Reproductive Isolation Barriers Research Articles

Pollen competition as a reproductive isolation barrier represses transgene flow between compatible and co-flowering citrus genotypes.

Background/ObjectiveDespite potential benefits granted by genetically modified (GM) fruit trees, their release and commercialization raises concerns about their potential environmental impact, and the transfer via pollen of transgenes to cross-compatible cultivars is deemed to be the greatest source for environmental exposure. Information compiled from field trials on GM trees is essential to propose measures to minimize the transgene dispersal. We have conducted a field trial of seven consecutive years to investigate the maximum frequency of pollen-mediated crop-to-crop transgene flow in a citrus orchard, and its relation to the genetic, phenological and environmental factors involved.Methodology/Principal FindingsThree different citrus genotypes carrying the uidA (GUS) tracer marker gene (pollen donors) and a non-GM self-incompatible contiguous citrus genotype (recipient) were used in conditions allowing natural entomophilous pollination to occur. The examination of 603 to 2990 seeds per year showed unexpectedly low frequencies (0.17–2.86%) of transgene flow. Paternity analyses of the progeny of subsets of recipient plants using 10 microsatellite (SSR) loci demonstrated a higher mating competence of trees from another non-GM pollen source population that greatly limited the mating chance of the contiguous cross-compatible and flowering-synchronized transgenic pollen source. This mating superiority could be explained by a much higher pollen competition capacity of the non-GM genotypes, as was confirmed through mixed-hand pollinations.Conclusions/SignificancePollen competition strongly contributed to transgene confinement. Based on this finding, suitable isolation measures are proposed for the first time to prevent transgene outflow between contiguous plantings of citrus types that may be extendible to other entomophilous transgenic fruit tree species.

Genetic Architecture of a Reinforced, Postmating, Reproductive Isolation Barrier between Neurospora Species Indicates Evolution via Natural Selection

A role for natural selection in reinforcing premating barriers is recognized, but selection for reinforcement of postmating barriers remains controversial. Organisms lacking evolvable premating barriers can theoretically reinforce postmating isolation, but only under restrictive conditions: parental investment in hybrid progeny must inhibit subsequent reproduction, and selected postmating barriers must restore parents' capacity to reproduce successfully. We show that reinforced postmating isolation markedly increases maternal fitness in the fungus Neurospora crassa, and we detect the evolutionary genetic signature of natural selection by quantitative trait locus (QTL) analysis of the reinforced barrier. Hybrid progeny of N. crassa and N. intermedia are highly inviable. Fertilization by local N. intermedia results in early abortion of hybrid fruitbodies, and we show that abortion is adaptive because only aborted maternal colonies remain fully receptive to future reproduction. In the first QTL analysis of postmating reinforcement in microbial eukaryotes, we identify 11 loci for abortive hybrid fruitbody development, including three major QTLs that together explain 30% of trait variance. One of the major QTLs and six QTLs of lesser effect are found on the mating-type determining chromosome of Neurospora. Several reinforcement QTLs are flanked by genetic markers showing either segregation distortion or non-random associations with alleles at other loci in a cross between N. crassa of different clades, suggesting that the loci also are associated with local effects on same-species reproduction. Statistical analysis of the allelic effects distribution for abortive hybrid fruitbody development indicates its evolution occurred under positive selection. Our results strongly support a role for natural selection in the evolution of reinforced postmating isolation in N. crassa.

THE CONCEPT OF EFFECTIVE RECOMBINATION RATE AND ITS APPLICATION IN SPECIATION THEORY

The goal of this study is to develop a unifying theoretical framework to quantify the strength of reproductive isolation. We propose the use of the "effective recombination rate," which measures how fast associations of genes are broken by interlocus recombination. Applying the well-established theory of the effective migration rate, we derive two techniques to investigate the effective recombination rate in models of speciation: the weak migration approximation for parapatric scenarios and the weak recombination approximation for sympatric scenarios. We illustrate the use of these two methods by two examples each: (1) single-locus genetic incompatibility and (2) two-locus genetic incompatibility for the first method, and (3) assortative mating and (4) assortative mating combined with disruptive selection for the second method. An advantage of the effective recombination rate over previous approaches is that it integrates gene flow in both directions into a single index measuring the strength of isolation. This enables straightforward comparisons of speciation scenarios with the same or different geographic histories. The method also allows us to evaluate the relative contributions of F2 hybrid deficiency or linkage between multiple barriers in reproductive isolation.

