Relatório de Consultadoria Técnico-Científico INNEA-Alternativas Biotecnológicas Valladolid, Espanha

Entre os dias 30 de janeiro e 4 de fevereiro de 2018 foi realizada uma consultadoria técnico-científica a empresa espanhola INNEA Alternativas Biotecnológicas, com sede social na cidade de Valladolid, Espanha. Esta empresa consta de um laboratório de cultura in vitro de espécies lenhosas na localidade de Dueñas, Palencia, onde efetivamente decorreram os trabalhos. O objetivo da visita foi prestar um serviço de consultadoria na cultura in vitro de espécies lenhosas de interesse comercial para esta empresa espanhola, particularmente na micropropagação do pinheiro manso (Pinus pinea L.). O convite foi efetuado pela experiência que a equipe da Universidade de Évora formada por formada por docentes do Departamento de Biologia (Professores Amely Zavattieri, Celeste Santos Silva, Luís Silva Dias, Paulo de Oliveira e Isabel Pereira† ) e do Departamento de Química (Professores Ana Teresa Caldeira, Rosário Martins, Dora Teixeira), dos aluno de doutoramento Rogério Louro, dos doutores Carla Ragonezi e Mário Rui Castro, técnicos superiores do Departamento de Biologia (Otília Miralto e Elsa ganhão) e consultora externa Krystyna Klimaszewska do Forest Service, Canada, possuem na organogénese de pinheiro manso a partir de cotilédones de sementes maturas e a micorrização in vitro com fungos ectomicorrízicos. Estes trabalhos que decorreram por mais de uma década, contaram com a dedicação de uma numerosa equipa de investigação. Entre outros motivos, o objeto da visita foi também falar da patente a as suas possibilidades de aplicação comercial. O nosso sistema de co-cultura foi patenteado (PT 105239, INPI) pelo grande interesse para aplicação entre as co-culturas plantas-fungos e para a realização de diversos estudos bioquímicos das relações plantas simbióticas, esta patente recebeu reconhecimento internacional, em este caso particular foi um convite para realizar uma consultadoria numa empresa do setor da produção de plantas

Biotización del Pino Piñoreo (Pinus pinea L)

Apresentação didáctica sobre o tema "Biotização, feita no INTA Bella Vista, Corrientes, Argentina ao pessoal técnico-científico durante a Sabática de 2015 nessa Instituiçã

Histological Studies of Mycorrhized Roots and Mycorrhizal-Like-Structures in Pine Roots

Several studies have shown the potential of using Ectomycorrhizal (ECM) fungi in conifer micropropagation to overcome the cessation of adventitious root development. In vitro inoculation promotes the re-growth of the root system induced previously by auxin treatments, facilitating acclimation and diminishing the losses of plants because of a weak root system that is incapable of water and nutrient absorption. During a series of mycorrhization experiments, cryostat and ultrafine cuts were used to study the morpho-histological transformation of the symbiotic roots. To obtain cryostat cuts from pine roots a method frequently used for animal tissue was adopted. Molecular methods allowed fungi identification in all the mycorrhization phases and in the acclimation of derived plants. Mycorrhizal-like-structures derived from in vitro culture and axenic liquid cultures of roots were microscopically analyzed and compare with mycorrhizal roots

Relatório de Actividades - Sabática 2015

Relatório das actividades realizadas na Estação do Instituto Nacional de Tecnologia Agropecuária (INTA), Bella Vista, Corrientes, Argentina no ano de 201

Adventitious rooting of conifers: influence of biological factors

Vegetative propagation of superior conifer trees can be achieved e.g. through rooted cuttings or rooted microshoots, the latter predominantly through in vitro tissue culture. Both techniques are used to achieve rapid multiplication of trees with favorable genetic combinations and to capture a large proportion of the genetic diversity in a single generation cycle. However, adventitious rooting of shoots (cuttings) is often not efficient due to various problems such as scarcity of roots and cessation of their growth, both of which limit the application of vegetative propagation in some conifer species. Many factors are involved in the adventitious rooting of shoots including physical and chemical ones such as plant growth regulators, carbohydrates, light quality, temperature and rooting substrates or media (reviewed by Ragonezi et al. 2010). The focus of this review is on biological factors, such as inoculations with Agrobacterium rhizogenes, plant- growth-promoting rhizobacteria and other endophytes, and mycorrhizal fungi, which were found to stimulate adventitious rooting. These microorganisms could contribute not only to adventitious root development but also help in protecting conifer plants against pathogenic microorganisms, facilitate acclimation and transplanting, and contribute to more sustainable, chemical-free forests

