MITOCHONDRIAL-DNA ANALYSES AND THE ORIGIN AND RELATIVE AGE OF PARTHENOGENETIC LIZARDS (GENUS CNEMIDOPHORUS). III. C. VELOX AND C. EXSANGUIS

Mitochondrial DNAs (mtDNAs) of two unisexual, parthenogenetically reproducing species of whiptail lizards (Cnemidophorus velox and C. exsanguis) and their bisexual relatives were compared by restriction-enzyme analysis to assess levels of mtDNA variation and to establish the maternal ancestry of the unisexuals. No cleavage-site differences were found to be diagnostic between C. velox and C. exsanguis mtDNAs, suggesting an ancestry rooted in the same maternal lineage. The mtDNA of the unisexuals is relatively homogeneous, indicating that these lineages are of recent origin. Phylogenetic analysis revealed that the maternal ancestor of both C. velox and C. exsanguis was most probably C. burti stictogrammus, C. costatus barrancorum, or an unidentified taxon closely related to them. In addition, the mtDNA analyses demonstrate conclusively that the triploid species C. velox could not have been formed by the fertilization of an unreduced (diploid) C. inornatus egg, further strengthening the hypothesis that parthenogenesis in Cnemidophorus results from hybridization.     

Additional notes on requirements of captive whiptail lizards (Cnemidophorus), with emphasis on ultraviolet radiation

Since 1972 we have maintained a colony of parthenogenetic Cnemidophorus exsanguis, which has reached the seventh generation in our laboratory. This paper reports improved husbandry techniques that allow maintenance of healthy, reproducing colonies of this and additional species of Cnemidophorus. In addition, aspects of the physiology of calcium regulation and metabolic bone disease are discussed in relation to the captive rearing of lizards through many generations.     

Variation in the sexual behaviour of Achnanthes longipes (Bacillariophyta). III. Progeny of crosses between monoecious and unisexual clones

The allogamous raphid diatom Achnanthes longipes C. A. Agardh possesses a complex breeding system involving interactions between three types of clone: monoecious, unisexual and bisexual. Previous studies showed that these three types can be crossed with each other, with a tendency for sexual characteristics to be inherited: inbred monoecious lineages gave rise to monoecious or, very rarely, to bisexual clones, while inbred unisexual lineages yielded unisexual and bisexual clones. The current paper reports on the progeny of crosses between monoecious and unisexual clones and their inbred offspring. All three types of clone appeared in the F₁ and F₂, although unisexual clones of opposite sex to the parental clone were not found. Inbreeding depression was observed and also a tendency for ‘normal’ auxosporulation (producing two auxospores per pair of gametangia) to be replaced by ‘reduced’ or ‘intermediate’ auxosporulation (producing one auxospore per pair). In addition, patterns of incompatibility were observed that were not seen during earlier studies of clones isolated directly from nature. These included the inability of some F₁ clones to mate with each other, in spite of compatibility with all other clones examined (unisexual, bisexual and monoecious).     

THE EVOLUTIONARY HISTORY OF PARTHENOGENETIC CNEMIDOPHORUS LEMNISCATUS (SAURIA: TEIIDAE). II. MATERNAL ORIGIN AND AGE INFERRED FROM MITOCHONDRIAL DNA ANALYSES

Restriction endonuclease analyses were performed on mitochondrial DNAs (mtDNAs) representing unisexual parthenogenetic (cytotypes A, B, and C) and bisexual (cytotypes D and E) populations of Amazonian lizards presently regarded as Cnemidophorus lemniscatus. The results of mtDNA cleavage map comparisons among these C. lemniscatus indicated that (1) there was no cleavage site variation among the unisexuals, (2) mtDNAs from the bisexual cytotypes D and E differed in sequence from one another by about 13%, and (3) mtDNAs from cytotypes A–C differed from those of cytotype D by about 5% and from those of cytotype E by about 13%. Higher resolution restriction fragment size comparisons confirmed the high degree of similarity among the unisexual mtDNAs, but identified 12 cleavage site variants among the 13 cytotype D mtDNAs examined. Both cladistic and phenetic (UPGMA) analyses of the data indicate that the unisexual and cytotype D mtDNAs form a single clade, suggesting that a female of cytotype D was the maternal progenitor of the unisexuals. The similarity among the unisexual mtDNAs and the variability among those of cytotype D suggest that the three unisexual cytotypes arose recently from a common maternal lineage. The mtDNA variability observed among cytotype D individuals has a strong geographic component, suggesting that the unisexuals arose from one or a few geographically proximal populations. The mtDNA comparisons also support the conclusion, based on allozyme comparisons (Sites et al., 1990, this issue), that cytotypes D and E, although presently allocated to C. lemniscatus, are separate species.     

