Reproductive Compatibility Research Articles

Extension of the distribution of Amblyomma triste Koch, 1844: Morphological and molecular confirmation of Morphotype I in Colombia

Ticks of the Amblyomma maculatum (Acari: Ixodidae) group are represented by three species: Amblyomma maculatum, Amblyomma tigrinum, and Amblyomma triste. This group is widely distributed throughout the Nearctic and Neotropical regions, from the United States of America (USA) to Argentina. They parasitize several vertebrates, including domestic mammals and humans. Nonetheless, the identification and distribution of the species within the group have been controversial but can be solved with integrative studies on their taxonomy and ecology. In Colombia, a Neotropical country from which up to 58 tick species have been reported, only Amblyomma maculatum Koch, 1844 (sensu stricto - s.s.) - morphotype II – has been molecularly confirmed. Here, specimens of the A. maculatum group from three departments of Colombia were examined by morphological and molecular analyses. The results confirmed the wide distribution of A. maculatum s.s. in Colombia. Furthermore, the presence of A. triste s.s. - morphotype I was confirmed for the first time at the molecular level, which broadens the distribution of this species in the American continent. Considering that both taxa are vectors of pathogens such as Rickettsia parkeri s.s., additional taxonomic, reproductive compatibility and niche distribution studies are needed, as well as the application of new molecular markers due to the controversial taxonomy. This will allow us to explore cryptic diversity within the recently established putative groups.

Morphology-Based Identification of Bemisia tabaci Cryptic Species Puparia via Embedded Group-Contrast Convolution Neural Network Analysis

The Bemisia tabaci species complex is a group of tropical–subtropical hemipterans, some species of which have achieved global distribution over the past 150 years. Several species are regarded currently as among the world’s most pernicious agricultural pests, causing a variety of damage types via direct feeding and plant-disease transmission. Long considered a single variable species, genetic, molecular and reproductive compatibility analyses have revealed that this “species” is actually a complex of between 24 and 48 morphologically cryptic species. However, determinations of which populations represent distinct species have been hampered by a failure to integrate genetic/molecular and morphological species–diagnoses. This, in turn, has limited the success of outbreak-control and eradication programs. Previous morphological investigations, based on traditional and geometric morphometric procedures, have had limited success in identifying genetic/molecular species from patterns of morphological variation in puparia. As an alternative, our investigation focused on exploring the use of a deep-learning convolution neural network (CNN) trained on puparial images and based on an embedded, group-contrast training protocol as a means of searching for consistent differences in puparial morphology. Fifteen molecular species were selected for analysis, all of which had been identified via DNA barcoding and confirmed using more extensive molecular characterizations and crossing experiments. Results demonstrate that all 15 species can be discriminated successfully based on differences in puparium morphology alone. This level of discrimination was achieved for laboratory populations reared on both hairy-leaved and glabrous-leaved host plants. Moreover, cross-tabulation tests confirmed the generality and stability of the CNN discriminant system trained on both ecophenotypic variants. The ability to identify B. tabaci species quickly and accurately from puparial images has the potential to address many long-standing problems in B. tabaci taxonomy and systematics as well as playing a vital role in ongoing pest-management efforts. [Aleyrodidae; entomology; Hemiptera; machine learning; morphometrics; pest control; systematics; taxonomy; whiteflies.]

Microbiome diversity and reproductive incompatibility induced by the prevalent endosymbiont Arsenophonus in two species of African cassava Bemisia tabaci whiteflies.

