Synchronous allelic expression at the glucosephosphate isomerase A and B loci in interspecific sunfish hybrids

Allelic isozymes of glucosephosphate isomerase at the Gpi-A and -B loci were separated by starch gel electrophoresis in the warmouth (Lepomis gulosus) and green sunfish (L. cyanellus). The specific tissue distributions and developmental expressions of the GPI-A₂, -AB, and -B₂ isozymes were not different between these two species. The synchrony of allelic expression in normal intraspecific sunfish crosses was demonstrated by means of an electrophoretic variant at the Gpi-B locus. In embryos formed from warmouth × green sunfish hybrid crosses, the paternal GPI-A₂ isozymes were first expressed at the same time in both reciprocal hybrids, at 21–25 hr after fertilization. The maternal and paternal GPI-B subunits were synchronously expressed in reciprocal hybrids just prior to hatching. The parental allelic isozymes at both loci showed codominant expression in all tissues of the mature F₁ hybrids. These results are consistent with the absence of allelic asynchrony and inhibition in interspecific hybrids formed from more evolutionarily related species.     

Changes in DNA-methylation during zygotic embryogenesis in interspecific hybrids of beans (<Emphasis Type="Italic">Phaseolus</Emphasis> ssp.)

Hybrid embryos resulting from crosses between Phaseolus species often fail to reach maturity and some combinations frequently abort at early developmental stages. The genetic or molecular basis for these consistent developmental defects is at present not clear. However, an extremely complex genetic system, thought to be caused by major epigenetic changes associated with gene expression changes, has been shown to be active in plant species. We have investigated DNA methylation in two interspecific hybrids, Phaseolus vulgaris × Phaseolus coccineus and its reciprocal crosses, using methylation sensitive amplification polymorphism (MSAP). The potential use of MSAP for detecting methylation variation during embryogenesis in interspecific hybrids is discussed. Significant differences in the DNA methylation patterns were observed in abortive (interspecific hybrids) and non abortive (parental) genotypes. Taken together, our results strongly suggest that generalized alterations in DNA methylation profiles could play a causative role in early interspecific embryo abortion in vivo. A considerable change in the methylation pattern during embryogenesis could be involved in the disruption of the regulation or maintenance of the embryogenesis process of Phaseolus interspecific hybrids. The results also support the earlier hypothesis that DNA methylation is critical for the regulation of plant embryogenesis and gene expression.     

Chloroplast DNA inheritance in theStellaria longipes complex (Caryophyllaceae)

Inheritance of chloroplast DNA (cpDNA) was examined in F₁ progenies derived from three crosses and three corresponding reciprocal crosses betweenStellaria porsildii andS. longifolia. Chloroplast DNA restriction fragments were analyzed using methods of nonradioactive digoxigenin-11-dUTP labeling and chemiluminescent detection with Lumi-Phos 530. Distinct interspecific restriction fragment polymorphisms were identified and used to demonstrate the mode of cpDNA inheritance. Mode of cpDNA inheritance differed among crosses. Two crosses in whichS. porsildii, SP2920-21, was the maternal parent exhibited three different types of plastids, maternal, paternal and biparental, among the F₁ hybrids, suggesting a biparental cpDNA inheritance and plastid sorting-out inStellaria.     

Dwarfism and male sterility in interspecific hybrids of Epilobium

Summary Reciprocal differences for male sterility, dwarfism and morphological traits have been studied in intra- and interspecific crosses of five Epilobium species. Male sterility occurred in two interspecific hybrids with E. montanum as the male parent while dwarfism has been found to varying degrees in three interspecific crosses with E. watsonni. In contrast to transient differences in plant height and leaf morphology in reciprocal hybrids of the cross between E. hirsutum and E. parviflorum, male sterility and dwarfism persistently occur as reciprocally different traits which may be influenced by determinants of the cytoplasm. The molecular characterization of the plastid DNA of the parental lines and the F₁ hybrids indicate that the plastome of male sterile and dwarf plants is identical to that of the female parents. Furthermore, in spite of these developmental disturbances, the expression of plastid genes coding for polypeptides of thylakoid-membrane complexes is unchanged. Thus, it seems unlikely that the genetic compartement of the plastids is responsible for the expression of the male sterile or the dwarfed phenotype.     

