Chromosomal evidence of hemiclonal and all-paternal offspring production in Bacillus rossius-grandii benazzii (Insecta Phasmatodea)

The stick insects Bacillus rossius-grandii benazzii and B. rossius-grandii grandii naturally reproduce by hybridogenesis and androgenesis. The hybrid karyotype of the former (2n=35, XX female; 34, X0 male) clearly sums up a B. rossius haploset (r) with n=18 and a B. grandii benazzii one (gb) with n=17. The two sets keep the parental features for C-heterochromatin amount (much larger in the gb complement) and satellites/NORs (nuclear organizer regions) (more numerous and variably located in the r set); hybridogenetically produced males always show severely impaired gametogenesis and are therefore sterile, whereas hybridogenetic females are fertile. Reproductive, karyological and cytogenetical properties of the hybridogenetic system have been exploited to obtain the chromosomal evidence of whole haploset exchanges. In progenies obtained by crossing B. rossiusgrandii benazzii females to B. rossius males with either standard or repatterned (with Robertsonian fusions) karyotypes, there has always been complete agreement between electrophoretically genotyped and karyologically analyzed hybridogenetic offspring: the unassorted maternal r haploset (r ^m) is transmitted and the gb ^m haploset replaced by that of the fathering male (r ^p), thus evidencing the hemiclonal reproduction and the new r ^m-r ^p chromosomal constitution. New karyotype traits of the offspring relate to chromosome number (2n=36, female; 35, male), C-heterochromatin pattern (the heterochromatin-rich gb haploset completely disappears) and satellite/NOR features (corresponding to r ^m plus r ^p locations). The same crosses also produce genetically and chromosomally all-paternal descendants (androgenetics), of both sexes and fully fertile, with an r ^p r ^p structure. These androgenetic progeny show segregation for alleles and chromosomes at which fathering males are heterozygous: it was therefore possible to demonstrate that androgenetics can derive from syngamy of two sperm nuclei, of the several present in the polyspermic hybridogenetic egg. The production of androgenetics from field fertilized females of B. rossius-grandii benazzii, B. rossius-grandii grandii and parthenogenetic Bacillus whitei (=B. rossius/grandii grandii) suggests the occurrence of unsuspected relationships between hybrids and their parental species, so that the hybrids cannot be simply considered as sexual parasites. Furthermore, there is a suggestion of evolution of parthenogenetic clonal species from selection of initially hybridogenetic strains. The ability to produce uniparental progeny naturally from the spermatic genome may open a new field of investigation on genomic imprinting.     

Allozyme analysis and phyletic relationships of two new stick-insects from north-west Sicily: Bacillus grandii benazzii and B. rossius-grandii benazzii (Insecta Phasmatodea)

Two new Sicilian stick-insects have been discovered within the genus Bacillus. Evidence concerning the subspecific differentiation of Bacillus grandii benazzii and the hybrid constitution of B. rossius-grandii benazzii by means of allozyme analysis is given, and is consistent with morphological and karyological data on the differentiation of these two north-west Sicilian morphs from south-eastern B. g. grandii and B. whitei. B. g. benazzii is more polymorphic than B. g. grandii and B. rossius-g. benazzii embodies its genetic variability, thus differing sharply from the south-eastern parallel hybrid B. whitei ( = B. rossius × B. g. grandii). Reproductive mechanisms also appear to be different in the two interspecific hybrids, involving parthenogenesis in the latter and hybridogenesis in the former. Finally, the phyletic relationships among all Sicilian Bacillus taxa are summarized in a revised scheme, which also takes into account new evidence that B. atticus has contributed to hybrid constitution of the triploid B. lynceorum.     

