The Rg1 allele as a valuable tool for genetic transformation of the tomato 'Micro-Tom' model system

Background: The cultivar Micro-Tom (MT) is regarded as a model system for tomato genetics due to its short life cycle and miniature size. However, efforts to improve tomato genetic transformation have led to protocols dependent on the costly hormone zeatin, combined with an excessive number of steps. Results: Here we report the development of a MT near-isogenic genotype harboring the allele Rg1 (MT-Rg1), which greatly improves tomato in vitro regeneration. Regeneration was further improved in MT by including a two-day incubation of cotyledonary explants onto medium containing 0.4 mu M 1-naphthaleneacetic acid (NAA) before cytokinin treatment. Both strategies allowed the use of 5 mu M 6-benzylaminopurine (BAP), a cytokinin 100 times less expensive than zeatin. The use of MT-Rg1 and NAA pre-incubation, followed by BAP regeneration, resulted in high transformation frequencies (near 40%), in a shorter protocol with fewer steps, spanning approximately 40 days from Agrobacterium infection to transgenic plant acclimatization. Conclusions: The genetic resource and the protocol presented here represent invaluable tools for routine gene expression manipulation and high throughput functional genomics by insertional mutagenesis in tomato.

Convergence of developmental mutants into a single tomato model system: 'Micro-Tom' as an effective toolkit for plant development research

Background: The tomato (Solanum lycopersicum L.) plant is both an economically important food crop and an ideal dicot model to investigate various physiological phenomena not possible in Arabidopsis thaliana. Due to the great diversity of tomato cultivars used by the research community, it is often difficult to reliably compare phenotypes. The lack of tomato developmental mutants in a single genetic background prevents the stacking of mutations to facilitate analysis of double and multiple mutants, often required for elucidating developmental pathways. Results: We took advantage of the small size and rapid life cycle of the tomato cultivar Micro-Tom (MT) to create near-isogenic lines (NILs) by introgressing a suite of hormonal and photomorphogenetic mutations (altered sensitivity or endogenous levels of auxin, ethylene, abscisic acid, gibberellin, brassinosteroid, and light response) into this genetic background. To demonstrate the usefulness of this collection, we compared developmental traits between the produced NILs. All expected mutant phenotypes were expressed in the NILs. We also created NILs harboring the wild type alleles for dwarf, self-pruning and uniform fruit, which are mutations characteristic of MT. This amplified both the applications of the mutant collection presented here and of MT as a genetic model system. Conclusions: The community resource presented here is a useful toolkit for plant research, particularly for future studies in plant development, which will require the simultaneous observation of the effect of various hormones, signaling pathways and crosstalk.

Hormonal modulation of photomorphogenesis-controlled anthocyanin accumulation in tomato (Solanum lycopersicum L. cv Micro-Tom) hypocotyls: Physiological and genetic studies

Hormones are likely to be important factors modulating the light-dependent anthocyanin accumulation. Here we analyzed anthocyanin contents in hypocotyls of near isogenic Micro-Tom (MT) tomato lines carrying hormone and phytochrome mutations, as single and double-mutant combinations. In order to recapitulate mutant phenotype, exogenous hormone applications were also performed Anthocyanin accumulation was promoted by exogenous abscisic acid (ABA) and inhibited by gibberellin (GA), in accordance to the reduced anthocyanin contents measured in ABA-deficient (notabills) and GA-constitutive response (procera) mutants. Exogenous cytokinin also enhanced anthocyanin levels in MT hypocotyls. Although auxin-insensitive chageotropica mutant exhibited higher anthocyanin contents, pharmacological approaches employing exogenous auxin and a transport inhibitor did not support a direct role of the hormone in anthocyanin accumulation Analysis of mutants exhibiting increased ethylene production (epwastic) or reduced sensitivity (Never ripe), together with pharmacological data obtained from plants treated with the hormone, indicated a limited role for ethylene in anthocyanin contents. Phytochrome-deficiency (aurea) and hormone double-mutant combinations exhibited phenotypes suggesting additive or synergistic interactions, but not fully espistatic ones, in the control of anthocyanin levels in tomato hypocotyls. Our results indicate that phytochrome-mediated anthocyanin accumulation in tomato hypocotyls is modulated by distinct hormone classes via both shared and independent pathways. (C) 2010 Elsevier Ireland Ltd. All rights reserved

The isolation of antioxidant enzymes from mature tomato (cv. Micro-Tom) plants

The activity of catalase (CAT), guaiacol peroxidase (GPOX), ascorbate peroxidase (APX), glutathione reductase (GR), and the isoenzymes of superoxide dismutase (SOD) were determined in the organs of tomato (Lycopersicon esculentum) cultivar Micro-Tom after 104 days of development. The total activities of CAT, GPOX, and GR were higher in the stem than in others tissues, whereas the stem exhibited the lowest APX activity. Activity staining analysis following gel electrophoresis revealed the existence of four SOD isoenzymes in leaves, three in fruits, but only two in the roots and stems. This characterization is essential for an investigation into the effect of abiotic and biotic stresses on the oxidative stress responses by this plant model system.

