Suitability of Arabidopsis thaliana as a model for host plant-Plutella xylostella-Cotesia plutellae interactions

Successful pest management is often hindered by the inherent complexity of the interactions of a pest with its environment. The use of genetically characterized model plants can allow investigation of chosen aspects of these interactions by limiting the number of variables during experimentation. However, it is important to study the generic nature of these model systems if the data generated are to be assessed in a wider context, for instance, with those systems of commercial significance. This study assesses the suitability of Arabidopsis thaliana (L.) Heynh. (Brassicaceae) as a model host plant to investigate plant-herbivore-natural enemy interactions, with Plutella xylostella (L.) (Lepidoptera: Plutellidae), the diamondback moth, and Cotesia plutellae (Kurdjumov) (Hymenoptera: Braconidae), a parasitoid of P. xylostella. The growth and development of P. xylostella and C. plutellae on an A. thaliana host plant (Columbia type) were compared to that on Brassica rapa var. pekinensis (L.) (Brassicaceae), a host crop that is widely cultivated and also commonly used as a laboratory host for P. xylostella rearing. The second part of the study investigated the potential effect of the different A. thaliana background lines, Columbia and Landsberg (used in wider scientific studies), on growth and development of P. xylostella and C. plutellae. Plutella xylostella life history parameters were found generally to be similar between the host plants investigated. However, C. plutellae were more affected by the differences in host plant. Fewer adult parasitoids resulted from development on A. thaliana compared to B. rapa, and those that did emerge were significantly smaller. Adult male C. plutellae developing on Columbia were also significantly smaller than those on Landsberg A. thaliana.

Mortality of Plutella xylostella (Lepidoptera, Plutellidae) by parasitoids in the Province of Santa Fe, Argentina

Mortality of Plutella xylostella (Lepidoptera, Plutellidae) by parasitoids in the Province of Santa Fe, Argentina. Plutella xylostella (Linnaeus, 1758) (Lepidoptera, Plutellidae) larvae cause severe economic damage on cabbage, Brassica oleracea L. variety capitata (Brassicaceae), in the horticultural fields in the Province of Santa Fe, Argentina. Overuse of broad spectrum insecticides affects the action of natural enemies of this insect on cabbage. The objectives of this work were to identify the parasitoids of P. xylostella and to determine their influence on larva and pupa mortality. Weekly collections of larvae and pupae were randomly conducted in cabbage crops during spring 2006 and 2007. The immature forms collected were classified according to their developmental stage: L1 and L2 (Ls = small larvae), L3 (Lm = medium larvae), L4 (Ll = large larvae), pre-pupae and pupae (P). Each individual was observed daily in the laboratory until the adult pest or parasitoid emergence. We identified parasitoids, the number of instar and the percentage of mortality of P. xylostella for each species of parasitoid. Parasitoids recorded were: Diadegma insulare (Cresson, 1875) (Hymenoptera, Ichneumonidae), Oomyzus sokolowskii (Kurdjumov, 1912) (Hymenoptera, Eulophidae), Cotesia plutellae (Kurdjumov, 1912) (Hymenoptera, Braconidae) and an unidentified species of Chalcididae (Hymenoptera). Besides parasitoids, an unidentified entomopathogenic fungus was also recorded in 2006 and 2007. In 2006, the most successful parasitoids were D. insulare and O. sokolowskii, while in 2007 only D. insulare exerted a satisfactory control and it attacked the early instars of the pest.

Lack of intraspecific biological variation between two geographical populations of Oomyzus sokolowskii (Hymenoptera: Eulophidae), a gregarious larval–pupal parasitioid of Plutella xylostella (Lepidoptera: Plutellidae)

The chalcid, Oomyzus sokolowskii Kurdjumov has been recorded in many parts of the world as a major larval-pupal, gregarious endoparasitoid of the diamondback moth, Plutella xylostella (Linnaeus), a serious pest of brassica vegetable crops worldwide. This study investigated intraspecific variation between two populations of O. sokiolowskii, one from Cape Verde Islands, West Africa and the other from Hangzhou, China. In all crosses and backcrosses between the two geographical populations, the numbers of progeny and sex ratio of progeny were similar to those obtained within each of the populations, demonstrating complete reproductive compatibility between the two populations. The two populations showed similar responses to temperature with respect to development time and survival of immature stages. Observations on the interactions between the two O. sokolowskii populations and Cotesia plutellae (Kurdjumov), another major parasitoid of P. xylostella, showed that neither population could achieve successful parasitism of P. xylostella larvae already parasitized by C. plutellac. However, both O. sokolowskii populations could achieve hyperparasitism by ovipositing into a mid-late stage larva of C. plutellae developing inside the primary host. Contrary to earlier reports, no evidence of intraspecific variations in ability to hyperparasitize between these two populations of O. sokolowskii was found.

