Male-killing Wolbachia in a flour beetle

The bacteria in the genus Wolbachia are cytoplasmically inherited symbionts of arthropods. Infection often causes profound changes in host reproduction, enhancing bacterial transmission and spread in a population. The reproductive alterations known to result from Wolbachia infection include cytoplasmic incompatibility (CI), parthenogenesis, feminization of genetic males, fecundity enhancement, male killing and, perhaps, lethality Here, we report male killing in a third insect, the black flour beetle Tribolium madens, based on highly female-biased sex ratios of progeny from females infected with Wolbachia. The bias is cytoplasmic in nature as shown by repeated backcrossing of infected females with males of a naturally uninfected strain. Infection also lowers the egg hatch rates significantly to approximately half of those observed for uninfected females. Treatment of the host with antibiotics eliminated infection, reverted the sex ratio to unbiased levels and increased the percentage hatch. Typically Wolbachia infection is transmitted from mother to progeny, regardless of the sex of the progeny; however, infected T. madens males are never found. Virgin females are sterile, suggesting that the sex-ratio distortion in T. madens results from embryonic male killing rather than parthenogenesis. Based on DNA sequence data, the male-killing strain of Wolbachia in T. madens was indistinguishable from the CI-inducing Wolbachia in Tribolium confusum, a closely related beetle. Our findings suggest that host symbiont interaction effects may play an important role in the induction of Wolbachia reproductive phenotypes.     

Sex-ratio drive in Drosophila simulans: variation in segregation ratio of X chromosomes from a natural population

The sex-ratio trait that exists in a dozen Drosophila species is a case of naturally occurring X chromosome drive that causes males to produce female-biased progeny. Autosomal and Y polymorphism for suppressors are known to cause variation in drive expression, but the X chromosome polymorphism has never been thoroughly investigated. We characterized 41 X chromosomes from a natural population of Drosophila simulans that had been transferred to a suppressor-free genetic background. We found two clear-cut groups of chromosomes, sex-ratio and standard. The sex-ratio X chromosomes differed in their segregation ratio (81-96% females in the progeny), the less powerful drivers being less stable in their expression. A sib analysis, using a moderate driver, indicated that within-X variation in drive expression depended on genetic (autosomal) or epigenetic factors and that the age of the males also affected the trait. The other X chromosomes produced equal or roughly equal sex ratios, but again with significant variation. The continuous pattern of variation observed within both groups suggested that, in addition to a major sex-ratio gene, many X-linked loci of small effect modify the segregation ratio of this chromosome and are maintained in a polymorphic state. This was also supported by the frequency distribution of sex ratios produced by recombinant X chromosomes.     

Impact of embryo technologies on secondary sex ratio in rabbit

Increasing evidence indicates that assisted reproductive technologies (ARTs) disturb skewed sex-ratio and induce sex-dimorphic postnatal effects. Undoubtedly, the combination of multiple ovulation and embryo transfer (MOET) together with the use of vitrification technique (MOVET) is currently being used in breeding programs. However, since the first case of sex skewing reported in 1991, the accumulative and long-term transmission of skewed sex-ratio to future generations has not been thoroughly evaluated. Here we test as MOVET program induce a skewed sex ratio, and we consider skewed sex ratio transmission to future generations. To this end, we first evaluated the F1 generation, demonstrating that a MOVET program causes a severe imbalance skewed secondary sex ratio (SSR) towards male by 12%. This imbalanced persist after a second MOVET program (F2 generation), with an accumulative skewed SSR towards male by 25%. Finally, using a crossbred generation derived from crossing F1 males derived from a MOVET program with naturally-conceived (NC) females, we show that the imbalance skewed SRR persist. Bodyweight comparison between MOVET animals and NC counterparts revealed significant changes at birth, weaning and adulthood. However, there was a significant interaction between F2 MOVET animals and sex, demonstrating an apparent accumulative sex-dimorphic effect. At adulthood, MOVET derived males presented a lower body weight. In conclusion, we show that the MOVET program causes a direct, accumulative and long-term transmission of skewed SSR.     