Reinforced postmating reproductive isolation barriers inNeurospora, an Ascomycete microfungus

Maladaptive hybridization promotes reinforcement, selection for stringent reproductive isolation barriers during speciation. Reinforcement is suspected when barriers between sympatric populations are stronger than allopatric barriers, and particularly when stronger barriers evolve in the species and sex suffering the greatest costs of hybridization. Canonically, reinforcement involves premating barriers. Selection for postmating barriers is controversial, but theoretically possible. We examined geographical patterns in reproductive isolation barriers between Neurospora crassa and Neurospora intermedia, fungi with pheromone-mediated mate recognition and maternal care. We find that isolation is stronger between sympatric populations than allopatric populations, and stronger barriers are associated with the species (N. crassa) and mating role (maternal) suffering the greater costs of hybridization. Notably, reinforced isolation involves a postmating barrier, abortion of fruitbodies. We hypothesize that fruitbody abortion is selectively advantageous if it increases the likelihood that maternal Neurospora individuals successfully mate conspecifically after maladaptive hybrid fertilization.

Specialized cells tag sexual and species identity in Drosophila melanogaster

Social interactions depend on individuals recognizing each other, and in this context many organisms use chemical signals to indicate species and sex. Cuticular hydrocarbon signals are used by insects, including Drosophila melanogaster, to distinguish conspecific individuals from others. These chemicals also contribute to intraspecific courtship and mating interactions. However, the possibility that sex and species identification are linked by common chemical signalling mechanisms has not been formally tested. Here we provide direct evidence that a single compound is used to communicate female identity among D. melanogaster, and to define a reproductive isolation barrier between D. melanogaster and sibling species. A transgenic manipulation eliminated cuticular hydrocarbons by ablating the oenocytes, specialized cells required for the expression of these chemical signals. The resulting oenocyte-less (oe(-)) females elicited the normal repertoire of courtship behaviours from males, but were actually preferred over wild-type females by courting males. In addition, wild-type males attempted to copulate with oe(-) males. Thus, flies lacking hydrocarbons are a sexual hyperstimulus. Treatment of virgin females with the aversive male pheromone cis-vaccenyl acetate (cVA) significantly delayed mating of oe(-) females compared to wild-type females. This difference was eliminated when oe(-) females were treated with a blend of cVA and the female aphrodisiac (7Z,11Z)-heptacosadiene (7,11-HD), showing that female aphrodisiac compounds can attenuate the effects of male aversive pheromones. 7,11-HD also was shown to have a crucial role in heterospecific encounters. Specifically, the species barrier was lost because males of other Drosophila species courted oe(-) D. melanogaster females, and D. simulans males consistently mated with them. Treatment of oe(-) females with 7,11-HD restored the species barrier, showing that a single compound can confer species identity. These results identify a common mechanism for sexual and species recognition regulated by cuticular hydrocarbons.

Nuclear and Mitochondrial Patterns of Introgression between the Parapatric European TreefrogsHyla arboreaandH. intermedia

Postglacial northward expansions from distinct southern European refugia have lead to the formation of several secondary contact zones between species. These zones are providing growing insights into the relevance of introgressive hybridization in shaping patterns of genetic diversity, and into the evolutionary processes involved in the completion of reproductive isolation barriers. Using both nuclear (9 allozyme loci) and mitochondrial (PCR-RFLP of a cytochrome b gene fragment) diagnostic markers we investigated the genetic patterns of variation across a zone of parapatry between the two European treefrogs Hyla arborea and H. intermedia. Neither F1 and F2 hybrids nor backcrosses were identified, indicating the lack of current gene exchange between the two species. However, introgressed alleles were observed in both species and in all markers analysed, which testifies to the occurrence of past events of introgressive hybridization. A wide variation in the frequencies of introgressed alleles was observed b...

Airborne‐pollen pool and mating pattern in a hybrid zone betweenPinus pumilaandP. parvifloravar.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 F(1) hybrid seeds. Both parental species showed a strong assortative mating pattern; F(1) 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.

