Vicilinas de sementes de leguminosas selvagens:purificação, caracterização, efeito de deletério e mecanismo de ação para bruquídeos e para fungos fitopatogênicos

Globulins fractions of legume seeds of Crotalaria pallida, Erytrina veluntina and Enterolobium contortisiliquum were isolated and submitted to assays against serine, cysteine and aspartic proteinases, as also amylase present in midgut of C. maculatus and Z. subfasciatus. Hemagglutination assays indicated presence of a lectin in E. veluntina globulin fractions. This lectin had affinity to human erythrocytes type A, B and O. Vicilins were purified by chromatography on Sephacryl S-300 followed of a chromatography on Sephacryl S-200, which was calibrated using protein markers. Vicilins from C. pallida (CpV) and E. veluntina (EvV) seeds had a molecular mass of 124.6 kDa and E. contortisiliquum a molecular mass of 151kDa. Eletrophoresis in presence of SDS showed that CpV was constituted by four subunities with apparent molecular mass of 66, 63, 57 and 45 kDa, EvV with three subunities with apparent molecular mass of 45kDa and EcV four subunities, two with 37.1 kDa and two with 25.8 kDa. Non denaturantig eletrophoresis displayed single bands with high homogeneity, where CpV had lower acidic behavior. All vicilins are glycoproteins with carbohydrate contents at 1 to1.5%. Bioassays were done to detect deleterious effects of vicilins against C. maculatus and Z. subfasciatus larvae. CpV, EvV and EcV exhibited a WD50 of 0.28, 0.19 and 1.03%; LD50 0.2, 0.26, and 1.11% respectively to C. maculatus. The dose responses of CpV, EvV and EcV to Z. subfasciatus were: WD50 of 0.12, 0.14, 0.65% and LD50 of 0.09, 0.1, and 0.43% respectively. The mechanism of action of these proteins to bruchids should be based on their properties of bind to chitin present in mid gut of larvae associated with the low digestibility of vicilin. In assays against phytopatogenous fungus, only EcV was capable of inhibit F. solani growth at concentrations of 10 and 20 µg and its action mechanism should be also based in the affinity of EcV to chitin present in the fungi wall

Vicilina de sementes da leguminosa selvagem Anadenanthera macrocarpa: purificação, caracterização, efeito deletério e mecanismo de ação para Callosobruchus maculatus

Grains and legume seeds are foods that form the basis of the diets of many cultures around the world, winch contritbute to the daily nutrient requirements of humans. Vicilins (7S globulin) are storage proteins found in legume seeds, and may have an additional function constitutive defense of the embryo against pests and pathogens. In this work the vicilin from Anadenanthera macrocarpa - AmV (red-angico), was purified and partially characterized, its effect on development and larval survival and adult emergence of Callosobruchus maculatus was evaluated by determination of LD50, WD50 and ED50 in system bioassay. Purification of vicilin was initiated by the chitin affinity chromatography and then gel filtration (Superdex 75 Tricorn 10x300 mm) FPLC system followed by reverse phase chromatography (C8 phenomenex) on HPLC system. Bioassays WD50 and LD50 for larvae were 0.32% and 0.33% (w:w) respectively, since the ED50 for adults was 0.096%. The probable mechanism of action was evaluated by testing digestibility of AmV in vitro, and observed for the involvement of two fragments vicilins immunoreactive against polyclonal Anti-vicilin from Erythrina velutina (Anti-EvV) about of 22 and 13 kDa chitin binding. The AmV in its native form has been recognized by the anti-EvV, indicating that there is a conserved region in the vicilin and is probably corresponding to the chitin binding domains. These results point to a new vicilin chitin binding that can subsequently be used as a possible biopesticide protein source, in order to control insect pest C. maculatus and confirm literature findings that demonstrate vicilin in the presence of different kinds of ligands to conserved regions chitin not yet characterized

Spatial variability of soil properties and yield of a grazed alfalfa pasture in Brazil.

2016

A pigeonpea gene confers resistance to Asian soybean rust in soybean.

Asian soybean rust (ASR), caused by the fungus Phakopsora pachyrhizi, is one of the most economically important crop diseases, but is only treatable with fungicides, which are becoming less effective owing to the emergence of fungicide resistance. There are no commercial soybean cultivars with durable resistance to P. pachyrhizi, and although soybean resistance loci have been mapped, no resistance genes have been cloned. We report the cloning of a P. pachyrhizi resistance gene CcRpp1 (Cajanus cajan Resistance against Phakopsora pachyrhizi 1) from pigeonpea (Cajanus cajan) and show that CcRpp1 confers full resistance to P. pachyrhizi in soybean. Our findings show that legume species related to soybean such as pigeonpea, cowpea, common bean and others could provide a valuable and diverse pool of resistance traits for crop improvement.

