Development and characterization of microsatellite markers for Prosopis chilensis and Prosopis flexuosa and cross‐species amplification

Prosopis chilensis and Prosopis flexuosa (Fabaceae) are closely related hardwood arboreal species that are widely distributed in the arid regions of Argentina. The development of highly polymorphic markers, such as microsatellites, is desirable for genetic studies of these species. Here, we present the development and characterization of six polymorphic microsatellite markers in P. chilensis and P. flexuosa. These markers showed a polymorphism information content between 0.14 and 0.85 and the number of alleles varied from two to 13 considering both species. All markers revealed a broad cross‐species affinity when tested in seven other Prosopis species. All primers amplified in at least five species.     

Transgressive character expression in hybrid zones between the native invasives Tithonia tubaeformis and Tithonia rotundifolia (Asteraceae) in Mexico

Natural hybridization frequently promotes gene introgression among closely related species in sympatric populations, producing complex patterns of morphological variation. Therefore, a detailed understanding of the dynamics of interspecific gene flow and its morphological patterns is of widespread interest. We tested if introgressive hybridization promotes an increase in transgressive characters in comparison with the parental species. A sunflower species complex occurring in Mexico formed by two native invasive species, Tithonia tubaeformis and Tithonia rotundifolia, was analyzed using 46 morphological characters (leaf, flower and fruit) in five hybrid zones (N = 150 individuals) and two pure sites for each parental species (N = 80 individuals). In general, T. tubaeformis differed significantly from T. rotundifolia in all the examined characters, except six foliar and one inflorescence character. Morphological characters support the hypothesis of hybridization in this complex, even though both species remain morphological distinct in mixed stands. Individual hybrids appear to be a mosaic of parent-like (24.8 % of traits), intermediate (26.1 %) and transgressive (37.8 %) phenotypes (the remaining 11.3 % of the traits did not differ significantly from both parental species). Our results suggest that individuals from the same parental species were more similar among themselves than to putative hybrids, indicating occasional hybridization with segregation in hybrid types or backcrossing to parents. Evidence indicates a unidirectional pattern of gene flow toward T. rotundifolia.     

Use of differential water sources by Prosopis flexuosa DC: a dendroecological study

In central-western Argentina, there is a pronounced water deficit gradient, from semiarid climate conditions with 500-mm rainfall/year to arid climate conditions with 80-mm rainfall/year. This climatic transition, governed by the rainfall gradient, occurs between the Arid Chaco and Monte phytogeographic regions and is evidenced by differences in vegetation type, structure, dynamics and tree growth. In turn, the availability of soil moisture, particularly access to the water table, modifies water use strategies by trees along this gradient. We analyzed how water availability, expressed as differences in accessibility to the water table, influences Prosopis flexuosa tree rings along a precipitation gradient. In this manner, we try to interpret the growth of species according to the use of differential water sources. P. flexuosa showed highly varying growth reactions (tree-ring width and hydraulic anatomic parameters) with climate, depending on the ecology of the site. Along the Arid Chaco-Monte gradient, the growth of P. flexuosa is more dependent on variations in rainfall in those areas where water depth is greater than root spread. The climate signal was hidden in those regions where the water table is accessible to the root system.     

Methodological approaches for studying the european water frog <Emphasis Type="Italic">Pelophylax esculentus</Emphasis> complex

The European water frog (Pelophylax esculentus) complex represents a unique and adequate model system for the study of interspecific hybridization and the mechanisms enabling interspecific hybrids to overcome the reproductive barriers. The difficulties in the study of individuals from the P. esculentus complex are associated with high polymorphism of morphological characters in parental species and interspecific hybrids, as well as with the presence of polyploid hybrid forms. From the discovery of the phenomenon of interspecific hybridization and the demonstration of successful reproduction of interspecific hybrids, researchers constantly searched for the methods necessary for the most accurate identification of parental species and various hybrid forms. This review describes biochemical, cytogenetic, and molecular methods and approaches used to identify individuals from the European water frog complex, as well as to analyze the genomes transferred with the gametes of hybrids. The advantages and disadvantages of these approaches are discussed. The presented methods can be used for studying other hybrid complexes of fish, amphibians, and reptiles.     

