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Holocentric chromosomes are rare among angiosperms, but have been suggested to be shared by all or most of the species of Cyperaceae and Juncaceae. However, no clear demonstration of the centromere type in Juncus, the largest genus of Juncaceae, has so far been published. Thus, we conducted a detailed chromosomal investigation of four Juncus spp. aiming to identify their centromere type. Mitotic chromosomes were analysed using the fluorochromes CMA and DAPI, fluorescent in situ hybridization (FISH) with rDNA probes and immunodetection of histones H3 phosphorylated at serine 10 (H3-S10ph) and H2A phosphorylated at threonine 133 (H2A-T133ph). DAPI-stained chromosomes of all species displayed typical primary constrictions, which were not related to AT-poor CMA+ heterochromatin or rDNA sites (usually negatively stained with DAPI). Immunodetection with H3-S10ph and H2A-T133ph revealed hyperphosphorylation of pericentromeric and centromeric regions, respectively, in a restricted area, as observed in monocentric chromosomes. Meiotic analyses in J. microcephalus showed no indication of inverted meiosis, commonly found in plants with holocentric chromosomes. Since the species investigated here belong to four different sections of Juncus and all of them display typical monocentric chromosomes, it seems that this kind of centromere is common in the genus and may represent the standard centromere organization for Juncus. If Juncus has monocentric chromosomes, there is no reason to hypothesize that other genera of Juncaceae for which centromeres have not been carefully investigated have holocentric chromosomes.

O objetivo deste estudo foi avaliar estádios de maturação do fruto e temperaturas na qualidade fisiológica de sementes de pimenta biquinho. Dois experimentos foram conduzidos, no primeiro, os frutos foram colhidos em diferentes estádios de maturação: frutos de coloração verde, alaranjado, vermelho, vermelho intenso e vermelho intenso e murchos. Das sementes extraídas determinou-se o teor de água e avaliou-se a qualidade fisiológica por meio dos testes de germinação e vigor (primeira contagem de germinação, emergência de plântulas, índice de velocidade de emergência, comprimento, matéria fresca e seca de plântulas). No segundo experimento, as sementes provenientes de frutos vermelhos e vermelho intenso e murchos foram submetidas a diferentes condições de temperaturas: constante (20, 25 e 30 °C) e alternada (20-30 °C), e novamente avaliadas pelos testes de germinação e vigor. A qualidade fisiológica das sementes de pimenta biquinho é máxima quando os frutos são colhidos com a coloração vermelho intenso e/ou vermelho intenso e murcho e, as temperaturas constantes de 25 e 30 °C são as mais adequadas para avaliação da qualidade fisiológica de sementes.

Objetivou-se avaliar a qualidade fisiológica de sementes de crambe tratadas com fertilizante a base de zinco e molibdênio. O experimento foi conduzido no Laboratório de Análise de Sementes da Unimontes. O delineamento experimental utilizado foi inteiramente casualizado, com quatro repetições. Os tratamentos testados constituíram de cinco doses de fertilizante comercial composto por zinco (35%) e molibdênio (3,5%), aplicados via semente nas concentrações de 0; 3,3; 6,7; 10 e 13 mL do fertilizante para 0,5 Kg-1 de sementes. A qualidade fisiológica das sementes foi avaliada por meio dos seguintes testes: germinação, primeira contagem de germinação, emergência de plântulas e comprimento de plântulas. Com o aumento das doses do fertilizante comercial composto por zinco e molibdênio, houve redução na germinação e vigor das sementes de crambe. A qualidade fisiológica das sementes de crambe é influenciada negativamente por doses crescentes de molibdênio.

This study aimed to evaluate the effect of maturation fruit and drying temperatures stages on oil content and the physiological performance of crambe seeds. Randomized blocks were the experimental design, with four replications. Harvests occurred when the plants reached 20, 40, 60, 80 and 100% brown color fruits. After harvest, seeds were dried at different temperatures (natural and artificial air at 30, 45 and 60 °C). The seeds were evaluated for the weight of a thousand seeds, productivity, oil content, germination and vigor (count of the first germination and seedling emergence). Crambe seeds, FMS Brilhante cultivar, reach the maximum oil content with harvests made from 70% of total brown color fruits. Drying temperatures do not affect the oil content of the seeds. Harvesting crambe seed aiming at the highest physiological quality should occur when the plants have between 75 and 85% brown fruits. Artificial drying at 30 and 45 °C can be recommended for crambe seeds, favoring the production of quality seeds. Resumo: Este estudo teve como objetivo avaliar o efeito dos estádios de maturação dos frutos e das temperaturas de secagem sobre o teor de óleo e o potencial fisiológico das sementes de crambe. O delineamento experimental foi o de blocos casualizados, com quatro repetições. A colheita ocorreu quando as plantas atingiram 20, 40, 60, 80 e 100% de frutos de cor marrom. Após a colheita, as sementes foram secas em diferentes temperaturas (ar natural e artificial a 30, 45 e 60 °C). As sementes foram avaliadas quanto ao peso de mil sementes, produtividade, teor de óleo, germinação e vigor (primeira contagem de germinação e emergência de plântulas). As sementes de crambe, cultivar FMS Brilhante, atingiram o teor máximo de óleo com colheitas feitas a partir de 70% do total de frutos de cor marrom. As temperaturas de secagem não afetaram o teor de óleo das sementes. A colheita de sementes de crambe visando a mais alta qualidade fisiológica deve ocorrer quando as plantas tiverem entre 75 e 85% de frutos marrons. A secagem artificial a 30 e 45 °C pode ser recomendada para sementes de crambe, favorecendo a produção de sementes de qualidade.

Natural compounds have had increasing applications in the biotechnological sector, with a large fraction of these substances being channeled to the pharmaceutical industry due to their important pharmacological properties. The discovery of new bioactive molecules with novel mechanisms of action constitutes a promising solution for the design of alternative therapeutic solutions. Actinobacteria are a large group of morphologically and physiologically diverse bacteria well known for their production of biotechnologically relevant compounds. The Portuguese coast is scantly explored in terms of Actinobacteria diversity and respective bioactive potential, offering a good opportunity to find new Actinobacteria taxa and bioactive natural products. In this study, we investigated the Actinobacteria diversity associated with a sediment sample collected from the intertidal zone of a beach in northern Portugal, through a cultivation-dependent approach, and screened its antimicrobial and cytotoxic potential. A total of 52 Actinobacteria strains were recovered from the marine sediment, with the largest fraction of the isolates belonging to the genus Micromonospora. Bioactivity screening assays identified crude extracts of six Streptomyces strains active against C. albicans, exhibiting minimum inhibition concentration (MIC) values in the range of 3.90&ndash, 125 &mu, g mL&minus, 1. Twenty-five Actinobacteria crude extracts (obtained from strains of the genera Micromonospora, Streptomyces and Actinomadura) exhibited significant effects on the viability of at least one tested cancer cell line (breast ductal carcinoma T-47D and liver hepatocellular carcinoma HepG2). The Actinobacteria extracts demonstrating activity in the antimicrobial and/or cytotoxic assays were subjected to metabolomic analysis (Mass spectrometry (MS)-based dereplication and molecular networking analyses), indicating the presence of four clusters that may represent new natural products. The results obtained demonstrate the importance of bioprospecting underexplored environments, like the Portuguese coast, for enhancing the discovery of new natural products, and call attention to the relevance of preserving the natural genetic diversity of coastal environments.

A produção de sementes é um processo que incluem diversas etapas desde o melhoramento, produção, certificação, até a manutenção depois da colheita, cuja finalidade especifica é obter sementes de qualidade, com alta pureza genética, pureza física, qualidade fisiológica e sanitária, que afetam a sua capacidade de originar plantas de alta produtividade. Objetivou-se com esta revisão abordar os principais aspectos práticos verificados durante a produção de sementes de crambe, visando obter sementes de elevada qualidade física, fisiológica, genética e sanitária. A metodologia estudada foi a revisão de literatura com levantando e análise de dados sobre a cultura. Verificou-se que para sementes de crambe ainda não há na literatura informações suficientes quanto às diversas etapas de um sistema de produção de sementes, especialmente no que se refere ao estabelecimento e condução de campos de produção. Diante destas considerações, o conhecimento das condições ideais durante a produção de sementes é de fundamental importância, uma vez que somente aquelas de elevado nível de qualidade proporcionam a maximização da ação dos demais insumos e fatores de produção empregados na lavoura, com respostas na produtividade das culturas.