Genetic control of seed proteins in wheat

Summary Electrophoretic profiles of crude protein extracts from seed of F₁ hybrids and reciprocal crosses among diploid, tetraploid and hexaploid wheats were compared with those of their respective parental species. The electrophoretic patterns within each of three pairs of reciprocal crosses, T.boeoticum X T.urartu, T.monococcun X T. urartu and T.dicoccum X T. araraticum, were different from one another but were identical with those of their respective maternal parents. Protein bands characteristic of the paternal parents were missing in F₁ hybrid seed suggesting that the major seed proteins in wheat were presumably regulated by genotype of the maternal parent rather than by the seed genotype. However, in another three pairs of reciprocal crosses, T.boeoticum X T. durum, T.dicoccum X T.aestivum and T. zhukovskyi x T. aestivum, protein bands attributable to the paternal parents were present in the F₁ hybrid seeds indicating that the seed proteins were not always exclusively regulated by the maternal genotype. The expression of paternal genomes is presumably determined by dosage and genetic affinity of the maternal and paternal genomes in the hybrid endosperm. The maternal regulation of seed protein content is probably accomplished through the maternal control over seed size. The seed protein quality may, however, depend upon the extent of expression of the paternal genome.     

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 Genetics of Fishes: Isozymic Analyses of Differential Gene Expression

SYNOPSIS. The fundamental evolutionary position of the fishesand the plasticity of their gene expression render them wellsuited for developmental genetic analyses. Isozymes, encodedin related loci, have proven to be effective probes of differentialgene expression in the fishes. The advanced bony fishes exhibita higher temporal and spatial specificity of isozyme locus expressionthan the primitive fishes. Some isozyme loci are limited intheir expression to specific developmental periods such as hatching.The more phylogenetically recently derived loci have a morerestricted tissue expression, and tend to be expressed laterin development than more ancient loci exhibiting a more generalizedtissue expression. More closely related isozyme loci tend tooverlap more in their cellular expressions than do more distantlyrelated loci. The role of gene and genome relatedness in regulatinggene expression also has been studied by the investigation ofpreferential allele expression in interspecific hybrids. Theschedules of gene expression are often perturbed in interspecificfish hybrids. A progressive increase in the severity of developmentalabnormalities, at the morphological and enzymatic levels, occursduring embryogenesis of hybrids formed from progressively moredistantly related species. In most instances of allelic repression,the paternal allele is preferentially inhibited but occasionallythe maternal allele is selectively repressed. A non-reciprocaldevelopmental success is often observed for hybrid embryos derivedfrom reciprocal crosses. These aberrant nucleocytoplasmic interactionshave been related to models of gene regulation and the evolutionarydivergence of gene regulatory mechanisms.     

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.     

Embryo development in reciprocal crosses of Phaseolus vulgaris L. and P. coccineus Lam.

Summary Embryo development was examined in reciprocal crosses of Phaseolus vulgaris cv. Great Northern and P. coccineus cv. Scarlet Runner. The formation of abnormal (shrunken and underdeveloped) embryos constituted the primary crossing barrier between the two species when P. coccineus was the female parent. Plants of P. coccineus X P. vulgaris were obtained by embryo culture. Although the P. vulgaris X P. coccineus cross resulted in normal seed development, the fertility of the resulting hybrids was much lower (27%) than that of the reciprocal hybrids (81%). Three classes of F₂ embryos, normal, shrunken, and underdeveloped were formed on reciprocal F₁s and the frequencies did not differ between reciprocal populations. Thus, the interactions between embryo and endosperm and/or maternal parent rather than cytoplasmic-nuclear effects seem to be important in the determination of the extent of embryo growth. The examination of pollen fertility of F₂ plants and the development of F₂ and F₃ embryos suggests that the formation of abnormal embryos and reduced male fertility are independent events. The P. vulgaris — P. coccineus crosses may be useful in studying the possible involvement of interspecific differences in hormonal metabolism in the development of hybrid embryos.     