Aluminium tolerance in bean traditional cultivars from Madeira

Common bean (Phaseolus vulgaris L.) is the most important legume crop in the world, providing low-cost, high quality protein, minerals and dietary fiber for human nutrition. The crop was originated from diversity centers in America and exhibits adaptation abilities to different environmental conditions, including soil with low pH. Acid soils occupy 30% of the agro ecosystem areas in the world. In Madeira, acid Andosols and unsatured Cambisols are the dominant groups of soils. Generally, under acidic and infertile conditions, besides of H+ toxicity, soluble aluminium (Al) is the most important abiotic factor limiting plant development and crop productivity. In the field, the hidden roots are also affected and the reduction of root growth under Al stress can be clearly observed in early stages. Seedlings of fifty bean accessions from the Archipelago of Madeira were tested under controlled conditions in the presence of 50 mM Al at pH 4.4. In general, the tested germplasm appeared to be sensitive or very sensitive to Al toxicity. However, fifteen traditional cultivars clearly exhibited elevated Al-tolerance, with an average root relative elongation (RRE) exceeding 50%, while top six accessions surpassed the 60% RRE mark. The Madeira bean germplasm is a valuable resource for sustainable crop production in acid soils and it could be used as parental lines in breeding programs aimed for Al tolerance in common beans.

Aluminium tolerance in bean traditional cultivars from Madeira

Common bean (Phaseolus vulgaris L.) is the most important legume crop in the world, providing low-cost, high quality protein, minerals and dietary fiber for human nutrition. The crop was originated from diversity centers in America and exhibits adaptation abilities to different environmental conditions, including soil with low pH. Acid soils occupy 30% of the agro ecosystem areas in the world. In Madeira, acid Andosols and unsatured Cambisols are the dominant groups of soils. Generally, under acidic and infertile conditions, besides of H+ toxicity, soluble aluminium (Al) is the most important abiotic factor limiting plant development and crop productivity. In the field, the hidden roots are also affected and the reduction of root growth under Al stress can be clearly observed in early stages. Seedlings of fifty bean accessions from the Archipelago of Madeira were tested under controlled conditions in the presence of 50 mM Al at pH 4.4. In general, the tested germplasm appeared to be sensitive or very sensitive to Al toxicity. However, fifteen traditional cultivars clearly exhibited elevated Al-tolerance, with an average root relative elongation (RRE) exceeding 50%, while top six accessions surpassed the 60% RRE mark. The Madeira bean germplasm is a valuable resource for sustainable crop production in acid soils and it could be used as parental lines in breeding programs aimed for Al tolerance in common beans.

Heat shock response in bacteria with large genomes: lessons from rhizobia

Rhizobia are important soil bacteria due to their ability to establish nitrogen-fixing symbioses with legume plants. In this dual lifestyle, as free-living bacteria or as plant symbiont, rhizobia are often exposed to different environmental stresses. The present chapter overviews the current knowledge on the heat shock response of rhizobia, highlighting how these large genome bacteria respond to heat from a transcriptional point of view. Response to heat shock in rhizobia involves genome wide changes in the transcriptome that may affect more than 30% of the genome and involve all replicons. In addition to the expected upregulation of genes already known to be involved in stress response (dnaK, groEL, ibpA, clpB), the reports on the heat shock response in rhizobia also showed particular aspects of stress response in these resourceful bacteria. The transcriptional response to heat in rhizobia includes the overexpression of a large number of genes involved in transcription and carbohydrate transport and metabolism. Additional studies are needed in order to better understand the transcriptional regulation of stress response in bacteria with large genomes.

Stylosanthes guianensis cv. BRS Bela managed as organic fodder bank for supplementation of lactation crossbred cows in the Cerrado.

2016

Organic management of Brachiaria brizantha cv. Marandu pasture mixed with Stylosanthes spp. in the Cerrado.

2016

Beyond conservation agriculture.

ABSTRACT: Global support for Conservation Agriculture (CA) as a pathway to Sustainable Intensification is strong. CA revolves around three principles: no-till (or minimal soil disturbance), soil cover, and crop rotation. The benefits arising from the ease of crop management, energy/cost/time savings, and soil and water conservation led to widespread adoption of CA, particularly on large farms in the Americas and Australia, where farmers harness the tools of modern science: highly-sophisticated machines, potent agrochemicals, and biotechnology. Over the past 10 years CA has been promoted among smallholder farmers in the (sub-) tropics, often with disappointing results. Growing evidence challenges the claims that CA increases crop yields and builds-up soil carbon although increased stability of crop yields in dry climates is evident. Our analyses suggest pragmatic adoption on larger mechanized farms, and limited uptake of CA by smallholder farmers in developing countries. We propose a rigorous, context-sensitive approach based on Systems Agronomy to analyze and explore sustainable intensification options, including the potential of CA. There is an urgent need to move beyond dogma and prescriptive approaches to provide soil and crop management options for farmers to enable the Sustainable Intensification of agriculture.