New DAPI and FISH findings on egg maturation processes in related hybridogenetic and parthenogenetic Bacillus hybrids (Insecta, Phasmatodea)

Bacillus stick insects have proved adequate for studying a wide array of reproductive modes: sexual, parthenogenetic, hybridogenetic, androgenetic. Hybridogenetic strains (B. rossius-grandii) were thought to discard the paternal "grandii" haploset during first meiotic division and keep the "rossius" hemiclone, whereas the clonal B. whitei (=rossius/grandii) would maintain its hybrid structure by fusing back two nonsister nuclei-each derived from previously segregated heterospecific complements-by the end of the 2(nd) meiotic division. New investigations on laid eggs and ovariole squashes, either DAPI stained or FISH labeled, revealed that in hybridogens the "grandii" set is excluded from the germ line prior to meiosis and that a DNA extra-synthesis should occur to produce hemiclonal eggs after two cytologically normal meiotic divisions. On the other hand, in B. whitei eggs no genome segregation appears to occur and an intrameiotic DNA extra-synthesis must take place to produce 2n tetrachromatidic oocytes I; these divide twice and give unreduced clonal eggs. The new findings bring hybridogenetic oogenesis of Bacillus to be coincident with that of the known hemiclonal organisms and point to an independent onset of B. whitei from hemiclonal strains. In addition, B. whitei gains a closer resemblance to B. lynceorum owing to the sharing of a cytologically identical egg maturation mechanism, of the same maternal ancestor and of peculiar chromosomal features. It is here suggested that B. lynceorum originated from the incorporation of an "atticus" genome into a B. whitei egg, according to a pathway of repeated hybridization often occurred with other polyploid hybrids.     

Linkage between sexual and asexual lineages: genome evolution in Bacillus stick insects

The sexually reproducing stick insects Bacillus rossius and B. grandii are sharply differentiated in terms of allozyme gene alleles; B. atticus is a polyclonal automictic parthenogen sister to B. grandii grandii. Although well differentiated for coding genes, these hybridize to produce diploid (B. whitei=rossius/grandii) or triploid (B. lynceorum=rossius/grandii/atticus) clonal forms which reproduce apomictically. Allozyme analyses of unisexual Bacillus clearly establish their relationships from bisexual ancestor species as does the existence in all of them of several clones (especially in B. atticus) whose egg maturation allows regular recombination to occur. Bacillus taxa share the Bag320 satellite DNA family within different reproductive frameworks, allowing satellite variant homogenization to be uncoupled from fixation. The nested analysis of monomers reveals different patterns of sequence diversity: sexual reproduction includes both homogenization and variant fixation, whereas the slowing of molecular turnover processes and the absence of syngamy in the parthenogens realizes a similar range of sequence diversity at the level of the individual and supra‐individual, but with no fixation. On the other hand, the actual values of sequence diversity appear mostly linked to species traits – range size, copy number of repeats, number of hybrid crosses – and possibly transposon activity, rather than to the reproductive mode. In addition, the mitochondrial genome reveals a comparable level of cox2 sequence variability in sexual and parthenogenetic taxa, thus adding to clonal variability. From Bacillus and other stick insect complexes, an overall picture of genomic diversification of parthenogens is therefore beginning to emerge. To define those animals that reproduce by non‐canonical sexual modes (i.e. parthenogenesis, hybridogenesis), but make use of egg and meiotic mechanisms, the term meta‐sexual is proposed. © 2003 The Linnean Society of London. Biological Journal of the Linnean Society, 2003, 79 , 137–150.     

Reproductive features of homospecific hybridogenetically-derived stick insects suggest how unisexuals can evolve

