Legume type and temperature effects on the toxicity of insecticide to the genus Callosobruchus (Coleoptera: Bruchidae)

Three bruchid pest species, Callosobruchus maculatus, Callosobruchus chinensis and Callosobruchus rhodesianus, were studied for their response to insecticide toxicity taking into account the separate and interactive effects of temperature and pre-adult food. The food types used were cowpea (Vigna unguiculata) and mungbean (Vigna radiata). Callosobruchus maculatus was the most tolerant to malathion and the least affected by temperature change while C. rhodesianus was the least tolerant. Over a 4 C range (23, 25, 27 C), there was generally a significant impact of temperature on the tolerance of the three species to the insecticide. The food type on which the insects developed influenced considerably the degree of insecticide tolerance. Callosobruchus maculatus and C. chinensis populations reared onmungbean had higher tolerance to malathion than their counterparts reared on cowpea, but the opposite was observed in C. rhodesianus populations. The food influence in this study suggested an ancestral cause or fitness cost depending on the species. The interaction of food-by-temperature had no significant effect on malathion toxicity to this genus. Correlation analysis showed C. chinensis to be relatively less sensitive to insecticide concentration over the range studied compared with the other two species.

Interactive effect of cowpea variety, dose and exposure time on bruchid tolerance to botanical pesticides

Callosobruchus maculatus has for years remained a serious menace in cowpea in Sub-Sahara Africa. The objective of this study was to investigate the effect of genotypic cowpea (Vigna unguiculata (L.) Walp) varieties, time and dose on C. maculatus exposed to powders of Piper guineense and Eugenia aromatica. Irrespective of duration and botanicals, bruchid reared on KDV showed the highest tolerance to both plant materials; while their counterparts from IAR48V were the most susceptible. Median lethal time (LT50) also varied according to the plant materials; with the highest in KDV reared bruchid [P. guineense: KDV (18.31), IAR48V (9.27), IFBV (13.17); E. aromatica: KDV (76.01), IAR48V (5.59), IFBV (6.49)]. There was a significant impact of cowpea variety (V), exposure time (T) and dose (D) on the tolerance of C. maculatus to both plant materials. The effect of all two-way (VxT, VxD, DxT) and three way interactions (V×T×D) on the tolerance of C. maculatus to both plant materials was also significant. Varietal effect was more pronounced in bruchids exposed to E. aromatica; while exposure time was more pronounced in bruchids exposed to P. guineense.

HOST-MEDIATED INDUCTION OF alpha-AMYLASES BY LARVAE OF THE MEXICAN BEAN WEEVIL Zabrotes subfasciatus (COLEOPTERA: CHRYSOMELIDAE: BRUCHINAE) IS IRREVERSIBLE AND OBSERVED FROM THE INITIATION OF THE FEEDING PERIOD

Larvae of Zabrotes subfasciatus secrete alpha-amylases that are insensitive to the alpha-amylase inhibitor found in seeds of Phaseolus vulgaris. By analyzing amylase activities during larval development on P. vulgaris, we detected activity of the constitutive amylase and the two inducible amylase isoforms at all stages. When larvae were transferred from the non alpha-amylase inhibitor containing seeds of Vigna unguiculata to P. vulgaris, the inducible alpha-amylases were expressed at the same level as in control larvae fed on P. vulgaris. Interestingly, when larvae were transferred from seeds of P. vulgaris to those of V. unguiculata, inducible alpha-amylases continued to be expressed at a level similar to that found in control larvae fed P. vulgaris continuously. When 10-day-old larvae were removed from seeds of V. unguiculata and transferred into capsules containing flour of P. vulgaris cotyledons, and thus maintained until completing 17 days ( age when the larvae stopped feeding), we could detect higher activity of the inducible alpha-amylases. However, when larvae of the same age were transferred from P. vulgaris into capsules containing flour of V. unguiculata, the inducible alpha-amylases remained up-regulated. These results suggest that the larvae of Z. subfasciatus have the ability to induce insensitive amylases early in their development. A short period of feeding on P. vulgaris cotyledon flour was sufficient to irreversibly induce the inducible alpha-amylase isoforms. Incubations of brush border membrane vesicles with the alpha-amylase inhibitor 1 from P. vulgaris suggest that the inhibitor is recognized by putative receptors found in the midgut microvillar membranes. (C) 2010 Wiley Periodicals, Inc.