In the search for more efficient techniques for the propagation of peach cuttings, this study aimed to evaluate the rooting of hardwood cuttings of peach rootstock genotypes under different indolebutyric acid (IBA) concentrations. In the winter of 2016, the basal end of cuttings of genotypes 1701-1, 1701-2, 102-1, 102-2, 202-1 and 'Okinawa' were dipped into solutions with five concentrations of IBA (0, 1000, 2000, 3000 and 4000 mg mg L-1) for 5 seconds. The cuttings were then placed in sand in plastic trays and kept in a mist chamber. The experiment was arranged in a 6x5 factorial, completely randomized design, with 5 replications, and each plot consisted of eight cuttings. After 59 days, the variables related to rooting and root quality were evaluated. Genotypes 102-1 and 202-1 showed high adventitious rooting potential in hardwood cuttings, with 76.8 and 66.5% of rooting, respectively. The concentration of 2000 mg.L-1 of IBA acid can be recommended for the treatment of hardwood cuttings of the tested rootstocks for propagation in the winter. Buscando técnicas mais eficientes na propagação de mudas de pessegueiro, desenvolveu-se este trabalho com o objetivo de avaliar o enraizamento de estacas lenhosas de genótipos de porta-enxerto de pessegueiro submetidos a diferentes concentrações de ácido indolbutírico (AIB). No inverno de 2016, as estacas dos genótipos 1701-1, 1701-2, 102-1, 102-2, 202-1 e do porta-enxerto ‘Okinawa’ foram tratadas por imersão da base por cinco segundos em cinco doses de AIB (0, 1.000, 2.000, 3.000 e 4.000 mg L-1). A estaquia foi realizada em caixas plásticas contendo areia, em câmara de nebulização. O delineamento experimental foi o inteiramente casualizado, com 5 repetições em arranjo fatorial 6x5, sendo cada parcela composta por oito estacas. Transcorridos 59 após implantação do experimento, foram avaliadas variáveis relativas ao enraizamento e à qualidade das raízes. Os genótipos 102-1 e 202-1 apresentam alto potencial de enraizamento adventício em estacas lenhosas, com enraizamento de 76,8 e 66,5%, respectivamente. O ácido indolbutírico, na concentração de 2.000 mg.L-1, pode ser recomendado para o tratamento de estacas lenhosas dos porta-enxertos testados, para estaquia no inverno.

The seed treatment provides disease control before the crop installation, minimizing the pathogens control cost. However, there are few seed treatment alternatives to organic farming. In this sense, the objective of the study was to evaluate the physiological and sanitary quality of organic tomato seeds treated with clove basil extract. The experiment was carried out in a completely randomized design with a 2x5 factorial (extract concentrations x imbibition periods) for each seed lot. Six lots of tomato seeds produced under organic cultivation were used. The samples of the lots were stored in glass pots in a cold room at 16 ºC and 80% of relative humidity for 12 months. Seeds were treated with clove basil extract at concentrations of 5 and 10% for 0; 2.5; 5; 7.5 and 10 minutes. The seeds physiological and health quality were evaluated by the following tests: moisture content, germination, first count of germination, seedling emergence, emergence speed index and seed health. In general, the wild basil extract does not affect the physiological quality of tomato seeds. There were inhibitory and antifungal activity of Aspergillus sp. using basil extract, significantly reducing their impact on the seeds, for all evaluated factors. It was concluded that the clove basil extract inhibit the Aspergillus sp. incidence in organic tomato seeds, maintaining its physiological quality.

A dormência é um fenômeno intrínseco da semente, funcionando como mecanismo de resistência natural aos fatores adversos do meio. A germinação só ocorrerá quando esta for superada e as condições ambientais forem favoráveis ao crescimento das plântulas. O objetivo deste trabalho foi analisar o efeito de tratamentos para superação da dormência na estrutura da testa de sementes de Apuleia leiocarpa pela microscopia eletrônica de varredura. Os métodos pré-germinativos para superação da dormência foram: sementes não escarificadas (testemunha), imersão em ácido sulfúrico 75 e 98% por três e nove minutos e imersão em água aquecida a 80ºC por 30 segundos. As sementes foram submetidas às análises em microscopia e foram avaliadas pelo teste de germinação. Os tratamentos foram eficientes em romper a exotesta das sementes de garapa, sendo que o ácido promoveu elevado incremento na germinação. A água aquecida causou a morte das sementes.

Water stress is a condition that causes physiological changes in different species and even genotypes of the same species. One of the osmotic agents most used to simulate this condition is polyethylene glycol 6000. The aim of this study was to evaluate the effects of water stress on the physiological quality of seeds and performance of seedlings of crambe genotypes. A completely randomized design was used, in a factorial scheme with two genotypes (one cultivar and one line) and five osmotic potentials simulated with aqueous solutions of polyethylene glycol 6000 (control = 0, -0.25, -0.50, -1.0, -1.50 MPa), with four replicates of 50 seeds per treatment. The effects of the treatments were evaluated by means of germination, first count, germination speed index and fresh and dry matter of seedlings. The physiological quality of seeds and the performance of crambe seedlings are negatively affected under water stress from -0.25 MPa on. Seeds of the genotype FMSCR 1101 have greater tolerance to drought stress, regardless of the osmotic potential used.

We demonstrated by cytogenomic analysis that the proximal heterochromatin of the Northeast Brazilian species of Caesalpinia group is enriched with phylogenetically conserved Ty3/Gypsy-Tekay RT, but diverge in the presence of Ty3/Gypsy-Athila RT and satDNA. The Caesalpinia Group includes 225 species and 27 monophyletic genera of which four occur in Northeastern Brazil: Erythrostemon (1 sp.), Cenostigma (7 spp.), Libidibia (1 sp.), and Paubrasilia (1 sp.). The last three genera are placed in different clades in the Caesalpinia Group phylogeny, and yet they are characterized by having a numerically stable karyotype 2n = 24 (16 M+8A) and GC-rich heterochromatic bands (chromomycin A3 positive/CMA+ bands) in the proximal chromosome regions. To characterize the composition of their heterochromatin and test for the homology of these chromosomal regions, genomic DNA was extracted from Cenostigma microphyllum, Libidibia ferrea, and Paubrasilia echinata, and sequenced at low coverage using the Illumina platform. The genomic repetitive fractions were characterized using a Galaxy/RepeatExplorer-Elixir platform. The most abundant elements of each genome were chromosomally located by fluorescent in situ hybridization (FISH) and compared to the CMA+ heterochromatin distribution. The repetitive fraction of the genomes of C. microphyllum, L. ferrea, and P. echinata were estimated to be 41.70%, 38.44%, and 72.51%, respectively. Ty3/Gypsy retrotransposons (RT), specifically the Tekay lineage, were the most abundant repeats in each of the three genomes. FISH mapping revealed species-specific patterns for the Tekay elements in the proximal regions of the chromosomes, co-localized with CMA+ bands. Other species-specific patterns were observed, e.g., for the Ty3/Gypsy RT Athila elements which were found in all the proximal heterochromatin of L. ferrea or restricted to the acrocentric chromosomes of C. microphyllum. This Athila labeling co-localized with satellite DNAs (satDNAs). Although the Caesalpinia Group diverged around 55 Mya, our results suggest an ancestral colonization of Tekay RT in the proximal heterochromatin. Thus, the present-day composition of the pericentromeric heterochromatin in these Northeast Brazilian species is a combination of the maintenance of an ancestral Tekay distribution with a species-specific accumulation of other repeats.

Repetitive sequences are ubiquitous and fast-evolving elements responsible for size variation and large-scale organization of plant genomes. Within tribe Phaseoleae (Fabaceae), some genera, such as Phaseolus, Vigna, and Cajanus, show small genome and mostly stable chromosome number. Here, we applied a combined computational and cytological approach to study the organization and diversification of repetitive elements in some species of these genera. Sequences were classified in terms of type and repetitiveness and the most abundant were mapped to chromosomes. We identified long terminal repeat (LTR) retrotransposons, especially Ogre and Chromovirus elements, making up most of genomes, other than P. acutifolius and Vigna species. Satellite DNAs (SatDNAs) were less representative, but highly diverse among species, showing a clear phylogenetic relationship. In situ localization revealed preferential location at pericentromeres and centromeres for both types of sequences, suggesting a heterogeneous composition, especially for centromeres. Few elements showed subterminal accumulation. Copy number variation among chromosomes within and among species was observed for all nine identified SatDNAs. Altogether, our data pointed two main elements (Ty3/Gypsy retrotransponsons and SatDNAs) to the diversification on the repetitive landscape in Phaseoleae, with a typical set of repeats in each species. The high turnover of these sequences, however, did not affect total genome size.

