Histological and electron microscopy observations on the testis and spermatogenesis of the butterfly Dione juno (Cramer, 1779) and Agraulis vanillae (Linnaeus, 1758) (Lepidoptera: Nymphalidae).

Lepidopteran species present an interesting case of sperm polymorphism and testicular fusion. The study of these features are of great importance in understanding the reproductive biology of these insects, especially in the case of those considered pests. Dione juno and Agraulis vanillae stand out as the most important pests of passion fruit (Passiflora sp.) crops in Brazil. Therefore, the objective of the present study was to characterize the testes and germ cells of Dione juno and Agraulis vanillae at different life stages, using light microscopy and scanning and transmission electron microscopy, to understand the maturation mechanisms of the male gametes in these species. The study showed that the larvae of both species have a pair of brown kidney-shaped testes, covered by epithelial cells which divide the organ into four follicles. The testes are full of spermatogonia which begin to differentiate in the third larval instar. In the fifth larval instar, spermatozoa can be observed. When they enter the prepupal stage the testes begin a fusion process that is completed in the adult insects, where they present as spherical organs divided into eight follicles, containing all the cells of the germ line. Spermatogenesis occurs centripetally, and in both species, sperm dimorphism is observed, where two different types of spermatozoa are formed, eupyrene (nucleated) and apyrene (anucleate), which differ in morphology and function. Apart from contributing to scientific basic research on the reproductive biology of these insects, the present study provides important data that can aid in research on the physiology, systematics, and control of these species.

Effect of Simulated Dasiops inedulis (Diptera: Lonchaeidae) Injury on Yield and Fruit Quality Parameters in Yellow Passionfruit.

Yellow passionfruit (Passiflora edulis f. flavicarpa O. Deg.) is a tropical fruit crop that is meeting increasing demand both in local and international markets in South America. The lance fly, Dasiops inedulis (Diptera: Lonchaeidae), affects P. edulis floral buds and flowers, and is thought to cause important yield losses in this crop. In Colombia, D. inedulis are commonly controlled through calendar-based applications of chemically synthesized insecticides, and no scientific criteria exist to guide pest management. In the present study, we simulated D. inedulis injury to passionfruit plants, over the course of three production cycles. We assessed the effect of seven different categories of flower bud removal (from 0% to 79.9%) on passionfruit yield and fruit quality parameters. Removal rates above 20% caused a significant reduction in the number of flowers, while yield levels were lowest at 50-79.9% bud removal. With increasing rates of flower bud removal, we observed higher initial production of buds and lower levels of natural abortion of floral and fruiting structures. For the three consecutive harvests, maximum yield levels were 7.57±5.51 kg (mean±SD; with 0-9.9% damage), and minimum yield was 2.37±2.15 kg (60-69.9% damage) per plant. For fruit quality parameters, D. inedulis injury did not affect fruit pulp weight or the content of soluble solids (Brix). Our work provides insights into the impact of D. inedulis on yellow passionfruit production, and constitutes a basis for future integrated pest management programs for this pest.

Ecological and genetic factors influencing the transition between host-use strategies in sympatric Heliconius butterflies.

Shifts in host-plant use by phytophagous insects have played a central role in their diversification. Evolving host-use strategies will reflect a trade-off between selection pressures. The ecological niche of herbivorous insects is partitioned along several dimensions, and if populations remain in contact, recombination will break down associations between relevant loci. As such, genetic architecture can profoundly affect the coordinated divergence of traits and subsequently the ability to exploit novel habitats. The closely related species Heliconius cydno and H. melpomene differ in mimetic colour pattern, habitat and host-plant use. We investigate the selection pressures and genetic basis underlying host-use differences in these two species. Host-plant surveys reveal that H. melpomene specializes on a single species of Passiflora. This is also true for the majority of other Heliconius species in secondary growth forest at our study site, as expected under a model of interspecific competition. In contrast, H. cydno, which uses closed-forest habitats where both Heliconius and Passiflora are less common, appears not to be restricted by competition and uses a broad selection of the available Passiflora. However, other selection pressures are likely involved, and field experiments reveal that early larval survival of both butterfly species is highest on Passiflora menispermifolia, but most markedly so for H. melpomene, the specialist on that host. Finally, we demonstrate an association between host-plant acceptance and colour pattern amongst interspecific hybrids, suggesting that major loci underlying these important ecological traits are physically linked in the genome. Together, our results reveal ecological and genetic associations between shifts in habitat, host use and mimetic colour pattern that have likely facilitated both speciation and coexistence.