Natural Hybridization between Two Swallowtail Species Parnassius nomionand Parnassius bremeri(Lepidoptera,Papilionidae) Shown by RAPD–PCR

Genetic evidence for interspecific hybridization between Parnassius nomionand Parnassius bremeriin nature is presented. To demonstrate hybridization between these species, RAPD analysis was used. By testing 25 decamer primers, three and two diagnostic markers were revealed for P. nomionandP. bremeri, respectively. Out of 28 animals examined, 4 were shown to be interspecific hybrids. According to the distribution of diagnostic markers, the interspecific hybrids were intermediate with regard to the parental species. Ecological and biological characteristics of two butterfly species that promote their hybridization in nature are discussed.     

Morphological Variation in Wild Marmosets (Callithrix penicillata and C. geoffroyi) and Their Hybrids

Evolutionary theory and observation predict wider phenotypic variation in hybrids than parental species. Emergent phenotypic novelty in hybrids may in turn drive new adaptations or speciation by breaking parental phenotypic constraints. Primate hybridization is often documented through genetic evidence, but knowledge about the primate hybrid phenotype remains limited due to a small number of available studies on hybrid primate morphology. Here, we examine pelage and morphometric variation in two Brazilian marmoset species (Callithrix penicillata and C. geoffroyi) and their hybrids. Hybrids were sampled in an anthropogenic hybrid zone in the municipality of Viçosa, Minas Gerais state, Brazil. We analyzed hybrid facial and body pelage color variation, and compared 13 morphometric measures between hybrids and parental species. Five different hybrid facial morphotypes were observed, varying from intermediate to parental-like. Hybrid facial morphotypes were biased towards C. penicillata, suggesting that the pelage of this species may be dominant to that of C. geoffroyi in this context, and indicating that mate preference, and therefore gene flow/introgression, may be biased towards C. penicillata within the hybrid zone. Hybrid morphometric features were on average intermediate to parental species traits, but transgressive hybrids were also observed, suggesting that morphometric variation for the studied traits is consistent with Rieseberg’s complementary allele model. Finally, we observed a decoupling of facial patterning and size/shape in hybrids, relative to parent phenotypes, suggesting that an important factor driving phenotypic novelty within the Viçosa marmoset hybrid zone might be the loosening of evolutionary constraints on phenotypic trait integration.     

Interspecific and temporal variation in herbivore responses to hybrid willows

We studied herbivory of two species of willows (Salix sericea and S. eriocephala) and their interspecific hybrids to test alternative hypotheses concerning the effects of hybridization on plant resistance. Individually marked plants were identified using morphological traits in the field and random amplified polymorphic DNA (RAPD) band analysis was used to verify the genetic status of many parental and hybrid plants. The desities of 12 herbivore species on plants in the field were compared between two parents and their F₂-type hybrids. We found about equal support for the additive, dominance, and hybrid susceptibility hypotheses over 4 years. In one year, one species supported the hybrid resistance hypothesis. Guild membership was not a good predictor of similar responses of species to hybrid versus parental plants. There were marked differences in support for particular hypotheses among years for four herbivore species. This study demonstrates the diversity of responses of phytophages in response to interspecific hybridization, and indicates that year-to-year variation in relative resistance of hybrid plants can be important.     

A dynamic evolution of chromosome in subgenus Potamogeton revealed by physical mapping of rDNA loci detection

In this study, we analyzed 12 species in the subgenus Potamogeton (8 tetraploids and 2 diploids, 2 putative tetraploid hybrids) at the chromosomal level, including counting the chromosome number and physically mapping the rDNA. The extent of variation in the chromosome number and rDNA loci was determined in the Potamogeton species on both the inter- and intra-specific level. Moreover, one of the hybrid sets (P. perfoliatus ♂?×?P. wrightii ♀) was picked for performing artificial pollination for producing F1 generation adults and for sequential morphological character analysis and FISH detection. After comparing the parental species and natural hybrids from three different geographic locations, the intraspecific variations of rDNA loci were revealed in sampled plants; in addition, comparisons between artificial F1 and natural hybrids (P.?×?intortusifolius) showed a rapid change of 45S rDNA loci in response to the interspecific hybridization. We also compared our results concerning rDNA patterns with phylogenetic documents to derive complementary clues to karoytype evolution and interspecific relationships. Based on frequent hybridizations and active clonal reproduction with weak genetic selection, the rDNA chromosomal repatterning, such as the gain or loss of rDNA loci together with rDNA movements, might be one trend of chromosome evolution in this genus.     

