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Background 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA), the main benzoxazinoid found in corn, elicits variable larval responses from different pest moths. For the widespread and highly polyphagous Spodoptera frugiperda (Lepidoptera: Noctuidae), the fall-armyworm (FAW), DIMBOA acts as a feeding stimulant and improves larval growth at low concentrations. The FAW present two host plant-related strains, corn and rice strains, related to host preference on corn and other Graminae or rice. Based on both host preference and strain divergence of the FAW on corn, a cereal containing DIMBOA, and rice, lacking this compound, we question if corn and rice strains larvae respond equally toward DIMBOA. We evaluated differential expression in the transcriptome of both midgut and fat body larval tissues of the two strains reared on either DIMBOA-enriched artificial diet or control diet and inferred Bayesian networks. Results We found differences in performance between corn and rice strain larvae reared on DIMBOA, as well as several differentially regulated contigs annotated as esterases, peptidases, transferases and reductases, all of them known for being related to responses of lepidopterans and other insects to DIMBOA. We also found a UDP-glucuronosyltransferase very similar to others found in many lepidopterans occupying a central hub within a transferase Bayesian network, suggesting that it is essential to an effective response to DIMBOA in FAW. Conclusion Our results suggest that there is an intrinsic cost for FAW rice strain larvae to metabolize corn-originated hydroxamic acids, which could have resulted in the partial host-associated genetic isolation found at FAW field populations.

Spodoptera frugiperda (J. E. Smith) is a serious and widespread agricultural pest with several records of resistance to different insecticides and Bt proteins, including the neurotoxic insecticides chlorpyrifos (organophosphate) and lambda-cyhalothrin (pyrethroid). In this study, we (i) characterized and monitored the susceptibility of field populations of S. frugiperda to chlorpyrifos (194 populations) and lambda-cyhalothrin (197 populations) collected from major maize-growing regions of Brazil from 2003 to 2016, and (ii) compared gene expression levels of chlorpyrifos- and lambda-cyhalothrin-resistant strains to a susceptible reference strain (Sf-ss) of S. frugiperda. Laboratory-guided assays to monitor larval susceptibility detected average survival ranging from 29.3% to 36.0% to chlorpyrifos, and 23.1% to 68.0% to lambda-cyhalothrin at diagnostic concentration, based on LC99 of the susceptible reference strain of each insecticide. The resistance ratio of the chlorpyrifos-resistant strain (Clo-rr) was 25.4-fold and of the lambda-cyhalothrin-resistant strain (Lam-rr) was 217-fold. Differential gene expression analyses between resistant vs susceptible strains identified 1,098 differentially expressed genes (DEGs) between Clo-rr and Sf-ss, and 303 DEGs between Lam-rr and Sf-ss. Functional analyses of the DEGs revealed the up-regulation of several detoxification enzymes, mainly cytochrome P450 belonging to the CYP3 and CYP6 clans. Genes associated with regulatory processes, such as the forkhead box O (FoxO) were also up-regulated. Our data points that the resistance mechanisms of Clo-rr and Lam-rr strains of S. frugiperda to chlorpyrifos and lambda-cyhalothrin are mainly mediated by enzyme detoxification.

The interdisciplinary field of Chemical Ecology in Brazil is currently composed of groups that emerged through the pioneering studies of Keith Spalding Brown Jr. and José Tércio Barbosa Ferreira. Following Keith Brown 's steps, José Roberto Trigo continued investigating the role of plant natural products in mediating the association among insects and their host plants, mainly in the Order Lepidoptera. The role of pyrrolizidine alkaloids in those associations was investigated extensively by Brown and Trigo, and most of what is currently known on this subject is based on their studies. The present work acknowledges their contribution to the Brazilian chemical ecology field and on insect-plant communication studies mediated by different chemical compounds.

Selection shapes genetic diversity around target mutations, yet little is known about how selection on specific loci affects the genetic trajectories of populations, including their genome-wide patterns of diversity and demographic responses. Here we study the patterns of genetic variation and geographic structure in a neotropical butterfly,Heliconius numata, and its closely related allies in the so-called melpomene-silvaniform clade.H. numatais known to have evolved an inversion supergene which controls variation in wing patterns involved in mimicry associations with distinct groups of co-mimics whereas it is associated to disassortative mate preferences and heterozygote advantage at this locus. We contrasted patterns of genetic diversity and structure 1) among extant polymorphic and monomorphic populations ofH. numata, 2) betweenH. numataand its close relatives, and 3) between ancestral lineages. We show thatH. numatapopulations which carry the inversions as a balanced polymorphism show markedly distinct patterns of diversity compared to all other taxa. They show the highest genetic diversity and effective population size estimates in the entire clade, as well as a low level of geographic structure and isolation by distance across the entire Amazon basin. By contrast, monomorphic populations ofH. numataas well as its sister species and their ancestral lineages all show lower effective population sizes and genetic diversity, and higher levels of geographical structure across the continent. One hypothesis is that the large effective population size of polymorphic populations could be caused by the shift to a regime of balancing selection due to the genetic load and disassortative preferences associated with inversions. Testing this hypothesis with forward simulations supported the observation of increased diversity in populations with the supergene. Our results are consistent with the hypothesis that the formation of the supergene triggered a change in gene flow, causing a general increase in genetic diversity and the homogenisation of genomes at the continental scale.

