Disomic Inheritance Research Articles

High variability and disomic segregation of microsatellites in the octoploid Fragaria virginiana Mill. (Rosaceae).

The objectives of the present study were to develop microsatellite markers for the wild strawberry, Fragaria virginiana, to evaluate segregation patterns of microsatellite alleles in this octoploid species, and assess genetic variability at microsatellite loci in a wild population. A genomic library was screened for microsatellite repeats and several PCR primers were designed and tested. We also tested the use of heterologous primers and found that F. virginiana primers amplified products in cultivated strawberry, Fragaria x ananassa Duch. and Fragaria chiloensis. Similarly, microsatellite loci developed from cultivated strawberry also successfully amplified F. virginiana loci. We investigated four microsatellite loci in detail, three developed from F. virginiana and one from cultivated strawberry. A survey of 100 individuals from a population of F. virginiana in Pennsylvania demonstrated high heterozygosities (H(e) or gene diversity ranged from 0.80 to 0.88 per locus) and allelic diversity (12-17 alleles per locus), but individual plants had no more than two alleles per locus. Segregation patterns in parents and progeny of two controlled crosses at these four loci were consistent with disomic Mendelian inheritance. Together these findings suggest that the genome of F. virginiana is "highly diploidized" and at least a subset of microsatellite loci can be treated as codominant, diploid markers. Significant heterozygote deficiencies were found at three of the four loci for hermaphroditic individuals but for only one locus among females in this gynodioecious species.

Recent duplication of the common carp (Cyprinus carpio L.) genome as revealed by analyses of microsatellite loci.

Genome duplications may have played a role in the early stages of vertebrate evolution, near the time of divergence of the lamprey lineage. Additional genome duplication, specifically in ray-finned fish, may have occurred before the divergence of the teleosts. The common carp (Cyprinus carpio) has been considered tetraploid because of its chromosome number (2n = 100) and its high DNA content. We studied variation using 59 microsatellite primer pairs to better understand the ploidy level of the common carp. Based on the number of PCR amplicons per individual, about 60% of these primer pairs are estimated to amplify duplicates. Segregation patterns in families suggested a partially duplicated genome structure and disomic inheritance. This could suggest that the common carp is tetraploid and that polyploidy occurred by hybridization (allotetraploidy). From sequences of microsatellite flanking regions, we estimated the difference per base between pairs of alleles and between pairs of paralogs. The distribution of differences between paralogs had two distinct modes suggesting one whole-genome duplication and a more recent wave of segmental duplications. The genome duplication was estimated to have occurred about 12 MYA, with the segmental duplications occurring between 2.3 and 6.8 MYA. At 12 MYA, this would be one of the most recent genome duplications among vertebrates. Phylogenetic analysis of several cyprinid species suggests an evolutionary model for this tetraploidization, with a role for polyploidization in speciation and diversification.

Characterization of mixed disomic and polysomic inheritance in the octoploid strawberry (Fragaria x ananassa) using AFLP mapping.

A two-way pseudo-testcross strategy, combined with Single Dose Restriction Fragment (SDRF) marker analysis, was used for genetic mapping in the octoploid cultivated strawberry Fragaria x ananassa (2n = 8 x = 56). Based on a 113 full-sib progeny from a cross between the variety Capitola and the clone CF1116, we generated two parental maps using Amplified Fragment Length Polymorphism (AFLP) markers. Ninety two percent of the markers (727 out of 789) showed ratios corresponding to simplex markers (the majority being SDRF markers), and 8% (62 out of 789) fitted a multiplex ratio. Linkage maps were first established using SDRF markers in coupling phase. The female map comprised 235 markers distributed among 43 co-segregation groups, giving a map size of 1,604 cM. On the male map, 280 markers were assigned to 43 co-segregation groups, yielding a map size of 1,496 cM. Once the co-segregation groups were established, their association was tested using repulsion-phase markers. In total, taking into account associations representing the same linkage groups, 30 linkage groups were detected on the female side and 28 on the male side. On the female map, 68.3% of the pairwise marker linkages were in coupling versus 31.7% in repulsion phase, and the corresponding figures on the male map were 72.2% and 27.8%, respectively. In addition, both groups linked only in the coupling phase and groups linked in the repulsion phase were characterized. The observations suggest that the meiotic behavior of the F. x ananassa genome is neither fully disomic nor fully polysomic, but rather mixed. The genome may not be as completely diploidized as previously assumed.