Juniperus navicularis - a study of Portuguese population structure and fitness

Juniperus navicularis Gand. is a dioecious endemic conifer that constitutes the understory of seaside pine forests in Portugal, areas currently threatened by increasing urban expansion. The aim of this study is to assess the conservation status of previously known populations of this species located on its core area of distribution. The study was performed in south-west coast of Portugal. Three populations varying in size and pine density were analyzed. Number of individuals, population density, spatial distribution and individual characteristics of junipers were estimated. Female cone, seed characteristics and seed viability were also evaluated. Results suggest that J. navicularis populations are vulnerable because seminal recruitment is scarce, what may lead to a reduction of genetic variability due solely to vegetative propagation. This vulnerability seems to be strongly determined by climatic constraints toward increasing aridity. Ratio between male and female shrubs did not differ from 1:1 in any population. Deviations from 1:1 between mature and non-mature plants were found in all populations, denoting population ageing. Very low seed viability was observed. A major part of described Juniperus navicularis populations have disappeared through direct habitat loss to urban development, loss of fitness in drier and warmer locations and low seed viability. This study is the first to address J. navicularis conservation, and represents a valuable first step toward this species preservation

Remediation strategies in South Portugal reservoirs

A seguinte apresentação foi apresentada feita durante o Workshop On Observations in atmospheric and water sciences durante o Projecto Alex. Problemas das Albufeiras do Sul de Portugal e medidas para a sua remediação

Effects of carbon source, carbon concentration and culture conditions on in vitro rooting of Pinus pinea L. microshoots

In stone pine (Pinus pinea L.), clonal propagation via adventitious shoot formation from cotyledons has been reported before but rooting of these shoots is poor. The number of rooted shoots had low frequency limiting the used of micropropagation protocols as a choice for mass propagation of superior genotypes. Therefore the main objective of the present work was to increase the number and quality of roots per shoot in order to ensure the survival and growth of a great number of plants in the acclimation phase. To achieve this, different combinations of carbon source (sucrose or glucose) at different concentrations, under different environmental conditions (temperature and light), were tested in the induction and expression phases of the adventitious root formation. Shoots of different clones obtained via organogenesis have been employed for the experiments. Observations were made on the rooting percentage, root length and number of roots per shoot. The results showed a general increased of the number of roots per shoot and an earlier root formation when glucose was used as a carbon source. However, there were no differences in the percentage of rooting between the carbons sources tested. The best results were obtained using 0.117 M of glucose and dark treatment combined with 19°C during the induction phase of the rhizogenic process. Light and low sugar concentration proved to be beneficial for the expression phase, increasing the root length. A remarkable interclonal difference in the ability to form roots was observed. It was possible to obtain a rooting percentage of more than 75% in several of the tested clones

Adventitious rooting in olive (Olea europaea L.) cuttings: A histological evaluation

Mesmo quando estimulada pela aplicação exógena de reguladores de crescimento, a capacidade de enraizamento da oliveira, por estacaria semi-lenhosa, encontra-se muito dependente do genótipo utilizado. É assim possível encontrar cultivares como a ‘Cobrançosa’, onde se atingem sem dificuldade taxas de formação de raízes adventícias próximas dos 70%, e cultivares como a “Galega vulgar”, onde só muito esporadicamente se conseguem ultrapassar os 10% de estacas enraizadas. Este trabalho apresenta os resultados preliminares de um estudo de histologia comparativa entre as cultivares de Olea europaea referidas. Procuraram-se diferenças tanto na anatomia do caule como na ontogénese das raízes adventícias, que pudessem ajudar a compreender a diferença de comportamento das cultivares relativamente ao enraizamento por estacaria semi-lenhosa. Das observações que até agora foi possível efectuar, constatou-se que a anatomia do caule na zona de formação das novas raízes adventícias não difere significativamente, apresentando no entanto qualquer uma das cultivares em estudo um anel de esclerênquima, mais ou menos continuo, que poderá actuar como uma barreira mecânica ao normal desenvolvimento das raízes.

Sustained in vitro root development obtained in Pinus pinea inoculated with ectomycorrhizal fungi.

Stone pine (Pinus pinea L.) is an economically important forest tree in Mediterranean climates and has been the target for selection efforts through micropropagation. Previous attempts on microshoots, derived from mature seed cotyledons, reached incipient rooting after induction with a combination of auxin and hypertonic shock, but their development in vitro was not sustained. At this stage, co-culturing plantlets with some fungi isolated from ectomycorrhizas succeeded in overcoming this barrier, enabling satisfactory development in vermiculite and later in soil. About half of the fungal isolates tested helped the plants resume root growth. Although control plants (in the absence of fungi) developed roots at a later stage, i.e. during the post-transplanting acclimation in vermiculite, their growth was weaker. The root systems of some inoculated plants had ectomycorrhizas from the introduced fungi being carried over when the plants were transferred from the co-cultures to vermiculite. In conclusion, co-culturing rooted microshoots with ectomycorrhizal fungi can be an effective means to overcome the difficulties encountered in the use of micropropagation methods on this species