Browsing by Author "Figueiredo, A."
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- An apoplastic fluid extraction method for the characterization of grapevine leaves proteome and metabolome from a single samplePublication . Figueiredo, J.; Cavaco, A.R.; Guerra-Guimarães, L.; Leclerq, C.; Renaut, J.; Cunha, J.; Eiras-Dias, J.; Cordeiro, C.; Matos, A.R.; Silva, M.S.; Figueiredo, A.The analysis of complex biological systems keeps challenging researchers. The main goal of systems biology is to decipher interactions within cells, by integrating datasets from large scale analytical approaches including transcriptomics, proteomics and metabolomics andmore specialized ‘OMICS’ such as epigenomics and lipidomics. Studying different cellular compartments allows a broader understanding of cell dynamics. Plant apoplast, the cellular compartment external to the plasma membrane including the cell wall, is particularly demanding to analyze. Despite our knowledge on apoplast involvement on several processes from cell growth to stress responses, its dynamics is still poorly known due to the lack of efficient extraction processes adequate to each plant system.Analyzing woody plants such as grapevine raises even more challenges. Grapevine is among the most important fruit crops worldwide and awider characterization of its apoplast is essential for a deeper understanding of its physiology and cellular mechanisms. Here, we describe, for the first time, a vacuum-infiltrationcentrifugationmethod that allows a simultaneous extraction of grapevine apoplastic proteins and metabolites from leaves on a single sample, compatible with high-throughput mass spectrometry analyses. The extracted apoplast from two grapevine cultivars, Vitis vinifera cv ‘Trincadeira’ and ‘Regent’, was directly used for proteomics and metabolomics analysis. The proteome was analyzed by nanoLC-MS/MS and more than 700 common proteinswere identified, with highly diverse biological functions. The metabolome profile through FT-ICR-MS allowed the identification of 514 unique putative compounds revealing a broad spectrum of molecular classes
- Cephalopod fauna of the Pacific Southern Ocean using Antarctic toothfish (Dissostichus mawsoni) as biological samplers and fisheries bycatch specimensPublication . Queirós, J.P.; Ramos, J.A.; Cherel, Y.; Franzitta, M.; Duarte, B.; Rosa, R.; Monteiro, F.; Figueiredo, A.; Strugnel, J.M.; Fukuda, Y.; Stevens, D.W.; Xavier, J.C.Cephalopods are an important component of Southern Ocean food webs but studies analysing their habitat and trophic ecology are scarce. Here, we use the Antarctic toothfish Dissostichus mawsoni as a biological sampler of Southern Ocean’s cephalopods in the Ross, Amundsen, and D’Urville Seas. Ten cephalopod taxa were identified in the diet of the Antarctic toothfish, with Pareledone turqueti and Moroteuthopsis longimana being the only species present in all the three studied areas. DNA analysis conducted on squid flesh samples allowed identification of eight and two specimens of Mesonychoteuthis hamiltoni and M. longimana, respectively, proving this technique as a potential tool to improve the knowledge of cephalopods biodiversity and biogeography in the Southern Ocean. Stable isotopes were used to compare the habitat (δ13C) and trophic ecology (δ15N) between two life-stages of the two most abundant squid species (M. longimana and Psychroteuthis glacialis) from the D’Urville Sea (both squid species) and Amundsen Sea (only P. glacialis). Higher δ13C values in M. longimana suggest that this species inhabits waters near the Antarctic Polar Front, with incursions into sub-Antarctic waters, whilst P. glacialis spends its entire life in Antarctic waters. The most recently deposited part of the beak is enriched in 15N suggesting an increase in trophic level during squid growth. These results give us the first insights into the bathyal distribution of cephalopods in the Amundsen and D’Urville Seas, as well as into the ontogenetic changes of two of the most consumed squid species by top predators in this region. Such results are an important step towards improving the biogeography of Antarctic cephalopods, being of utmost importance to understand the biodiversity, food web structure, and functioning of this region