The global increase in species richness toward the tropics across continents and taxonomic groups, referred to as the latitudinal diversity gradient, stimulated the formulation of many hypotheses to explain the underlying mechanisms of this pattern. We evaluate several of these hypotheses to explain spatial diversity patterns in a butterfly family, the Nymphalidae, by assessing the contributions of speciation, extinction, and dispersal, and also the extent to which these processes differ among regions at the same latitude. We generate a time-calibrated phylogeny containing 2,866 nymphalid species (~45% of extant diversity). Neither speciation nor extinction rate variations consistently explain the latitudinal diversity gradient among regions because temporal diversification dynamics differ greatly across longitude. The Neotropical diversity results from low extinction rates, not high speciation rates, and biotic interchanges with other regions are rare. Southeast Asia is also characterized by a low speciation rate but, unlike the Neotropics, is the main source of dispersal events through time. Our results suggest that global climate change throughout the Cenozoic, combined with tropical niche conservatism, played a major role in generating the modern latitudinal diversity gradient of nymphalid butterflies., F.L.C is supported by an “Investissements d’Avenir” grant managed by the Agence Nationale de la Recherche (CEBA, ref. ANR-10-LABX-25-01). D.J.L. was supported by grants DEB-1541557 from NSF and WW-227R-17 from the National Geographic Society. R.I.V.W. was supported by Leverhulme Trust emeritus programme. M.E. was supported by an ATIP grant, a grant from the Human Frontier Science Program (RGP0014/2016) and a grant from the French National Research Agency (ANR CLEARWING ANR-16-CE02-0012). E.O.A. was supported by Sigma-Xi (G20100315153261), Center for Systematic Entomology and the Council of the Linnean Society and the Systematics Association for the Systematics Research Fund. S.N. was supported by the Swedish Research Council (2015-04218 and 2019-03441). P.M.M. was supported by a Czech Science Foundation grant (Junior GAČR, 20-18566Y). R.V. was supported by projects PID2019-107078GB-I00 / AEI / 10.13039/501100011033 (Agencia Estatal de Investigación) and 2017-SGR-991 (Generalitat de Catalunya). N.W. acknowledges support by the Swedish Research Council (2015-04441) and a start-up grant from the Department of Biology, Lund University.

Made available in DSpace on 2021-06-25T10:55:53Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Aim: There is little consensus on which environmental variables are best at predicting multiple dimensions of diversity. We ask whether there are common environmental correlates of diversity, despite ecological differences, across nine clades of plants and animals distributed along a single rainforest domain. For that, we compare the environmental correlates of species richness, phylogenetic diversity, and phylogenetic endemism. Location: Brazilian Atlantic Forest. Taxon:Five clades of plants (Bromelioideae, Miconieae, Bertolonia, Cambessedesieae, and the Fridericia and allies) and four clades of animals (butterlies in the tribe Ithomiini, frogs in the genera Boana and Proceratophrys, and birds in the subfamily Thraupinae). Methods: Using curated occurrence localities and phylogenetic data, we generated maps of (a) species richness, (b) phylogenetic diversity, (c) residuals of phylogenetic diversity regressed on species richness, and (d) phylogenetic endemism for all groups. We also compiled a set of 30 environmental descriptors, including records of current temperature and precipitation, climatic stability over time, and topography. Through a machine learning framework, we explored the environmental correlates of each of these diversity measures for each group. Results: The environmental variables used here were strong predictors of diversity for all studied groups. However, models for phylogenetic endemism had lower predictive power. Although patterns of diversity are different among groups, correlates of diversity are consistent across taxa. For both species richness and phylogenetic diversity, current precipitation and precipitation stability over time were consistently ranked among the variables that strongly correlate with diversity patterns. The correlates of phylogenetic endemism were less homogeneous across groups. The results suggest that including climate stability over time is important when predicting diversity measures that reflect historical components. Main conclusions: Investigating environmental correlates of diversity for multiple clades and diversity measures in a single geographic area allows for a better understanding of common patterns across taxa. This study shows that environmental conditions, particularly precipitation, are good predictors of the patterns of species richness and phylogenetic diversity-but not phylogenetic endemism-across different Atlantic Forest groups. Department of Biology City College of New York Ph.D. Program in Biology Graduate Center City University of New York Department of Zoology School of Biological Sciences Southern Illinois University Instituto de Geociências e Ciências Exatas UNESP-Universidade Estadual Paulista Departamento de Geografia Observatório de Dinâmicas Ecossistêmicas Instituto de Ciencias Naturales Universidad Nacional de Colombia Instituto de Biociências Universidade de São Paulo Departamento de Ecologia e Biologia Evolutiva Universidade Federal de São Paulo Departamento de Biologia Vegetal Instituto de Biologia Universidade Estadual de Campinas Universidade Federal de Santa Catarina Departamento de Biodiversidade & Centro de Aquicultura (CAUNESP) Instituto de Biociências Universidade Estadual Paulista Institute of Energy and Environment Universidade de São Paulo Departamento de Botânica Universidade Federal do Rio Grande do Sul Universidade Federal do ABC Santo André UFABC Departamento de Biologia Animal & Museu da Biodiversidade Instituto de Biologia Unicamp Universidade Federal do Paraná Institute of Systematic Botany The New York Botanical Garden Departamento de Genética e Biologia Evolutiva Instituto de Biociências Universidade de São Paulo Biological and Environmental Sciences Faculty of Natural Sciences University of Stirling Instituto de Geociências e Ciências Exatas UNESP-Universidade Estadual Paulista Departamento de Geografia Observatório de Dinâmicas Ecossistêmicas Departamento de Biodiversidade & Centro de Aquicultura (CAUNESP) Instituto de Biociências Universidade Estadual Paulista

The interdisciplinary field of Chemical Ecology in Brazil is currently composed of groups that emerged through the pioneering studies of Keith Spalding Brown Jr. and Jose Tercio Barbosa Ferreira. Following Keith Brown ‘s steps, Jose Roberto Trigo continued investigating the role of plant natural products in mediating the association among insects and their host plants, mainly in the Order Lepidoptera. The role of pyrrolizidine alkaloids in those associations was investigated extensively by Brown and Trigo, and most of what is currently known on this subject is based on their studies. The present work acknowledges their contribution to the Brazilian chemical ecology field and on insect–plant communication studies mediated by different chemical compounds.