Characterisation, immunolocalisation and antifungal activity of a lipid transfer protein from chili pepper (Capsicum annuum) seeds with novel alpha-amylase inhibitory properties

Lipid transfer proteins (LTPs) were thus named because they facilitate the transfer of lipids between membranes in vitro. This study was triggered by the characterization of a 9-kDa LTP from Capsicum annuum seeds that we call Ca-LTP(1). Ca-LTP(1) was repurified, and in the last chromatographic purification step, propanol was used as the solvent in place of acetonitrile to maintain the protein`s biological activity. Bidimensional electrophoresis of the 9-kDa band, which corresponds to the purified Ca-LTP(1), showed the presence of three isoforms with isoelectric points (pIs) of 6.0, 8.5 and 9.5. Circular dichroism (CD) analysis suggested a predominance of alpha-helices, as expected for the structure of an LTP family member. LTPs immunorelated to Ca-LTP(1) from C. annuum were also detected by western blotting in exudates released from C. annuum seeds and also in other Capsicum species. The tissue and subcellular localization of Ca-LTP(1) indicated that it was mainly localized within dense vesicles. In addition, isolated Ca-LTP(1) exhibited antifungal activity against Colletotrichum lindemunthianum, and especially against Candida tropicalis, causing several morphological changes to the cells including the formation of pseudohyphae. Ca-LTP(1) also caused the yeast plasma membrane to be permeable to the dye SYTOX green, as verified by fluorescence microscopy. We also found that Ca-LTP(1) is able to inhibit mammalian alpha-amylase activity in vitro.

The defensive functions of plant inhibitors are not restricted to insect enzyme inhibition

Three plant proteinase inhibitors BbKI (kallikrein inhibitor) and BbCI (cruzipain inhibitor) from Bauhinia bouhinioides, and a BrTI (trypsin inhibitor) from B. rufa, were examined for other effects in Callosobruchus maculatus development; of these only BrTI affected bruchid emergence. BrTI and BbKI share 81% identities in their primary sequences and the major differences between them are the regions comprising the RGD and RGE motifs in BrTI. These sequences were shown to be essential for BrTI insecticidal activity, since a modified BbKI [that is a recombinant form (BbKIm) with some amino acid residues replaced by those found in BrTI sequence] also strongly inhibited insect development. By using synthetic peptides related to the BrTI sequence, YLEAPVARGDGGLA-NH(2) (RGE) and IVYYPDRGETGL-NH(2) (RGE), it was found that the peptide with an RGE sequence was able to block normal development of C. maculatus larvae (ED(50) 0.16% and LD(50) 0.09%), this being even more effective than the native protein. (C) 2009 Elsevier Ltd. All rights reserved.

Coordenação entre as respostas da catalase e da ascorbato peroxidade em folhas de feijão caupi submetidas a diferentes fontes de peróxido de hidrogênio

