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

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

Avaliação da ação bioinseticida de SBTI e vicilina de Erythrina velutina em enzimas digestivas e membrana peritrófica de larvas de Plodia interpunctella (Lepidoptera: Pyralidae)

Plodia interpunctella (Indian meal moth) is a cosmopolitan pest that attacks not only a wide range of stored grain as well other food products. Due to its economic importance several researches have focused in a method with ability to control this pest with few or no damage to the environment. The study of digestive enzymes inhibitors, lectins and chitin-binding proteins, has often been proposed as an alternative to reduce insect damage. In this study we report the major classes of digestive enzymes during larval growth in P. Interpunctella, being those proteinases actives at pH 9.5 and optimum temperature of 50 oC to both larvae of the 3rd instar and pre-pupal stage of development. In vitro and zymogram assays presented the effects of several inhibitors, such as SBTI, TLCK and PMSF to intestinal homogenate of 3rd instar larvae of 62%, 92% and 87% of inhibition and In pre-pupal stage of 87%, 62 % and 55% of inhibition, respectively. Zymograms showed inhibition of two low molecular masses protein bands by TLCK and that in presence of SBTI were retarded. These results are indicative of predominance of digestive serine proteinases in gut homogenate from Plodia interpunctella larvae. This serine proteinase was then used as a target to evaluate the effect of SBTI on larvae in in vivo assay. Effect of SBTI on mortality and larval mass was not observed at until 4% of concentration (w/w) in diets. Chitin, another target to insecticidal proteins, was observed by chemical method. Moreover, optic microscopy confirmed the presence of a peritrophic membrane. Established this target, in vivo effect of EvV, a chitin binding vicilin, evaluated during the larval development of P. interpunctella and was obtained a LD50 of 0,23% and WD50 of 0,27% to this protein. Mechanism of action was proposed through of the in vivo digestibility of EvV methodology. During the passage through the larval digestive tract was observed that EvV was susceptible to digestive enzymes and a reactive fragment, visualized by Western blotting, produced by digestion was recovered after dissociation of the peritrophic membrane. The bound of EvV to peritrophic membrane was confirmed by immunohystochemical assays that showed strong immunofluorescent signal of EvV-FITC binding and peritrophic membrane. These results are a indicative that vicilins could be utilized as potential insecticide to Plodia interpunctella and a control methods using EvV as bioinsecticide should be studied to reduce lost caused by storage insect pests

Interação de microorganismos na solubilização de fósforo e potássio de rochas para produção de biofertilizantes

he present model of agriculture is based on intensive use of industrial inputs, due to its rapid response, but it brings harmful consequences to the environment, and it is necessary the use of modern inputs. And an alternative is the use of rock biofertilizers in agriculture, a product easy to use, with higher residual effect and does not harm the environment. The objective of study was to evaluate the inoculation and co-inoculation of different microorganisms in the solubilization of rock phosphate and potash ground microbial evaluating the best performance in the production of biofertilizers comparing with rocks pure in soil chemical properties and, verify effect of inoculation of the bacterium Paenibacillus polymyxa in the absorption of minerals dissolved in the development of cowpea (Vigna unguiculata [L.] Walp.). The first bioassay was conducted in Laboratory (UFRN) for 72 days in Petri dishes, where the rock powder was increased by 10% and sulfur co-inoculated and inoculated with bacterial suspension of Paenibacillus polymyxa grown in medium tryptone soy broth, Ralstonia solanacearum in medium Kelman, Cromobacterium violaceum in medium Luria-Bertani and Acidithiobacillus thiooxidans in medium Tuovinen and Kelly,and fungi Trichoderma humatum and Penicillium fellutanum in malt extract. Every 12 days, samples were removed in order to build up the release curve of minerals. The second bioassay was conducted in a greenhouse of the Agricultural Research Corporation of Rio Grande do Norte in experimental delineation in randomized block designs, was used 10 kg of an Yellow Argissolo Dystrophic per pot with the addition of treatments super phosphate simple (SS), potassium chloride (KCl), pure rock, biofertilizers in doses 40, 70, 100 and 200% of the recommendation for SS and KCl, and a control, or not inoculated with bacteria P. polymyxa. Were used seeds of cowpea BRS Potiguar and co-inoculated with the bacterial suspension of Bradyrhizobium japonicum and P. polymyxa. The first crop was harvested 45 days after planting, were evaluated in the dry matter (ADM), macronutrients (N, P, K, Ca, Mg) and micronutrients (Zn, Fe, Mn) in ADM. And the second at 75 days assessing levels of macro end micronutrients in plants and soil, and the maximum adsorption capacity of P in soil. The results showed synergism in co-inoculations with P. polymyxa+R. solanacearum and, P. polymyxa+C. violaceum solubilizations providing higher P and K, respectively, and better solubilization time at 36 days. The pH was lower in biofertilizers higher doses, but there was better with their addition to P at the highest dose. Significant reduction of maximum adsorption capacity of phosphorus with increasing dose of biofertilizer. For K and Ca was better with SS+KCl, and Mg to pure rock. There was an effect of fertilization on the absorption, with better results for P, K and ADM with SS+KCL, and N, Ca and Mg for biofertilizers. Generally, the P. polymyxa not influence the absorption of the elements in the plant. In treatments with the uninoculated P. polymyxa chemical fertilizer had an average significantly higher for weight and number of grains. And in the presence of the bacteria, biofertilizers and chemical fertilizers had positive values in relation to rock and control. The data show that the rocks and biofertilizers could meet the need of nutrients the plants revealed as potential for sustainable agriculture