The latitudinal diversity gradient (LDG) is arguably one of the most striking patterns in nature. The global increase in species richness toward the tropics across continents and taxonomic groups stimulated the formulation of many hypotheses to explain the underlying mechanisms of this pattern. We evaluated several of these hypotheses to explain spatial diversity patterns in the butterfly family, Nymphalidae, by assessing the contributions of speciation, extinction, and dispersal to the LDG, and also the extent to which these processes differ among regions at the same latitude. We generated a new, time-calibrated phylogeny of Nymphalidae based on 10 gene fragments and containing ca. 2,800 species (∼45% of extant diversity). Neither speciation nor extinction rate variations consistently explain the LDG among regions because temporal diversification dynamics differ greatly across longitude. For example, we found that Neotropical nymphalid diversity results from low extinction rates, not high speciation rates, and that biotic interchanges with other regions were rare. Southeast Asia was also characterized by a low speciation rate but, unlike the Neotropics, was the main source of dispersal events through time. Our results suggest that global climate change throughout the Cenozoic, particularly during the Eocene-Oligocene transition, combined with the conserved ancestral tropical niches, played a major role in generating the modern LDG of butterflies.

The present paper describes Heliconius hermathena curua Freitas & Ramos ssp. nov. This subspecies exhibits a non-mimetic phenotype typical of H. hermathena, but is characterized by the merging of the yellow streak over the forewing cubitus with the red postmedian band in the dorsal forewing. The subspecies is known from two localities in the south of Altamira, Para State, Brazil, where it inhabits an isolated patch of “campina” vegetation more than 600 km from the nearest known H. hermathena populations. Geographic isolation of the population is supported by molecular data; based on the mitochondrial gene COI, all individuals of H. hermathena curuassp. nov. form a monophyletic group and all haplotypes found in it are unique, suggesting that gene flow is not currently on-going. Given the fragile situation of Amazonian white sand forests and the proximity of the population to areas of intensive agriculture, this new subspecies and its habitat deserve attention.

The Brazilian Atlantic Rain Forest biome has been strongly impacted by human action and by various factors related to economic development. The hawkmoth Adhemarius eurysthenes (Sphingidae, Smerinthinae, Ambulycini) is endemic to the southern portion of this biome, with most of its existing populations within Brazil. Using CAD, wingless, and the barcode region of the mitochondrial COI gene, populations of A. eurysthenes were analyzed phylogeographically to obtain information on their genetic structure and variability, and to explain their spatial distribution. We obtained 14 COI haplotypes of 109 individuals from 10 localities. The population in the state of Espirito Santo, southeastern Brazil, showed the highest average genetic distance in relation to other populations. A test of molecular variance using the spatial component (SAMOVA) separated the samples into two groups; again, Espirito Santo was the divergent site. We found no significant correlation between genetic and geographic distances, but did find a tendency toward genetic isolation near the northern limit of the range of A. eurysthenes. We discuss the implementation of the observed patterns for other taxa, in an attempt to support studies on the conservation of Ambulycini in the Atlantic Rain Forest. Espirito Santo proved to be a locality of particular importance for conservation of the endemic Ambulycini in this biome.

Background Our main purpose was to evaluate the expression of plastic and evolved genes involved in ecological speciation in the noctuid moth Spodoptera frugiperda, the fall armyworm (FAW); and to demonstrate how host plants might influence lineage differentiation in this polyphagous insect. FAW is an important pest of several crops worldwide, and it is differentiated into host plant-related strains, corn (CS) and rice strains (RS). RNA-Seq and transcriptome characterization were applied to evaluate unbiased genetic expression differences in larvae from the two strains, fed on primary (corn) and alternative (rice) host plants. We consider that genes that are differently regulated by the same FAW strain, as a response to different hosts, are “plastic”. Otherwise, differences in gene expression between the two strains fed on the same host are considered constitutive differences. Results Individual performance parameters (larval and pupal weight) varied among conditions (strains vs. hosts). A total of 3657 contigs was related to plastic response, and 2395 contigs were differentially regulated in the two strains feeding on preferential and alternative hosts (constitutive contigs). Three molecular functions were present in all comparisons, both down- and up-regulated: oxidoreductase activity, metal-ion binding, and hydrolase activity. Conclusions Metabolization of foreign chemicals is among the key functions involved in the phenotypic variation of FAW strains. From an agricultural perspective, high plasticity in families of detoxifying genes indicates the capacity for a rapid response to control compounds such as insecticides.

