Giovani. Variabilidade de parâmetros biológicos e genéticos de Cotesia flavipes (Cameron, 1891) (Hymenoptera: Braconidae) no Brasil.

2016

A protein with protective properties against the cellular defense reactions in insects

The molecular mechanism of how insects recognize intruding microorganisms and parasites and distinguish them from own body structures is not well known. We explored evolutionary adaptations in an insect parasitoid host interaction to identify components that interfere with the recognition of foreign objects and cellular encapsulation. Because some parasitoids provide protection for the developing wasp in the absence of an overt suppression of the insect host defense, we analyzed the surface of eggs and symbiotic viruses for protective properties. Here we report on the molecular cloning of a 32-kDa protein (Crp32) that is one of the major protective components. It is produced in the calyx cells of the female wasp ovaries and attached to the surface of the egg and other particles including polydnaviruses. The recombinant protein confers protection to coated objects in a cellular encapsulation assay suggesting that a layer of Crp32 may prevent cellular encapsulation reactions by a local inactivation of the host defense system.

A coiled-coil region of an insect immune suppressor protein is involved in binding and uptake by hemocytes

Polydnaviruses are associated with certain parasitoid wasps and are introduced into the body cavity of the host caterpillar during oviposition. Some of the viral genes are expressed in host tissues and corresponding proteins are secreted into the hemocoel causing suppression of the host immune system. The Cotesia rubecula polydnavirus gene product, CrV1, effectively inactivates hemocytes by mediating cytoskeleton break-down. A precondition for the CrV1 function is the incorporation of the extracellular protein by hemocytes. Here, we show that a coiled-coil domain containing a putative leucine zipper is required for CrV1 function, since removal of this domain abolishes binding and uptake of the CrV1 protein by hemocytes. (C) 2002 Elsevier Science Ltd. All rights reserved.

Polydnavirus particle proteins with similarities to molecular chaperones, heat-shock protein 70 and calreticulin

Multipartite nucleic acid-containing virus-like particles, known as polydnaviruses, are special structures produced by female parasitoid wasps to deliver wasp components into the body of their host at oviposition. The particles confer protection for the developing parasitoid by passive and active means. Although several genes expressed from the circular DNA of these particles have been identified from various host-parasitoid systems, there is not much known about the structural proteins of these particles. Here we report on two genes encoding Cotesia rubecula particle proteins with similarities to molecular chaperones, calreticulin and heat-shock protein 70.

Virus or not? Phylogenetics of polydnaviruses and their wasp carriers

Our current, still limited, understanding of the comparative biology and evolution of polydnaviruses (PDVs) is reviewed, especially in the context of the possible origins of these parasitoid viruses and of their coevolution with carrier wasps. A hypothetical scenario of evolution of PDVs from ascovirus (or ascovirus-like) ancestors is presented, with examples of apparent extant transitional forms. PDVs appear, in the case of bracoviruses, to show phylogenetic relationships that mirror those of their wasp carriers: with ichno-viruses, the picture is less clear. Ongoing sequencing studies of entire PDV genomes from diverse wasp species are likely to greatly contribute to our understanding of PDV evolution. (C) 2003 Elsevier Science Ltd. All rights reserved.

A serine proteinase homolog venom protein from an endoparasitoid wasp inhibits melanization of the host hemolymph

Activation of prophenoloxidase (proPO) in insects is a defense mechanism against intruding microorganisms and parasites. Pattern recognition molecules induce activation of an enzymatic cascade involving serine proteinases, which leads to the conversion of proPO to active phenoloxidase (PO). Phenolic compounds produced by pPO-activation are toxic to invaders. Here, we describe the isolation of a venom protein from the parasitoid, Cotesia rubecula, injected into the host, Pieris rapae, which is homologous to serine proteinase homologs (SPH). The data presented here indicate that the protein interferes with the proteolytic cascade, which under normal circumstances leads to the activation of proPO and melanin formation. (C) 2003 Elsevier Ltd. All rights reserved.