Agronomic efficiency of Bradyrhizobium in peanut under different environments in Brazilian Northeast.

Several legumes have natural ability to associate with nitrogen - fixing bacteria known as rhizobia. The efficiency of this association depends on the plant and bacterial genotype and the edaphoclimatic conditions. Peanut is a tropical legume able to associate with a wide range of rhizobia and the selection of efficient bacteria is important to increase the nitrogen fixation in this crop. In order to investigate the agronomic efficiency of two Bradyrhizobiumstrains, two peanut genotypes were used in field trails carried out in three environments located at Brazilian Northeast. The genotypes (BR1 and L7 Bege) were submitted to rhizobial inoculation (SEMIA 6144 or ESA 123, both Bradyrhizobium strains, and chemical nitrogen fertilization in randomized block design experiments. The following traits were analyzed: flowering (F), main axis height (MAH), number of nodules/plant (NN), number of pods/plant (NP) and weight of pods (WP). Differential responses were found in all to treatments to NN, NP and WP, in the three environments studied. Overall, ESA 123 showed good agronomic performance inducing higher pod production. The results supportthe evaluation of the Bradyrhizobium in further experiments aiming at its recommendation to commercial inoculants in Brazilian Northeast region.

Forage peanut (Arachis spp.) genetic evaluation and selection.

Forage peanut improvement for use in grass?legume mixtures is expected to have a great impact on the sustainability of Brazilian livestock production. Eighteen cloned Arachis spp. ecotypes were evaluated under clipping in a Brazilian Cerrado region and results analysed using a mixed model methodology. The objective was to estimate genetic and phenotypic parameters and to select the best ecotypes based on selection index applied on their predicted genotypic value. The traits of total dry-matter (DM) and leaf DM yield presented moderate (0_30 < h2g < 0_50) to high (>0_50) broad-sense heritability, in contrast to the low genetic variability in nutritional quality-associated traits. Ecotypes of Arachis spp. contained average crude protein concentrations of 224 g kg _1 DM in leaves and 138 g kg _1 DM in stems, supporting the potential role of these species to overcome the low protein content in Cerrado pastures. The correlations between yield traits and traits associated with low nutritional value in leaves were consistently significant and positive. Genetic correlations among all the yield traits evaluated during the rainy or dry seasons were significant and positive. The ecotypes were ranked based on selection index. The next step is to validate long-term selection of grass?Arachis in combination with pastures under competition and adjusted grazing in the Cerrado region.

Seleção de marcadores microssatélites e otimização de protocolo de amplificação em alfafa.

2016

Endemic Mimosa species from Mexico prefer alphaproteobacterial rhizobial symbionts.

2016

Innovative strategies for the mitigation of acrylamide content in different food products

Acrylamide (AA) is an undesirable food toxic compound, classified as 'probably carcinogenic to humans' by the International Agency for Research on Cancer due to its toxic effects, including neurotoxicity, genotoxicity, carcinogenicity and reproductive toxicity. AA is mainly formed during the heat treatment of foods (> 120 °C) by the Maillard reaction, an essential reaction that also allows the desired levels of shelf-life and sensory properties of various food products to be achieved. Over the years, authorities and regulations have become more restrictive regarding the maximum levels of AA permitted in foods and beverages. The latest Commission Regulation (EU) 2017/2158 contains reference levels and measures to reduce AA in several food groups that contribute to the highest dietary intake, making necessary the study of promising AA mitigation strategies. The aim of this PhD research project was to identify, characterise and optimise some AA mitigation strategies in the most at-risk widely consumed foods such as potato, coffee and bakery products. Some AA control strategies were selected and investigated for each food category, also considering the main quality characteristics of the final products. The comprehensive results obtained during the three years of research activity have allowed a deeper knowledge of the traditional and innovative AA mitigation strategies, which can be extremely useful for both the food industry and international authorities. The most promising strategies studied in terms of reduction of AA while maintaining the main quality characteristics of the examined foods were: the application of pulsed electric fields and yeast immersion as pre-treatments of chips for frying; the selection of high roasting degrees for coffee products; the selection of static baking conditions for biscuits; the optimisation of alternative biscuit’ formulations by both the use of chickpea legume flour and of flour from bean with intact cotyledon cell walls.