Ticks vector diverse pathogenic bacteria that are important to identify in public health and veterinary contexts. Technological advances in high throughput sequencing have given an unprecedented opportunity to comprehensively characterize bacterial associates of ticks, but recent studies have used different 16S rRNA variable regions and sequence read lengths with little consideration of whether they reveal the same bacterial diversity. We compare the effectiveness of bacterial surveys using three library preparations across nine 16S variable regions and a set of 12 tick specimens (Acari: Ixodidae). We identify the bacterial assemblages present in extractions from wild-collected Ixodes scapularis from two regions of Canada, and provide the first microbiome survey for Ixodes angustus. Four bacterial families accounted for most diversity, with Rickettsiaceae being replaced as most common by Enterobacteriaceae or Pseudomonadaceae in some I. scapularis, and Francisellaceae being most abundant in I. angustus. The commercially available Ion 16S kit, based on 6 amplicons representing 16S regions V2, V3, V4, V67, V8 and V9, gave the most comprehensive estimates of bacterial families, with the Ion V4 amplicon generally giving the highest estimated diversity. Sequencing of the V4 amplicon by the MR DNA commercial service also provided cost effective assays of tick microbiomes that were within the range of results from the Ion 16S kit. Subtraction of the number of reads found in an extraction control sample lowered estimates of the number of bacterial families by approximately half. Our study shows that diversity patterns obtained from 16S microbiome surveys depend on the amplicon and protocol used, demonstrating that more than one marker region is needed to provide reliable inferences.

The genus Dysschema belongs to Pericopina (Erebidae: Arctiinae: Arctiini). It is outstanding because of the extreme sexual dimorphism and polymorphism found among many of its species, with males and females mimicking different groups within Lepidoptera. This has led to a confused taxonomic scenario because males and females were routinely described as different species, and an inflated number of species was established. Dysschema maginata and D. terminata are sexually dimorphic, the females of both species have the same wing pattern and represent a rare case of sex-limited mimicry. Both species were described based on the females, and the association with males was proposed long after the original descriptions without any further evidences. Despite these similarities, the former species represents a simple dual case of polymorphism, while the latter species represents an uncommon case of multiple-dual polymorphism with at least four phenotypes for the male. In this study, we tested and corroborated hypoth...

Parides ascanius (Lepidoptera: Papilionidae) is a butterfly endemic to the sand forests (“restingas”) of one of the most populated areas of Brazil (from Rio de Janeiro state to South Espirito Santo state), and was the first invertebrate officially recognized as being threatened in Brazil. Here we present a panel of eight polymorphic microsatellite loci and partial sequences of mitochondrial gene COI aiming to characterize this butterfly’s genetic diversity and understand its distribution among the extant populations. We estimate FST metrics, migration rates, cluster assignment, and spatial structure of genetic diversity. FST and statistics indicate low genetic structure and no evidence for endogamy, with all populations connected by high migration rates. Seven populations have low permanence rates (68–75 %) with increased migration probabilities for all populations. One population displays higher permanence rate (87.7 %), as the metropolitan matrix isolates it. Spatial analysis shows a global structure around the city of Rio de Janeiro and the Guanabara Bay; assignment analysis recovers six clusters evenly spread among sampled populations. These findings are consistent with a natural scenario of metapopulation dynamics for P. ascanius, with low genetic diversity and no endogamy, but progressively isolated by the metropolitan matrix. Conservation efforts should focus in connecting the isolated population, broaden the searches for new populations, and preserve all extant habitat patches where P. ascanius still occurs.

The oriental fruit moth Grapholita molesta is one of the major pests of stone and pome fruit species in Brazil. Here, we applied 1226 SNPs obtained by genotyping-by-sequencing to test whether host species associations or other factors such as geographic distance structured populations of this pest. Populations from the main areas of occurrence of G. molesta were sampled principally from peach and apple orchards. Three main clusters were recovered by neighbor-joining analysis, all defined by geographic proximity between sampling localities. Overall genetic structure inferred by a nonhierarchical amova resulted in a significant ΦST value = 0.19109. Here, we demonstrate for the first time that SNPs gathered by genotyping-by-sequencing can be used to infer genetic structure of a pest insect in Brazil; moreover, our results indicate that those markers are very informative even over a restricted geographic scale. We also demonstrate that host plant association has little effect on genetic structure among Brazilian populations of G. molesta; on the other hand, reduced gene flow promoted by geographic isolation has a stronger impact on population differentiation.

The sugarcane borer, Diatraea saccharalis (F.), is the main pest of sugarcane in Brazil. Genetic variability and gene flow among 13 Brazilian populations of the species were evaluated based on mitochondrial DNA sequences to estimate the exchange of genetic information within and among populations. We found high genetic structure among sampled localities (ΦST=0.50923), and pairwise genetic distances were significantly correlated to geographic distances. Demographic analysis and genealogical network of mitochondrial sequences indicate population growth and admixture of D. saccharalis populations, events likely related to the sequential expansion of the corn and sugarcane crops in Brazil. The implications of these findings for pest management are discussed.

The Neotropics harbour the most diverse flora and fauna on Earth. The Andes are a major centre of diversification and source of diversity for adjacent areas in plants and vertebrates, but studies on insects remain scarce, even though they constitute the largest fraction of terrestrial biodiversity. Here, we combine molecular and morphological characters to generate a dated phylogeny of the butterfly genus Pteronymia (Nymphalidae: Danainae), which we use to infer spatial, elevational and temporal diversification patterns. We first propose six taxonomic changes that raise the generic species total to 53, making Pteronymia the most diverse genus of the tribe Ithomiini. Our biogeographic reconstruction shows that Pteronymia originated in the Northern Andes, where it diversified extensively. Some lineages colonized lowlands and adjacent montane areas, but diversification in those areas remained scarce. The recent colonization of lowland areas was reflected by an increase in the rate of evolution of species’ elevational ranges towards present. By contrast, speciation rate decelerated with time, with no extinction. The geological history of the Andes and adjacent regions have likely contributed to Pteronymia diversification by providing compartmentalized habitats and an array of biotic and abiotic conditions, and by limiting dispersal between some areas while promoting interchange across others.