Inorganic elements in the fat bodies of Diatraea saccharalis (Lepidoptera: Crambidae) larvae parasitized by Cotesia flavipes (Hymenoptera: Braconidae)

Koinobiont parasitoids use several strategies to regulate the host`s physiological processes during parasitism. Although many aspects of host-parasitoid interactions have been explored, studies that attempted to assess the effects of parasitism on the availability of inorganic elements in the host are virtually nonexistent. Therefore, we aimed to evaluate the effects of parasitism on the concentrations of inorganic elements in the fat bodies of larvae of Diatraea saccharalis (Lepidoptera: Crambidae) during the development of the parasitoid Cotesia flavipes (Hymenoptera: Braconidae), by using total reflection X-ray fluorescence (TXRF). TXRF analysis allowed comparisons of the changes in the availability of the elements P. S. K, Ca, Cr, Fe, Ni, Cu, and Zn in the fat body tissues of D. saccharalis larvae parasitized by C. flavipes. Overall, the concentration of inorganic elements was higher early in parasitoid development (1 and 3 days after parasitism) compared to non-parasitized larvae, but much lower towards the end of parasitoid development (7 and 9 days after parasitism). Ca, K, and S were reduced after the fifth day of parasitism, which affected the total abundance of inorganic elements observed in the fat bodies of the parasitized hosts. The regulatory mechanisms or pathological effects related to the observed variation of the host inorganic elements induced by the parasitoid remain unknown, but there might be a strategy to make these elements available to the parasitoid larvae at the end of their development, when higher metabolic activity of the host fat body is required to sustain parasitoid growth. The observed variation of the host`s inorganic elements could also be related to the known effects of parasitism on the host`s immune response. (C) 2010 Elsevier Inc. All rights reserved.

Changes in the hemolymph and fat body metabolites of Diatraea saccharalis (Fabricius) (Lepidoptera : Crambidae) parasitized by Cotesia flavipes (Cameron) (Hymenoptera : Braconidae)

The koinobiont Cotesia flavipes responds to and is influenced by biochemical changes in the host hemolymph composition, Diatraea saccharalis. Changes in the composition of macronutrients may occur due to the hosts own development or by changes induced after parasitization. These changes occur to facilitate parasitoid invasion and to make the host internal environment suitable to parasitoid immature development. Therefore, changes in the availability of stored and circulating nutrients may correlate with the nutritional requirements of specific parasitoid immature stages. In here, we describe changes in the biochemical composition of parasitized and control larvae at different stages of parasitoid development to gain information on C flavipes host regulation and on its quantitative immature nutritional requirements. Total proteins, lipids and carbohydrates free in the hemolymph or stored in host fat bodies, and the SDS-PAGE protein profile of the hemolymph were evaluated in control and parasitized 6th instar during the whole parasitoid development. Changes in the total protein available in the host hemolymph were detected soon after parasitization, but carbohydrate and lipids were observed to differ only towards parasitoid larvae egression. Although C. flavipes affected the availability of all macronutrients observed in the host hemolymph, lipids and proteins stored in the host fat bodies were unaffected. However, carbohydrate concentration at the end of parasitoid larval development was much lower in parasitized than in control larvae at the same stage of development. SDS-PAGE analysis indicated C flavipes up-regulated two host proteins (125 and 48 kDa) and released two parasitism-specific proteins towards the end of parasitoid larval development. We provide a discussion on the role these changes may have on the process of host regulation and their possible requirement to sustain parasitoid development. (C) 2007 Elsevier Inc. All rights reserved.

