Vicilins (7S storage globulins) of cowpea (Vigna unguiculata) seeds bind to chitinous structures of the midgut of Callosobruchus maculatus (Coleoptera: Bruchidae) larvae

The presence of chitin in midgut structures of Callosobruchus maculatus larvae was shown by chemical and immunocytochemical methods. Detection by Western blotting of cowpea (Vigna unguiculata) seed vicilins (7S storage proteins) bound to these structures suggested that C. maculatus-susceptible vicilins presented less staining when compared to C. maculatus-resistant vicilins. Storage proteins present in the microvilli in the larval midgut of the bruchid were recognized by immunolabeling of vicilins in the appropriate sections with immunogold conjugates. These labeling sites coincided with the sites labeled by an anti-chitin antibody. These results, taken together with those previously published showing that the lower rates of hydrolysis of variant vicilins from C. maculatus-resistant seeds by the insect's midgut proteinases and those showing that vicilins bind to chitin matrices, may explain the detrimental effects of variant vicilins on the development of C. maculatus larvae.

The resistance of seeds of cowpea (Vigna unguiculata) to the cowpea weevil (Callosobruchus maculatus)

Cowpea (Vigna unguiculata) seeds are heavily damaged during storage by the bruchid Callosobruchus maculatus. Seeds of some Nigerian varieties showed a strong resistance to this bruchid. By utilizing biochemical and entomological techniques we were able to rule out the paticipation of proteolytic enzyme (trypsin, chimotrypsin, subtilisin and papain) inhibitors, lectins, and tannins in the resistance mechanisms. Fractionation of the seed meal of a resistant variety suggests that the factor(s) responsible for the effect is (are) concentrate in the globulin fraction.

Characterization of cowpea genotype resistance to Callosobruchus maculatus

The objective of this work was to characterize the resistance of 50 cowpea (Vigna unguiculata) genotypes to Callosobruchus maculatus. A completely randomized design with five replicates per treatment (genotype) was used. No-choice tests were performed using the 50 cowpea genotypes to evaluate the preference for oviposition and the development of the weevil. The genotypes IT85 F-2687, MN05-841 B-49, MNC99-508-1, MNC99-510-8, TVu 1593, Canapuzinho-1-2, and Sanzi Sambili show non-preference-type resistance (oviposition and feeding). IT81 D-1045 Ereto and IT81 D-1045 Enramador exhibit antibiosis against C. maculatus and descend from resistant genitors, which grants them potential to be used in future crossings to obtain cowpea varieties with higher levels of resistance.

Lima bean (Phaseolus lunatus) seed coat phaseolin is detrimental to the cowpea weevil (Callosobruchus maculatus)

The presence of phaseolin (a vicilin-like 7S storage globulin) peptides in the seed coat of the legume Phaseolus lunatus L. (lima bean) was demonstrated by N-terminal amino acid sequencing. Utilizing an artificial seed system assay we showed that phaseolin, isolated from both cotyledon and testa tissues of P. lunatus, is detrimental to the nonhost bruchid Callosobruchus maculatus (F) (cowpea weevil) with ED50 of 1.7 and 3.5%, respectively. The level of phaseolin in the seed coat (16.7%) was found to be sufficient to deter larval development of this bruchid. The expression of a C. maculatus-detrimental protein in the testa of nonhost seeds suggests that the protein may have played a significant role in the evolutionary adaptation of bruchids to legume seeds.

Purification, partial characterization and role in lipid transport to developing oocytes of a novel lipophorin from the cowpea weevil, Callosobruchus maculatus

Lipid transport in arthropods is achieved by highly specialized lipoproteins, which resemble those described in vertebrate blood. Here we describe purification and characterization of the lipid-apolipoprotein complex, lipophorin (Lp), from adults and larvae of the cowpea weevil Callosobruchus maculatus. We also describe the Lp-mediated lipid transfer to developing oocytes. Lps were isolated from homogenates of C. maculatus larvae and adults by potassio bromide gradient and characterized with respect to physicochemical properties and lipid content. The weevil Lp (465 kDa) and larval Lp (585 kDa), with hydrated densities of 1.22 and 1.14 g/mL, contained 34 and 56% lipids and 9 and 7% carbohydrates, respectively. In both Lps, mannose was the predominant monosaccharide detected by paper chromatography. SDS-PAGE revealed two apolipoproteins in each Lp with molecular masses of 225 kDa (apolipoprotein-I) and 79 kDa (apolipoprotein-II). The lipids were extracted and analyzed by thin-layer chromatography. The major phospholipids found were phosphatidylserine, phosphatidylcholine and phosphatidylethanolamine in adult Lp, and phosphatidylcholine, phosphatidylethanolamine and sphingomyelin in larval Lp. Hydrocarbons, fatty acids and triacylglycerol were the major neutral lipids found in both Lps. Lps labeled in the protein moiety with radioactive iodine (125I-iodine) or in the lipid moiety with fluorescent lipids revealed direct evidence of endocytic uptake of Lps in live oocytes of C. maculatus.

