Efeito de extractos de raízes eliciadas de Castanea sativa no crescimento do fungo ectomicorrízico Pisolithus tinctorius

O castanheiro (Castanea sativa Mill.) estabelece associação com numerosas espécies de fungos ectomicorrízicos, estando descritos os efeitos benéficos para a planta após micorrização com Pisolithus tinctorius. Neste processo é essencial que ocorra a troca de sinais entre os simbiontes, para que seja reconhecida a sua compatibilidade e para que ocorra a formação dos órgãos ectomicorrízicos. Neste trabalho são fornecidas evidências que sugerem a capacidade de extractos de raízes de castanheiro, nos estádios iniciais de contacto com P. tinctorius regularem o crescimento do fungo micorrízico

Endo- and epiphytic fungal communities of olive twigs is influenced by cultivar and olive knot infection

Olive tree phyllosphere is colonized by a diverse microbial assemblage that may interact with pathogenic fungi, making them potential candidates for disease supression. Olive knot (OK) is a disease caused by the bacterium Pseudomonas savastanoi pv. savastanoi (Psv.) with significant economic losses in olive tree (Olea europaea L.). With this work we want to evaluate the effect of cultivar and OK infection on both endo- and epiphytic fungal community inhabiting olive twig tissues. For this, fungal composition and diversity was assessed in both asymptomatic and OK-symptomatic twigs of tree co-occurring olive cultivars with different susceptibilities to OK disease. The identification of isolated species was performed by ITS rDNA sequencing. The cultivar and OK infection were important in shaping both endophytic and epiphytic fungal communities. Fungal community composition was found to differ significantly (p=0.005) between olive tree cultivars, being Nectriaceae the dominant family in cvs. Cobrançosa and Verdeal Transmontana, whereas Pleosporaceae was dominant in the cv. Madural. Epiphytic and endophytic fungal communities also differed in size and in composition in asymptomatic and OK-symptomatic twigs, for the three cultivars. In general, asymptomatic twigs were more diverse and rich (up to 1.4-fold) when compared to OK-symptomatic twigs. Among the species identified in the asymptomatic tissues, Cladosporium cladosporioides and Chromelosporium carneum were the most frequently isolated within epiphytic and endophytic community, respectively; whereas in the OK-symptomatic tissues Cladosporium sp. and Fusarium lateritium were the most frequently isolated within epiphytic and endophytic community, respectively. According to indicator species analysis C. carneum, Pyronema dosmesticum and Phoma aloes (IndVal up to 0.56) may be promising species in the OK suppression. Better acknowledgement should be developed in order to uncover their role on plant health.FEDER funds through COMPETE (Programa Operacional Factores de Competitividade} and by national funds by FCT [Fundação para a Ciência e a Tecnologia) in the framework of the project EXCL/AGR-PRO/0591/2012. T. Gomes thanks FCT, POPH-QREN and FSE for PhD SFRH/BD/98127/2013 Grantinfo:eu-repo/semantics/publishedVersio

Barcoding of entomopathogenic fungi from olive tree pests : prospects and limitations

From all the barcoding initiatives in progress, fungal barcode is probably the one where more difficulties have been encountered. While for plants and animais the barcode regions were easy to define, for fungi the choice was not so straightforward. The internal transcribed spacer (ITS) region was one of the proposed DNA regions for barcoding fungi. This is an extensively used region, for molecular systematic and identification of species, being probably the most widely sequenced DNA region of fungi. This is due to the simplicity of the amplification, related to the multicopy nature of the rDNA; the possibility of using universal primers; and the high levei of sequence variation that occurs even between closely related species. Furthermore, a significant number of identified sequences for comparison are available in the GenBank database. Although the ITS region of rDNA was chosen for some groups of fungi, the use of this region presents very limited application for others, especially for Ascomycetes. As some of the most important entomopathogenic fungi are Ascomycetes, belonging to genera Beauveria, Cordyceps, Isaria, Lecanicillium and Paecilomyces, the use of the ITS region for barcoding purpose are being complemented with other regions. This work, based on the identification of fungal entomopathogens isolated directly from cadavers of one of the major pests in olive graves, the olive moth iPrevs oleae Bern.), intends to illustrate the application of the ITS region to identify these fungal species. The use of this region proved to be useful for the identification of most of the entomopathogenic fungi found in dead larvae and pupae of P. oleae. However, the use ot ITS region for barcode purposes did not allow the identification of several isolates, proving the requiremerít of using a second barcoding region, to enable full fungal identification.This work has been supported by FCT (PTDC/AGR-AAM/02600/2008)