Promoter studies of a polydnavirus gene from Cotesia rubecula (Hym : Braconidae)

The Cotesia rubecula polydnavirus gene, CrV1, is expressed in a highly transient fashion. Within four hours after egg deposition and virus infection, tissues of the host caterpillar, Pieris rapae, express high levels of the transcript. Twelve hours after infection no transcripts are visible. We have previously shown that the CrV1 secreted protein is mainly produced in host haemocytes. In haemocytes, immune functions such as phagocytosis and cell spreading are abolished by destabilization of the cell cytoskeleton. To test whether the observed down-regulation of CrV1 transcripts is mediated by transcriptional control or by other factors, such as the disruption of cytoskeleton in CrV1-inactivated cells, we cloned the promoter and the 3' untranslated region of the CrV1 gene to study CrV1 expression. The promoter region of the CrV1 gene was cloned into baculovirus expression systems along with the CAT reporter gene. Molecular analyses showed that the CAT gene under the control of CrV1 promoter is expressed as early as 2 h post infection and continues until late phase of infection suggesting that down-regulation of CrV1 expression in host haemocytes is perhaps mediated by post-transcriptional mechanisms.

Characterization of a novel protein with homology to C-type lectins expressed by the Cotesia rubecula bracovirus in larvae of the lepidopteran host, Pieris rapae

Polydnaviruses are essential for the survival of many Ichneumonoid endoparasitoids, providing active immune suppression of the host in which parasitoid larvae develop. The Cotesia rubecula bracovirus is unique among polydnaviruses in that only four major genes are detected in parasitized host ( Pieris rapae) tissues, and gene expression is transient. Here we describe a novel C. rubecula bracovirus gene (CrV3) encoding a lectin monomer composed of 159 amino acids, which has conserved residues consistent with invertebrate and mammalian C-type lectins. Bacterially expressed CrV3 agglutinated sheep red blood cells in a divalent ion-dependent but Ca2+-independent manner. Agglutination was inhibited by EDTA but not by biological concentrations of any saccharides tested. Two monomers of similar to14 and similar to17 kDa in size were identified on SDS-PAGE in parasitized P. rapae larvae. The 17-kDa monomer was found to be an N-glyscosylated form of the 14-kDa monomer. CrV3 is produced in infected hemocytes and fat body cells and subsequently secreted into hemolymph. We propose that CrV3 is a novel lectin, the first characterized from an invertebrate virus. CrV3 shows over 60% homology with hypothetical proteins isolated from polydnaviruses in two other Cotesia wasps, indicating that these proteins may also be C-type lectins and that a novel polydnavirus lectin family exists in Cotesia-associated bracoviruses. CrV3 is probably interacting with components in host hemolymph, resulting in suppression of the Pieris immune response. The high similarity of CrV3 with invertebrate lectins, as opposed to those from viruses, may indicate that some bracovirus functions were acquired from their hosts.

Isolation and characterization of a novel venom protein from an endoparasitoid, Cotesia rubecula (Hym : Braconidae)

Insects are important vectors of diseases with remarkable immune defense capabilities. Hymenopteran endoparasitoids are adapted to overcome the host defense system and, therefore, are useful sources of immune-suppressing proteins. Not much is known about venom proteins in endoparasitoids, especially those that have a functional relationship with polydnaviruses (PDVs). Here, we describe the isolation and characterization of a small venom protein (Vn4.6) from an endoparositoid, Cotesia rubecula, which interferes with the activation of the host hemolymph prophenoloxidose. The coding region for Vn4.6 is located upstream in the opposite direction of a gene coding for a C rubecula PDV-protein (Crp32). Arch. Insect Biochem. Physiol. 53:92-100, 2003. (C) 2003 Wiley-Liss, Inc.

Persistence and expression of Cotesia congregata polydnavirus in host larvae of the tobacco hornworm, Manduca sexta