Association between loci with deleterious alleles and distorted sex ratios in an inbred line of tilapia (Oreochromis aureus)

Three microsatellite markers (UNH159, UNH231, and UNH216) were examined for association with both deleterious genes and sex-ratio distortions in a full-sib family of 222 progeny from the fourth generation of a meiogynogenetic tilapia line (Oreochromis aureus). The three markers were mapped previously to different linkage groups and were shown to be associated with genes with deleterious alleles in this line. A restricted maximum likelihood model was used for analysis of major effects and their interactions on sex ratio and viability. This model was based on selective mortality of genders, ignoring effects of possible sex-determining genes. The results showed that deleterious genes linked to UNH216 and UNH231 exert higher lethality in females than in males (P < .0005 and P < .05, respectively). UNH159 was not associated directly with sex ratio distortion, but acts strongly as a modifier of sex ratio in combination with UNH216 and UNH231. Each of the three loci was found to have a significant effect on viability (P < .05) in the maximum likelihood analysis. The deleterious single-locus effects act strongly against females, while most of the epistatic interactions exert higher lethality in males. This contradiction results in a close to 1:1 sex ratio at maturity. The genetic mechanism and significance of such a balance between genders are still unknown. A detailed analysis of sex-specific lethality may be applied by screening in appropriate series of matings and fine mapping with additional markers. Our data suggest that UNH216 and UNH231 are linked to sex ratio distortion genes and that UNH159 may be linked to a modifier of these genes.     

Nestling sex ratios in a population of Bluethroats Luscinia svecica inferred from AFLP™ analysis

We studied the sex ratio of Bluethroat Luscinia svecica broods using AFLPs. Our aim was to test whether there is a bias towards males that could be explained by sexual selection theories, or conversely, a bias towards females that could help explain the female-biased sex ratio among juveniles observed at a wintering site. The AFLP technique was reliable in sexing the nestlings from even small initial DNA quantities. Given the large number of polymorphic markers that can be obtained for each primer combination, the probability of detecting a W-chromosome-linked fragment is reasonably high. As a consequence, this method could be used in other species for sex-ratio studies and for other genetic purposes. Among 246 nestlings, we found an overall proportion of males of 50.8% at hatching and the sex-ratio variation using broods as independent units was not significantly different from expectation under a binomial distribution. None of the parental and environmental variables tested changed significantly the deviance to the model. Thus, sex determination in the Bluethroat seems to match the classical Mendelian model of a 1:1 sex ratio and cannot explain the biased sex ratio towards juvenile females found at the wintering site.     

Meiotic drive alters sperm competitive ability in stalk-eyed flies.

Meiotic drive results when sperm carrying a driving chromosome preferentially survive development. Meiotic drive should therefore influence sperm competition because drive males produce fewer sperm than non-drive males. Whether meiotic drive also influences the competitive ability of sperm after ejaculation is unknown. Here we report the results from reciprocal crosses that are designed for estimating the sperm precedence of male stalk-eyed flies (Cyrtodiopsis whitei) with or without X-linked meiotic drive. We find that nearly half of all sex-ratio males, as compared with 14% of non-sex-ratio males, fail to produce young in a reciprocal cross. Furthermore, the proportion of progeny sired by a sex-ratio male in a female jointly inseminated by a non-sex-ratio male was less than expected from the number of sperm transferred. These effects are not due to differential sperm storage by females because, after a single mating with a sex-ratio male, all females stored sperm and because two sex-ratio males share paternity after jointly mating with a female. In addition to demonstrating a new mechanism of sperm competition, these results provide insight into the maintenance of sex-ratio polymorphisms. Sex-ratio males have less than one-half the fertility of non-sex-ratio males, as is required in order for frequency-dependent selection on males to produce a stable sex-ratio polymorphism.     

The genetic regulation of liver microsomal CYP2E1 activity among strains of the viviparous fish Poeciliopsis

