Linkage: from particulate to interactive genetics

Genetics was established on a strictparticulate conception of heredity. Geneticlinkage, the deviation from independentsegregation of Mendelian factors, was conceivedas a function of the material allocation of thefactors to the chromosomes, rather than to themultiple effects (pleiotropy) of discretefactors. Although linkage maps wereabstractions they provided strong support forthe chromosomal theory of inheritance. DirectCytogenetic evidence was scarce until X-rayinduced major chromosomal rearrangementsallowed direct correlation of genetic andcytological rearrangements. Only with thediscovery of the polytenic giant chromosomes inDrosophila larvae in the 1930s were thevirtual maps backed up by physical maps of thegenetic loci. Genetic linkage became a pivotalexperimental tool for the examination of theintegration of genetic functions in developmentand in evolution. Genetic mapping has remaineda hallmark of genetic analysis. The location ofgenes in DNA is a modern extension of thenotion of genetic linkage.     

The composition,diversity, and distribution of mosquito fauna (Diptera: Culicidae) in Kosovo

The aim of this study is to identify the mosquito species that currently exist and their distributions in Kosovo in order to determine current potential endemic zones and areas at a higher risk for future epidemics. These scientific data will be shared with public health authorities for implementing mosquito control programs. During a two‐year period of monitoring in 48 localities in 23 provinces in Kosovo, a total of 1,604 mosquitoes representing 13 species and six genera were collected and morphologically identified. Members of species complexes were also classified to species using DNA barcoding. In total, 13 species were identified with Culex pipiens s.l., the predominant species with an abundance rate of 39%. The remaining 12 species identified were grouped into five genera: Anopheles, Aedes, Coquillettidia, Culiseta, Uranotaenia, including species that are vectors of arboviruses in other parts of the world.     

A genetic linkage map for coho salmon (Oncorhynchus kisutch)

Construction of genetic linkage maps is an important first step for a variety of genomic applications, such as selective breeding in aquaculture, comparative studies of chromosomal evolution and identification of loci that have played key roles in the evolution of a species. Here we present a sex-specific linkage map for coho salmon. The map was constructed using 148 AFLP markers, 133 microsatellite loci and the phenotypic locus SEX . Twenty-four linkage groups spanning 287.4 cM were mapped in males, and 33 linkage groups spanning 429.7 cM were mapped in females. Several male linkage groups corresponded to two female linkage groups. The combination of linkage groups across both sexes appeared to characterize regions of 26 chromosomes. Two homeologous chromosomes were identified based on information from duplicated loci. Homologies between the coho and rainbow trout maps were examined. Eighty-six loci were found to form common linkage relationships between the two maps; these relationships provided evidence for whole-arm fissions, fusions and conservation of chromosomal regions in the evolution of these two species.     

Giemsa C-banding patterns in Aedes (Stegomyia) mosquitoes

Chromosomes from gonads of 14–24 h old pupae of nine species of Stegomyia mosquitoes have been examined using the Giemsa C-banding technique. The species studied were Aedes albopictus, A. polynesienis, A. scutellaris, A. alcasidi, A. seatoi, A. pseudalbopictus, A. melallicus, A. annandalei and A. vittatus. The diploid chromosome number of all species is six. All species possess C-bands in the centromeric regions of each of the three pairs of chromosomes. Besides, an intercalary C-band is present on the female determining (=m) chromosome but absent from the male—determining (= M) chromosome of all species except A. vittatus. In A. vittatus, the m and M chromosomes possess a terminal C-band. Thus, the nine species of Aedes analysed showed two distinct patterns of C-banding. —The evolution of heterochromatin patterns in various species is also discussed. The Giemsa C-banding technique should prove useful in studies of chromosomal speciation in culicine mosquitoes.     

Updated checklist of the mosquitoes (Diptera: Culicidae) of Belgium

Most information about the systematics and bioecology of Belgian mosquitoes dates back from before 1950, and only scattered information was produced during the last decades. In this paper we review and update the list of mosquito species recorded in Belgium, from first report (1908) to 2015. Six genera and 31 species were recorded so far, including 28 autochthonous species and three invasive alien species recently recorded in Belgium: Aedes albopictus (Skuse 1894), Ae. japonicus japonicus (Theobald 1901), and Ae. koreicus (Edwards 1917). The six genera are Anopheles (five species), Aedes (sixteen species), Coquillettidia (one species), Culex (four species), Culiseta (four species), and Orthopodomyia (one species).     

