Effects of female postmating odour on male sexual behaviour, in Heliconius butterflies /

The effects of the female postmating odour on male sexual behaviour were examined in Heliconius erato and H. charithonia (Lepidoptera: Nymphalidae). Predictions from the antiaphrodisiac hypothesis were tested using the two reproductive strategies of these species. Within the pupal mating strategy, results from behavioural experiments quantified and statistically tested dispersal rates of pupal-perched males to the presence of stimuli with and without the postmating odour. Results do not support an antiaphrodisiac function to the postmating odour. Similarly, within the adult courtship strategy, behavioural test results indicate that males do not alter their expenditure of energy in terms of either the duration or frequency of courtship behaviours elicited by females with and without the postmating odour. The data from both experiments did not support the antiaphrodisiac hypothesis for the function of the female postmating odour. A novel hypothesis predicting that the postmating female odour acts as an oviposition-deterring pheromone is presented.

Coevolution of Heliconius spp. and Passiflora spp. : a phylogenetic comparison

Although a substantial amount of research has been done on all aspects ofHeliconius biology and their ecological interactions with Passiflora, there has not hitherto been a phylogenetic examination of this association for coevolution. To test the HeliconiuslPassilfora association for coevolutionary congruence, phylogenies for each group were established and compared. The phylogeny for 14 species ofHeliconiinae from Costa Rica was based on combined sequence data from rRNA ITS 2 and partial EF-1a gene regions. For the Passifloraceae, 17 host plant species were utilized to establish a phylogeny based on tRNALeucine and ITS 1/5.8S1 ITS 2 sequence data. The phylogenies for both groups were largely in agreement with current classification (for Passifloraceae) and previously established phylogenies. Associations with the large subgenera Passiflora and Decaloba correspond with the two major Advanced Radiation groups in Heliconius. Although strict congruence above subgenus level was not observed, broad scale congruence was evident. One main host shift as well as other possible explanations for lack of strict congruence are suggested.

Genome-wide evidence for speciation with gene flow in Heliconius butterflies

Most speciation events probably occur gradually, without complete and immediate reproductive isolation, but the full extent of gene flow between diverging species has rarely been characterized on a genome-wide scale. Documenting the extent and timing of admixture between diverging species can clarify the role of geographic isolation in speciation. Here we use new methodology to quantify admixture at different stages of divergence in Heliconius butterflies, based on whole-genome sequences of 31 individuals. Comparisons between sympatric and allopatric populations of H. melpomene, H. cydno, and H. timareta revealed a genome-wide trend of increased shared variation in sympatry, indicative of pervasive interspecific gene flow. Up to 40% of 100-kb genomic windows clustered by geography rather than by species, demonstrating that a very substantial fraction of the genome has been shared between sympatric species. Analyses of genetic variation shared over different time intervals suggested that admixture between these species has continued since early in speciation. Alleles shared between species during recent time intervals displayed higher levels of linkage disequilibrium than those shared over longer time intervals, suggesting that this admixture took place at multiple points during divergence and is probably ongoing. The signal of admixture was significantly reduced around loci controlling divergent wing patterns, as well as throughout the Z chromosome, consistent with strong selection for Müllerian mimicry and with known Z-linked hybrid incompatibility. Overall these results show that species divergence can occur in the face of persistent and genome-wide admixture over long periods of time.

Genomic architecture of adaptive color pattern divergence and convergence in Heliconius butterflies

Identifying the genetic changes driving adaptive variation in natural populations is key to understanding the origins of biodiversity. The mosaic of mimetic wing patterns in Heliconius butterflies makes an excellent system for exploring adaptive variation using next-generation sequencing. In this study, we use a combination of techniques to annotate the genomic interval modulating red color pattern variation, identify a narrow region responsible for adaptive divergence and convergence in Heliconius wing color patterns, and explore the evolutionary history of these adaptive alleles. We use whole genome resequencing from four hybrid zones between divergent color pattern races of Heliconius erato and two hybrid zones of the co-mimic Heliconius melpomene to examine genetic variation across 2.2 Mb of a partial reference sequence. In the intergenic region near optix, the gene previously shown to be responsible for the complex red pattern variation in Heliconius, population genetic analyses identify a shared 65-kb region of divergence that includes several sites perfectly associated with phenotype within each species. This region likely contains multiple cis-regulatory elements that control discrete expression domains of optix. The parallel signatures of genetic differentiation in H. erato and H. melpomene support a shared genetic architecture between the two distantly related co-mimics; however, phylogenetic analysis suggests mimetic patterns in each species evolved independently. Using a combination of next-generation sequencing analyses, we have refined our understanding of the genetic architecture of wing pattern variation in Heliconius and gained important insights into the evolution of novel adaptive phenotypes in natural populations.