Nature is the major reservoir of biologically active molecules. The urgent need of finding novel molecules for pharmaceutical application is prompting the research of underexplored environments, such as marine ecosystems. Here, we investigated cultivable actinobacteria associated with the macroalgae Laminaria ochroleuca and assessed their potential to produce compounds with antimicrobial or anticancer activities. A specimen of L. ochroleuca was collected in a rocky shore in northern Portugal, and fragments of tissues from different parts of the macroalgae (holdfast, stipe, and blades) were surface sterilized and plated in three culture media selective for actinobacteria. A total of 90 actinobacterial strains were isolated, most of which affiliated with the genus Streptomyces. Isolates associated with the genera Isoptericola, Rhodococcus, Nonomuraeae, Nocardiopsis, Microbispora, and Microbacterium were also obtained. Organic extracts from the isolates were tested for their antimicrobial activity using the agar-based disk diffusion method, followed by determination of minimum inhibitory concentration (MIC) values. Forty-five isolates inhibited the growth of Candida albicans and/or Staphylococcus aureus, with MIC values ranging from

Classificada como uma categoria de Unidade de Conservação Brasileira de uso sustentável, a Reserva de Desenvolvimento Sustentável da Barra do Una (RDSBU) apresenta notória biodiversidade de plantas medicinais tradicionalmente utilizadas ao longo das gerações no tratamento de diversas doenças. Assim, o levantamento de dados etnofarmacológicos se faz essencial para documentar e preservar o conhecimento da população local desta área. O estudo teve como objetivo obter informações etnofarmacológicas sobre as plantas medicinais utilizadas pela população local da RDSBU. Os dados etnofarmacológicos foram coletados por meio de entrevista semiestruturada com 37 informantes da população da RDSBU. As espécies vegetais citadas foram coletadas por meio da técnica de visita guiada e foi realizada a identificação taxonômica. O fator de consenso do informante e o nível de fidelidade foram usados ​​para determinar a importância dos recursos de plantas medicinais. A validação farmacológica e uma revisão sistemática dos compostos fitoquímicos para as espécies mencionadas também foram realizadas. Para as espécies sem informação sobre os constituintes fitoquímicos foi realizada uma triagem fitoquímica. 103 plantas foram citadas, onde 70 puderam ser identificadas em nível de espécie. Entre as plantas que foram identificadas em nível de espécie, apenas 21 possuem validação farmacológica em farmacopéias e monografias. Tabebuia cassinoides (Lam.) DC. e Imperata tenuis Hack. não apresentaram informações na literatura sobre constituintes fitoquímicos e atividades biológicas e foram analisadas por meio de triagem fitoquímica. O presente estudo documentou pela primeira vez a etnofarmacologia de plantas medicinais da comunidade da RDSBU.

Publisher's version (útgefin grein), Obesity is a complex disease resulting in several metabolic co-morbidities and is increasing at epidemic rates. The marine environment is an interesting resource of novel compounds and in particular cyanobacteria are well known for their capacity to produce novel secondary metabolites. In this work, we explored the potential of cyanobacteria for the production of compounds with relevant activities towards metabolic diseases using a blend of target-based, phenotypic and zebrafish assays as whole small animal models. A total of 46 cyanobacterial strains were grown and biomass fractionated, yielding in total 263 fractions. Bioactivities related to metabolic function were tested in different in vitro and in vivo models. Studying adipogenic and thermogenic gene expression in brown adipocytes, lipid metabolism and glucose uptake in hepatocytes, as well as lipid metabolism in zebrafish larvae, we identified 66 (25%) active fractions. This together with metabolite profiling and the evaluation of toxicity allowed the identification of 18 (7%) fractions with promising bioactivity towards different aspects of metabolic disease. Among those, we identified several known compounds, such as eryloside T, leptosin F, pheophorbide A, phaeophytin A, chlorophyll A, present as minor peaks. Those compounds were previously not described to have bioactivities in metabolic regulation, and both known or unknown compounds could be responsible for such effects. In summary, we find that cyanobacteria hold a huge repertoire of molecules with specific bioactivities towards metabolic diseases, which needs to be explored in the future., This work was financed by national funds through FCT (Portugal), BMBF (Germany), Rannis (Iceland) and Formas (Sweden), within the framework of the European ERA-NET Marine Biotechnology project “CYANOBESITY—Cyanobacteria as a source of bioactive compounds with effects on obesity and obesity-related co-morbidities”. The research was additionally supported by the FCT strategic fund UID/Multi/04423/2019. Ralph Urbatzka and Tiago Ribeiro were supported by FCT grants SFRH/BPD/112287/2015 and SFRH/BD/139131/2018 respectively. SU received additional funding from the project Aging and Metabolic Programming (AMPro).

The cyclic peptides portoamides produced by the cyanobacterium Phormidium sp. LEGE 05292 were previously isolated and their ability to condition microcommunities by allelopathic effect was described. These interesting bioactive properties are, however, still underexplored as their biotechnological applications may be vast. This study aims to investigate the antifouling potential of portoamides, given that a challenge in the search for new environmentally friendly antifouling products is to find non-toxic natural alternatives with the ability to prevent colonization of different biofouling species, from bacteria to macroinvertebrates. A multi-bioassay approach was applied to assess portoamides antifouling properties, marine ecotoxicity and molecular mode of action. Results showed high effectiveness in the prevention of mussel larvae settlement (EC50 = 3.16 &micro, M), and also bioactivity towards growth and biofilm disruption of marine biofouling bacterial strains, while not showing toxicity towards both target and non-target species. Antifouling molecular targets in mussel larvae include energy metabolism modifications (failure in proton-transporting ATPases activity), structural alterations of the gills and protein and gene regulatory mechanisms. Overall, portoamides reveal a broad-spectrum bioactivity towards diverse biofouling species, including a non-toxic and reversible effect towards mussel larvae, showing potential to be incorporated as an active ingredient in antifouling coatings.

Crambe is an oilseed plant whose oil is suitable for various industrial purposes, such as production of biodiesel, plastics, rubbers, and cosmetics. There are reports of the rusticity of this species and its potential for growth in arid and semi-arid environments, where water scarcity and soil salinity are common. However, studies regarding the tolerance of its seeds to these stresses and the physiological responses triggered during germination and seedling establishment under these conditions are incipient. The objective of this study was to evaluate the tolerance of crambe seeds to water deficit and salt stress during germination and early seedling development, as well as to investigate the role of antioxidant enzymes on tolerance to these conditions. The seeds were exposed to water and salt stresses and their performance was analyzed. Seed water content, imbibition rate, percentage and speed of germination, and the base water potential for seed germination were evaluated. In the seedlings, the following measurements were made: root and shoot length, fresh and dry matter of roots and shoots, moisture content, and activity of the enzymes superoxide dismutase (SOD), catalase (CAT), and peroxidase (POX). Water deficit and salt stress reduce the water uptake rate of the seeds, the germination and growth of seedlings. The crambe seeds had higher tolerance to salt stress than to water deficit. Absorption of salt and the action of the SOD and CAT antioxidant enzymes are part of the tolerance mechanisms of crambe seeds to salt stress.

Early harvest may decrease seed exposure in the field after physiological maturation. The objective of this study was to evaluate the effect of glyphosate desiccant at different plant desiccation times at pre-harvest on the physiological quality of crambe (Crambe abyssinica Hochst) seeds. The experimental design was completely randomized in a 2 x 4 factorial scheme, involving the application of glyphosate desiccant in the plants at pre-harvest and the control (without plant desiccation) and four desiccation times, with three replicates per treatment. The desiccation of crambe plants at pre-harvest with glyphosate was efficient, positively affecting seed germination and vigor. Application of the desiccant in the plants, with 90% of brown seeds, at 103 days after sowing, allows the production of seeds with superior performance. RESUMO A antecipação da colheita pode diminuir a exposição das sementes no campo após a maturação fisiológica. Objetivou-se avaliar o efeito do dessecante glyphosate em diferentes épocas de dessecação das plantas em pré-colheita na qualidade fisiológica de sementes de crambe (Crambe abyssinica Hochst). Foi utilizado o delineamento experimental inteiramente casualizado em esquema fatorial 2 x 4, envolvendo a aplicação de dessecante glyphosate nas plantas em pré-colheita e a testemunha (sem dessecação das plantas), e quatro épocas de dessecação, com três repetições por tratamento. A dessecação das plantas de crambe em pré-colheita com glyphosate se mostrou eficiente, afetando positivamente a germinação e o vigor das sementes. A aplicação do dessecante nas plantas, com 90% das sementes de coloração marrom, aos 103 dias após a semeadura, possibilita a produção de sementes com desempenho superior.