- Characterization of a Rat Model of Moderate Chronic Renal Failure-Focus on Hematological, Biochemical, and Cardio-Renal ProfilesPublication . Garrido, P.; Reis, F.; Costa, E.; Teixeira-Lemos, E.; Parada, B.; Alves, R.; Piloto, N.; Sereno, J.; Figueiredo, A.; Pinto, R.; Carvalho, L.; Rocha-Pereira, P.; Belo, L.; Santos-Silva, A.; Teixeira, F.The pathophysiological modifications underlying chronic renal failure seems to be dependent on the insufficiency degree, which will determine the moment to start therapy. As there is yet limited information about animal models of moderate chronic renal failure, we intended to perform a complete characterization of the hematological and cardio-renal alterations induced by partial nephrectomy. Blood samples from control and chronic renal failure rats were collected at 0, 3, 9, and 15 weeks in order to evaluate renal function, hematological parameters, iron metabolism, blood lipids, peripheral sympathetic nervous system, and inflammatory and redox status markers. BP, tissues trophy indexes, and kidney histomorphology were also assessed. Our data are consistent with a sustained moderate degree of chronic renal failure with a quickly compensated modest anaemia, though presenting iron metabolism disturbances. Despite the reasonable degree of functionality of the remnant kidney, as suggested by the anaemia correction and by the kidney hypertrophy and moderate lesions, several important cardiovascular modifications were developed. Our model presented hypertension, dyslipidemia, erythropoietic disturbances, sympathetic activation, and oxidative stress. This model might be a good tool to study the cellular/molecular mechanisms underlying moderate stages of chronic renal failure and to evaluate the therapeutic efficacy for prevention and treatment/correction of cardio-renal anaemia syndromes and complications in early stages.
- First steps on the resistance profiling of Kawisari coffee hybrid through cytological and gene expression analysesPublication . Diniz, I.; Figueiredo, A.; Sebastiana, M.; Munoz-Pajares, A.J.; Valverde, J.; Azevedo, H.; Rodrigues, A.S.; Prakash, R.S.; Pereira, A.P.; Guerra-Guimarães, L.; Azinheira, H.; Várzea, V.; Batista, D.; Silva, M.C.
- INHIBITION OF BLADDER TUMOR GROWTH BY A CYCLOOXYGENASE-2 INHIBITOR AND IMMUNOSUPPRESSION AGENTS IN AN EXPERIMENTAL STUDYPublication . Parada, B.; Reis, F.; de Lemos, Teixeira E.; Sereno, J.; Piloto, N.; Cunha, M.; Rocha-Pereira, P.; Pinto, R.; Mota, A.; Figueiredo, A.; Teixeira, F.
- Isolation of Apoplastic Fluid from Woody Plant Leaves: Grapevine and Coffee as a Case StudyPublication . Figueiredo, A.; Guerra-Guimarães, L.Proteomics is one of the key approaches to understand plant cell physiology involving the regulation of expression of many genes and metabolite production. Technical advances allowed a deeper characterization of plant proteomes, highlighting the need to study cellular compartments. The apoplast is the cellular compartment external to the plasma membrane including the cell wall, where a broad range of processes take place including intercellular signaling, metabolite transport, and plant–microbe interactions. Due to the fragile nature of leaf tissues, it is a challenge to obtain apoplastic fluids from leaves while maintaining cell integrity, which is particularly true for woody plants. Here, we describe the vacuum infiltrationcentrifugation (VIC) method for the extraction of the apoplastic fluid compatible with high-throughput proteomic approaches and biochemical analysis from different woody plants
- Metabolomics and transcriptomics to decipher molecular mechanisms underlying ectomycorrhizal root colonization of an oak treePublication . Sebastiana, M.; Gargallo-Garriga, A.; Sardans, J.; Pérez-Trujillo, M.; Monteiro, Filipa; Figueiredo, A.; Maia, M.; Nascimento, R.; Silva, M. Sousa; Ferreira, A. N.; Cordeiro, C.; Marques, A. P.; Sousa, L.; Malhó, R.; Peñuelas, J.Mycorrhizas are known to have a positive impact on plant growth and ability to resist major biotic and abiotic stresses. However, the metabolic alterations underlying mycorrhizal symbiosis are still understudied. By using metabolomics and transcriptomics approaches, cork oak roots colonized by the ectomycorrhizal fungus Pisolithus tinctorius were compared with non-colonized roots. Results show that compounds putatively corresponding to carbohydrates, organic acids, tannins, long-chain fatty acids and monoacylglycerols, were depleted in ectomycorrhizal cork oak colonized roots. Conversely, non-proteogenic amino acids, such as gamma-aminobutyric acid (GABA), and several putative defense-related compounds, including oxylipin-family compounds, terpenoids and B6 vitamers were induced in mycorrhizal roots. Transcriptomic analysis suggests the involvement of GABA in ectomycorrhizal symbiosis through increased synthesis and inhibition of degradation in mycorrhizal roots. Results from this global metabolomics analysis suggest decreases in root metabolites which are common components of exudates, and in compounds related to root external protective layers which could facilitate plant-fungal contact and enhance symbiosis. Root metabolic pathways involved in defense against stress were induced in ectomycorrhizal roots that could be involved in a plant mechanism to avoid uncontrolled growth of the fungal symbiont in the root apoplast. Several of the identified symbiosis-specific metabolites, such as GABA, may help to understand how ectomycorrhizal fungi such as P. tinctorius benefit their host plants.