Identification of Albizia lebbeck seed coat chitin-binding vicilins (7S globulins) with high toxicity to the larvae of the bruchid Callosobruchus maculatus

Seed coat is a specialized maternal tissue that interfaces the embryo and the external environment during embryogenesis, dormancy and germination. In addition, it is the first defensive barrier against penetration by pathogens and herbivores. Here we show that Albizia lebbeck seed coat dramatically compromises the oviposition, eclosion and development of the bruchid Callosobruchus maculatus. Dietary supplementation of bruchid larvae with A. lebbeck seed coat flour causes severe weight loss and reduces survival. By means of protein purification, mass spectrometry and bioinformatic analyses, we show that chitin-binding vicilins are the main source of A. lebbeck tegumental toxicity to C. maculatus. At concentrations as low as 0.1%, A. lebbeck vicilins reduce larval mass from 8.1 ± 1.7 (mass of control larvae) to 1.8 ± 0.5 mg, which corresponds to a decrease of 78%. Seed coat toxicity constitutes an efficient defense mechanism, hindering insect predation and preventing embryo damage. We hypothesize that A. lebbeck vicilins are good candidates for the genetic transformation of crop legumes to enhance resistance to bruchid predation.

Resistência de genótipos de caupi ao caruncho

A utilização de resistência genética ao ataque de Callosobruchus maculatus (Fabr.) tem sido alvo de investigação científica, especialmente no que diz respeito à identificação de fontes de resistência. O presente trabalho objetivou incorporar, ao grupo de caracteres desejáveis para o cultivo de caupi (Vigna unguiculata (L.) Walp.), resistência genética ao caruncho (C. maculatus). Foram realizadas hibridações dos genótipos IT81D-1045 e IT81D-1064 (portadores de resistência ao inseto) com CNCx 252-1E/FB, CNCx 187-22D-1 e BR 1-Poty (capazes de transferir resistência a viroses, tolerância à seca, formação de grãos com padrão comercial, elevado potencial de produção e adaptabilidade a diferentes condições ambientais). Populações segregantes obtidas destes cruzamentos foram conduzidas pelo método SPD (descendência de uma única vagem), e na geração F5 foram realizadas seleções individuais. As linhagens obtidas foram avaliadas em conjunto com materiais de origens diferentes, utilizando-se parâmetros associados à infestação da praga. Foi constatado que os genótipos avaliados apresentaram variabilidade quanto à preferência à postura, número de insetos emergidos e número de sementes danificadas; as linhas EVx 37-15E e EVx 37-2E foram as que sofreram menor dano causado pelo caruncho; as variáveis número de ovos, número de insetos emergidos e número de sementes danificadas mostraram-se positiva e significativamente correlacionadas entre si; o grupo das linhagens que descendem de genitores resistentes apresenta valores significativamente inferiores aos obtidos pelas demais, o que indica que a resistência ao inseto se transmite geneticamente.

Insecticidal and antifungic proteins of the latex from Manihot glaziovii Muell. Arg.

Analysis of the latex from Manihot glaziovii showed the presence of various enzymatic and inhibitory activities. The latex also presented an inhibitory effect on the development of cowpea weevil (Callosobruchus maculatus) in an artificial seed system and on the development, in an in vitro assay, of phytopathogenic fungi Colletotrichum gloesporioides, Fusarium solani and Macrophomina phaseolina. These results suggest the presence of substances, some of them of protein nature, involved in plant defense mechanisms in this exudation product.

Variation of some chemical and functional properties of Bambara groundnut (Voandzeia Subterranean L. Thouars) during sort time storage

Abstract The storage susceptibility of Bambara groundnut (B. G.) (Voandzeia Subterranean (L.) Thouars) to Callosobruchus maculatus and chemical and functional properties of 11 varieties form Far-North of Cameroon were investigate using standard analytical methods. Storage susceptibility shown that, after five months within treatment, C. maculatus destroy 10 to 50% of grains. The chemical characteristics of none attack grains of 11 varieties were range to 18.64 at 21.08%, 6.85 at 7.44%, 49.75 at 52.68% and to 6.05 at 7.55% respectively for protein, fat, starch and free carbohydrate. These chemical characteristics significantly (p < 0.05) decreases form attacks varieties. For the functional parameters, the none attacks grains was range of 130 at 135%, 19.15 at 20.91%, 18.20 at 21.13%, 2.76 at 3.21% and of 8.54 at 10.14% respectively for water capacity absorption, solubility index, gel length, ash and humidity. The results of this study indicated that storage susceptibility, chemical and functional properties of B. G. be dependant to the varieties.

Estolomimus (Coleoptera, Cerambycidae, Desmiphorini): novas espécies e chave

New species described from Brazil: Estolomimus maculatus (Minas Gerais and São Paulo), E. transversus (Espírito Santo), E. lichenophorus (São Paulo), and from Bolivia: E. abjunctus (Beni). A key to the species is added.