CYP2E1 expression was examined within, among, and in F(1) and backcross progeny of strains (P. monacha S68-5; P. viriosa M65-23) of the viviparous fish Poeciliopsis. CYP 2E1 activity varied dramatically in P. monacha, and P. viriosa (3.9+/-0.8 and 9.6+/-1.3 microg/min/mg) as well as the temperature which gave maximal activity (T(0)=25 degrees C and 31 degrees C). F(1) individuals from a crosses between P. monacha and P. viriosa, produced progeny whose CYP2E1 activity segregated into three different groups: (1) phenotypically the same as P. viriosa; (2) intermediate between the two parental strains; and (3) phenotypically the same as P. monacha. When a male P. monacha was crossed with a female P. viriosa 25% of the offspring had an intermediate phenotype and 65% the maternal P. viriosa phenotype. From the same cross, 85% of the females progeny had the maternal phenotype, while 80% of male progeny had the intermediate and paternal phenotype, suggesting an effect of the maternal genome on the F(1) phenotype. Among F(1) fish the T(0) was evenly distributed between parental values. In the backcross of a F(1) female to a male P. viriosa, CZX-6-hydroxylase activity segregated into the same three phenotypes with 60% of the progeny expressing the P. monacha phenotype. From the same cross, 70% of females and 40% of males expressed the P. monacha phenotype. The T(0) in the backcross were evenly distributed between the two parental values and the sex ratio among progeny was different than expected.     

Hierarchical selection theory and sex ratios I. General solutions for structured populations

Models of sex-ratio evolution in structured populations are derived with G.R. Price's covariance form for the hierarchical analysis of natural selection (1970, Nature 227, 520-521). Previous work on competition among related males for mates (local mate competition), competition among related females for a limiting resource (local resource competition), inbreeding, group selection, and asymmetry of genetic inheritance between males and females, are subsumed under a general formulation for sex-ratio biases in structured populations. I found that the evolutionarily stable strategy sex ratio (males:females) for diploids is 1 - rho m:1 - rho f, where rho m is the regression coefficient of relatedness of the controlling genotypes on males competing for mates, rho f is the regression of controlling genotypes on females that compete for a fixed, limiting resource, and there is no inbreeding. For inbreeding and no competition among females, the evolutionarily stable strategy is 1 - rho m:1 + rho mf, where rho mf is the regression of controlling genotypes on females' mates.     

Effect of mating and parasitism regimes on progeny production and sex-ratio of Campoletis chlorideae Uchida

The ichneumonid parasitoid, C. chlorideae is an important natural enemy of pod borer/bollworm, Helicoverpa armigera (Hubner) in different agro-ecosystems. The sex-ratio of parasitoids has an important bearing on the population build up of the natural enemies for biological control of insect pests. Therefore, the present studies were conducted to gain an understanding of the influence of mating behaviour and abundance of the insect host on fecundity and sex-ratio of the parasitoid, C. chlorideae. There was no significant influence of number of matings and abundance of the insect host on cocoon formation, adult emergence, and larval and pupal periods of C. chlorideae. However, fecundity and female longevity were significantly influenced by mating and abundance of the insect host. There was a significant and positive correlation (r = 0.84**) between longevity and fecundity of C. chlorideae females. The unmated C. chlorideae females produced only males. Nearly 20% of the females that had mated twice were able to parasitize the H. armigera larvae successfully. The sex-ratio of the progeny from females that had mated twice was male biased. Females mated with males from the unmated females produced significantly less numbers of females than those mated with males from the fertilized females, indicating genetic regulation of sex-ratio in C. chlorideae.     

Fitness effects of a selfish gene (the Mus t complex) are revealed in an ecological context

In wild house mice, genes linked to the t transmission distortion complex cause meiotic drive by sabotaging wild-type gametes. The t complex is consequently inherited at frequencies higher than 90%. Yet, for unclear reasons, in wild mouse populations this selfish DNA is found at frequencies much lower than expected. Here, we examine selection on the t complex in 10 seminatural populations of wild mice based on data from 234 founders and nearly 2000 progeny. Eight of the 10 populations decreased in t frequency over one generation, and the overall frequency of t haplotypes across all 10 populations was 48.5% below expectations based on transmission distortion and 34.3% below Mendelian (or Hardy-Weinberg) expectations. Behavioral and reproductive data were collected for 10 months for each population, and microsatellite genotyping was performed on seven of the populations to determine parentage. These combined data show t-associated fitness declines in both males and females. This is the first study to show evidence for a reduction in the ability of +/t males to maintain territories. Because females tend to mate with dominant males, impairment of territorial success can explain much of the selection against t observed in our populations. In nature, selection against heterozygote carriers of the t complex helps solve the puzzlingly low t frequencies found in wild populations. This ecological approach for determining fitness consequences of genetic variants has broad application for the discovery of gene function in general.     