Cytochrome P450-encoding genes from the Heliconius genome as candidates for cyanogenesis.

Cytochrome P450s are important both in the metabolism of xenobiotics and the production of compounds such as cyanogenic glucosides, which insects use in their defence. In the present study, we use transcriptomic and genomic information to isolate and name P450-encoding genes from the butterfly Heliconius melpomene. We classify each of the putative genes into its appropriate superfamily and compare the distribution of P450s across sequenced insects. We also identify homologues of two P450s known to be involved in cyanogenesis in the six-spot Burnet moth, Zygaena filipendulae. Classification of Heliconius P450s should be an important step in the dissection of their role in the exploitation of their host plant, the passion vine Passiflora.

Trail marking by caterpillars of the silverspot butterfly Dione juno huascuma.

A pheromone is implicated in the trail marking behavior of caterpillars of the nymphalid silverspot butterfly, Dione juno huascuma (Reakirt) (Lepidoptera: Heliconiinae) that feed gregariously on Passiflora (Malpighiales: Passifloraceae) vines in Mexico. Although they mark pathways leading from one feeding site to another with silk, this study shows that the silk was neither adequate nor necessary to elicit trail following behavior. Caterpillars marked trails with a long-lived pheromone that was deposited when they brushed the ventral surfaces of the tips of their abdomens along branch pathways. The caterpillars distinguished between pathways deposited by different numbers of siblings and between trails of different ages. Caterpillars also preferentially followed the trails of conspecifics over those of another nymphalid, Nymphalis antiopa L., the mourning cloak butterfly.

[Parasitism of passion fruit bugs in Brazil by Hexacladia smithii Ashmead (Hymenoptera: Encyrtidae)]. / Parasitismo de percevejos-praga do maracujazeiro no Brasil por Hexacladia smithii Ashmead (Hymenoptera: Encyrtidae).

We describe the new association of Hexacladia smithii (Ashmead) parasitizing two passion fruit bugs, Holhymenia histrio (Fabricius) and Anisoscelis foliacea marginella (Dallas) (Hemiptera: Coreidae), in Brazil.

Parasitismo de percevejos-praga do maracujazeiro no Brasil por Hexacladia smithii Ashmead (Hymenoptera: Encyrtidae) / Parasitism of passion fruit bugs in Brazil by Hexacladia smithii Ashmead (Hymenoptera: Encyrtidae)

We describe the new association of Hexacladia smithii (Ashmead) parasitizing two passion fruit bugs, Holhymenia histrio (Fabricius) and Anisoscelis foliacea marginella (Dallas) (Hemiptera: Coreidae), in Brazil.

First report and differential colonization of Passiflora Species by the B biotype of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) in Brazil / Primeiro relato e colonização diferencial de espécies de Passiflora pelo biótipo B de Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) no Brasil

This note is the first report of Bemisia tabaci (Gennadius) biotype B colonizing passionvine in Brazil. We examined the colonization of nine Passiflora species by a wild B type population under greenhouse conditions. P. amethystina Mikan was the most preferred species for oviposition and colonization, whereas P. suberosa L., P. coriacea Juss. and two commercially cultivated species, P. alata Curtis and P. edulis Sims f. flavicarpa Degener, were mostly uncolonised. P. morifolia Mast., P. cincinnata Mast., P. foetida L. and P. caerulea L. showed intermediate levels of colonization. Such differential colonization might suggest some degree of resistance by certain Passiflora species or oviposition preference by B. tabaci.

