Females of the weevil Zabrotes subfasciatus manipulate the size and number of eggs according to the host seed availability

In some insects, the finding of oviposition substrate triggers the uptake into oocytes of yolk proteins that are stored in the fat body during post-embryonic development. The main host of the bean weevil Zabrotes subfasciatus (Coleoptera; Chrysomelidae; Bruchinae; Amblycerini), in which larval resources are the sole source for future egg maturation, is Phaseolus vulgaris. Despite not feeding as adults, females of this species are able to lay eggs after encountering host seeds but it is not known how females react to changes in the availability of bean seeds. In the present study, the behaviour of Z. subfasciatus facing two very different environments for oviposition is investigated, as well as how this influences offspring fitness. The results obtained show that females of Z. subfasciatus react to variations in the availability of seeds belonging to the same host species by adjusting egg size and number. Females on low bean seed density lay larger and fewer eggs than those on high bean seed density, demonstrating a trade-off between these reproductive traits. Moreover, females can adjust egg size to changing levels of host availability during the first 4 days of their oviposition period. Although no difference in offspring weight is found, those from small eggs (low competition environment) result in larger adults. No response to selection on these traits after rearing beetles on the same host for 40 generations is observed. This unresponsiveness may indicate that beetle populations behave according to their reaction norm that already allows rapid adaptation to a varying amount of host-seed availability and better exploitation of the environments of this widespread stored-seed pest.

Host preference of the bean weevil Zabrotes subfasciatus

It is largely known that the range of an insect diet is mostly determined by oviposition behavior, mainly in species with endophytic larvae such as Zabrotes subfasciatus. However, the proximate factors determining host choice and the subsequent steps leading to the expansion or reduction of the host number and occasional host shifts are largely unknown. We analyzed various factors determining host preference of Z. subfasciatus through the evaluation of: (i) oviposition preference of a wild population of Z subfasciatus on the usual host (bean) and unusual hosts (lentil, chickpea and soy), and the performance of the offspring; (ii) artificial selection for increasing preference for hosts initially less frequently chosen; (iii) comparison of oviposition behavior between two different populations (reared for similar to 30 generations in beans or chickpeas, respectively); (iv) oviposition timing on usual and unusual hosts; and (v) identification of preference hierarchies. We found that when using unusual hosts, there is no correlation between performance and preference and that the preference hierarchy changes only slightly when the population passes through several generations on the less frequently accepted host. We also found a positive response to artificial selection for increasing oviposition on the less preferred host; however, when the host-choice experiment involved two varieties of the usual host, the response was faster than when the choice involved usual and unusual hosts. Finally, beetles reared on an unusual host (chickpea) for 26 generations showed similar good fitness on both usual and unusual hosts, indicating that the use of a new host does not necessarily result in the loss of performance on the original host. Nevertheless, this population showed lower fitness on the usual host than that of the original population, suggesting an underlying partial trade-off phenomenon which may contribute to a broadening of diet of this insect species.

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.

Toxicity and repellency of essential oils to Zabrotes subfasciatus (Boheman) (Coleoptera, Chrysomelidae, Bruchinae) in Phaseolus vulgaris L

The effects of tangerine (Phaseolus vulgaris Blanco), lemon (Citrus medica limonum Lush), pear orange (Citrus sinensis L. Osbeck), red copaiba (Copaifera langsdorffii Desf.), rosemary (Baccharis dracunculifolia De Candole), Eucalyptus (Eucalyptus globulus Labillardière and E. citriodora Hook), lemongrass (Cymbopogon citratus Stapf.) and citronella (Cimbopogon nardus Linnaeus) oils at several concentrations on Zabrotes subfasciatus (Boheman) were studied. In toxicity tests, grains of Phaseolus vulgaris L. cv. Rajadinho were impregnated with oils and infested with adults of Z. subfasciatus up to 24 hours old. All tested oils were effective in reducing the viable egg-laying and adult emergence of this pest, in function of the concentrations used, highlighting E. citriodora and E. globulus oils which caused 100% effectiveness from 0.5 mL Kg-1 concentration. In repellency tests, two arenas consisting of plastic containers, connected symmetrically to a central box by two plastic tubes were used. In one of the boxes, untreated beans were placed and on the other ones beans treated with each oil concentration were used. In the central box, five couples of Z. subfasciatus were released. Grains of P. vulgaris treated with oils of E. citriodora, C. citratus and C. oleifera reduced the attraction percentage of Z. subfasciatus adults, while the E. globulus increased this percentage. The percentages of reduced viable eggs ranged from 17.9% (C. medica limonum) to 93.3% (C. nardus), while the reduction on the number of emerged insects was 23.9% and 95.9%, respectively for these same oils.

