Hybrid Lethal Systems in the Drosophila Melanogaster Species Complex. II. the Zygotic Hybrid Rescue (Zhr) Gene of D. Melanogaster

Hybrid females from Drosophila simulans females X Drosophila melanogaster males die as embryos while hybrid males from the reciprocal cross die as larvae. We have recovered a mutation in melanogaster that rescues the former hybrid females. It was located on the X chromosome at a position close to the centromere, and it was a zygotically acting gene, in contrast with mhr (maternal hybrid rescue) in simulans that rescues the same hybrids maternally. We named it Zhr (Zygotic hybrid rescue). The gene also rescues hybrid females from embryonic lethals in crosses of Drosophila mauritiana females X D. melanogaster males and of Drosophila sechellia females X D. melanogaster males. Independence of the hybrid embryonic lethality and the hybrid larval lethality suggested in a companion study was confirmed by employing two rescue genes, Zhr and Hmr (Hybrid male rescue), in doubly lethal hybrids. A model is proposed to explain the genetic mechanisms of hybrid lethalities as well as the evolutionary pathways.     

The Nature of Heterosis and Methods of Enhancing and Fixing It in a Series of Generations without Hybridization

The long-standing studies into the problem of heterosis on the silkworm and other objects are reviewed. Silkworms are divided by sex for hybridization using the genetic marking of the eggs by sex, an improved method of ameiotic parthenogenesis providing all-female progeny, or the method of obtaining all-male progeny as a result of the death of eggs with female embryos under the influence of two non-allele embryonic lethals balances in the Z-chromosome. The experimental data suggest that heterosis is determined by the heterozygosity of lethals and semilethals, rather than depending on the heterozygosity of all gene types. The number of modifier genes that control viability plays a big role in the vigor of heterosis. An effective method is proposed to enhance the vigor of heterosis through the selection of one or two parents of a hybrid for viability, both of which received a strong semilethal. In hybrids, this semilethal is suppressed by the normal allele, and then a more unbalanced complex of modifier genes initiates a heterosis two to three times more vigorous than in standard hybrids. A method is developed to fix heterosis through the backcrosses of females of the high-heterosis hybrids to the absolutely homozygous males of androgenetic origin obtained from these females. These crosses preserve the main genotype of the hybrid and eliminate lethals and semilethals, and, as a result, heterosis is not damped during standard intrahybrid reproduction.     

Analysis of Dysgenesis-Induced Lethal Mutations on the X Chromosome of a Q Strain of DROSOPHILA MELANOGASTER

The Q strain known as v6 was tested for its ability to induce X-linked lethal mutations in male and female hybrids from crosses with M strains in the P-M system of hybrid dysgenesis. All measurements of the mutation rate were made on the X chromosome derived from the v6 strain. The lethal rate for young hybrid males from the cross M female X v6 male was 1.11% per chromosome. For older males, it was only 0.44%, suggesting that there is less mutational or more repair activity in the germ cells of the older males or that mutant cells are selectively eliminated as the hybrid males age. The lethal rate for hybrid females from comparable crosses was approximately the same for both ages that were tested. However, it was substantially less than the rate for the hybrid males--only 0.26% per chromosome. Genetically identical hybrid females from reciprocal crosses also showed a low mutation rate, 0.13% per chromosome. Again, there was no difference between young and old flies. Mapping experiments established that most of the lethal mutations that were recovered from the male and female hybrids were located in two regions on the X chromosome, one between bands 14B13 and 15A9 , the other between bands 19A1 and 20A , which encompasses the maroonlike locus. More refined mapping of the lethals in the maroonlike region demonstrated that the vast majority of these affected a single gene located in band 19C4 . Cytological analysis of the lethal chromosomes revealed that several carried rearrangements, including inversions, duplications and deficiencies. Chromosome breakage occurred primarily in bands 14D1 -3 and 18F- 20A , and most of the breaks in the latter segment were located in 19C . However, rearrangements involving 19C and mutations of the gene in 19C4 were mutually exclusive events. In situ hybridization of a P element probe to the chromosomes of v6 demonstrated that P elements reside at a minimum of five sites on the X chromosome. These P element sites correspond to the mutational and breakage hot spots on that chromosome. The combined genetic and cytological data imply that most of the X-linked lethal mutations that occur in M X v6 hybrids are due to local P element action. Consideration of these and other data suggest that v6 is a weak P strain in the P-M system of hybrid dysgenesis and that other Q strains might also be regarded in this way.     

