Chromosomal rearrangements directly cause underdominant F1 pollen sterility in Mimulus lewisii–Mimulus cardinalis hybrids

Chromosomal rearrangements can contribute to the evolution of postzygotic reproductive isolation directly, by disrupting meiosis in F₁ hybrids, or indirectly, by suppressing recombination among genic incompatibilities. Because direct effects of rearrangements on fertility imply fitness costs during their spread, understanding the mechanism of F₁ hybrid sterility is integral to reconstructing the role(s) of rearrangements in speciation. In hybrids between monkeyflowers Mimulus cardinalis and Mimulus lewisii, rearrangements contain all quantitative trait loci (QTLs) for both premating barriers and pollen sterility, suggesting that they may have facilitated speciation in this model system. We used artificial chromosome doubling and comparative mapping to test whether heterozygous rearrangements directly cause underdominant male sterility in M. lewisii–M. cardinalis hybrids. Consistent with a direct chromosomal basis for hybrid sterility, synthetic tetraploid F₁s showed highly restored fertility (83.4% pollen fertility) relative to diploids F₁s (36.0%). Additional mapping with Mimulus parishii–M. cardinalis and M. parishii–M. lewisii hybrids demonstrated that underdominant male sterility is caused by one M. lewisii specific and one M. cardinalis specific reciprocal translocation, but that inversions had no direct effects on fertility. We discuss the importance of translocations as causes of reproductive isolation, and consider models for how underdominant rearrangements spread and fix despite intrinsic fitness costs.     

RELATIONSHIP OF WOODY PARTHENIUM ARGENTATUM AND HERBACEOUS P. HISPIDUM VAR. AURICULATUM (ASTERACEAE)

Interspecific hybrids between woody Parthenium argentatum Gray (guayule), native to Mexico and Southwest Texas, and herbaceous perennial P. hispidum var. auriculatum (Britton) Rollins, native to the United States, were obtained successfully. The F₁ hybrids were intermediate for most morphological characters with the exception of the short woody stem, yellow pollen color, and the trichome morphology. Chromosome counts revealed the presence of 2n = 36 A-chromosomes in P. argentatum. The same number of A-chromosomes and four B-chromosomes were found in P. hispidum var. auriculatum. Observations of pollen mother cells showed regular meiosis in both parental species. At diakinesis, chiasmata averaged 1.12 and 1.24 per bivalent for P. argentatum and P. hispidum var. auriculatum, respectively. Meiotic behavior of the F₁ hybrids was irregular. F₁ hybrids averaged 4.43 univalents at metaphase I, 1.95 laggards at anaphase I, and 1.62 micronuclei at the tetrad stage. The low pollen stainability (5.1%) in the F₁ hybrids and the limited number of viable BC₁ seeds (4.07%) may be reflections of the irregular meiosis. Although these primary hybrids are partially fertile, they can be used to introduce desirable characteristics of P. hispidum var. auriculatum, such as herbaceous perennial habit, regrowth ability, and cold tolerance into guayule.     

THE TAXONOMIC IMPLICATIONS OF POLLEN MORPHOLOGY IN SOME SOUTH AMERICAN SPECIES OF MIMULUS (SCROPHULARIACEAE)

The pollen grains of South American hexaploid representatives of the Mimulus glabratus complex, section Simiolus, and M. bridgesii, a South American species sometimes included in this complex and sometimes in section Paradanthus, were examined by light and scanning electron microscopy. Mimulus bridgesii has (6-) 4–5-zoniaperturate grains nearly identical to those observed in a complex of species in section Paradanthus. South American members of the M. glabratus complex (M. bridgesii excluded) have the irregularly synaperturate, ± spiraperturate pollen typical of section Simiolus (sensu Grant). Treatment of M. bridgesii as a palynologically aberrant member of section Simiolus is not supported by what is known about the evolution of pollen types in Mimulus and the evolution and dispersal of the M. glabratus complex.     

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.     