Hybridogenetic reproduction has been demonstrated in both vertebrate and invertebrate unisexual hybrids. Its most peculiar feature is the transmission to the progeny of one invariant genome (hemiclone) through the egg and the replacement of the other by host fathering males. Bacillus hybridogens are the only known example of hemiclonal invertebrates; their comparison to Poeciliopsis and Rana systems helps in understanding peculiar and shared features of vertebrate and insect hybridogenesis. In P. monacha-lucida, the experimental production of non-hybrid progeny through the reunion of the maternal hemiclone with a homospecific paternal genome provided by males of the maternal ancestor leads to inviable or severely impaired sterile specimens, whereas in Rana esculenta viable offspring are the rule. The comparable synthetic B. rossius progeny (Rr) embodying the maternal R hemiclone and a paternal r haploset, appear perfectly viable and fertile, clearly demonstrating compatibility between the two homospecific genomes, and also supporting a lack of deterioration of the R hemiclone. This condition can be ascribed to the recent origin of the hemiclones, and also to the absence of lethal recessives, owing to their most likely derivation from an automictic doubling in the parthenogenetic mechanisms of the maternal ancestor. However, the hybridogenetic system breaks down in the gamete production of the majority of Rr females, since normal allele segregation also occurs in their progeny. These reproductive modes suggest a likely evolutionary dynamic for newly originated hybridogens: to achieve stability, an interruption of reproductive interactions with the maternal ancestor seems necessary. In stick insects, this constraint appears to be fulfilled in both areas of sympatry. The microevolutionary pathway suggested by the ecological scenario also supports the possibility that a shift of hemiclonal stick insect strains to clonality has occurred.     

The mitochondrial cytochrome oxidase II gene in Bacillus stick insects: ancestry of hybrids, androgenesis, and phylogenetic relationships

Sequencing of a cytochrome oxidase II (COII) gene fragment in Bacillus taxa provided evidence that the bisexual B. rossius is the maternal ancestor of the hybridogenetic B. rossius-grandii strains and revealed the same ancestry for both parthenogenetic hybrids: the diploid B. whitei (B. rossius/grandii grandii) and the triploid B. lynceorum (B. rossius/grandii grandii/atticus). Present data clearly demonstrate that all Bacillus unisexuals arose through asymmetrical hybridization events and realized a paraphyletic derivation from the B. rossius redtenbacheri subspecies. The invention of B. rossius mitochondrial DNA haplotypes in specimens with B. grandii grandii nuclear genomes revealed the occurrence of androgenesis in nature. Natural androgens represent a peculiar escape from hybridity and can help maintain the hybridogenetic system through the production of the fathering taxon via hybrid females. Results from the COII gene support the phyletic relationships among taxa suggested by previous taxonomical approaches, but also indicate a departure of B. grandii subspecies from the established taxonomy. Assuming the existence of a molecular clock, the evaluated substitution rate brings the splitting between B. rossius and B. grandii/B. atticus back to 22.79 +/- 2.65 myr before present, while the origin of hybrids appears to be much more recent (1.06 +/- 0.53 myr).     

Genome exclusion and gametic DAPI-DNA content in the hybridogenetic Bacillus rossius-grandii benazzii complex (Insecta Phasmatodea).

Among Sicilian stick insects, two hybridogenetic complexes have been discovered: Bacillus rossius-grandii benazzii and B. rossius-grandii grandii, which also produce androgenetic offspring. The egg maturation of the former is analyzed here through DAPI fluorometry, which, besides the assessment of the meiotic stages, also allows their DNA measurements and the analysis of sperm-head evolution into male pronuclei in these polyspermic eggs. Hybridogenetic eggs undergo an extrasynthesis of chromosomes, because two groups of n autobivalents (4C each) are segregated at metaphase 1st; the two groups must correspond to the pure parental species haplosets. Then the grandii chromosomes degenerate (1st polar body), while the rossius chromosomes divide further to produce two groups of n autodiads (2C each); one of them degenerates (2nd polar body), and the other is ready to perform syngamy (female pronucleus). Meanwhile, several B. grandii sperm evolve into male pronuclei by doubling their DNA (from 1C to 2C content) and assuming an interphase nucleus appearance. If regular mixis occurs, the F1 hybrid constitution is restored but, if it fails, a fusion between two sperms may occur, originating fully paternal descendants (natural androgenesis). The genome exclusion mechanism of stick-insect hybridogens appears to be more primitive than those observed in the already known hybridogenetic complexes of Poeciliopsis and Rana esculenta. Unfertilized eggs of hybridogens are capable of self-activation, but the cytology of the related clonally reproducing B. whitei indicates that its parthenogenetic mechanism stems from the hybridization event (hybrid theory) rather than from tychoparthenogenetic potentialities (spontaneous theory).     