THE EVOLUTIONARY HISTORY OF PARTHENOGENETIC CNEMIDOPHORUS LEMNISCATUS (SAURIA,TEIIDAE). I. EVIDENCE FOR A HYBRID ORIGIN

Chromosomes and allozymes were studied from chromosomally distinct unisexual (races B and C) and bisexual (races D and E) populations of the teiid lizard Cnemidophorus lemniscatus, and from selected outgroup taxa (C. murinus, C. nigricolor, Ameiva ameiva, and A. auberi). Karyotyping confirmed the racial identity of individuals and showed that the chromosomal composition of populations at specific localities has remained the same for 20 years. All individuals of both unisexual populations were heterozygous for a pericentric inversion that distinguishes D and E bisexuals. Also, the unisexuals were all heterozygous for 8 of 11 protein loci for which D and E were fixed or nearly fixed for different alleles. Most of these alleles represent derived states relative to the other Cnemidophorus and Ameiva analyzed, and the fixed heterozygote condition at these nine markers provides unequivocal support for the hypothesis that the unisexual C. lemniscatus arose by hybridization between ancestors genetically similar to extant D and E populations. At the remaining three loci for which D and E show fixed differences, the unisexuals were homozygous rather than heterozygous. This suggests that either (1) allozymes have been lost by mutations to null, silent, or convergent mobility states, (2) ancestral genotypes were similar to but not identical with the extant D and E races, and/or (3) limited recombination may occur between unisexual genomes. Allozyme-based genetic distances between D and E were large, suggesting that bisexual races D and E are genetically isolated; each race should be accorded full species status. This conclusion is supported by the absence of any clear biochemical evidence for their monophyly with respect to the other Cnemidophorus examined. Cladistic analyses of 17 phylogenetically informative loci revealed two equally parsimonious shortest trees, one supporting monophyly and the other paraphyly of the C. lemniscatus complex. Further testing of the monophyly of C. lemniscatus requires additional data. With the present study, the evidence that all parthenogenetic Cnemidophorus are of hybrid origin is complete.     

Sex determination and evolution of unisexuality in the Conchostraca

Field collected or laboratory-reared samples of 60 species of conchostracans (representing all extant genera) indicate that males and females are equally common in most species. Deviations from this pattern occur in four lineages.Cyzicus andLeptestheria each include at least one unisexual species; many species of Limnadiinae are either unisexual or characterized by female-biased sex ratios; and Cyclestheriidae are either unisexual or express males in the later generations of their life cycles. Laboratory studies indicate that species with sex ratios near unity are gonochoric (obligately sexual), whereas females in species with female-biased sex ratios are capable of both outcrossing and selfing modes of reproduction. Phylogenetic analysis of patterns of reproduction suggest that sexual reproduction is the primitive condition. Genetic analysis of sexual species indicate that gender is determined by one or a few genetic factors and that the male-determining allele is recessive. The inheritance of gender in androdioecious species (where females are capable of self-fertilization) is similar to that in sexual species. Androdioecy is likely to be the intermediate stage between obligately sexual reproduction and unisexuality in the Limnadiinae. The phylogenetic distribution of sex ratio variation suggests that unisexuality in Cyzicidae, Leptestheriidae, and Cyclestheriidae has arisen independently of that in the Limnadiinae and that these cases have evolved by different evolutionary pathways.     