Novos táxons neotropicais de Desmiphorini (Coleoptera, Cerambycidae, Lamiinae)

New species described from Brazil: Curuapira tuberosa (Espírito Santo); Stereomerus maculatus (Bahia); Gyrpanetes oryba (Espírito Santo); Desmiphora (D.) santossilvai (Pará); Desmiphora (D.) uniformis (Bahia). Icublabia gen. nov., type-species, I. multispinosa sp. nov. from Ecuador (Napo) are described. Curuapira exotica Martins & Galileo, 1998, originally described from Venezuela, is recorded from Brazil (Amazonas, Rondônia and Mato Grosso).

Eryphus Perty, 1832 e Tacyba, um novo gênero de Heteropsini (Coleoptera, Cerambycidae, Cerambycinae)

Eryphus Perty, 1832 and Tacyba, a new genus of Heteropsini (Coleoptera, Cerambycidae). Some species, up to now, included in Callideriphus Blanchard, 1851 are rearranged in: a) those congeneric with Callideriphus grossipes Blanchard, 1851 and b) not congeneric. The first set of species will be treated in a future paper; the second one, on the other hand, is subdivided into Eryphus Perty, 1832 and Tacyba gen. nov. Eryphus Perty, 1832 (type species: Eryphus bipunctatus Perty, 1832), a valid genus, is redescribed and a key for the species is also provided. The following species are transferred to Eryphus: E. bivittatus (Melzer, 1934) comb. nov., E. carinatus (Zajciw, 1970) comb. nov., E. flavicollis (Fisher, 1938) comb. nov., E. laetus (Blanchard, 1851) comb. nov., E. marginatus (Zajciw, 1970) comb. nov., E. picticollis (Gounelle, 1911) comb. nov., E. transversalis (Fairmaire & Germain, 1864) comb. nov. New synonym proposed: Eryphus bipunctatus Perty, 1832 = Callideriphus atricollis Melzer, 1931. New taxa described: Eryphus tacuarembo sp. nov. (Uruguay, Tacuarembó), E. carioca sp. nov. (Brazil, Rio de Janeiro); Tacyba gen. nov. (type species: Callideriphus maculatus Cerda, 1988). Species transferred to Tacyba and synonyms: T. maculata (Cerda, 1988) comb. nov., T. tenuis (Blanchard, 1851) comb. nov. = Callideriphus testaceicornis Fairmaire & Germain, 1859 syn. nov. = Callideriphus clathratus Fairmaire & Germain, 1860 syn. nov. = Callideriphus niger Philippi & Philippi, 1864 syn. nov. Callideriphus flavicollis m. quadripunctatus Fuchs, 1961 and Callideriphus flavicollis m. reductus Fuchs, 1961, both names of infrasubspecific category (not available under the rules of ICZN), are herein treated as intraspecific variation of Eryphus picticollis (Gounelle, 1911) which occur in southern Brazil and Argentina.

Determinação dos tipos de resistência em genótipos de feijoeiro ao ataque de Zabrotes subfasciatus (Coleoptera: Bruchidae)

O Zabrotes subfasciatus (Boheman, 1833) (Coleoptera: Bruchidae) é considerado a principal praga do feijão armazenado. O método de controle mais utilizado tem sido o uso de inseticidas; no entanto outros métodos podem ser empregados, como, por exemplo, a resistência de plantas. Este trabalho teve por objetivo determinar os tipos de resistência em 12 genótipos de feijoeiro ao ataque dessa praga. O experimento foi conduzido no Laboratório de Resistência de Plantas a Insetos, do Departamento de Fitossanidade da FCAV\UNESP, Jaboticabal - SP, no período de março a novembro de 2009. Foram realizados testes de não preferência para oviposição, com e sem chance de escolha, e de alimentação, sem chance de escolha. Concluiu-se que os genótipos Dor 391, Dor 476, IAPAR-MD 806 e Raz 59 apresentam resistência do tipo não preferência para oviposição por Z. subfasciatus, em teste com chance de escolha; no teste sem chance de escolha todos os genótipos foram igualmente ovipositados pelo inseto; no teste com chance de escolha, machos e fêmeas foram igualmente atraídos por todos os genótipos testados; e Raz 56, Arc 2, Raz 55, Raz 49 e Raz 59 apresentaram resistência dos tipos não preferência para alimentação e,ou, antibiose.

Estimate of Physiological Age in Females of PASSALUS (Insecta, Coleoptera)

Studies on development of female reproductive system insects have shown its utility in physiological age grading. The irreversible charocter of ovarian alterations correlated with mandibular wear and presence of yellow body, were applied to estimate the physiological age of Passalus convexus and Passalus latifrons females, giving results comparable to other studies and indicating the value of these methods in determination of physiological age.

Contribuição ao conhecimento da bionomia de Passalus convexus Dalman, 1817 e Passalus latifrons Percheron, 1841 (Coleoptera: Passalidae)

Coleópteros da família Passalidae, com distribuição pantropical, parecem contribuir, pelo menos em parte, para a reciclagem dos nutrientes nas florestas, facilitando a penetração de umidade e de microorganismos decompositores no interior dos troncos mortos, pela abertuda de galerias. Aspectos da bionomia de Passalus convexus Dalm. e Passalus latifrons Perch. Foram estudados durante um ciclo sazonal completo, com levantament dos tipos de troncos atacadio, fauna acompanhante e os estágios de decomposição mais propícios à colonização.