Influência do conteúdo nutricional de Passiflora suberosa e Passiflora misera na performance, nutrição, digestão e comportamento de escolha de oviposição de Heliconius erato phyllis (Lepidoptera : Nymphalidae)

Neste estudo, foram abordadas as implicações da condição nutricional e características físicas de Passiflora suberosa e Passiflora misera para a performance, comportamento de escolha pelo adulto, digestão e nutrição das larvas de H. erato phyllis. Inicialmente, avaliaram-se aspectos do desenvolvimento e foram realizadas medidas morfométricas de Passiflora suberosa e Passiflora misera cultivadas em três níveis de nitrogênio no solo. Avaliou-se o crescimento, a área foliar, o comprimento dos internódios, a dureza, a espessura, e a pilosidade, bem como analisou-se o conteúdo de água, de macro e micronutrientes presentes nos tecidos das respectivas plantas de ambas espécies. A seguir verificou-se a influência destas plantas na performance de Heliconius erato phyllis. Para isto, avaliou-se a sobrevivência, o tempo de desenvolvimento das larvas, o tamanho das pupas e dos adultos. Também, testou-se o comportamento de escolha dos adultos para oviposição, frente aos ramos de tais passifloráceas. Aspectos da ecologia nutricional das larvas foram avaliados, de forma comparativa, frente a P. suberosa e P. misera, através de índices nutricionais. Determinou-se: 1) o índice de consumo; 2) a taxa de crescimento relativo; 3) a digestibilidade aparente 4) a eficiência na conversão do alimento ingerido e 5) a eficiência na conversão do alimento digerido e convertido em biomassa. Ainda, 6) foi traçado um perfil de macro e micro nutrientes de P. suberosa e P. misera das plantas cultivadas e utilizadas para a criação das larvas, e de duas populações naturais. Em nível de digestão, determinou-se: 1) a quantidade de alimento ingerido, processado e retido 2) o tamanho dos fragmentos vegetais encontrados no intestino anterior e posterior; e ainda, 3) possíveis modificações morfológicas dos fragmentos, no decorrer da passagem destes através do tubo digestivo. O crescimento, a área foliar, e o comprimento dos internódios, de ambas passifloráceas, foram influenciados pelos níveis de nitrogênio. As plantas sem adição deste mineral apresentaram menores medidas em todos estes parâmetros. Foi detectada maior dureza e menor conteúdo de água nas primeiras folhas de P. suberosa cultivadas sem adição de nitrogênio. P. misera cultivada nesta condição também apresentou menor conteúdo de água, no entanto não houve maior dureza nas folhas. P.misera apresentou maior espessura nas primeiras e quintas folhas cultivadas com mais nitrogênio. A maioria dos macro e micronutrientes foram acumulados em maior concentração nas plantas com maior adição de nitrogênio no solo. O maior tempo no desenvolvimento e o menor tamanho alcançado pelos adultos foram causados pela menor porcentagem de nitrogênio e água contidos nos tecidos das plantas sem adição deste mineral. As larvas alimentadas com ramos de P. suberosa nesta condição, também enfrentaram a maior dureza da folha nos primeiros instares. As modificações morfológicas e nutricionais de ambas passifloráceas produziram efeitos na performance e na preferência de H. erato phyllis. A maior taxa de crescimento das larvas alimentadas com P. misera foi explicada pelo maior consumo e pela maior digestibilidade. Conseqüentemente, houve maior eficiência na conversão do alimento ingerido em substância corpórea. Não houve rompimentos das células da epiderme, nem dos fragmentos de P. misera nem dos de P. suberosa, retirados de ambas regiões intestinais. A maior permanência no intestino e o maior contato com o meio digestivo possibilitaram maior aproveitamento dos nutrientes de P. misera, em relação à P. suberosa. Isto ocorreu devido ao menor tamanho com que P. misera foi cortada pelas larvas, e pela menor espessura da folha. Quanto aos nutrientes, com exceção do fósforo e do manganês, os demais foram encontrados em maior concentração em P. suberosa. Dessa forma, as vantagens obtidas pelas larvas devem-se à maior eficiência destas no aproveitamento dos nutrientes contidos em P. misera, e não a um conteúdo nutricional superior nesta espécie.