Demographic aspects of interspecific hybridization

Summary A demographic study was carried out on two closely related species of the isopods Jaera (albifrons) ischiosetosa and J. (a.) albifrons and their F ₁ hybrids. The results from the 16 possible combinations of crosses have permitted an analysis of the nature of the mechanisms assuring the isolation of the species studied. Although intraspecific crosses yield an immediate success, interspecific crosses in the absence of choice of mates progress only slowly during the course of weeks. The results of both crosses between hybrids and back-crosses turn out to be intermediate between those of intra- and interspecific crosses. The hybrids of the first generation are perfectly viable and their survival curves are identical to those of the parents. The fertility of parents in intra- and interspecific crosses is comparable, with the exception of the fragility of female descendants (heterogametic sex) in one direction of crossing. The fertility of the F ₁ hybrids, however, crossed either among themselves or with their parents, is quite noticeably decreased: the time needed to double the size of the population is 2.5 times longer for the hybrids than for the parents. This hybrid breakdown completes the pre-fertilization isolating mechanisms: partial ecological isolation, and especially ethological isolation, is practically total when a choice of mates exists. The two species studied, for which demographic parameters are quite close, were raised together for ten generations and yielded only exceptional hybrids with a frequency which does not exceed that found under natural conditions.     

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.     

INTERSPECIFIC INCOMPATIBILITY IN SOLANUM SPECIES

Pandey , K.K. (Crop Res. Div., D.S. & I.R., Lincoln, Christchurch, New Zealand.) Interspecific incompatibility in Solanum species. Amer. Jour. Bot. 49(8): 874–882. Illus. 1962.—A diallel cross involving 11 self-incompatible and 3 self-compatible species of Solanum was made to study the genetic basis of interspecific incompatibility. Interspecific incompatibility was not limited to crosses in which a self-compatible species was used as the male parent onto a self-incompatible species (unilateral incompatibility). A number of crosses between self-incompatible species were incompatible. In one cross, Q vernei X verrucosum, a self-compatible species was successful as a pollen parent with a self-incompatible species. Unlike other hybrids between self-compatible and self-incompatible species, which are self-incompatible, these F₁ hybrids were self-fertile, and cross-fertile among themselves and with both parents. The self-fertile S. polyadenium was cross-incompatible as a female as well as a male parent with all other species. It is suggested that the unilateral incompatibility is a property of the allele S_C which originated as a consequence of one kind of breakdown of the S_I gene; the S_C allele produces “bare” pollen growth substances which are inactivated in an incompatible style. It is proposed that the failure of the principle of unilateral interspecific incompatibility in solanaceous species may be due to the action of alleles at the second incompatibility locus revealed in certain Mexican species. It is assumed that the South American species are selected intraspecifically only for the action of S alleles but that in certain interspecific crosses and rarely in intraspecific crosses the alleles at the second locus may be expressed, thus interfering with the usual action of S alleles. The F₁ hybrids Q verrucosum (self-fertile) X simplicifolium (self-sterile) were self-incompatible at the tetraploid as well as the diploid level, and their cross-compatibility behavior was consistent with the expected activity of the S_C and S_I alleles of the 2 parents respectively.     

A study of wing morphology and fluctuating asymmetry in interspecific hybrids between Drosophila buzzatii and D. koepferae

In this work we investigate the effect of interspecific hybridization on wing morphology using geometric morphometrics in the cactophilic sibling species D. buzzatii and D. koepferae. Wing morphology in F₁ hybrids exhibited an important degree of phenotypic plasticity and differs significantly from both parental species. However, the pattern of morphological variation between hybrids and the parental strains varied between wing size and wing shape, across rearing media, sexes, and crosses, suggesting a complex genetic architecture underlying divergence in wing morphology. Even though there was significant fluctuating asymmetry for both, wing size and shape in F₁ hybrids and both parental species, there was no evidence of an increased degree of fluctuating asymmetry in hybrids as compared to parental species. These results are interpreted in terms of developmental stability as a function of a balance between levels of heterozygosity and the disruption of coadaptation as an indirect consequence of genomic divergence.     

Occurrence of genetic tumours in Triticum interspecies hybrids

Summary F₁ interspecific hybrids involving nine tetraploid Triticum species were studied. Some developed leaf tumours at the seedling stage. Tumorous hybrids were restricted to crosses involving either T. timopheevi or T. araraticum as one parent. The hybrids from the rest of the crosses, including those of T. timopheevi × T. araraticum, were non-tumorous. Genetically divergent and non-integrated parental species appeared to be inducing spontaneous tumour formation in their hybrids.     