Linkage mapping in tetraploid willows: segregation of molecular markers and estimation of linkage phases support an allotetraploid structure for Salix alba x Salix fragilis interspecific hybrids.

Salix alba-Salix fragilis complex includes closely related dioecious polyploid species, which are obligate outcrossers. Natural populations of these willows and their hybrids are represented by a mixture of highly heterozygous genotypes sharing a common gene pool. Since nothing is known about their genomic constitution, tetraploidy (2n=4x=76) in willow species makes basic and applied genetic studies difficult. We have used a two-way pseudotestcross strategy and single-dose markers (SDMs) to construct the first linkage maps for both pistillate and staminate willows. A total of 242 amplified fragment length polymorphisms (AFLPs) and 50 selective amplifications of microsatellite polymorphic loci (SAMPL) markers, which showed 1:1 segregation in the F(1) mapping populations, were used in linkage analysis. In S. alba, 73 maternal and 48 paternal SDMs were mapped to 19 and 16 linkage groups covering 708 and 339 cM, respectively. In S. fragilis, 13 maternal and 33 paternal SDMs were mapped in six and 14 linkage groups covering 98 and 321 cM, respectively. For most cosegregation groups, a comparable number of markers linked in coupling and repulsion was identified. This finding suggests that most of chromosomes pair preferentially as occurs in allotetraploid species exhibiting disomic inheritance. The detection of 10 pairs of marker alleles from single parents showing codominant inheritance strengthens this hypothesis. The fact that, of the 1122 marker loci identified in the two male and female parents, the vast majority (77.5%) were polymorphic and as few as 22.5% were shared between parental species highlight that S. alba and S. fragilis genotypes are differentiated. The highly difference between S. alba- and S. fragilis-specific markers found in both parental combinations (on average, 65.3 vs 34.7%, respectively) supports the (phylogenetic) hypothesis that S. fragilis is derived from S. alba-like progenitors.

Formation of 2n gametes in durum wheat haploids: Sexual polyploidization

Allopolyploidy, resulting from interspecific and intergeneric hybridization accompanied by sexual doubling of chromosomes, has played a major role in the evolution of crop plants that sustain humankind. The allopolyploid species, including durum wheat, bread wheat, and oat, have developed a genetic control of chromosome pairing that confers on them meiotic regularity (diploid-like chromosome pairing), and hence reproductive stability, and disomic inheritance. Being natural hybrids, they enjoy the benefits of hybridity as well as polyploidy that make them highly adaptable to diverse environments. Despite the complexities of sexual reproduction, it is widespread among plants and animals. Sexual polyploids are highly successful in nature. Sexual polyploidization is far more efficient than somatic chromosome doubling. Sexual polyploidization effected by functioning of unreduced (2n) gametes in the parental species or in their hybrids has been instrumental in producing our grain, fiber, and oilseed crops. Evidence is presented for the occurrence of sexual polyploidization in durum haploids. ThePh1-induced failure of homoeologous pairing is an important factor in the formation of first division restitution(FDR) nuclei and 2n gametes. The evolutionary and breeding significance of sexual polyploidization is discussed. It is emphasized that three factors, viz.,sexual reproduction, allopolyploidy, and genetic control of chromosome pairing,jointly constitute a perfect recipe for cataclysmic evolution in nature.

