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.

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

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Resistência de genótipos de feijão-vagem ao ataque de Zabrotes subfasciatus (Bohemann, 1833) (Coleoptera: Chrysomelidae)

O feijão-vagem (Phaseolus vulgaris L.) pode sofrer ataque de carunchos, como os da espécie Zabrotes subfasciatus (Bohemann, 1833) (Coleoptera: Chrysomelidae), afetando, diretamente, a qualidade das vagens e sementes e deixando-as inviáveis para o consumo e comercialização. A fim de buscar uma alternativa para o controle deste inseto, foram determinados os tipos e graus de resistência, envolvidos em genótipos de feijão-vagem, ao ataque de Z. subfasciatus, em testes com e sem chance de escolha. Os genótipos utilizados foram UEGD7, UEG05, UEG06, UEG11, UEG13, UEG15, UEG18, UEG19 e UEG26, com quatro repetições. No teste com chance de escolha, os parâmetros avaliados foram o número de ovos viáveis, inviáveis e total e a atratividade dos adultos pelos genótipos. No teste sem chance de escolha, foram avaliados o número de ovos viáveis, inviáveis e total, massa seca consumida e massa seca consumida por inseto, massa e longevidade dos adultos, período de ovo a adulto, razão sexual e número e percentagem de insetos emergidos. Concluiu-se que o genótipo UEG05 foi o menos preferido por Z. subfasciatus, para oviposição, em teste com chance de escolha. O UEG13 foi altamente resistente e UEG05, UEG15 e UEG19 foram moderadamente resistentes. O UEG18 foi susceptível e os demais genótipos foram altamente susceptíveis a Z. subfasciatus.

Resistência de genótipos de feijoeiro a Zabrotes subfasciatus (Bohemann, 1833) (Coleoptera: Bruchidae)

Durante o armazenamento de grãos de feijão (Phaseolus vulgaris L.), o rendimento pode ser reduzido devido às infestações de carunchos como os da espécie Zabrotes subfasciatus (Bohemann, 1833) (Coleoptera: Bruchidae). O ataque desse inseto afeta diretamente a qualidade dos grãos, além de facilitar a entrada de patógenos, tornando-os inviáveis para o consumo e/o comércio. Com a finalidade de buscar uma estratégia alternativa para o controle deste caruncho, avaliou-se a possível resistência de linhagens quase isogênicas contendo arcelina, linhagens selvagens contendo arcelina e cultivares comerciais de feijoeiro, em laboratório (T= 25±2° C, U.R.= 70±10% e fotoperíodo= 12h). Foram utilizados frascos contendo 10 g de grãos dos genótipos, os quais foram infestados por uma semana com sete casais do caruncho. Vinte e um dias após a infestação, os grãos foram avaliados contando-se o número de ovos viáveis. A partir de 25 dias da infestação, os grãos foram observados diariamente avaliando-se o número e o peso dos insetos emergidos, a viabilidade larval, o ciclo biológico (ovo-adulto) e o peso de grãos consumidos. Empregou-se um delineamento inteiramente casualizado, com oito repetições. Os genótipos Arc.2, Arc.3, Arc.4, Arc.3S e Ipa 6 expressaram baixos níveis de não-preferência para oviposição e foram classificados como deterrentes. Os genótipos Arc.1S e Arc.1 expressaram elevados níveis de antibiose; Arc.2, Arc.3 e Arc.4 apresentam o mesmo mecanismo, porém, em níveis inferiores.

Resistance of wild and near isogenic bean lines with arcelin variants to Zabrotes subfasciatus (Boheman): I - Winter crop

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Efeito de genótipos de feijoeiro, óleo de nim em diferentes formulações e período residual no controle de Zabrotes subfasciatus (Boheman, 1833)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Categorias, níveis e causas de resistência de genótipos de feijoeiro a Zabrotes subfasciatus (Boheman, 1833) (Coleoptera: Bruchidae)

Pós-graduação em Agronomia (Entomologia Agrícola) - FCAV

Effects of different formulations of neem oil-based products on control Zabrotes subfasciatus (Boheman, 1833) (Coleoptera: Bruchidae) on beans

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Divergência genética de genótipos de feijoeiro a infestação de Zabrotes subfasciatus (Bohemann) (Coleoptera: Bruchidae)

The aim of this work was to evaluate the genetics divergence of bean genotypes in relation to the oviposition, feeding and development of Zabrotes subfasciatus, determining the degrees of resistance to the weevil. The genotypes used were: IAC Carioca-Tybata, IAC Fortaleza, IAPAR 81, IAC Carioca-Ete, IAC Galante, IAC Harmonia, IAC Una, IAC Diplomata, BRS Supremo and RAZ 49. Tests were performed in laboratory under controlled humidity, temperature and photophase conditions. In free choice test, 10 g of bean genotypes seeds were distributed in circular openings placed equidistant from each other in aluminum trays, where 70 couples were released. The attractiveness was evaluated 24 hours and seven days after the experiment started, and then the number of eggs was evaluated. In non choice test, 10 g of seeds were used where seven couples of Z. subfasciatus, 24 hours-old, were released, remaining seven days, and after the adults retreat, the total number of eggs, viable and unviable eggs, the number and percentage of emerged adults, weight, longevity and period from egg to adult of males and females, sex ratio, dry mass and dry mass consumed by insect were evaluated. In the genotype IAC Harmonia was observed the lower oviposition; RAZ 49 was the most non preference-type resistant for feeding and/or antibiosis-type resistant; BRS Supremo, IAC Carioca-Ete and IAPAR 81 are no preference for feeding and/or antibiosis-type moderate resistant; IAC Galante is susceptible and the other genotypes are highly susceptible to Z. subfasciatus.

