Degree Of Reproductive Isolation Research Articles

Phylogenetic insight into the Lecidea atrobrunnea complex – evidence of narrow geographic endemics and the pressing need for integrative taxonomic revisions

AbstractSpecies of lichen-forming fungi (LFF) display an array of geographical distribution patterns. Among the broadly distributed lichen-forming fungal species, the degree of reproductive isolation and genetic substructure among populations varies widely, in some cases masking unrecognized diversity or meaningful biogeographical patterns. Lecidea atrobrunnea (Raymond ex Lam. & DC.) Schaer. s. lat. (Lecideaceae) is a widespread species complex that has been studied for over two centuries since its initial description. The diversity of the L. atrobrunnea group is highest in western North America, where a dizzying array of morphologies and chemistry can occur at local scales. Here we investigate whether the assumed cosmopolitan distribution of L. atrobrunnea s. lat. is an artifact of taxonomic limitations and masks biogeographical patterns in this species complex. To address these questions, we compiled sequence data from the standard fungal barcoding marker (ITS) for over 100 specimens within this complex, in addition to genome-scale data from a subset of these representing over 1600 single-copy nuclear genes spanning over 3 Mb of the genome. Our study corroborates the perspective that the morphologically and chemically variable Lecidea atrobrunnea group reflects a complex of distinct species-level lineages, with 42–83 candidate species inferred from the ITS region and high levels of diversity inferred from a subset of specimens using genome-scale data. However, both phenotype- and molecular-based species boundaries remained unsettled, with the most common nominal taxa recovered as highly polyphyletic and with conflict among different molecular species delimitation approaches. Our study also highlights the potential for geographically restricted species, with fascinating biogeographical patterns, challenging, in part, the assumed cosmopolitan distribution of L. atrobrunnea s. lat. This study provides valuable direction for future research that will be crucial in understanding diversification and establishing a robust taxonomy for this well-known species complex.

Israeli Oncocyclus irises: Phylogenetic relationships and evolutionary history

Abstract Oncocyclus is a large group of bearded irises with poorly known phylogeny and evolutionary history. In Israel, Iris sect. Oncocyclus comprises eight species belonging to three aggregates. We used a combination of approaches to resolve the phylogenetic relationships of these species and indicate the evolutionary forces responsible for their origin. We sequenced the whole chloroplast genomes of species and integrated a phylogenetic tree with results of genetic (AFLP) divergence, degree of reproductive isolation, and species distribution modeling. Our findings suggest that quantitative and even qualitative morphological characters, such as flower color, are unreliable diagnostic traits for Oncocyclus taxonomy; that some recognized species comprise more than one species; and that group evolution did not involve the origin of distinct flower aggregates. A lack of pre-zygotic reproductive isolation agrees with the very low variability of the Oncocyclus plastome, suggesting that Israeli Oncocyclus species are very young. Homoploid hybridization followed or preceded by long periods of geographic isolation, and local selection likely contributed to speciation in Oncocyclus. In the group evolutionary history, importance of homoploid hybridization and local selection differed among species, but limited gene flow played a crucial role for all species.

Patterns of genome size evolution versus fraction of repetitive elements in statu nascendi species: the case of the willistoni subgroup of Drosophila (Diptera, Drosophilidae).

Genome size evolution is known to be related with transposable elements, yet such relation in incipient species remains poorly understood. For decades, the willistoni subgroup of Drosophila has been a model for evolutionary studies because of the different evolutionary stages and degrees of reproductive isolation its species present. Our main question here was how speciation influences genome size evolution and the fraction of repetitive elements, with a focus on transposable elements. We quantitatively compared the mobilome of four species and two subspecies belonging to this subgroup with their genome size, and performed comparative phylogenetic analyses. Our results showed that genome size and the fraction of repetitive elements evolved according to the evolutionary history of these species, but the content of transposable elements showed some discrepancies. Signals of recent transposition events were detected for different superfamilies. Their low genomic GC content suggests that in these species transposable element mobilization might be facilitated by relaxed natural selection. Additionally, a possible role of the superfamily DNA/TcMar-Tigger in the expansion of these genomes was also detected. We hypothesize that the undergoing process of speciation could be promoting the observed increase in the fraction of repetitive elements and, consequently, genome size.