Disomic Inheritance, Suppressed Recombination, and Allelic Interactions Govern Apospory in Buffelgrass as Revealed by Genome Mapping

Molecular tools have not identified the gene(s) governing apomixis nor have they been used to successfully transfer the trait to important, sexually reproducing food crops. Several molecular studies addressing apomixis in grasses have used interspecific and intergeneric hybrids. The failure to recover specific P 1 genotypes from these wide crosses can be caused by unfavorable interactions between the gametes, zygote, embryo, endosperm, ana/ or maternal tissue. These interactions can eliminate recombinant genotypes with valuable information towards linkage analyses of the trait. Buffelgrass [Pennisetum ciliare (L.) Link syn. Cenchrus ciliaris L.], a polymorphic species with interfertile apomictic and sexual genotypes, offers an opportunity to genetically map apomixis by means of intraspecific hybrids with euploid genomes. This study reports a linkage map of the apospory region in buffelgrass. Apospory, classified by progeny testing and cytologically observing megagametophytes, mapped to a single locus in the apomictic parent's genome. Two buffelgrass cDNAs (pPAP3A07 and pPAP8C08) and three previously reported apospory markers (UGT197, QH8, and OPC4) were tightly linked (1.4 centimorgans, cM) to the trait. As a tetraploid species (2n = 4x = 36), four copies of each chromosome are expected in buffelgrass. A single homolog and two homeologs were identified for the chromosome carrying apospory, indicating the formation of two bivalents during meiosis and the disomic inheritance of apospory in buffelgrass. Allelic bridges between the parents revealed suppressed recombination in the apospory linkage group. Segregation distortion between a marker on the sexual parent's homolog to the apospory linkage group and a marker on a separate maternal linkage group suggested specific allelic combinations in female gametes affect offspring survival in buffelgrass.

An improved genetic model generates high-resolution mapping of QTL for protein quality in maize endosperm.

The genetic mapping of polymorphic markers in a cross between two inbred plant lines has proven to be a powerful method for detecting quantitative trait loci (QTL) underlying complex traits. However, existing methods of QTL mapping were developed for disomic inheritance of both marker and QTL loci in a diploid population. To map QTL influencing traits expressed in the endosperm, a triploid embryo-nourishing tissue resulting from double fertilization, existing QTL mapping models require modification to consider the trisomic inheritance of the endosperm and the generation difference between the mapping population and the endosperm. Such a model requires simultaneous use of two successive generations, which theoretically can lead to an increase in resolution for QTL mapping compared with the use of a single backcross or F(2) generation. Using a newly developed model based on these considerations, we demonstrate the improved resolution of QTL, influencing protein quality traits in maize endosperm. The increased resolution made possible with this approach makes identified QTL accessible to positional cloning.

Evidence for tetrasomic inheritance in a tetraploid Solanum commersonii (+) S. tuberosum somatic hybrid through the use of molecular markers.

In order to assess the potential for interspecific recombination between the cultivated Solanum tuberosum (tbr) and the sexually isolated wild species Solanum commersonii(cmm), genetic analysis of a F(2) progeny obtained by selfing one tetraploid cmm (+) tbr somatic hybrid was performed through molecular markers. For this purpose, the extent of disomic and/or tetrasomic inheritance of species-specific RAPD and AFLP markers was determined by following their segregation in a 90-genotype progeny, and testing all the possible segregation ratios in a selfed tetraploid progeny. The RAPD analysis performed using 16 primers revealed that the cmm-specific RAPDs were mainly (93.7%) duplex markers and were equally distributed between loci with a disomic (46.7%) and tetrasomic (53.3%) inheritance. The AFLP analysis led to the identification of 272 (58%) informative AFLPs, which were either cmm- or tbr-specific markers. About 63% of cmm-specific AFLPs were duplex loci, most of which (92.6%) were inherited as tetrasomic loci. As regards the tbr-specific AFLPs, the percentage of simplex loci (52.9%) was higher than that of duplex loci (32.6%), and among the latter most (88.5%) were inherited as tetrasomic loci. Overall, 130 duplex markers were found, of which 53.1% were cmm-specific and 46.9% were tbr-specific. Out of 130 markers, 18 (13.8%) were inherited as disomic, and 112 (86.2%) as tetrasomic, loci. This implies that the majority of duplex markers were located on chromosomes which at meiosis tend to randomly pair as bivalents or to form tetravalents. The total number of simplex loci was 119, and most of them (82.3%) were tbr-specific loci. In some cases the observed segregation ratios even allowed us to clearly determine whether a random chromosome or chromatid segregation was detected. This was the case of three cmm-specific RAPDs, 19 cmm- and 25 tbr-specific AFLPs, which fit a 20.8:1 or 2.5:1 ratio, both cases for which a clear random chromatid segregation can be assumed, since they represent the limit of segregation expected when the distance between the locus and the centromere always leads to a cross-over event. The percentage of ascertained crossing-over events was around 37% out of the tetrasomically inherited loci clearly identified (128 loci), a value indicating that the flow of genes from the sexually isolated S. commersonii to the cultivated potato is possible, for at least a large proportion of genes.