Oviposition behavior of Zabrotes subfasciatus females (Coleoptera, Bruchidae) under conditions of host deprivation

Oviposition of Zabrotes subfasciatus (Boheman, 1833) on Phaseolus vulgaris (Linnaeus, 1753) was studied immediately after emergence of the adults throughout the females life and in situations of host deprivation lasting for 1 to 10 days. The number of eggs laid daily, longevity, duration of oviposition and distribution of eggs per grain were studied. The number of eggs laid per day varied significantly, with the oviposition peak in the presence of the host (control group) occurring between day 2 and day 5 of oviposition. In the absence of the host, a shift in the oviposition peak to the first day after deprivation was observed, except for the group deprived for one day which showed a peak between days 1 and 4 after introduction of the host. The distribution of the eggs per grain in the control group and in the groups deprived of the host for 2, 5, 8 and 10 days, a larger egg aggregation was observed for all deprived groups compared to the control group.

Oviposition behavior of wild Zabrotes subfasciatus (Coleoptera, Chrysomelidae) females deprived of the host Phaseolus vulgaris (Fabaceae)

The insects oviposition behavior is fundamental to study population dynamics, life history evolution, insect-plant and parasitoid-host interactions. Zabrotes subfasciatus (Boheman, 1833) females oviposition behavior in the presence and absence of a host is unknown. The main objective of this study was to describe in detail the oviposition behavior of host deprived or non-deprived females, and observe how the several situations of deprivation (days without host) influence oviposition. Six groups were assembled, three deprived of the host (for 2, 5 and 8 days) and three control groups (with host), each containing one newly-emerged couple (0-24h) of wild Z. subfasciatus, The non-deprived (control) groups received the hosts every day (5 bean seeds Phaseolus vulgaris (Fabaceae)) and the others were deprived for 2, 5 and 8 days, respectively. For each group 12 repetitions were made. Consequently, 12 couples were host deprived during two days, 12 couples were host deprived during five days and 12 couples were host deprived during eight days. When the seeds of the deprived groups were added the experiments started. There was a control group for each deprived group. The experiments and the insects were maintained at constant temperature 29 ± 2ºC and 70-80% relative humidity. At 15 minutes interval, the number of times the females manifested the different categories of behavior was observed (frequency). The behavior categories were: rest inside the box, locomotion, resource exploration (seeds), copulation and oviposition. The deprived females stayed most of the time in contact with the host to carry out oviposition, while the non-deprived (control) females spent most of the time at rest. This was observed in all the deprivation times. The results show that host deprivation influences the oviposition behavior of the studied species and also shows the flexibility in the oviposition strategies that these females present when the environment changes (absence and presence of resources)

Aspectos biológicos de Zabrotes subfasciatus (Bohemann, 1833) (Coleoptera, Bruchidae) em Phaseolus vulgaris L., cv. Carioca (Fabaceae), sob condições de laboratório

Zabrotes subfasciatus is a serious pest of common beans, P. vulgaris L.. In Brazil there are several studies dealing with resistance of bean genotypes to this insect, while other studies have emphasized the utilization of oils and powders from plants to repel their attack. In this paper, fecundity, fertility, pattern of oviposition, life cycle and longevity were evaluated for a Brazilian stock from the Goiás State on P. vulgaris cv. Carioca, at 30ºC and 70% R.H. The mean fecundity was 38 eggs per female and 73% of viability. Egg laying showed an aggregated pattern. Males and females lived an average of 13 and 9 days, respectively. The total life cycle lasted for about 28 days.

Effect of arcelin protein on the biology of Zabrotes subfasciatus (Boheman 1833), in dry beans

Arcelin is a seed protein found in wild beans (Phaseolus vulgaris) which gives resistance to Mexican bean weevil, Zabrotes subfasciatus (Boheman 1833) (Coleoptera: Bruchidae). Studies were carried out with the objective of estimating the effect of four alleles of protein arcelin (Arc1, Arc2, Arc3 and Arc4) on the biology of Z. subfasciatus. The experiment was carried out in laboratory at Embrapa-Centro Nacional de Pesquisa de Arroz e Feijão, in Santo Antônio de Goiás, GO, Brazil, under non controlled conditions. The highest levels of antibiosis to Z. subfasciatus were observed in Arc1, with reduction in the number of eggs, number of emerged adults, adults longevity. In the line Arc2 only reduction in the number of emerged adults was observed. The lines Arc3 and Arc4 showed low efficiency on the reduction of progeny of Z. subfasciatus and effects in the longevity and egg-adult cycle were not detected. Insect sexual ratio was not altered by the presence of Arc1, Arc2, Arc3 and Arc4 in the seeds.