Identification of fungi isolated from the olive moth (Prays oleae Bern.) based on ITS region

Fundação para a Ciência e Tecnologia (FCT) - Project PTDC/AGR-AAM/102600/200

Fungal diversity among different annual generations of Prays oleae

Comunicação em painelThe olives and olive oil production are important agricultural activities in Portugal. In the region of Trás-os-Montes, olive orchards are strongly affected by Prays oleae Bern., which is responsible for high losses in the olive production, as much as 40% of the expected yield. This lepidopteran presents three generations per year (phyllophagous, antophagous and carpophagous) that damage the olive tree in different organs (leaves, flowers and fruits, respectively). In an attempt to identify fungi that might cause the death of olive moths, larvae and pupae of those three annual generations were collected and surveyed for natural fungal infection. After their isolation, the fungal agents were molecularly identified by sequencing the amplified internal transcribed spacer (ITS) region of rDNA. In the present work, the diversity of fungal species associated to P. oleae, in several olive orchards located in Trás-os-Montes region, is discussed. The diversity and abundance of fungal species differed between all three generations. Higher diversity was found in the carpophagous generation, followed by the antophagous and phyllophagous generations. The use of already adapted fungal species to control one of the major pests of olive groves will increase the success of a future biocontrol strategy. In this context, the identification of fungi associated to P. oleae from olive orchards provided a pool of potential biocontrol agents. In this work, Beauveria bassiana proved to be the most promising fungus to be used as biocontrol agent against olive moth, being strongly associated to the phyllophagous generation. Other fungal species presenting entomopathogenic, antagonistic and phytopathogenic characteristics were also found.This work has been supported by FCT (PTDC/AGR-AAM/102600/2008)

Screening for angiotensin I-converting enzyme (ACE) inhibitory activity of enzymatic hydrolysates obtained from Azorean macroalgae

As part of ongoing studies into the isolation of antihypertensive peptides, 12 common macroalgae species from Azores were investigated for their potential angiotensinconverting enzyme (ACE) inhibitory properties. After optimization of enzymatic hydrolysis conditions, the anti-ACE activity was determined by high performance liquid chromatography (HPLC). The species under study include four Rhodophyta (Gelidium microdon, Osmundea pinnatifida, Porphyra sp. and Pterocladiella capillacea), four Chlorophyta (Chaetomorpha linum, Codium adhaerens, Ulva compressa and Ulva rigida) and four Heterokontophyta (Cystoseira humilis, Fucus spiralis, Padina pavonica and Sargassum cymosum). The bromelain hydrolysates derived from P. capillacea, Porphyra sp., F. spiralis, U. compressa, O. pinnatifida and U. rigida were the most active presenting an ACE inhibitory activity of 48.73 ± 1.58%, 50.96 ± 1.82%, 56.77 ± 1.80%, 59.80 ± 0.95%, 62.59 ± 1.60% and 65.68 ± 0.72%, respectively. These results suggest that some macroalgae from the littoral Azorean sea are potential sources of powerful ACE inhibitor peptides, with potential impact on public health, particularly on the hypertensive patients. These features make these algae valuable to the regional economy and may also provide an incentive for marine biodiversity conservation and valuation of marine algal resources

Nutritional value of selected macroalgae

Copyright © Springer Science+Business Media B.V. 2011.Macroalgae are traditionally used in human and animal nutrition. Their protein and fiber content have been widely studied and differ according to the species, their geographic origin and their seasonal conditions. In addition to their value for human nutrition, seaweeds have multiple therapeutically applications (e.g., weight control, hypocholesterolemic, antioxidant and antitumor activities, others) and, in general, contribute and promote human health. In the archipelago of the Azores, the consumption of seaweeds is widespread and accepted as a common practice in some islands. This work is aimed at providing information on the protein and fiber content of the locally consumed species, to promote this regional food product that can be potentially profitable from the biotechnology and commercial perspective, and also benefit public health, particularly, taking into account the low level of marine pollution in the Azores archipelago. Protein and fiber content of eight seaweeds (Porphyra sp., Osmundea pinnatifida, Pterocladiella capillacea, Sphaerococcus coronopifolius, and Gelidium microdon, Rhodophyta; Cystoseira abies-marina and Fucus spiralis, Phaeophyta; Ulva compressa, Chlorophyta) were determined using the Kjeldahl method and the Weende method, respectively. The protein content ranged from 6.81 to 26.62 of dry weight for C. abies-marina and U. compressa, respectively. Fiber content was generally higher as compared with that in seaweeds from other origins and ranged from 33.82 to 63.88 for O. pinnatifida and F. spiralis, respectively