Intralocus sexual conflict,adaptive sex allocation,and the heritability of fitness

Intralocus sexual conflict arises when selection favours alternative fitness optima in males and females. Unresolved conflict can create negative between‐sex genetic correlations for fitness, such that high‐fitness parents produce high‐fitness progeny of their same sex, but low‐fitness progeny of the opposite sex. This cost of sexual conflict could be mitigated if high‐fitness parents bias sex allocation to produce more offspring of their same sex. Previous studies of the brown anole lizard (Anolis sagrei) show that viability selection on body size is sexually antagonistic, favouring large males and smaller females. However, sexual conflict over body size may be partially mitigated by adaptive sex allocation: large males sire more sons than daughters, whereas small males sire more daughters than sons. We explored the evolutionary implications of these phenomena by assessing the additive genetic (co)variance of fitness within and between sexes in a wild population. We measured two components of fitness: viability of adults over the breeding season, and the number of their progeny that survived to sexual maturity, which includes components of parental reproductive success and offspring viability (RS^V). Viability of parents was not correlated with adult viability of their sons or daughters. RS^V was positively correlated between sires and their offspring, but not between dams and their offspring. Neither component of fitness was significantly heritable, and neither exhibited negative between‐sex genetic correlations that would indicate unresolved sexual conflict. Rather, our results are more consistent with predictions regarding adaptive sex allocation in that, as the number of sons produced by a sire increased, the adult viability of his male progeny increased.     

Sex-ratio depression as a brood-pattern criterion of radiation damage inDrosophila melanogaster

The results of analyses of the brood variance in the relative frequency of females to males in the progeny ofD. melanogaster males having structurally-normal chromosomes, not only established that there is a brood-curve of sex-ratio depression in the progeny of irradiated males, but also revealed that there was a curve of sex-ratio change in successive broods of offspring from unirradiated males even though the ratio of females to males in the total progeny was 1 : 1. The brood curve of sex-ratio change in both series of control males tended to be bimodal, with an increasing frequency of female offspring during the first five or six days, followed by a progressive decrease until about the 12th day, than subsequently an increase until about the 15–16th days, with perhaps again a decrease in the latest (17–24d) broods. The bimodality of the curve appeared to be the consequence of a complex relationship between brood-size, brood-rank and sex-ratio change. Following irradiation of the males with 3000r of X-rays, the incidence of female offspring decreased linearly for the first eight days, with a low of 44.32% in the 6–8d broods. This was followed by an abrupt return to a 1 : 1 sex-ratio in the 9th day and subsequent (10–24d) broods. The recovery of 50% female offspring in the 9th-day brood was interpreted as supporting evidence for the thesis that this brood was derived from cells that had been spermatogonia at the time of irradiation of the parental males. Supported by Research Grant GM 15009-01 from the Public Health Service (U.S.).     

Sex determination in crayfish: are intersex Cherax quadricarinatus (Decapoda, Parastacidae) genetically females?

In the Australian red-claw crayfish Cherax quadricarinatus (von Martens) (Decapoda, Parastacidae), a gonochoristic species, seven different combinations of intersex individuals (with both male and female genital openings) have been described. However, to date, the genetic basis for this phenomenon has not been investigated. This study was designed to test a simple chromosome-based sex-determination model for C. quadricarinatus that assumes the male to be the homogametic (ZZ) sex. According to our model, intersex individuals that are functionally males are genetically females (WZ). Individual crosses were performed between intersex and female crayfish, with control crosses being performed between normal males and females. The control crosses yielded, in most cases, the expected 1:1 sex ratio in the F1 progeny. Crosses between intersex individuals and females yielded a 1:3 (male:female) sex ratio in most crosses. According to our hypothesis, one-third of the females produced in a cross of a female with an intersex animal should be WW females. The hypothesis was tested by crossing normal males with F1 females, which were progeny of intersex fathers. These crosses yielded almost 100% females, a finding that conforms to the above-suggested sex determination model for C. quadricarinatus and the female WZ genotype of intersex individuals.     