Graptodytes exsanguis (Bedel, 1925) n. stat., a newly recognised species of diving beetle from North Africa,Corsica and Sardinia,with notes on other taxa of the varius/ignotus complex (Coleoptera: Dytiscidae)

Summary

A species of the genus Graptodytes Seidlitz, 1887 (varius/ignotus complex) is newly recognised and described based on recently sampled specimens from Corsica and Sardinia, and material in various collections from Algeria, Tunisia, Corsica and Sardinia. The name exsanguis, created by Bedel in 1925 for a variety of what he then called Hydroporus varius Aubé, 1838, is shown to be available for the newly recognised species, therefore named Graptodytes exsanguis (Bedel, 1925) n. stat. (type locality: Algeria, La Calle – nowadays El Kala). This species is similar in size and colour pattern to G. fractus (Sharp, 1882), from which it can be distinguished by strong differentiation of the male protarsal claws, features of the habitus and elytral punctation, and shape of the median lobe of aedeagus. Morphological features of G. exsanguis and G. fractus are illustrated, and pictures showing for each species the variability of habitus and median lobe are provided. For comparison, the habitus and median lobes of other species of the varius/ignotus complex are also illustrated. Habitat preferences of G. exsanguis are described based on field observations in Corsica and Sardinia, and a distribution map of validated records is presented. Analysis of a partial CO1 alignment yielded a basal and divergent position within the varius/ignotus complex for the sequenced G. exsanguis haplotypes from Corsica and Sardinia. Lectotypes are designated for Hydroporus varius var. exsanguis Bedel, 1925, Hydroporus fractus Sharp, 1882 and Graptodytes ignotus var. fallaciosus Guignot, 1932, and the latter is established as a junior synonym of G. fractus.     

Effects of winter temperatures on the sex ratios ofAphytis melinus [Hym.: Aphelinidae] in the San Joaquin Valley of California

Laboratory bioassays of overwintering field populations of the California red scale parasitoidAphytis melinus DeBach showed a shift in sex ratio of F₁ progeny after prolonged exposure to winter temperatures in Tulare County, California. In 7 of 8 tests, short-term winter field exposure (<75 days) did not result in a significant male bias in sex ratios of F₁ progeny when compared to sex ratios observed in laboratory cultures maintained at 27°C. Long term exposure (109–139 days), however, resulted in significantly (P<0.05) higher male bias in F₁ sex ratios in 5 of 6 tests. These results from field populations ofA. melinus agree with conclusions of low temperature laboratory tests onAphytis species (lingnanensis, melinus) by other workers. Mention of a proprietary or commercial product in this paper does not constitute an endorsement of this product by the University of California.     

L-chromosome inheritance and the problem of chromosome “imprinting” in Sciara (Sciaridae,Diptera)

The loss of the L-chromosomes from the germ line of a newly-collected strain of Sciara impatiens was used as a genetic marker to test whether the supernumeraries in this genus obey the rule of origin which governs the behavior of the regular homologues at first meiotic anaphase in the male. Reciprocal crosses between a normal strain which possessed L's and the no-L line yielded F₁ males whose germ cells contained L-chromosomes of paternal or of maternal origin. Cytological study of the gonads of these males showed that in all instances, irrespective of their origin, the L-chromosomes were conserved. To demonstrate that the L's thus conserved are not lost subsequently during spermiogenesis, the F₁ males derived from normal fathers were bred to females from the no-L line; the gonads of the test-cross progeny showed that these individuals had inherited L-chromosomes. The failure of the paternal L's to obey the rule of origin during spermatogenesis indicates that the supernumeraries in this genus are not subject to the reversible imprinting which the regular complement undergoes. And the genetic effects of exclusively paternal or maternal L-chromosomes on germ cell development and maturation are indistinguishable. Concomitant with the loss of L's from the germ line, the newly-collected strain of S. impatiens underwent an abrupt alteration from unisexual families to variable sex ratios. These ratios are discussed in relation to the problem of sex determination in S. ocellaris, a species which sustained L-chromosome loss during the course of evolution. Discussed also are some new unpublished data which reveal, unexpectedly, a genetic affinity between ocellaris and impatiens. The no-L line of impatiens, which has been kept in laboratory culture for approximately four years, provides a unique opportunity to examine the conditions under which the supernumeraries are retained or lost and also to determine more precisely the nature of their genetic effects.It is a pleasure to acknowledge support by the National Science FoundationIt is a pleasure to acknowledge support by the National Science Foundation     

Status of the reproductive system of bony fish from the Teterev River and the Kiev Reservoir 20 years after the Chernobyl accident