Antioxidant activity of macroalgae from the Azores

Free radical scavenging activity (FRSA) was studied to determine the antioxidant activity of 8 common macroalgae species found in the Azores. Species under study were Porphyra sp., Osmundea pinnatifida, Pterocladiella capillacea, Sphaerococcus coronopifolius and Gelidium microdon, from Rhodophyta; Ulva compressa and Chaetomorpha pachynema, from Chlorophyta and Fucus spiralis from Phaeophyta.The antioxidant activity was evaluated in methanolic extract by a decolourisation solution test of 2,2-diphenyl-1-picrylhydrazyl (DPPH), used as a stable radical. The methanolic extracts were obtained from dried biomass by sequential extractions, attaining a final concentration of 2 mg.mL-1. The FRSA values ranged from 19.54% for S. coronopifolius to 60.05% for F. spiralis with standard deviation (SD) varying between 1.41% and 6.80%. Results indicated that the studied seaweeds are a very promising source of biological active compounds with antioxidante properties. The seaweeds were collected in the Azorean islands where seawater pollution levels are low. Consequently, these seaweeds represent a valuable and good source of antioxidante material with superior beneficial effects on human health

Biotechnological applications of selected macroalgae

IBBA Strategic Planning - Workshop III, Plant Biotechnology Ponta Delgada, 25 Janeiro, 2011

Caracterização do sistema antioxidativo no estabelecimento da associação micorrízica Castanea sativa Mill. / Amanita muscaria

Poster apresentado no 5º Congresso Florestal Nacional, Viseu, Portugal, 16 Maio - 19 Maio 2005.As micorrizas são associações mutualistas que se estabelecem entre fungos, pertencentes sobretudo às divisões Basidiomycota, Ascomycota e Zigomycota, e raízes de plantas vasculares. Com o estabelecimento da associação micorrízica, ocorrem várias alterações morfológicas, fisiológicas e ecológicas em ambos os intervenientes, que têm sido objecto de estudo por diversos autores. Contudo, os mecanismos que controlam o processo de colonização e como este processo se inicia é completamente desconhecido. Os poucos trabalhos desenvolvidos nesta área, utilizando micorrizas arbusculares, sugerem que, durante as primeiras horas de invasão do fungo, ocorre uma indução de resposta de defesa por parte da planta hospedeira, semelhante à observada na interacção planta-patogénio. O presente trabalho pretende averiguar o efeito da inoculação de plantas de Castanea sativa Mill. com o fungo ectomicorrízico Amanita muscaria, na indução de resposta de defesa da planta hospedeira. O trabalho experimental decorreu num sistema in vitro estabelecido entre plantas de C. sativa e o fungo A. muscaria. Durante as primeiras horas de contacto (0 às 48 horas) procedeu-se à recolha de amostras de raízes, caules e folhas da planta hospedeira e de micélio de fungo que esteve em contacto com a raiz. Nestas amostras analisaram-se os níveis de peróxido de hidrogénio (H2O2), a actividade da catalase (CAT) (EC 1.11.1.6) e da superóxido dismutase (SOD) (EC 1.15.1.1). Os resultados obtidos evidenciam três picos de produção de H2O2, sugerindo o seu possível envolvimento no estabelecimento de micorrizas, nas primeiras horas de contacto planta-fungo. As variações observadas na actividade da SOD e da CAT, indiciam o seu envolvimento no controlo dos níveis de H2O2 em raízes, permitindo que esta molécula possa actuar como molécula sinalizadora, mas evitando que os níveis se tornem nocivos para o sistema. Estes resultados, apesar de preliminares, reflectem uma condição de stresse oxidativo semelhante àquela observada em interacções planta-patogénio.Fundação para a Ciência e a Tecnologia (FCT) - Programa Operacional “Ciência, Tecnologia, Inovação” (POCTI) - POCTI/BSE/38059/2001 - Study of the mechanisms regulating the interactions between ectomycorrhizal fungi and roots of Castanea sativa Mill. (European chestnut)