Allozyme variation of nonspecific esterases in Russian sturgeon (Acipenser güldenstädti Brandt

Electrophoresis in 7% polyacrylamide gel was used to analyze nonspecific esterases from different organs and tissues of Russian sturgeon. The basic enzymatic activity was observed the four zymogram zones. In sixteen cases the patterns of isozyme distribution in these zones allowed to consider them as allele systems (14 polymorphic and 2 monomorphic) corresponding to the four structural genes. The observed genotype frequencies of the nine of these systems (EST-1* of brain and intestine; EST-2* of serum and intestine; EST-3* of spleen and kidney; and EST-4* of spleen, brain, and heart) were largely concordant with Hardy-Weinberg proportions for codominant disomic inheritance. Genetic control of other polymorphic systems is unknown, but, high enzymatic activity of these esterases, sufficient for qualitative electrophretic detection, permits utilization of these polymorphisms for phenotypic monitoring of Russian sturgeon populations.

Primary Trisomics and SSR Markers as Tools to Associate Chromosomes with Linkage Groups in Soybean

Primary trisomics provide an excellent cytogenetic tool to associate genes and linkage groups with their respective chromosomes. A complete set of 20 primary trisomics (2x + 1 = 41) has been established in soybean [Glycine max (L.) Merr.]. A linkage map of soybean with 20 consensus linkage groups has recently been defined. Because simple sequence repeat (SSR) markers map to defined single positions in the soybean genome, the association of a SSR locus with a chromosome will provide an unambiguous association of a linkage group to a specific chromosome. The objective of this work was to demonstrate the use of SSR markers to associate linkage groups with chromosomes by means of primary trisomics. One population of F2 plants was developed from an F1 hybrid trisomic for chromosome 13 (Triplo 13) and a second F2 population was obtained from a F1 hybrid trisomic for chromosome 5 (Triplo 5). Polymorphic SSR markers from different consensus linkage groups were tested on a subset of 20 plants from each population to identify markers that appeared to show segregation that deviated from normal (1:2:1) disomic inheritance. Markers not associated with the specific chromosome segregated in a disomic (1:2:1) fashion. Markers identified in this manner were further examined in the complete population of F2 plants to identify those that demonstrated trisomic segregation (6:11:1). By this approach, Triplo 13 was associated with linkage group F and Triplo 5 with linkage group A1. This result was verified by the examination of seven SSR loci on linkage group F and eight loci from linkage group A1 with each showing trisomic segregation with the Triplo 13‐ and Triplo 5‐derived F2 populations, respectively. These results demonstrate the first association of molecular linkage groups with chromosomes in soybean and indicate that SSR markers provide a tool to associate the remaining 18 trisomics with their respective linkage groups.

Detecting and mapping repulsion-phase linkage in polyploids with polysomic inheritance

It has been suggested that ratios of coupling- to repulsion-phase linked markers can be used to distinguish between allopolyploids and autopolyploids, because repulsion-phase linkages are much more difficult to detect in autopolyploids with polysomic inheritance than allopolyploids with disomic inheritance. In this report, we analyze the segregation pattern of repulsion-phase linked markers in polyploids without complete preferential pairing. The observed repulsion-phase recombination fraction (R) in such polyploids is composed of a fraction due to crossing-over (Rc) and another fraction due to independent assortment (Ri). Ri is the minimum distance that can be detected between repulsion-phase linked markers. Because Ri is high in autopolyploids (0.3373, 0.4000, 0.4286 and 0.4444) for autopolyploids of 2n=4x, 6x, 8x and 10x), large population sizes are required to reliably detect repulsion linkages. In addition, the default linkage used in mapping-programs must be greater than the corresponding Ri to determine whether a polyploid is a true autopolyploid. Unfortunately, much lower default linkages than the Ris have been used in recent polyploid studies to determine polyploid type, and markers have been incorporated into polyploid maps based on the R values. Herein, we describe how mapping repulsion linkages can result in spurious results, and present methods to accurately detect the degree of preferential pairing in polyploids using repulsion linkage analysis.

