating strategies. PAGE analysis might be a valuable tool to test TALENs efficacy in vivo if we could increase TALENs activity. SSCP-based approach proved to be ineffective due to the generation of several false positives. TALENs and CRISPR/Cas9 system constructed and designed in this work will in the future certainly enable the successful disruption of DCL3 and RDR2 genes and shed the light on the relationship between plant stress resistance and epigenetic regulation mediated by siRNAs in M.truncatula.

Electrical characterization and modification of low dimensional oxide semiconductors for sensor applications

This work reviews the recent research on ion and UV irradiation of β-

Bioactive beads for local sensing of proteases in 3D engineered tissues

Dissertação para obtenção do Grau de Doutor em Bioengenharia (MIT)

Differential exploitation of cashew - a low conflict crop - by sympatric humans and chimpanzees

Modification of natural areas by human activities mostly has a negative impact on wildlife by increasing the geographical and ecological overlap between people and animals. This can result in escalating levels of competition and conflict between humans and wildlife, for example over crops. However, data on specific crops and crop parts that are unattractive to wildlife yet important for human livelihoods are surprisingly scarce, especially considering their potential application to reducing crop damage by wildlife. Here we examine the co-utilization of a nationally important and spatially abundant cash crop, cashew Anacardium occidentalis, by people and chimpanzees Pan troglodytes verus inhabiting a forested–agricultural matrix in Cantanhez National Park in Guinea-Bissau. In this Park people predominantly harvest the marketable cashew nut and discard the unprofitable fruit whereas chimpanzees only consume the fruit. Local farmers generally perceive a benefit of raiding by chimpanzees as they reportedly pile the nuts, making harvesting easier. By ensuring that conflict levels over crops, especially those with high economic importance, remain low, the costs of living in proximity to wildlife can potentially be reduced. Despite high levels of deforestation associated with cashew farming, these findings point to the importance of cashew as a low-conflict crop in this area.

Evaluation of mechanical soft-abrasive blasting and chemical cleaning methods on alkyd-paint graffiti made on carbonate stones

In the context of this dissertation several studies were developed resulting in submission and publication “Evaluation of mechanical soft-abrasive blasting and chemical cleaning methods on alkyd-paint graffiti made on calcareous stones” to Journal of Cultural Heritage. (http://dx.doi.org/10.101 /j.culher.2014.10.004)

Marketing plan for online aqctivity of Rota Vicentina boosting direct website sales

The nature tourism experienced a great expansion of its market with the appearance of different lifestyles. In this Work Project a study regarding the website direct sales of Rota Vicentina was developed. Its website shows the idea of being solely an information structure and not a purchase one, leading to a current absence of online sales. Hence, it is suggested the modification of its business model, using different instruments and channels. Some digital marketing recommendations were developed in order to boost website sales, such as a platform for online reviews, remarketing campaigns and social media activity.