A minimum of 13 diverse whitefly species belonging to the Bemisia tabaci (B. tabaci) species complex are known to infest cassava crops in sub‐Saharan Africa (SSA), designated as SSA1‐13. Of these, the SSA1 and SSA2 are the predominant species colonizing cassava crops in East Africa. The SSA species of B. tabaci harbor diverse bacterial endosymbionts, many of which are known to manipulate insect reproduction. One such symbiont, Arsenophonus, is known to drive its spread by inducing reproductive incompatibility in its insect host and are abundant in SSA species of B. tabaci. However, whether Arsenophonus affects the reproduction of SSA species is unknown. In this study, we investigated both the reproductive compatibility between Arsenophonus infected and uninfected whiteflies by inter‐/intraspecific crossing experiments involving the sub‐group three haplotypes of the SSA1 (SSA1‐SG3), SSA2 species, and their microbial diversity. The number of eggs, nymphs, progenies produced, hatching rate, and survival rate were recorded for each cross. In intra‐specific crossing trials, both male and female progenies were produced and thus demonstrated no reproductive incompatibility. However, the total number of eggs laid, nymphs hatched, and the emerged females were low in the intra‐species crosses of SSA1‐SG3A+, indicating the negative effect of Arsenophonus on whitefly fitness. In contrast, the inter‐species crosses between the SSA1‐SG3 and SSA2 produced no female progeny and thus demonstrated reproductive incompatibility. The relative frequency of other bacteria colonizing the whiteflies was also investigated using Illumina sequencing of 16S rDNA and diversity indices were recorded. Overall, SSA1‐SG3 and SSA2 harbored high microbial diversity with more than 137 bacteria discovered. These results described for the first time the microbiome diversity and the reproductive behaviors of intra‐/inter‐species of Arsenophonus in whitefly reproduction, which is crucial for understanding the invasion abilities of cassava whiteflies.

Revisiting the hybridization processes in the Triatoma brasiliensis complex (Hemiptera, Triatominae): Interspecific genomic compatibility point to a possible recent diversification of the species grouped in this monophyletic complex.

Triatomines are hematophagous insects of great epidemiological importance, since they are vectors of the protozoan Trypanosoma cruzi, the etiological agent of Chagas disease. Triatoma brasiliensis complex is a monophyletic group formed by two subspecies and six species: T. b. brasiliensis, T. b. macromelasoma, T. bahiensis, T. juazeirensis, T. lenti, T. melanica, T. petrocchiae and T. sherlocki. The specific status of several species grouped in the T. brasiliensis complex was confirmed from experimental crossing and analysis of reproductive barriers. Thus, we perform interspecific experimental crosses between T. lenti and other species and subspecies of the T. brasiliensis complex and perform morphological analysis of the gonads and cytogenetic analysis in the homeologous chromosomes of the hybrids of first generation (F1). Besides that, we rescue all the literature data associated with the study of reproductive barriers in this monophyletic complex of species and subspecies. For all crosses performed between T. b. brasiliensis, T. b. macromelasoma, T. juazeirensis and T. melanica with T. lenti, interspecific copulas occurred (showing absence of mechanical isolation), hybrids were obtained, none of the male hybrids presented the phenomenon of gonadal dysgenesis and 100% pairing between the chromosomes homeologous of the hybrids was observed. Thus, we demonstrate that there are no pre-zygotic reproductive barriers installed between T. lenti and the species and subspecies of the T. brasiliensis complex. In addition, we demonstrate that the hybrids obtained between these crosses have high genomic compatibility and the absence of gonadal dysgenesis. These results point to reproductive compatibility between T. lenti and species and subspecies of the T. brasiliensis complex (confirming its inclusion in the complex) and lead us to suggest a possible recent diversification of the taxa of this monophyletic group.

Assisted gene flow using cryopreserved sperm in critically endangered coral

Assisted gene flow (AGF) is a conservation intervention to accelerate species adaptation to climate change by importing genetic diversity into at-risk populations. Corals exemplify both the need for AGF and its technical challenges; corals have declined in abundance, suffered pervasive reproductive failures, and struggled to adapt to climate change, yet mature corals cannot be easily moved for breeding, and coral gametes lose viability within hours. Here, we report the successful demonstration of AGF in corals using cryopreserved sperm that was frozen for 2 to 10 y. We fertilized Acropora palmata eggs from the western Caribbean (Curaçao) with cryopreserved sperm from genetically distinct populations in the eastern and central Caribbean (Florida and Puerto Rico, respectively). We then confirmed interpopulation parentage in the Curaçao-Florida offspring using 19,696 single-nucleotide polymorphism markers. Thus, we provide evidence of reproductive compatibility of a Caribbean coral across a recognized barrier to gene flow. The 6-mo survival of AGF offspring was 42%, the highest ever achieved in this species, yielding the largest wildlife population ever raised from cryopreserved material. By breeding a critically endangered coral across its range without moving adults, we show that AGF using cryopreservation is a viable conservation tool to increase genetic diversity in threatened marine populations.