Differential effectiveness of microbially induced resistance against herbivorous insects in Arabidopsis.

Rhizobacteria-induced systemic resistance (ISR) and pathogen-induced systemic acquired resistance (SAR) have a broad, yet partly distinct, range of effectiveness against pathogenic microorganisms. Here, we investigated the effectiveness of ISR and SAR in Arabidopsis against the tissue-chewing insects Pieris rapae and Spodoptera exigua. Resistance against insects consists of direct defense, such as the production of toxins and feeding deterrents and indirect defense such as the production of plant volatiles that attract carnivorous enemies of the herbivores. Wind-tunnel experiments revealed that ISR and SAR did not affect herbivore-induced attraction of the parasitic wasp Cotesia rubecula (indirect defense). By contrast, ISR and SAR significantly reduced growth and development of the generalist herbivore S. exigua, although not that of the specialist P. rapae. This enhanced direct defense against S. exigua was associated with potentiated expression of the defense-related genes PDF1.2 and HEL. Expression profiling using a dedicated cDNA microarray revealed four additional, differentially primed genes in microbially induced S. exigua-challenged plants, three of which encode a lipid-transfer protein. Together, these results indicate that microbially induced plants are differentially primed for enhanced insect-responsive gene expression that is associated with increased direct defense against the generalist S. exigua but not against the specialist P. rapae.

Identification of parasitoid species using PCR amplification and restriction enzyme digestion

Spodoptera frugiperda is a pest of great economic importance in the Americas. It is attacked by several species of parasitoids, which act as biological control agents. Parasitoids are morphologically identifiable as adults, but not as larvae. Laboratory rearing conditions are not always optimal to rear out parasitic wasps from S. frugiperda larvae collected from wild populations, and it frequently happens that parasitoids do not complete their life cycle and stop developing at the larval stage. Therefore, we explored ways to identify parasitoid larvae using molecular techniques. Sequencing is one possible technique, yet it is expensive. Here we present an alternate, cheaper way of identifying seven species of parasitoids (Cotesia marginiventris, Campoletis sonorensis, Pristomerus spinator, Chelonus insularis, Chelonus cautus, Eiphosoma vitticolle and Meteorus laphygmae) using PCR amplification of COI gene followed by a digestion with a combination of four restriction endonucleases. Each species was found to exhibit a specific pattern when the amplification product was run on an agarose gel. Identifying larvae revealed that conclusions on species composition of a population of parasitic wasps can be biased if only the emerging adults are taken into account.

Parasitoids select plants more heavily infested with their caterpillar hosts: a new approach to aid interpretation of plant headspace volatiles

Plants produce volatile organic compounds (VOCs) in response to herbivore attack, and these VOCs can be used by parasitoids of the herbivore as host location cues. We investigated the behavioural responses of the parasitoid Cotesia vestalis to VOCs from a plant–herbivore complex consisting of cabbage plants (Brassica oleracea) and the parasitoids host caterpillar, Plutella xylostella. A Y-tube olfactometer was used to compare the parasitoids' responses to VOCs produced as a result of different levels of attack by the caterpillar and equivalent levels of mechanical damage. Headspace VOC production by these plant treatments was examined using gas chromatography–mass spectrometry. Cotesia vestalis were able to exploit quantitative and qualitative differences in volatile emissions, from the plant–herbivore complex, produced as a result of different numbers of herbivores feeding. Cotesia vestalis showed a preference for plants with more herbivores and herbivore damage, but did not distinguish between different levels of mechanical damage. Volatile profiles of plants with different levels of herbivores/herbivore damage could also be separated by canonical discriminant analyses. Analyses revealed a number of compounds whose emission increased significantly with herbivore load, and these VOCs may be particularly good indicators of herbivore number, as the parasitoid processes cues from its external environment

Campoletis flavicincta (Hym., Ichneumonidae) : ocorrência, criação e interação com Spodoptera frugiperda (Lep., Noctuidae) e Bacillus thuringiensis aizawai