Esse trabalho descreve pela primeira vez a ocorrência do aleirodídeo Bemisia tabaci (Gennadius) biótipo B colonizando maracujazeiros no Brasil. Também foi examinada a colonização de nove espécies de Passiflora pelo inseto em condições de telado. P. amethystina Mikan foi a espécie de maior preferência para oviposição e colonização, enquanto P. suberosa L., P. coriacea Juss. e duas espécies cultivadas comercialmente, P. alata Curtis e P. edulis Sims f. flavicarpa Degener, foram pouco colonizadas pelo aleirodídeo. P. morifolia Mast., P. cincinnata Mast., P. foetida L. e P. caerulea L. exibiram níveis intermediários de colonização. Esses resultados sugerem que certas espécies de Passiflora exibem diferentes graus de resistência à colonização ou preferência para oviposição de B. tabaci biótipo B.

Feeding preference of Holymenia clavigera and Anisoscelis foliacea marginella (Hemiptera: Coreidae: Anisoscelini) in relation to intra- and interspecific features of host plants (Passifloraceae).

The interactions of Holymenia clavigera (Herbst) and Anisoscelis foliacea marginella (Dallas) with their hosts (Passifloraceae) in southern Brazil are poorly understood. Previous studies have indicated that both species use several host species and plant parts and have superior performance on wild hosts. This study evaluated feeding preferences and host plant use under both laboratory and field conditions. Choice tests were performed to determine preference for Passiflora suberosa parts, preference for P. suberosa and Passiflora misera immature and ripe fruits, and preference for P. suberosa, P. misera, and Passiflora edulis shoots. A field survey was carried out to assess H. clavigera and A. foliacea marginella use of P. suberosa parts. In addition, immature and ripe fruits of P. suberosa and P. misera were characterized in relation to pH, phenols, and anthocyanins. First-instar nymphs preferred the terminal buds, shifting to immature fruits in the second instar. Both coreids more frequently fed on immature fruits than on ripe ones. Neither species showed a significant preference among host species. The immature fruits presented higher phenol content as compared with the ripe fruits; the opposite was observed for anthocyanins. In the field, immature fruits and mature leaves were selected for feeding and resting. Thus, this study confirms the food mixing condition of these coreids, as well as the importance of fruits for their nutrition. Phenols, and presumably water, in the fruits may play a role in their feeding choices.

Herbivore handling of a Plants trichome: the case of Heliconius charithonia (L.) (Lepidoptera: Nymphalidae) and Passiflora lobata (Killip) Hutch. (Passifloraceae).

Trichomes reduce herbivore attack on plants by physically and/or chemically inhibiting movement or other activities. Despite evidence that herbivores are negatively affected by trichomes there also reports of insect counter-adaptations that circumvent the plants defense. This paper reports on a study that investigated the likely mechanisms employed by larvae of the nymphalid butterfly, Heliconius charithonia (L.), that allow it to feed on a host that is presumably protected by hooked trichomes (Passiflora lobata (Killip) Hutch). Evidence were gathered using data from direct observations of larval movement and behavior, faeces analysis, scanning electron microscopy of plant surface and experimental analysis of larval movement on plants with and without trichomes (manually removed). The latter involved a comparison with a non specialist congener, Heliconius pachinus Salvin. Observations showed that H. charithonia larvae are capable of freeing themselves from entrapment on trichome tips by physical force. Moreover, wandering larvae lay silk mats on the trichomes and remove their tips by biting. In fact, trichome tips were found in the faeces. Experimental removal of trichomes aided in the movement of the non specialist but had no noticeable effect on the specialist larvae. These results support the suggestion that trichomes are capable of deterring a non specialist herbivore (H. pachinus). The precise mechanisms that allow the success of H. charithonia are not known, but I suggest that a blend of behavioral as well as physical resistance mechanisms is involved. Future studies should ascertain whether larval integument provides physical resistance to trichomes.