Estretégias de acasalamento e qualidade do espermatóforo em heliconius (Lepidoptera, Nymphalidae)

Butterflies are insects known, in a variety of environments and for easy visual identification. The adult form may be frequently found in flowers looking for nectar. However, for many species of Heliconius (Lepidoptera, Nymphalidae) to visit the flower also represents the collecting of pollen, an important source of protein for adults. The protein obtained from the pollen allows the maintenance of physiological processes that increase the performance of the individual, promoting greater longevity and egg production. For males, proteins can also be part in your investment in reproductive success and fitness of offspring through a nutritional contribution that is transferred to the female in the act of mating as a nuptial present. It is known that this protein contains essential to the performance of the female oviposition, however the proportion of content and specific importance to the monogamous and polygamous species is not known yet. Whereas the species studied in this work have different patterns of mating in the strategy was to verify a significant difference in the quality of the spermatophore, and H. erato and H. melpomene, on the amount of protein present in this structure, indicating a difference in investment between the male reproductive strategies

A rede de marcação e recaptura: o caso de borboletas do gênero heliconius

(The Mark and Recapture Network: a Heliconius case study). The current pace of habitat destruction, especially in tropical landscapes, has increased the need for understanding minimum patch requirements and patch distance as tools for conserving species in forest remnants. Mark recapture and tagging studies have been instrumental in providing parameters for functional models. Because of their popularity, ease of manipulation and well known biology, butterflies have become model in studies of spatial structure. Yet, most studies on butterflies movement have focused on temperate species that live in open habitats, in which forest patches are barrier to movement. This study aimed to view and review data from mark-recapture as a network in two species of butterfly (Heliconius erato and Heliconius melpomene). A work of marking and recapture of the species was carried out in an Atlantic forest reserve located about 20km from the city of Natal (RN). Mark recapture studies were conducted in 3 weekly visits during January-February and July-August in 2007 and 2008. Captures were more common in two sections of the dirt road, with minimal collection in the forest trail. The spatial spread of captures was similar in the two species. Yet, distances between recaptures seem to be greater for Heliconius erato than for Heliconius melpomene. In addition, the erato network is more disconnected, suggesting that this specie has shorter traveling patches. Moving on to the network, both species have similar number of links (N) and unweighed vertices (L). However, melpomene has a weighed network 50% more connections than erato. These network metrics suggest that erato has more compartmentalized network and restricted movement than melpomene. Thus, erato has a larger number of disconnected components, nC, in the network, and a smaller network diameter. The frequency distribution of network connectivity for both species was better explained by a Power-law than by a random, Poissom distribution, showing that the Power-law provides a better fit than the Poisson for both species. Moreover, the Powerlaw erato is much better adjusted than in melpomene, which should be linked to the small movements that erato makes in the network

Estrutura genética populacional de Heliconius erato e Heliconius melpomene (Lepidoptera: Nymphalidae) em fragmentos de Mata Atlântica do Rio Grande do Norte

Extensive studies using molecular markers on butterflies have shown how a highly fragmented landscape may result in the reduction of gene flow among patches of habitat and, consequently, increase genetic differentiation among populations. However, little is known about Heliconius geographical structure and the effects of fragmentation on the connectivity of populations. Furthermore, findings on the effects of the population structure on the dynamics of mimicry evolution in Heliconius butterflies need to be tested in H. erato and H. melpomene specimens found in other locations other than Central and northern South Americas. For the present study, we had two motivations: (1) compare the population structure of H. erato and H. melpomene given the highly fragmented Brazil s Atlantic Forest habitat; and (2) studying population structure of co-mimics could give us insights into the dynamics of mimicry evolution. For this, we analysed the spatial structure and connectivity of eight populations of Heliconius butterflies, in a total of 137 H. erato specimens and 145 H. melpomene specimens, using nine microsatellites loci, 1144 AFLPs markers and 282 mitochondrial DNA sequences. In general, both species exhibited evidence of population subdivision but no isolation by distance indicating some extent of genetic differentiation among populations. Contrary to Kronforst & Gilbert s (2008) Costa Rican Heliconius, H. melpomene exhibited more genetic differentiation than H. erato based on nuclear markers. However, for mitochondrial DNA, H. erato populations showed more genetic differentiation than H. melpomene. Our results corroborate to other studies on Heliconius butterflies concerning the pronounced population subdivision and local genetic drift found in this genus. Nevertheless, the pattern of this differentiation varies significantly from the pattern found in studies conducted in Central America, where H. erato is generally more differentiated and structured than H. melpomene, based on nuclear markers. This different pattern may reflect different evolutionary histories of Heliconius species in Northeastern Brazil s Atlantic Forest