Population genetic studies are essential to the better application of pest management strategies, including the monitoring of the evolution of resistance to insecticides and genetically modified plants. Bacillus thuringiensis Berliner (Bt) crops have been instrumental in controlling tobacco budworm, Heliothis virescens (F.) (Lepidoptera: Noctuidae), a pest that has developed resistance to many common insecticides once used for its management. In our study, microsatellite markers were applied to investigate the genetic structure and patterns of gene flow among Brazilian populations of H. virescens from cotton, Gossypium hirsutum L., and soybean, Glycine max (L.) Merr., fields, aiming to propose means to improve its management in the field. In total, 127 alleles were found across nine microsatellites loci for 205 individuals from 12 localities. Low levels of gene flow and moderate to great genetic structure were found for these populations. Host plant association, crop growing season, and geographic origin were not responsible for the genetic structuring among Brazilian populations of H. virescens. Other factors, such as demographic history and seasonal variability of intrapopulation genetic variation, were suggested to be molding the current pattern of genetic variability distribution.

Telchin licus, the giant sugarcane borer, is an important pest species of sugarcane in northeast Brazil. Four subspecies ofTelchin licusare recognized in Brazil based on their geographic distribution and subtle differences in wing colour pattern. Some taxa are morphologically indistinguishable, and their accurate identification is key to their efficient control. Mitochondrial genes sequences (cytochrome oxidaseI and subunit 6 of thenicotinamide adenine dinucleotide dehydrogenase) were applied to delimit taxonomic entities ofT. licus, and to infer the origin of a newly established population in the state of São Paulo. The molecular data indicated that specimens sampled at different regions in Brazil are morphologically cryptic but genetically isolated entities, and at least three subspecies were assigned to the sampled localities. These data also suggested that the population collected from the state of São Paulo must have a common origin with populations from northeast Brazil, which corroborate the hypothesis that ornamental plants infested with larvae ofT. licusmight have been transported from the northeast to the southeast regions.

Intra- and inter-population genetic variability and the demographic history ofHeliothis virescens(F.) populations were evaluated by using mtDNA markers (coxI, coxIIandnad6) with samples from the major cotton- and soybean-producing regions in Brazil in the growing seasons 2007/08, 2008/09 and 2009/10. AMOVA indicated low and non-significant genetic structure, regardless of geographical scale, growing season or crop, with most of genetic variation occurring within populations. Clustering analyzes also indicated low genetic differentiation. The haplotype network obtained with combined datasets resulted in 35 haplotypes, with 28 exclusive occurrences, four of them sampled only from soybean fields. The minimum spanning network showed star-shaped structures typical of populations that underwent a recent demographic expansion. The recent expansion was supported by other demographic analyzes, such as the Bayesian skyline plot, the unimodal distribution of paired differences among mitochondrial sequences, and negative and significant values of neutrality tests for the Tajima'sDand Fu'sFSparameters. In addition, high values of haplotype diversity (Ĥ) and low values of nucleotide diversity (π), combined with a high number of low frequency haplotypes and values of θπW, suggested a recent demographic expansion ofH. virescenspopulations in Brazil. This demographic event could be responsible for the low genetic structure currently found; however, haplotypes present uniquely at the same geographic regions and from one specific host plant suggest an initial differentiation amongH. virescenspopulations within Brazil.

Polymorphic genetic markers were identified and characterized using a partial genomic library of the tobacco budworm, Heliothis virescens (Fabricius), enriched for simple sequence repeats (SSR) and nucleotide sequences of expressed sequence tags (EST). Nucleotide sequences of 192 clones from the partial genomic library yielded 147 unique SSRs while EST evaluations identified 280 SSR-containing sequences. All anonymous SSRs and 192 EST-SSRs were evaluated to select loci that produced the best quality peaks without stutter peaks. Six anonymous markers and nine EST-derived markers were selected to evaluate a sample of 96 insects collected from Stoneville, MS. The observed number of alleles ranged from two to eight, with an average of 5.87 (SE ± 0.53). The polymorphic information content (PIC) value averaged 0.446 (SE ± 0.052). Significant deviations from Hardy-Weinberg expectations were detected at eight loci. We speculate that these loci are under selection in the population of tobacco budworm use...

Global biodiversity peaks in the tropical forests of the Andes, a striking geological feature that has likely been instrumental in generating biodiversity by providing opportunities for both vicariant and ecological speciation. However, the role of these mountains in the diversification of insects, which dominate biodiversity, has been poorly explored using phylogenetic methods. Here we study the role of the Andes in the evolution of a diverse Neotropical insect group, the clearwing butterflies. We used dated species-level phylogenies to investigate the time course of speciation and to infer ancestral elevation ranges for two diverse genera. We show that both genera likely originated at middle elevations in the Andes in the Middle Miocene, contrasting with most published results in vertebrates that point to a lowland origin. Although we detected a signature of vicariance caused by the uplift of the Andes at the Miocene-Pliocene boundary, most sister species were parapatric without any obvious vicariant barrier. Combined with an overall decelerating speciation rate, these results suggest an important role for ecological speciation and adaptive radiation, rather than simple vicariance.

A new species of Actinote Hubner, A. kennethi Freitas, Willmott and Hall (Lepidoptera: Nymphalidae: Heliconiinae), is described from cloud forest habitats in eastern Ecuador. Molecular sequence data and morphological characters both indicate that the new taxon is closely related to Actinote g. genitrix d'Almeida, 1922, from southeastern Brazil, and the Venezuelan Actinote genitrix costae Neild, 2008. Based on the substantial geographic isolation of these taxa, observed morphological differences and mtDNA sequence divergence, we argue for the treatment of the new Ecuadorian Actinote taxon as a distinct species.