First report and differential colonization of Passiflora species by the B biotype of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) in Brazil.

This note is the first report of Bemisia tabaci (Gennadius) biotype B colonizing passionvine in Brazil. We examined the colonization of nine Passiflora species by a wild B type population under greenhouse conditions. P. amethystina Mikan was the most preferred species for oviposition and colonization, whereas P. suberosa L., P. coriacea Juss. and two commercially cultivated species, P. alata Curtis and P. edulis Sims f. flavicarpa Degener, were mostly uncolonised. P. morifolia Mast., P. cincinnata Mast., P. foetida L. and P. caerulea L. showed intermediate levels of colonization. Such differential colonization might suggest some degree of resistance by certain Passiflora species or oviposition preference by B. tabaci.

Performance consequences of food mixing in two passion vine leaf-footed bugs, Holymenia clavigera (Herbst, 1784) and Anisoscelis foliacea marginella (Dallas, 1852) (Hemiptera; Coreidae).

Holymenia clavigera (Herbst) and Anisoscelis foliacea marginella (Dallas) (Hemiptera: Coreidae: Anisoscelini) are distributed in southern Brazil and use various passion vine species (Passifloraceae) as host-plants. Preliminary observations indicate a high coexistence of these species in terms of host-plant use; in addition, there is a strong similarity regarding egg and nymph morphology. In this study, the most suitable feeding sites for nymph performance on wild (Passiflora suberosa Linnaeus and Passiflora misera Humbold, Bonpland et Kunth) and cultivated (Passiflora edulis Sims) hosts were determined by rearing them on each host and on the combination of hosts. Performance was determined by evaluating nymph development and survivorship, and adult size at emergence. Plant parts used were also recorded. For both species, P. suberosa was the most suitable host plant. First instar nymphs of both species fed on terminal buds more frequently when compared to other plant parts. Second instar nymphs switched to green fruits, whose behavior was more pronounced for H. clavigera. Thus, H. clavigera and A. foliacea marginella immatures are extremely similar in terms of host-plant use and consequences for performance, in addition to their morphological similarity. We suggest that these coreids may have evolved through several processes, including parsimony between the immature stages after speciation, evolutionary convergence, mimicry or genetic drift.

Performance consequences of food mixing in two passion vine leaf-footed bugs, Holymenia clavigera (Herbst, 1784) and Anisoscelis foliacea marginella (Dallas, 1852) (Hemiptera; Coreidae)

Holymenia clavigera (Herbst) and Anisoscelis foliacea marginella (Dallas) (Hemiptera: Coreidae: Anisoscelini) are distributed in southern Brazil and use various passion vine species (Passifloraceae) as host-plants. Preliminary observations indicate a high coexistence of these species in terms of host-plant use; in addition, there is a strong similarity regarding egg and nymph morphology. In this study, the most suitable feeding sites for nymph performance on wild (Passiflora suberosa Linnaeus and Passiflora misera Humbold, Bonpland et Kunth) and cultivated (Passiflora edulis Sims) hosts were determined by rearing them on each host and on the combination of hosts. Performance was determined by evaluating nymph development and survivorship, and adult size at emergence. Plant parts used were also recorded. For both species, P. suberosa was the most suitable host plant. First instar nymphs of both species fed on terminal buds more frequently when compared to other plant parts. Second instar nymphs switched to green fruits, whose behavior was more pronounced for H. clavigera. Thus, H. clavigera and A. foliacea marginella immatures are extremely similar in terms of host-plant use and consequences for performance, in addition to their morphological similarity. We suggest that these coreids may have evolved through several processes, including parsimony between the immature stages after speciation, evolutionary convergence, mimicry or genetic drift.