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.     

Hybridization and transgressive exploration of colour pattern and wing morphology in Heliconius butterflies

Hybridization can generate novel phenotypes distinct from those of parental lineages, a phenomenon known as transgressive trait variation. Transgressive phenotypes might negatively or positively affect hybrid fitness, and increase available variation. Closely related species of Heliconius butterflies regularly produce hybrids in nature, and hybridization is thought to play a role in the diversification of novel wing colour patterns despite strong stabilizing selection due to interspecific mimicry. Here, we studied wing phenotypes in first‐ and second‐generation hybrids produced by controlled crosses between either two co‐mimetic species of Heliconius or between two nonmimetic species. We quantified wing size, shape and colour pattern variation and asked whether hybrids displayed transgressive wing phenotypes. Discrete traits underlain by major‐effect loci, such as the presence or absence of colour patches, generate novel phenotypes. For quantitative traits, such as wing shape or subtle colour pattern characters, hybrids only exceed the parental range in specific dimensions of the morphological space. Overall, our study addresses some of the challenges in defining and measuring phenotypic transgression for multivariate traits and our data suggest that the extent to which transgressive trait variation in hybrids contributes to phenotypic diversity depends on the complexity and the genetic architecture of the traits.     

Interspecific hybridization of plants and resistance to herbivores: hypotheses,genetics, and variable responses in a diverse herbivore community

We studied the morphology, molecular genetics, and hebivory of two species of willows (Salix sericea and S. eriocephala) and their interspecific hybrids to test four alternative hypotheses concerning the effects of hybridization on plant resistance. Individually marked plants were identified using morphological traits in the field and measurements of stipule and leaf pubescence were made and compared using Canonical Discriminant Function Analysis. DNA was extracted from the leaves of a sample of the marked plants and RAPD-PCR analysis was performed to establish the genetic status of parental and hybrid plants. RAPD band analysis generally verified the genetic status of parental plants. Hybrid plants were usually correctly identified in the field with a few exceptions. However, the hybrid plants were a heterogeneous group of plants made up of most plants that appear to be F₁s and a few plants that appear to be backcrosses to S. sericea. Morphological variables were useful for distinguishing S. sericea from S. eriocephala and hybrids, but were not as dependable in distinguishing between S. eriocephala and hybrids. We compared the densities of 11 herbivore species and the infection by a leaf rust pathogen (Melampsora sp.) on the leaves and stems of two parents and the hybrids in the field. We found support for the Additive hypothesis (3 species), the Dominance hypothesis (2 species) and the Hybrid Susceptibility hypothesis (7 species, 6 herbivores and the Melampsora rust). We found no evidence for the Hybrid Resistance hypothesis. Guild membership was not a good predictor of similar responses of species to hybrid versus parental plants. A Canonical Discriminant Function Analysis showed discrete separation of the taxa based on herbivore densities, illustrating different community structures on hybrid and parental plants. This study demonstrates the diversity of responses of phytophages in response to interspecific hybridization.     