The status of the reproductive system is investigated in five species of Teleostei from the Teterev River and the Kiev Reservoir (Ukraine), descendants of F₄2–F₇ generations of fish exposed to radiation from the accident at the Chernobyl nuclear power plant. Twenty years after the accident, a wide range of morphofuinctional anomalies of this system is present in fish. The most significant of them are: asymmetry and anomalous morphology of gonads, proliferation of connective tissue—sterilization, mass destruction of follicular and sex cells of various developmental stages, and hermaphroditism. Among the investigated species, disturbances of reproductive glands were preset to the highest extent in pike Esox lucius and crucian carp Carassius carassius. In the series of investigated generations the maximum frequency of occurrence of gonad anomalies was noted in F₄ and F₅, due to prolonged mutagenesis (Dubinin, 1986).     

Geographic patterns of postzygotic isolation between two closely related widespread dung fly species (Sepsis cynipsea and Sepsis neocynipsea; Diptera: Sepsidae)

Identifying the contribution of pre‐ and postzygotic barriers to gene flow is a key goal of speciation research. The widespread dung fly species Sepsis cynipsea and Sepsis neocynipsea offer great potential for studying the speciation process over a range of opportunities for gene exchange within and across sister species (cross‐continental allopatry, continental parapatry and sympatry). We examined the role of postcopulatory isolating barriers by comparing female fecundity and egg‐to‐adult viability of F₁ and F₂ hybrids, as well as backcrosses of F₁ hybrids with the parental species, via replicated crosses of sym‐, para‐ and allopatric populations. Egg‐to‐adult viability was strongly but not totally suppressed in hybrids, and offspring production approached nil in the F₂ generation (hybrid breakdown), indicating yet unspecified intrinsic incompatibilities. Viable F₁ hybrid offspring showed almost absolute male (the heterogametic sex) sterility while females remained largely fertile, in accordance with Haldane's rule. Hybridization between the two species in European areas of sympatry (Swiss Alps) indicated only minor reinforcement based on fecundity traits. Crossing geographically isolated European and North American S. neocynipsea showed similar albeit weaker isolating barriers that are most easily explained by random genetic drift. We conclude that in this system with a biogeographic continuum of reproductive barriers, speciation is mediated primarily by genetic drift following dispersal of flies over a wide (allopatric) geographic range, with some role of natural or sexual selection in incidental or direct reinforcement of incompatibility mechanisms in areas of European sympatry. S(ubs)pecies status of continental S. neocynipsea appears warranted.     

Extinction of marine renewable resources: a demographic analysis

A simple (deterministic) population dynamics model is used to examine the level of fishing mortality at which a population is expected to be driven to extinction (F _crash). Values for F _crash are determined for six marine fish species (Cape hake, blue grenadier, blue warehou, school shark, gummy shark, and orange roughy) subject to commercial harvest. The sensitivity of the value of F _crash to changing the selectivity pattern of the fishery and the relationship between the size of the reproductive component of the population and subsequent births to allow for depensatory effects is examined. F _crash is greatest for highly productive species and when the fishery does not target immature animals. The ratio of F _crash to the fishing mortality at which maximum sustainable yield is achieved, F _MSY, is a decreasing function of the productivity of the population. The possibility of depensation has little impact on F _MSY but can substantially reduce the ratio F _crash/F _MSY. A series of stochastic simulations is conducted to assess the probability of detecting that the extent of fishing exceeds F _crash using the current IUCN A criterion. The results of these simulations are also used to determine the probability that the IUCN A criteria will be triggered when fishing takes place at F _MSY. The results indicate that there are substantial probabilities of incorrectly identifying species being harvested at F _MSY as being threatened during the 'fishing down' phase and also of not identifying species actually at risk of extinction if fishing mortality is not reduced. Received: January 14, 1999 / Accepted: April 27, 1999     

Comparisons of body sizes at sexual maturity and at sex change in the parrotfishes of Hawaii: input needed for management regulations and stock assessments