INTERSPECIFIC CROSSES IN KALMIA

Interspecific crosses among Kalmia species are reported for the first time. All possible crosses between K. angustifolia, K. cuneata, K. latifolia, K. hirsuta, K. polifolia var. rosmarinifolia and K. p. var. microphylla were attempted. Several crosses yielded no hybrids, many produced albino or yellow-green inviable seedlings, and six combinations produced at least a few seedlings with near-normal growth habit. Stylar inhibition of pollen tube growth prevented some species from hybridizing; reciprocal differences in crossability and pollen tube growth were noted. Kalmia species are highly heterozygous, but no evidence was found within a species for geographic variation of combining ability except for the two varieties of K. polifolia which had different crossing patterns with the other species. The anthocyaninless form of K. angustifolia, in contrast to the normal, has unique combining ability with K. latifolia. No genetic evidence was found to support the placement of K. hirsuta in another genus (Kalmiella) or to designate K. angustifolia var. caroliniana as a distinct species. Use of all the species to breed improved ornamentals is limited by genetic barriers and sterility of F₁ hybrids.     

Airborne‐pollen pool and mating pattern in a hybrid zone between Pinus pumila and P. parviflora var. pentaphylla

The reproductive isolation barriers and the mating patterns among Pinus pumila, P. parviflora var. pentaphylla and their hybrids were examined by flowering phenology and genetic assays of three life stages: airborne‐pollen grains, adults and seeds, in a hybrid zone on Mount Apoi, Hokkaido, Japan. Chloroplast DNA composition of the airborne‐pollen was determined by single‐pollen polymerase chain reaction. Mating patterns were analysed by estimating the molecular hybrid index of the seed parent, their seed embryos and pollen parents. The observation of flowering phenology showed that the flowering of P. pumila precedes that of P. parviflora var. pentaphylla by about 6 to 10 days within the same altitudinal ranges. Although this prezygotic isolation barrier is effective, the genetic assay of airborne‐pollen showed that the two pine species, particularly P. pumila, still have chances to form F₁ hybrid seeds. Both parental species showed a strong assortative mating pattern; F₁ seeds were found in only 1.4% of seeds from P. pumila mother trees and not at all in P. parviflora var. pentaphylla. The assortative mating was concluded as the combined result of flowering time differentiation and cross‐incompatibility. In contrast to the parental species, hybrids were fertilized evenly by the two parental species and themselves. The breakdown of prezygotic barriers (intermediate flowering phenology) and cross‐incompatibility may account for the unselective mating. It is suggested that introgression is ongoing on Mount Apoi through backcrossing between hybrids and parental species, despite strong isolation barriers between the parental species.     

EARLY STAGES IN THE FORMATION OF INTERNAL BARRIERS TO GENE EXCHANGE BETWEEN DIPLOID SPECIES OF SOLANUM

Grun , Paul . (Pennsylvania State U., University Park.) Early stages in the formation of internal barriers to gene exchange between diploid species of Solanum. Amer. Jour. Bot. 48(1): 79–89. Illus. 1961.—Internal barriers restricting development of F₁ hybrids in crosses among closely related diploid species of Solanum belonging to the series Tuberosa and Commersoniana were studied to determine the early stages in development of barriers to gene exchange. Both of the parental series (species complexes) are native to South America, the Tuberosa occurring along the length of the Andes, while the Commersonia occur at lower altitudes in Argentina, Paraguay, and Bolivia. They are distinguished by a fair number of morphological characters. Following crossings between the species of each series and between species of the different series comparisons were made of the number of berries formed per attempted cross, seed per berry, percentage seed germination, and F₁ vigor and pollen fertility. Although the species used were closely enough related that all could readily be hybridized, berry and seed set following crosses between species of different series were on the average lower than they were following crosses between species of the same series. These barriers are just forming and are expressed in a varying manner, so that there were significant differences between species of the same series, and even between clones of the same species in barrier expression. The barriers of some of the species were expressed only when they were tested as female or as male parent. The inter-series F₁ seeds germinated as well as did those of the parent species and the hybrids had a normal vigor and pollen fertility.     