An updated checklist of the Culicidae (Diptera) of Morocco,with notes on species of historical and current medical importance

An updated checklist of the mosquito species (Diptera: Culicidae) recorded in Morocco from 1916 to 2016 is provided, including synonyms and synonymous usage for each species. Forty‐three species belonging to seven genera are recorded so far: Anopheles (9), Aedes (12) Coquillettidia (2), Culex (12), Culiseta (5), Orthopodomyia (1) and Uranotaenia (2). Traditional and equivalent names in the polyphyletic concept of Aedes are provided for the aedine species. The historical importance and current potential threat of mosquitoes to human health in Morocco is reviewed.     

Chromosome evolution in malaria mosquitoes

The genomic era offers excellent opportunities to improve our understanding of the genetic basis of mosquito adaptation, evolution, and competence to a pathogen. The availability of polytene chromosomes in anopheline mosquitoes makes them an excellent model system for studying genome organization, evolution, and function. Physical mapping facilitated the whole genome sequence assembly for the major malaria vector Anopheles gambiae and comparative genome mapping has determined types, patterns, and rates of chromosomal rearrangements in mosquito evolution. Together with sequencing projects, high-resolution physical mapping can shed light on mechanisms of chromosomal rearrangements and phylogenetic relation-ships among species.     

Analysis of genetic structure in soil populations of Rhizobium leguminosarum recovered from the USA and the UK

Although many studies have shown that animal-associated bacterial species exhibit linkage disequilibrium at chromosomal loci, recent studies indicate that both animal-associated and soil-borne bacterial species can display a nonclonal genetic structure in which alleles at chromosomal loci are in linkage equilibrium. To examine the situation in soil-borne species further, we compared genetic structure in two soil populations of Rhizobium leguminosarum bv. trifolii and two populations of R. leguminosarum bv. viciae from two sites in Oregon, with genetic structure in R. leguminosarum bv. viciae populations recovered from peas grown at a site in Washington, USA, and at a site in Norfolk, UK. A total of 234 chromosomal types (ET) were identified among 682 strains analysed for allelic variation at 13 enzyme-encoding chromosomal loci by multilocus enzyme electrophoresis (MLEE). Chi-square tests for heterogeneity of allele frequencies showed that the populations were not genetically uniform. A comparison of the genetic diversity within combined and individual populations confirmed that the Washington population was the primary cause of genetic differentiation between the populations. Each individual population exhibited linkage disequilibrium, with the magnitude of the disequilibrium being greatest in the Washington population and least in the UK population of R. leguminosarum bv. viciae. Linkage disequilibrium in the UK population was created between two clusters of 9 and 23 ETs, which, individually, were in linkage equilibrium. Strong linkage disequilibrium between the two major clusters of 8 and 12 ETs in the Washington population was caused by the low genetic diversity of the ETs within each cluster relative to the inter-cluster genetic distance. Because neither the magnitude of genetic diversity nor of linkage disequilibrium increased as hierarchical combinations of the six local populations were analysed, we conclude that the populations have not been isolated from each other for sufficient time, nor have they been exposed to enough selective pressure to develop unique multilocus genetic structure.     

Aedes albopictus oviposits with other Aedes species in artificial oviposition cups: a case study in Knox County,Tennessee, U.S.A.

Interspecific associations between two mosquito species can lead to effects such as competition, species displacement, and species stability. To better understand Aedes albopictus (Skuse) and other Aedes species, we monitored eggs in artificial oviposition cups (ovitraps) within Knox County, TN, U.S.A., during the 2016 and 2017 mosquito seasons. In 2016, one black and one white ovitrap were placed at 18 sites for 21 weeks, while in 2017 black and white ovitraps baited with grass‐infused or deionized water were placed at 11 sites for nine weeks. Eggs were identified to species and resulting counts were used to determine the degree of interspecific association using Cole's coefficients (C₇) and the degree of heterogeneity across space and time using generalized linear mixed models (GLMM). Here, Aedes mosquitoes are generally ovipositing in black cups with grass‐infused water, and Ae. albopictus eggs co‐occurred with other Aedes species more often than would be expected. Finding a positive significant interspecific association between Ae. albopictus and other Aedes eggs suggests that methods used to control Ae. albopictus may also control other Aedes mosquitoes. Finding that Ae. albopictus co‐occurs with other Aedes mosquitoes warrants additional research to evaluate outcomes associated with co‐occurrence within the study area.     

