Cytogenetic events in the endosperm of amphiploid Avena magna × A. longiglumis

This study analysed cytogenetic events occurring in the syncytial endosperm of the Avena magna H. C. Murphy & Terrell × Avena longiglumis Durieu amphiploid, which is a product of two wild species having different genomes. Selection through the elimination of chromosomes and their fragments, including those translocated, decreased the level of ploidy in the endosperm below the expected 3n, leading to the modal number close to 2n. During intergenomic translocations, fragments of the heterochromatin-rich C-genome were transferred to the D and Al genomes. Terminal and non-reciprocal exchanges dominated, whereas other types of translocations, including microexchanges, were less common. Using two probes and by counterstaining with DAPI, the A. longiglumis and the rare exchanges between the D and Al genomes were detected by GISH. The large discontinuity in the probe labelling in the C chromosomes demonstrated inequality in the distribution of repetitive sequences along the chromosome and probable intragenomic rearrangements. In the nucleus, the spatial arrangement of genomes was non-random and showed a sectorial-concentric pattern, which can vary during the cell cycle, especially in the less stable tissue like the hybrid endosperm.

Cytogenetic events in the endosperm of amphiploidAvena maroccana×A. longiglumis

This study analysed cytogenetic events occurring in the syncytial endosperm of theAvena maroccanaGand. ×Avena longiglumisDur. amphiploid, which is a product of two wild species having different genomes. Selection through the elimination of chromosomes and their fragments, including those translocated, decreased the level of ploidy in the endosperm below the expected 3n, leading to the modal number close to 2n. During intergenomic translocations, fragments of the heterochromatin-rich C-genome were transferred to the A and Al genomes. Terminal and non-reciprocal exchanges dominated, whereas other types of translocations, including microexchanges, were less common. Using two probes and by counterstaining with DAPI, theA. longiglumisand the rare exchanges between the A and Al genomes were detected by GISH. The large discontinuity in the probe labelling in the C chromosomes demonstrated inequality in the distribution of repetitive sequences along the chromosome and probable intragenomic rearrangements. In the nucleus, the spatial arrangement of genomes was non-random and showed a sectorial-concentric pattern, which can vary during the cell cycle, especially in the less stable tissue like the hybrid endosperm.

New evidence confirming the CD genomic constitutions of the tetraploid Avena species in the section Pachycarpa Baum

The tetraploid Avena species in the section Pachycarpa Baum, including A. insularis, A. maroccana, and A. murphyi, are thought to be involved in the evolution of hexaploid oats; however, their genome designations are still being debated. Repetitive DNA sequences play an important role in genome structuring and evolution, so understanding the chromosomal organization and distribution of these sequences in Avena species could provide valuable information concerning genome evolution in this genus. In this study, the chromosomal organizations and distributions of six repetitive DNA sequences (including three SSR motifs (TTC, AAC, CAG), one 5S rRNA gene fragment, and two oat A and C genome specific repeats) were investigated using non-denaturing fluorescence in situ hybridization (ND-FISH) in the three tetraploid species mentioned above and in two hexaploid oat species. Preferential distribution of the SSRs in centromeric regions was seen in the A and D genomes, whereas few signals were detected in the C genomes. Some intergenomic translocations were observed in the tetraploids; such translocations were also detected between the C and D genomes in the hexaploids. These results provide robust evidence for the presence of the D genome in all three tetraploids, strongly suggesting that the genomic constitution of these species is DC and not AC, as had been thought previously.

New evidence concerning the genome designations of the AC(DC) tetraploid Avena species

The tetraploid Avena species in the section Pachycarpa Baum, including A. insularis, A. maroccana, and A. murphyi, are thought to be involved in the evolution of hexaploid oats; however, their genome designations are still being debated. Repetitive DNA sequences play an important role in genome structuring and evolution, so understanding the chromosomal organization and distribution of these sequences in Avena species could provide valuable information concerning genome evolution in this genus. In this study, the chromosomal organizations and distributions of six repetitive DNA sequences (including three SSR motifs (TTC, AAC, CAG), one 5S rRNA gene fragment, and two oat A and C genome specific repeats) were investigated using non-denaturing fluorescence in situ hybridization (ND-FISH) in the three tetraploid species mentioned above and in two hexaploid oat species. Preferential distribution of the SSRs in centromeric regions was seen in the A and D genomes, whereas few signals were detected in the C genomes. Some intergenomic translocations were observed in the tetraploids; such translocations were also detected between the C and D genomes in the hexaploids. These results provide robust evidence for the presence of the D genome in all three tetraploids, strongly suggesting that the genomic constitution of these species is DC and not AC, as had been thought previously.

