Purificação, caracterização e atividade bioinseticida de um inibidor de tripsina de sementes de Crotalaria pallida

A proteinaceous trypsin inhibitor was purified from Crotalaria pallida seeds by ammonium sulphate fractionation, affinity chromatography on immobilized Trypsin-Sepharose and TCA precipitation. The trypsin inhibitor, named ITC, had Mr of 32.5 kDa by SDS-PAGE and was composed by two subunits with 27.7 and 5.6 kDa linked by disulphide bridges, a typical characteristic of Kunitz-Inhibitor family. ITC was stable until 50°C, and at 100°C its residual activity was of about 60%. Also, ITC was stable at pHs 2 to 12. The inhibition of trypsin by ITC was non-competitive, with a Ki of 8,8 x 10-7M. ITC inhibits weakly other serine proteinases such as chymotrypsin and elastase. The inhibition of papain (44% of inhibition), a cysteine proteinase was an indicative of the bi-functionality of ITC. In vitro assays against digestive proteinases from several Lepdoptera, Diptera and Coleoptera pests were made. ITC inhibited in 100% digestive enzymes of Ceratitis capitata (fruit fly), Spodoptera frugiperda and Alabama argillacea, the last one being a cotton pest. It also inhibited in 74.4% Callosobruchus maculatus (bean weevil) digestive enzymes, a Coleoptera pest. ITC, when added in artificial diet models, affected weakly the development of C. capitata larvae and it had a WD50 of 2.65% to C. maculatus larvae

Purificação, caracterização e análise da atividade bioinseticida, de um inibidor de tripsina em sementes de catanduva (Piptadenia moniliformis)

One Kunitz-type trypsin inhibitors (PmTI) was purified from Piptadenia moniliformis seeds, a tree of the sub-family Mimosoideae, by TCA precipitation, affinity chromatography on immobilized trypsin-Sepharose, DEAE cellulose (ion exchange) and Superose 12 (molecular exclusion) column FPLC/AKTA. The inhibitor has Mr of 25 kDa by SDS-PAGE and chromatography molecular exclusion. The N-terminal sequence of this inhibitor showed high homology with other family Kunitz inhibitors. This also stable variations in temperature and pH and showed a small decrease in its activity when incubated with DDT in the concentration of 100mM for 120 minutes. The inhibition of trypsin by PmTI was competitive, with Ki of 1.57 x10-11 M. The activity of trypsin was effectively inhibited by percentage of inhibition of 100%, among enzymes tested, was not detected inhibition for the bromelain, was weak inhibitor of pancreatic elastase (3.17% of inhibition) and inhibited by 76.42% elastase of neutrophils, and inhibited in a moderate, chymotrypsin and papain with percentage of inhibition of 42.96% and 23.10% respectively. In vitro assays against digestive proteinases from Lepidoptera, Diptera and Coleoptera pests were carried out. Several degrees of inhibition were found. For Anthonomus grandis and Ceratitis capitata the inhibition was 89.93% and 70.52%, respectively, and the enzymes of Zabrotes subfasciatus and Callosobruchus maculatus were inhibited by 5.96% and 9.41%, respectively, and the enzymes of Plodia. interpunctella and Castnia licus were inhibited by 59.94% and 23.67, respectively. In vivo assays, was observed reduction in the development of larvae in 4rd instar of C. capitata, when PmTI was added to the artificial diet, getting WD50 and LD50 of 0.30% and 0.33%, respectively. These results suggest that this inhibitor could be a strong candidate to plant management programs cross transgenic

Microlepidoptera of Illinois hill prairies

Among insects, which are the most diverse eukaryotic group on earth, Lepidoptera is one of four enormously diverse orders, with approximately 10,000 described species in North America. Within the order, Nearctic “microlepidoptera,” which represent an overwhelmingly large percentage of diversity within the order, remain poorly known despite their ecological importance in many plant communities. In this thesis, I undertook several studies of microlepidoptera diversity in a natural community type (hill prairie) and a managed community type (biofuel feedstock). In two Illinois hill prairies differing in size, latitude, and plant composition, alpha diversity of Pyraloidea and Tortricidae was similar, but the prairies were found to support different sets of species of these moth groups. It is concluded that the similarity in alpha diversity occurs because the larger prairie supports primarily a complement of moth species that feed as larvae on prairie plants (especially species of Asteraceae and Fabaceae), whereas the moths collected in the small prairie represent relatively few prairie-associated species, plus a large component of species that feed as larvae on deciduous trees that surround the prairie. This agrees with the finding of high beta diversity of moths between the sites, which reflects a high level of larval hostplant specificity in most species of Pyraloidea and Tortricidae. Based on published information plus observations made on microlepidoptera collected during the course of this study, 31 families of basal microlepidoptera were reviewed with an aim toward evaluating the likelihood of their including species that are dependent on tallgrass prairie. Of these families, 12 were evaluated as possible, and two as likely or certain, to include prairie-dependent species. In a comparison of moth diversity in light-trap samples from corn, miscanthus, switchgrass, and native prairie, alpha diversity was highest in prairie and was higher in switchgrass than in the other two biofuel crops. Moth species complements generally were similar among the biofuel crops, and prairie shared higher species complementarity with switchgrass than with corn or miscanthus. These findings suggest that large-scale conversion of land to biofuel crops may, to a substantial degree, detrimentally affect arthropod biodiversity, with a resulting loss of valuable arthropod-derived ecosystem services both within the cropping systems and in the surrounding landscape. During the course of this study, rearing efforts yielded two species of moths of the family Gelechiidae, both of which are monophagous leaf feeders on leadplant, Amorpha canescens (Fabaceae). Because these moths are restricted to tallgrass prairie, they are likely to be of interest to conservation biologists. In the interest of naming the moths to facilitate communication regarding them, and to augment our taxonomic knowledge of their respective genera, the moths are described, and diagnoses are provided to differentiate them from similar, related species.