- Modulation of apoplast proteome by downy mildew in susceptible and tolerant grapevine cultivarsPublication . Figueiredo, Joana; Cavaco, A.R.; Guerra-Guimarães, L.; Leclerq, C.C.; Santos, R.B.; Silva, M.C.; Renaut, J.; Sousa Silva, M.; Figueiredo, A.
- O Projecto INTERFRUTA II e o estudo dos problemas fitossanitários das fruteiras dos AçoresPublication . Mexia, A.M.M.; Lopes, D.H.; Raimundo Perez, C.; Aguin-Pombo, D.; Dantas, L.; Pereira, A.; Rodrigues, R.; Figueiredo, A.; Carvalho, M.C.; Silva, M.L.; Mumford, J.The INTERFRUTA project is financed by the European Commission Interreg III-B Programme and was developed for the islands of “Madeira”, “Tenerife” and “Terceira”, for the improvement of fruit and vineyard production in these three Atlantic regions. The project goal is a better knowledge of the phytossanitary problems that affect apples, bananas, chestnut and vineyards, applying methods that will contribute to solutions based on the survey of key pests, diseases and beneficial organisms. In the three fruit crops and vineyards analyzed, the methodology used was mainly direct observation of predefined parts of each plant (such as leaves and flowers) on which pests might be expected to occur. The key pest C. capitata was monitored as adults using sexual lures in a net of traps around the island of “Terceira” up to 200 meters altitude. The beneficial insect survey was conducted using the beating technique. For diseases, a direct sampling was carried out by the observation of 10 % of the plants in each orchard or vineyard plot looking for visible symptoms. In apples and vineyards, flowers and branches were collected and tested for microorganisms using serologic and molecular techniques (Elisa and PCR). In banana plantations, the most damaging pests were banana weevil, thrips and a fungi complex that causes symptoms similar to Fusarium wilt. In chestnut, the pest that causes the most severe damage, affecting around 40 % of the nuts produced, is the Chestnut tortrix, but there were some difficulties in identifying which of the three known species was present in the Azores islands. In vineyards, diseases that are most frequent and important include grapevine downy mildew, powdery mildew and Botrytis. Invertebrate pests included grape leafhoppers, thrips and eriophyid mites. The European vine moth was also surveyed but none were found and is still not present. In apple orchards the most important problems were thrips, mites, oriental fruit moth and codling moth. Anarsia lineatella Zeller, the peach twig borer, was not detected and new Eriophyid species have been recorded for the first time in the Azores as a result of the surveys. With the information collected it was possible to create a phytossanitary diagnostic database for the Macaronesian islands (PROFITOMAC) on the project internet pages (www.interfruta.uac.pt), to help users to identify problems that affect the studied cultures.
- Proteomic signatures uncover the early key players on Vitis vinifera cv. “Regent - Plasmopara viticola crosstalkPublication . Figueiredo, Joana; Santos, Rita B.; Cavaco, A.R.; Guerra-Guimarães, Leonor; Leclerq, C.C.; Sousa, L.; Silva, M.C.; Renaut, J.; Silva, M.S.; Figueiredo, A.