Identifizierung der genetischen Geschlechtschromosomen bei der monogenen Schmeißfliege Chrysomya rufifacies (Calliphoridae,Diptera)

Previous investigations have shown the sex determination in the monogenic blowfly Chrysomya rufifacies to be controlled by a cytologically not discernible homogamety-heterogamety mechanism in the female. Female-producing (thelygenic) females are assumed to be heterozygous for a dominant female sex realizer (F′) with sex-predetermining properties, while male-producing (arrhenogenic) females as well as males are supposed to be homozygous for the recessive allele (f). In order to identify the genetic sex chromosomes of C. rufifacies among its five pairs of long euchromatic chromosomes (nos.1–5) plus one pair of small heterochromatic ones (no. 6), all chromosomes were marked by reciprocal translocations induced by X-ray treatment of adult males. The inheritance of thirteen different heterozygous translocations has been analyzed. All of the translocations (eleven) between two of the four longer chromosomes did not show sex-linked inheritance, thus demonstrating the autosomal character of the chromosomes nos 1, 2, 3 and 4. The same is true for the translocation T6 (2/6). Therefore the small heterochromatic chromosome no. 6, corresponding to the morphologically differentiated sex chromosomes within the amphogenic calliphorid species, remains without sex determining function in the monogenic fly. This could be confirmed by the analysis of monosomic (monosomy-6) and trisomic (trisomy-6) individuals, which resulted from meiotic non-disjunction in T6/+ translocation heterozygotes. Contrary to these translocations, the heterozygous 5/2 translocation (T14) exhibited sex-linked inheritance: There was but a very low frequency (0,76 per cent) of recombinants resulting from crossing-over between F′/f and the translocation breakage point in thelygenic F₁ T14/+ females. The sex-linked inheritance of T14 was confirmed by the progeny of a thelygenic F₁ T14/+ female crossed to a homozygous T14/T14 translocation male. Among the offspring of that F₁ T14/+ female, which had received the translocation from its father, all of the F₂ T14/+ females were thelygenic compared to their arrhenogenic T14/T14 sisters. These results prove that the chromosomes of pair no. 5 genetically act as X′X-XX sex chromosomes in C. rufifacies.     

Effects of parent age at mating on reproductive response of Glyptapanteles flavicoxis (Hymenoptera: Braconidae), a larval parasitoid of the gypsy moth (Lepidoptera: Lymantriidae)

Glyptapanteles flavicoxis (Marsh) (Hymenoptera: Braconidae) is a gregarious larval parasitoid of the Indian gypsy moth Lymantria obfuscata (Walker) (Lepidoptera: Lymantriidae), that is believed to have potential for inundative releases against gypsy moth populations, because it can be reared in large numbers with few hosts. Unfortunately, sex ratios in laboratory reared G. flavicoxis are usually male-biased, hindering efforts to mass release this species for biological control by making the production of females costly. Because parental age at time of mating is known to affect the sex ratio in some Braconidae, we crossed haploid males and virgin females at 0, 1, 4, 9, and 16 d old with at least 10 trials for each of the 25 combinations. Numbers and sex ratios of progeny produced by females each day were recorded. Both progeny and sex ratios (percentage of females) among progeny produced by ovipositing females of G. flavicoxis decreased markedly over time, so only the first days production need be used in mass rearing. The reduction in the proportion and numbers of females among progeny as females aged is consistent with sperm depletion. Approximately 30% of females in all age classes mated to newly emerged males (day 0) produced all male progeny, whereas only 10-15% of those mated to older males failed to produce any daughters. When crosses with only male progeny were excluded from the analysis, females mated to males 1 d old had higher sex ratios in progeny than those mated to males in other age classes. In addition, females mated the day that they emerged tended to have progeny with the highest sex ratios.     

Genetic analysis of fertility of male mice with various t-haplotypes

Fertility of 47 mouse males carrying various combinations of lethal, t-haplotypes (t6/tw18, t12/tw18, Tw73/tw12, tw5/tw18, t6/dt5, t12/tw12, tw5/twPa-1, tw18/twPa-1, tw5/tw12) was studied in crosses with females of different genotypes. The t-haplotypes studied belong to 7 main groups of complementation. The presence of at least two factors of fertility in the t-complex was revealed. The influence of female genotype on the degree of male fertility was also demonstrated. The data presented confirm that different combinations of lethal complete t-haplotypes exhibit sterility, with the exception of t8/tw18 compound.     