Holymenia clavigera (Herbst) e Anisoscelis foliacea marginella (Dallas) (Hemiptera: Coreidae: Anisoscelini) ocorrem no sul do Brasil e são hóspedes de diversas espécies de Passifloraceae. Observações preliminares indicam uma alta coexistência destas espécies em termos de uso da planta hospedeira; em adição, ovos e ninfas são extremamente semelhantes. Neste estudo comparou-se a performance das ninfas das duas espécies em passifloráceas nativas silvestres (Passiflora suberosa Linnaeus e Passiflora misera Humbold, Bonpland et Kunth) e cultivadas (Passiflora edulis Sims), oferecidas em separado e conjuntamente. Foram determinadas as taxas de desenvolvimento e mortalidade ninfal, bem como o tamanho do adulto à emergência. As partes das hospedeiras mais utilizadas para alimentação foram registradas. Para ambos os coreídeos, P. suberosa conferiu melhor performance. Ninfas de primeiro instar de ambas as espécies alimentaram-se com maior intensidade da região apical dos ramos quando comparado com as demais partes das plantas. Ambas as espécies mudaram para os frutos verdes no segundo instar, sendo este comportamento mais pronunciado em H. clavigera. Deste modo, além da semelhança morfológica, os imaturos de H. clavigera and A. foliacea marginella são extremamente similares no tangente à interação com suas plantas hospedeiras. Sugere-se que estes coreídeos possam ter evoluído através de vários processos, entre os quais parcimônia entre as fases imaturas após especiação, convergência evolutiva, mimetismo ou deriva genética.

Effect of nitrogen on Passiflora suberosa L. (Passifloraceae) and consequences for larval performance and oviposition in Heliconius erato phyllis (Fabricius) (Lepidoptera: Nymphalidae)

Avaliou-se a influência do nitrogênio no crescimento e nas características morfológicas e nutricionais de Passiflora suberosa L. e as conseqüências no desempenho larval e oviposição de Heliconius erato phyllis (Fabricius). Foram utilizados três níveis de nitrogênio no solo (tratamentos) para o cultivo de P. suberosa: 0, 150 e 300 mg L-1. Larvas recém-eclodidas foram criadas em laboratório (25 ± 1ºC), individualmente, em ramos das plantas cultivadas em cada tratamento e fêmeas capturadas em campo foram submetidas a testes de escolha para oviposição. A taxa de crescimento, a área foliar e o comprimento dos internódios de P. suberosa aumentaram significativamente com a adição de nitrogênio. A dureza das folhas jovens nas plantas cultivadas sem adição de nitrogênio foi maior, as quais também apresentaram menor conteúdo de água. Houve maior concentração de nitrogênio e potássio nas plantas cultivadas com a adição de nitrogênio no solo. A qualidade nutricional do alimento expressa pela maior concentração de nitrogênio e água, diminuiu o tempo de desenvolvimento larval e aumentou o tamanho dos adultos, embora não tenha afetado a sobrevivência de forma significativa. As fêmeas escolheram os ramos das plantas fertilizadas para oviposição, o que foi positivamente relacionado com desempenho das larvas. Isto poderia explicar a manutenção desse comportamento, bem como a escolha por ramos maiores, registrado em estudos anteriores.

Transmissão do Passion fruit woodiness Virus por Aphis gossypii ( Glover ) ( Hemiptera: Aphididae ) e colonização de maracujazeiro pelo vetor / Transmission of Passion fruit woodiness virus by Aphis gossypii ( Glover ) ( Hemiptera: Aphididae ) and colonization of Passion flower by the vector

Avaliou-se a transmissão de Passion fruit woodiness virus (PWV) por Aphis gossypii (Glover). Em dois experimentos independentes, o afídeo transmitiu o PWV para maracujazeiros com taxas de 75 por cento e 100 por cento, ao se depositarem oito e doze afídeos virulíferos por planta, respectivamente. No final dos testes, observaram-se, em algumas plantas de maracujá, formas ápteras e ninfas de A. gossypii, sugerindo a colonização dessas plantas pelo afídeo. Esse parece ser o primeiro relato da colonização de Passiflora edulis f. flavicarpa (Deneger) por uma espécie de afídeo.