Allelopathic plants. 7. Sunflower (Helianthus annuus L.)

Sunflower is an annual dicotyledonous plant, herbaceous, erect and native of North America. It is thermo- and photo-insensitive, hence, can be grown round the year in sub-tropical and tropical countries. Only two spp. H. annuus and H. tuberosum are cultivated for food, remaining spp. are ornamentals, weeds and wild plants. However, H. annuus is allelopathic and inhibit the growth and development of other plants thus reducing their productivity. Much information is available about the allelopathic effects of sunflower crop on following crops in crop rotations. Although it is harmful to all crops, but, is less harmful to crops of Graminae family than other families. It seems that the harmful effects of sunflower in crop rotations are due to release and accumulation of root exudates during crop growth in soil. Soil incorporation of its fresh (green manure) or dry biomass in soil is inhibitory to both crops and weed spp. Several allelochemicals have been characterized from the H. annuus, which inhibit the seed germination and seedling growth of A. albus, A. viridis, Agropyron repens (Elymus repens), Ambrosia artemsiifolia, Avena fatua, Celosia crustata, Chenopodium album, Chloris barbara, Cynodon dactylon, D. sanguinalis, Dactyloctenium ageyptium, Digitaria ciliaris, Echinochloa crus-galli, Flaveria australasica, Parthenium hysterophorus, Portulaca oleracea, Sida spinosa, Trianthema portulacastrum, Veronica perisca the inhibitory effects of this crop may be used for weed management with less herbicides for sustainable agriculture.

Patogenicidade in vitro de Monacrosporium robustum a Rotylenchulus reniformis

O nematóide reniforme (Rotylenchulus reniformis) é um importante patógeno de importantes culturas brasileiras, tais como soja, algodão, feijão, caupi, maracujazeiro, mamoeiro, tomate e alface, entre outras. Dentre os fungos nematófagos com potencial de utilização comercial como agentes do controle biológico de nematóides, destacam-se espécies do gênero Monacrosporium. No presente estudo, avaliou-se, in vitro, a capacidade predatória e de parasitismo de Monacrosporium robustum ao nematóide reniforme. em períodos de 24, 48 e 72 horas após, constatou-se que a percentagem de predação do nematóide pelo fungo foi de, respectivamente, 81,3, 99,9 e 100.