Low support for separate species within the redpoll complex (Carduelis flammea–hornemanni–cabaret) from analyses of mtDNA and microsatellite markers

The redpoll complex, consisting of three currently recognized species (Carduelis flammea, C. hornemanni and C. cabaret), is polytypic in biometry, morphology, physiology and behaviour. However, previous genetic work has not revealed any indications of genetic differentiation. We analysed sequence variation in the mtDNA control region, and allele frequencies of supposedly faster evolving microsatellites (n=10), in an attempt to detect molecular genetic support for the three species, as well as two subspecies of C. flammea (ssp. flammea and rostrata), within this complex. We used samples from two subspecies of the twite (Carduelis flavirostris, ssp. flavirostris and rufostrigata) as outgroup. We found no structure among redpoll individuals in mtDNA haplotypes or microsatellite allele frequencies, and only marginal differences between redpoll taxa in analyses of molecular variance (AMOVAs) of predefined groups. In contrast, the two twite subspecies constituted two well-supported monophyletic groups. Our study thus strengthens previous indications of low genetic support for current redpoll taxa. Two major alternative interpretations exist. Either redpolls form a single gene pool with geographical polymorphisms possibly explained by Bergmann’s and Gloger’s rules, or there are separate gene pools of recent origin but with too little time elapsed for genetic differentiation to have evolved in the investigated markers. Future studies should therefore examine whether reproductive isolation mechanisms and barriers to gene flow exist in areas with sympatric breeding.

The FERONIA Receptor-like Kinase Mediates Male-Female Interactions During Pollen Tube Reception

In flowering plants, signaling between the male pollen tube and the synergid cells of the female gametophyte is required for fertilization. In the Arabidopsis thaliana mutant feronia (fer), fertilization is impaired; the pollen tube fails to arrest and thus continues to grow inside the female gametophyte. FER encodes a synergid-expressed, plasma membrane-localized receptor-like kinase. We found that the FER protein accumulates asymmetrically in the synergid membrane at the filiform apparatus. Interspecific crosses using pollen from Arabidopsis lyrata and Cardamine flexuosa on A. thaliana stigmas resulted in a fer-like phenotype that correlates with sequence divergence in the extracellular domain of FER. Our findings show that the female control of pollen tube reception is based on a FER-dependent signaling pathway, which may play a role in reproductive isolation barriers.

Differences in karyotype between two sympatric species of Gymnotus (Gymnotiformes: Gymnotidae) from the eastern amazon of Brazil

In this paper we describe the karyotypes of two sympatric species of Gymnotus from Marajó Island, Pará, in the Eastern Amazon basin of Brazil. One of these species, G. mamiraua, has not previously been identified from this area. Karyotype studies demonstrate that G. cf. carapo Linnaeus, has 2n=42 (30M/SM+12ST/A) and G. mamiraua Albert & Crampton, 2n=54 (50M/SM+4ST). The Nucleolar Organizer Region (NOR) was found in the distal region of the short arm of pair 20 in G. cf. carapo and in the interstitial region of pair 1 of G. mamiraua. Both species have centromeric constitutive heterochromatin (CH) in all chromosomes and some in interstitial or distal regions. In both species, 4´6-Diamidino-2- phenylindole (DAPI) staining has shown banding concordant with C-banding, whereas the Chromomycin A3 (CMA3) staining in both species agrees with the NOR location. The new data and review of cytogenetic data in Gymnotus presented here is concordant with a hypothesis that G. carapo is either a complex of populations, or a complex of morphologically similar cryptic species with distinct karyotypes. These taxa may be reproductively isolated by prezygotic reproductive isolation barriers (such as the structure of the Electric Organ Discharge), or by post-zygotic barriers (such as karyotypic formula). The understanding of the cytogenetic structure of G. carapo, combined with associated studies of signal diversity, will yield insight into the mechanisms underlying the origins and maintenance of high species diversity in Gymnotus.