International audience; Subtelomeres of most eukaryotes contain fast-evolving genes usually involved in adaptive processes. In common bean (Phaseolus vulgaris), the Co-2 anthracnose resistance (R) locus corresponds to a cluster of nucleotide-binding-site leucine-rich-repeat (NL) encoding sequences, the prevalent class of plant R genes. To study the recent evolution of this R gene cluster, we used a combination of sequence, genetic and cytogenetic comparative analyses between common bean genotypes from two distinct gene pools (Andean and Mesoamerican) that diverged 0.165 million years ago. Co-2 is a large subtelomeric cluster on chromosome 11 comprising from 32 (Mesoamerican) to 52 (Andean) NL sequences embedded within khipu satellite repeats. Since the recent split between Andean and Mesoamerican gene pools, the Co-2 cluster has experienced numerous gene-pool specific NL losses, leading to distinct NL repertoires. The high proportion of solo-LTR retrotransposons indicates that the Co-2 cluster is located in a hot spot of unequal intra-strand homologous recombination. Furthermore, we observe large segmental duplications involving both Non-Homologous End Joining and Homologous Recombination double-strand break repair pathways. Finally, the identification of a Mesoamerican-specific subtelomeric sequence reveals frequent interchromosomal recombinations between common bean subtelomeres. Altogether, our results highlight that common bean subtelomeres are hot spots of recombination and favor the rapid evolution of R genes. We propose that chromosome ends could act as R gene incubators in many plant genomes.

Barley-based diets contain a significant proportion of highly soluble 1,3-1,4-β-glucans that display anti-nutritive properties for monogastric animals, in particular for poultry. Cleavage of mixed linked glucans by the addition of exogenous enzymes leads to a significant reduction in the degree of polymerization of the polysaccharide and a consequent reduction of digesta viscosity. Plant cell wall 1,3-1,4-β-glucans may be hydrolyzed by 1,3-1,4-β-glucanases (EC 3.2.1.73) or by 1,4-β-glucanases (EC 3.2.1.4), generally termed cellulases, although it remains unknown which enzymes are more effective in the degradation of feed mixed linked anti-nutritive glucans. Here we compared the capacity of two recombinant Clostridium thermocellum enzymes, 1,3-1,4-β-glucanase 16A (CtGlc16A) and 1,4-β-glucanase 8A (CtCel8A), to improve the nutritive value of barley based diets for broilers. As already shown in a previous study ( Ribeiro et al., 2012 ), CtGlc16A is very effective to improve the performance of broilers fed with a highly viscous barley-based diet. In contrast, although remaining active and retaining its molecular integrity during passage through the GI tract, CtCel8A was unable to affect the nutritive value of the cereal based diet. In vitro studies revealed that CtGlc16A and CtCel8A are equally effective in reducing the viscosity of a purified 1,3-1,4-β-glucan solution. However, the capacity of CtCel8A to cleave 1,3-1,4-β-glucans is significantly affected by the presence of the barley-based feed. Taken together the results suggest that in vivo 1,4-β-glucanases tend to act preferentially on cellulosic substrates and not on mixed linked glucans. Although further work is required to extend these observations to other cellulases, the data suggest that exogenous 1,3-1,4-β-glucanases but not 1,4-β-glucanases are obligatory enzymes to improve the nutritive value of barley based diets for broilers.

Cnidarian toxic products, particularly peptide toxins, constitute a promising target for biomedicine research. Indeed, cnidarians are considered as the largest phylum of generally toxic animals. However, research on peptides and toxins of sea anemones is still limited. Moreover, most of the toxins from sea anemones have been discovered by classical purification approaches. Recently, high-throughput methodologies have been used for this purpose but in other Phyla. Hence, the present work was focused on the proteomic analyses of whole-body extract from the unexplored sea anemone Bunodactis verrucosa. The proteomic analyses applied were based on two methods: two-dimensional gel electrophoresis combined with MALDI-TOF/TOF and shotgun proteomic approach. In total, 413 proteins were identified, but only eight proteins were identified from gel-based analyses. Such proteins are mainly involved in basal metabolism and biosynthesis of antibiotics as the most relevant pathways. In addition, some putative toxins including metalloproteinases and neurotoxins were also identified. These findings reinforce the significance of the production of antimicrobial compounds and toxins by sea anemones, which play a significant role in defense and feeding. In general, the present study provides the first proteome map of the sea anemone B. verrucosa stablishing a reference for future studies in the discovery of new compounds. © 2018 by the authors. Acknowledgments: Dany Domínguez Pérez was supported by a Ph.D. grant (SFRH/BD/80592/2011) from the Portuguese Foundation for Science and Technology (FCT—Fundação para a Ciência e a Tecnologia, Portugal). A. Campos work were supported respectively by Postdoc grants SFRH/BPD/92978/2013 and SFRH/BPD/103683/2014 from the FCT. Armando A Rodríguez was supported by an Alexander von Humboldt postdoctoral fellowship (3.2-KUB/1153731 STP and the Collaborative Research Centre 1279 funded by the German Research Foundation). We are grateful to Isabel Cunha and Daniela Almeida from CIIMAR, FCUP, University of Porto, for the help in the identification, sampling and transporting of B. verrucosa specimens. To Barbara Frazão, from Instituto Português do Mar e da Atmosfera, Lisbon, Portugal for suggestions and by providing information related the species studied. This study was funded in part by the Strategic Funding UID/Multi/04423/2013 through national funds provided by FCT and the European Regional Development Fund (ERDF) in the framework of the program PT2020, by the European Structural and Investment Funds (ESIF) through the Competitiveness and Internationalization Operational Program—COMPETE 2020 and by National Funds through the FCT under the project PTDC/AAG-GLO/6887/2014 (POCI-01-0124-FEDER-016845), and by the Structured Programs of R&D&I INNOVMAR—Innovation and Sustainability in the Management and Exploitation of Marine Resources (NORTE-01-0145-FEDER-000035, Research Line NOVELMAR) and CORAL NORTE (NORTE-01-0145-FEDER-000036), and funded by the Northern Regional Operational Program (NORTE2020) through the ERDF.

Karyotype evolution in species with non-localised centromeres (holocentric chromosomes) is usually very dynamic and associated with recurrent fission and fusion (also termed agmatoploidy/symploidy) events. In Rhynchospora (Cyperaceae), one of the most species-rich sedge genera, all analysed species have holocentric chromosomes and their numbers range from 2n = 4 to 2n = 84. Agmatoploidy/symploidy and polyploidy were suggested as the main processes in the reshuffling of Rhynchospora karyotypes, although testing different scenarios of chromosome number evolution in a phylogenetic framework has not been attempted until now. Here, we used maximum likelihood and model-based analyses, in combination with genome size estimation and ribosomal DNA distribution, to understand chromosome evolution in Rhynchospora. Overall, chromosome number variation showed a significant phylogenetic signal and the majority of the lineages maintained a karyotype of 2n = 10 (~48% of the species), the most likely candidate for the ancestral number of the genus. Higher and lower chromosome numbers were restricted to specific clades, whilst polyploidy and/or fusion/fission events were present in specific branches. Variation in genome size and ribosomal DNA site number showed no correlation with ploidy level or chromosome number. Although different mechanisms of karyotype evolution (polyploidy, fusion and fission) seem to be acting in distinct lineages, the degree of chromosome variation and the main mechanisms involved are comparable to those found in some monocentric genera and lower than expected for a holocentric genus.