Mitonuclear discordance and patterns of reproductive isolation in a complex of simultaneously hermaphroditic species, the Allolobophora chlorotica case study

Historical events of population fragmentation, expansion and admixture over geological time may result in complex patterns of reproductive isolation and may explain why, for some taxa, the study of mitochondrial (mt) and nuclear (nu) genetic data results in discordant evolutionary patterns. Complex patterns of taxonomic diversity were recently revealed in earthworms for which distribution is largely the result of paleogeographical events. Here, we investigated reproductive isolation patterns in a complex of cryptic species of earthworms in which discordant patterns between mt and nu genetic lineages were previously revealed, the Allolobophora chlorotica aggregate. Using four nu microsatellite markers and a fragment of the cytochrome c oxidase subunit I mt gene, we carried out a parentage analysis to investigate the mating patterns (i) between individuals belonging to two divergent mt lineages that cannot be distinguished with nu markers and (ii) between individuals belonging to lineages that are differentiated both at the mt and nu levels. Amongst the 157 field‐collected individuals, 66 adults were used in cross‐breeding experiments to form 22 trios based on their assignment to a mt lineage, and 453 obtained juveniles were genotyped. We showed that adults that mated with both their potential mates in the trio produced significantly more juveniles. In crosses between lineages that diverged exclusively at the mt level, a sex‐specific pattern of reproduction characteristic to each lineage was observed, suggesting a possible conflict of interest concerning the use of male/female function between mating partners. In crosses between lineages that diverged both at the mt and nu levels, a high production of cocoons was counterbalanced by a low hatching rate, suggesting a post‐zygotic reproductive isolation. Different degrees of reproductive isolation, from differential sex allocation to post‐zygotic isolation, were thus revealed. Lineages appear to be at different stages in the speciation process, which likely explain the observed opposite patterns of mitonuclear congruence.

Contemporary Philosophy and Methods for Studying Speciation and Delimiting Species

Species delimitation is a first step for realizing the extent of biodiversity and is relevant for all downstream applications in biology. The production of large genome-scale datasets for non-model organisms combined with the development of methodological tools have allowed researchers to examine fine-scale processes of speciation such as timing of origin, degree of migration, population-size changes, selection, drift, and recombination. Studies using reptiles and amphibians have, in part, paved the way for the development and use of such methods for exploring speciation and delimitation. While these methodologies have improved our understanding of processes of diversification, researchers are far from agreeing upon a set of criteria to delimit species. In cases where genetic lineages are discovered that are unique to geographic areas, researchers usually agree that two entities exist. Disagreement about taxonomic status often centers on the degree of reproductive isolation between taxa and probability of remaining distinct. However, reproductive isolation is frequently inferred without examining gene flow, understanding the nature of hybrid zones, or determining the amount and type of introgression. Here, we review some of the vexing problems for delimiting reptiles and amphibians, which include isolation by distance, gene flow and differential allelic introgression, hybrid zone dynamics, and the nature of genomic islands of divergence. We also respond to recent literature criticizing model-based species delimitation in North American snakes in the context of these methodological advancements and address how scientists can move forward with studies on speciation.

Patterns of reproductive isolation in a haplodiploid mite, Amphitetranychus viennensis: prezygotic isolation, hybrid inviability and hybrid sterility

BackgroundEvolution of reproductive isolation is an important process, generating biodiversity and driving speciation. To better understand this process, it is necessary to investigate factors underlying reproductive isolation through various approaches but also in various taxa. Previous studies, mainly focusing on diploid animals, supported the prevalent view that reproductive barriers evolve gradually as a by-product of genetic changes accumulated by natural selection by showing a positive relationship between the degree of reproductive isolation and genetic distance. Haplodiploid animals are expected to generate additional insight into speciation, but few studies investigated the prevalent view in haplodiploid animals. In this study, we investigate whether the relationship also holds in a haplodiploid spider mite, Amphitetranychus viennensis (Zacher).ResultsWe sampled seven populations of the mite in the Palaearctic region, measured their genetic distance (mtDNA) and carried out cross experiments with all combinations. We analyzed how lack of fertilization rate (as measure of prezygotic isolation) as well as hybrid inviability and hybrid sterility (as measures of postzygotic isolation) varies with genetic distance. We found that the degree of reproductive isolation varies among cross combinations, and that all three measures of reproductive isolation have a positive relationship with genetic distance. Based on the mtDNA marker, lack of fertilization rate, hybrid female inviability and hybrid female sterility were estimated to be nearly complete (99.0–99.9% barrier) at genetic distances of 0.475–0.657, 0.150–0.209 and 0.145–0.210, respectively. Besides, we found asymmetries in reproductive isolation.ConclusionsThe prevalent view on the evolution of reproductive barriers is supported in the haplodiploid spider mite we studied here. According to the estimated minimum genetic distance for total reproductive isolation in parent population crosses in this study and previous work, a genetic distance of 0.15–0.21 in mtDNA (COI) appears required for speciation in spider mites. Variations and asymmetries in the degree of reproductive isolation highlight the importance of reinforcement of prezygotic reproductive isolation through incompatibility and the importance of cytonuclear interactions for reproductive isolation in haplodiploid spider mites.