Molecular analysis of Arachis interspecific hybrids

Incorporation of genetic resistance against several biotic stresses that plague cultivated peanut, Arachis hypogaea (2n=4x=40), is an ideal option to develop disease resistant and ecologically safe peanut varieties. The primary gene pool of peanut contains many diploid wild species (2n=2x=20) of Arachis, which have high levels of disease and insect resistances. However, transfer of resistant genes from these species into A. hypogaea is difficult due to ploidy level differences and genomic incompatibilities. This study was conducted to monitor alien germplasm transmission, using Random Amplified Polymorphic DNA (RAPD) markers, from two diploid wild species, A. cardenasii and A. batizocoi, into A. hypogaea. Triploid interspecific hybrids were produced by crossing two A. hypogaea cultivars (NC 6 and Argentine) with the two species and by colchicine-treating vegetative meristems, fertility was restored at the hexaploid (C_o) level in the four hybrids. Hexaploids were allowed to self-pollinate for four generations, each referred to as a cycle (C₁, C₂, C_3, and C₄). At each cycle, a backcross was made with the respective A. hypogaea cultivar as the maternal parent and only lineages tracing back to a single hexaploid hybrid were used for RAPD analysis. Analysis of mapped, species-specific RAPD markers in BC₁F₁ to BC₁F₃ hybrids indicated that alien germplasm retention decreased every generation of inbreeding, especially in Argentine and in A. batizocoi crosses. A similar trend was also observed for every cycle in BC₁F₂ and BC₁F₃ families, possibly, due to the loss of alien chromosomes following selfing of hexaploids. RAPD marker analysis of 40–chromosome interspecific hybrid derivatives from the four crosses supported previous reports that reciprocal recombination and/or translocations are the predominant mechanisms for exchange of chromosomal segments. No evidence was found for preferential transfer of alien chromosomal regions to specific linkage groups. The implications for developing disease resistant peanut breeding lines are discussed in light of these findings.     

Exogenous selection rather than cytonuclear incompatibilities shapes asymmetrical fitness of reciprocal Arabidopsis hybrids

Reciprocal crosses between species often display an asymmetry in the fitness of F₁ hybrids. This pattern, referred to as isolation asymmetry or Darwin's corollary to Haldane's rule, is a general feature of reproductive isolation in plants, yet factors determining its magnitude and direction remain unclear. We evaluated reciprocal species crosses between two naturally hybridizing diploid species of Arabidopsis to assess the degree of isolation asymmetry at different postmating life stages. We found that pollen from Arabidopsis arenosa will usually fertilize ovules from Arabidopsis lyrata; the reverse receptivity being less complete. Maternal A. lyrata parents set more F₁ hybrid seed, but germinate at lower frequency, reversing the asymmetry. As predicted by theory, A. lyrata (the maternal parent with lower seed viability in crosses) exhibited accelerated chloroplast evolution, indicating that cytonuclear incompatibilities may play a role in reproductive isolation. However, this direction of asymmetrical reproductive isolation is not replicated in natural suture zones, where delayed hybrid breakdown of fertility at later developmental stages, or later‐acting selection against A. arenosa maternal hybrids (unrelated to hybrid fertility, e.g., substrate adaptation) may be responsible for an excess of A. lyrata maternal hybrids. Exogenous selection rather than cytonuclear incompatibilities thus shapes the asymmetrical postmating isolation in nature.     

CROSSING RELATIONSHIPS AMONG DIPLOID SPECIES OF THE SOLANUM NIGRUM COMPLEX IN NORTH AMERICA

Crosses were made in all combinations of the six diploid species of the Solarium nigrum complex that occur in North America. Some interspecific pollinations failed to yield viable seed; successful crosses gave rise to moderately to highly sterile F₁ hybrids. Results of interspecific crosses suggested phylogenetic relationships that were not completely in accord with those suggested by morphology. Interspecific crosses also gave varied results. All interpopulational crosses within S. interius and S. sarachoides produced fully fertile hybrids. In contrast, hybrids within S. americanum and S. douglasii varied from fully fertile to almost completely sterile. Populations of S. pseudogracile could be divided into two groups which are geographically separated but not morphologically differentiated. Fully fertile hybrids resulted from crosses within a group, whereas crosses between groups gave hybrids with reduced fertility. Four crossing groups were observed within 5. nodiflorum; three of the parental groups are sympatric and are morphologically differentiated. Although hybrid sterility in interspecific crosses is sometimes used to support delimitation of species, the presence of sterility in intraspecific crosses suggests that such an interpretation is unwarranted for the S. nigrum complex. Hybrid sterility, therefore, is not considered to have special taxonomic significance in this complex.     