'Exceptional sons' from Drosophila melanogaster mothers carrying a balancer X chromosome

This study reports on exceptional males which are obtained by using Drosophila melanogaster mothers carrying the balancers In(1)FM6 or In(1)FM7 as one of their X chromosomes. The phenomenon was first observed in interspecific crosses between D. melanogaster females and males of its closest relatives which normally produce unisexual female hybrid progeny. Whereas hybrid sons from these crosses die as third instar larvae, the presence of the particular X balancers in the mother allows a low percentage of sons to survive. Similar sterile males are also observed among non-hybrid flies. Data are presented which suggest that the males thus generated could be hyperploid for part of their X chromosome as a result of a meiotic event in their mothers or else they could start life as female zygotes and change sex through a mitotic event at an early stage.     

Reproductive barriers between populations of the cereal rust mite <Emphasis Type="Italic">Abacarus hystrix</Emphasis> confirm their host specialization

Recent studies have shown that certain host populations of the cereal rust mite Abacarus hystrix are highly specialized in their host use and it is likely that reproductive isolation exists between them. Here I verified this expectation by testing for reproductive barriers between ryegrass and quackgrass populations of A. hystrix. I performed reciprocal crosses between individuals from both populations and observed results of crosses. Leaves of the three grass species, ryegrass, quackgrass and wheat, were used as mating arenas. I used two criteria to determine reproductive barriers between strains: the proportion of female progeny and viability of progeny. If studied populations of this haplodiploid species are reproductively isolated a male-biased sex ratio and/or hybrid progeny of reduced viability would be expected. I found that in the presence of quackgrass and ryegrass pre-zygotic barriers between studied populations exists. On wheat I observed asymmetry in reproductive barriers. Between females from quackgrass and males from ryegrass a pre-zygotic barrier existed (only males obtained). However, the opposite reciprocal cross (females from ryegrass and males from quackgrass) produced progeny of both sexes. A male-biased sex ratio and low adult emergence suggests that post-zygotic mechanisms acted here. Low viability of progeny obtained from crosses in which females from ryegrass were engaged suggests that the origin of the female nymph acted as a predictor of hybrid inviability. The pattern of sterility observed in the cereal rust mite indicates that in the presence of three hosts (ryegrass, quackgrass and wheat) pre- and post-zygotic reproductive barriers between quackgrass and ryegrass populations of this mite exist. In addition to host fidelity (which acts as pre-zygotic barrier) there are post-zygotic barriers to gene flow.     

Genetic control of pest Lepidoptera: construction of a balanced lethal strain in Ephestia kuehniella

A genetic method for the suppression of Lepidopteran pests has been investigated in the Mediterranean flour moth, Ephestia kuehniella Zeller. The method is based on the release of males trans-heterozygous for two sex-linked recessive lethal mutations (SLRLMs). In this paper, characteristics of 16 new SLRLMs are presented. The construction of a balanced lethal strain, BL-2, which was the last step to develop the method, is reported. Males of the strain are balanced for two non-allelic SLRLMs, sl-2 and sl-15. Females carry either sl-2 or sl-15 in their Z chromosome and the T(W;Z)2 translocation on their W chromosome. The translocation includes wild-type alleles of both lethal loci so that the females are viable. Matings between males of the BL-2 strain and normal females of the wild-type strain gave 99.74% male progeny. Exceptional females were due to recombination between the sl-2 and sl-15 loci. Thus, males of the BL-2 strain have a potential to suppress wild populations of the pest. Another envisaged use of this method is for an effective sexing technique.     

