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.

Caracterização do cDNA da NAD-MDH citosólica e da H+ ATPase Vacuolar no amadurecimento da anona (Annona cherimola Mill)

A Annona cherimola é um fruto exótico, com um sabor agradável. Este fruto tem um elevado potencial comercial, mas apresenta um tempo médio de vida curto devido ao seu rápido amadurecimento. Por esta razão é necessário conhecer melhor o processo de amadurecimento deste fruto. Na Região Autónoma da Madeira a cultura da anoneira é muito importante em termos comerciais. O processo de amadurecimento leva a diversas modificações bioquímicas e fisiológicas. Existem várias enzimas e substâncias que integram este processo. Neste trabalho iremos estudar os genes das enzimas malato desidrogenase e H+ ATPase vacuolar que estão envolvidos no processo de amadurecimento dos frutos. Utilizando as técnicas de RACE e sequenciação foi possível determinar a sequência nucleotídica do cDNA destes genes. O cDNA da malato desidrogenase é composto por 1364 nucleótidos, contendo uma zona 5’ UTR com 84 nucleótidos, uma zona 3’ UTR com 284 nucleótidos e um sinal de poliadenilação com a sequência AATAAA. A ORF apresenta 996 nucleótidos, codificando uma proteína com 332 aminoácidos. Para a H+ ATPase vacuolar foi amplificado o cDNA da subunidade C do domínio V1. Esta apresenta 799 nucleótidos, dos quais 36 são da 5’ UTR, 266 da 3’ UTR e 498 da ORF. A ORF codifica uma proteína com 166 aminoácidos.

Comparative analysis of the volatile fraction from Annona cherimola Mill. cultivars by solid-phase microextraction and gas chromatography–quadrupole mass spectrometry detection

The analysis of volatile compounds in Funchal, Madeira, Mateus and Perry Vidal cultivars of Annona cherimola Mill. (cherimoya) was carried out by headspace solid-phase microextraction (HS-SPME) combined with gas chromatography–quadrupole mass spectrometry detection (GC–qMSD). HS-SPME technique was optimized in terms of fibre selection, extraction time, extraction temperature and sample amount to reach the best extraction efficiency. The best result was obtained with 2 g of sample, using a divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS) fibre for 30 min at 30 °C under constant magnetic stirring (800 rpm). After optimization of the extraction methodology, all the cherimoya samples were analysed with the best conditions that allowed to identify about 60 volatile compounds. The major compounds identified in the four cherimoya cultivars were methyl butanoate, butyl butanoate, 3-methylbutyl butanoate, 3-methylbutyl 3-methylbutanoate and 5-hydroxymethyl-2-furfural. These compounds represent 69.08 ± 5.22%, 56.56 ± 15.36%, 56.69 ± 9.28% and 71.82 ± 1.29% of the total volatiles for Funchal, Madeira, Mateus and Perry Vidal cultivars, respectively. This study showed that each cherimoya cultivars have 40 common compounds, corresponding to different chemical families, namely terpenes, esters, alcohols, fatty acids and carbonyl compounds and using PCA, the volatile composition in terms of average peak areas, provided a suitable tool to differentiate among the cherimoya cultivars.

Integrated valorization of Anona Cherimola Mill. seeds

Agricultural and agro-industrial residues are often considered both an environmental and an economical problem. Therefore, a paradigm shift is needed, assuming residues as biorefinery feedstocks. In this work cherimoya (Annona cherimola Mill.) seeds, which are lipid-rich (ca. 30%) and have a significant lignocellulosic fraction, were used as an example of a residue without any current valorization. Firstly, the lipid fraction was obtained by solvent extraction. Extraction yield varied from 13% to 28%, according to the extraction method and time, and solvent purity. This oil was converted into biodiesel (by base-catalyzed transesterification), yielding 76 g FAME/100 g oil. The obtained biodiesel is likely to be incorporated in the commercial chain, according to the EN14214 standard. The remaining lignocellulosic fraction was subjected to two alternative fractionation processes for the selective recovery of hemicellulose, aiming different products. Empirical mathematical models were developed for both processes, aiming future scale-up. Autohydrolysis rendered essentially oligosaccharides (10 gL-1) with properties indicating potential food/feed/pharmacological applications. The remaining solid was enzymatically saccharified, reaching a saccharification yield of 83%. The hydrolyzate obtained by dilute acid hydrolysis contained mostly monosaccharides, mainly xylose (26 gL-1), glucose (10 gL-1) and arabinose (3 gL-1), and had low content of microbial growth inhibitors. This hydrolyzate has proven to be appropriate to be used as culture media for exopolisaccharide production, using bacteria or microbial consortia. The maximum conversion of monosaccharides into xanthan gum was 0.87 g/g and kefiran maximum productivity was 0.07 g.(Lh)-1. This work shows the technical feasibility of using cherimoya seeds, and materials as such, as potential feedstocks, opening new perspectives for upgrading them in the biorefinery framework.

Novo sistema hidropónico aplicado à produção do tomate cereja (Lycopersicum esculentum Mill. var Moscatel RZ)

A hidroponia é uma técnica poderosa na utilização eficiente dos recursos agrícolas, pois permite um ganho quer na produtividade, quer na qualidade do fruto. Atualmente os sistemas hidropónicos utilizados em plantas de médio porte não são os mais adequados, sendo esta uma área a explorar. Neste trabalho criou-se um novo sistema hidropónico (Deep Large Flow Technique- DLFT) para aplicação a plantas de médio porte e avaliou-se o seu impacto no crescimento, produtividade, biomassa e qualidade do tomate cereja (Solanum lycopersicum var. Moscatel RZ). O sistema de cultivo tradicional (solo) e semi-hidropónico (suporte com fibra de coco) serviram como base de comparação. Observou-se que após 31 dias as plantas cultivadas no novo sistema hidropónico obtiveram um crescimento acentuado, apresentando 3x mais frutos. Em termos de produtividade, os sistemas hidropónicos foram iguais, sendo o sistema tradicional 4x inferior. O total de biomassa foi significativamente maior no novo sistema hidropónico, com mais 20% e 88% que o sistema semi-hidropónico e tradicional, respetivamente. As plantas produzidas no sistema DLFT apresentaram frutos com qualidade superior, com um rácio de monossacarídeos/acidez de 6,6 mg/g. O conteúdo total de ácidos gordos nos frutos cultivados com este sistema foi 39% e 44% superior aos do semihidropónico e do tradicional. Contrariamente, o conteúdo de flavonóides foi inferior nos frutos cultivados com o novo sistema hidropónico, tendo os frutos do sistema tradicional e do sistema semi-hidropónico 40% e 10% maior teor desta família de compostos. Os resultados obtidos neste estudo demonstram que a utilização do novo sistema hidropónico na produção de plantas de médio porte, aumenta o crescimento, a acumulação de biomassa e a qualidade do fruto. Assim, permite ao produtor reduzir os custos, rentabilizar a produção (menor tempo de produção), valorizar a biomassa da planta e aumentar a qualidade do produto. Também, através dos teores de carotenóides, ácidos gordos e polifenóis poderá inferir-se que ocorreu um menor impato dos stressses abióticos subjacentes aos sistemas nas plantas produzidas pelo sistema DLFT.