Differences in karyotype between two sympatric species of Gymnotus (Gymnotiformes: Gymnotidae) from the eastern amazon of Brazil

In this paper we describe the karyotypes of two sympatric species of Gymnotus from Marajó Island, Pará, in the Eastern Amazon basin of Brazil. One of these species, G. mamiraua, has not previously been identified from this area. Karyotype studies demonstrate that G. cf. carapo Linnaeus, has 2n=42 (30M/SM+12ST/A) and G. mamiraua Albert & Crampton, 2n=54 (50M/SM+4ST). The Nucleolar Organizer Region (NOR) was found in the distal region of the short arm of pair 20 in G. cf. carapo and in the interstitial region of pair 1 of G. mamiraua. Both species have centromeric constitutive heterochromatin (CH) in all chromosomes and some in interstitial or distal regions. In both species, 4´6-Diamidino-2- phenylindole (DAPI) staining has shown banding concordant with C-banding, whereas the Chromomycin A3 (CMA3) staining in both species agrees with the NOR location. The new data and review of cytogenetic data in Gymnotus presented here is concordant with a hypothesis that G. carapo is either a complex of populations, or a complex of morphologically similar cryptic species with distinct karyotypes. These taxa may be reproductively isolated by prezygotic reproductive isolation barriers (such as the structure of the Electric Organ Discharge), or by post-zygotic barriers (such as karyotypic formula). The understanding of the cytogenetic structure of G. carapo, combined with associated studies of signal diversity, will yield insight into the mechanisms underlying the origins and maintenance of high species diversity in Gymnotus.

Analysis of nucleotide diversity at the cytochrome b and cytochrome oxidase 1 genes at the population, species, and genus levels

Algorithms of nucleotide diversity measures and other measures of genetic divergence at the molecular level are analyzed. Based on a database of p-distances, we have compared genetic divergence of populations (1) and taxa of different rank, such as sibling species (2), species within a genus (3), and species from different genera within a family (4). Based on the theory and algorithms of distance calculation from the primary DNA sequences, as well as the actual distances estimated from literature, it is recommended to use in analysis of experimental data a specific model selected from the eight available ones. The empirical data for more than 24,000 vertebrate and invertebrate species demonstrate that the data series are realistic and interpretable when p-distance or its various estimates are used. This testifies to the applicability of p-distance for most interspecies and intraspecies comparisons of genetic divergence up to the family level by two genes compared. Data on p-distances revealed various and increasing levels of genetic divergence of the sequences of genes Cyt-b and Co-1 in four groups compared. Mean unweighted scores of distances for the four groups were as follows: Cyt-b (1) 1.55 +/- 0.56, (2) 5.52 +/- 1.34, (3) 10.69 +/- 1.34, (4) 18.51 +/- 2.09 and Co-1 (1) 0.55 +/- 0.19, (2) 4.91 +/- 0.83, (3) 9.66 +/- 0.72, (4) 14.69 +/- 1.02. Differences in divergence between the genes themselves at the four levels were also found, although the total mean distances for the two genes did not show statistically significant differences. This conforms to the ample evidence showing different and nonuniform evolution rates of these and other genes and their various regions. The results of the analysis of the nucleotide and allozyme divergence within species and higher taxa of animals, first, are in a good agreement with these results, including data on protein gene markers, and, second, this evidence suggests that in animals, phyletic evolution is likely to prevail at the molecular level, while speciation mainly corresponds to the type D1 geographic model). The prevalence of the D1 speciation mode does not mean that the other modes are absent. There are at least seven various modes of speciation. Recognition of speciation modes is a task that seems to require construction of a quantitative genetic model (theory) of speciation. Although, in view of a vast diversity of the possible causes of reproductive isolating barriers (RIBs) and speciation initiation, as well as the "empirical nature" of the formalized approach, proposed in the present work, some newly arising questions may be left without an answer. Their solution probably lied in increasing the number of descriptors and members of equations, proposed in this study, on the basis of DNA markers and other genomic characteristics.

LEAKY PREZYGOTIC ISOLATION AND POROUS GENOMES: RAPID INTROGRESSION OF MATERNALLY INHERITED DNA

Accurate phylogenies are crucial for understanding evolutionary processes, especially species diversification. It is commonly assumed that "good" species are sufficiently isolated genetically that gene genealogies represent accurate phylogenies. However, it is increasingly clear that good species may continue to exchange genetic material through hybridization (introgression). Many studies of closely related species reveal introgression of some genes without others, often with more rapid introgression of maternally inherited chloroplast or mitochondrial DNA (cpDNA, mtDNA). We seek a general explanation for this biased introgression using simple models of common reproductive isolating barriers (RIBs). We compare empirically informed models of prezygotic isolation (for pre- and postinsemination mechanisms of both female choice and male competition) with postzygotic isolation and demonstrate that rate of introgression depends critically upon type of RIB and mode of genetic inheritance (maternal versus biparental versus paternal). Our frequency-dependent prezygotic RIBs allow much more rapid introgression of biparentally and maternally inherited genes than do commonly modeled postzygotic RIBs (especially maternally inherited DNA). After considering the specific predictions in the context of empirical observations, we conclude that our model of prezygotie RIBs is a general explanation for biased introgression of maternally inherited genomic components. These findings suggest that we should use extreme caution when interpreting single gene genealogies as species phylogenies, especially for cpDNA and mtDNA.