Genetic studies on hexokinase in the mosquitoAedes togoi

Hexokinases (EC 2.7.1.1) were genetically analyzed in the mosquitoAedes togoi by agar gel electrophoresis. Enzyme activity was observed anodally in one major banding region (HK-1) on the gel and in another faintly stained region (HK-2). A total of six bands was detected in the HK-1 region. All six bands could be detected in three body parts, head, thorax, and abdomen, of adults with different banding intensities. The third and fourth bands, numbered from the more anodal side, showed the broadest substrate specificity and the greatest enzyme activity throughout development. Genetic analysis of the six HK-1 bands was undertaken on the hypothesis of a single gene locus (or three extremely tightly linked loci). The analysis gave the following gene order:HK-1—4.2±1.8 (recombination units±SE)—To-2—Odh-2—29.5±2.5—sex (M/m)—s. A comparison is made of gene loci for hexokinases among the mosquito speciesCulex pipiens, Aedes aegypti, and this species, along with a comment on linkage relationships betweenHk andOdh (octanol dehydrogenase) loci in threeAedes species.This research was supported by National Institutes of Health Grant AI 16983 and a Grant-in-Aid for Co-operative Research (57340033) from the Ministry of Education, Science and Culture, Japan.     

Updated checklist of the mosquitoes (Diptera: Culicidae) of Croatia

Improvement of morphological and molecular identification methods allows the detection of new species of mosquitoes. The mosquito fauna of Croatia currently includes 52 species, belonging to eight genera, including Anopheles (12 species), Aedes (24 species), Coquillettidia (one species), Culex (seven species), Culiseta (six species), Orthopodomyia (one species), and Uranotaenia (one species). This is an updated checklist, which includes five new species found in Croatian mosquito fauna. Two of these are invasive mosquito species, Aedes albopictus (Skuse, 1895) and Aedes japonicus (Theobald 1901), which are spreading across Europe and Croatia. The other three species, Culex laticinctus (Edwards 1913), Culex torrentium (Martini 1925), and Anopheles daciae (Linton, Nicolescu & Harbach 2004) are autochthonous species which haven't been recorded so far. Since there are several more invasive species spreading across Europe, we assume that this is not the final list.     

Understanding the mechanism of Chikungunya virus vector competence in three species of mosquitoes

Chikungunya virus (CHIKV) is primarily transmitted by Aedes spp. mosquitoes. The present study investigated vector competence for CHIKV in Aedes aegypti and Aedes albopictus mosquitoes found in Madurai, South India. The role of receptor proteins on midguts contributing to permissiveness of CHIKV to Aedes spp. mosquitoes was also undertaken. Mosquitoes were orally infected with CHIKV DRDE‐06. Infection of midguts and dissemination to heads was confirmed by immunofluorescence assay at different time points. A plaque assay was performed from mosquito homogenates at different time points to study CHIKV replication. Presence of putative CHIKV receptor proteins on mosquito midgut epithelial cells was detected by virus overlay protein binding assay (VOPBA). The identity of these proteins was established using mass spectrometry. CHIKV infection of Ae. aegypti and Ae. albopictus midguts and dissemination to heads was observed to be similar. A plaque assay performed with infected mosquito homogenates revealed that CHIKV replication dynamics was similar in Aedes sp. mosquitoes until 28 days post infection. VOPBA performed with mosquito midgut membrane proteins revealed that prohibitin could serve as a putative CHIKV receptor on Aedes mosquito midguts, whereas an absence of CHIKV binding protein/s on Culex quinquefasciatus midguts can partially explain the non‐permissiveness of these mosquitoes to infection.     

Influence of competing and predatory invertebrate taxa on larval populations of mosquitoes in temporary ponds of wetland areas in Germany

Abundances of mosquito larvae and associated invertebrate communities were assessed in 27 temporary ponds during the spring season in wetland areas of Germany. Four genera of mosquitoes were identified: Aedes, Anopheles, Culex, and Culiseta. We focused our analyses on Aedes spp. because this genus was the most abundant (92% of total abundance) and frequently encountered mosquito (present in 65% of investigated sites). The abundance of Aedes spp. was negatively associated with the abundance of competitors for food, and to a lesser extent with those of intraguild predators and strict predators. The influence of these natural antagonists on larvae of Aedes was stronger in ponds with higher levels of dissolved oxygen (53 ± 4%) than in ponds with lower levels (16 ± 1%). The overall abundance of antagonists explained 42% of the variation in abundance of Aedes spp. at sites with higher levels of dissolved oxygen. Of this explained variation, competitors accounted for 34.7%, whereas the abundance of intraguild predators and strict predators accounted for only 6.8 and 0.5%, respectively. Therefore, the promotion of competing species might be an appropriate ecological approach for the control of Aedes spp. in temporary ponds in these areas.     