Biologia populacional comparada de duas espécies de heliconius (Lepidoptera: Nymphalidae) em um ambiente sazonal de floresta atlântica do nordeste brasileiro

Tropical environments often face strong seasonal variations in climate, such as alternate periods of dry and rain, that may often be important influence in the annual X the organisms lives. Here we assess how population dynamics of two butterfly species (Heliconius erato and Heliconius mepomene) respond to environmental and seasonal variations. A mark-release-recapture study carried out in an Atlantic forest reserve, 15 Km from Natal, Rio Grande do Norte, Brazil, for 3 years, during the dry and rainy season, with three visits weekly done. Information such as species, wing lenght, site of capture, pollen load and phenotype (number of spots) (in H. erato only) were noted for each capture. Seasonal variation exists in capture rates of the two species, with great capture rates during the rainy season. Despite finding differences in the mean density of individuals of the two species among the different collection areas, this difference was only significant between floodplain and central areas, and no influence of seasonality was observed in the mean density between the areas. Seasonality in wing size was only observed for H. erato, with larger wings during the rainy season. Females carried larger pollen loads than males both species, but species were similar. Only males differed seasonally, with larger pollen loads during the rainy season. The distribution of the number of wing spots did not vary between the dry and rainy seasons, and the number of spots in males and females was similar. Therefore, we conclude that there was a strong influence of seasonal variation in the population dynamic of the two Heliconius species, as well as in several aspects of their biology

Estratégias de acasalamento e qualidade do espermatóforo em heliconius (Lepidoptera, Nymphalidae)

Butterflies are insects known, in a variety of environments and for easy visual identification. The adult form may be frequently found in flowers looking for nectar. However, for many species of Heliconius (Lepidoptera, Nymphalidae) to visit the flower also represents the collecting of pollen, an important source of protein for adults. The protein obtained from the pollen allows the maintenance of physiological processes that increase the performance of the individual, promoting greater longevity and egg production. For males, proteins can also be part in your investment in reproductive success and fitness of offspring through a nutritional contribution that is transferred to the female in the act of mating as a nuptial present. It is known that this protein contains essential to the performance of the female oviposition, however the proportion of content and specific importance to the monogamous and polygamous species is not known yet. Whereas the species studied in this work have different patterns of mating in the strategy was to verify a significant difference in the quality of the spermatophore, and H. erato and H. melpomene, on the amount of protein present in this structure, indicating a difference in investment between the male reproductive strategies

A rede de marcação e recaptura: o caso de borboletas do gênero heliconius

(The Mark and Recapture Network: a Heliconius case study). The current pace of habitat destruction, especially in tropical landscapes, has increased the need for understanding minimum patch requirements and patch distance as tools for conserving species in forest remnants. Mark recapture and tagging studies have been instrumental in providing parameters for functional models. Because of their popularity, ease of manipulation and well known biology, butterflies have become model in studies of spatial structure. Yet, most studies on butterflies movement have focused on temperate species that live in open habitats, in which forest patches are barrier to movement. This study aimed to view and review data from mark-recapture as a network in two species of butterfly (Heliconius erato and Heliconius melpomene). A work of marking and recapture of the species was carried out in an Atlantic forest reserve located about 20km from the city of Natal (RN). Mark recapture studies were conducted in 3 weekly visits during January-February and July-August in 2007 and 2008. Captures were more common in two sections of the dirt road, with minimal collection in the forest trail. The spatial spread of captures was similar in the two species. Yet, distances between recaptures seem to be greater for Heliconius erato than for Heliconius melpomene. In addition, the erato network is more disconnected, suggesting that this specie has shorter traveling patches. Moving on to the network, both species have similar number of links (N) and unweighed vertices (L). However, melpomene has a weighed network 50% more connections than erato. These network metrics suggest that erato has more compartmentalized network and restricted movement than melpomene. Thus, erato has a larger number of disconnected components, nC, in the network, and a smaller network diameter. The frequency distribution of network connectivity for both species was better explained by a Power-law than by a random, Poissom distribution, showing that the Power-law provides a better fit than the Poisson for both species. Moreover, the Powerlaw erato is much better adjusted than in melpomene, which should be linked to the small movements that erato makes in the network

Diversidad de lepidópteros diurnos: Papilionidae, Pieridae y Nymphalidae en 11 hábitats de la zona núcleo de la reserva natural El Tisey, Estelí, Nicaragua, 2013