Não preferência para alimentação e aspectos biológicos de Spodoptera eridania em cultivares de feijão-caupi

The aim of this paper was to evaluate the no-preference for feeding and biological aspects of Spodoptera eridania fed on cowpea cultivars BR17 Gurgueia, BRS Urubuquara, BRS Nova Era, Sempre Verde, BRS Milenio and BR3 Tracuateua. In free-choice test, leaf discs were placed in Petri dishes where one third instar larvae per cultivar was released, whereas in no-choice test one leaf disc was placed per Petri dish where one caterpillar per cultivar was released, evaluating their attractiveness after 1, 3, 5, 10, 15, 30, 60, 120, 360 and 720 minutes, as well as the leaf area consumed. Randomized blocks and complete randomized blocks design were used for free-choice and no-choice tests, respectively, with six treatments and 10 replications. The evaluation of the biological parameters of S. eridania was carried out in Petri dishes where recently hatched caterpillars were transferred in the proportion of one per dish, and the leafs of the cultivars were offered to them during the whole larval period, and we evaluated: periods and viabilities of larvae and pupae, overall viability, weight of larvae and pupae, sex ratio, longevity and overall cycle. Complete randomized design was used with six treatments and 30 replications. In no- preference for feeding free-choice test the cultivars Sempre Verde and BR17 Gurgueia were the most and the least consumed, respectively. Regarding the effects of cowpea cultivars on larval viability we can infer that BRS Urubuquara and Sempre Verde show antibiosis-type resistance to S. eridania.

Efeito de genótipos de caupi, Vigna unguiculata (L.) Walp., sobre o desenvolvimento de Callosobruchus maculatus (Fabricius) (Coleoptera: Bruchidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Atratividade de genótipos de feijão-caupi para oviposição de Bemisia tabaci biótipo B

O objetivo deste trabalho foi avaliar a atratividade de genótipos de feijão-caupi para oviposição de Bemisia tabaci biótipo B e identificar possíveis fontes de resistência à mosca-branca. Foram avaliados 51 genótipos, com uso de testes de chance de escolha. Os genótipos foram divididos aleatoriamente em dois grupos, tendo-se utilizado o genótipo Canapu como testemunha sucetível. Os 14 genótipos mais promissores (sete de cada grupo) foram selecionados para a realização de ensaios complementares (com ou sem chance de escolha). No teste com chance de escolha, os genótipos BRS-Urubuquara, TVU-36, TE93-244-23 F-1, BR 17-Gurgueia, BRS-Marataoã, MNC99-541 F-21 e TE97-304 G-4 foram menos atrativos à mosca-branca. Os genótipos TE93-244-23 F-1 e TVU-36 apresentaram resistência pelo mecanismo de não preferência para ovoposição. No teste sem chance de escolha, apenas o genótipo TVU-36 apresentou resistência por esse mecanismo.

Incidência e severidade do vírus do endurecimento dos frutos em maracujazeiros enxertados em pé-franco

O objetivo deste trabalho foi avaliar a incidência e a severidade do vírus do endurecimento dos frutos em maracujazeiro-amarelo enxertado e pé-franco. O experimento foi conduzido no município de Adamantina-SP, no período de abril de 2006 a junho de 2007, adotando-se o delineamento em blocos ao acaso, com quatro tratamentos e oito repetições. Foram avaliados três porta-enxertos: Passiflora edulis, P. alata e P. gibertii, e plantas de pé-franco. Utilizou-se como copa o maracujazeiro-amarelo (Passiflora edulis Sims). Avaliaram-se a porcentagem de plantas com sintomas de virose e a severidade dos sintomas. As primeiras plantas com sintomas de virose ocorreram aos 90 dias do plantio das mudas no campo, atingindo, aos 180 dias, 100% de plantas com virose em P. alata e P. gibertii, e 97,5% em P. edulis e pé-franco.

Práticas agroecológicas para o cultivo do gergelim (Sesamum indicum L.)