Inheritance of tristyly in Oxalis tuberosa (Oxalidaceae)

Frequencies of floral morphs in progenies obtained from a complete set of diallelic crosses among three accessions of tristylous, octoploid oca (Oxalis tuberosa) were used for a Mendelian analysis of floral morph inheritance. The frequencies observed had the best fit to a model of tetrasomic inheritance with two diallelic factors, S, s and M, m, with S being epistatic over M. No explanation could be found for the unexpected formation of a small percentage of short-styled individuals in crosses between the mid-styled and the long-styled parent. For the acceptance of models of disomic and octosomic inheritance several additional assumptions would have to be made and therefore these modes of inheritance are less likely. Dosage-dependent inheritance of floral morph was rejected. Only a small frequency (36%) of the cross progenies flowered, in contrast to the greater propensity for flowering of O. tuberosa accessions held at gene banks.

RFLP marker analysis supports tetrasonic inheritance in Lotus corniculatus L.

Lotus corniculatus is a tetraploid (2n=4x=24) perennial forage legume and has been reported to have tetrasomic inheritance for several traits, although it has also been reported to show disomic inheritance. Molecular markers were used to clarify whether tetrasomic inheritance, disomic inheritance, or a combination of both, was found within an F2 population arising from a cross between two diverse L. corniculatus accessions. The inheritance of ”tetra-allelic” RFLP markers (markers with four segregating bands) indicated that disomic inheritance could not account for the phenotypic F2 classes observed, and that only tetrasomic inheritance would explain the observed results. Goodness of fit tests for ”tetra-allelic” and ”tri-allelic” (three segregating bands) RFLP marker data suggested support for chromosomal-type tetrasomic inheritance. RFLP genotypes interpreted from autoradiographic signal intensity provided additional support for tetrasomic inheritance and the occurrence of preferential pairing between parental chromosomes. Bivalent pairing was predominant in the two parental lines and their F1 hybrid in cytological analyses. L. corniculatus has been classified as both an autotetraploid and an allotetraploid species. RFLP evidence of tetrasomic inheritance gives support for L. corniculatus being classified as an autotetraploid species. Even though bivalent pairing occurs, as seen in other autotetraploid species, pairing between any of the four homologous chromosomes is possible. Preferential pairing in the F1 hybrid suggests that genome differentiation appears to be minimal between homologs within an accession, while genome differentiation is greater between homologs from different accessions of this genetically diverse species.

Inheritance of microsatellite loci in the polyploid lake sturgeon (Acipenser fulvescens).

Inheritance in the expression of amplicons for four microsatellite primer pairs was determined using 10 families created from gametes of wild lake sturgeon (Acipenser fulvescens). Loci Afu34 and Afu68 expressed a maximum of two even-intensity bands per individual and had progeny genotype ratios that fit disomic inheritance (P > 0.05). Some variation exhibited at Afu34 and Afu68 was attributable to a null allele. Genotype expression at both loci also indicated that one female parent had transmitted unreduced gametes. Primer Afu39 amplified products that exhibited four gene doses, where genotype counts fit expected ratios for disomic inheritance (P > 0.05) indicating amplification of products from two disomic loci that share alleles. Meiotic drive was evident at the Afu39 loci based on a test for random segregation (P < 0.05). Only the expression of Afu19 gave evidence of tetrasomic inheritance based on a single progeny potentially produced by a double reduction gamete. No evidence for proposed octoploid inheritance was observed.