Hybridization and species boundaries between three sympatric bromeliads from the Brazilian Atlantic Forest

Abstract Recently diversified taxa commonly do not show complete reproductive isolation, and thus they represent an interesting model for studying the occurrence of hybridization and its influence on the integrity of species. In this study, we aim to elucidate the occurrence of hybridization and discuss species boundaries between three Aechmea spp. (Bromeliaceae) from southern Brazil. To achieve these aims, we used ten nuclear and six plastid microsatellite loci. We also investigated morphological and ecological differentiation and reproductive compatibility through interspecific cross-pollination experiments with these species. Molecular analyses confirmed the occurrence of hybridization between the three species, but interspecific gene flow was higher between A. comata and A. kertesziae than between A. comata and A. caudata. These Aechmea spp. showed morphological differentiation, and the results of the artificial interspecific crosses indicated that they can intercross. Aechmea caudata and A. kertesziae show distinct microhabitat preferences, whereas A. comata thrives in a microhabitat that is also inhabited by the other species. Despite the potential reproductive barriers being weak, especially those between A. comata and A. kertesziae, they may maintain the boundaries between these species and their integrity in sympatric populations.

Trends in taxonomy of Triatomini (Hemiptera, Reduviidae, Triatominae): reproductive compatibility reinforces the synonymization of Meccus St\xe5l, 1859 with Triatoma Laporte, 1832

BackgroundMeccus' taxonomy has been quite complex since the first species of this genus was described by Burmeister in 1835 as Conorhinus phyllosoma. In 1859 the species was transferred to the genus Meccus and in 1930 to Triatoma. However, in the twentieth century, the Meccus genus was revalidated (alteration corroborated by molecular studies) and, in the twenty-first century, through a comprehensive study including more sophisticated phylogenetic reconstruction methods, Meccus was again synonymous with Triatoma. Events of natural hybridization with production of fertile offspring have already been reported among sympatric species of the T. phyllosoma subcomplex, and experimental crosses demonstrated reproductive viability among practically all species of the T. phyllosoma subcomplex that were considered as belonging to the genus Meccus, as well as between these species and species of Triatoma. Based on the above, we carried out experimental crosses between T. longipennis (considered M. longipennis in some literature) and T. mopan (always considered as belonging to Triatoma) to evaluate the reproductive compatibility between species of the T. phyllosoma complex. In addition, we have grouped our results with information from the literature regarding crosses between species that were grouped in the genus Meccus with Triatoma, in order to discuss the importance of experimental crosses to confirm the generic reorganization of species.ResultsThe crosses between T. mopan female and T. longipennis male resulted in viable offspring. The hatching of hybrids, even if only in one direction and/or at low frequency, demonstrates reproductive compatibility and homeology between the genomes of the parents.ConclusionConsidering that intergeneric crosses usually do not result in viable offspring in Triatominae, the reproductive compatibility observed between the T. phyllosoma subcomplex species considered in the Meccus genus with species of the Triatoma genus shows that there is “intergeneric” genomic compatibility, which corroborates the generic reorganization of Meccus in Triatoma.Graphical

Prevalence and intensity of avian malaria in a quail hybrid zone.

Hybrid zones have been described as natural laboratories by researchers who study speciation and the various mechanisms that may affect gene flow. The evolutionary consequences of hybridization depend not only on reproductive compatibility between sympatric species, but also on factors like vulnerability to each other's predators and parasites. We examined infection patterns of the blood parasite Haemoproteus lophortyx, a causative agent of avian malaria, at a site in the contact zone between California quail (Callipepla californica) and Gambel's quail (C. gambelii). Controlling for the potential influence of sex and year, we tested whether species identity predicted infection status and intensity. We found that infection prevalence was lower in California and hybrid quail compared with Gambel's quail. However, infected California and hybrid quail had higher infection intensities than Gambel's quail. California and hybrid quail exhibited no significant differences in prevalence or intensity of infection. These findings suggest that infection by H. lophortyx has the potential to influence species barrier dynamics in this system; however, more work is necessary to determine the exact evolutionary consequences of this blood parasite on hybridization.