Developmental instability, hybridization and heterozygosity in stick insects of the genus Bacillus (Insecta; Phasmatodea) with different modes of reproduction

Several genetic factors are assumed to influence developmental instability (DI). One is the level of heterozygosity, with higher levels often being associated with decreased DI; another is genetic incompatibility in hybrids, which in several cases has been shown to increase DI. The genus Bacillus includes species which have both amphigonic heterozygous reproducing populations and homozygous parthenogenetic reproducing populations (B. rossius rossius and B. r. redtenbacheri). Furthermore, Bacillus includes hybrid parthenogenetic species, which have very high levels of almost fixed heterozygosities (B. atticus, B. whitei, B. lynceorum). We investigated the phenotypic variance (σ²p) and the impact of hybridization and level of heterozygosity on DI in females from these populations and species of Bacillus. DI was estimated as fluctuating asymmetry (FA) for three bilateral traits: the labial palpus, the maxillary palpus and the antenna. For the labial palpus and maxillary palpus we found, in general, a lower level of DI in the amphigonic females compared with parthenogenetic counterparts from the same species and with parthenogenetic females from the three hybrid species. A higher DI of the antenna was found in the hybrid species when compared with both parthenogenetic and amphigonic populations of the nonhybrid species, suggesting that the genes controlling antenna development are located on the sex chromosomes. The development of the investigated bilateral characters in the hybrid species seemed to be affected more by factors relating to genetic incompatibilities as a consequence of hybridization than by the stabilizing force of increased heterozygosity. Only few differences in σ²p were observed, supporting the possibility that the observed differences in DI are related mainly to internal genetic factors. © 2006 The Linnean Society of London, Biological Journal of the Linnean Society, 2006, 87 , 249–259.     

Satellite sequence turnover in parthenogenetic systems: the apomictic triploid hybrid Bacillus lynceorum (Insecta, Phasmatodea)

In the genus Bacillus (Insecta, Phasmatodea) the Bag320 satellite DNA family is present in the bisexual B. grandii and in the related automictic nonhybrid B. atticus; it is lacking in the other bisexual taxon of the genus, B. rossius. This family of highly repeated sequences was analyzed for 11 populations of the apomictic triploid hybrid B. lynceorum. In the neighbor-joining dendrogram, B. lynceorum nucleotide sequences distribute, regardless of geographical origin, among two clusters, one also including all clones of the three B. atticus races, and the other including sequences of the B. grandii grandii subspecies. Thus, B. lynceorum is a trihybrid taxon: as the molecular approach definitively demonstrates, it embodies one haploid complement each of both B. grandii grandii and B. atticus, which must be added to that of B. rossius. The contribution of the latter species has already been assessed on karyological and allozymic grounds. A statistical analysis performed on p-distances shows that for the parental taxa, nucleotide substitution values are of comparable magnitudes at the population level but differ at the subspecific level, being higher for the bisexual taxon. In the apomictic hybrid, atticus- and grandii grandii- like sequences coexist with significantly different p-distance values. For three clones, the nucleotide compositions at the diagnostic loci suggest that gene conversion can occur between atticus- and grandii grandii-like monomers. On the whole, this supports bisexuality as a driving force in variant fixation and suggests that in Bacillus, different gametogenetic processes and different origins of the unisexuals are mirrored in genomic turnover rates of satellite DNA.      