Oligomerization of Cry9Aa in solution without receptor binding, is not related with insecticidal activity

Background: Bacillus thuringiensis Cry toxins bind with different insect midgut proteins leading to toxin oligomerization, membrane insertion and pore formation. However, different Cry toxins had been shown to readily form high molecular weight oligomers or aggregates in solution in the absence of receptor interaction. The role of Cry oligomers formed in solution remains uncertain. The Cry9A proteins show high toxicity against different Lepidoptera, and no-cross resistance with Cry1A. Results: Cry9Aa655 protein formed oligomers easily in solution mediated by disulfide bonds, according to SDS-PAGE analysis under non-reducing and reducing conditions. However, oligomerization is not observed if Cry9Aa655 is activated with trypsin, suggesting that cysteine residues, C14 and C16, located in the N-terminal end that is processed during activation participate in this oligomerization. To determine the role of these residues on oligomerization and in toxicity single and double alanine substitution were constructed. In contrast to single C14A and C16A mutants, the double C14A–C16A mutant did not form oligomers in solution. Toxicity assays against Plutella xylostella showed that the C14A–C16A mutant had a similar insecticidal activity as the Cry9Aa655 protein indicating the oligomers of Cry9Aa formed in solution in the absence of receptor binding are not related with toxicity. Conclusions: The aggregation of Cry9Aa655 polypeptides was mediated by disulfide bonds. Cry9Aa655 C14 and C16C are involved in oligomerization in solution. These aggregate forms are not related to the mode of action of Cry9Aa leading to toxicity.

Biologia da traca do tomateiro sob condicoes de laboratorio.

Estudaram-se, no CPATSA-EMBRAPA em Petrolina, PE, sob condicoes de laboratorio, os aspectos biologicos de Scrobipalpula absoluta (Meyrick 1917) (Lepidoptera,Gelechiidae), importante praga do tomateiro, Iniciou-se a criacao com larvas coletadas no campo, em cultura de tomate, acondicionadas, em placas de Petri, sobre papel de filtro e folhas de tomateiro, variedade IPA-2. Os adultos foram mantidos em frascos de vidro revestidos com papel sulfite, tampados com tecido branco de nailon preso com elastico e alimentados, com uma solucao de sacarose a 10%, embebida em algodao. Obtiveram-se os seguintes dados: periodo de pre-ovoposicao 2,35 dias; numero de ovos por femea 55,16; viabilidade de ovos 44,46%, periodo de incubacao 4,30 dias; periodo de oviposicao 3,77 dias; periodo larval 10,95 dias com quatro instares; viabilidade larval 20,97%; periodo pupal 6,15 dias; viabilidade pupal 68,19%; longevidade de adultos machos e femeas 9,69 e 11,52 dias, respectivamente; ciclo biologico (ovo-morte do adulto) 38,12 dias.

Insetos-praga da macieira: reconhecendo os inimigos.

2016

Natural parasitism of lepidopteran eggs by Trichogramma species (Hymenoptera: Trichogrammatidae) in agricultural crops in Minas Gerais, Brazil.