The genus Phagocata Leidy (Platyhelminthes, Tricladida) in Israel, a new species of Phagocata from Lake Kinneret, and an emended description of Dugesia salina

The genus Phagocata (Platyhelminthes, Tricladida) is represented in Israel by two pigmented species, namely, Phagocata armeniaca (Komárek, 1916), reported previously from Armenia, the Caucasus and eastern Turkey, and more recently known from the constantly cold headwaters of the River Jordan in northern Israel, and Phagocata punctata sp. nov., which was found inhabiting the littoral of the south-eastern shore of Lake Kinneret where there are large seasonal temperature fluctuations. Several samples of both species were collected in different seasons and raised in the laboratory at various temperatures and their habitats, morphology, karyology (2n = 34) and breeding behaviour were compared. The results of cross-breeding experiments are given, with most of the F1 offspring showing a range of patterns of colouration, gut branching and fecundity which were intermediate to those of the parent species. However, crossbreeding also resulted in a small proportion of abnormalities among the F1, and especially the F2, generations, indicating an incomplete reproductive compatibility, but supporting the hypothesis of their common origin. An emended description of Dugesia salina (Whitehouse, 1914), is also given together with karyological data (2n = 16) from material collected from a saline spring, En Sheva (Tabgha), situated on the north-western shore of Lake Kinneret.

Dimorphism, Polyploidy, and Genetic Diversity in the Australian Endemic Lycium australe (Solanaceae)

Premise of research. Whole-genome duplication is often associated with the evolution of dimorphic sexual systems; however, the association is not universal, and the evolutionary pathway(s) underlying the association remain unresolved. The genus Lycium has been demonstrated as a useful system for investigating ploidy and sexual system variation. Here, we describe ploidy and floral variation in L. australe and document genetic diversity and dimorphism across its range.Methodology. Sexual system and size dimorphism were assessed using morphological measurements of 486 flowers. Flow cytometry was used to determine DNA content and ploidy level for 167 plants from across the species range. Using the plastid region trnH-psbA and restriction site–associated DNA sequencing (RADseq), we explored the genetic structure of the species, including an examination of associations of ploidy and sexual system with genetic structure.Pivotal results. We find correlated variation in ploidy level, sexual system, and genetic structure in L. australe. Both gynodioecy and dioecy are documented and are associated with diploidy and polyploidy, respectively. Floral size dimorphism is also found among morphs in both sexual systems. We find significant genetic structure associated with ploidy level and geography based on genome-wide single-nucleotide polymorphisms and plastid haplotypes.Conclusions. We show that L. australe has associated polymorphism in sexual system and ploidy, but the association is an exception to the pattern in the genus. Genetic and ploidy variation in the species suggests potential reproductive barriers and ecological differentiation between cytotypes of differing ploidies. The widespread parapatric distributions of diploid and polyploid L. australe make it an ideal system for studying questions related to reproductive compatibility, ecological differentiation, and polyploid speciation.

Reproductive compatibility in Capsicum is not necessarily reflected in genetic or phenotypic similarity between species complexes.

Wild relatives of domesticated Capsicum represent substantial genetic diversity and thus sources of traits of potential interest. Furthermore, the hybridization compatibility between members of Capsicum species complexes remains unresolved. Improving our understanding of the relationship between Capsicum species relatedness and their ability to form hybrids is a highly pertinent issue. Through the development of novel interspecific hybrids in this study, we demonstrate interspecies compatibility is not necessarily reflected in relatedness according to established Capsicum genepool complexes. Based on a phylogeny constructed by genotyping using simple sequence repeat (SSR) markers and with a portion of the waxy locus, and through principal component analysis (PCA) of phenotypic data, we clarify the relationships among wild and domesticated Capsicum species. Together, the phylogeny and hybridization studies provide evidence for the misidentification of a number of species from the World Vegetable Center genebank included in this study. The World Vegetable Center holds the largest collection of Capsicum genetic material globally, therefore this may reflect a wider issue in the misidentification of Capsicum wild relatives. The findings presented here provide insight into an apparent disconnect between compatibility and relatedness in the Capsicum genus, which will be valuable in identifying candidates for future breeding programs.