The actin loci in the genus Drosophila: establishment of chromosomal homologies among distantly related species by in situ hybridization

We have mapped by in situ hybridization the actin genes in selected, distantly related Drosophila species, using the 5C actin gene of D. melanogaster as a probe. In all species six dispersed actin loci were observed, probably corresponding to six genes, and they were similarly distributed among the chromosomes. In conjunction with previously available genetic and cytogenetic evidence, this consistent pattern of actin gene distribution reinforces the hypothesis that the chromosomal elements have maintained their essential identities throughout Drosophila evolution, and permits identification of these elements in very diverse species. Conservation of the actin loci also offers fixed points for the analysis of chromosomal inversions and other rearrangements.     

Individual inversions or their combinations: which is the main selective target in a natural population of Drosophila subobscura?

It is generally accepted that chromosomal inversions have been key elements in adaptation and speciation processes. In this context, Drosophila subobscura has been, and still is, an excellent model species due to its rich chromosomal polymorphism. In this species, many analyses from natural populations have demonstrated the adaptive potential of individual inversions (and their overlapped combinations, the so‐called arrangements). However, little information is available on the evolutionary role of combinations generated by inversions located in homologous and nonhomologous chromosomes. The aim of this research was to ascertain whether these combinations are also a target for natural selection. For this objective, we have studied the inversion composition of homologous and nonhomologous chromosomes from a D. subobscura sample collected in a well‐studied population, Mount Avala (Serbia). No significant deviation from H‐W expectations was detected, and when comparing particular karyotypic combinations, likelihood ratios close to 1 were obtained. Thus, it seems that for each pair of homologous chromosomes inversions no deviation from randomness was detected. Finally, no linkage disequilibrium was observed between inversions located in different chromosomes of the karyotype. For all these reasons, it can be assumed that, at the cytological level, the individual inversions rather than their combinations in different chromosomes are the main target of selection.     

A predicted microsatellite map of the passerine genome based on chicken-passerine sequence similarity

Abstract We present a predicted passerine genome map consisting of 196 microsatellite markers distributed across 25 chromosomes. The map was constructed by assigning chromosomal locations based on the sequence similarity between 550 publicly available passerine microsatellites and the draft chicken genome sequence published by the International Chicken Genome Sequencing Consortium. We compared this passerine microsatellite map with a recently published great reed warbler (Acrocephalus arundinaceus) linkage map derived from the segregation of marker alleles in a pedigree of a natural population. Twenty-four microsatellite markers were shared between the two maps, distributed across ten chromosomes. Synteny was maintained between the predicted passerine microsatellite map and the great reed warbler linkage map, confirming the validity and accuracy of our approach. Possible applications of the predicted passerine microsatellite map include genome mapping; quantitative trait locus (QTL) discovery; understanding heterozygosity-fitness correlations; investigating avian karyotype evolution; understanding microsatellite mutation processes; and for identifying loci conserved in multiple species, unlinked loci for use in genotyping sets and sex-linked markers.     

Habitat segregation among African treehole mosquitoes

Abstract. 1. The division of macrohabitats, seasons and microhabitats among mosquitoes which inhabit water-filled treeholes was investigated in a wet/dry seasonal environment near the coast of Kenya. Twenty-two species from four genera (Aedes fourteen, Culex three, Eretmapodites three, Toxorhynchites two) were identified from treeholes and bamboo traps. These included five which were more abundant in other microhabitats: leaf axils, fruit husks, or abandoned snail shells. 2. Trapping in different vegetation zones or heights in forests revealed that all seventeen species examined showed opposition preferences for one or more macrohabitats. Segregation of related species by macrohabitat was particularly apparent among members of the subgenus Stegomyia of Aedes which accounted for 57% of individuals identified from one locality. 3. Seasonal overlap was low between Aedes, which tracked rainfall patterns, and Culex and Eretmapodites. Aedes bromeliae differed from other Aedes found in treeholes in that maximal larval numbers did not follow heavy rainfall. 4. Negative associations between Aedes fulgens and two congeners which co-occur in the same forest and season were attributed to preferences for tree-holes of different sizes. Positive associations between Aedes aegypti and several other Aedes were reduced when root buttresses, a subclass of larger treeholes containing more species, were removed. 5. Separation of species by growth stages may be accomplished by two schemes of egg hatching which desynchronize larval cohorts in the same tree-hole: the early pattern of Stegomyia whereby 50–95% of all eggs hatch at first immersion, and the delayed pattern of other subgenera (Diceromyia, Finlaya, Pseudarmigeres) in which less than one-third of all eggs hatch at first immersion. 6. The number of wet treeholes changed seasonally. During one year five or six periods occurred when 50% or more of the censused holes contained sufficient water for larval survival. The number of individuals and species of mosquitoes in treeholes were positively correlated with treehole volume. In a comparison of treeholes and their mosquito fauna from two forests, treehole volume and mean number of species per hole differed significantly, but mosquito density did not. 7. Ecotonal trapping sites of mixed grasses, trees, and scrub yielded the highest mean numbers of species per sample, and traps placed outside a village, the lowest. Specific dominance was significantly less in the ecotone than elsewhere. The predatory larvae of Toxorhynchites brevipalpis in ecotonal and forested traps were not correlated with species richness.     