Limited genetic parallelism underlies recent, repeated incipient speciation in geographically proximate populations of an Arctic fish (Salvelinus alpinus).

The genetic underpinnings of incipient speciation, including the genomic mechanisms which contribute to morphological and ecological differentiation and reproductive isolation, remain poorly understood. The repeated evolution of consistently, phenotypically distinct morphs of Arctic Charr (Salvelinus alpinus) within the Quaternary period offer an ideal model to study the repeatability of evolution at the genomic level. Sympatric morphs of Arctic Charr are found across this species' circumpolar distribution. However, the specific genetic mechanisms driving this morph differentiation are largely unknown despite the cultural and economic importance of the anadromous morph. We used a newly designed 87k SNP chip to investigate the character and consistency of the genomic differences among sympatric morphs within three recently deglaciated and geographically proximate lakes in Labrador, Canada. We found genetically distinct small and large morph Arctic Charr in all three lakes consistent with resident and anadromous morphs, respectively. A degree of reproductive isolation among sympatric morphs is likely given genome-wide distributions of outlier SNPs and high genome-wide FST s. Across all lakes, outlier SNPs were largely nonoverlapping suggesting a lack of genetic parallelism driving morph differentiation. Alternatively, several genes and paralogous copies of the same gene consistently differentiated morphs across multiple lakes suggesting their importance to the manifestation of morphs. Our results confirm the utility of Arctic Charr as a model for investigating the predictability of evolution and support the importance of both genetic parallelism and nonparallelism to the incipient speciation of Arctic Charr morphs.

Speciation through chromosomal fusion and fission in Lepidoptera.

Changes in chromosome numbers may strongly affect reproductive barriers, because individuals heterozygous for distinct karyotypes are typically expected to be at least partially sterile or to show reduced recombination. Therefore, several classic speciation models are based on chromosomal changes. One import mechanism generating variation in chromosome numbers is fusion and fission of existing chromosomes, which is particularly likely in species with holocentric chromosomes, i.e. chromosomes that lack a single centromere. Holocentric chromosomes evolved repeatedly across the tree of life, including in Lepidoptera. Although changes in chromosome numbers are hypothesized to be an important driver of the spectacular diversification of Lepidoptera, comparative studies across the order are lacking. We performed the first comprehensive literature survey of karyotypes for Lepidoptera species since the 1970s and tested if, and how, chromosomal variation might affect speciation. Even though a meta-analysis of karyological differences between closely related taxa did not reveal an effect on the degree of reproductive isolation, phylogenetic diversification rate analyses across the 16 best-covered genera indicated a strong, positive association of rates of chromosome number evolution and speciation. These findings suggest a macroevolutionary impact of varying chromosome numbers in Lepidoptera and likely apply to other taxonomic groups, especially to those with holocentric chromosomes. This article is part of the theme issue 'Towards the completion of speciation: the evolution of reproductive isolation beyond the first barriers'.

The Mendelian inheritance of gynomonoecy: insights from Anacyclus hybridizing species.