First estimates of sex allocation patterns and body size‐at‐sexual maturity and at protogynous sex change are presented for the five major (including one endemic) species of parrotfishes of Hawaii. Median body size at initial maturation as a female (L_M50) and at protogynous sex change from adult female to adult male (L_Δ50) varied greatly among the five species. Estimates of L_M50 were about 14, 17, 24, 34 and 35 cm fork length (L_F) in palenose Scarus psittacus, Pacific bullethead Chlorurus spilurus, stareye Calotomus carolinus, spectacled Chlorurus perspicillatus and redlip parrotfish Scarus rubroviolaceus. Values of L_Δ50 were c. 23, 27, 37, 46 and 47 cm L_F in the respective species. Length at female maturation was proportional to maximum body size (L_max) of the respective species, ranging from 50 to 72% and averaging 62% of L_max across species. L_Δ50 was also proportional to L_max, ranging from 82 to 97% and averaging 92%. Males of both pairs of Scarus and Chlorurus spp. reported here are diandric. Only one of the five major species (C. carolinus) is functionally monandric, with either all or nearly all males secondarily derived from adult females. The broadly differing absolute body sizes at sexual maturation and at sex change among the five species have important implications for improving regulatory size limits for parrotfishes in the State of Hawaii, where parrotfish species have historically been managed based on a single minimum size limit of 30·5 cm L_F. This study provides a model demonstration of why catch data for parrotfishes, and other size‐structured reef‐fish populations, should be recorded either by species or by functional size‐groups of species that allow setting more meaningful minimum size limits.     

Expression of AODEF,a B-functional MADS-box gene,in stamens and inner tepals of the dioecious species Asparagus officinalis L.

Garden asparagus (Asparagus officinalis L.) is a dioecious species with male and female flowers on separate unisexual individuals. Since B- and C-functional MADS-box genes specify male and female reproductive organs, it is important to characterize these genes to clarify the mechanism of sex determination in monoecious and dioecious species. In this study, we isolated and characterized AODEF gene, a B-functional gene in the development of male and female flowers of A. officinalis. Southern hybridization identified a single copy of AODEF gene in asparagus genome. Northern blot analysis showed that this gene was specifically expressed in flower buds and not in vegetative tissues. In situ hybridization showed that during early hermaphrodite stages, AODEFgene was expressed in the inner tepal and stamen whorls (whorls 2 and 3, respectively), but not in the outer tepals (whorl 1), in both male and female flowers. In late unisexual developmental stages, the expression of AODEF gene was still detected in the inner tepals and stamens of male flowers, but the expression was reduced in whorls 2 and 3 of female flowers. Our results suggest that AODEF gene is probably not involved in tepal development in asparagus and that the expression of AODEF gene is probably controlled directly or indirectly by sex determination gene in the late developmental stages.     

Behavioral responses of Ceratitis capitata flies to bait spray droplets and natural food

We usedF-statistics to quantify the population structure of two sympatric species of leaf beetles,Oreina cacaliae andO. speciosissima (Chrysomelinae, Coleoptera), which share the same microhabitat since they feed on the same herbaceous host plants. We measured genetic differentiation at six allozyme loci 1) among populations separated by relatively small distances (40 to 250 kilometers), 2) within each population, and 3) between sexes within populations. For both species, the populations studied are not panmictic. For each population, heterozygosities are relatively high, but the observed heterozygosities are generally lower than the expected values. Overall, within-population differentiation is high and similar for both species (F _is=0.326 forO. cacaliae and 0.332 forO. speciosissima). Additionally, populations of both species are highly differentiated (F _st=0.234 versus 0.051 forO. speciosissima). ForO. cacaliae,F _is andF _st are greater among females than among males, while forO. speciosissima,F _st is sustantially greater among the males whileF _is is slightly greater among males. Differences in gene frequency among the sexes were statistically tested using a modifiedF _st with sex as the defining category, and the sexes differed significantly with the exception of one population inO. cacaliae. Possible explanations for this difference are discussed.     

Home ranges of parthenogenetic and bisexual species in a community of Darevskia lizards (Reptilia: Lacertidae)

We analysed the home ranges of a community of Darevskia rock lizards composed of a bisexual species (D. valentini), two parthenogens (D. armeniaca and D. unisexualis), and two backcross forms between bisexual and unisexual forms. We estimated home range areas of ink-marked, GPS-located lizards using Minimum Convex Polygon (MCP) and 95% of the locations for those individuals with five or more sightings. The bisexual D. valentini was the species with the largest home ranges, distances travelled, and the most intersections. No differences between unisexual species and backcrosses were recorded for any comparison. In males, home range size and perimeter were related to morphological characteristics. Contrary to what has been described in allopatry, unisexual species showed smaller home ranges and fewer overlaps than sympatric bisexual species. We tentatively suggest that the presence of potential bisexual partners might increase sexual competition among parthenogenetic females while differences in habitat use should also be considered.     