The relationship between radiation-induced and transposon-induced genetic damage during Drosophila oogenesis

The combined effect of X-irradiation and transposon mobility on the frequencies of X-linked recessive lethals and dominant lethals was investigated in female hybrids in the P-M system of hybrid dysgenesis. X-linked lethals were measured in G2 hybrid dysgenic females whose X chromosome was derived from the M X P cross. To test for additivity or synergism, the mutation rate in irradiated dysgenic females was compared to that of unirradiated females as well as to irradiated nondysgenic hybrid females derived from M X M crosses. Eggs collected for 2 days after irradiation, were represented by the more radiation-sensitive A and B oocytes (about 75%) and the least sensitive C oocytes (about 25%). The production of X-linked lethal events in X-irradiated dysgenic females was 8.1%, as compared to 4.5% in dysgenic controls and 3.4% in irradiated, nondysgenic controls, demonstrating an additive effect of radiation and dysgenesis-induced genetic damage. The effect of irradiation on sterility of dysgenic hybrid females was a negative one, resulting in 20% less sterility than expected from an additive effect. The combined effect of radiation and dysgenesis on dominant lethality tested in A, B and C oocytes of the same hybrid females was synergistic. Egg broods collected for 3.5 days after irradiation showed that significantly more damage was induced in the presence of ionizing radiation in dysgenic females than in their nondysgenic counterparts. This effect was most obvious in B and C oocytes. The synergism observed may be related to the inability of cells to repair the increased number of chromosome breaks induced both by radiation and transposon mobility.     

Transposable Element-Induced Response to Artificial Selection in DROSOPHILA MELANOGASTER

The P family of transposable elements in Drosophila melanogaster transpose with exceptionally high frequency when males from P strains carrying multiple copies of these elements are crossed to females from M strains that lack P elements, but with substantially lower frequency in the reciprocal cross. Transposition is associated with enhanced mutation rates, caused by insertion and deletion of P elements, and chromosome rearrangements. If P element mutagenesis creates additional variation for quantitative traits, accelerated response to artificial selection of progeny of M female female X P male male strain crosses is expected, compared with that from progeny of P female female X M male male strain crosses.--Divergent artificial selection for number of bristles on the last abdominal tergite was carried out for 16 generations among the progeny of P-strain males (Harwich) and M-strain females (Canton-S) and also of M-strain males (Canton-S) and P-strain females (Harwich). Each cross was replicated four times. Average realized heritability of abdominal bristle score for the crosses in which P transposition was expected was 0.244 +/- 0.017, 1.5 times greater than average heritability estimated from crosses in which transposition was expected to be rare (0.163 +/- 0.010). Phenotypic variance of abdominal bristle score increased by a factor of four in lines selected from M female female X P male male crosses when compared with those selected from P female female X M male male hybrids. Not all quantitative genetic variation induced by P elements is additive. A substantial fraction of nonadditive genetic variation is implicated by chromosomal analysis, which demonstrates deleterious fitness effects of the mutations when homozygous.--Several putative "quantitative" mutations were identified from chromosomes extracted from the selected lines; these will form the basis for further investigation at the molecular level of the genes controlling quantitative inheritance.     

Sex-specific effects of hybridization on reproductive fitness in Mytilus

Blue mussels of the genus Mytilus form extensive hybrid zones in the North Atlantic and elsewhere where the distributions of different species overlap. Mytilus species transmit both maternal and paternal mtDNA through egg and sperm, respectively, a process known as doubly uniparental inheritance (DUI), and some females produce offspring with extremely biased sex ratios. These two traits have been shown to be linked and maternally controlled, with sex determination involving nuclear–cytoplasmic interactions. Hybridization has been shown to disrupt DUI mitochondrial inheritance and sex ratio bias; however, the effect of hybridization on reproductive fitness has not previously been examined. We investigated this effect in M. edulis × M. trossulus crosses through histological examination of mature F1 progeny, and spawning of F1 hybrids to monitor survival of their progeny through to the D stage of larval development. For progeny produced from mothers with a strong bias toward female offspring (often 100%) in pure-bred crosses, there was a clear breakdown in female dominance of progeny and significantly more hermaphrodites in the hybrid crosses produced from sperm with the M-tr1 mitotype. We also found significant sex-specific differences among hybrid progeny, with females producing normal eggs while males and hermaphrodites evidenced impaired gonadal development with significantly greater numbers of Sertoli cells, phagocytic hemocytes, and degenerating germ cells, all associated with gonad resorption. Males from crosses where DUI was disrupted and where male progeny were homoplasmic for the female mtDNA were the most severely compromised. Allelic incongruity between maternal and paternal mitotypes in hybrid crosses was associated with significant disruption of male gonadal development.     