Nectar robbing and plant reproduction: an interplay of positive and negative effects

Nectar robbers are animals that extract nectar through holes made in floral tissues. This behaviour has a wide spectrum of consequences for the plant that range from negative, to neutral, to positive according to life history traits of the interacting organisms and the ecological mechanisms involved. Lonicera etrusca has long tubular flowers producing large quantities of nectar and undergoing high levels of nectar robbing. Despite this, robbing seems to have neutral effects on the reproduction of the species. The aim of this work is to understand why this pollinator‐dependent plant is not affected by an interaction that is commonly detrimental for plants. To achieve this goal we try to answer two questions: 1) do nectar robbers deposit suitable pollen for plant reproduction after a visit? And, 2) is the visitation rate of legitimate visitors affected by nectar robbing? We experimentally studied the details of the reproductive compatibility system of the plant and compared pollen tube growth after single visits of the main nectar robbers. We made observations on plants of three populations to study the effect of robbers on the visitation rates of legitimate foragers. The results revealed that nectar robbers effectively perform cross‐pollination. Visitation rates of legitimate visitors are reduced by nectar robbing. The net neutral effect observed for L. etrusca is the result of two processes: on one side the direct positive effect of nectar robbers acting as pollinators, and on the other, a negative indirect effect caused by the decrease in visitation rates of legitimate visitors. Since the number of ovules in this species is low, both legitimate visitors and robbers contribute to an efficient cross‐pollination. This example illustrates how neutral effects for the plant can arise as the final result of a complex interaction between plants, legitimate visitors and robbers.

DISTATIS: A Promising Framework to Integrate Distance Matrices in Molecular Phylogenetics.

Molecular phylogenetic algorithms frequently disagree with the approaches considering reproductive compatibility and morphological criteria for species delimitation. The question stems if the resulting species boundaries from molecular, reproductive and/or morphological data are definitively not reconcilable; or if the existing phylogenetic methods are not sensitive enough to agree morphological and genetic variation in species delimitation. We propose DISTATIS as an integrative framework to combine alignment-based (AB) and alignment-free (AF) distance matrices from ITS2 sequences/structures to shed light whether Gelasinospora and Neurospora are sister but independent genera. We aimed at addressing this standing issue by harmonizing genus-specific classification based on their ascospore morphology and ITS2 molecular data. To validate our proposal, three phylogenetic approaches: i) traditional alignment-based, ii) alignment-free and iii) novel distance integrative (DI)-based were comparatively evaluated on a set of Gelasinospora and Neurospora species. All considered species have been extensively characterized at both the morphological and reproductive levels and there are known incongruences between their ascospore morphology and molecular data that hampers genus-specific delimitation. Traditional AB phylogenetic analyses fail at resolving the Gelasinospora and Neurospora genera into independent monophyletic clades following ascospore morphology criteria. In contrast, AF and DI approaches produced phylogenetic trees that could properly delimit the expected monophyletic clades. The DI approach outperformed the AF one in the sense that it could also divide the Neurospora species according to their reproduction mode.

Fitness and mating compatibility of Chrysodeixis includens (W.) (Lepidoptera: Noctuidae) populations collected in different provinces and crops in Argentina.

Chrysodeixis includens, is an important Plusiinae pest that feeds on soybean, beans, and cotton. This species has a genetic structure determined by molecular markers in populations collected in soybean crops in Brazil and has developed resistance to insecticides. This had led to an increased interest in the genetic diversity of its populations in Argentina. This study aimed to characterize the fitness parameters and mating compatibility of geographically distant C. includens populations and on different host plants in Argentina. Intra- and inter-population crosses revealed that C. includens populations from Tucumán and Santiago del Estero provinces did not present any evidence of prezygotic and postzygotic incompatibilities, suggesting that both populations collected from soybean plants belonged to a single wide-ranging species. Prezygotic and postzygotic incompatibilities were detected between populations collected from different crops (soybean and bean). These incompatibilities may be caused by the different nutrients that each plant species renders, or the specific pest control strategies which may affect C. includens behavior. Studies of the genetic structure of C. includens populations based on mating isolation on other crops, as well as of the impact of management strategies on the specie, will increase understanding of the reproductive compatibility revealed by the present study.