Diversified chromosomal characteristics in Centropyge fishes (Pomacanthidae, Perciformes)

We studied karyotypes and other chromosomal markers such as C-banded heterochromatin and Ag-stained nucleolus organizer regions (Ag-NORs), in seven Centropyge fishes (Pomacanthidae, Perciformes). These results revealed diversified chromosomal characteristics in Centropyge species. Three species had 2n = 48 chromosomes, whereas four species had 2n = 52 chromosomes. Fundamental numbers showed a large variation from 48 to 82, particularly in the species with 2n = 52 chromosomes. In all the species, Ag-NORs were located in a single chromosome pair and C-bands were mainly distributed in the centromeric regions of most chromosomes, as commonly seen in teleostean fishes. However, these chromosomal markers showed species-specific variations and provided us with useful information that could help us in understanding chromosomal evolution. On the basis of these chromosomal characteristics, we infer the process of chromosomal evolution, which according to us involves an increase in chromosome number from 2n = 48 to 2n = 52 through centric fission or other mechanisms, and in fundamental number through pericentric inversion. In particular, karyotypic evolution involving the increase in chromosome number is an unusual event in the evolution of higher teleostean groups. Handling editor: K. Martens     

Molecular approaches to mosquito parasite interactions

The success of vector borne disease transmission depends on the interplay between mosquito and pathogen. Understanding the genetic and molecular basis of refractoriness of mosquito may lead to novel disease control mechanisms. To complete the life cycle within the vector mosquito, a pathogen needs to overcome several physical barriers, such as the peritrophic matrix, midgut epithelium, or salivary glands. The immune response of the mosquito has to be neutralized or avoided. Genomic approaches are being employed to identify the genetic and molecular differences between selected strains of refractory and susceptible mosquitoes. Detailed molecular genetic maps based on restriction fragment length polymorphism (RFLP) and microsatellite (or simple sequence repeat) markers have been developed for two important vectors, Aedes aegypti and Anopheles gambiae, respectively. Recent success in genetic localization of quantitative trait loci controlling refractoriness/susceptibility of mosquitoes for Plasmodium and Brugian microfilariae provides a framework for further molecular characterization. Libraries of large genomic DNA inserts in bacterial or yeast artificial chromosomes (BACs and YACs) will facilitate physical mapping of the genetic loci controlling refractoriness. Identification of candidate refractory genes, and the cloning of other molecular markers for the mosquito immunity, provides tools for fruitful analysis of mosquito-parasite interactions. © 1997 Wiley-Liss, Inc.     

Experimental logging alters the abundance and community composition of ovipositing mosquitoes in the southern Appalachians

1. The loss of intact forest via logging can influence vector‐borne disease dynamics in part by altering the abundance or diversity of mosquito species. Using an experimental field approach, we characterised how two types of logging (clearcut and repeat‐entry shelterwood) affected temperate forest mosquito abundance and diversity in southwestern Virginia. 2.From May to September in 2008–2010, infusion‐baited gravid traps were used to collect ovipositing female mosquitoes across experimental forest plots that varied in logging treatment. Of the 29 680 collected adult female mosquitoes, the three dominant taxa captured were Aedes triseriatus (55%), Aedes japonicus (21%), and Culex pipiens/restuans (20%). 3. Logging treatment had a significant effect on the overall number of female mosquitoes caught per trap night, with lower average abundance of females on both logged treatments relative to two types of unlogged, control plots. When the three most abundant mosquito species were examined separately, logging treatment significantly influenced the abundance of both Aedes species, but did not significantly affect C. pipiens/restuans abundance. 4. Logging treatment did not influence the richness or diversity of mosquito species captured in gravid traps. However, logging treatment significantly altered the multivariate community composition of captured mosquitoes, an effect probably mediated by differential species‐specific impacts of logging on abundance. 5. Overall, the results of the present study suggest that the risk of arboviruses transmitted by container‐breeding Aedes species may be lower following a logging event in Appalachian forests because of reduced A. japonicus and A. triseriatus abundance with logging.