The gregarious braconid wasp Cotesia congregata parasitizes host larvae of Manduca sexta, and several other sphingid species. Parasitism induces host immunosuppression due to the disruptive action of the wasp's polydnavirus (PDV) on host blood cells. During the initial stages of parasitism, these cells undergo apoptosis followed by cell clumping, which clears the hemolymph of a large number of cells. In this study, the persistence and expression of Cotesia congregata PDV (CcPDV) were examined using Southern and Nor-them blots, respectively. Digoxygenin-labelled total polydnaviral DNA was used to probe genomic DNA isolated from fat body and brains of hosts with emerged wasps taken 6 days following egress of the parasitoids, and significant cross-hybridization between the host fat body genomic DNA with viral DNA was seen. Thus, the virus persists in the host for the duration of parasitism. even during the post-emergence period, and may even be integrated in the host caterpillar DNA. Viral gene expression was examined using Northern blots and probes to the Cotesia rubecula CrV1 homolog, and the CrV1-like mRNAs were expressed as early as 4 h post-parasitization for at least 72 h and faint hybrization is even seen at the time the wasps eclose. In contrast, in Pieris rapae larvae the CrV1 transcript is expressed only for a brief time, during which time hemocyte function is disrupted. The effect is transitory, and hemocytes regain their normal functions after the parasites emerge as first instars. The genome of CcPDV contains one copy of the CrV1-like homolog as shown on Southern blots of viral genomic DNA. In conjunction with our earlier studies of the PDV-encoded early protein 1, the current work suggests multiple viral transcripts are produced following parasitization of the host. and likely target host hemocytes to induce their apoptosis, thereby preventing encapsulation of the parasitoid's eggs. Whether viral DNAs are integrated in the host's genomic DNA remains to be proven, but our results provide preliminary evidence that viral DNAs are detected in the host's fat body cells examined at the time of wasp ernergence and several days later. (C) 2003 Elsevier Science Ltd. All rights reserved.

The effect of resource aggregation at different scales: Optimal foraging behavior of Cotesia rubecula

Resources can be aggregated both within and between patches. In this article, we examine how aggregation at these different scales influences the behavior and performance of foragers. We developed an optimal foraging model of the foraging behavior of the parasitoid wasp Cotesia rubecula parasitizing the larvae of the cabbage butterfly Pieris rapae. The optimal behavior was found using stochastic dynamic programming. The most interesting and novel result is that the effect of resource aggregation within and between patches depends on the degree of aggregation both within and between patches as well as on the local host density in the occupied patch, but lifetime reproductive success depends only on aggregation within patches. Our findings have profound implications for the way in which we measure heterogeneity at different scales and model the response of organisms to spatial heterogeneity.

Optimal patch-leaving behaviour: a case study using the parasitoid Cotesia rubecular

1. Parasitoids are predicted to spend longer in patches with more hosts, but previous work on Cotesia rubecula (Marshall) has not upheld this prediction, Tests of theoretical predictions may be affected by the definition of patch leaving behaviour, which is often ambiguous. 2. In this study whole plants were considered as patches and assumed that wasps move within patches by means of walking or flying. Within-patch and between-patch flights were distinguished based on flight distance. The quality of this classification was tested statistically by examination of log-survivor curves of flight times. 3. Wasps remained longer in patches with higher host densities, which is consistent with predictions of the marginal value theorem (Charnov 1976). tinder the assumption that each flight indicates a patch departure, there is no relationship between host density and leaving tendency. 4. Oviposition influences the patch leaving behaviour of wasps in a count down fashion (Driessen et al. 1995), as predicted by an optimal foraging model (Tenhumberg, Keller & Possingham 2001). 5. Wasps spend significantly longer in the first patch encountered following release, resulting in an increased rate of superparasitism.

Preferência de Cotesia flavipes (Cameron, 1891) (Hymenoptera: Braconidae) por lagartas de Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) alimentadas com diferentes cultivares de cana-de-açúcar

O objetivo desta pesquisa foi avaliar a preferência do parasitoide Cotesia flavipes (Hymenoptera: Braconidae) por lagartas de Diatraea saccharalis (Lepidoptera: Crambidae), alimentadas com diferentes cultivares de cana-de-açúcar. O experimento foi conduzido em laboratório, em duas condições de alimentação, uma envolvendo lagartas de D. saccharalis, alimentadas em dieta artificial, e, a outra, com lagartas alimentadas em dieta artificial e mantidas temporariamente nos toletes dos cultivares de cana-de-açúcar. Os cultivares utilizados foram: SP80-1842 e SP81-3250, resistentes, e RB855536, suscetível à D. saccharalis. A preferência para oviposição das fêmeas de C. flavipes foi avaliada em testes com e sem chance de escolha, em delineamento inteiramente casualizado, com quatro tratamentos e 15 repetições. Lagartas de D. saccharalis com 19 dias de idade foram oferecidas para fêmeas de C. flavipes, acasaladas e com 24 horas de idade, em ambos os testes. No teste com chance de escolha, utilizou-se um olfatômetro, o qual constou de quatro compartimentos, em cujo centro liberaram-se quatro fêmeas de C. flavipes, enquanto, no teste sem chance de escolha, foram utilizadas placas de Petri, no interior das quais se colocou uma lagarta de D. saccharalis, oriunda das duas condições de alimentação. Nestes testes, foi observada a percentagem de parasitismo e, continua-mente, o comportamento da primeira escolha de fêmeas de C. flavipes, em intervalos de zero a um, um a três, três a cinco, cinco a oito e de oito a dez minutos após as liberações. Lagartas de D. saccharalis, oriundas da alimentação em dieta artificial com colmos triturados dos cultivares, foram igualmente preferidas para atratividade de fêmeas do parasitoide C. flavipes. A percentagem de parasitismo de lagartas de D. saccharalis, criadas com dietas artificiais com colmos dos cultivares SP80-1042 e RB855536, foi igualmente parasitadas por C. flavipes.