Reproductive compatibility of Sorbocotoneaster with mountain ash species and varieties that are promising for breeding in Siberia

In Russia, in the Republic of Yakutia, there is the only point in the world where natural intergeneric hybridization of species of the genera Sorbus and Cotoneaster is observed, resulting in the formation of a new hybridogenic genus ×Sorbocotoneaster with a rich polymorphism of parental genera traits. For the selection of mountain ash in Siberia, it is promising to involve Sorbocotoneaster genotypes in artificial hybridization as sources of a complex of economically valuable traits, including winter hardiness, short stature, and self-fertility. Artificial hybridization of the tetraploid selective form Sorbocotoneaster with selected forms and varieties of Sorbus sibirica, S. aucuparia, S. sambucifolia and their hybrids allows obtaining viable hybrid offspring in combinations where Sorbocotoneaster is used as a pollen donor.

Molecular systematics and phylogeography of a widespread Neotropical avian lineage: evidence for cryptic speciation with protracted gene flow throughout the Late Quaternary

AbstractHere we use an integrative approach, including coalescent-based methods, isolation–migration and species distribution models, to infer population structure, divergence times and diversification in the two species of the genus Cymbilaimus (Aves, Thamnophilidae). Our results support a recent and rapid diversification with both incomplete lineage sorting and gene flow shaping the evolutionary history of Cymbilaimus. The spatio-temporal pattern of cladogenesis suggests that Cymbilaimus originated in the north/western portion of cis-Andean South America and then diversified into the Brazilian Shield and Central America after consolidation of the modern Amazonian drainage and the Andean range. This evolutionary scenario is explained by cycles of range expansion and dispersal, followed by isolation, and recurrent gene flow, during the last 1.2 Myr. Our results agree with those recently reported for other closely related suboscine lineages, whereby the window of introgression between closely related taxa remains open for up to a few million years after their original split. In Cymbilaimus, introgression was recurrent between C. lineatus and C. sanctaemariae, even after they acquired vocal and ecological differentiation, supporting the claim that at least in Neotropical suboscines, full reproductive compatibility may take millions of years to evolve and cannot be interpreted as synonymous with a lack of speciation.

Honeybush (Cyclopia spp.) anther-stigma distance and intraspecies cross compatibilities

Species of Cyclopia are used to produce honeybush tea, a uniquely South African beverage that is rapidly growing in popularity locally and abroad. In response to warnings about ecological sustainability, the honeybush industry is transitioning from a wild-harvested source model to one relying primarily on commercial farming. To this end, honeybush genotypes are needed with improved homogeneity, sensory qualities, productivity, vigour and resistance to such pest and disease conditions as may arise in massed plantings. In support of existing honeybush breeding programmes, studies have been undertaken to answer pressing questions about honeybush reproductive anatomy and sexual compatibility. Anther-stigma distance was measured within and between honeybush genotypes in order to rule out flower heteromorphism as a cause for low seed set following hand pollination. Continuous variation in anther-stigma distance was observed for the homomorphic, leguminous flowers, suggesting the existence of a mixed-mating system. Diallel crosses were then performed by hand pollination between genotypes within three honeybush species. All three species were found to be partially self-compatible, with large variations in self-incompatibility between genotypes within species. This study provides practical recommendations with regard to the sexual propagation of select honeybush species and expands the platform of scientific research available to honeybush breeders.

Reproductive incompatibility between Amblyomma maculatum (Acari: Ixodidae) group ticks from two disjunct geographical regions within the USA.

The Amblyomma maculatum Koch group of ixodid ticks consists of three species: A. maculatum, A. triste, and A. tigrinum. However, since Koch described this group in 1844, the systematics of its members has been the subject of ongoing debate. This is especially true of A. maculatum and A. triste; recent molecular analyses reveal insufficient genetic divergence to separate these as distinct species. Further confounding this issue is the discovery in 2014 of A. maculatum group ticks in southern Arizona (AZ), USA, that share morphological characteristics with both A. triste and A. maculatum. To biologically evaluate the identity of A. maculatum group ticks from southern Arizona, we analyzed the reproductive compatibility between specimens of A. maculatum group ticks collected from Georgia (GA), USA, and southern Arizona. Female ticks from both Arizona and Georgia were mated with males from both the Georgia and Arizona Amblyomma populations, creating two homologous and two heterologous F1 cohorts of ticks: GA ♀/GA ♂, AZ ♀/AZ ♂, GA ♀/AZ ♂, and AZ ♀/GA ♂. Each cohort was maintained separately into the F2 generation with F1 females mating only with F1 males from their same cohort. Survival and fecundity parameters were measured for all developmental stages. The observed survival parameters for heterologous cohorts were comparable to those of the homologous cohorts through the F1 generation. However, the F1 heterologous females produced F2 egg clutches that did not hatch, thus indicating that the Arizona and Georgia populations of A. maculatum group ticks tested here represent different biological species.