Gynomonoecy is an infrequent sexual system in angiosperms, although widely represented within the Asteraceae family. Currently, the hypothesis of two nuclear loci controling gynomonoecy is the most accepted. However, the genic interactions are still uncertain. Anacyclus clavatus, A. homogamos, and A. valentinus differ in their sexual system and floral traits. Here, we investigate the inheritance of gynomonoecy in this model system to understand its prevalence in the family. We selected six natural populations (two per species) for intra- and interspecific experimental crosses, and generated a total of 1123 individuals from the F1 generation, F2 , and backcrosses for sexual system characterization. The frequency of gynomonoecy observed for each cross was tested to fit different possible hypotheses of genic interaction. Additionally, the breeding system and the degree of reproductive isolation between these species were assessed. Complementary epistasis, in which two dominant alleles are required for trait expression, explained the frequencies of gynomonoecy observed across all generations. The heterozygosity inferred in Anacyclus valentinus, as well as its lower and variable seed set, is congruent with its hybrid origin. In our model system gynomonoecy is controlled by complementary epistasis of two genes. A common origin of this sexual system in Asteraceae, in which genic duplications, mutations, and hybridization between lineages played a key role, is hypothesized whereas independent evolutionary pathways and possibly diverse underlying genetic factors are suggested for gynomonoecy expression in other angiosperm families.

Symbionts do not affect the mating incompatibility between the Brazilian-1 and Peruvian morphotypes of the Anastrepha fraterculus cryptic species complex

The South American fruit fly, Anastrepha fraterculus, is clearly undergoing a speciation process. Among others, two of their morphotypes, the Brazilian-1 and Peruvian, have accumulated differences in pre- and post-zygotic mechanisms resulting in a degree of reproductive isolation. Both harbor a different strain of Wolbachia, which is a widespread endosymbiotic bacterium among many invertebrates producing a range of reproductive effects. In this paper, we studied the role of this bacterium as one of the factors involved in such isolation process. Infected and cured laboratory colonies were used to test pre- and post-zygotic effects, with special emphasis in uni- and bi-directional cytoplasmic incompatibility (CI). We showed that Wolbachia is the only known reproductive symbiont present in these morphotypes. Wolbachia reduced the ability for embryonic development in crosses involving cured females and infected males within each morphotype (uni-directional CI). This inhibition showed to be more effective in the Peruvian morphotype. Bi-directional CI was not evidenced, suggesting the presence of compatible Wolbachia strains. We conclude that Wolbachia is not directly involved in the speciation process of these morphotypes. Other mechanisms rather than CI should be explored in order to explain the reduced mating compatibility between the Brazilian-1 and Peruvian morphotypes.

Establishment, Hybridization, Dispersal, Impact, and Decline of Diorhabda spp. (Coleoptera: Chrysomelidae) Released for Biological Control of Tamarisk in Texas and New Mexico.

Three Diorhabda spp. tamarisk beetles (Coleoptera: Chrysomelidae) were established in Texas from 2003 to 2010 for biological control of tamarisk (Tamarix spp.): Mediterranean tamarisk beetles, D. elongata (Brullé) from Greece, also established in New Mexico; subtropical tamarisk beetles, D. sublineata (Lucas) from Tunisia; and larger tamarisk beetles, D. carinata (Faldermann) from Uzbekistan. More than one million tamarisk beetles were released at 99 sites. Species establishment success ranged from 52 to 83%. All three species now co-occur in New Mexico with the northern tamarisk beetles, D. carinulata (Desbrochers). A phenotypic hybrid scoring system was developed to assess Diorhabda phenotype distributions and character mixing in hybrid zones. Widespread field populations of bispecific hybrid phenotypes for D. carinata/D. elongata and D. sublineata/D. elongata rapidly appeared following contact of parental species. Initial distributions and dispersal of Diorhabda spp. and hybrids are mapped for Texas, New Mexico, Oklahoma, and Kansas, where they produced large-scale tamarisk defoliation and localized dieback for 3-4 yr. However, populations subsequently severely declined, now producing only isolated defoliation and allowing tamarisk to recover. Diorhabda sublineata and D. elongata temporarily produced nontarget spillover defoliation of ornamental athel, Tamarix aphylla (L.) Karst, along the Rio Grande. Hybrid phenotypes were generally bimodally distributed, indicating some degree of reproductive isolation. Additional diagnostic phenotypic characters in males allowed more precise hybrid scoring. Character mixing in some hybrid populations approached or reached that of a hybrid swarm. The significance of hybridization for tamarisk biocontrol is discussed.

Extensive genetic differentiation between recently evolved sympatric Arctic charr morphs.