Genetic analysis of complementation of t-haplotypes in mice

Complementation for viability of the compounds tx/ty in complete lethal haplotypes t6, t12, tw5, tw12, tw18, tw73, twPa-1 belonging to the seven basic complementation groups was studied. In the overwhelming majority of crosses a lack of tx/ty offspring was revealed but there is also evidence of non-complete complementation of the tx/ty haplotypes, except for T/tw73XT/tw12, T/tw18XT/t12, T/tw73XT/tw18 crosses in which the level of complementation for viability exceeds 100%. The maternal effect for compound variability was significantly revealed in 9 out of the 19 matings. In addition, reciprocal matings of heterozygotes T/txXT/ty were tested for fertility. In two cases, the maternal effect was also revealed for this character. Comparison of data on viability of the compounds and the fertility of the original strains gives grounds to believe that the tx/ty haplotypes are not completely recessive and partially manifest themselves in the heterozygotes, decreasing fertility in a number of matings. In some cases, these t-haplotypes yield an effect, close to negative complementation, which may be associated with the interaction of the proteins of t-haplotypes.     

Analysis of Autosomal Polygenic Variation for the Expression of Haldane''s Rule in Flour Beetles

Haldane's rule states that, in interspecific crosses, when hybrid viability or fertility is diminished more in one sex of the hybrids than in the other, the heterogametic sex is more adversely affected. We used quantitative genetic methods to investigate the genetic basis of variation for the expression of the viability aspect of Haldane's rule when Tribolium castaneum males are crossed to Tribolium freemani females. Using a half-sib design, we found significant genetic variance for the expression of Haldane's rule, i.e, variation among T. castaneum sires in the hybrid sex ratios produced by their sons. We also derived 23 independent lineages from the same base population by 8 generations of brother-sister mating. From the same experiments, we also found heritable variation among surviving hybrid males in the incidence of antennal deformities. Upon inbreeding, the variance of both traits (hybrid sex ratio and proportion deformities) increased substantially but the means changed little. Because fitness within T. castaneum lineages declined substantially with inbreeding, we infer that hybrid male viability may have a different genetic basis than viability fitness within species. Deleterious recessive alleles held within species by mutation/selection balance appear not to be a major contributor to hybrid incompatibility.     

Genetics of mother-dependent sex ratio in blue mussels (Mytilus spp.) and implications for doubly uniparental inheritance of mitochondrial DNA

Previous studies have shown that in most pair matings of Mytilus edulis, M. trossulus, and M. galloprovincialis there is a large sex-ratio bias in favor of either males or females. The degree of bias is a characteristic property of the female parent, as matings of the same female with different males produce the same sex ratio, but matings of the same male with different females produce different sex ratios. All three species possess the unusual feature of doubly uniparental inheritance of mitochondrial DNA (mtDNA); i.e., they contain two distinct types of mtDNA, one that is transmitted matrilinearly and one that is transmitted patrilinearly. This coupling of sex and mtDNA transmission raises the possibility that the mechanism of sex-ratio determination in mussels might be under the control of the mtDNA of the female parent. Here we present data from pedigreed crosses that confirm the previous observations that in mussel matings there is a strong sex-ratio bias and that the bias is under the control of the female parent. In addition, these data strongly suggest that this control is exercised by the mother's nuclear rather than mitochondrial genotype. Making use of these findings we develop a model of mother-dependent sex determination and use data from crosses involving wild females to test the model's predictions at the population level.     

The Relative Influence of Sex of Progeny on the Lethal Expression of the Sonless Gene in DROSOPHILA MELANOGASTER

The sex-linked maternally influenced lethal gene, sonless (snl), in D. melanogaster was shown to affect the survival of daughters as well as sons of snl/snl mothers but to a lesser degree. Interaction studies of sonless with the sex altering mutants transformer (tra) and doublesex (dsx) revealed that any alteration toward increased masculinity of progeny from snl/snl females reduced their zygotic viability, independent of snl dosage or Y chromosome presence or absence. Possible implications of these results are discussed and the original gene action hypothesis for snl is extended to incorporate these new findings.-Sonless is non-allelic but closely linked to rudimentary (r) and fused (fu), two mutants with similar sex-ratio patterns.