The tribe Acraeini (Nymphalidae, Heliconiinae) is believed to comprise between one and seven genera, with the greatest diversity in Africa. The genera Abananote, Altinote, and Actinote (s. str.) are distributed in the Neotropics, while the genera Acraea, Bematistes, Miyana, and Pardopsis have a Palaeotropical distribution. The monotypic Pardopsis use herbaceous plants of the family Violaceae, Acraea and Bematistes feed selectively on plants with cyanoglycosides belonging to many plant families, but preferentially to Passifloraceae, and all Neotropical species with a known life cycle feed on Asteraceae only. Here, a molecular phylogeny is proposed for the butterflies of the tribe Acraeini based on sequences of COI, EF-1alpha and wgl. Both Maximum Parsimony and Bayesian analyses showed that the tribe is monophyletic, once the genus Pardopsis is excluded, since it appears to be related to Argynnini. The existing genus Acraea is a paraphyletic group with regard to the South American genera, and the species of Acraea belonging to the group of "Old World Actinote" is the sister group of the Neotropical genera. The monophyly of South American clade is strongly supported, suggesting a single colonization event of South America. The New World Actinote (s. str.) is monophyletic, and sister to Abananote+Altinote (polyphyletic). Based on the present results it was possible to propose a scenario for the evolution in host plant use within Acraeini, mainly concerning the use of Asteraceae by the South American genera.

Molecular phylogenetic analyses were conducted to determine relationships and to investigate character evolution for the Troidini/Aristolochia interaction, in an attempt to answer the following questions: (1) what is the present pattern of use of Aristolochia by these butterflies; (2) is the pattern we see today related to the phylogeny of plants or to their chemical composition; (3) can the geographical distribution of Aristolochia explain the host plant use observed today; and (4) how did the interaction between Troidini and Aristolochia evolve? Analyses of character optimization suggest that the current pattern of host plant use of these butterflies does not seem to be constrained by the phylogeny of their food plants, neither by the secondary chemicals in these plants nor by their geographical similarity. The current host plant use in these butterflies seems to be simply opportunistic, with species with a wider geographical range using more species of host plants than those with a more restricted distribution.

The recent publication of a time-tree for the plant family Solanaceae (nightshades) provides the opportunity to use independent calibrations to test divergence times previously inferred for the diverse Neotropical butterfly tribe Ithomiini. Ithomiini includes clades that are obligate herbivores of Solanaceae, with some genera feeding on only one genus. We used 8 calibrations extracted from the plant tree in a new relaxed molecular-clock analysis to produce an alternative temporal framework for the diversification of ithomiines. We compared the resulting age estimates to: (i) a time-tree obtained using 7 secondary calibrations from the Nymphalidae tree of Wahlberg et al. (2009), and (ii) Wahlberg et al.'s (2009) original age estimates for the same clades. We found that Bayesian clock estimates were rather sensitive to a variety of analytical parameters, including taxon sampling. Regardless of this sensitivity however, ithomiine divergence times calibrated with the ages of nightshades were always on average half the age of previous estimates. Younger dates for ithomiine clades appear to fit better with factors long suggested to have promoted diversification of the group such as the uplifting of the Andes, in the case of montane genera. Alternatively, if ithomiines are as old as previous estimates suggest, the recent ages inferred for the diversification of Solanaceae seem likely to be seriously underestimated. Our study exemplifies the difficulty of testing hypotheses of divergence times and of choosing between alternative dating scenarios, and shows that age estimates based on seemingly plausible calibrations may be grossly incongruent.

In this report, a molecular phylogeny based on the plastid gene matK and on the region between the genes trnL and trnF, and a chemical relationship based on the pattern of sesquiterpenes, is proposed for Aristolochia species from southeastern Brazil. We found that Aristolochia is a monophyletic genus and species considered to be derived contain labdanoic acids (LAs) in their leaves. The phenetic relationship recovered with sesquiterpenes did not agree with the phylogenetic relationships for Aristolochia, and three main clusters were recognized, namely, germacrene-D, germacrene-C and Z-caryophyllene groups. The presence of different sesquiterpene structures in species that are phylogenetically closely related may reflect adaptations to avoid predation of herbivores specialized in feeding on Aristolochiaceae plants.

A phylogenetic hypothesis for the tribe Ithomiini (Lepidoptera: Nymphalidae: Danainae) is presented, based on sequences of the mitochondrial cytochrome oxidase subunits I and II (COI-COII) region and regions of the nuclear genes wingless and Elongation factor 1-alpha. Branch support for each clade is assessed, and a partition congruence index is used to explore conflict among gene regions. The monophyly of the clade is strongly supported, as are many of the traditionally recognized subtribes and genera. The data imply para- phyly of some genera and tribes, but largely support recent classifications and phylogenetic hypotheses based on morphological characters.

Proteins containing an Rho GTPase-activating protein (RhoGAP) domain work as molecular switches involved in the regulation of diverse cellular functions. The ability of these GTPases to regulate a wide number of cellular processes conies from their interactions with multiple effectors and inhibitors, including the RhoGAP family, which stimulates their intrinsic GTPase activity. Here, a phylogenetic approach was applied to study the evolutionary relationship among 59 RhoGAP domain-containing proteins. The sequences were aligned by their RhoGAP domains and the phylogenetic hypotheses were generated using Maximum Parsimony and Bayesian analyses. The character tracing of two traits, GTPase activity and presence of other domains, indicated a significant phylogenetic signal for both of them.