The availability of diverse ecological niches can promote adaptation of trophic specializations and related traits, as has been repeatedly observed in evolutionary radiations of freshwater fish. The role of genetics, environment, and history in ecologically driven divergence and adaptation, can be studied on adaptive radiations or populations showing ecological polymorphism. Salmonids, especially the Salvelinus genus, are renowned for both phenotypic diversity and polymorphism. Arctic charr (Salvelinus alpinus) invaded Icelandic streams during the glacial retreat (about 10,000 years ago) and exhibits many instances of sympatric polymorphism. Particularly, well studied are the four morphs in Lake Þingvallavatn in Iceland. The small benthic (SB), large benthic (LB), planktivorous (PL), and piscivorous (PI) charr differ in many regards, including size, form, and life history traits. To investigate relatedness and genomic differentiation between morphs, we identified variable sites from RNA‐sequencing data from three of those morphs and verified 22 variants in population samples. The data reveal genetic differences between the morphs, with the two benthic morphs being more similar and the PL‐charr more genetically different. The markers with high differentiation map to all linkage groups, suggesting ancient and pervasive genetic separation of these three morphs. Furthermore, GO analyses suggest differences in collagen metabolism, odontogenesis, and sensory systems between PL‐charr and the benthic morphs. Genotyping in population samples from all four morphs confirms the genetic separation and indicates that the PI‐charr are less genetically distinct than the other three morphs. The genetic separation of the other three morphs indicates certain degree of reproductive isolation. The extent of gene flow between the morphs and the nature of reproductive barriers between them remain to be elucidated.

Small and genetically highly structured populations in a long-legged bee, Rediviva longimanus, as inferred by pooled RAD-seq

Adaptation to local host plants may impact a pollinator’s population genetic structure by reducing gene flow and driving population genetic differentiation, representing an early stage of ecological speciation. South African Rediviva longimanus bees exhibit elongated forelegs, a bizarre adaptation for collecting oil from floral spurs of their Diascia hosts. Furthermore, R. longimanus foreleg length (FLL) differs significantly among populations, which has been hypothesised to result from selection imposed by inter-population variation in Diascia floral spur length. Here, we used a pooled restriction site-associated DNA sequencing (pooled RAD-seq) approach to investigate the population genetic structure of R. longimanus and to test if phenotypic differences in FLL translate into increased genetic differentiation (i) between R. longimanus populations and (ii) between phenotypes across populations. We also inferred the effects of demographic processes on population genetic structure and tested for genetic markers underpinning local adaptation.ResultsPopulations showed marked genetic differentiation (average FST = 0.165), though differentiation was not statistically associated with differences between populations in FLL. All populations exhibited very low genetic diversity and were inferred to have gone through recent bottleneck events, suggesting extremely low effective population sizes. Genetic differentiation between samples pooled by leg length (short versus long) rather than by population of origin was even higher (FST = 0.260) than between populations, suggesting reduced interbreeding between long and short-legged individuals. Signatures of selection were detected in 1119 (3.8%) of a total of 29,721 SNP markers,ConclusionsPopulations of R. longimanus appear to be small, bottlenecked and isolated. Though we could not detect the effect of local adaptation (FLL in response to floral spurs of host plants) on population genetic differentiation, short and long legged bees appeared to be partially differentiated, suggesting incipient ecological speciation. To test this hypothesis, greater resolution through the use of individual-based whole-genome analyses is now needed to quantify the degree of reproductive isolation between long and short legged bees between and even within populations.

First record of Culex (Culex) bidens (Diptera: Culicidae) in Colombia: Taxonomic and epidemiological implications

Arbovirus transmission cycles must be studied locally since both vectors and hosts vary in different regions. Colombia has a highly diverse mosquito fauna. Culex (Culex) bidens is reported here for the first time in Colombia. Because Cx. bidens Dyar and Knab and Cx. declarator Dyar and Knab share a close taxonomic history and because it is difficult to differentiate between them, a morphological and molecular comparison was performed. The male genitalia of three specimens of Cx. bidens from Colombia were mounted on microscope slides and morphologically compared with the male genitalia of Cx. declarator also from Colombia. In Cx. bidens, the individual teeth of the lateral plate are long, straight, laterally directed and sharply pointed; in Cx. declarator these teeth are robust, curved, with convex margins and bluntly rounded. Moreover, DNA was extracted from the same specimens and a fragment of the cytochrome c oxidase subunit I mitochondrial gene was amplified and sequenced. Neither Cx. bidens nor Cx. declarator were clustered in the Neighbour-joining topology, with K2P interspecific divergence between 0.15–1.45%. The circulation of Eastern Equine Encephalitis Virus in Colombia was reported since 1957 and Cx. bidens was suspected to be the vector of this virus during an epizootic in Argentina in 1988. Hybridization between species of the subgenus Culex has been demonstrated, hence the degree of reproductive isolation between Cx. bidens and Cx. declarator should be investigated, as well as their taxonomic status, because they only can be discriminated by a single male genitalic feature and not by nuclear or mitochondrial markers.