Inheritance of resistance in smooth bromegrass to the crown rust fungus.

Common smooth bromegrass (Bromus inermis Leyss.) is octoploid, 2n = 8x = 56, with a genome structure of AAAAB1B1B2B2. Tetrasomic inheritance patterns have been observed in smooth bromegrass, but disomic inheritance is also expected from cytologic observations. Smooth bromegrass is susceptible to the crown rust fungus (Puccinia coronata Corda.). The objective of this study was to determine the inheritance of smooth bromegrass resistance to P. coronata. Seven smooth bromegrass clones, three susceptible and four resistant, were selfed and crossed in a diallel with bulked reciprocals. Inoculations were made with a population of P. coronata from PL-BDR1 smooth bromegrass. Resistance of smooth bromegrass to this population of P. coronata is complex. At least three genes appear to be involved in this host-pathogen interaction, one tetrasomic dominant gene which determines susceptibility (S) and two dominant genes (R1 and R2) that may be complementary and could be inherited either tetrasomically or disomically. Other genes may be involved in the smooth bromegrass-P. coronata interaction, possibly accounting for the lack of fit to expected ratios of some progeny. Heterogeneity for avirulence phenotype in the pathogen population may also have contributed to lack of fit of some progeny. Multiple resistance genes were detected because a pathogen population, likely consisting of genotypes with different genes for virulence, was used to challenge the host.

The Cytonuclear Effects of Facultative Apomixis: II. Definitions and Dynamics of Disequilibria in Tetraploid Populations

We develop a cytonuclear framework for tetraploid populations in which a diallelic nuclear marker exhibits tetrasomic inheritance. This system requires two separate parameterizations, with six cytonuclear disequilibria (nonrandom associations) in tetraploid individuals and four in their diploid gametes. Double reduction during meiosis adds further complexity by causing gametic output to vary with the distance of the nuclear locus from the centromere. We derive and analyze dynamical solutions for the disequilibria under generalized mixed mating, with any combination of apomixis, selfing, and outcrossing, with and without double reduction. As in comparable diploid systems, all disequilibria ultimately decay to zero, unless nuclear and cytoplasmic alleles are nonrandomly associated and outcrossing is absent, in which case permanent associations result. Selfing and apomixis retard the decay of disequilibria (or approach to equilibrium), and often to the same extent. In contrast, double reduction can accelerate the loss of tetraploid cytonuclear associations, but only negligibly in hybrid zones, and this loss is never faster than in diploids. Only in the absence of allelic associations or outcrossing is the asymptotic approach to equilibrium differentially affected by apomixis and selfing or slower under tetrasomic than disomic inheritance. To facilitate empirical applications, we also examine tetraploid hybrid zone dynamics and offer practical guidelines for experimental design and data analysis, showing how the consequences of the mating system alone provide a valuable baseline for drawing evolutionary inferences from the observed patterns of cytonuclear associations.

AmphidiploidBrassica junceacontains conserved progenitor genomes

To perform a detailed study of genome evolution in the natural Brassica amphidiploid B. juncea, we have constructed two linkage maps based on RFLP (restriction fragment length polymorphism) markers; one generated from a cross between a resynthesized B. juncea (a chromosome doubled interspecific B. rapa x B. nigra hybrid) and a natural B. juncea cultivar, the other from a cross between two B. juncea cultivars. By using a common cultivar in both crosses, the two maps could be unambiguously integrated. All loci exhibited disomic inheritance of parental alleles in the natural x resynthesized cross, showing that B. rapa chromosomes paired exclusively with their A-genome homologues in B. juncea and that B. nigra chromosomes likewise paired with their B-genome homologues. The maps derived from the two crosses were also perfectly collinear. Furthermore, these maps were collinear with maps of the diploid progenitor species (B. nigra and B. rapa) produced using the same set of RFLP probes. These data indicate that the genome of B. juncea has remained essentially unchanged since polyploid formation. Our observations appear to refute the suggestion that the formation of polyploid genomes is accompanied by rapid change in genome structure.

