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- Identification and validation of microsatellite markers in strawberry tree (Arbutusunedo L.)Publication . Fazenda, Pedro; Pereira, Ricardo; Fonseca, Maria; Carlier, Jorge; Leitão, JoséStrawberry tree (Arbutus unedo L.), an evergreen shrub/small tree of the family Ericaceae, is a main constituent of the Mediterranean basin flora; although it is also found in southwestern Prance, Macaronesia, and Ireland. The small fruits are edible but mostly used for preparation of preserves and jams, and for liquors such as the Portuguese traditional "aguardente de medronho". Traditionally cultivated by small farmers, often in consociation with Quercus sp., strawberry tree is presently emerging as a new important fruit crop cultivated in large orchards by modern export-oriented enterprises. This change of paradigm requires a growing role of plant breeding, upstream of the production process. Genomic tools for this species are mostly limited to the chloroplast genome sequence and to genomic data described in this work. In order to identify strawberry tree microsatellite (SSR) loci we performed partial genome next-generation sequencing using the Ion Torrent technology. The sequenced similar to 24.6M nucleotides resulted in the identification of 1185 microsatellite markers mostly constituted by dinucleotide motifs. The relative amount of microsatellite dinucleotide motifs (AG/CT - 71.7%, AC/GT - 20.5%, AT/AT - 2.9%, and CG/CG - 0.3%) is similar to the one observed in other Ericaceae species. Among a tested sample of 40 SSR primer pairs, 20 amplified well-defined PCR products, 12 (30%) were validated as polymorphic. Used in our collaborative project for molecular identification of selected and improved clones, the identified SSR loci constitute a strong tool for a large panoply of applied and fundamental studies of this emerging fruit crop.
- Feasibility of co-treating olive mill wastewater and acid mine drainagePublication . Carlier, Jorge; Luís, Ana; Alexandre, Luís Miguel; Costa, Maria ClaraPrevious tests using a growth medium and olive mill wastewater (OMWW) have shown that it supplies carbon and electron donors suitable for sulphate reducing bacteria (SRB). We assessed the co-treatment of acid mine drainage (AMD) and OMWW using SRB-enriched bioreactors and identified the most abundant bacterial populations present under optimized conditions. The process requires a neutralizing agent to create optimal pH conditions for successful removal of the AMD's main contaminants. Concentrations of SO42-, Al, Fe, Cu, Zn, and Mn decreased to below Portugal's maximum admissible values for irrigation waters, and all but Mn were reduced to less than Portugal's emission limit values (ELVs) for wastewater discharges. Phenol concentrations-the main pollutants in OMWW-dropped to values between 1/10 and 1/5 their initial concentrations in batch tests using mixtures of AMD and OMWW, and to 1/2 their initial concentrations in flow-through tests. The final total phenol concentrations were still above the ELV for wastewater discharges, but phenols are not regulated in irrigation waters, and OMWW is used by some producers to irrigate soils. Six main SRB groups were identified as likely having a fundamental role in the bioremediation process: the generaDesulfovibrio,Sulfurospirillum, andAcetobacterand the familiesSphingomonadaceae,Prevotellaceae, andDeferribacteraceae
- Physicochemical characterization and antioxidant activity of honey with Eragrostis spp. pollen predominancePublication . AAZZA, Smail; Elamine, Youssef; EL-GEUNDOUZ, Soukaina; Lyoussi, Badiaa; Antunes, Maria Dulce; Estevinho, Leticia M.; Anjos, Ofélia; Carlier, Jorge; Costa, Maria C.; Miguel, MariaThis study aimed to characterize the pollen and physicochemical profiles, and to evaluate the antioxidant capacity of six samples of Moroccan honey. The pollen grains of Eragrostis spp. were predominant in all samples (from a minimum of 46.581.59% to 72.72 +/- 2.43% as a maximum), exceeding the minimum (generally 45%) required for the monoflorality nomination of a honey sample, and suggestion a case of newly reported honey. The analyzed samples had low diastase activity and hydroxymethylfurfural (HMF) content. Potassium was the dominant element in practically all samples, except the sample with the highest Eragrostis spp. pollen percentage, which showed sodium predominance. The honey color ranged from water white to light amber, and positively correlated the concentrations of phenols, proline and the capacity for scavenging free radicals. However, great variability observed in all the analyzed parameters, and could not be explained by the variability of the Eragrostis spp. pollen percentage, which suggests a hypothesis of low concentrated nectar. Practical applicationsThe melissopalynological profile of honey is an important parameter for determining the type of honey (multifloral, monofloral, blossom, honeydew) and its relationship to the botanical and geographical provenance. There is generally a relationship between the pollen profile of honey and the biological properties. The physicochemical characterization, antioxidant activity and melissopalynological profile of Eragrostis honey was reported for the first time in this study. These data intend to contribute for the introduction of a new type of honey labeled as Eragrostis honey. At the same time, the evaluation of the antioxidant activity may also trigger the search of other biological properties with medicinal attributes for the new Eragrostis ssp. honey.