REPRODUCTIVE ISOLATION BARRIERS TO GENE EXCHANGE BETWEEN SOLANUM CHACOENSE AND S. COMMERSONII (SOLANACEAE)

In contrast to the general intercompatibility of diploid species of tuber-bearing potatoes of South America, Solanum chacoense Bitter and S. commersonii Dunal have well-established internal barriers to gene exchange. Pollen tubes of S. commersonii are inhibited in growth down the style of S. chacoense. While S. chacoense pollen tubes are not inhibited in S. commersonii stylar tissue, seed development is abnormal, resulting in production of large numbers of seeds that are not filled out. Even the apparently normal F₁ seed germinated poorly; approximately 0.1 germinable seeds were set per cross as compared with over 30 seeds per cross following intraspecific crosses. Lowland S. chacoense, including populations sympatric with S. commersonii, were not more effectively isolated from S. commersonii than were highland S. chacoense populations, over 500 km from the nearest S. commersonii. The F₁ hybrids produced were vigorous and highly fertile. Crosses between unrelated S. commersonii-chacoense F₁ hybrids produced only 1.3 germinable seeds per cross. This reduction resulted from low numbers of berries set, few seeds per berry, and low seed germination. The F₂ progeny were vigorous, but while many were both male and female fertile there were indications of an increase in sterility over that of the parents and F₁'s. The data were interpreted as indicating that the isolation barriers result from polygenic differences between the parent species and are composed principally of genes that function in several of the processes of seed setting. The accumulation of genes that keep these species isolated was interpreted as not having developed as a result of selection for such isolation in the region of overlap (Wallace effect), but rather as a byproduct of divergence that occurred during their allopatric evolution.     

PATTERNS OF VARIATION AND LINKAGE DISEQUILIBRIUM IN A FIELD CRICKET HYBRID ZONE

The distribution of multilocus genotypes found within a natural hybrid zone is determined by the sample of genotypes present when the hybrid zone first formed, by subsequent patterns of genetic exchange between the hybridizing taxa, and by drift and selection within each of the hybrid zone populations. We have used anonymous nuclear DNA restriction fragment polymorphisms (RFLPs) to characterize the array of multilocus genotypes present within a well-studied hybrid zone between two eastern North American field crickets, Gryllus pennsylvanicus and Gryllus firmus. These crickets hybridize along a zone of contact that extends from New England to Virginia. Previous studies have shown that both premating and postmating barriers exist between the two cricket species, but the absence of diagnostic morphological and allozyme markers has made it difficult to assess the consequences of these barriers for genetic exchange. Analyses based on four diagnostic anonymous nuclear markers indicate that hybrid zone populations in Connecticut contain few F₁ hybrids, and that nonrandom associations persist among nuclear gene markers, between nuclear and cytoplasmic markers, and between molecular markers and morphology. Field cricket populations within the hybrid zone are not “hybrid swarms” but consist primarily of crickets that are very much like one or the other of the parental species. Despite ample opportunity for genetic exchange and evidence for introgression at some loci, the two species remain quite distinct. Such a pattern appears to be characteristic of many natural hybrid zones.     

HYBRID BREAKDOWN IN PHYSIOLOGICAL RESPONSE: A MECHANISTIC APPROACH

Data comparing the physiological response to hyperosmotic stress in individual copepods (Tigriopus californicus) from natural populations and laboratory hybrids are presented. While individuals from two genetically differentiated natural populations and F₁ interpopulation hybrids showed only minor differences in patterns of free amino acid (FAA) accumulation during hyperosmotic stress, patterns of FAA synthesis were highly variable among the F₂ hybrids. Isofemale lines initiated from later hybrid generations (F₇-F₁₀) exhibited substantial inter-line variance in FAA accumulation. This increased variance in physiological response appears unrelated to allozyme polymorphisms at two loci encoding FAA-metabolizing enzymes and appears to result from other, unidentified polymorphisms in the proline synthetic pathway.     

Geographical variation in postzygotic isolation and its genetic basis within and between two Mimulus species

The aim of this study is to investigate the evolution of intrinsic postzygotic isolation within and between populations of Mimulus guttatus and Mimulus nasutus. We made 17 intraspecific and interspecific crosses, across a wide geographical scale. We examined the seed germination success and pollen fertility of reciprocal F₁ and F₂ hybrids and their pure-species parents, and used biometrical genetic tests to distinguish among alternative models of inheritance. Hybrid seed inviability was sporadic in both interspecific and intraspecific crosses. For several crosses, Dobzhansky–Muller incompatibilities involving nuclear genes were implicated, while two interspecific crosses revealed evidence of cytonuclear interactions. Reduced hybrid pollen fertility was found to be greatly influenced by Dobzhansky–Muller incompatibilities in five out of six intraspecific crosses and nine out of 11 interspecific crosses. Cytonuclear incompatibilities reduced hybrid fitness in only one intraspecific and one interspecific cross. This study suggests that intrinsic postzygotic isolation is common in hybrids between these Mimulus species, yet the particular hybrid incompatibilities responsible for effecting this isolation differ among the populations tested. Hence, we conclude that they evolve and spread only at the local scale.     