Spodoptera frugiperda é um inseto-praga responsável por altos níveis de desfolhamento em gramíneas cultivadas, sendo que, dentre os métodos de controle, o biológico pode vir a tornar-se uma alternativa. Foi feita uma revisão de literatura sobre parasitóides de S. frugiperda. A ocorrência de parasitóides desse inseto, em áreas de cultivo de milho da EEA/IRGA, foi avaliada em Cachoeirinha, RS. Verificou-se a presença de Chelonus sp., Cotesia sp. e Exaticolus sp. (Hym., Braconidae), Campoletis flavicincta e Ophion sp. (Hym., Ichneumonidae) e de Archytas incertus e Lespesia archippivora (Dip., Tachinidae), com predomínio de C. flavicincta. Em função da dificuldade de obtenção de fêmeas deste inseto em laboratório, foram avaliadas diferentes condições de criação. Desta forma, registrou-se uma razão sexual de 0,41 quando foram expostas lagartas de segundo ínstar de S. frugiperda, as fêmeas do parasitóide apresentavam idade entre 3 e 6 dias e os casais foram formados no momento da exposição. Por fim, aspectos referentes à interação entre C. flavicincta/S. frugiperda/B. thuringiensis aizawai, em laboratório, foram avaliados A partir de análise do consumo alimentar de folhas de milho, observou-se que lagartas parasitadas e infectadas apresentaram um menor consumo, apesar do mesmo não ter diferido daquele de lagartas apenas parasitadas. A mortalidade das lagartas parasitadas e infectadas foi superior tanto das infectadas quanto das parasitadas. Lagartas infectadas mostraram um período de alimentação que não diferiu das sadias, apesar de terem apresentado maior duração da fase larval. Indivíduos descendentes de casais que emergiram de lagartas infectadas não tiveram alteradas suas características biológicas. A análise histológica de lagartas parasitadas e infectadas indicou não ter havido alteração no ovo e larva do parasitóide, resultante da ação do bacilo. Pode-se, portanto, inferir que o uso conjunto do parasitóide e da bactéria não resulta em prejuízo para o parasitóide.

Variabilidade espacial de populações de Diatraea saccharalis em canaviais e sugestão de método de amostragem

A broca Diatraea saccharalis é uma das mais importantes pragas da cana-de-açúcar no Brasil. O manejo de áreas infestadas está embasado no controle biológico com Cotesia flavipes, cujas liberações são feitas em função da população de lagartas da praga em campo. Por esta razão, a estimativa desta população é de fundamental importância para o sucesso do programa de manejo. Assim, o objetivo do presente trabalho foi estudar a variabilidade espacial de D. saccharalis em canaviais e sugerir um método de amostragem que permitisse uma estimativa adequada das populações em campo. Seis campos de aproximadamente 1 ha cada um foram amostrados, em grid retangular de aproximadamente 10 x 10 m, com ilhas. Cada ponto de amostragem foi representado pela coleta das formas biológicas da praga no interior dos colmos em 2 m de sulco. Na maioria das áreas, as populações de formas imaturas de D. saccharalis distribuíram-se de forma contagiosa, com alcance do semivariograma variando de 18 a 80 m. Com base no alcance médio, estimou-se em seis o número médio de pontos de amostragem por hectare que deveriam ser feitos para representar adequadamente a área. Este número foi menor do que o estimado utilizando parâmetros da estatística clássica. Sugere-se que, para estimativa das formas imaturas de D. saccharalis em canaviais, a amostragem seja feita em grid de 40 x 40 m, o que resulta em aproximadamente seis pontos de amostragem por hectare e permitirá, na maioria das áreas, efetuar levantamentos com independência e coeficientes de variação menores que 50%.

