Single-locus Complementary Sex Determination Research Articles

Effects of Polyploidization on Locomotor and Flight Activity, and Body Size of Males in Japanese Bumblebee, Bombus ignitus

Polyploid animals are seen in the wild in only a few groups such as planaria, weevils, ants, wasps, and bees. The sex of bumblebees is determined by the single-locus complementary sex determination (sl-CSD) system resulting in females emerging from fertilized eggs and males (normal males, haploid) emerging from unfertilized eggs. However, diploid males (polyploids) emerged from fertilized eggs when the sex-determining locus become homozygous caused by inbreeding due to population contraction. To examine the effects of polyploidization on behavioral characteristics in bumblebee males, Bombus ignitus, haploid and diploid males were compared in locomotor activity, ease of staying inside the hive, and flight activity in a laboratory experiment. The results showed that diploid males had higher locomotor activity levels, were less likely to stay inside the hive, and had lower flight activity levels than haploid males. The reduced flight activity levels may lead to lower mating rates in the field. Furthermore, the body size of diploid males was smaller than that of haploid males. Diploid males of bumblebees have a low reproductive and produce diploid sperms to emerge sterile triploids, thus creating a considerable genetic load on the population. The behavioral and morphological characteristics of diploid males may unintentionally serve as a mechanism to prevent the emergence of sterile triploids.

Half-chromatid mutation as a possible cause of mosaic males and females in Hymenoptera and rare fertile male tortoiseshell cats.

Half-chromatid mutations occur when a single base change in a gamete is transmitted to the zygote, which, after DNA replication and cleavage, will result in a mosaic individual. These mutations will be passed on through the germ plasm and also may be expressed somatically. Half-chromatid mutation has been suggested to account for the observed lower frequency of males than expected for lethal X-linked recessive disorders in humans, such as Lesch-Nyhan syndrome, incontinentia pigmenti, and Duchene muscular dystrophy. Although attention has been paid to half-chromatid mutation in humans, it otherwise has been ignored. Here I show that half-chromatid mutation in haplodiploid organisms, such as Hymenoptera, has some interesting and important consequences: (i) since all genes follow the X-linked pattern of inheritance, half-chromatid mutations should be relatively easier to detect; (ii) recessive mutations of all viabilities may be expected; (iii) mosaics of both sexes are expected in haplodiploids with half-chromatid mutation; (iv) gynandromorphs could result from half-chromatid mutation at the sex-determination locus, in species with single-locus complementary sex-determination. Finally, half-chromatid mutation can account for the rare fertile male tortoiseshell phenotype observed in the domestic cat, Felis catus, and which still has not been fully accounted for by other mechanisms.

Ploidy determination in Bombus terrestris males: cost-efficiency comparison among different techniques

Diploid males represent a cost for the viability of populations in haplodiploid species with a single-locus complementary sex determination system. In such social species as bumble bees, their presence affects colony growth and reproduction, therefore, the detection of diploid males is an important aspect to implement conservation actions and for the breeding companies of this important pollinator. In this study, we applied and compared the effectiveness of morphological analyses (sperm count and testis area) with wing morphometrics (traditional and geometric) and genetic techniques (microsatellite analysis) to discriminate between diploid and haploid males originated from laboratory-reared B. terrestris colonies. Among the five techniques, microsatellite analyses and wing geometric morphometrics showed the highest efficiency in ploidy assignment and resulted in the most valuable methods according to a cost-effectiveness analysis.

Nest-departure behaviour of gynes and drones in the invasive yellowjacket Vespula germanica (Hymenoptera: Vespidae).