Hybridogenesis through thelytokous parthenogenesis in two Cataglyphis desert ants

Hybridogenesis is a sexual reproductive system, whereby parents from different genetic origin hybridize. Both the maternal and paternal genomes are expressed in somatic tissues, but the paternal genome is systematically excluded from the germ line, which is therefore purely maternal. Recently, a unique case of hybridogenesis at a social level was reported in the desert ant Cataglyphis hispanica. All workers are sexually produced hybridogens, whereas sexual forms (new queens and males) are produced by queens through parthenogenesis. Thus, only maternal genes are perpetuated across generations. Here, we show that such an unusual reproductive strategy also evolved in two other species of Cataglyphis belonging to the same phylogenetic group, Cataglyphis velox and Cataglyphis mauritanica. In both species, queens mate exclusively with males originating from a different genetic lineage than their own to produce hybrid workers, while they use parthenogenesis to produce the male and female reproductive castes. In contrast to single‐queen colonies of C. hispanica, colonies of C. velox and C. mauritanica are headed by several queens. Most queens within colonies share the same multilocus genotype and never transmit their mates' alleles to the reproductive castes. Social hybridogenesis in the desert ants has direct consequences on the genetic variability of populations and on caste determination. We also discuss the maintenance of this reproductive strategy within the genus Cataglyphis.     

Unisexuality and Molecular Drive: Bag320 Sequence Diversity in <Emphasis Type="Italic">Bacillus</Emphasis> Taxa (Insecta Phasmatodea)

Satellite DNA variability follows a pattern of concerted evolution through homogenization of new variants by genomic turnover mechanisms and variant fixation by chromosome redistribution into new combinations with the sexual process. Bacillus taxa share the same Bag320 satellite family and their reproduction ranges from strict bisexuality (B. grandii) to automictic (B. atticus) and apomictic (B. whitei = rossius/ grandii; B. lynceorum = rossius/grandii/atticus) unisexuality. Thelytokous reproduction clearly allows uncoupling of homogenization from fixation. Both trends and absolute values of satellite variability were analyzed in all Bacillus taxa but B. rossius, on 906 sequenced monomers at all level of comparisons: intraspecimen, intrapopulation, interpopulation, intersubspecies, and interspecies. For unisexuals, allozymic and mitochondrial clones were also taken into account. Different reproductive modes (sexual/parthenogenetic) appear to explain observed variability trends, supporting Dover's hypothesis of sexuality acting as a driving force in the fixation of sequence variants, but the present analyses also highlight current spreading of new variants in B. grandii maretimi specimens and point to a biased sequence inheritance at the time of hybrid onset in the apomictic hybrids B. whitei and B. lynceorum. Evidence of biased gene conversion events suggests that, given enough time, sequence homogenization can take place in a unisexual such as B. lynceorum. On the contrary, the absolute values of sequence diversity in each taxon are linked to the species' range, time of divergence, and repeat copy number and, possibly, to transposon features. Satellite dynamics appears therefore to be the outcome of both general molecular processes and specific organismal traits.     

Identification of hemiclonal reproduction in three species of Hexagrammos marine reef fishes

Natural hybrids between the boreal species Hexagrammos octogrammus and two temperate species Hexagrammos agrammus and Hexagrammos otakii were observed frequently in southern Hokkaido, Japan. Previous studies revealed that H. octogrammus is a maternal ancestor of both hybrids; the hybrids are all fertile females and they frequently breed with paternal species. Although such rampant hybridization occurs, species boundaries have been maintained in the hybrid zone. Possible explanations for the absence of introgressions, despite the frequent backcrossing, might include clonal reproduction: parthenogenesis, gynogenesis and hybridogenesis. The natural hybrids produced haploid eggs that contained only the H. octogrammus genome (maternal ancestor) with discarded paternal genome and generated F₁‐hybrid type offspring by fertilization with the haploid sperm of H. agrammus or H. otakii (paternal ancestor). This reproductive mode was found in an artificial backcross hybrid between the natural hybrid and a male of the paternal ancestor. These findings indicate that the natural hybrids adopt hybridogenesis with high possibility and produce successive generations through hybridogenesis by backcrossing with the paternal ancestor. These hybrids of Hexagrammos represent the first hybridogenetic system found from marine fishes that widely inhabit the North Pacific Ocean. In contrast with other hybridogenetic systems, these Hexagrammos hybrids coexist with all three ancestral species in the hybrid zone. The coexistence mechanism is also discussed.     