Developmental success and reproductive incompatibility among populations of the European red mite,Panonychus ulmi (Acari: Tetranychidae)

Developmental success on 47 species of plant and reproductive compatibility were studied for four strains of the European red mite,Panonychus ulmi (Koch), collected from dwarf bamboo, elm and apple trees. The host range was variable in those strains and there was no plant species on which all four strains were able to reach maturity.Intra-populational crosses in the dwarf-bamboo strain and the two apple strains of mites, as well as crosses between the two apple strains, gave both female and male progeny. Inter-populational crosses between the dwarf-bamboo and apple strains produced only male progeny, as in the case where females of each strain were kept virgin, indicating that these two were reproductively incompatible and had an arrhenotokous reproductive system. When females of the elm strain were crossed with males of the elm, dwarf-bamboo or apple strain, no males were produced, as in the case for virgin females of the elm strain which had a thelytokous reproductive system. It is thus concluded that the strains derived from different host species were reproductively incompatible with one another.     

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.     

Genetics of Factors Affecting the Life History of DROSOPHILA MELANOGASTER. I. Female Productivity

Starting from four basic strains of Drosophila melanogaster, two laboratory strains (cn bw, Tokyo) and two isofemale lines (B-102, B-103) originated from a wild population in Texas, we constructed by repeated backcrosses through females for 20 or more generations a total of 16 strains of all possible combinations between the chromosome sets and cytoplasmic classes. Females from these 16 synthesized strains were then examined for their reproductive performance during their entire life span.---The chromosome set from the cn bw strain was found to associate with the highest female productivity when the age of females was very young, but these females ceased their reproduction and died relatively earlier, resulting in a smaller number of total progeny. The B-102 and B-103 chromosome sets, on the other hand, were associated with the lowest productivity when the females were young, but they lived and continued reproduction longer, resulting in a larger number of total progeny. The Tokyo chromosome set was associated with female productivity intermediate between the other two groups.---Cytoplasmic factors were found to affect the productivity of young females, with the cytoplasm from the cn bw strain associated with the highest productivity. Longevity was not cytoplasmically affected.---There was a clear interaction in female productivity between the Tokyo chromosome set and the cytoplasm from the Texas isofemale lines; the lifetime female productivity, as well as longevity, associated with the Tokyo chromosome set was found to increase considerably when it was substituted into the cytoplasm of the Texas isofemale lines. This chromosome-cytoplasm interaction appeared to be independent of the two systems of hybrid dysgenesis.     

Linkage between sperm abnormality level and major urinary protein phenotype in mice

Segregation of sperm abnormality level and the pattern of major urinary proteins (MUPs) were investigated in F2 and B1 hybrid males obtained from crosses involving two contrasting inbred strains of mice: CBA/Kw (Mup-1a1a, 3.3% abnormal sperm) and C57BL/Kw (Mup-1b1b, 21.9% abnormal sperm). In the progeny of both crosses mean levels of abnormal spermatozoa were significantly higher for males typed as Mup-1b1b than for heterozygous Mup-1a1b males. Moreover, all F2 hybrid males showing very high percentages of abnormal sperm were Mup-1b1b homozygotes. Similarly, among B1 males with a high level of deformed spermatozoa, a statistically significant majority were Mup-1b1b genotypes. Our results suggest that at least two genes which influence sperm abnormality level are segregating in these crosses. Both appear to be recessive for high sperm abnormality level, and one shows weak linkage to Mup-1 on chromosome 4.     