Inbreeding costs can be high in haplodiploid hymenopterans due to their particular mechanism of sex determination (i.e., single-locus complementary sex-determination system, sl-CSD), as it can lead to the production of sterile males. Therefore, mechanisms contributing to reduced inbred matings can be beneficial. In this sense, asynchronous nest departure of sibling drones and gynes could reduce kin encounters in social hymenopterans. Using six observation colonies, we determined under field conditions the nest departure behaviour of sibling reproductives of the social wasp Vespula germanica (Hymenoptera: Vespidae). We determined that sexuals leave the nests definitively and detected asynchronous departure not fixed to a particular caste at a seasonal scale in some colonies, as gynes or drones delayed their departure as a function of the departure of the opposite sex, depending on the colony. At a higher temporal resolution (i.e., within a day), we discovered that drones consistently began to leave nests 1 h before gynes and this difference was driven by those individuals that left on the same day as did the opposite-sex kin. Even though other mechanisms such as polyandry and differential dispersal could also be important at reducing inbred matings in the species, the observed departure patterns (i.e., in some colonies actually leave together with the opposite caste, while in others temporal segregation seems to occur) from nests could be complementary to the former and be important at reducing the negative effects of inbreeding in this invasive species.

Modeling diploid male dynamics in Hymenoptera: Effects of the number of alleles, dispersal by random walk and simple spatial structuring

Insects in the order Hymenoptera such as wasps, ants and bees exhibit a haplodiploid system of sexual determination in which females result from fertilized eggs and males from unfertilized eggs. Genetic information for sex determination is contained in a single multiallelic locus, so that the heterozygous individuals become females, and the hemizygous or homozygous individuals, haploid or diploid males, respectively. This mechanism is called single locus complementary sex determination (sl-CSD). While females and haploid males are regarded to be fertile, the opposite occurs for diploid males, so the significant production of diploid males (DM) may lead a population to an extinction scenario called the “diploid male vortex”. In the present work, we investigate the dynamics of populations of an organism with sl-CSD under various combinations of three parameters: male flight skills, number of sexual alleles and concentration of prohibited zones in a terrain represented by a square lattice. Sites of this lattice were randomly chosen as prohibited, where individuals could not pass, so the terrain was divided into islands of various sizes where individuals were confined. After a given simulation, the number of DM was measured. The main results that come from our simulations are: (i) the DM number depends more on the number of random steps in the male flight than on the number of alleles; (ii) the DM frequency in the small fragments becomes the same as the entire network when the limit of percolation is reached; (iii) the number of random steps explains the difference between the DM number found in the largest fragment and in the small fragments when the limit of percolation is reached.

Invasive population establishment of Asian honeybee Apis cerana

Bees, wasps, and ants are among the most highly invasive social insects. Theoretically, the single-locus complementary sex determination (sl-CSD) would hinder the establishment of invasive population with limited founders. Apis cerana has colonized many regions with anthropogenic movement, with devastating effects on the commercial A. mellifera. Here, we summarized the distribution and sex determination mechanism of A. cerana, the recent research results about the invasive A. cerana in Australia, and analyzed how they counteracted the handicap of their sex determination system by the queen's extreme polyandry, worker's reproduction, balancing selection and thelytoky. The knowledge reviewed here could be used as the reference to reveal the establishment mechanism of invasive populations of related social insects.

Complementary sex determination, inbreeding depression and inbreeding avoidance in a gregarious sawfly.

Although most Hymenoptera reproduce via arrhenotokous haplodiploidy, the underlying genetic mechanisms vary. Of these, the most widespread mechanism appears to be single-locus complementary sex determination (sl-CSD), in which individuals that are diploid and heterozygous at a sex-determining locus are female, and individuals that are homozygous or hemizygous are male. Because inbreeding increases the probability of producing diploid males, which are often sterile or inviable, sl-CSD can generate substantial inbreeding depression. To counteract this, Hymenoptera with traits that promote inbreeding, such as gregariousness, may evolve one or more of the following: inbreeding avoidance, functional diploid males or alternative sex determination mechanisms. Here, we investigate sex determination, inbreeding depression and inbreeding avoidance in Neodiprion lecontei, a gregarious, pine-feeding sawfly in the family Diprionidae. First, via inbreeding experiments and flow cytometry, we demonstrate that this species has CSD. By modeling expected sex ratios under different conditions, we also show that our data are consistent with sl-CSD. Second, via tracking survival in inbred and outbred families, we demonstrate that inbred families have reduced larval survival and that this mortality is partly attributable to the death of diploid males. Third, using a no-choice mating assay, we demonstrate that females are less willing to mate with siblings than nonsiblings. Together, these results suggest that inbreeding depression stemming from CSD has shaped mating behavior in N. lecontei. These results also set the stage for future comparative work that will investigate the interplay between sex determination, ecology and behavior in additional diprionid species that vary in larval gregariousness.