Genomic constitution and intergenomic translocations in the Elymus dahuricus complex revealed by multicolor GISH

Multicolor genomic in situ hybridization was used to investigate the genomic constitution and intergenomic translocations in the Elymus dahuricus complex. The genomic constitution of species of the E. dahuricus complex was confirmed as StYH. H/Y and H/St intergenomic translocations were identified in the present study, with 7H and 1Y chromosomes involved in reciprocal translocations for all the accessions investigated in the complex. We propose that the translocations in the E. dahuricus complex are species-specific, associated with allopolyploidy, and may serve as important structural alterations for allopolyploid stability. Furthermore, they may help to restore fertility and nucleocytoplasmic compatibility in a newly formed polyploid and facilitate the successful establishment of E. dahuricus as a stable species. It was found that more chromosomes were involved in translocations and more types of intergenomic translocations were observed in the high altitude (4150 m) population Y 2228 than in populations from relatively lower altitudes (2600–3800 m). We speculate that more complicated genomic changes were associated with escalating altitudes in the Tibetan Plateau. These genomic changes contribute to promote the genetic variability and enable the newly formed allopolyploids to adapt to more changeable and harsher environments during the evolution of a polyploid species, thus facilitating their rapid and successful establishment in nature.

Visualization of U and M genome chromosomes by multicolour genomic in situ hybridization in Aegilops biuncialis and Triticum aestivum-Ae. biuncialis amphiploids

The multicolour genomic in situ hybridization (mcGISH) method was improved in order to visualize the U b and M b genomes of Aegilops biuncialis Vis. (2n=4x=28, U b U b M b M b ). Hybridization probes prepared from the diploid U and M genome donors, Ae. umbellulata and Ae. comosa , resulted in clear hybridization signals on the U and M chromosomes in Ae. biuncialis . The random primed labelling method made it possible to decrease the blocking ratio to 1:30. McGISH allowed the simultaneous discrimination of individual Ae. biuncialis genomes and wheat chromosomes in γ-irradiated Triticum aestivum-Ae. biuncialis amphiploids (2n=70; AABBDDU b U b M b M b ). Dicentric chromosomes, terminal and interstitial translocations and centric fusions were detected in the irradiated generation. The irradiation-induced wheat- Ae. biuncialis intergenomic translocations will facilitate the successful introgression of useful agronomic traits into bread wheat.

Detection of intergenomic chromosome rearrangements in irradiated Triticum aestivum – Aegilops biuncialis amphiploids by multicolour genomic in situ hybridization

The frequency and pattern of irradiation-induced intergenomic chromosome rearrangements were analysed in the mutagenized (M0) and the first selfed (M1) generations of Triticum aestivum L. – Aegilops biuncialis Vis. amphiploids (2n = 70, AABBDDUbUbMbMb) by multicolour genomic in situ hybridization (mcGISH). mcGISH allowed the simultaneous discrimination of individual Ae. biuncialis genomes and wheat chromosomes. Dicentric chromosomes, fragments, and terminal translocations were most frequently induced by γ-irradiation, but centric fusions and internal exchanges were also more abundant in the treated plants than in control amphiploids. Rearrangements involving the Ub genome (Ub-type aberrations) were more frequent than those involving the Mb genome (Mb-type aberrations). This irradiation sensitivity of the Ub chromosomes was attributed to their centromeric or near-centromeric regions, since Ub-type centric fusions were significantly more abundant than Mb-type centric fusions at all irradiation doses. Dicentrics completely disappeared, but centric fusions and translocations were well transmitted from M0 to M1. Identification of specific chromosomes involved in some rearrangements was attempted by sequential fluorescence in situ hybridization with a mix of repeated DNA probes and GISH on the same slide. The irradiated amphiploids formed fewer seeds than untreated plants, but normal levels of fertility were recovered in their offspring. The irradiation-induced wheat – Ae. biuncialis intergenomic translocations will facilitate the successful introgression of drought tolerance and other alien traits into bread wheat.

Transmission of maize chromosome 9 rearrangements in oat–maize radiation hybrids

Oat–maize radiation hybrids are oat (Avena sativa L.) plants carrying radiation-induced subchromosome fragments of a given maize (Zea mays L.) chromosome. Since first-generation radiation hybrids contain various maize chromosome rearrangements in a hemizygous condition, variation might be expected in the transmission of these rearrangements to subsequent generations. The transmission and integrity of maize chromosome 9 rearrangements were evaluated in progenies of 30 oat–maize radiation hybrids by using a series of DNA-based markers and by genomic in situ hybridization. Maize chromosome 9 rearrangements were reisolated by self-fertilization in 24 of the 30 radiation hybrid lineages. Normal and deleted versions of maize chromosome 9 were transmitted at similar frequencies of 9.1% and 7.6%, respectively, while intergenomic translocations were transmitted at a significantly higher frequency of 47.6%. Most lines (93%) that inherited a rearrangement had it in the hemizygous condition. Lines with a rearrangement in the homozygous state (7%) were only identified in lineages with intergenomic translocations. Homozygous lines are more desirable from the perspective of stock maintenance, since they may stably transmit a given rearrangement to a subsequent generation. However, their isolation is not strictly required, since hemizygous lines can also be used for genome mapping studies.Key words: Avena sativa, Zea mays, addition lines, chromosome rearrangements.