Fitness effects of X chromosome drive in the stalk-eyed fly, Cyrtodiopsis dalmanni

Sex-ratio (SR) males produce predominantly female progeny because most Y chromosome sperm are rendered nonfunctional. The resulting transmission advantage of XSR chromosomes should eventually cause population extinction unless segregation distortion is masked by suppressors or balanced by selection. By screening male stalk-eyed flies, Cyrtodiopsis dalmanni, for brood sex ratio we found unique SR alleles at three X-linked microsatellite loci and used them to determine if SR persists as a balanced polymorphism. We found that XSR/XST females produced more offspring than other genotypes and that SR males had lower sperm precedence and exhibited lower fertility when mating eight females in 24 h. Adult survival was independent of SR genotype but positively correlated with eye span. We infer that the SR polymorphism is likely maintained by a combination of weak overdominance for female fecundity and frequency dependent selection acting on male fertility. Our discovery of two SR haplotypes in the same population in a 10-year period further suggests that this SR polymorphism may be evolving rapidly.     

Characterization of defects in adult germline development and oogenesis of sterile and rescued female hybrids in crosses between Drosophila simulans and Drosophila melanogaster

Crosses between Drosophila melanogaster and D. simulans normally result in progeny that are either inviable or sterile. Recent discovery of strains that rescue these inviability and sterility phenotypes has made it possible to study the developmental basis of reproductive isolation between these two species in greater detail. By producing both rescued and unrescued hybrids and examining the protein product staining patterns of genes known to be involved in early germline development and gametogenesis, we have found that in crosses between D. simulans and D. melanogaster, hybrid female sterility results from the improper control of primordial germline proliferation, germline stem cell maintenance, and cystoblast formation and differentiation during early oogenesis. Rescued hybrid females are fertile, yet they generally have lower amounts of adult germline from the outset and show a premature degeneration of adult germline cells with age. In addition, older rescued hybrid females also exhibit mutant egg phenotypes associated with defects in dorso-ventral patterning which may result from the improper partitioning of cytoplasmic factors during early oogenesis that could stem from the early defect. Although a variety of germline and oogenic defects are described for the hybrid females, all of them can potentially result from the same underlying primary defect. Hybrid males from these same crosses, on the other hand, have no detectable germline in adult reproductive tissues, even when hybrid sterility rescue strains are used, indicating that male sterility and female sterility stem from distinctly different developmental defects.     

Sex and death in birds: a model of dosage compensation that predicts lethality of sex chromosome aneuploids

Birds show female heterogamety, with ZZ males and ZW females. It is still not clear whether the W is female-determining, or whether two doses of the Z chromosomes are male-determining, or both. This question could easily be settled by the sexual phenotypes of ZZW and ZO birds, in the same way that the sexual phenotypes of XXY and XO showed that the Y is male determining in humans, but that the dosage of an X-borne gene determines sex in Drosophila. However, despite extensive searches, no ZZW or ZO diploid birds have been satisfactorily documented, so we must assume that these genotypes are embryonic lethals. Given that ZW and ZZ are viable and the W contains few genes it is not clear why this should be so. Here I propose that sex chromosome aneuploids are lethal in chicken because, to achieve dosage compensation, a locus on the W chromosome controls the upregulation of genes on the Z in ZW females. ZO birds would therefore have only half the normal dose of Z-linked gene product and ZZW would have twice the amount, both of which would undoubtedly be incompatible with life. Reports of other aneuploids and triploids are also consistent with this hypothesis.     

High Mutability in Male Hybrids of DROSOPHILA MELANOGASTER

The frequencies of sex-linked lethal mutations arising in hybrid male offspring from various crosses and in nonhybrid controls were determined. The hybrids were produced by crossing representative strains of the P-M system of hybrid dysgenesis in all possible combinations. Males from the cross of P males x M females had a mutation rate about 15 times higher than that of nonhybrid males from the P strain. Genetically identical males from the reciprocal cross had a mutation rate 3 to 4 times that of the nonhybrids. For crosses involving a Q strain, a significant increase in the mutation rate was detected in males produced by matings of Q males with M females. No increase was observed in genetically identical males from the reciprocal mating. Crosses between P and Q strains gave male hybrids with mutation rates not different from those of nonhybrids. Many of the lethals that occurred in hybrids from the cross of P males x M females appeared to be unstable; fewer lethals that arose in hybrids from the cross of Q males x M females were unstable. The relationship between P and Q strains is discussed with respect to a model of mutation induction in dysgenic hybrids.     