Acquired tolerance of tomato (Lycopersicon esculentum cv. Micro-Tom) plants to cadmium-induced stress

The effects of varying concentrations of cadmium (Cd) on the development of Lycopersicon esculentum cv. Micro-Tom (MT) plants were investigated after 40 days (vegetative growth) and 95 days (fruit production), corresponding to 20 days and 75 days of exposure to CdCl(2), respectively. Inhibition of growth was clearly observed in the leaves after 20 days and was greater after 75 days of growth in 1 mM CdCl(2), whereas the fruits exhibited reduced growth following the exposure to a concentration as low as 0.1 mM CdCl(2). Cd was shown to accumulate in the roots after 75 days of growth but was mainly translocated to the upper parts of the plants accumulating to high concentrations in the fruits. Lipid peroxidation was more pronounced in the roots even at 0.05 mM CdCl(2) after 75 days, whereas in leaves, there was a major increase after 20 days of exposure to 1 mM CdCl(2), but the fruit only exhibited a slight significant increase in lipid peroxidation in plants subjected to 1 mM CdCl(2) when compared with the control. Oxidative stress was also investigated by the analysis of four key antioxidant enzymes, which exhibited changes in response to the increasing concentrations of Cd tested. Catalase (EC 1.11.1.6) activity was shown to increase after 75 days of Cd treatment, but the major increases were observed at 0.1 and 0.2 mM CdCl(2), whereas guaiacol peroxidase (EC 1.11.1.7) did not vary significantly from the control in leaves and roots apart from specific changes at 0.5 and 1 mM CdCl(2). The other two enzymes tested, glutathione reductase (EC 1.6.4.2) and superoxide dismutase (SOD, EC 1.15.1.1), did not exhibit any significant changes in activity, apart from a slight decrease in SOD activity at concentrations above 0.2 mM CdCl(2). However, the most striking results were obtained when an extra treatment was used in which a set of plants was subjected to a stepwise increase in CdCl(2) from 0.05 to 1 mM, leading to tolerance of the Cd applied even at the final highest concentration of 1 mM. This apparent adaptation to the toxic effect of Cd was confirmed by biomass values being similar to the control, indicating a tolerance to Cd acquired by the MT plants.

A água, o saneamento básico e os residuos sólidos urbanos no desenvolvimento de São Tomé e Príncipe

RESUMO: O trabalho que se apresenta no âmbito desta dissertação, é direcionado para a problemática da Água, do Saneamento Básico e dos Resíduos Sólidos Urbanos «RSU» em São Tomé e Príncipe. Num contexto de desenvolvimento e indo ao encontro dos anseios da Organização das Nações Unidas «ONU» e da sua perspetiva de alcançar os Objetivos do Milénio nesta área tão importante. Elegeu-se como primordial objetivo, conhecer as indicações técnico políticas instituídas em São Tomé e Príncipe, para a gestão dos problemas acima enumerados. Entender esses problemas, identificar as dificuldades sentidas pelo governo e pela generalidade dos seus habitantes no acesso à água, ao saneamento básico, à recolha e tratamento de RSU. Outra vertente será direcionada para apontar caminhos nestas áreas, onde a capacidade institucional tarda em dar resposta às necessidades básicas destes setores, inviabilizando um desenvolvimento sustentado destes ramos. Esta dissertação, assenta ainda no reconhecimento e na importância estratégica em se valorizar e consolidar redes técnico-científicas no âmbito da Linha de Investigação em Estudos Africanos e Pós-Coloniais, inserida na Unidade de Estudos e Investigação em Ciência, Tecnologia e Sociedade «UEICTS» da Universidade Lusófona de Humanidades e Tecnologias «ULHT».ABSTRACT: The work presented in this dissertation, is directed to the problem of Water Sanitation and Solid Waste «RSU» in Sao Tome and Principe. In a context of development, fulfillment of the wishes of the United Nations «UN» and its prospect of achieving the Millennium Goals in this important area. The prime objective, is, to know the indications and technical policies in place in Sao Tome and Principe for the management of the problems listed above. Understanding these problems, identifying the difficulties faced by government and by most of its residents in relation, to access to water, sanitation, collection and treatment of Solid Waste. Another aspect, is directed, to point, to ways, in these subjects where institutional capacity is slow to respond to basic needs of these sectors, preventing a sustained development of these industries. This dissertation focus on the recognition and strategic importance in considering and consolidate technical and scientific networks in the line of Research in African and Lusophone, inserted at the Unit for Studies and Research in Science, Technology and Society «UEICTS» Lusophone University of Humanities and Technology «ULHT».

A inspecção da educação na República Democrática de São Tomé e Príncipe: concepções, dinâmicas e estruturas organizacionais

Dissertação apresentada à Escola Superior de Educação de Lisboa para Obtenção de Grau de Mestre em Ciências da Educação - Especialização em Supervisão da Educação

Que formação de professores para a inclusão em São Tomé e Príncipe?

Dissertação apresentada à escola superior de educação de lisboa para obtenção do grau de mestre em ciências da educação - especialidade educação especial

Trabalho colaborativo entre professores para integração curricular no 1º ciclo do ensino básico: um contributo para melhoria das planificações pedagógicas em S.Tomé e Príncipe

Dissertação apresentada à Escola Superior de Educação de Lisboa para obtenção do grau de mestre em Didáticas Integradas em Língua Portuguesa, Matemática, Ciências Naturais e Sociais