It is now well established that exogenous β-1,4-xylanases improve the nutritive value of wheat-based diets for poultry. Among other factors, the mechanism of action of exogenous enzymes may involve a microbial route resulting from the generation of prebiotic xylo-oligosaccharides (XOS) in the birds’ gastro-intestinal (GI) tract. In a series of three experiments, the effect of XOS on the performance of broilers fed wheat or corn-based diets was investigated. In experiment 1, birds receiving diets supplemented with XOS displayed an increased weight gain (P = 0.08). The capacity of XOS to improve the performance of animals during a longer trial (42 d) was investigated (Experiment 2). The data revealed that diet supplementation with XOS, tested at two incorporation rates (0.1 and 1 g/kg), or with an exogenous β-1,4-xylanase resulted in an increased nutritive value of the wheat-based diet. An improvement in animal performance was accompanied by a shift in the microbial populations colonizing the upper portions of the GI tract. XOS were also able to improve the performance of broilers fed a corn-based diet, although the effects were not apparent at incorporation rates of 10 g/kg. Together these studies suggest that in some cases the capacity of β-1,4-xylanases to improve the nutritive value of wheat-based diets is more related to their ability to produce prebiotic XOS than to their ability to degrade arabinoxylans. The extremely low quantities of XOS used in this study also challenge the depiction of a prebiotic being a quantitatively fermented substrate. These data also bring into question the validity of the “cell wall” mechanism, as XOS elicited an effect with clearly no action on endosperm cell wall integrity and yet the performance effects noted were equivalent or superior to the added enzymes.

The attack behavior of locust swarm has inspired the recently developed computer algorithmic model of locust swarm optimizer (LS) within the swarm artificial intelligence methods. Some NP-hard problems of combinatorial optimization that LS may solve are the following: the traveling salesman problem, the knapsack problem, routing in telecommunication networks, and cellular robotic systems. General models of these combinatorial optimization problems which can be solved with locust swarm optimizer technique are proposed in the present work.

The objective of this research is to study the maturation process of buffel grass cv. Aridus seeds in order to determine their best harvest season, aiming at maximum seed quality. During the inflorescence emergence phase, they were labeled upon showing from five to ten inflorescences emerged per m2. The first seed harvest was done 20 days after inflorescence emergence, and the other harvests were done at 5-day intervals, totaling six harvests. In each harvest season, the seeds were assessed as to water content, dry matter mass, germination, first germination count, seedlings emergence, emergence speed index, and electrical conductivity. The physiological maturity of buffel grass seeds (maximum germination, vigor and dry matter accumulation) occurs from 38 to 42 days after inflorescence emergence, moment when harvest must be done.

It is now well established that exogenous microbial β-glucanases effectively contribute to improve the nutritive value of barley based diets for broilers while they are predominantly effective in the first stages of the production cycle. Here, the performance of broilers fed on barley-based diets supplemented with a β-glucanase enzyme mixture, in part or during the entire length of the production cycle, was evaluated. Six hundred 1-day-old Ross 308 chicks were randomly divided into 4 groups. Animals were fed on a barley-based diet supplemented with a commercial enzyme cocktail for the entire duration of the trial (35 days) or exclusively during the first 11 or 23 days of the experiment. Animals of the control group were fed on the basal diet not containing the microbial biocatalyst mixture. Body weight and feed intake were recorded weekly and weight gain and feed conversion ratios were determined. At the end of the trial, 2 animals were slaughtered to collect gastrointestinal contents to evaluate viscosity and enzymatic activity. Weight/length of gastrointestinal compartments were also recorded. Data revealed that all β-glucanase supplemented groups outperformed non-supplemented birds at day 35. Significant differences in body weight were detected at day 11, suggesting that response to enzyme supplementation occurs, primarily, during the initial stages of broiler growth. In addition, final body weight of animals fed on diets supplemented during the first 11 or 23 days of the experiment was identical to animals supplemented during the entire trial (P>0.05). Together the data suggest that in barley-based diets exogenous enzymes act primarily in the earliest periods of broilers growing period and that enzyme supplementation may be restricted to the first 11 days of the production cycle without negatively affecting animal performance. Zymogram analysis allowed detecting β-glucanase activity in the crop of non-supplemented birds at day 35. This activity does not arise from endogenous enzymes present in the feed suggesting colonization of the crop with a β-glucanase secreting microbial population. Although this activity was not sufficient to reduce digesta viscosity in the upper portions of the GI tract when compared with birds exposed during the entire period of the experiment to the exogenous enzymes, it might have contributed to attenuate the antinutritive effects of β-glucans at later stages of animal growth.

O objetivo do trabalho foi caracterizar a morfologia de frutos, sementes e plântulas, assim como determinar o substrato adequado para germinação e desenvolvimento inicial de plântulas de vinagreira. No estudo morfológico de frutos, foram observadas características internas e externas referentes à coloração, consistência, forma, número de sementes e deiscência. Para descrição das sementes, analisou-se a coloração, forma, textura e consistência dos tegumentos. Foram conduzidos testes de germinação nos substratos rolo de papel “germitest”, entre e sobre papel mata-borrão e entre areia, visando à caracterização das plântulas e a avaliação da qualidade fisiológica das sementes, sendo avaliadas quanto à germinação e vigor (índice de velocidade de germinação, comprimento de plântulas, massa fresca e seca de plântulas). O estudo morfológico das estruturas de vinagreira é importante para o conhecimento da espécie, favorecendo o desenvolvimento de técnicas adequadas de manejo. A germinação é epígea fanerocotiledonar, iniciando a partir do segundo dia após semeadura. Recomenda-se utilizar o substrato rolo de papel germistest para o teste de germinação em sementes de vinagreira com contagens aos sete (primeira contagem) e 14 dias (germinação) após a semeadura.

Cabo Verde is located on the West African coast, was discovered uninhabited by Portuguese explorers, colonized by European and African slaves and is culturally characterized by a mixture of their settlers. Cabo Verde is one of the countries with the highest number of emigrants living in Portugal. In order to validate the genetic markers included in the Combined DNA Index System and in the European Standard Set, samples from 500 unrelated individuals from Cabo Verde, currently living in Lisboa, were studied with the PowerPlex ® Fusion System. To implement the genotyping method at the Servico de Genetica e Biologia Forenses (SGBF) we determined the minimum final volume of the PCR reaction, the proper number of PCR cycles and the minimum DNA concentration to obtain a complete genetic profile. Stutter identification and its proportion and allelic imbalance within markers were also determined. Results were compared with genetic profiles obtained by the current PCR kits used in SGBF routine testing verifying concordance between common loci. There were no off-ladder alleles detected and it was possible to type microvariants which were not present in the allelic ladders of other PCR kits. PowerPlex ® Fusion System allows for the simultaneous characterization of Combined DNA Index System and in the European Standard Set genetic markers, in an economically way, with greater sensitivity and solid results.

SUMMARY Triticale, a hybrid of wheat and rye, is a cereal grain that displays a nutritive value similar to wheat and a resistance to hard agronomic conditions, which are more identical to rye. Because of its lower price in comparison with wheat or corn, triticale may be a potential alternative raw material for broiler diets. However, triticale is known to contain a significant proportion of soluble arabinoxylans, which are known to act as antinutritive factors in poultry. Nevertheless, levels of soluble polysaccharides in triticale are more similar to wheat and are not as high as in rye. In the present study, the capacity of xylanases, used at different levels, to improve the nutritive value of a triticale-based diet for broilers was investigated. On the basis of these data, triticale may be used effectively as the main feedstuff in broiler diets. However, supplementation with xylanase improved the nutritive value of the cereal-based diet when incorporated at 75% (wt/wt) of the recommended dose. Xylanases may modulate bird performance through the microflora route. As expected, incorporation of the exogenous enzymes reduced digesta viscosity. However, xylanase incorporation at 100 and 200% of the recommended levels did not have an effect on bird performance.

In this paper, we present a study of consumer preferences for mobile telecommunications plans and operator characteristics. The objective of the study was to identify consumer preferences for the following characteristics of mobile plans: the importance of using the same provider as friends and family (calling club network effects), the market share of the provider (pure network effects), the length of the commitment period, monthly fee/recharge obligations and per minute call charges for calls made within and outside the provider network. A discrete choice experiment was used as a preference elicitation method and implemented in face-to-face interviews. We present results regarding willingness to pay for the described features as well as their relationship to socio-demographic variables. Consumers are willing to pay 1.3 Euro per month more to reduce the commitment period from 1 year to 6 months and willing to pay 2.5 euros per month more to be part of a larger network. Consumers are also twice as much more sensitive to within-network price variations than extra-network price variations. These results remain unchanged in the sub-sample of those that have internet access suggesting that a web-based surveys are capable of producing unbiased results. The implications of these results for regulatory policy are discussed. Discrete choice experiment study eliciting mobile telecom plans and operator preferences.Features included: network effects, commitment period, monthly fee and on-net/off-net charges.Consumers are willing to pay 1.3 per month more to reduce by 6 months the commitment period.Consumers are willing to pay 2.5 per month more to be part of a larger network.Those consumers who are willing to pay are twice as much more sensitive to on-net price than to off-net price.Regulatory policy implications are discussed.