Relative developmental success of interspecific Lepomis hybrids as an estimate of gene regulatory divergence between species

The developmental success of interspecific Lepomis hybrids is used as an index of gene regulatory divergence between the green sunfish, L. cyanellus, and each of three other parental species, longear sunfish, L. megalotis, warmouth, L. gulosus, and bluegill, L. macrochirus. This gene regulatory divergence is compared to the degree of structural gene divergence among these four species (genetic distance [Nei, '78], D, ranged from 0.206 to 0.586). The developmental success of the hybrid embryos at the level of morphogenesis was higher than expected from the genetic distance between the parental species. The rates of morphogenesis of the hybrid embryos were the same as that for the green sunfish embryos. The percentage of embryos that hatched was relatively high in all crosses. However, two of the hybrid crosses resulted in enhanced percentages of hatched embryos. Slight increases in the extent of morphological abnormalities were observed in hybrids from crosses between more distantly related parental species. The schedules and levels of enzyme locus expression of the hybrids, assessed spectrophotometrically and electrophoretically for nine enzyme systems (encoded in a total of 14 loci), were different from each other and from those of the green sunfish embryos. Alterations in the time of first enzyme appearance and in the time of first increase in enzyme activity in the developing hybrid embryos were not correlated with genetic distance between parental species. However, the extents of alteration of enzyme activities over the entire period of hybrid embryogenesis were correlated with the genetic distance. We attribute the morphological and molecular anomalies observed in the hybrids to gene regulatory incompatibilities between species. Although the exact number of mutational differences and their relative developmental impacts are not known, some inferences can be drawn about the degree of divergence in gene regulation between species. It appears that an uncoupling of the rates of structural and regulatory gene evolution can occur between species of some taxa, an observation that has implications for the roles of gene regulatory differences in organismic evolution.     

Analysis of production,early growth and survival ofClarias gariepinus (Burchell),Heterobranchus longifilis (Val.) (Pisces: Clariidae) and their F1 hybrids in ponds

Clarias gariepinus andHeterobranchus longifilis were successfully hybridized to produce reciprocal F₁ progeny during a two-year study.C. gariepinus produced significantly heavier spawn thanH. longifilis during the same period. Fertilized eggs were incubated in aerated static-water in fibre-glass troughs, and hatching rates were not found to be significantly different (P<0.05) between any two of the four catfish groups produced. Eight-day old fry of each group were reared in fertilized nursery ponds, and F₁ hybrids fromH. longifilis maternal parent had the best growth and survival rates at the end of 18 days. The reciprocal cross showed intermediate growth rate between both parents. At harvest, differential growth was less pronounced among both hybrids than among the parents.     

Genetic evidence for natural hybridization and apparent introgression between freshwater snail species (Viviparus ater and V. contectus)

We examined six allozyme markers and shell morphology to study gene flow between the naturally sympatric freshwater snail species Viviparus ater and V. contectus in Lake Garda, Italy, using offspring from experimental crosses and snails collected from natural populations. Hybrid offspring obtained from experimental crosses and hybrids collected from natural populations were heterozygous at four or five out of five diagnostic loci. Shell morphology was a poor predictor of the hybrid status of individual snails. Backcrossed offspring from experimentally bred hybrids and either of the parental species morphologically resembled the parental species. There was no evidence of segregation distortion at the six loci. Linkage analysis revealed one pair of linked loci (GPI and PNP). These cross experiments indicated a Mendelian type hybrid system in which gene introgression may occur. F₁ hybrids were found at four out of six sampling sites of Lake Garda, Italy. Local frequencies of F₁ hybrids ranged from 0% to 1.6% (estimated average = 0.74%) of the total population including both species. Alleles typical of V. ater were found at low frequencies in V. contectus at all six sampling sites. Alleles typical of V. contectus were found at low frequencies in V. ater at three out of six sampling sites. This is consistent with the hypothesis of introgression in both directions.