Obligatory parthenogenesis and TE load: Bacillus stick insects and the R2 non‐LTR retrotransposon

Transposable elements (TEs) are selfish genetic elements whose self‐replication is contrasted by the host genome. In this context, host reproductive strategies are predicted to impact on both TEs load and activity. The presence and insertion distribution of the non‐LTR retrotransposon R2 was here studied in populations of the strictly bisexual Bacillus grandii maretimi and of the obligatory parthenogenetic Bacillus atticus atticus. Furthermore, data were also obtained from the offspring of selected B. a. atticus females. At the population level, the gonochoric B. g. maretimi showed a significantly higher R2 load than the obligatory parthenogenetic B. a. atticus. The comparison with bisexual and unisexual Bacillus rossius populations showed that their values were higher than those recorded for B. a. atticus and similar, or even higher, than those of B. g. maretimi. Consistently, an R2 load reduction is scored in B. a. atticus offspring even if with a great variance. On the whole, data here produced indicate that in the obligatory unisexual B. a. atticus R2 is active and that mechanisms of molecular turnover are effective. Furthermore, progeny analyses show that, at variance of the facultative parthenogenetic B. rossius, the R2 activity is held at a lower rate. Modeling parental‐offspring inheritance, suggests that in B. a. atticus recombination plays a major role in eliminating insertions rather than selection, as previously suggested for unisexual B. rossius progeny, even if in both cases a high variance is observed. In addition to this, mechanisms of R2 silencing or chances of clonal selection cannot be ruled out.     

Allozymic characterization of Sardinian Bacillus rossius (Rossi) and B. atticus Brunner (Insecta Phasmatodea)

The allozymic characterization of newly discovered populations of B. atticus and B. rossius is given. Most Sardinian B. rossius demes belong to the subspecies B. r. rossius, Southeastern populations are B. r. redtenbacheri, while the central ones appear to be intermediate. Like all Italian B. atticus, also Sardinian demes belong to B. a. caprai. The presence of B. atticus alleles in syntopic B. rossius populations is explained through gene introgression due to past hybridization events between the two taxa.     

Cytogenetic studies on Bacillus grandii grandii and B. grandii benazzii (Insecta,Phasmatodea): karyotype description,constitutive heterochromatin and nucleolus organizer regions

The karyotype of the new Sicilian subspecies Bacillus grandii benazzii is described and compared to that of B. g. grandii. Both taxa share constant, very similar chromosome sets with 2n=34, XX, female and 2n=33, XO, male, the sex chromosomes being the third element of the complement. C-banding reveals that a centromeric heterochromatin mass is present on all chromosomes, but no interstitial bands are observed. Differences between karyotypes reside in the satellites which are located on pairs 13 and 16 in B. g. benazzii and only on pair 16 in B. g. grandii, in contracted metaphases larger satellites are positively C-banded. Furthermore, Ag-detected NORs are seen on pair 13 in B. g. benazzii and on pair 16 in B. g. grandii. Relationships between constitutive heterochromatin and NORs, NOR heteromorphism and the position of the 34-chromosome taxa within the genus are discussed.     

Characterization of microsatellite loci in the stick insects Bacillus rossius rossius,Bacillus rossius redtenbacheri and Bacillus whitei (Insecta: Phasmatodea)

Five microsatellite markers were obtained from a dinucleotide enriched genomic library of the stick insect Bacillus rossius rossius. The markers were tested in three species of Bacillus. All loci were polymorphic when tested across species. The number of alleles at each locus was low (maximum four alleles), but different allelic patters were observed among the species.     