Common bean (Phaseolus vulgaris) subtelomeres are highly enriched for khipu, the main satellite DNA identified so far in this genome. Here, we comparatively investigate khipu genomic organization in Phaseolus species from different clades. Additionally, we identified and characterized another satellite repeat, named jumper, associated to khipu. A mixture of P. vulgaris khipu clones hybridized in situ confirmed the presence of khipu-like sequences on subterminal chromosome regions in all Phaseolus species, with differences in the number and intensity of signals between species and when species-specific clones were used. Khipu is present as multimers of similar to 500 bp and sequence analyses of cloned fragments revealed close relationship among khipu repeats. The new repeat, named jumper, is a 170-bp satellite sequence present in all Phaseolus species and inserted into the nontranscribed spacer (NTS) of the 5S rDNA in the P. vulgaris genome. Nevertheless, jumper was found as a high-copy repeat at subtelomeres and/or pericentromeres in the Phaseolus microcarpus lineage only. Our data argue for khipu as an important subtelomeric satellite DNA in the genus and for a complex satellite repeat composition of P. microcarpus subtelomeres, which also contain jumper. Furthermore, the differential amplification of these repeats in subtelomeres or pericentromeres reinforces the presence of a dynamic satellite DNA library in Phaseolus.

The mountain bike is one of the most used equipment's in outdoor sports activities. The present work presents a distributed sensing system expressed by a WSN for cycling assessment to increase performance of athletes involved in this probes. The implemented system performs acquisition, processing and data communication based on ZigBee protocol that also provides the machine-to-machine communication and data storage in a cloud platform. The information stored on the cloud server can be accessed through a mobile application that analyses and correlates all metrics calculated using the training data obtained during training session. The system permits to record the athletes training data and to access it in order to improve their performances during the next competitions based on data analysis. The recorded data of the evolution of each athlete permits to perform appropriate comparisons between different training sessions and different athletes results.

Satellite DNA repeats (or satDNA) are fast-evolving sequences usually associated with condensed heterochromatin. To test whether the chromosomal organisation of centromeric and non-centromeric satDNA differs in species with holocentric chromosomes, we identified and characterised the major satDNA families in the holocentric Cyperaceae species Rhynchospora ciliata (2n = 10), R. globosa (2n = 50) and R. tenuis (2n = 2x = 4 and 2n = 4x = 8). While conserved centromeric repeats (present in R. ciliata and R. tenuis) revealed linear signals at both chromatids, non-centromeric, species-specific satDNAs formed distinct clusters along the chromosomes. Colocalisation of both repeat types resulted in a ladder-like hybridisation pattern at mitotic chromosomes. In interphase, the centromeric satDNA was dispersed while non-centromeric satDNA clustered and partly colocalised to chromocentres. Despite the banding-like hybridisation patterns of the clustered satDNA, the identification of chromosome pairs was impaired due to the irregular hybridisation patterns of the homologues in R. tenuis and R. ciliata. These differences are probably caused by restricted or impaired meiotic recombination as reported for R. tenuis, or alternatively by complex chromosome rearrangements or unequal condensation of homologous metaphase chromosomes. Thus, holocentricity influences the chromosomal organisation leading to differences in the distribution patterns and condensation dynamics of centromeric and non-centromeric satDNA.

AimThe contribution of insulin resistance (IR) and glucose tolerance to the maintenance of blood glucose levels in non diabetic pregnant Wistar rats (PWR) was investigated.Main methodsPWR were submitted to conventional insulin tolerance test (ITT) and glucose tolerance test (GTT) using blood sample collected 0, 10 and 60min after intraperitoneal insulin (1U/kg) or oral (gavage) glucose (1g/kg) administration. Moreover, ITT, GTT and the kinetics of glucose concentration changes in the fed and fasted states were evaluated with a real-time continuous glucose monitoring system (RT-CGMS) technique. Furthermore, the contribution of the liver glucose production was investigated.Key findingsConventional ITT and GTT at 0, 7, 14 and 20days of pregnancy revealed increased IR and glucose tolerance after 20days of pregnancy. Thus, this period of pregnancy was used to investigate the kinetics of glucose changes with the RT-CGMS technique. PWR (day 20) exhibited a lower (p

To improve the nutritive value of barley-based diet for broilers, 2 experiments using 2 different barley lots were performed to evaluate the capacity of a mesophilic cellulase when fused to a β-glucan specific family 11 carbohydrate-binding module. The data revealed that the recombinant β-glucanase derivatives were not appropriate for feed supplementation because of a lack of stability at acidic pH levels. However, under the same experimental conditions, a commercial enzyme mixture improved the nutritive value of 1 of the cereal lots used. Analysis of the nutritive value of the 2 barleys revealed intrinsic differences in the levels of endogenous β-glucanase activity. These differences were extensively evident when the studies were expanded to a range of 64 barley lots. Thus, to clarify the effect of endogenous cellulases on the efficacy of exogenous β-glucanases used to supplement barley-based diets for poultry, 2 barley lots presenting low and high levels of endogenous plant cell wall-degrading enzymes were selected. These lots were used to prepare 2 barley-based diets, which were supplemented with or without a commercial enzyme product and fed to broiler chicks. The data revealed that the exogenous enzymes were effective when the basal diet presented low levels of endogenous β-glucanases but were unable to improve the nutritive value of the barley lot displaying higher β-glucanase activity. Thus, these studies suggest that levels of endogenous β-glucanases may affect the efficacy of exogenous enzymes used to improve the nutritive value of barley-based diets for broilers. The development of a quick β-glucanase assay that could be applied for cereal-based feeds may help identify those barley-based diets that are more responsive to the action of feed enzymes.

A cytogenetic map of common bean was built by in situ hybridization of 35 bacterial artificial chromosomes (BACs) selected with markers mapping to eight linkage groups, plus two plasmids for 5S and 45S ribosomal DNA and one bacteriophage. Together with three previously mapped chromosomes (chromosomes 3, 4, and 7), 43 anchoring points between the genetic map and the cytogenetic map of the species are now available. Furthermore, a subset of four BAC clones was proposed to identify the 11 chromosome pairs of the standard cultivar BAT93. Three of these BACs labelled more than a single chromosome pair, indicating the presence of repetitive DNA in their inserts. A repetitive distribution pattern was observed for most of the BACs; for 38% of them, highly repetitive pericentromeric or subtelomeric signals were observed. These distribution patterns corresponded to pericentromeric and subtelomeric heterochromatin blocks observed with other staining methods. Altogether, the results indicate that around half of the common bean genome is heterochromatic and that genes and repetitive sequences are intermingled in the euchromatin and heterochromatin of the species. Electronic supplementary material The online version of this article (doi:10.1007/s10577-010-9129-8) contains supplementary material, which is available to authorized users.

1. Exogenous microbial beta-1,3-1,4-glucanases and hemicellulases contribute to improving the nutritive value of cereals rich in soluble non-starch polysaccharides for poultry. 2. In general, plant cell wall hydrolases display a modular structure comprising a catalytic module linked to one or more non-catalytic carbohydrate-binding modules (CBMs). Based on primary structure similarity, CBMs have been classified in 50 different families. CBMs anchor cellulases and hemicellulases into their target substrates, therefore eliciting efficient hydrolysis of recalcitrant polysaccharides. 3. A study was undertaken to investigate the effects of a family 11 beta-glucan-binding domain in the function of recombinant derivatives of cellulase CtLic26A-Cel5E of Clostridium thermocellum that were used to supplement a barley-based diet at lower dosage rates. 4. The results showed that birds fed on diets supplemented with the recombinant CtLic26A-Cel5E modular derivative containing the family 11 CBM or the commercial enzyme mixture Rovabio Excel AP tended to display improved performance when compared to birds fed diets not supplemented with exogenous enzymes. 5. It is suggested that at lower than previously reported enzyme dosage (10 U/kg vs 30 U/kg of basal diet), the beta-glucan-binding domain also elicits the function of the recombinant CtLic26A-Cel5E derivatives. 6. Finally, the data suggest that exogenous enzymes added to barley-based diets act primarily in the proximal section of the gastrointestinal tract.