DEVELOPMENTAL STABILITY,FITNESS, AND TRAIT SIZE IN LABORATORY HYBRIDS BETWEEN EUROPEAN SUBSPECIES OF THE HOUSE MOUSE

The effects of hybridization on developmental stability and size of tooth characters were investigated in intersubspecific crosses between random-bred wild strains of the house mouse (Mus musculus domesticus and M. m. musculus). Fluctuating asymmetry (FA) and trait size were compared within and between parental, F₁, backcross, and F₂ hybrid groups. The relationship between FA and reproductive fitness within the F₁ hybrids was also studied. The results indicated that both FA and character size levels differed significantly between the two subspecies. The F₁ hybrids and the recombined groups (backcrosses and F₂ hybrids) showed heterosis for both parameters. No significant differences in the FA of fertile and sterile F₁ hybrid individuals were found. Comparison of the FA levels obtained in this study with those found in wild populations from the hybrid zone in Denmark showed that the levels of FA were lower in laboratory-bred samples than in the wild populations. This study provides further evidence that, in hybrids, the developmental processes underlying most of the morphological traits we studied benefit from a heterotic effect, despite the genomic incompatibilities between the two European house mice revealed by previous genetical and parasitological studies.     

Cytogenetic studies of the F1 hybrids of Capsicum annuum with C. chinense and C. baccatum

Summary Partially sterile interspecific hybrids were obtained between C. annuum var. cerasiformis and C. chinense var. mishme (H₁), and C. annuum var. cerasiformis and C. baccatum var. pendulum (H₂). Morphologically the F₁ hybrids were intermediate between the corresponding parents. Meiosis was irregular in the two F₁ hybrids. Cytological analysis of the two F₁ hybrids revealed that the genome of C. annuum differs from C. chinense by two translocations and some minor structural alterations and from C. baccatum by two translocations, a single inversion and some minor structural alterations. Isolation barriers such as hybrid inviability, weakness and hybrid breakdown in the H₁ hybrid and, inaddition, desynapsis in the H₂, were operative in these taxa. The differences between the present findings and those reported earlier on the two F₁hybrids were attributed to differences in the genetic architecture of the taxa employed in hybridization.     

Hybridization studies in the genusSilene sectt.Siphonomorpha andAuriculatae (Caryophyllaceae)

Ten species in the genusSilene sectt.Siphonomorpha andAuriculatae were crossed artificially involving 612 crosses to test inter- and infraspecific, intervarietal and intersectional crossability. In sect.Siphonomorpha all interspecific crosses (between diploids) failed due to cross- or seed-incompatibility; however, intervarietal crosses betweenS. gigantea var.gigantea andS. gigantea var.incana produced hybrids. In sect.Auriculatae hybrids were produced betweenS. vallesia andS. boryi, both tetraploids, but crosses between these and the diploid species were unsuccessful. The delimitation and status of the species in both sections was supported by the crossing results.     

REPRODUCTIVE BIOLOGY OF ERYTHRONIUM GRANDIFLORUM VARIETIES GRANDIFLORUM AND CANDIDUM (LILIACEAE)