The plants are often exposed to variations in environmental conditions that may trigger metabolic disturbances leading to a consequent loss in productivity of crops. These stressful conditions usually induce an accumulation of reactive oxygen species (ROS) in the cell, a condition known how oxidative stress. Among these species, hydrogen peroxide (H2O2) is an important molecule involved in numerous signaling mechanisms. The present study aimed to understand the relationship between the different enzymatic mechanisms of elimination of H2O2 by catalase (CAT) and ascorbate peroxidase (APX) in leaf tissues of seedlings of the species Vigna unguiculata L. Walp, under conditions of oxidative stress induced by application of CAT inhibitor, 3-amino-1,2,4-triazole (3-AT), and H2O2 itself on the roots. Three experiments were conducted. The first experiment was performed applying the compound 3-AT (5 mM) during the time (hours). In the second experiment, seedlings were exposed to different concentrations of H2O2 (2.5, 5.0, 7.5, 10 mM) for 48 h. The third strategy included the pre-treatment with H2O2 (2.5 mM) for 24 h, followed by subsequent treatment with the inhibitor 3-AT and recovery control condition. Treatment with 3-AT causes a strong inhibition of CAT activity in leaf tissues accompanied by an increase of activity of APX. However a decrease in oxidative damage to lipids is not observed as indicated by TBARS. It was observed that activity of APX is directly linked to the content of peroxide. Inductions in the activities of CAT and APX were observed mainly in the seedlings treated with 2.5 mM H2O2. This can be associated with a decrease in oxidative damage to lipids. In contrast, one same tendency was not observed in treatments with higher concentrations of this ROS. These results suggest that the concentration of 2.5 mM H2O2 can induce responses antioxidants later in seedling cowpea. This concentration when applied as pre-treatment for 24 h promoted an induction systems removers CAT and APX, both in activity and in terms of gene expression. However this increment was not observed in the recovered plants and the plants subsequently subjected to 3-AT. Additionally, the pretreatment was not sufficient to attenuate the inhibition of CAT activity and oxidative damage to lipids caused by the subsequent application of this inhibitor. The results showed that the application of 3-AT and H2O2 in the root systems of seedlings of cowpea promote changes in the parameters analyzed in leaf tissues that indicate a direct response to the presence of these factors or systemic signaling mecanisms. H2O2 appears to activate the responses of two antioxidant systems in this study thar does not promote greater protection in case of additional treatment with 3-AT. This demonstrates the importance of the CAT system. In this work, complete results indicate that there is a difference between the signaling and the effects caused by exposure to H2O2 and by treatment with 3-AT

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

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.

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.

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

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Dinâmica de populações e fitossociologia de plantas daninhas no cultivo do feijão-caupi e mandioca no sistema corte e queima com o uso de arado

Este estudo investigou a composição florística das plantas daninhas em área queimada durante três anos agrícolas. A pesquisa foi conduzida no município de Zé Doca, Maranhão. O preparo da área no primeiro ano agrícola (2006/2007) foi realizado com corte e queima da vegetação para o cultivo de milho seguido do feijão-caupi. No segundo e no terceiro ano agrícola, o preparo da área consistiu de aração para o cultivo do milho seguido de mandioca (2007/2008) e depois para o feijão-caupi em sucessão à cultura de mandioca (2008/2009). A coleta das plantas daninhas ocorreu nas culturas de feijão-caupi e mandioca aos 30 e 60 dias após a semeadura (DAS), no primeiro e no segundo ano agrícola, respectivamente, e no feijão-caupi aos 30 DAS do terceiro ano agrícola, com retângulo (0,5 x 0,3 m) lançado 10 vezes ao acaso na área cultivada. A cada lançamento, as plantas daninhas foram colhidas, para contagem, identificação, secagem e, assim, obter os índices fitossociológicos. O fogo reduziu a diversidade e o número das plantas daninhas. As espécies com maior valor de IVI foram Imperata brasiliensis, Sida glomerata e Corchorus argutus, após o fogo na cultura do feijãocaupi; e Juncus sp., Spermacoce verticillata, Aeschynomene americana e Cyperus sp., após preparo da área com aração nas culturas de mandioca e feijão-caupi. As plantas de capoeira ocorreram depois da queima, porém sua importância foi reduzida com o passar do tempo.

Preliminary study of a methodology based on the degree-days concept for predicting the phenological phases of crops

This work develops a methodology (using the degree-days concept and linear regression), to forecast the duration of phenological phases in crops. An experiment was conducted in the greenhouse with three cultivars of cowpea (Vigna unguiculata (C.) Walp.), cv. California-781, Tvx 5058-09C and IT 81D-1032. The methodology was based on the relative thermal efficiency rate, determined for each species or cv. The results show that the proposed methodology may be a good alternative in works involving crops, especially because it does not require the repetition of the experiments.