Unisexual hybrids break through an evolutionary dead end by two-way backcrossing

Unisexual vertebrates (i.e., those produced through clonal or hemiclonal reproduction) are typically incapable of purging deleterious mutations, and, as a result, are considered short-lived in evolutionary terms. In hemiclonal reproduction (hybridogenesis), one parental genome is eliminated during oogenesis, producing haploid eggs containing the genome of a single parent. Hemiclonal hybrids are usually produced by backcrossing hemiclonal hybrids with males of the paternal species. When hemiclonal hybrids from a genus of greenlings (Hexagrammos) are crossed with males of the maternal species, the progeny are phenotypically similar to the maternal species and produce recombinant gametes by regular meiosis. The present study was conducted to determine if the hemiclonal genome is returned to the gene pool of the maternal species in the wild. Using a specific cytogenetic marker to discriminate between such progeny and the maternal species, we observed that Hexagrammos hybrids mated with maternal and paternal ancestors at the same frequency. This two-way backcrossing in which clonal genomes are returned to the gene pool where they can undergo recombination plays an important role in increasing the genetic variability of the hemiclonal genome and reducing the extinction risk. In this way, hybrid lineages may have survived longer than predicted through occasional recombinant generation.     

Comparative vitellin pattern and cladogenesis in Bacillus (insecta: Phasmatodea)

• 1.1. Under denaturing conditions (SDS-PAGE) the two natural vitellins of Bacillus taxa released five different polypeptides (A₁, A₂, A₃, B₁, B₂).
• 2.2. A₂ and B₂ bands from the two bisexual species (B. rossius and B. grandii) were found to differ; furthermore a non-vitellin yolk protein characterizes the subsepecies B.g. benazzii.
• 3.3. From gels and their densitometric scanning profiles it is clear that parental polypeptides are expressed in the thelytokous parthenogenetic hybrids (B. whitei, B. lynceorum) and in the hybridogenetic B. rossius-grandii benazzii.
• 4.4. A comparative approach of vitellin patterns appears fully adequate for tracing phylogenetic relationships and recognizing cladogenetic events.

     

Repeated evolution of queen parthenogenesis and social hybridogenesis in Cataglyphis desert ants

Over the last decade, genetic studies on social insects have revealed a remarkable diversity of unusual reproductive strategies, such as male clonality, female clonality, and social hybridogenesis. In this context, Cataglyphis desert ants are useful models because of their unique reproductive systems. In several species, queens conditionally use sexual reproduction and parthenogenesis to produce sterile workers and reproductive queens, respectively. In social hybridogenesis, two distinct genetic lineages coexist within a population, and workers result from mating between partners of different lineages; in contrast, queens and males are both produced asexually by parthenogenesis. Consequently, nonreproductive workers are all interlineage hybrids, whereas reproductives are all pure lineage individuals. Here, we characterized the reproductive systems of 11 species to investigate the distribution of the conditional use of sex and social hybridogenesis in Cataglyphis. We identified one new case in which sexual reproduction was conditionally used in the absence of dependent‐lineage reproduction. We also discovered five new instances of social hybridogenesis. Based on our phylogenetic analyses, we inferred that both the conditional use of sex and social hybridogenesis independently evolved multiple times in the genus Cataglyphis.     

Large-scale distribution of hybridogenetic lineages in a Spanish desert ant

Recently, a unique case of hybridogenesis at a social level was reported in local populations of the desert ants Cataglyphis. Queens mate with males originating from a different genetic lineage than their own to produce hybrid workers, but they use parthenogenesis for the production of reproductive offspring (males and females). As a result, non-reproductive workers are all inter-lineage hybrids, whereas the sexual line is purely maternal. Here, we show that this unorthodox reproductive system occurs in all populations of the ant Cataglyphis hispanica. Remarkably, workers are hybrids of the same two genetic lineages along a 400 km transect crossing the whole distribution range of the species. These results indicate that social hybridogenesis in C. hispanica allows their maintenance over time and across a large geographical scale of two highly divergent genetic lineages, despite their constant hybridization. The widespread distribution of social hybridogenesis in C. hispanica supports that this reproductive strategy has been evolutionarily conserved over a long period.