The exceeded use of the natural resources required by the modern agriculture has been caused soil impoverishment, soil salinization and soil compaction. The unreasonable use of chemical fertilizers and pesticides causes chemical imbalances in the plant tissues, nutritional losses, taste chances and human health problems. The monocrops are more vulnerable to the pest and disease attacks. The aim of this work were to indicate the better relative planting time of the cowpea bean for the sesame/cowpea bean intercropping based on the evaluation of agronomic, economic, ecological and physiological parameters and to evaluate the viability of the cow urine and cassava wastewater use as alternative fertilizers on the sesame cultivation. In a field essay, when the sesame/cowpea bean was evaluate, the mainly treatments were the single planting of the two cultures and the multiple cropping, with the cowpea bean being planted on the same time, 7, 14 and 21 days after the sesame. The mainly treatments of the second essay were the combined doses of the cow urine (0 and 50 ml) and cassava wastewater (0, 500 and 1000 mL). In the intercrop evaluation significant differences were found between the treatments for the most of the parameters. It was verified that as the sowing of the cowpea bean was retarded in relation to the sesame sowing, the sesame performance increased when the cowpea bean performance decreased. The treatment 7 days balanced the competition relations that occurred between the two crops in the intercrop. The sesame presented little pronounced responses to the different doses of cassava wastewater. The cow urine affected the sesame growth and the growth rates increased with the increasing of the application of the product. The responses of the sesame growth to the cow urine application must be related both the nutritional richness of the product and the presence of growth stimulating substances. The obtained results indicated that in the sesame/cowpea bean intercropping, when the sesame is planted 7 days after the sesame, there is a higher possibility of the net gains to the farmer. Cow urine and cassava wastewater may be used as additional organic fertilizers

Expressão diferencial dos genes VuUCP1a e VuUCP1b em caupi sob estresse salino

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Photosynthetic limitations caused by different rates of water-deficit induction in Glycine max and Vigna unguiculata

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Identification and characterization of a potyvirus isolated from siratro plants

The present work describes the identification and characterization of a potyvirus isolated from siratro (Macroptilium atropurpureum Urb.) in the north-west region of the State of Sdo Paulo, Brazil. The virus was transmitted by mechanical inoculation. Its host range was restricted mainly to members of the Fabaceae. A cDNA fragment of about 930 bp was amplified by RT/PCR, cloned and sequenced. The fragment, which included the coat protein gene, had amino acid identity percentages between 88 and 98% with isolates of Bean common mosaic virus (BCMV). Phylogenetic analysis grouped the. siratro potyvirus and BCMV isolates in 99% of the replicates, including Azuki mosaic virus, Dendrobium mosaic virus, Blackeye cowpea mosaic virus and Peanut stripe virus, which have been classified as BCMV strains. This is the first citation on the presence of BCMV in siratro plants in Brazil.

Detection and partial characterization of a carlavirus causing stem necrosis of soybean in Brazil

No ano agrícola de 2000/2001, ocorreu um surto de nanismo e necrose da haste em soja (Glycine max) plantada em duas áreas do Brasil Central e na safra seguinte, esta anomalia foi constatada em outras regiões produtoras, mesmo distantes mais de 2.000 km de onde fora inicialmente constatada. Estudos envolvendo ensaios de transmissão (enxertia, mecânica e insetos vetores), microscopia eletrônica, purificação, sorologia e ensaios moleculares indicaram que a enfermidade foi causada por um carlavirus transmitido por mosca branca, possivelmente relacionado ao Cowpea mild mottle virus (CpMMV).

Variabilidade e correlações entre caracteres agronômicos em caupi (Vigna unguiculata)

O caupi (Vigna unguiculata (L.) Walp) é um alimento básico das populações do Nordeste brasileiro, devendo merecer atenção com vistas a melhoria da qualidade de grãos, resistência a doenças e pragas e aumento de produtividade. Este trabalho teve por objetivo estudar a variabilidade e o potencial genético de 28 linhagens, escolhidas após uma seleção para cor, tamanho de grãos e resistência a viroses. A produtividade apresentou coeficiente de variação genético de 23,90%, e o valor agronômico, de 3,56%. O número de vagens por pedúnculo apresentou a menor estimativa do coeficiente de determinação genético (4,51%), e o peso de 100 grãos, a maior (81,74%). O coeficiente de determinação genético da produtividade foi de 34,15%. As maiores estimativas de ganho genético foram as do peso de 100 grãos (21,73%) e da produtividade (19,77%). As correlações genotípicas foram superiores às fenotípicas e às de ambiente, destacando-se as correlações entre número de ramos secundários e produtividade (68,13%), e valor agronômico e produtividade (100%). Estes resultados mostram amplas possibilidades de seleção entre as linhagens com relação à maioria dos caracteres estudados.