Genetic characterization of Portuguese allochthonous populations of Pinus nigra using ISSRs and SCoTs and extrapolation of their infraspecific taxonomy.

The Pinus nigra distribution in Portugal is restricted to six allochthonous populations with unknown origin and infraspecific taxonomy. This work intends to evaluate their genetic diversity, structure and relationships, and to infer about their infraspecific taxonomy by comparing molecular patterns produced by inter-simple sequence repeat and Start Codon Targeted markers among Portuguese and foreign samples with known taxonomy and provenance. 127 Portuguese P. nigra individuals were clustered per population. The genetic differentiation was higher within rather than among populations. The pooled molecular data indicated high genetic proximity among the Portuguese and foreign samples of subspecies laricio. However, the separate analysis per marker system demonstrated that two varieties of subspecies laricio (corsicana and calabrica) may have been used in the plantations of the Portuguese P. nigra stands performed in the last century. The genetic characterization and extrapolation of the intraspecific taxonomy of these populations provide useful information for forest management, afforestation and germplasm use.

A comprehensive assessment of the transcriptome of cork oak (Quercus suber) through EST sequencing.

BACKGROUND: Cork oak (Quercus suber) is one of the rare trees with the ability to produce cork, a material widely used to make wine bottle stoppers, flooring and insulation materials, among many other uses. The molecular mechanisms of cork formation are still poorly understood, in great part due to the difficulty in studying a species with a long life-cycle and for which there is scarce molecular/genomic information. Cork oak forests are of great ecological importance and represent a major economic and social resource in Southern Europe and Northern Africa. However, global warming is threatening the cork oak forests by imposing thermal, hydric and many types of novel biotic stresses. Despite the economic and social value of the Q. suber species, few genomic resources have been developed, useful for biotechnological applications and improved forest management. RESULTS: We generated in excess of 7 million sequence reads, by pyrosequencing 21 normalized cDNA libraries derived from multiple Q. suber tissues and organs, developmental stages and physiological conditions. We deployed a stringent sequence processing and assembly pipeline that resulted in the identification of ~159,000 unigenes. These were annotated according to their similarity to known plant genes, to known Interpro domains, GO classes and E.C. numbers. The phylogenetic extent of this ESTs set was investigated, and we found that cork oak revealed a significant new gene space that is not covered by other model species or EST sequencing projects. The raw data, as well as the full annotated assembly, are now available to the community in a dedicated web portal at http://www.corkoakdb.org. CONCLUSIONS: This genomic resource represents the first trancriptome study in a cork producing species. It can be explored to develop new tools and approaches to understand stress responses and developmental processes in forest trees, as well as the molecular cascades underlying cork differentiation and disease response.

Implementation of the e-Bug Project in Portugal.

In Portugal, the need for health educational campaigns to reduce antibiotic misuse, and the implementation of handwashing and hygiene measures to reduce cross-transmission have become national priorities. The implementation of the e-Bug Project in our country has been conducted by the Directorate-General for Health as the beneficiary entity of the Project, in partnership with the Ministry of Education. The involvement of the operational structures of both ministries in the development of the Project has been guaranteed from the beginning. We have translated and produced 6000 junior and senior e-Bug packs, and started to distribute them to science teachers, health education coordinators and school health teams from all over the country, who were identified as stakeholders and registered as e-Bug users in an e-learning platform from the Ministry of Education. Portugal is deeply interested in the long-term sustainability of the e-Bug Project. Portuguese schools and the Portuguese e-Bug team will continue to support this initiative to benefit the health education of Portuguese children.

A national e-Delphi towards the measurement of safe medication practices in Portuguese hospitals.

OBJECTIVES: To determine the face and content validity of items for measuring safe medication practices in Portuguese hospitals. METHODS: 128 items were drafted from content analysis of existing questionnaires and the literature, employing preferred terms of the WHO International Classification for Patient Safety (Portuguese version). A two-round e-Delphi was convened, using a purposive multidisciplinary panel. Hospital-based experts were asked to rate the relevance of items on a 7-point Likert scale and to comment on their clarity and completeness. RESULTS: The response rate was similar in both rounds (70.3% and 73.4%, respectively). In the first round 91/128 (71.1%) items reached the predefined level of positive consensus. In the second round 23 additional items reached positive consensus, as well as seven items newly derived by the panel. CONCLUSIONS: Most items have face and content validity, indicating relevance and clarity, and can be included in a future questionnaire for measuring safe medication practices in Portuguese hospitals.

Differential physiological and genetic responses of five European Scots pine provenances to induced water stress.

Pinus sylvestris L. (Scots pine) is the conifer with widest natural distribution area. Portugal constitutes its westernmost limit of distribution. Most of the Portuguese populations were planted but two autochthonous populations were recently ascribed to 'Serra do Gerês' (NW Portugal), and seem to be well adapted to the temperate climate. However, the ongoing climate changes may compromise their survival. With this study we intend to evaluate the anatomic-physiological and genetic responses of Scots pine from five European provenances ('Gerês', 'Puebla de Lillo', 'Montes Universales', Germany and Sweden) to three water availability regimes, in order to determine which one(s) present higher resistance to drought. Individuals from 'Gerês' presented the highest stability in photosynthetic reactions as well as the better photochemical and metabolic behaviours under drought (T3). Hence, the relative expression ratio of three water stress-responsive genes during drought was lower and gradual in 'Gerês', compared to all other provenances, followed by Germany. The results achieved in 'Gerês' and Germany provenances are very interesting since they reflected that the native populations of 'Gerês' along with the Portuguese Scots pine planted populations with a probable German provenance, have ability and high adaptive potential to respond to situations of water deficit. Moreover, the present genetic and physiological data demonstrated the urgent demand for the conservation of Portuguese Scots pine genetic resources as well as its use in plantation/afforestation of areas where the warming and drought has been affecting the survival of this species.

Genetic variation of drought tolerance in Pinus pinaster at three hierarchical levels: a comparison of induced osmotic stress and field testing.

Understanding the survival capacity of forest trees to periods of severe water stress could improve knowledge of the adaptive potential of different species under future climatic scenarios. In long lived organisms, like forest trees, the combination of induced osmotic stress treatments and field testing can elucidate the role of drought tolerance during the early stages of establishment, the most critical in the life of the species. We performed a Polyethylene glycol-osmotic induced stress experiment and evaluated two common garden experiments (xeric and mesic sites) to test for survival and growth of a wide range clonal collection of Maritime pine. This study demonstrates the importance of additive vs non additive effects for drought tolerance traits in Pinus pinaster, and shows differences in parameters determining the adaptive trajectories of populations and family and clones within populations. The results show that osmotic adjustment plays an important role in population variation, while biomass allocation and hydric content greatly influence survival at population level. Survival in the induced osmotic stress experiment presented significant correlations with survival in the xeric site, and height growth at the mesic site, at population level, indicating constraints of adaptation for those traits, while at the within population level no significant correlation existed. These results demonstrate that population differentiation and within population genetic variation for drought tolerance follow different patterns.