- Prokaryotic diversity in stream sediments affected by acid mine drainagePublication . Carlier, Jorge; Ettamimi, Sara; Cox, Cymon J.; Hammani, Khalil; Ghazal, Hassan; Costa, Maria ClaraThe microbial communities in mining impacted areas rely on a variety of mechanisms to survive in such extreme environments. In this work, a meta-taxonomic approach using 16S rRNA gene sequences was used to investigate the prokaryotic diversity of sediment samples from water bodies affected by acid mine drainage at the Sao Domingos mining area in the south of Portugal. Samples were collected in summer and winter from the most contaminated sites from where the water flows downstream to the freshwater of Chanca's river reservoir. The prokaryotic diversity on water bodies' sediments allowed us to distinguish the highly contaminated sites (pH approximate to 2) from sites with intermediate levels of contamination (pH approximate to 3-6.5), and from sites without contamination (pH approximate to 7.5). The abundances of acidophiles of generaAcidiphilium, Acidibacter, Acidobacterium and Acidocellain the sediments were correlated with the level of acid mine drainage contamination. The two first genera were among the 30 most abundant prokaryotes in all contaminated samples, including one (SS2w), where the contamination was very diluted, thereby emphasizing the impact that such type of pollution can have in the microbial communities of sediments. In addition, the high abundances of archaeal taxa from classThermoplasmataand of bacteria from family RCP1-48 in the sediments from the most contaminated site corroborate their importance in such ecosystems and a putative role in the generation of acid mine drainage.
- Main results from the EU-funded project "evaluation and utilization of pineapple genetic resources from the amazon to breed resistant varieties"Publication . D'Eeckenbrugge, G. Coppens; Cabral, J. R. S.; Matos, A. P. de; Carlier, Jorge; Leitão, José; Duval, M. F.; Noyer, J. L.; Ferreira, F. R.; Leal, F.; Maggioni, L.; Suarez, Z.This project was aimed at obtaining knowledge and tools for the development of cultivars resistant to the main diseases in South America, by (i) characterizing and evaluating available genetic resources, (ii) studying the genetic structure of the genera Ananas and Pseudanas (iii) developing and applying resistance screening techniques for important pests and disease in the region, (iv) studying the heredity of agronomic traits, and (v) testing the potential of partial inbreeds in breeding. In addition, a standard database is being established in collaboration with IPGRI, to promote information on germplasm exchange. The project, which started in 1997 and ended in 2001, has produced an inventory of the germplasm held in its collections, allowing exchange and repatriation of lost germplasm. The list of descriptors has been revised. Multivariate analyses of the morphological data have produced consistent results. Molecular characterization, based on RFLP of genomic DNA and PCR-RFLP of cpDNA has confirmed the proximity of Ananas and Pseudananas, the absence of interspecific barriers in Ananas, the importance of a geographic component of variation, and the existence of a higher genetic diversity in the North of South America. Pineapple taxonomy has been revised. Genetic mapping was based on isozyme, RAPD, AFLP and ISSR markers and was carried out on a hybrid progeny between A. bracteatus and A. comosus, following the pseudotestcross approach. It resulted in a first map of 390 and 174 DNA markers distributed in 46 and 31 linkage groups for these two species. New sources of resistance to fusariosis have been identified, some of which show other interesting traits for the Brazilian producers. According to the first inheritance studies, this resistance seems monogenic and dominant, a result still to be confirmed. Selfprogenies obtained from the cultivars Perolera, Primavera, and Roxo de Tefé, showed inbreeding depression, but normal segregation for major leaf traits (spininess, colour). Subsequent hybridization of the inbreeds restored vigour.