MITOCHONDRIAL-DNA DIVERSITY AND THE ORIGIN OF THE MENIDIA CLARKHUBBSI COMPLEX OF UNISEXUAL FISHES (ATHERINIDAE)

Restriction-endonuclease analysis of mitochondrial DNA (mtDNA) of the unisexual M. clarkhubbsi complex and close sexual relatives indicated that the unisexual complex arose through multiple, nonreciprocal hybridizations involving females of M. peninsulae. High-resolution analyses using restriction endonucleases that cleave at 4-bp sites revealed mtDNA sequence diversity that was low among unisexuals but high among individuals of M. peninsulae. The identification of M. peninsulae as a parent of the unisexuals conflicts with some details of previous allozyme comparisons. One possibility is that the unisexuals were derived from hybridization involving M. beryllina and a recently extinct form of M. peninsulae. In contrast to the unisexuals, contemporary hybrids of M. peninsulae and M. beryllina are formed by reciprocal matings. The origins of extant unisexual lineages from nonreciprocal hybridizations, together with their low mtDNA diversity relative to the maternal ancestor, implies strong constraints on origins of unisexuality via hybridization. Data on reproduction by contemporary F₁ hybrids reveal one form of genetic/developmental constraint: M. peninsulae and M. beryllina may now have diverged beyond the point where the hybrid origin of new unisexual lineages is possible.     

GENETIC EVIDENCE FOR MULTIPLE ORIGINS OF DIOECY IN THE HAWAIIAN SHRUB WIKSTROEMIA (THYMELAEACEAE)

Dioecy is unusually common in the Hawaiian Islands, yet little is known about the evolutionary biology of this breeding system. A native shrub, Wikstroemia, has an unusually diverse array of breeding systems: two forms of dioecy, cryptic and morphological dioecy, as well as hermaphroditism (perfect flowers). The existence of two forms of dioecy is significant for three reasons: 1) the presence of cryptic unisexuals that are functionally unisexual, but retain the appearance of hermaphroditism in both sexes, is strong evidence for the ancestral status of hermaphroditism; 2) the production of nonfunctional pollen, by female cryptic unisexuals, is a new instance of a phenomenon which has previously been reported for a few other species; 3) the two forms of dioecy are morphological markers which are useful in hybridization studies for tracing the genetic basis of their inheritance. Crosses were made between cryptically unisexual individuals (C), between morphologically unisexual individuals (M), and between the two types of unisexuality. The offspring of crosses between individuals with the same sex type usually resulted in offspring with that sex type, but most of the progeny of between-sex type crosses were, unexpectedly, perfect-flowered hermaphrodites. These results show that genetic control of sex determination is not homologous in all populations, suggesting that dioecy has evolved at least twice in Hawaiian Wikstroemia. The genetic data further suggest that males are the heterozygous sex.     

Behavioral Components of Sex Role Reversal in the Tettigoniid Bushcricket Ephippiger ephippiger

Facultative sex role reversal is found in species of tettigoniid bushcrickets in which males invest heavily in matings or offspring by producing large, nutritious spermatophores. On high-quality diets such species show conventional sex roles, but under low-quality diets males become the choosier sex. Comparative work suggests that Ephippiger ephippiger (Tettigoniidae, Ephippigerinae) has one of the largest described spermatophores (up to 40% of the male body weight). Here we examine the behavior of this species under variable diet conditions in the laboratory and find evidence of sex role reversal under poor-quality diet conditions. We also examine the behavioral components of sex role reversal. In the reversed condition, rejections of mating attempts are almost solely by the male and contests are almost solely between females. Role-reversed males sing less frequently and at a much reduced intensity. We use geographic variation in the calling song of this species to assess the strength of female phonotactic discrimination between races. This is not significantly reduced despite sex role reversal. We therefore suggest that the male and female aspects of the acoustic sexual communication system differ in that male components change most during facultative sex role reversal.