Intraspecific competition in Zabrotes subfasciatus: Physiological and behavioral adaptations to different amounts of host

The effects of competition on populations of the bean weevil Zabrotes subfasciatus were analyzed during 41 generations under different competition levels. Three competition environments were established by maintaining the number of couples (6) and varying the amount of available host seeds: HC, high (limited availability of host: 1.35 g); IC, intermediate (intermediate availability of host: 6 g); and LC, low competition (abundance of host: 36 g). It was found that the distribution of the eggs laid on grains was different among treatments: in LC, for example, although females showed high fecundity (35.4 +/- 5.6 eggs/female) the number of eggs laid on each grain was small (1.2 +/- 0.4 eggs on each seed), thus avoiding larval competition of their offspring; whereas in HC treatment, females showed low fecundity (27.04 +/- 4.5 eggs/female) but laid many eggs on each grain (15.03 +/- 4.3 eggs). There were no changes in the ability to respond to different amounts of host via oviposition behavior (egg distribution) during 41 generations. However, HC females had more offspring than LC females under HC conditions. This suggests that HC insects evolved toward higher fitness in crowded conditions. In addition, after inverting the competition level, insects behaved independently of the treatment conditions they experienced through generations, thus showing that oviposition behavior is flexible. Taken together, our results show that Z. subfasciatus presents a broad range of behavioral and physiological responses which allows for quick and reversible adjustments to sudden changes in the amount of resources.

Effect of trypsin inhibitor from Crotalaria pallida seeds on Callosobruchus maculatus (cowpea weevil) and Ceratitis capitata (fruit fly)

GOMES, Carlos E. M. et al. Effect of trypsin inhibitor from Crotalaria pallida seeds on Callosobruchus maculatus (cowpea weevil) and Ceratitis capitata (fruit fly). Plant Physiology and Biochemistry (Paris), v. 43, n. 12, p. 1095-1102, 2005.ISSN 0981-9428. DOI:10.1016/j.plaphy.2005.11.004.

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

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

Resistência de genótipos de feijão-vagem ao ataque de bruquíneos, em condições de laboratório

The aim of this paper was to determine resistance types of snap beans genotypes under infestation of bruchins Zabrotes subfasciatus (Bohemann, 1833) of Acanthoscelides obtectus (Say, 1831) in a no choice and free choice tests. The treatments were arranged in a completely randomized design with four and five replications, respectively. In no choice test, it was evaluated the total number of viable and unviable eggs, dry weight of the consumed food, percentage of emerged insects, weight of insects, longevity of adults and biological cycle of egg to adult. In a free choice test, the number of attracted insects for each genotype and total number of eggs were evaluated. The genotype ARFVI047 presents oviposition non-preference resistance type to Z. subfasciatus. The genotype ARFVI008 presents resistance type of the antibiosis to Z. subfasciatus. The genotypes ARFVI006, ARFVI008 and ARFVI029 present resistance of non-preference for oviposition type to Z. subfasciatus and the genotype HAV 56 black seed presents resistance of non-preference type for feeding of A. obtectus.

Temporal variation in habitat use by nekton in a subtropical estuarine system

Utilisation by fish of different estuarine habitats is known to vary at many different temporal scales, however no study to date has examined how utilisation varies at all the relevant times scales simultaneously. Here, we compare the utilisation by fish of sandy, intertidal foreshore habitats in a subtropical estuary at four temporal scales: between major spawning periods (spring/ summer and winter), among months within spawning periods, between the full and new moon each month, and between night and day within those lunar phases. Comparisons of assemblage composition, abundance of individuals and of fish in seven different,ecological guilds' were used to identify the temporal scales at which fish varied their use of unvegetated sandy habitats in the lower Noosa Estuary, Queensland, Australia. Fish assemblages were sampled with a seine net at three different regions. The most numerically dominant species caught were southern herring (Herklotsichthys castelnaui: Clupeidae), sand whiting (Sillago ciliata: Sillaginidae), weeping toadfish (Torquigener pleurogramma: Tetraodomidae), and silver biddy (Gerres subfasciatus: Gerreidae). Considerable variation at a range of temporal scales from short term (day versus night) to longer term (spawning periods) was detected for all but one of the variables examined. The clearest patterns were observed for diurnal effects, where generally abundance was greater at night than during the day. There were also strong lunar effects, although there were no consistent patterns between full moon and new moon periods. Significant differences among months within spawning periods were more common than differences between the actual spawning periods. The results clearly indicate that utilisation of sandy, unvegetated estuarine habitats is very dynamic and highly variable in space and time. (c) 2006 Elsevier B.V. All rights reserved.