Resistência de genótipos de sorgo a Contarinia sorghicola (Coq.) (Diptera: Cecidomyiidae) e Diatraea saccharalis (Fabr.) (Lepidoptera: Pyralidae) e influência sobre parasitóides

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Possible causes of spatial variability in Diatraea saccharalis Fabr. (Lep., Crambidae) and its parasitoid populations in sugarcane fields in Brazil

Based on the large data set stored in some sugarcane mills regarding the biological control programme of Diatraea saccharalis using the parasitoids Cotesia flavipes and tachinid flies, the aim of the present study was to determine whether sugarcane varieties, number of cuts and plant ages are major determinants in promoting spatial variability in D. saccharalis, C. flavipes and tachinid fly populations in sugarcane fields in the state of São Paulo (Brazil). The data set used to support this study was provided by the Sao Joao and Barra sugarcane mills. Coefficient of variation was adopted as a measure of the spatial variability of population density. Spatial variability was estimated for the total density of D. saecharalis (parasitized + unparasitized larvae), and also for D. saccharalis larvae parasitized by tachinids and by C. flavipes. Statistical analysis revealed that the spatial variation in D. saccharalis populations was influenced by the number of cuts and by plant ages in the Barra and Sao Joao Mills, respectively. Similar results were obtained for D. saccharalis larvae parasitized by tachinids; however, in the Sao Joao Mill, the spatial variability of these populations was also influenced by the different number of varieties. Finally, considering D. saccharalis larvae parasitized by C. flavipes, no significant regressions with any of the three categories were observed for both sugarcane mills. The multitrophic implications for this agricultural system are also discussed.

Evidences of non-additive effects of multiple parasitoids on Diatraea saccharalis Fabr. (Lep., Crambidae) populations in sugarcane fields in Brazil

Biological control is a relatively benign method of pest control. However, considerable debate exists over whether multiple natural enemies often interact to produce additive or non-additive effects on their prey or host populations. Based on the large data set stored in the Sao Joao and Barra sugarcane mills (state of São Paulo, Brazil) regarding the programme of biological control of Diatraea saccharalis using the parasitoids Cotesia flavipes and tachinid flies, in the present study the author investigated whether the parasitoids released into sugarcane fields interfered significantly with the rate of parasitized D. saccharalis hosts. The author also observed whether there was an additive effect of releasing C. flavipes and tachinids on the rate of parasitized hosts, and looked for evidence of possible negative effects of the use of multiple parasitoid species in this biological control programme. Results showed that C. flavipes and the tachinids were concomitantly released in the Barra Mill, but not in the Sao Jao Mill. Furthermore, in the Barra Mill there was evidence that the parasitoids interacted because the percentage of parasitism did not increase after the release of either C. flavipes or tachinids. In the Sao Joao Mill, when both parasitoid species were released out of synchrony, both the percentage of parasitism by C. flavipes as well as that of the tachinids increased. When large numbers of tachinids were released in the Barra Mill, they caused a significant lower percentage of parasitism imposed by C. flavipes. The implications of the results as evidence of non-additive effects of C. flavipes plus tachinids on D. saccharalis populations are discussed.

The sugarcane borer Diatraea saccharalis (Fabr.) (Lep., Crambidae) and its parasitoids: a synchrony approach to spatial and temporal dynamics

A data set on Diatraea saccharalis and its parasitoids, Cotesia flavipes and tachinid flies, was analysed at five spatial scales-sugarcane mill, region, intermediary, farm and zone-to determine the role of spatial scale in synchrony patterns, and on temporal population variability. To analyse synchrony patterns, only the three highest spatial scales were considered, but for temporal population variability, all spatial scales were adopted. The synchrony-distance relationship revealed complex spatial structures depending on both species and spatial scale. Temporal population variability [SD log(x+1)] levels were highest at the smallest spatial scales although, in the majority of the cases, temporal variability was inversely dependent on sample size. All the species studied, with a few exceptions, presented spatial synchrony independent of spatial scale. The tachinid flies exhibited stronger synchrony dynamics than D. saccharalis and C. flavipes in all spatial scales with the latter displaying the weakest synchrony levels, except when mill spatial scales were compared. In some cases spatial synchrony may at first decay and then increase with distance, but the presence of such patterns can change depending on the spatial scale adopted.