Estudo preliminar de uma metodologia baseada no conceito de graus-dia, para previsão das fases fenológicas de culturas agrícolas

This work develops a methodology (using the degree-days concept and linear regression), to forecast the duration of phenological phases in crops. An experiment was conducted in the greenhouse with three cultivars of cowpea (Vigna unguiculata (L.) Walp.), cv. Califórnia-781, Tvx 5058-09C and IT 81D-1032. The methodology was based on the relative thermal efficiency rate, determined for each species or cv. The results show that the proposed methodology may be a good alternative in works involving crops, especially because it does not require the repetition of the experiments.

Identification of duplicated and stress-inducible Aox2b gene co-expressed with Aox1 in species of the Medicago genus reveals a regulation linked to gene rearrangement in leguminous genomes

In flowering plants, alternative oxidase (Aox) is encoded by 3-5 genes distributed in 2 subfamilies (Aox1 and Aox2). In several species only Aox1 is reported as a stress-responsive gene, but in the leguminous Vigna unguiculata Aox2b is also induced by stress. In this work we investigated the Aox genes from two leguminous species of the Medicago genus (Medicago sativa and Medicago truncatula) which present one Aox1, one Aox2a and an Aox2b duplication (named here Aox2b1 and Aox2b2). Expression analyses by semi-quantitative RT-PCR in M. sativa revealed that Aox1, Aox2b1 and Aox2b2 transcripts increased during seed germination. Similar analyses in leaves and roots under different treatments (SA, PEG, H2O2 and cysteine) revealed that these genes are also induced by stress, but with peculiar spatio-temporal differences. Aox1 and Aox2b1 showed basal levels of expression under control conditions and were induced by stress in leaves and roots. Aox2b2 presented a dual behavior, i.e., it was expressed only under stress conditions in leaves, and showed basal expression levels in roots that were induced by stress. Moreover, Aox2a was expressed at higher levels in leaves and during seed germination than in roots and appeared to be not responsive to stress. The Aox expression profiles obtained from a M. truncatula microarray dataset also revealed a stress-induced co-expression of Aox1, Aox2b1 and Aox2b2 in leaves and roots. These results reinforce the stress-inducible co-expression of Aox1/Aox2b in some leguminous plants. Comparative genomic analysis indicates that this regulation is linked to Aox1/Aox2b proximity in the genome as a result of the gene rearrangement that occurred in some leguminous plants during evolution. The differential expression of Aox2b1/2b2 suggests that a second gene has been originated by recent gene duplication with neofunctionalization. © 2013 Elsevier GmbH. All rights reserved.

Sistemas de manejo e efeito residual do potássio na produtividade e nutrição do feijão-caupi

Cowpea [Vigna unguiculata (L) Walp] have great social and economic importance for the Para State. It grows well in areas with low precipitation and two crop cycles can be obtained annually. This study aimed to assess the effect of the residual fertilization from a previous culture (Sorghum bicolor L. Moench) and crop systems on cowpea yield and macronutrient concentration on leaves of three cowpea cultivars (BRSMilênio, BRS-Urubuquara e BRS-Guariba). The study was conducted at the UFRA. The treatments were two crop systems (minimum tillage and conventional), four levels of potassium (50, 100, 200 e 300 kg de KCl ha-1 applied to a previous sorghum culture) and the three cowpea cultivars. Treatments were organized as a three (4 x 2 x 3) factor experiment on a randomized complete block design. The soil was a yellow latosol. In each experiment plot five plants were selected to determine shoot dry matter and foliar nutrient concentration. Grain yield was determined after harvesting all plants on the experiment plot. The residual KCl fertilization affected foliar nutrient content, but did not affect shoot dry mass or yield of grain. Yield was higher in the minimum tillage system. Highest yield (1590 kg ha-1) was recorded int the cv. 'Guariba' when 100 kg of KCl ha-1 had been used in the previous crop. The highest content of leaf N and K was found in cowpea under minimum tillage system. The amount of P and Mg were higher in the conventional system whereas the amount of Ca did not change.