The pantropical weed Bidens pilosa (Asteraceae) is a species with several taxonomic problems. Recently it has been shown to be a complex of different species. To shed light on this problem in Brazil, dichloromethane extract of leaves of several populations corresponding to three proposed species for the complex in southeastern Brazil (B. pilosa, Bidens alba and Bidens subalternans) were analyzed by GC–MS. Twenty-four substances were detected, of which four resemble polyacetylenes, the others sesquiterpenes. Five sesquiterpenes tentatively identified as E-caryophyllene, α-humulene, germacrene-D, bicyclogermacrene and α-muurolene were found in all three Bidens species. The polyacetylene phenylhepta-1,3,5-triyne was identified only in B. alba. Multivariate analysis (cluster and principal component analyses) separated the three entities, suggesting that these compounds could represent a useful tool to distinguish species in the B. pilosa complex.

Morphological and molecular data from two prior publications are combined with new data resulting in a mostly well-supported cladistic hypothesis of relationships among subtribes and genera of Ithomiini, a diverse clade of Neotropical butterflies. A revised classification for relationships among the nine tribes and 46 genera comprising the group is proposed, and Athesitina new subtribe is described for Athesis Doubleday, 1847 and Patricia Fox, 1940. Oxapampa Brabant, 2004 is synonymized with Veladyris Fox, 1945. http://zoobank.org/urn:lsid:zoobank.org:pub:3EE6B75B-94B3-42FF-8149-1E9A18417438

The genus Hermeuptychia is common and widespread through the Americas, from Argentina to the southern United States of America. All eight recognized species within Hermeuptychia are small and brown, with very similar interspecific external morphologies and intraspecifically variable ocelli patterns that render taxonomic identification based on morphology difficult. In our study, we surveyed variability within Hermeuptychia, and evaluated species boundaries based on molecular data (sequences of the ‘barcode’ mitochondrial DNA COI gene) and morphology (mainly male genitalia), using a phylogenetic approach. We found eight DNA-based and 12 morphological groups in our sampling. Species names were assigned based mainly on comparisons with male genitalia morphology descriptions corresponding to name-bearing type specimens. Morphological and DNA variability were highly congruent, with the exception of group H, the Hermeuptychia cucullina complex. Also, the barcode region showed a clear threshold for intra- and interspecific mean distances around 2%. Based on these results, we circumscribe the species boundaries in the genus Hermeuptychia and discuss conflicts between mitochondrial genes and classic morphological approaches for identifying and delimiting species. Our study revealed cryptic diversity within an ubiquitous genus of Neotropical butterflies.

Population genetic studies are essential to the better application of pest management strategies, including the monitoring of the evolution of resistance to insecticides and genetically modified plants. Bacillus thuringiensis Berliner (Bt) crops have been instrumental in controlling tobacco budworm, Heliothis virescens (F.) (Lepidoptera: Noctuidae), a pest that has developed resistance to many common insecticides once used for its management. In our study, microsatellite markers were applied to investigate the genetic structure and patterns of gene flow among Brazilian populations of H. virescens from cotton, Gossypium hirsutum L., and soybean, Glycine max (L.) Merr., fields, aiming to propose means to improve its management in the field. In total, 127 alleles were found across nine microsatellites loci for 205 individuals from 12 localities. Low levels of gene flow and moderate to great genetic structure were found for these populations. Host plant association, crop growing season, and geographic origin were not responsible for the genetic structuring among Brazilian populations of H. virescens. Other factors, such as demographic history and seasonal variability of intrapopulation genetic variation, were suggested to be molding the current pattern of genetic variability distribution.

Background The temporal and geographical diversification of Neotropical insects remains poorly understood because of the complex changes in geological and climatic conditions that occurred during the Cenozoic. To better understand extant patterns in Neotropical biodiversity, we investigated the evolutionary history of three Neotropical swallowtail Troidini genera (Papilionidae). First, DNA-based species delimitation analyses were conducted to assess species boundaries within Neotropical Troidini using an enlarged fragment of the standard barcode gene. Molecularly delineated species were then used to infer a time-calibrated species-level phylogeny based on a three-gene dataset and Bayesian dating analyses. The corresponding chronogram was used to explore their temporal and geographical diversification through distinct likelihood-based methods. Results The phylogeny for Neotropical Troidini was well resolved and strongly supported. Molecular dating and biogeographic analyses indicate that the extant lineages of Neotropical Troidini have a late Eocene (33–42 Ma) origin in North America. Two independent lineages (Battus and Euryades + Parides) reached South America via the GAARlandia temporary connection, and later became extinct in North America. They only began substantive diversification during the early Miocene in Amazonia. Macroevolutionary analysis supports the “museum model” of diversification, rather than Pleistocene refugia, as the best explanation for the diversification of these lineages. Conclusions This study demonstrates that: (i) current Neotropical biodiversity may have originated ex situ; (ii) the GAARlandia bridge was important in facilitating invasions of South America; (iii) colonization of Amazonia initiated the crown diversification of these swallowtails; and (iv) Amazonia is not only a species-rich region but also acted as a sanctuary for the dynamics of this diversity. In particular, Amazonia probably allowed the persistence of old lineages and contributed to the steady accumulation of diversity over time with constant net diversification rates, a result that contrasts with previous studies on other South American butterflies.