The genus Trichogramma Westwood (Hymenoptera: Trichogrammatidae) includes insect egg parasitoids that are widely used throughout the world as control agents of pest insects. The aim of this study was to identify the species of Trichogramma naturally associated with the eggs of lepidopteran pests of the following agricultural and horticultural crops: collards, Brassica oleracea L. (Brassicales: Brassicaceae); papaya, Carica papaya L. (Capparales: Caricaceae); tomato, Lycopersicon esculentum Mill. (Solanales: Solanaceae); cassava, Manihot esculenta Crantz (Malpighiales: Euphorbiaceae); banana, Musa sp. L. (Zingiberales: Musaceae); passion fruit, Passiflora sp. Degener (Malpighiales: Passifloraceae); sugarcane, Saccharum sp. L. (Poales: Poaceae); and corn (maize), Zea mays L. (Poales: Poaceae); and an invasive species (Sodom?s apple milkweed, Calotropis procera Aiton; Gentianales: Apocynaceae) in the semiarid region of Minas Gerais, Brazil. We report natural parasitism by Trichogramma in eggs of Agraulis vanillae vanillae (L.) (Lepidoptera: Nymphalidae), Antichloris eriphia F. (Lepidoptera: Arctiidae), Danaus sp. (L.) (Lepidoptera: Nymphalidae), Diatraea saccharalis F. (Lepidoptera: Crambidae), Erinnyis ello L. (Lepidopera: Sphingidae), and Protambulyx strigilis L. (Lepidopera: Sphingidae). In total, 2,242 specimens of Trichogramma were obtained, belonging to the species T. pretiosum Riley, T. manicobai Brun, Moraes & Soares, T. marandobai Brun, Moraes & Soares, and T. galloi Zucchi. These species of Trichogramma may be candidates for biological control programs of lepidopteran pests in the semiarid region of Minas Gerais and in other semiarid regions.

Biological activity of astilbin from Dimorphandra mollis against Anticarsia gemmatalis and Spodoptera frugiperda

Astilbin was isolated in high yield from Dimorphandra mollis, and its insecticidal and growth inhibiting activity by stomach ingestion were evaluated against Anticarsia gemmatalis and Spodoptera frugiperda. The insecticidal activity of astilbin, the weight reduction of the larval phase and the prolongation of the larval and pupal phases were verified for both species. Astilbin was identified on the base of its NMR, MS and physical data. (C) 2002 Society of Chemical Industry.

Intención de emprendimiento y liderazgo en Colombia

La relación entre el liderazgo y emprendimiento es un tema de estudio y análisis relativamente nuevo. El término de liderazgo ha sido estudiado y enfocado a otras áreas de estudio, pero en cuanto a la relación existente con emprendimiento se presentan aún carencias conceptuales. A lo largo del presente estudio, se hace una revisión de la literatura la cual permite identificar y analizar estos conceptos y su relación en la última década en Colombia, consultando literatura comprendida en bases de datos en línea tales como, Redalyc, Scielo y Dialnet. En este documento, se destacan los atributos de un emprendedor y de un líder, igualmente se recogen los modelos de emprendimiento, las teorías de liderazgo, y los elementos que caracterizan cada concepto. Finalmente, se evidencia una dicotomía entre el líder y el emprendedor y se concluye que el liderazgo es una característica intrínseca del emprendimiento; sin embargo, el liderazgo no es por regla utilizado para emprender.

Artrópodos-fitófagos em frutíferas nativas e pequenas frutas.

Neste trabalho são apresentadas informações sobre as principais espécies pragas associadas à amoreira-preta, mirtileiro, pitangueira e araçazeiro, sendo dado destaque ao desenvolvimento da mosca-das-frutas sul-americana (Anastrepha fraterculus) nestes hospedeiros, bem como ao estádio fenológico dos frutos susceptível a praga.

Ocorrência de espécies de Helicoverpa em cultivos de tomateiro no Planalto Central Brasileiro.

Esse estudo teve como objetivo determinar quais espécies de Helicoverpa estão atualmente associadas ao broqueamento de tomate na região do Planalto Central Brasileiro.

Wavelet analysis and hierarchical clustering of ofm phenotypic population's variations originated from southern america and europe and attempts of a first relation to molecular divergence.

In this work we compare Grapholita molesta Busck (Lepidoptera: Tortricidae) populations originated from Brazil, Chile, Spain, Italy and Greece using power spectral density and phylogenetic analysis to detect any similarities between the population macro- and the molecular micro-level. Log-transformed population data were normalized and AR(p) models were developed to generate for each case population time series of equal lengths. The time-frequency/scale properties of the population data were further analyzed using wavelet analysis to detect any population dynamics frequency changes and cluster the populations. Based on the power spectral of each population time series and the hierarchical clustering schemes, populations originated from Southern America (Brazil and Chile) exhibit similar rhythmic properties and are both closer related with populations originated from Greece. Populations from Spain and especially Italy, have higher distance by terms of periodic changes on their population dynamics. Moreover, the members within the same cluster share similar spectral information, therefore they are supposed to participate in the same temporally regulated population process. On the contrary, the phylogenetic approach revealed a less structured pattern that bears indications of panmixia, as the two clusters contain individuals from both Europe and South America. This preliminary outcome will be further assessed by incorporating more individuals and likely employed a second molecular marker.

Eficiência de tratamentos inseticidas no controle da traça-do-cacho da videira.

2015