La Zona Núcleo de la Reserva el Tisey es atractiva para los turistas extranjeros y nacionales, debido a su diversidad bio-ecológica. Por ende se decidió caracterizar la diversidad de mariposas diurnas, ya que no existe información relacionadas de estas en la reserva para nuestro país. Se muestrearon once hábitats diferentes: Bosque de pino Santa Rosa, Bosque Mixto Napoleón, Bosque Ripario, Café Almaciguera, Café Jalacate, Camino, Potrero Zontul, Potrero Tisey Cerratos; con tres horas de captura y cuatro replicas por hábitats. Se utilizaron trampas Cebadas en hábitats Cerrados y red entomológica en hábitats abiertos, capturando un total de 2,516 especímenes distribuidas en 95 especies entre las familias Papilionidae, Pieridae y Nymphalidae. Las especies más abundantes fueron: Hermeuptychia hermes, Anartia fatima, Heliconius charithonius, Eurema salome jamapa y Eunica monima, Smyrna blomfildia y Eurema daira. La familia más predominante fue Nymphalidae (63% de las especies). La identificación se realizó con claves dicotómicas, pictográficas, taxonómicas y docentes del área de entomología. Los Gremios alimenticios fueron: Frugívoro, Nectarívoro, Frugívoro-Nectarívoro, Frugívoro-Coprófago y Nectarívoro-Coprófago, y hubo diferencia de especies por gremios, pero los gremios con mayor número de especies por hábitat son Nectarívoro (72%) y Frugívoro (13%). El método de colecta más eficiente fue la combinación de red entomológica y trampas para capturar especies de mariposas en la reserva El Tisey. Los hábitat más similares son Camino y Potrero Zontul compartiendo el 50% de las especies y los hábitat con menos similitud fueron Camino y Bosque Ripario, que resulto con el 44 %. Hubo diferencias en el número de especie y especímenes entre los hábitats en cuanto a la diversidad y abundancia, resultando una relación positiva entre la altura y la diversidad de mariposas.

Biotic versus abiotic pollination in Oeceoclades maculata (Lindl.) Lindl. (Orchidaceae)

Geographical variation in the reproductive biology of widespread species often occurs at their distributional boundaries. We sought to determine whether such variation has occurred in an invasive orchid, Oeceoclades maculata, across its naturalized range. We compared its reproductive biology in a Brazilian population with that published for a population on the Caribbean island of Puerto Rico. In the state of Sao Paulo, O. maculata flowers between December and February, at the height of the rainy season. Similar fruit sets were observed in manual self (76%) and cross (70.4%) pollination treatments. The fruit set of plants protected from both pollinators and rainfall was 6.1%, whereas plants exposed only to rainfall had a fruit set of 41.4%, slightly less than the controls (48.3%). Like the Puerto Rico population, reproduction is primarily through rain-assisted autogamy, but unlike observations made on the island, outcrossing can eventually occur. We observed two butterfly species (Heliconius ethilla narcaea and Heliconius erato phyllis) pollinating O. maculata. Secretory epidermal cells and trichomes of the spur lumen produced 0.7 mu L of 25% (sucrose equivalents) nectar per flower each morning, which was stored in a dilated basal portion of the spur and reabsorbed by the afternoon. Thus, geographical variation in reproductive biology exists across the broad invasive range of O. maculata.

What initiates speciation in passion-vine butterflies?

Studies of the continuum between geographic races and species provide the clearest insights into the causes of speciation. Here we report on mate choice and hybrid viability experiments in a pair of warningly colored butterflies, Heliconius erato and Heliconius himera, that maintain their genetic integrity in the face of hybridization. Hybrid sterility and inviability have been unimportant in the early stages of speciation of these two Heliconius. We find no evidence of reduced fecundity, egg hatch, or larval survival nor increases in developmental time in three generations of hybrid crosses. Instead, speciation in this pair appears to have been catalyzed by the association of strong mating preferences with divergence in warning coloration and ecology. In mate choice experiments, matings between the two species are a tenth as likely as matings within species. F1 hybrids of both sexes mate frequently with both pure forms. However, male F1 progeny from crosses between H. himera mothers and H. erato fathers have somewhat reduced mating success. The strong barrier to gene flow provided by divergence in mate preference is probably enhanced by frequency-dependent predation against hybrids similar to the type known to occur across interracial hybrid zones of H. erato. In addition, the transition between this pair falls at the boundary between wet and dry forest, and rare hybrids may also be selected against because they are poorly adapted to either biotope. These results add to a growing body of evidence that challenge the importance of genomic incompatibilities in the earliest stages of speciation.