Interspecific hybridization in the genus Hieracium s. str.: evidence for bidirectional gene flow and spontaneous allopolyploidization

Although reticulation has indisputably played an important role in the evolutionary history of the genus Hieracium s. str. (Asteraceae), convincingly documented cases of recent interspecific hybridization are very rare. Here we report combined evidence on recent hybridization between two diploid species, Hieracium alpinum and H. transsilvanicum. The hybrid origin of the plants from the Romanian Eastern Carpathians was supported by additive patterns of nuclear ribosomal DNA polymorphism (ITS), an intermediate position of hybrid plants in principal coordinate analysis based on amplified fragment length polymorphism phenotypes (AFLP), and additivity at one allozyme locus. Flow cytometric analyses and chromosome counting showed that two hybrids were diploid (2n ~ 2x ~ 18) while one was surprisingly tetraploid (2n = 4x = 36). To our knowledge, this is the first record of spontaneous polyploidization following interspecific crossing in the genus. Allozyme data, especially the presence of unbalanced heterozygosity at one locus, suggest the origin of this tetraploid via a triploid bridge with subsequent backcrossing to H. alpinum. According to PCR-RFLP analyses of the trnT-trnL intergenic spacer, all H. ×krasani hybrids examined had the H. alpinum haplotype while H. transsilvanicum served as a pollen donor. The hybrids occurred at the locality with abundant H. alpinum plants where paternal H. transsilvanicum was missing. Previously reported instances of interspecific hybridization between the same parental taxa showed an opposite direction of crossing and relative abundance of parental taxa. This suggests that the direction of hybridization might be influenced by the frequency of parental taxa at the locality.     

Genetic and morphometric markers are able to differentiate three morphotypes belonging to Section Algarobia of genus Prosopis (Leguminosae, Mimosoideae)

The section Algarobia of genus Prosopis includes promising species for reforestation and afforestation programmes in arid and semiarid regions, mainly of the Americas. Many interspecific natural hybrid combinations have been described in this group. In this paper we analysed a hybrid zone in Chaco biogeographical province in Argentina, where P. ruscifolia and P. alba overlap and hybridise producing intermediate fertile hybrid forms. Eleven morphological traits and 76 loci RAPD were analysed to determine the effect of hybridization between these species. The comparison of morphological traits among groups yielded significant or highly significant differences for all traits. Estimates of H _e in P. alba and P. ruscifolia did not differ from each other, but both showed significantly lower values than the hybrid group. The analysis of correlations between shared phenotypes and pair-wise relationships estimated from RAPD gave also strong support to the hypothesis that most of the phenotypic traits analysed have significant heritability. The analyses of population structure and clustering based on morphological and molecular data by DAPC and STRUCTURE were rather consistent and indicated that the three morphotypes studied here are differentiated with low overlapping. All results indicated that despite the occurrence of natural hybridization and introgression, interspecific gene flow would be limited by hybrid breakdown or natural selection favouring the maintenance of species integrity.     

Are linear‐leaved Potamogeton hybrids really so rare? Molecular evidence for multiple hybridizations between P. acutifolius and P. compressus in central Europe

Due to insufficient morphological differentiation, hybridization among linear‐leaved species of Potamogeton is rarely observed and recognized. Here, we applied molecular tools (sequencing of nrITS and rpl32‐trnL cpDNA intergenic spacer) to study several natural, central European populations of morphologically intermediate forms between two closely related species, P. acutifolius and P. compressus, to examine their taxonomical status and test whether they represented the variation range of the species or are results of interspecific hybridization. Both DNA regions provided distinguishing characters between putative parental species. The ITS sequences from all the morphologically intermediate specimens displayed an additive pattern combining the variation of both parental species, providing evidence for the hybrid origin of all these plants. This case study suggests that hybrids of linear‐leaved Potamogeton species are more common than usually believed, but have been difficult to identify without molecular tools.     

Daphnia Galeata × D. Longispina Hybrids in Western Norway

We describe the occurrence of D. galeata×longispina hybrids in two lakes of western Norway. Parental species and interspecific hybrids were characterised by both nuclear and mitochondrial molecular markers. In one of the populations, hybrids were shown to dominate the population over several years. A few individuals in both populations were probably not F1 hybrids, but possibly backcrosses or F2 hybrids. Most (possibly all) F1 hybrids were of D. galeata maternal origin. In addition, interspecific hybrids could be identified based on morphological characters, which were intermediate between the parental species. Interspecific hybridisation between these two species is remarkable, since they are distantly related.     