- Design of remediation pilot plants for the treatment of industrial metal-bearing effluents (BIOMETAL DEMO project): Lab testsPublication . Ballester, Antonio; Castro, Laura; Clara Costa, Maria; Carlier, Jorge; Garcia-Roig, Manuel; Perez-Galende, Patricia; Alvarez, Angela; Bertagnolli, Caroline; Guibal, EricCurrent research is the first part of the BIOMETAL DEMO project, funded by the European Union, focusing on the development of complementary and modular biotreatment processes for the removal of metal ions from industrial effluents (issued from mining industry, surface treatment and ceramics). Metal bio-precipitation (in form of metal sulfides and metal phosphates) using sulfate-reducing bacteria (SRB), phytase and phosphatase enzymes, biosorption using sugar-beet pulp, algal biomass and alginate- or chitosan-based composites (immobilizing polyethyleneimine derivatives by encapsulation) have been extensively studied using synthetic and industrial effluents. These preliminary results allow designing a treatment flow-sheet including pre-treatment of the effluent using biologically-assisted precipitation followed by biosorption (for mining effluents that contain sulfate anions). For surface treatment and ceramics effluents, bio-precipitation combined with biosorption processes and physicochemical pretreatment followed by biosorption process represent, respectively, a promising solution. The carbon-source for the SRB process, the optimization of process parameters and the biosorbents have been selected taking into account both the technical performance and the economic viability. (C) 2016 Elsevier B.V. All rights reserved.
- Molecular characterization of Phaseolus vulgaris L. genotypes included in Bulgarian collection by ISSR and AFLP (TM) analysesPublication . Svetleva, D.; Pereira, G.; Carlier, Jorge; Cabrita, L.; Leitão, J. M.; Genchev, D.Seventy-eight (33 Bulgarian and 45 foreign) common bean (Phaseolus vulgaris L.) genotypes included in Bulgarian collection were screened for ISSR (Inter Simple Sequence Repeats) and AFLP (TM) (Amplified Fragment Length Polymorphism) markers. ISSR analysis was performed with 13 primers, and 55 (36.7%) out of the 150 bands observed were polymorphic. One hundred and sixty-four AFLP (TM) fragments were obtained with three primer combinations, of which 54 (32.9%) were polymorphic. UPGMA (Unweighted Pair Group Method Arithmetic Averages) analysis was performed using DICE coefficient and dendrograms were constructed using either the data from each method (ISSR and AFLP (TM)) separately or combined in a single joint matrix. Despite some genetic heterogeneity observed in both pools (Middle American origin: M and Andean origin: A) the genotypes were separated in to main groups: one gathering genotypes mainly from pool M and the other more genotypes from pool A. Moreover, Bulgarian genotypes were spread over the two groups suggesting that they are not subject to genetic erosion and that the Phaseolus genetic diversity is conserved. (c) 2006 Elsevier B.V. All rights reserved.