Estudios citogenéticos en híbridos triploides y tetraploides de Turnera krapovickasii, T. scabra y T. subulata (Passifloraceae, Turneroideae).

A fin de investigar las relaciones genómicas entre especies de la serie Turnera (x= 5), se lleva a cabo un programa de cruzamientos controlados y se obtuvieron numerosos híbridos. Entre las especies con flores amarillas de la serie Turnera, T. krapovickasii, T. scabra y T. subulata poseen citotipos diploide y autotetraploide. Hasta el momento se desconocen el grado de aislamiento reproductivo entre diploides y tetraploides y el grado de afinidad genómica de estas especies a nivel tetraploide. En este trabajo se presentan los resultados del análisis citogenético de los híbridos intraespecíficos triploides (2n=3x=15) de T. krapovickasii y T. scabra y los híbridos tetraploides (2n=4x=20) T. krapovickasii × T. scabra y T. subulata × T. scabra. En los híbridos triploides se hallaron 1-5 trivalentes y la viabilidad del polen fue 11,3% - 15,2%. En los híbridos tetraploides se encontraron células con hasta 5 cuadrivalentes y la viabilidad del polen fue 76% - 87,97%. Los resultados obtenidos demostraron que el aislamiento reproductivo entre diploides y tetraploides es incompleto y que los híbridos triploides no son completamente estériles. Asimismo, se evidenció que los genomios de T. krapovickasii, T. scabra y T. subulata serían más afines a nivel tetraploide que a nivel diploide.

Introgression between divergent corn borer species in a region of sympatry: Implications on the evolution and adaptation of pest arthropods.

The Asian corn borer, Ostrinia furnacalis, and European corn borer, O. nubilalis (Lepidoptera: Crambidae), cause damage to cultivated maize in spatially distinct geographies and have evolved divergent hydrocarbons as the basis of sexual communication. The Yili area of Xinjiang Uyghur Autonomous Region in China represents the only known region where O.furnacalis has invaded a native O. nubilalis range, and these two corn borer species have made secondary contact. Genetic differentiation was estimated between Ostrinia larvae collected from maize plants at 11 locations in Xinjiang and genotyped using high-throughput SNP and microsatellite markers. Maternal lineages were assessed by direct sequencing of mitochondrial cytochrome c oxidase subunit I and II haplotypes, and a high degree of genotypic diversity was demonstrated between lineages based on SNP genotypes. Furthermore, historical introgression was predicted among SNP genotypes only at sympatric locations in the Yili area, whereas in Xinjiang populations only O. furnacalis haplotypes were detected and no analogous introgressed genotypes were predicted. Our detection of putative hybrids and historical evidence of introgression defines Yili area as a hybrid zone between the species in normal ecological interactions and furthermore, might indicate that adaptive traits could spread even between seemingly divergent species through horizontal transmission. Results of this study indicate there may be a continuum in the degree of reproductive isolation between Ostrinia species and that the elegance of distinct and complete speciation based on modifications to the pheromone communication might need to be reconsidered.

Ophiostoma sugadairense, a new species in the Ophiostoma piceae complex associated with bark beetles infesting Japanese larch in Japan

A new species in the Ophiostoma piceae complex was isolated from bark beetles and beetle galleries infesting Japanese larch in Japan, and was described as Ophiostoma sugadairense. Phylogenetic analyses of the internal transcribed spacer of the nuclear ribosomal RNA genes region, partial β-tubulin, and elongation factor-1α genes revealed that O. sugadairense was clearly separated from other species of the O. piceae complex. The sexual morph of O. sugadairense morphologically resembled that of O. breviusculum, but the asexual morph was clearly distinguishable based on morphological characteristics of the synnemata, such as their color and shape. Mating compatibility tests indicated that there is some degree of reproductive isolation between O. sugadairense and O. piceae, O. breviusculum, and O. subalpinum.