Effect of nitrogen on Passiflora suberosa L. (Passifloraceae) and consequences for larval performance and oviposition in Heliconius erato phyllis (Fabricius) (Lepidoptera: Nymphalidae).

The present study examines the influence of nitrogen on growth rates, morphological and nutritional characteristics of Passiflora suberosa L., and the corresponding consequences for Heliconius erato phyllis (Fabricius) growth and oviposition. The treatments consisted of different nitrogen doses (0, 150 and 300 mg L(-1)) applied to the soil. Newly emerged larvae were individually reared in the laboratory (25+/-1 degree C) on branches of P. suberosa obtained from plants grown under the three nitrogen levels, and field collected females were submitted to oviposition choice tests. P. suberosa growth rates, leaf area and internode length increased significantly when grown with nitrogen enriched soil. P. suberosa presented significantly greater leaf toughness when grown without nitrogen addition. Leaves of this species also presented lower water content and toughness. Nitrogen addition increased the concentration of nitrogen and potassium on plant leaf tissues. The greater nitrogen concentration on plants did not affect H. erato phyllis larval survival, but led to a reduction in larval development time and increased adult size. Females significantly preferred to lay eggs in plants cultivated in nitrogen enriched soil. This choice was positively correlated with larval performance, which could be a possible explanation for the maintenance of that behaviour, as well as the choice of larger branches as reported in previous studies.

[Transmission of Passion fruit woodiness virus by Aphis gossypii (Glover) (Hemiptera: Aphididae) and colonization of passion flower by the vector]. / Transmissão do Passion fruit woodiness virus por Aphis gossypii (Glover) (Hemiptera: Aphididae) e colonização de maracujazeiro pelo vetor.

The transmission of Passion fruit woodiness virus (PWV) by Aphis gossypii (Glover) was evaluated. In two independent experiments, A. gossypii transmitted PWV to passion fruit plants at the rates of 75% and 100%, when eight and twelve viruliferous aphids were deposited by plant, respectively. At the end of the tests, nymphs of A. gossypii were observed in some of the passion fruit plants, suggesting that the aphid species was colonizing the plants. This seems to be the first report of Passiflora edulis f. flavicarpa (Deneger) colonization by a species of aphid.

Geographical variation in larval host-plant use by Heliconius erato (Lepidoptera: Nymphalidae) and consequences for adult life history.

Adult body size, one of the most important life-history components, varies strongly within and between Heliconius erato phyllis (Lepidoptera: Nymphalidae) populations. This study determines if this variation is caused by geographical changes in host-plant used by the larval stage, whose reproductive parameters are influenced by female body size, with estimates of the corresponding heritability. The variation in adult body size was determined together with a survey of passion vine species (Passifloraceae) used by the larvae in seven localities in Rio Grande do Sul State: three located in the urban area of Porto Alegre and Triunfo Counties, two within Eucalyptus plantations (Barba Negra Forest, Barra do Ribeiro County, and Aguas Belas Experimental Station-Viamão County), one in a Myrtaceae Forest (Itapuã State Park-Itapuã County) and one in the Atlantic Rain Forest (Maquiné Experimental Station-Maquiné County). Effects of female body size on fecundity, egg size and egg viability were determined in an outdoor insectary. Size heritability was estimated by rearing in the laboratory offspring of individuals maintained in an insectary. The data showed that adults from populations where larvae feed only upon Passiflora suberosa are smaller than those that feed on Passiflora misera. The larvae prefer P. misera even when the dominant passion vine in a given place is P. suberosa. Fecundity increases linearly with the increase in size of females, but there is no size effect on egg size or viability. Size heritability is null for the adult size range occurring in the field. Thus, the geographical variation of H. erato phyllis adult size is primarily determined by the type, corresponding availability and quality of host-plants used by the larval stage. Within the natural size range of H. erato phyllis, the variation related to this character is not genetically based, thus being part of H. erato phyllis phenotypic plasticity.