Genetic analyses and occurrence of diploid males in field and laboratory populations of Mastrus ridens (Hymenoptera: Ichneumonidae), a parasitoid of the codling moth

The parasitoid wasp Mastrus ridens (Hymenoptera: Ichneumonidae) is a particularly well-suited biological model to document the history and evolution of populations used in classical biological control, repeatedly moved from laboratory to laboratory worldwide and introduced in various environments. This specialist ectoparasitoid of the codling moth was first imported from Kazakhstan to the USA in the 1990’s, and then sent to Argentina, Chile and New Zealand. More recently, it was sent to Australia and France from other laboratory colonies, and imported again from field collections in Kazakhstan to Chile. Here, we used DNA sequencing to confirm the taxonomic identity of several populations used for biological control worldwide, and developed microsatellite markers for population genetics studies. A multiplex PCR amplifying 11 polymorphic markers was designed. These markers were used to compare the genetic diversity of laboratory and field populations and evaluate genetic differentiation between them. Results showed that laboratory populations with the longest rearing history had lower genetic diversity. Moreover, the genotyping of males with the markers revealed the occurrence of diploid males, which was further confirmed by flow cytometry, suggesting complementary sex determination (CSD) in this species. The percentage of diploid males in the populations ranged from 4% to 25% and were negatively correlated with diversity indices, which is consistent with a single-locus CSD and genetic bottlenecks in laboratory rearings. Molecular tools proved to be suitable and reliable for genetic diversity analyses in M. ridens, and should be implemented more frequently in classical biological control programs.

Diploid male dynamics under different numbers of sexual alleles and male dispersal abilities.

Insects in the order Hymenoptera (bees, wasps and ants) present an haplodiploid system of sexual determination in which fertilized eggs become females and unfertilized eggs males. Under single locus complementary sex-determination (sl-CSD) system, the sex of a specimen depends on the alleles at a single locus: when diploid, an individual will be a female if heterozygous and male if homozygous. Significant diploid male (DM) production may drive a population to an extinction scenario called "diploid male vortex". We aimed at studying the dynamics of populations of a sl-CSD organism under several combinations of two parameters: male flight abilities and number of sexual alleles. In these simulations, we evaluated the frequency of DM and a genetic diversity measure over 10,000 generations. The number of sexual alleles varied from 10 to 100 and, at each generation, a male offspring might fly to another random site within a varying radius R. Two main results emerge from our simulations: (i) the number of DM depends more on male flight radius than on the number of alleles; (ii) in large geographic regions, the effect of males flight radius on the allelic diversity turns out much less pronounced than in small regions. In other words, small regions where inbreeding normally appears recover genetic diversity due to large flight radii. These results may be particularly relevant when considering the population dynamics of species with increasingly limited dispersal ability (e.g., forest-dependent species of euglossine bees in fragmented landscapes).

Consequences of genetic incompatibility on fitness and mate choice: the male point of view

Hymenopterans under single-locus complementary sex determination (sl-CSD) face inbreeding costs due to this sex determination mode. Under sl-CSD, homozygote eggs at the sl-CSD locus usually develop into unviable or sterile diploid males. Production of such costly males increases when sib-mating happens because related individuals share half of their genome. In the hymenopteran Venturia canescens (a solitary parasitoid wasp), diploid males are sterile, leading to fitness costs through genetic incompatibility between parents. Whereas the costs of producing diploid males and behavioural strategies that would reduce such costs have been studied in females, the potential fitness costs faced by males have not. Here, we aimed to investigate fitness costs that males endure after a single sib-mating and tested whether they have the ability to avoid sib-mating through kin recognition. Our results show that males have a reduced fitness (i.e. they produce fewer daughters) when mating with their sibs. We also show that males have the ability to distinguish between non-sib and sib females (i.e. kin). They use chemical marks emitted by the females to discriminate kin from non-kin. We discuss the evolution of kin recognition in males in the context of mate choice for genetic compatibility. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2015, 114, 279–286.

Inbreeding depression in a parasitoid wasp with single-locus complementary sex determination.