Organic and conventional fertilizer effects on a tritrophic interaction: parasitism, performance and preference of Cotesia vestalis

Interest in sustainable farming methods that rely on alternatives to conventional synthetic fertilizers and pesticides is increasing. Sustainable farming methods often utilize natural populations of predatory and parasitic species to control populations of herbivores, which may be potential pest species. We investigated the effects of several types of fertilizer, including those typical of sustainable and conventional farming systems, on the interaction between a herbivore and parasitoid. The effects of fertilizer type on percentage parasitism, parasitoid performance, parasitoid attack behaviour and responses to plant volatiles were examined using a model Brassica system, consisting of Brassica oleracea var capitata, Plutella xylostella (Lepidoptera) larvae and Cotesia vestalis (parasitoid). Percentage parasitism was greatest for P. xylostella larvae feeding on plants that had received either a synthetic ammonium nitrate fertilizer or were unfertilized, in comparison to those receiving a composite fertilizer containing hoof and horn. Parasitism was intermediate on plants fertilized with an organically produced animal manure. Male parasitoid tibia length showed the same pattern as percentage parasitism, an indication that offspring performance was maximized on the treatments preferred by female parasitoids for oviposition. Percentage parasitism and parasitoid size were not correlated with foliar nitrogen concentration. The parasitoids did not discriminate between hosts feeding on plants in the four fertilizer treatments in parasitoid behaviour assays, but showed a preference for unfertilized plants in olfactometer experiments. The percentage parasitism and tibia length results provide support for the preference–performance hypothesis

Quality of Cotesia flavipes (Hymenoptera: Braconidae) reared at different host densities and the estimated cost of its commercial production

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Fonte proteica na criação de Diatraea saccharalis e seu reflexo na produção e no controle de qualidade de Cotesia flavipes

O objetivo do trabalho foi avaliar diferentes dietas para lagartas de Diatraea saccharalis, com o intuito de obter informações sobre as características biológicas da praga e consequentemente avaliar a qualidade do parasitoide Cotesia flavipes, visando melhorar tecnicamente sua produção massal. Lagartas de 24 horas de idade foram transferidas para tubos (25 lagartas/tubo) contendo dieta artificial com levedura de cerveja e germe de trigo e outra somente com germe de trigo como fonte de proteína. Depois de aproximadamente 15 dias, as lagartas foram retiradas dos tubos e acondicionadas em placas com dieta, sendo observadas lagartas de D. saccharalis não parasitadas e parasitadas para avaliação das características biológicas de D. saccharalis e C. flavipes.nas diferentes dietas. Além da observação das características biológicas de D. saccharalis, foram realizadas medições nas lagartas de 15 dias de idade. Também, massas de pupas de C. flavipes.produzidas em biofábrica foram classificadas em três diferentes tamanhos, sendo pequenas (1,3 a 2,5 cm de comprimento), médias (2,5 a 3,5 cm de comprimento) e grandes (3,5 a 4,0 cm de comprimento). Após a emergência dos adultos foi realizado o parasitismo em lagartas de D. saccharalis, sendo observados o número de machos e fêmeas, tamanho da massa, razão sexual e número de pupas inviáveis. A dieta artificial que contém somente germe de trigo é a mais indicada para criação massal do hospedeiro D. saccharalis para produção em larga escala de C. flavipes. Massas de pupas classificadas como grandes possuem melhor qualidade em criações massais de C. flavipes.