A saturated genetic linkage map of rubber tree (Hevea spp.) based on RFLP, AFLP, microsatellite, and isozyme markers

The first genetic map for Hevea spp. (2n=36) is presented here. It is based on a F1 progeny of 106 individuals allowing the construction of a female, a male, and a synthetic map according to the pseudo-testcross strategy. Progeny were derived from an interspecific cross between PB260, a H. brasiliensis cultivated clone, and RO38, a H. brasiliensis×H. benthamiana interspecific hybrid clone. The disomic inheritance observed for all the codominant markers scattered on the 2n=36 chromosomes revealed that Hevea behaves as diploids. Homologous linkage groups between the two parental maps were merged using bridge loci. A total of 717 loci constituted the synthetic map, including 301 RFLPs, 388 AFLPs, 18 microsatellites, and 10 isozymes. The markers were assembled into 18 linkage groups, thus reflecting the basic chromosome number, and covered a total distance of 2144 cM. Nine markers were found to be unlinked. Segregation distortion was rare (1.4%). Average marker density was 1 per 3 cM. Comparison of the distance between loci in the parental maps revealed significantly less meiotic recombination in the interspecific hybrid male parent than in the female parent. Hevea origin and genome organisation are discussed.

Single-locus inheritance in the allotetraploid Coffea arabica L. and interspecific hybrid C. arabica x C. canephora.

Molecular cytogenetic analysis has indicated that Coffea arabica is an amphidiploid formed from the hybridization between two closely related diploid progenitor species, C. canephora and C. eugenioides. Our aim was to determine the mode of inheritance in C. arabica and in a tetraploid interspecific hybrid (called arabusta) between C. arabica and C. canephora as revealed by segregation analyses of restriction fragment length polymorphism (RFLP) loci markers. The observed RFLP allele segregations in an F(2) progeny of C. arabica conform to disomic inheritance as expected, with regular bivalent pairing of homologous chromosomes in the F1 hybrid. In contrast, RFLP loci followed tetrasomic inheritance in the arabusta interspecific hybrid, although bivalents have been reported to predominate greatly at meiosis in its hybrid. These results suggest that homologous chromosomes do not pair in C. arabica, not as a consequence of structural differentiation, but because of the functioning of pairing regulating factors. Moreover, the arabusta hybrid seems to offer the possibility of gene exchange between the homologous genomes.

The allotetraploid invasive weedBromus hordeaceus L. (Poaceae): Genetic diversity, origin and molecular evolution

Bromus hordeaceus (sectionBromus, Poaceae), a predominantly self-fertilizing tetraploid (2n=28), is an annual weed native to the Mediterranean Basin, which now has a world-wide distribution. High morphological variation led to the recognition of four subspecies, three of which correlated with habitat-type. We examined genetic diversity at enzyme loci in 15 populations from the Mediterranean and the Atlantic region. Although sampled over a larger range of ecological and geographical conditions, the North-African populations appeared less genetically differentiated than populations from Brittany, suggesting higher levels of gene flow among the first ones (Nm=3.756 and 1.066 respectively). No genetic differentiation was encountered among the four subspecies. The populations were homozygous at homologous loci, suggesting high rates of selfing, but they frequently exhibited fixed intergenomic heterozygosity. The meiotic chromosome behaviour and disomic inheritance encountered are in accordance with the previously proposed allopolyploid origin of the species. The diploidsB. arvensis andB. scoparius have been previously implicated in the parentage ofB. hordeaceus on the basis of morphology and serology. We comparedB. hordeaceus with related diploid species belonging to the same section (sectionBromus) using different sources of data (flow cytometry, karyotypes, RAPD and DNA sequences). Molecular phylogeny based on internal transcribed spacer sequences of nuclear ribosomal genes provided the first clear scheme of relationships among monogenomic species of the section. A new hypothesis is proposed concerning the origin ofB. hordeaceus: We found that it diverged earlier than all other species of sectionBromus excluding the diploidB. caroli-henrici which is basal in this group. The 13 autapomorphies accumulated byB. hordeaceus, and the absence of intra-individual sequence heterogeneity are also consistent with the relatively ancient origin of the species within the section.