Estabilidade da resistência a Zabrotes subfasciatus conferida pela proteína arcelina, em feijoeiro

A arcelina é uma proteína encontrada em feijões silvestres e confere resistência ao caruncho-do-feijão, Zabrotes subfasciatus (Boh., 1833) (Coleoptera: Bruchidae). Comparativamente às cultivares suscetíveis Porrillo 70 e Goiano Precoce, estudou-se a estabilidade da resistência, conferida por dois alelos da proteína arcelina, nas linhagens do feijoeiro Arc1 e Arc4, após a criação do inseto por quatro gerações sucessivas no mesmo genótipo. O experimento foi conduzido no Laboratório da Embrapa-Centro Nacional de Pesquisa de Arroz e Feijão (CNPAF), no município de Santo Antônio de Goiás, GO, em condições não-controladas, utilizando-se delineamento experimental inteiramente ao acaso. Observaram-se, nas linhagens portadoras de arcelina, redução do número de ovos e de adultos emergidos, redução do peso dos insetos, e prolongamento do período ovo-adulto. Na linhagem Arc1, a resistência ao Z. subfasciatus foi mais expressiva. Nessa linhagem, os parâmetros número de ovos, número de adultos emergidos, período ovo-adulto, razão sexual e peso de machos não foram alterados, porém observou-se aumento progressivo de peso das fêmeas nas gerações. Na linhagem Arc4, houve redução do período ovo-adulto, embora o número de ovos e de adultos emergidos, razão sexual e peso de machos e fêmeas não tenham sido alterados.

Controle do caruncho-do-feijoeiro Zabrotes subfasciatus com óleos vegetais, munha, materiais inertes e malathion

Na busca de alternativas ao controle químico do caruncho Zabrotes subfasciatus (Coleoptera:Bruchidae), avaliou-se, em feijoeiro (Phaseolus vulgaris L.), o efeito de três genótipos, cv. Carioca e duas linhagens contendo a proteína arcelina (Arc1 e Arc3), e da adição nos grãos armazenados de óleo de soja (Glycine max Merrill); óleo de nim (Azadirachta indica A. Juss.), munha (resíduo de trilha da colheita), calcário dolomítico e terra de formigueiro, comparativamente aos grãos não tratados e ao controle químico com malathion 500 CE. O experimento foi realizado no laboratório da Embrapa-Centro Nacional de Pesquisa de Arroz e Feijão, localizado no Município de Santo Antônio de Goiás, GO, em condições não controladas. Na linhagem Arc1, constatou-se maior proteção aos danos do caruncho, observando-se redução do número de ovos e de adultos emergidos e da porcentagem de sementes danificadas em relação à Arc3 e à cv. Carioca. Quando os grãos foram tratados com malathion, óleo de nim e óleo de soja, observou-se menor número de ovos e de adultos emergidos e redução de danos, e não houve diferença na porcentagem de sementes danificadas entre o malathion e o óleo de nim. A mistura das sementes com terra de formigueiro conferiu baixa proteção ao caruncho, enquanto os tratamentos com munha e calcário dolomítico não apresentaram eficiência na redução da progênie e dos danos de Z. subfasciatus.

Diversidade genética estimada com marcadores ISSR em populações brasileiras de Zabrotes subfasciatus

O objetivo deste trabalho foi estimar a diversidade genética de populações de Zabrotes subfasciatus, por meio de marcadores moleculares ISSR. Foram avaliadas 12 populações, provenientes de oito estados brasileiros, no total de 269 indivíduos. Cinco iniciadores ISSR permitiram a obtenção do total de 51 fragmentos polimórficos. A percentagem média de locos polimórficos, dentro de cada população, foi de 83,8%. A heterozigosidade corrigida de Nei esperada variou de 0,23 a 0,33, com média de 0,29, e o índice de diversidade genética de Shannon e Weaver variou de 0,29 a 0,48, com média de 0,42. No nível de espécie, estes dois índices apresentaram valores de 0,36 e 0,54, respectivamente. A análise de variância molecular mostrou que 66% da variância molecular total pode ser atribuída a diferenças intrapopulacionais e que os 34% restantes podem ser atribuídos a diferenças interpopulacionais. O teste de Mantel mostrou baixa correlação entre: distância geográfica e diferenciação genética, identidade genética e diferenciação genética e, distância genética de Nei e diferenciação genética. As populações brasileiras de Z. subfasciatus possuem baixa diferenciação genética e fraca estruturação geográfica.

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.