Reproductive compatibility among populations and host-associated lineages of the common bed bug (Cimex lectularius L.).

As populations differentiate across geographic or host‐association barriers, interpopulation fertility is often a measure of the extent of incipient speciation. The bed bug, Cimex lectularius L., was recently found to form two host‐associated lineages within Europe: one found with humans (human‐associated, HA) and the other found with bats (bat‐associated, BA). No unequivocal evidence of contemporary gene flow between these lineages has been found; however, it is unclear whether this is due to an inability to produce viable “hybrid” offspring. To address this question and determine the extent of compatibility between host‐associated lineages, we set up mating crosses among populations of bed bugs based on both their host association (human—HA vs. bat—BA) and geographic origin (North America vs. Europe). Within‐population fecundity was significantly higher for all HA populations (>1.7 eggs/day) than for BA populations (<1 egg/day). However, all within‐population crosses, regardless of host association, had >92% egg hatch rates. Contrary to previous reports, in all interlineage crosses, successful matings occurred, fertile eggs were oviposited, and the F1 “hybrid” generation was found to be reproductively viable. In addition, we evaluated interpopulation genetic variation in Wolbachia among host‐associated lineages. We did not find any clear patterns related to host association, nor did we observe a homogenization of Wolbachia lineages across populations that might explain a breakdown of reproductive incompatibility. These results indicate that while the HA and BA populations of C. lectularius represent genetically differentiated host‐associated lineages, possibly undergoing sympatric speciation, this is in its incipient stage as they remain reproductively compatible. Other behavioral, physiological, and/or ecological factors likely maintain host‐associated differentiation.

The puzzling mitochondrial phylogeography of the black soldier fly (Hermetia illucens), the commercially most important insect protein species

BackgroundThe black soldier fly (Diptera: Stratiomyidae, Hermetia illucens) is renowned for its bioconversion ability of organic matter, and is the worldwide most widely used source of insect protein. Despite varying extensively in morphology, it is widely assumed that all black soldier flies belong to the same species, Hermetia illucens. We here screened about 600 field-collected and cultured flies from 39 countries and six biogeographic regions to test this assumption based on data for three genes (mitochondrial COI, nuclear ITS2 & 28S rDNA) and in order to gain insights into the phylogeography of the species.ResultsOur study reveals a surprisingly high level of intraspecific genetic diversity for the mitochondrial barcoding gene COI (divergences up to 4.9%). This level of variability is often associated with the presence of multiple species, but tested nuclear markers (ITS2 and 28S rDNA) were invariant and fly strain hybridization experiments under laboratory conditions revealed reproductive compatibility. COI haplotype diversity is not only very high in all biogeographic regions (56 distinct haplotypes in total), but also in breeding facilities and research centers from six continents (10 haplotypes: divergences up to 4.3%). The high genetic diversity in fly-breeding facilities is mostly likely due to many independent acquisitions of cultures via sharing and/or establishing new colonies from field-collected flies. However, explaining some of the observed diversity in several biogeographic regions is difficult given that the origin of the species is considered to be New World (32 distinct haplotypes) and one would expect severely reduced genetic diversity in the putatively non-native populations in the remaining biogeographic regions. However, distinct, private haplotypes are known from the Australasian (N = 1), Oriental (N = 4), and the Eastern Palearctic (N = 4) populations. We reviewed museum specimen records and conclude that the evidence for introductions is strong for the Western Palearctic and Afrotropical regions which lack distinct, private haplotypes.ConclusionsBased on the results of this paper, we urge the black soldier fly community to apply molecular characterization (genotyping) of the fly strains used in artificial fly-breeding and share these data in research publications as well as when sharing cultures. In addition, fast-evolving nuclear markers should be used to reconstruct the recent invasion history of the species.