Inbreeding and inbreeding depression are key processes in small or isolated populations and are therefore central concerns for the management of threatened or (re)introduced organisms. Haplodiploid species of the order Hymenoptera have a particular status with regard to inbreeding depression. Although recessive deleterious alleles that are expressed in males should be purged, an alternative form of inbreeding depression exists in species with single-locus complementary sex determination (sl-CSD). Under sl-CSD, genetically-related parents have a high probability of producing sterile sons instead of fertile daughters. In this article, we study inbreeding depression in Venturia canescens (Hymenoptera: Ichneumonidae), a parasitoid wasp with sl-CSD. We used a crossing design to manipulate relatedness according to three levels: within-family, between-family and between-population. For each level, several fitness components were measured on parents and female offspring. We found a 20% reduction in egg load at emergence for inbred crosses. Inbred crosses also yielded a higher proportion of males, as expected in a species with sl-CSD. Mating probability, presence of daughters among offspring, body size, symmetry and longevity were unaffected by inbreeding.

The genetic population structure of two socially parasitic ants: “the active slave-maker” Myrmoxenus ravouxi and the “degenerate slave-maker” M. kraussei

Slave-making ants exploit the societies of host ant species and are typically rare and patchily distributed. IUCN considers almost all slave-making ants as vulnerable, but solid data on their actual abundance are uncommon. Here we examine the genetic structure of populations of two species of the socially parasitic genus Myrmoxenus, which differ strongly in dispersal behavior and the occurrence of slave-raids. Microsatellite genotypes suggest strong differentiation even among neighboring populations of both species. FST-, G″ST-, and D-values were considerably higher in the “degenerate slave-maker” M. kraussei from Northern Italy than in the active slavemaker M. ravouxi from Southern Germany. This matches observations that sexuals of M. kraussei mate in their natal nest and queens disperse on foot while sexuals of M. ravouxi engage in mating and dispersal flights. Allelic richness was surprisingly high in both species and did not suggest recent bottlenecks, indicating that populations are larger and less vulnerable than expected from the difficulties of locating nests. Despite of considerable inbreeding, only very few diploid males were detected, supporting the view that at least in M. kraussei sex is not determined by single locus complementary sex determination unlike most other social Hymenoptera. The mismatch between the genetic and social structure of M. ravouxi colonies indicates occasionally fusion of slave-maker colonies in the field, mutual raids, or queen replacement. Complementary analyses of the host species of the two social parasites, Temnothorax unifasciatus and T. recedens, revealed low levels of population differentiation and confirmed the colony structure with a single, singly-mated queen.

Fertile diploid males in the ant Cataglyphis cursor: a potential cost of thelytoky?

Under the hymenopteran single-locus complementary sex-determination system, production of diploid males results from homozygosity at the sex-determiner locus. This arises when both parents transmit identical alleles at the locus to the offspring. In species reproducing asexually through thelytokous parthenogenesis, production of diploid males may also occur when the sex locus undergoes recombination and becomes homozygous in the offspring. Diploid males represent a substantial genetic load in hymenopteran populations because they often produce unviable sperm or sire sterile triploid female offspring. In the Mediterranean ant Cataglyphis cursor, the queen and workers can produce female offspring through automictic thelytokous parthenogenesis with central fusion, a mode of parthenogenesis that increases homozygosity. We report, for the first time, the presence of about 39 % of colonies producing adult diploid males (seven colonies out of 18). Overall, 8 % of adult males were diploid (12 diploid males out of the 146 males genotyped). Genotyping workers from the seven colonies producing diploid males showed that three diploid males were sons of queens and produced by thelytoky, six were probably sons of workers also produced by thelytoky and three were non-natal. Furthermore, the mating of a diploid male with two virgin queens in the laboratory led to the production of sterile triploid workers, which shows that diploid males in C. cursor are fertile, mate successfully and produce viable and functional but probably sterile female offspring. Because diploid males originate from thelytokous reproduction, they are only produced during sexual production and hence do not impair colony growth, which could explain why they are not removed at early brood stages.