LEAKY PREZYGOTIC ISOLATION AND POROUS GENOMES: RAPID INTROGRESSION OF MATERNALLY INHERITED DNA

Accurate phylogenies are crucial for understanding evolutionary processes, especially species diversification. It is commonly assumed that “good” species are sufficiently isolated genetically that gene genealogies represent accurate phylogenies. However, it is increasingly clear that good species may continue to exchange genetic material through hybridization (introgression). Many studies of closely related species reveal introgression of some genes without others, often with more rapid introgression of maternally inherited chloroplast or mitochondrial DNA (cpDNA, mtDNA). We seek a general explanation for this biased introgression using simple models of common reproductive isolating barriers (RIBs). We compare empirically informed models of prezygotic isolation (for pre- and postinsemination mechanisms of both female choice and male competition) with postzygotic isolation and demonstrate that rate of introgression depends critically upon type of RIB and mode of genetic inheritance (maternal versus biparental versus paternal). Our frequency-dependent prezygotic RIBs allow much more rapid introgression of biparentally and maternally inherited genes than do commonly modeled postzygotic RIBs (especially maternally inherited DNA). After considering the specific predictions in the context of empirical observations, we conclude that our model of prezygotic RIBs is a general explanation for biased introgression of maternally inherited genomic components. These findings suggest that we should use extreme caution when interpreting single gene genealogies as species phylogenies, especially for cpDNA and mtDNA.

Hybridization between the freshwater prawns Macrobrachium rosenbergii (De Man) and M. carcinus (L.)

The production of hybrids between Macrobrachium rosenbergii and other Macrobrachium species has great economical importance for aquaculture, but also has serious environmental implications when M. rosenbergii is present as an exotic species. In this study, intra- and interspecific crosses between M. rosenbergii and M. carcinus were carried out through natural mating and artificial insemination. For the natural crosses, a fertile female was placed inside an aquarium with a mature male and the behavior of animals was monitored. For the artificial insemination assays, male spermatophores were collected by electroejaculation and placed inside the sperm receptacle of a fertile female. The temperature was maintained at 27±2 °C during the study. Firstly, 62 and 45 natural intraspecific crosses were carried out in M. rosenbergii and M. carcinus, with larvae obtained from 32.3% and 57.8% of the crosses, respectively. Thirty-seven natural interspecific reciprocal crosses were also attempted but no mating occurred. A total of 57 and 39 intraspecific crosses were carried out by artificial insemination in M. rosenbergii and M. carcinus, obtaining larva from 22.8% and 41.0% of the crosses, respectively. Finally, 46 interspecific reciprocal crosses were also carried out by artificial insemination. Hybrid zygotes were produced in all the crosses, but embryo development did not progress further than the gastrula stage. This indicates the existence of both behavioral and postzygotic reproductive isolation barriers. These barriers could prevent the hybridization of these species in the natural environment.

Genetic and epigenetic incompatibilities underlie hybrid dysgenesis in Peromyscus.

Crosses between the two North American rodent species Peromyscus polionotus (PO) and Peromyscus maniculatus (BW) yield parent-of-origin effects on both embryonic and placental growth. The two species are approximately the same size, but a female BW crossed with a male PO produces offspring that are smaller than either parent. In the reciprocal cross, the offspring are oversized and typically die before birth. Rare survivors are exclusively female, consistent with Haldane's rule, which states that in instances of hybrid sterility or inviability, the heterogametic sex tends to be more severely affected. To understand these sex- and parent-of-origin-specific patterns of overgrowth, we analysed reciprocal backcrosses. Our studies reveal that hybrid inviability is partially due to a maternally expressed X-linked PO locus and an imprinted paternally expressed autosomal BW locus. In addition, the hybrids display skewing of X-chromosome inactivation in favour of the expression of the BW X chromosome. The most severe overgrowth is accompanied by widespread relaxation of imprinting of mostly paternally expressed genes. Both genetic and epigenetic mechanisms underlie hybrid inviability in Peromyscus and hence have a role in the establishment and maintenance of reproductive isolation barriers in mammals.