Postzygotic isolation between the two European subspecies of the house mouse: estimates from fertility patterns in wild and laboratory-bred hybrids

We assessed the fertility (reproductive success, litter size, testis weight, spermatocyte-to-spermatid ratio) of F₁s and backcrosses between different wild-derived outbred and inbred strains of two mouse subspecies, Mus musculus domesticus and M. m. musculus . A significant proportion of the F₁ females between the outbred crosses did not reproduce, suggesting that female infertility was present. As the spermatocyte-to-spermatid ratio was correlated with testis weight, the latter was used to attribute a sterile vs. fertile phenotype to all males. Segregation proportions in the backcrosses of F₁ females yielded 11 (inbred) to 17% (outbred) sterile males, suggesting the contribution of two to three major genetic factors to hybrid male sterility. Only one direction of cross between the inbred strains produced sterile F₁ males, indicating that one factor was borne by the musculus X-chromosome. No such differences were observed between reciprocal crosses in the outbred strains. The involvement of the X chromosome in male sterility thus could not be assessed, but its contribution appears likely given the limited introgression of X-linked markers through the hybrid zone between the subspecies. However, we observed no sterile phenotypes in wild males from the hybrid zone, although testis weight tended to decrease in the centre of the transect.  © 2005 The Linnean Society of London, Biological Journal of the Linnean Society , 2005, 84 , 379–393.     

Determination of the probable ontogenic time of action of lethal factors on thesecond chromosome of Drosophila melanogaster derived from a natural population

From the study of the genetic load of second chromosome factors in a natural population of Drosophila melanogaster, 15 lethal-bearing strains were recovered and maintained in the laboratory balanced against Ins (2L+2R), Cy, L⁴. For each lethal factor, the probable time of action during development was determined by the appearance of a sharp reduction, at any given stage, in the frequency of individuals compared to that expected in the absence of the lethal factor. Carried out in this way, the analysis suggested that sevne were embryonic lethals, two larval lethals and three pupal lethals. Additionally, three gave no evidence of affecting any of the above-mentioned stages; these are interpreted as gametic lethals.     

Scope for genetic enhancement of the parasitisation potential of four native strains of Trichogrammatoidea sp. nr. lutea Girault (Hymenoptera: Trichogrammatidae) in Kenya

In response to emerging interest in commercial mass production of Trichogramma for Helicoverpa armigera biocontrol in eastern Africa, laboratory experiments were undertaken to assess the scope for genetic enhancement of the parasitisation potential of native strains of the local common trichogrammatid species, Trichogrammatoidea sp. nr. lutea. Four promising strains (ex-Kilifi – Kilifi District, ex-Kwa Chai – Kibwezi District, ex-Rarieda – Bondo District and ex-Ebuhayi, Kakamega District) were tested for cross-mating in reciprocal combinations with focus on fecundity and progeny female ratio. While all the crosses resulted in F1 progeny of both sexes, significant differences were observed between homogamic and reciprocal heterogamic crosses in fecundity, progeny production, proportion of female progeny and adult longevity. Among all the crosses, the cross between ex-Rarieda strain females and ex-Kilifi strain males resulted in progeny that was significantly superior in fecundity and progeny female ratio. Conversely, Kilifi strain females crossed to males from ex-Rarieda strain gave rise to progeny with relatively low fecundity and female ratio. There were significant differences between homogamic crosses and most reciprocal heterogamic crosses in the major biological attributes. Genotypic and phenotypic variance-covariance matrices generated for six life-history traits showed high positive correlations for most traits in both inbred (P<0.05) and reciprocal heterogamic crosses (P<0.05 and P<0.001). Fecundity and number of female offspring were the most important factors in the heterogamic crosses. The results confirmed the scope for genetic enhancement through inter-strain crossing for improving the field impact potential of T. sp. nr. lutea being targeted for commercial mass production.