Unexpectedly low frequencies of diploid males in an inbreeding parasitoid with complementary sex determination

In hymenopterans with single locus complementary sex determination, sex depends on the genotype at one polymorphic locus. Haploids are always male, while diploids are female when heterozygous and male when homozygous at the sex-determining locus. Brothers and sisters have a 50% chance of sharing a sex allele (i.e. of being ‘matched’), and hence half of all sibling matings are expected to produce diploid males at the expense of females. Nevertheless, observed frequencies of diploid males are often lower than predicted, as diploid males may succumb to pre-imaginal mortality, or because unmatched mates or sperm enjoy a competitive advantage. We counted diploid males in broods of the parasitoid wasp Cotesia glomerata sampled in the field, and in broods produced through controlled laboratory crosses. Consistently, the frequency of diploid males fell below expectations based upon the estimated occurrence of sibling mating. In the staged broods with diploid males, females made up a disproportionately large share of the diploids. Broods with and without diploid males were of similar size. Hence, the shortage of diploid males cannot be accounted for by differential pre-imaginal mortality alone. Instead, we postulate the existence of a mechanism that leads to preferential fertilization of eggs by sperm bearing unmatched alleles. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, ●●, ●●–●●.

Sex determination meltdown upon biological control introduction of the parasitoid Cotesia rubecula?

Natural enemies may go through genetic bottlenecks during the process of biological control introductions. Such bottlenecks are expected to be particularly detrimental in parasitoid Hymenoptera that exhibit complementary sex determination (CSD). CSD is associated with a severe form of inbreeding depression because homozygosity at one or multiple sex loci leads to the production of diploid males that are typically unviable or sterile. We observed that diploid males occur at a relatively high rate (8–13% of diploid adults) in a field population of Cotesia rubecula in Minnesota, USA, where this parasitoid was introduced for biological control of the cabbage white Pieris rapae. However, our laboratory crosses suggest two-locus CSD in a native Dutch population of C. rubecula and moderately high diploid males survival (approximately 70%), a scenario expected to produce low proportions of diploid males. We also show that courtship behavior of diploid males is similar to that of haploid males, but females mated to diploid males produce only very few daughters that are triploid. We use our laboratory data to estimate sex allele diversity in the field population of C. rubecula and discuss the possibility of a sex determination meltdown from two-locus CSD to effective single-locus CSD during or after introduction.

Tracing back the nascence of a new sex-determination pathway to the ancestor of bees and ants

In several Hymenoptera, sexual fate is determined by the allelic composition at the complementary sex-determiner locus, a sex-determination mechanism that can strongly affect population dynamics. To date, the molecular identification of complementary sex determiner has only been achieved in the honeybee, where the complementary sex-determiner gene was reported to have arisen from duplication of the feminizer gene. Strikingly, the complementary sex-determiner gene was also proposed to be unique to the honeybee lineage. Here we identify feminizer and complementary sex-determiner orthologues in bumble bees and ants. We further demonstrate that the duplication of feminizer that produced complementary sex determiner occurred before the divergence of Aculeata species (~120 Myr ago). Finally, we provide evidence that the two genes evolved concertedly through gene conversion, complementary sex-determiner evolution being additionally shaped by mosaic patterns of selection. Thus, the complementary sex-determiner gene likely represents the molecular basis for single locus-complementary sex determination in the Aculeata infra-order, and possibly, in the entire Hymenoptera order.

Csd alleles in the red dwarf honey bee (Apis florea, Hymenoptera: Apidae) show exceptionally high nucleotide diversity

Abstract The single locus complementary sex determination (sl-csd) gene is the primary gene determining the gender of honey bees (Apis spp.). While the csd gene has been well studied in the Western honey bee (Apis mellifera), and comparable data exist in both the Eastern honey bee (Apis cerana) and the giant honey bee (Apis dorsata), no studies have been conducted in the red dwarf honey bee, Apis florea. In this study we cloned the genomic region 3 of the A. florea csd gene from 60 workers, and identified 12 csd alleles. Analysis showed that similar to A. mellifera, region 3 of the csd gene contains a RS domain at the N terminal, a proline-rich domain at the C terminal, and a hypervariable region in the middle. However, the A. florea csd gene possessed a much higher level of nucleotide diversity, compared to A. mellifera, A. cerana and Apis dorsata. We also show that similar to the other three Apis species, in A. florea, nonsynonymous mutations in the csd gene are selectively favored in young alleles.