1. Cellulases and xylanases display a modular architecture that comprises a catalytic module linked to one or more non-catalytic carbohydrate-binding modules (CBMs). CBMs have been classified into 52 different families, based on primary structure similarity. These non-catalytic modules mediate a prolonged and intimate contact of the enzyme with the target substrate eliciting efficient hydrolysis of the target polysaccharides. 2. A study was undertaken to investigate the importance of a family 11 CBM, displaying high affinities for barley beta-glucans, in the function of recombinant derivatives of cellulase CtLic26A-Cel5E of Clostridium thermocellum used to supplement a barley-based diet for broiler chicken. 3. The results showed that birds fed on diets containing the recombinant CtLic26A-Cel5E modular derivatives or the commercial enzyme mixture Rovabio Excel AP displayed improved performance when compared with birds fed on diets not supplemented with exogenous enzymes. 4. It is suggested that the enzyme dosage used in this study (30 U/kg of basal diet), was probably too high for the efficacy of the family 11 CBM to be noticed. It remains to be established if the targeting effect resulting from the incorporation of CBMs in plant cell wall hydrolases may be effective at lower exogenous enzyme dosages.

The season of harvest is one of the most important factors influencing seed quality. This study evaluated the effect of different harvesting times on the physiological quality of Coriandrum sativum L. seeds, cv. Palmeira. The experimental design was completely randomized, with six treatments (harvest times) and four replications. The experiment was conducted during the months of April to October 2010. The first moment to harvest seeds was carried out 15 days after full bloom, when approximately 50% of plants were in flowering phase. Other harvests (22, 29, 36, 43 and, 50 days after bloom) were carried out at 7 days intervals until the seeds reached 16.0% of moisture, which occurred in the 6th harvest. Immediately after each harvest, the seeds were analysed for water content, germination and vigor (first count, seedling emergence and emergence rate index). The harvest season most suitable for C. sativum seeds cv. Palmeira occurred between 46 and 50 days after flowering, when the seeds have the highest physiological quality and moisture content between 20 and 16%. RESUMO A época de colheita é um dos fatores mais importantes que influencia a qualidade das sementes. Este estudo avaliou o efeito de diferentes épocas de colheita na qualidade fisiológica de sementes de Coriandrum sativum L., cv. Palmeira. O delineamento experimental foi inteiramente casualizado, com seis tratamentos (épocas de colheita) e quatro repetições. O experimento foi conduzido durante os meses de abril a outubro de 2010, em que a primeira época de colheita das sementes foi realizada aos 15 dias após a floração quando aproximadamente 50% das plantas apresentavam flores. As demais colheitas (22, 29, 36, 43 e 50 dias após floração) foram realizadas em intervalos de 7 dias até as sementes atingirem aproximadamente 16% de umidade, o que ocorreu na 6ª colheita. Imediatamente após cada colheita as sementes foram avaliadas quanto ao teor de água, germinação e vigor (primeira contagem, emergência de plântulas e índice de velocidade de emergência). A época mais adequada para a colheita de sementes de C. sativum cv. Palmeira ocorreu entre 46 e 50 dias após a floração quando as sementes apresentaram a mais alta qualidade fisiológica e teor de umidade entre 20 e 16%.

The relative importance of explant, cytokinin type, carbon source and gelling agent for annatto organogenesis was studied. The best organogenic response, including adventitious shoot number and elongation, was obtained when hypocotyl segments and rooted hypocotyls were cultured onto MS medium supplemented with 4.56 μM zeatin, 87.6 mM sucrose, and 2.8 g l−1 Phytagel®. Adventitious shoots derived from hypocotyl segments were less frequent and more difficult to elongate than those derived from rooted hypocotyls. Thidiazuron (TDZ) promoted a higher organogenic response in rooted hypocotyls, resembling a rosette-like structure, but impaired shoot elongation. Histological investigation showed that zeatin-induced meristemoids originated mainly from wounding tissues, and that TDZ induced a high level of mitotic division resulting in several proliferation zones nearby the epidermis and outer cortical tissues. Rhizogenesis efficiency (rooting frequency and root number) was greater at the highest indole-3-butyric acid (IBA) concentration (5.0 μM) employed, although calli occurred at the basal end of shoots. Eighty percent of rooted plantlets survived after acclimatization. This optimized regeneration protocol may enable further development of an efficient genetic transformation protocol for this species.

1. Chicken breast meat is a lean meat due to its low content of intramuscular fat (IMF) resulting in an overall lower acceptability by consumers due to a decrease in juiciness, flavour and increased chewiness. Recently, studies performed in pigs suggested the possibility of increasing IMF by decreasing dietary crude protein (CP) content, an effect possibly mediated through an increased lipogenesis. 2. Dietary supplementation with lipids rich in omega 3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA) may modulate an increase in the content of these fatty acids in meat from monogastric animals and, thus, promote the daily intake of n-3 LC-PUFA by humans. 3. LC-PUFA are very susceptible to oxidation, resulting in off-flavours that affect meat quality and consumers' acceptability. 4. This trial was conducted to assess the effect of reducing dietary CP, from 21% to 17%, on chicken's meat IMF content and, simultaneously, to evaluate if a complementary supplementation with a proprietary n-3 LC-PUFA source (DHA Gold™) could improve meat quality. These effects were assessed by measuring productive performance and meat quality, oxidative stability, sensory traits and fatty acid profile. 5. A reduction in CP content of broiler diets, from 21% to 17%, balanced for lysine, improved performance while it was not sufficient to increase IMF content in chicken meat. In contrast, DHA Gold™ supplementation had a positive impact both in broiler productive parameters and in meat fatty acid profile. 6. In addition, incorporation of 7.4% of DHA Gold™ in the diet promoted carcass yield but negatively affected chicken meat acceptability by consumers, due to a decrease of meat oxidative stability. 7. Overall the data suggest that neither a dietary supplementation with DHA Gold™ nor a reduction in CP have a direct positive effect in the levels of IMF present in broiler meat.

For adapting well to various soil and climatic conditions in Brazil and the tradition of high consumption, bean plays a key role in feeding the population. Despite the different environmental conditions, there are few specific cultivars for each planting season, particularly when aimed at the production of quality seeds. The objective of this study was to evaluate the physiological potential of bean seeds from five population densities, under the edafoclimatic conditions of Janauba, northern Minas Gerais. The experiment was conducted on winter crop season, with planting in August 2011. The experimental design was a randomized block in 3 x 5 factorial, with four replicates. The treatments consisted of three cultivars of common bean with different growth types (Ouro Vermelho, Ouro Negro and BRSMG Madreperola, and five densities (100, 200, 300, 400 and 500 thousand plants ha -1 ). Seeds were evaluated for water content, germination and vigor (tests of first count, seedling emergence, emergence speed and accelerated aging). From the results, it can be concluded that the bean seeds of the cultivar Ouro Negro present superior physiological potential in relation to the cultivars Ouro Vermelho and BRSMG Madreperola. Increasing the sowing density up to 300 thousand plants ha -1 is suitable for the production of bean seeds on winter crop season, representing a good alternative for cultivation in the region.

1. Microbial β-1,3-1,4-glucanases improve the nutritive value of barley-based diets for poultry by effectively decreasing the degree of polymerisation of the anti-nutritive soluble β-glucans. Glycoside hydrolases (GHs) acting on recalcitrant polysaccharides display a modular architecture comprising a catalytic domain linked to one or more non-catalytic Carbohydrate-Binding Modules (CBMs). 2. GHs and CBMs have been classified in different families based on primary structure similarity (see CAZy webpage at http://www.cazy.org ). The role of CBMs is to anchor the appended GHs into their target substrates, therefore eliciting the efficient hydrolysis of structural carbohydrates. 3. Here we describe the biochemical properties of the family 16 GH from Clostridium thermocellum, termed CtGlc16A. CtGlc16A is a thermostable enzyme that specifically acts on β-1,3-1,4-glucans with a remarkable catalytic activity (38000 U/mg protein). 4. CtGlc16A, individually or fused to the family 11 β-glucan-binding domain of cellulase CtLic26A-Cel5E of C. thermocellum, was used to supplement a highly viscous barley-based diet for broilers. 5. The data showed that birds fed on diets supplemented with the recombinant enzymes displayed an improved performance when compared with birds given diets not supplemented with exogenous enzymes. However, inclusion of the non-catalytic CBMs had no influence on the capacity of CtGlc16A to reduce the anti-nutritive effects of soluble β-1,3-1,4-glucans. 6. The data suggest that at elevated dosage rates, CBMs might be unable to potentiate the catalytic activity of appended catalytic domains; this effect may only be revealed when feed enzymes are incorporated at low levels.