Hybridization between two sister species of Bromeliaceae:Vriesea carinataandV. incurvata

Studies on interspecific gene flow and hybridization are important for understanding speciation processes and the movement of genes across species boundaries. In sympatric areas, hybridization between closely related species can be a common phenomenon and the degree of reproductive isolation (RI) among these species is closely related to the probability of hybrid formation. Here, using 14 nuclear microsatellites and two plastid DNA regions, we investigated the occurrence of natural hybridization in four sympatric populations of Vriesea carinata and V. incurvata. We also analysed four allopatric populations (two from each species), totalling 272 samples. These species share pollinators and have sequential flowering with a short period of overlap. Based on genetic assignment tests, 17 hybrids were detected in two of the four sympatric populations studied (8.29% of the individuals sampled). Our results showed low levels of nuclear (FST = 0.098) and high plastid genetic differentiation (FST = 0.601), with no haplotype sharing between parental species. The presence of plastid DNA haplotypes of both parental species in the hybrids indicated that hybridization probably occurred in both directions. We identified few hybrids, probably in consequence of some degree of RI between these species. Several RI barriers may be acting to maintain species integrity, temporal flowering differences being one important factor.

Biological aspects of crosses between Triatoma recurva (Stål), 1868 (Hemiptera: Reduviidae: Triatominae) and other members of the Phyllosoma complex.

The degree of reproductive isolation between Triatoma recurva (Stål) (Hemiptera: Reduviidae: Triatominae) and the six species of the genus Meccus plus T. mexicana (Herrich-Schaeffer) (Hemiptera: Reduviidae) was examined. Fertility and the segregation of morphological characteristics were examined in two generations of hybrids from crosses between these species. The percentage of couples with offspring (fertile) was low in the vast majority of sets of crosses, with the exception of that between T. recurva female and M. phyllosomus male. In all studied sets of crosses, no first- (F1) or second- (F2) generation individuals were morphologically similar to T. recurva but instead shared the morphology of the other parental species. A similar phenomenon was observed in the three successful sets of backcrosses. These results indicated that different recorded levels of reproductive fitness among T. recurva and the species of Meccus involved in this study, plus T. mexicana, are present and that they were apparently influenced by differing mechanisms of isolation. The presence of some degree of reproductive compatibility between studied triatomines of distinct genera (Meccus spp. and Triatoma spp.) reinforces the need for generic revision of the tribe Triatomini.

Mechanisms of reproductive isolation between incipient species in the Anopheles gambiae species complex

One of the greatest challenges in reducing malaria transmission in sub-Saharan Africa is the high degree of plasticity in the behaviour and ecology of members of the Anopheles gambiae complex, which includes the most efficient malaria vectors globally. For example, in areas of West Africa, sympatric populations of the M and S molecular forms of An. gambiae s.s. have diverged into recognized species (An. coluzzii Coetzee & Wilkerson sp.n. and the nominative species, An. gambiae s.s., respectively) through ecological speciation. Whereas the main breeding sites for An. gambiae s.s. continue to be temporary water bodies, An. coluzzii has become adapted to breeding well in irrigated fields, which has led to increases in relative abundance and duration of breeding seasons, and, hence, an enhancement of malaria transmission throughout the year. Thus, an increase in malaria prevalence is associated with a process of speciation that gave rise relatively rapidly to a new vector species highly adapted to a new breeding habitat created by changes in agricultural practice. To face this challenge, it is important that we understand the mechanisms by which this kind of speciation occurs. There is good evidence that speciation in this case has involved reproductive isolation, and, therefore, a five year study of sympatric populations of the M and S molecular forms in the Bobo Dioulasso area of Burkina Faso was undertaken to identify mechanisms that limit cross-matings. Mating in the An. gambiae complex generally occurs in swarms consisting mainly of males, which form at specific times of day at consistent sites defined by physical features in the environment, e.g., over wells, piles of wood, clearings in vegetation. More than 1,000 mating swarms were documented; a measurable degree of reproductive isolation could be explained by significant differences between species in seasonal abundance, characteristics of swarm sites and daily timing (circadian rhythm) of swarming. Although these factors each contribute to a reduced likelihood that males and females of ‘opposite’ molecular form encounter each other at close range in a mating swarm, both species were occasionally found in mixed swarms. All of the females caught in mixed swarms, however, had been inseminated by a conspecific male. Hence, further research is required to establish the close-range mechanism(s) of species recognition that seem to occur within swarms, such as auditory communication, that lead to assortative mating.