Parides burchellanus is considered a rare and threatened swallowtail species restricted to central Brazil. It shows considerable morphological similarity to Parides panthonus jaguarae, with which it shares both geographical range and larval host plant. At present, P. burchellanus and P. panthonus are believed to be distinct species, based on minor differences in male genitalia. In this study, the phylogenetic and systematic position of P. burchellanus in relation to three subspecies of P. panthonus (P. p. jaguarae, P. p. lysimachus and P. p. aglaope) was evaluated using molecular evidence: the complete sequence of the mtDNA genes COI and COII and of the nuclear gene EF-1α (c. 3300 bp). In addition, the informativeness of the 'barcode' region next to the 5' end of COI (c. 650 bp) was evaluated for delimiting these taxa. Individual analysis by neighbour-joining, using Kimura 2-parameter distance model, and by maximum parsimony showed that P. p. jaguarae + P. p. lysimachus + P. p. aglaope + P. burchellanus form a strongly supported monophyletic clade, and all molecular regions consistently recovered P. p. jaguarae and P. burchellanus as sister species. The genetic divergence among the subspecies of P. panthonus and P. burchellanus is equivalent to the divergence among conspecifics of other species of Parides, and smaller than the interspecific divergence among different sister species of this genus. The results support the proposal that P. p. jaguarae and P. burchellanus are likely to be synonymous, and suggest that P. burchellanus can be considered conspecific with P. panthonus. The taxonomic classification of P. burchellanus should be revised on the basis of the molecular data.

Faced by a growing need of identification and delimitation of new and established cryptic species that are being lost at an increasing rate, taxonomists can now more than ever take advantage of an enormous variety of new molecular and computational tools. At this moment they should be open to all new available technologies in the so called "technology-driven revolution" in systematics. The use of the "DNA barcode" has been discussed by those applying successfully this approach to identify and diagnose species and by those who believe that the flaws in the use of this molecular marker are as many as to negate the worth of its employment. For insects of the order Lepidoptera neither side seems totally correct orwrong, and although many groups of lepidopterans have been taxonomically resolved by using exclusively or additionally this marker for diagnoses, for others the "barcode" helped little to resolve taxonomic issues. Here we briefly present some pros and cons of using DNA barcode as a tool in taxonomic studies, with special attention to studies with groups of Lepidoptera developed in the last few years. A necessida de crescente de identificação e delimitação de novas espécies, ou de espécies crípticas já estabelecidas, que estão sendo perdidas a uma taxa também crescente, tem levado diversos especialistas a utilizar em uma variedade de ferramentas moleculares e computacionais. Neste momento, taxonomistas devem estar atentos a toda nova tecnologia disponível na chamada "revolução dirigida pela tecnologia"na sistemática, que tem entre as novas ferramentas moleculares a utilização de"DNA barcodes". O uso de "DNA barcode" tem sido amplamente discutido por aqueles que aplicam essa abordagem com sucesso para identificar e diagnosticar espécies, e por aqueles que acreditam que são tantos os problemas no uso desse marcador molecular que não se justifica seu emprego. Para insetos da ordem Lepidoptera nenhum lado parece estar totalmente certo ou errado e, embora alguns grupos de lepidópteros tenham sido resolvidos taxonomicamente pelo uso exclusivo ou adicional desse marcador, para outros o "barcode" ajudou pouco a resolver problemas taxonômicos. Aqui nós apresentamos brevemente prós e contras do uso de "DNA barcode" como ferramenta em estudos taxonômicos, com atenção especial para estudos com grupos de Lepidoptera desenvolvidos nos últimos anos.

Hypocrealean arthropod pathogenic fungi have profound impact on the regulation of agricultural and medical pests. However, until now the genetic and phylogenetic relationships among species have not been clarified, such studies could clarify host specificity relationships and define species boundaries. Our purpose was to compare the sequences of the mitochondrial SSU rDNA fragments from several mitosporic entomopathogenic Hypocreales to infer relationships among them and to evaluate the possibility to use these sequences as species diagnostic tool in addition to the more commonly studied sequences of nuclear SSU rDNA. The SSU mt-rDNA proved to be useful to help in differentiation of species inside several genera. Clusters obtained with Parsimony, Bayesian, and Maximum Likelihood analyses were congruent with a new classification of the Clavicipitaceae (Sung et al. Stud Mycol. 2007;57:5-59) in which the anamorphic genera Nomuraea and Metarhizium species remain in the Clavicipitaceae and Isaria species sequenced here are assigned to the family Cordycipitaceae. Mitochondrial genomic information indicates the same general pattern of relationships demonstrated by nuclear gene sequences.

A phylogeny of the Neotropical members of the Tribe Troidini (Lepidoptera: Papilionidae) was obtained with sequences of three protein-coding genes: two mitochondrial (COI and COII), and one nuclear (EF-1alpha). Parsimony and Bayesian analyses of 33 taxa resulted in very similar trees regardless of method used with the 27 troidines always forming a monophyletic clade. Within Troidini, the genus Battus is sister group to the remaining troidines, followed by a clade formed by the Paleotropical taxa (here represented by three exemplars). The genus Euryades is the next branch, and sister group of Parides. The genus Parides is monophyletic, and is divided into four main groups by Maximum Parsimony analysis, with the most basal group composed of tailed species restricted to SE Brazil. Character optimization of ecological and morphological traits over the phylogeny proposed for troidines indicated that the use of several species of Aristolochia is ancestral over the use of few or a single host-plant. For the other three characters, the ancestral states were the absence of long tails, forest as the primary habitat and oviposition solitary or in loose group of several eggs.

This paper describes a new, abundant and widespread species of Yphthimoides Forster from the Atlantic forests of southern Brazil, Paraguay, and northern Argentina (Misiones) in open and secondary vegetation and forest edges. Adult and immature stage morphology is described, molecular data are provided, and the placement of the new species within the genus Yphthimoides is discussed.