In this work the genetic divergence among 14 sweet cassava cultivars was estimated by their morphological agronomic traits and RAPD molecular markers. The Tocher cluster analysis and the Nearest Neighbor Method were applied. The most dissimilar cultivars were Pão and Guaíra, Fécula Branca and Pão, and Pão and Caipira, while the most similar cultivar were the Fécula Branca and Branca 1, Branca 3 and Branca 1, and Guaíra and Branca 1. The Jaccard's coefficient showed that the most similar cultivars were Guaíra and Quarenta Quilos, while the most dissimilar were Branca 3 and Amarela da Rama Cinza. The divergence analysis indicated that promising crosses could be made between the Branca 3 cultivar and the Pão, Amarela 1, Fécula Branca and Amarela 2 cultivars for the high genetic divergence, favorable agronomic and culinary traits, and disease resistance on the part of at least one of the parents involved in the cross.A divergência genética entre 14 cultivares de mandioca-de-mesa foi estimada mediante o uso de caracteres morfoagronômicos e de marcadores moleculares RAPD. As análises de agrupamento de Tocher e do Vizinho Mais Próximo foram efetuadas. As cultivares mais divergentes foram Pão e Guaíra, Fécula Branca e Pão, e Pão e Caipira, enquanto as mais similares foram Fécula Branca e Branca 1, Branca 3 e Branca 1, e Guaíra e Branca 1. O coeficiente de Jaccard indicou que as cultivares mais similares foram Guaíra e Quarenta Quilos, enquanto as mais dissimilares foram Branca 3 e Amarela da Rama Cinza. A análise de divergência indicou que cruzamentos promissores deveriam ser efetuados entre as cultivares Branca 3, Pão, Amarela 1, Fécula Branca e Amarela 2 pela alta divergência genética, características agronômicas e culinárias favoráveis, e resistência à doenças de parte de pelo menos um dos parentais envolvidos no cruzamento.

O conhecimento das condições adequadas de água e temperatura é de fundamental importância, para as atividades metabólicas envolvidas no processo germinativo. Entretanto, as Regras para Análise de Sementes não indicam o volume de água e a temperatura ideais para a germinação de sementes de pitaya. O trabalho foi conduzido com o objetivo de avaliar diferentes volumes de água no substrato e temperaturas na germinação e no vigor de sementes de pitaya. O delineamento experimental utilizado foi inteiramente casualizado em esquema fatorial 5 x 3, com os tratamentos constituídos pela combinação de cinco volumes de água no substrato e três temperaturas. Os volumes de água utilizados foram equivalentes a 1,5; 2,0; 2,5; 3,0 e 3,5 vezes o peso do substrato seco. Foram testadas as temperaturas constantes de 20; 25 e 30 °C. Avaliou-se a germinação, primeira contagem, índice de velocidade de germinação, comprimento e massa seca de plântulas. A temperatura de 25 °C e o volume de água na faixa de 2,5 e 3,5 vezes o peso do substrato é a combinação mais adequada para a condução dos testes de germinação e vigor das sementes de pitaya. A temperatura de 20 °C e o volume de água de 1,5 vezes o peso do substrato inibiu a germinação das sementes. Knowledge of appropriate conditions of water and temperature is very important for the metabolic activities involved in the germination process. However, the Rules for Testing Seeds do not indicate the volume of water and the ideal temperature for pitaya germination. The study was carried out to evaluate different water volumes in the substrate and temperatures on seeds germination and vigor of pitaya. The experimental design was completely randomized in a factorial 5 x 3, with treatments consisted of five water volumes in the substrate and three temperatures. The used water volumes was equivalent to 1.5; 2.0; 2.5; 3.0 and 3.5 times the dry weight of the substrate. Were tested at constant temperatures of 20, 25 and 30 °C. The parameters evaluated were: germination, first count of germination, emergence speed index, length and dry mass of plantlets. The temperature of 25 °C and a water volume in the range of 2.5 to 3.5 times the weight of substrate is the most suitable combination for doing germination tests and evaluating the vigor of pitaya seeds. The temperature of 20 °C and the volume of water 1.5 times the weight of the substrate inhibited germination of seeds.

In Portugal, the telecommunications incumbent offers broadband access to the Internet, both through digital subscriber line and cable modem. In this article, we estimate the impact on broadband access to the Internet of the structural separation of these two businesses. We use a panel of consumer level data and a discrete choice model to estimate the price elasticities of demand and the marginal costs of broadband access to the Internet. Based on these estimates, we simulate the effect on prices and social welfare of the structural separation. Our results indicate that the structural separation would lead to substantial price reductions. For broadband clients, on average, each household would save 3:37 euros per month, or 14% of the current price levels. Overall, on average, each household would save 2:73 euros per month, or 14% of the current price levels. We test the robustness of our results in terms of: (i) the estimates of the demand elasticities, (ii) the strategic behavior of the firms, and (iii) the market share estimates. There is no evidence of collusion.

Embora existam outros aspectos relacionados, como clima, solo e manejo da cultura, o hábito de crescimento da cultivar é um dos principais fatores envolvidos na resposta do feijoeiro à densidade de semeadura. Para tanto, o emprego de densidade de semeadura adequada é fator fundamental à obtenção de melhores respostas tecnológicas que resultem em maiores produtividades de grãos na cultura do feijão. Com o objetivo de estudar os efeitos de densidades de semeadura e safras de cultivo sobre o rendimento de grãos e componentes primários de rendimento de cultivares de feijoeiro, foi conduzido um experimento nas safras da seca e de inverno. Os tratamentos foram dispostos em esquema fatorial 4x5, envolvendo quatro cultivares de feijoeiro (Ouro Vermelho, Ouro Negro, Madrepérola e Manteigão Vermelho) e cinco densidades de semeadura (100, 200, 300, 400 e 500 mil plantas ha^ -1 ). Em ambas as safras, o delineamento experimental utilizado foi o de blocos casualizados, com quatro repetições. Por ocasião da colheita foi avaliado o estande final, o rendimento de grãos e os componentes primários (número de vagens por planta, número de grãos por vagem e massa de cem grãos). Realizou-se análise de variância conjunta para safras, e os efeitos de densidade de semeadura foram submetidos à análise de regressão, já os efeitos de cultivares foram estudados pelo teste de Tukey e os de safras pelo teste F, a 1 e a 5%. Com exceção do cultivar Ouro Negro cultivado na safra de inverno, em que o maior rendimento de grãos é obtido com população em torno de 300 mil plantas por hectare, o aumento na densidade de semeadura não interfere na produtividade do feijoeiro. Independentemente da densidade de semeadura, o feijoeiro cultivado na safra da seca apresenta maior número de grãos por vagem, maior massa média de cem grãos do que o cultivado na safra de inverno. While there are others aspects associated, such as climate, soil and crop management, the growth habit of the cultivar is one of the main factors involved in answer in gof common bean to sowing density. Thus, the use of appropriate seeding rate is critical to obtaining the best technological solutions that result inhigher grain yields in the common bean. With goal of estudy the effects of seeding density and seasons about yield of grains and primary components of yield of common bean cultivars, was conduced a experiment in the experimental farm of the UNIMONTES, in Janaúba, MG, in the dry seanson and winter season in the year of 2011. The treatments were arranged in factorial scheme 4x5, involving four cultivars of bean (Ouro Vermelho, Ouro Negro, Madrepérola e Manteigão Vermelho) and five seeding density (100, 200, 300, 400 e 500 thousand plants ha^-1). In both seasons, the experimental design utilized was randomized complete block, with four replications. At harvest was evaluated the final stand, yield of grains e primary components (number of pods by plant, number of grain by pod and mass of hundred grains). It was realized joint analysis of data for seasons of growth, and the effects of sowing density were subjected to regression analysis, however the effects of cultivars were studied by Tukey test and seasons by F test at 1and 5%. With the exception of Ouro Negro, grown in the winter season, which the highest grain yield is obtained with sowing density around 300 thousand plants by hectare, the increase in density of seeding does not affect the yield of common bean under the conditions considered in this study. Irrespective of sowing density, the common bean cultivated in the dry season show a greater number of seeds by pod, greater average weight of hundred grains, than common bean grown in the winter season.