The interspecific hybrid Petunia parodii × P. inflata and its relevance to somatic hybridization in the genus Petunia

Summary Attempts at the reciprocal cross between Petunia parodii and P. inflata using standard emasculation and pollination techniques failed. Limited pollen tube growth down the style in reciprocal crosses led to reproductive isolation between the self-compatible P. parodii and self-incompatible P. inflata. The interspecific hybrid was successfully produced by bud-pollination of P. parodii with P. inflata as the male parent in 22 percent of attempts, but not in the opposite direction. In vitro pollination of P. parodii ovaries with P. inflata pollen also produced hybrids. The small size of the ovary made it technically impossible to use P. inflata as the female parent for in vitro pollination. The interspecific hybrids were intermediate, as compared to the two parents, for six of the seven plant and flower characters measured. Furthermore, the hybrids had high pollen fertility, set abundant seed upon self-pollination, and readily inter-crossed with the parental species. The results are consistent with a high degree of chromosomal homology in the parental species and with minor genetic divergency leading to reproductive isolation that is pre-zygotic in nature. Overcoming the barriers to cross-incompatibility by practical techniques resulted in fertile interspecific hybrids that segregated for parental characters. The potential value of employing the parental species in somatic hybridization experiments is discussed.Michigan Agricultural Experiment Station Journal Article No. 8404     

The influence of Prosopis canopies on understorey vegetation: Effects of landscape position

Abstract. The influence of canopy trees and shrubs on under‐storey plants is complex and context‐dependent. Canopy plants can exert positive, negative or neutral effects on production, composition and diversity of understorey plant communities, depending on local environmental conditions and position in the landscape. We studied the influence of Prosopis velutina (mesquite) on soil moisture and nitrogen availability, and understorey vegetation along a topographic gradient in the Sonoran Desert. We found significant increases in both soil moisture and N along the gradient from desert to riparian zone. In addition, P. velutina canopies had positive effects, relative to open areas, on soil moisture in the desert, and soil N in both desert and intermediate terrace. Biomass of understorey vegetation was highest and species richness was lowest in the riparian zone. Canopies had a positive effect on biomass in both desert and terrace, and a negative effect on species richness in the terrace. The effect of the canopy depended on landscape position, with desert canopies more strongly influencing soil moisture and biomass and terrace canopies more strongly influencing soil N and species richness. Individual species distributions suggested interspecific variation in response to water‐ vs. N‐availability; they strongly influence species composition at both patch and landscape position levels.     

Patterns of inheritance of mating signals in interspecific hybrids between sailfin and shortfin mollies (Poeciliidae: Poecilia: Mollienesia)

Differences in male morphology and mating behaviors are thought to confer species sexual isolation between sailfin and shortfin species of mollies. This study used interspecific crosses between the sailfin molly, P. latipinna, and the shortfin molly, P. mexicana, to investigate patterns of inheritance of morphological traits and behavioral rates of three mating behaviors in F1 hybrid males. The two parental species showed clear species differences with respect to the length of the dorsal fin and dorsal fin ray number. First generation hybrid males were intermediate between the two parental species for dorsal fin length and fin ray number, suggesting autosomal control of this trait with little effect of dominance by genes from either parental species. Parental species showed clear species differences in their rates of courtship displays. Unlike the pattern for dorsal fin morphology, F1 hybrid males showed a clear distinction in display rates with respect to the direction of the interspecific cross. Male hybrids whose sires were P. latipinna had courtship display rates that were up to three times higher than the rates of displays performed by hybrid males whose fathers were P. mexicana. The distribution of phenotypes between the parental species and that of hybrid males sired by that parental species was nearly identical. Such a pattern suggests the influence of Y-linked genes on the inheritance of courtship display rates in mollies.     

A meristic trait in Drosophila melanogaster x simulans hybrids

A quantitative trait, abdominal bristle number, is described in a number of stocks of the two sibling species Drosophila melanogaster and Drosophila simulans and in interspecific hybrids obtained from these stocks. Coefficients of variation for the trait in the parental species and the hybrids are compared, and in the case of the latter are found to fall in the same range as those of the pure species. The results are discussed in the light of previous findings on other bristle systems.