Females leave home for sex: natal dispersal in a parasitoid with complementary sex determination

The detrimental fitness effects of inbreeding are exacerbated in hymenopterans with single-locus complementary sex determination (sl-CSD) such as the parasitoid wasp Cotesia glomerata L. (Hymenoptera: Braconidae). Under sl-CSD, haploids are male, whereas diploids are female when heterozygous at the sex determination locus, but male when homozygous. Inbreeding leads to increased diploid male production. As diploid males can be of inferior fitness and are produced at the expense of females, species with sl-CSD are expected to avoid inbreeding. Natal dispersal of one or both sexes reduces the incidence of sibling matings. However, inbreeding avoidance is not necessarily the unique trigger of natal dispersal. Other factors, such as habitat variability and kin competition can also promote natal dispersal. Dispersal can be split into three stages, emigration, movement between patches and immigration, and revealing the ultimate causes of natal dispersal requires studying the proximate causes affecting these different stages. We monitored the postnatal behaviour of C. glomerata males and females in an experimental habitat that allowed us to track all three stages of dispersal and to manipulate the proximate factors triggering dispersal. None of the tested males, but more than one-third of the females dispersed when intersexual kin interactions were the only driving force of natal dispersal. Furthermore, we provide evidence that females dispersed to a new patch in search of mating opportunities. Our results demonstrate that inbreeding avoidance is the major cause of female natal dispersal in C. glomerata.

No evidence for increased extinction proneness with decreasing effective population size in a parasitoid with complementary sex determination and fertile diploid males.

BackgroundIn species with single locus complementary sex determination (sl-CSD), the sex of individuals depends on their genotype at one single locus with multiple alleles. Haploid individuals are always males. Diploid individuals are females when heterozygous, but males when homozygous at the sex-determining locus. Diploid males are typically unviable or effectively sterile, hence imposing a genetic load on populations. Diploid males are produced from matings of partners that share an allele at the sex-determining locus. The lower the allelic diversity at the sex-determining locus, the more diploid males are produced, ultimately impairing the growth of populations and jeopardizing their persistence. The gregarious endoparasitoid wasp Cotesia glomerata is one of only two known species with sl-CSD and fertile diploid males.ResultsBy manipulating the relatedness of the founders, we established replicated experimental populations of the parasitoid C. glomerata differing in their genetic effective size, and thus in allelic richness at the sex-determining locus and in the expected magnitude of diploid male production. Our long-term survey of population welfare and persistence did not provide evidence for increased proneness to population extinction with decreasing initial genetic effective population size. Most recorded surrogates of fitness nevertheless decayed over time and most experimental populations eventually went extinct, suggesting that the negative effects of inbreeding outweighed any premium from the fertility of diploid males.ConclusionsThe fertility of diploid males may have evolved as an adaptation prompted by the risk of extinction looming over small isolated populations of species with sl-CSD. However, fertility of diploid males does not negate the costs imposed by their production, and although it may temporarily stave off extinction, it is not sufficient to eradicate the negative effects of inbreeding.

CONSERVATION GENETICS OF NEOTROPICAL POLLINATORS REVISITED: MICROSATELLITE ANALYSIS SUGGESTS THAT DIPLOID MALES ARE RARE IN ORCHID BEES

Allozyme analyses have suggested that Neotropical orchid bee (Euglossini) pollinators are vulnerable because of putative high frequencies of diploid males, a result of loss of sex allele diversity in small hymenopteran populations with single locus complementary sex determination. Our analysis of 1010 males from 27 species of euglossine bees sampled across the Neotropics at 2–11 polymorphic microsatellite loci revealed only five diploid males at an overall frequency of 0.005 (95% CIs 0.002–0.010); errors through genetic nondetection of diploid males were likely small. In contrast to allozyme-based studies, we detected very weak or insignificant population genetic structure, even for a pair of populations >500 km apart, possibly accounting for low diploid male frequencies. Technical flaws in previous allozyme-based analyses have probably led to considerable overestimation of diploid male production in orchid bees. Other factors may have a more immediate impact on population persistence than the genetic load imposed by diploid males on these important Neotropical pollinators.