- Comparative study of the antioxidant and enzyme inhibitory activities of two types of Moroccan Euphorbia entire honey and their phenolic extractsPublication . Boutoub, Oumaima; EL-GUENDOUZ, Soukaina; Manhita, Ana; Dias, Cristina Barrocas; Estevinho, Letícia M.; Paula, Vanessa B.; Carlier, Jorge; Costa, Maria Clara; Rodrigues, Brígida; Raposo, Sara; Aazza, Smail; El Ghadraoui, Lahsen; Miguel, Maria GraçaHoney is a natural food product very famous for its health benefits for being an important source of antioxidant and phenolic compounds. Euphorbia honeys obtained from different regions of Morocco were evaluated for their ability to inhibit acetylcholinesterase, lipoxygenase, tyrosinase and xanthine oxidase activities. Their antioxidant properties were evaluated using the: 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging capacity, nitric oxide scavenging activity (NO) and scavenging ability of superoxide anion radical. Then, the phenolic extracts of the same entire honey samples were evaluated by liquid chromatography coupled to diode array detection and mass spectrometry (LC-DAD-MS) and tested for the biological activities previously evaluated on the entire honeys, in order to conduct a comparative study between both (honey and phenolic extracts). The chromatographic profiles for the studied Euphorbia honey extracts were different. Phenolic compounds gallic acid, 4-hydroxybenzoic acid and p-coumaric acid were detected in all samples, whereas kampferol was only present in two samples. Physicochemical parameters and total phenolic content were also determined. Entire honey that recorded the highest rate of phenols was sample M6 (E. resinifera) = 69.25 mg GAE/100 g. On the other hand, the phenolic extracts had better antioxidant and enzyme inhibitory activities than the entire honeys, regardless the monofloral honey type. In conclusion, the studied Euphorbia honeys may have a great potential as antioxidant, anti-inflammatory and anti-tyrosinase sources for pharmaceutical and cosmetic applications.
- Impact of lead (Pb2+) on the growth and biological activity of Serratia marcescens selected for wastewater treatment and identification of its zntR gene—a metal efflux regulatorPublication . dos Reis Ferreira, Gustavo Magno; Pires, Josiane Ferreira; Ribeiro, Luciana Silva; Dias Carlier, Jorge; Costa, Maria Clara; Schwan, Rosane Freitas; Silva, Cristina FerreiraMicroorganisms isolated from contaminated areas play an important role in bioremediation processes. They promote heavy metal removal from the environment by adsorbing ions onto the cell wall surface, accumulating them inside the cells, or reducing, complexing, or precipitating these substances in the environment. Microorganism-based bioremediation processes can be highly efcient, low-cost and have low environmental impact. Thus, the present study aimed to select Pb2+-resistant bacteria and evaluate the growth rate, biological activity, and the presence of genes associated with metal resistance. Serratia marcescens CCMA 1010, that was previously isolated from cofee processing wastewater, was selected since was able to growth in Pb2+ concentrations of up to 4.0 mM. The growth rate and generation time did not difer from those of the control (without Pb2+), although biological activity decreased in the frst hour of exposure to these ions and stabilized after this period. The presence of the zntR, zntA and pbrA genes was analysed, and only zntR was detected. The zntR gene encodes a protein responsible for regulating the production of ZntA, a transmembrane protein that facilitates Pb2+ extrusion out of the cell. S. marcescens CCMA 1010 demonstrated a potential for use as bioindicator that has potential to be used in bioremediation processes due to its resistance to high concentrations of Pb2+, ability to grow until 24 h of exposure, and possession of a gene that indicates the existence of mechanisms associated with resistance to lead (Pb2+).
- Leaf mineral element content and soil characteristics on in vitro antioxidant and enzymatic inhibitory activities of aqueous fennel extractsPublication . MAJDOUB, Nesrine; EL-GEUNDOUZ, Soukaina; Carlier, Jorge; Clara Costa, Maria; Guerrero, Carlos; Duarte, João; Miguel, Mariahis study was conducted to evaluate the biochemical characterization of three harvested Foeniculum vulgare plants collected from two bioclimatic zones in order to investigate the soil growing conditions effect. The results showed a great variability of the phenolic amounts and biological properties of samples rely on localities. FvSEN contained the highest amounts of phenolic compounds. These amounts were accompanied by the greatest antioxidant ability through almost studied assays. FvSEN and FvZO were significantly different. In addition, the samples exhibited a significant and variable enzymatic inhibition activity with values ranging from 30 to 50 mu g/mL for lipoxygenase assay. But these extracts did not revealed significant differences on their tyrosinase abilities. On the other hand, the levels of mineral elements were also estimated. These contents varied depending on sample and locality. The growing soil conditions of samples in terms of different parameters is likely related to their antioxidant and enzyme inhibition potentialities added to their mineral composition which settled by Spearman's correlation. These data may confirm the interesting potential of F. vulgare as a valuable source for natural antioxidant molecules but the growing soil conditions can affect all the potentialities of these plants set for human consumption and other uses.