Egg size and hybrid syndrome-dependent embryo mortality in Chironomus hybrids (Diptera: Chironomidae)

Female hybrids of the cross Chironomus t. thummi female × Ch. t. piger male which are largely affected by the sterility inducing Rud syndrome were backcrossed with males of both parental strains. The aim of the study was to provide information about those egg volumes that are insufficient for a normal embryogenesis and to ascertain whether in the hybrids the lethally small egg size represents a new abnormal trait of the Rud syndrome. The egg masses obtained contain eggs of very different sizes with volumes ranging from 0.5 nl to 3.49 nl. Embryo mortality is unusually frequent in those eggs of the backcrosses and of the parental strains that have volumes smaller than 1.5 nl. An egg volume of 1.5 nl represents in Ch. thummi the lower limit for those volumes that are sufficient for a normal embryogenesis. Mortality increases with decreasing egg size, reaching 100% in backcross eggs with volumes of 0.99 nl and smaller. Small egg size is a new trait of the Rud syndrome affected thummi female × piger male hybrids. This trait is part of a postzygotic reproductive isolation barrier between thummi and piger and manifests first in the backcrosses. Most backcross eggs show volumes between 1.5 nl and 2.99 nl. Within this volume range the amount of mortality does not depend upon egg volume. Here, embryo death is great in the backcrosses but normal in the parental strains. The high frequency of embryo death in the backcrosses must be predominantly due to the action of the Rud syndrome and a second hybrid syndrome, called HLE syndrome. Since further characteristic traits of these syndromes could be detected in surviving backcross individuals, the study demonstrates the occurrence of the syndromes in this generation also. Therefore, the postzygotic reproductive isolation mechanism of both hybrid syndromes is effective in the hybrids and in their progeny as well.

Bromus pictus of the B. setifolius complex (section Pnigma): numerical taxonomy and chromosome evidence for species rank

Two biotic sympatric taxa of the Bromus setifolius complex, which are presently considered as varieties, were studied using cytological and numerical taxonomy methods. The numerical taxonomy analysis (cluster and principal components analysis) based on 35 characters showed the existence of two phenetically significant groups. One is formed by individuals that correspond to B. setifolius Presl. and the other by individuals with the characters of Bromus pictus Hook. All plants of B. setifolius had 2n = 4x = 28, and all the B. pictus individuals had 2n = 10x = 70. Other cytological differences between the two taxa were found, e.g., karyotype formulae, type of satellites, basic genome length, asymmetry, C-banding patterns, and meiotic chromosome behavior. Cytological differences, which parallel the constant morphological features exhibited by each taxa, justify specific status. Taking into acount other characters, such as perenniality, breeding system, and reproductive isolation barrier, the relationships and evolution of these taxa are discussed. Key words: Bromus setifolius, Bromus pictus, numerical taxonomy, karyotype, C-banding.

Genetic Affinities among the Oncocyclus irises

Crosses were performed between 33 species representing seven recognized trends of divergence in Section Oncocyclus (Siem.) Baker of the genus Iris L. All tested combinations were cross-compatible, and the various F1 interspecific hybrids were fully fertile. We concluded that speciation in this group did not involve the buildup of internal reproductive isolation barriers. The various species of the highly specialized, insect-pollinated Oncocyclus irises are isolated from one another by geographic, ecological, and temporal mechanisms and by preferences of their pollinators.

A study of Avena macrostachya

The morphology, distribution, developmental and reproductive behaviour, cytology, and glumes' epicuticular waxes of Avena macrostachya, a long overlooked species, have been studied and described. In addition, a few interspecific crosses have been attempted but were unsuccessful. Based on unique traits, such as perenniality, cross-fertilization, and quadrivalent-forming autotetraploidy (2n = 4x = 28), it was found to have an isolated position in Avena. It was concluded, however, that at present there is no strong reason to exclude it from the genus. Because of its unique traits, this species may be particularly important in breeding programs when the nature of its reproductive isolation barriers is understood.