This four-year (1982–1986) study compared the pollination biology and cytology of a relatively rare taxon, Erythronium grandiflorum Pursh var. candidum (Piper) Abrams, with that of a geographically widespread sister taxon, E. grandiflorum var. grandiflorum, in the Pacific Northwest. Breeding studies were used to study the reproductive systems of the taxa, to test for interfertility between them, and to seek abnormalities in the breeding system of the rare taxon that might account for its relative rarity. Chromosomes were counted in both taxa and in hybrids from artificial crosses. Ranges of the two taxa were determined from field studies and herbarium specimens. Although the ranges of the taxa overlap in part, populations are usually allopatric. Phenological factors are probably minor in preventing cross-pollination between the taxa. Both taxa are protogynous obligate outcrossers pollinated by bees. Seed production apparently is not pollinator-limited. The two taxa are interfertile only when individuals from allopatric populations are artificially crossed, and thus should be considered two biological species. When the taxa overlap in floral phenology in sympatric populations, they exhibit reciprocal incompatibility. Such incompatibility between closely related perennial species is unusual. It may have arisen via natural selection in mixed populations. Alternatively, if populations of each taxon vary with regard to their incompatibility alleles, they may have acquired their fertility barriers by chance before populations of the two taxa came into contact with each other.     

Cross-breeding studies on the cassava green spider miteMononychellus sp. (Acari: Tetranychidae) in East Africa

Crossbreeding experiments were carried out using six populations of the cassava green spider mite originating from different locations in Kenya and Uganda. In all cross-combinations made, an F₁, F₂ and F₃ generation was obtained. A slight increase in the egg mortality in the F₂ points to the fact that a partial hybrid sterility occurs in some of the cross-combinations. No indications for the existence of more severe reproductive barriers were found.Since the populations hybridize and show free gene exchange it is concluded that all populations are conspecific and belong toMononychellus progresivus Doreste.     

SEED FERTILITY IN TRAGOPOGON HYBRIDS (COMPOSITAE)

The seed fertility of 172 interspecific and 32 intraspecific F₁ hybrids in the genus Tragopogon was determined. Intrageneric barriers to interbreeding were variable; some were weak and others quite strong. Maternal influences on fertility were found in 18 pairs of reciprocal interspecific crosses. It was postulated that cytoplasmic and nuclear factors interacted to determine the level of seed fertility. Although F₁'s were generally sterile or semi-sterile, the proportion of fertile seeds produced in most crosses increased in the F₂ generation. In two cases, however, a significant decrease was observed.     

ARTIFICIAL HYBRIDIZATION IN HAWAIIAN WIKSTROEMIA (THYMELAEACEAE)

Crosses were performed within and between 19 Hawaiian populations of the mostly dioecious woody shrub Wikstroemia. These populations are from a variety of habitats on the six major islands and are morphologically diverse. The populations are referrable to eight of the 12 species recognized by Peterson. Almost all hybrid combinations produced seeds, and seed-set was not significantly different among three categories of hybrids (within population, between populations on the same island, and between populations on different islands). F₁s from many of the crosses were vigorous and fertile, leading to the conclusion that these populations are not reproductively isolated from each other and the assumption that they are genetically very similar. Natural hybridization was difficult to detect because the species described by Peterson intergrade morphologically and ecologically. The crossability and interfertility of populations of Wikstroemia may be explained by one or more of the following hypotheses: 1) recency of colonization, 2) the founder effect, 3) lack of selection for barriers to interbreeding, and 4) gross morphological changes without associated large genetic changes.     

VARIATION IN CHROMATOGRAPHIC PATTERNS IN THE TRAGOPOGON DUBIUS-PRATENSIS-PORRIFOLIUS COMPLEX (COMPOSITAE)

Natural populations of the diploid species, Tragopogon dubius, T. pratensis and T. porrifolius, the F₁ hybrids of these species, and the two amphidiploids, T. mirus and T. miscellus, were obtained from the same localities used for previous morphological and cytological studies of the evolutionary relationships of this complex. Inter- and intra-populational comparisons were made utilizing 2-dimensional chromatograms of these taxa. Members of the complex shared a group of 18 flavonoid-type pattern components, no one of which was completely species specific. Therefore, species patterns were expressed for a population rather than an individual with pattern components being expressed as a frequency. Patterns for F₁ hybrids and amphidiploids, expressed in the same manner, were relatable to the parental species. Diploid species and F₁ hybrid patterns were the most variable in areas of greatest sympatry, indicating considerable genetic interchange between species. The distribution of specific components in certain populations was interpreted as chemical introgression. Evidence was presented for separate origins of T. mirus in two localities. The results confirm previous interpretations of the evolutionary relationships in this complex, and when compared to those obtained for Baptisia, they implicate the different breeding systems as important factors in establishing the kind of variation observed.