Synthetic Allopolyploids Research Articles

Incorrect recombination partner associations contribute to meiotic instability of neo-allopolyploid Arabidopsis suecica.

Combining two or more related homoeologous genomes in a single nucleus, newly formed allopolyploids must rapidly adapt meiosis to restore balanced chromosome segregation, production of euploid gametes and fertility. The poor fertility of such neo-allopolyploids thus strongly selects for the limitation or avoidance of genetic crossover formation between homoeologous chromosomes. In this study, we have reproduced the interspecific hybridization between Arabidopsis thaliana and Arabidopsis arenosa leading to the allotetraploid Arabidopsis suecica and have characterized the first allopolyploid meioses. First-generation neo-allopolyploid siblings vary considerably in fertility, meiotic behavior and levels of homoeologous recombination. We show that centromere dynamics at early meiosis is altered in synthetic neo-allopolyploids compared with evolved A. suecica, with a significant increase in homoeologous centromere interactions at zygotene. At metaphase I, the presence of multivalents involving homoeologous chromosomes confirms that homoeologous recombination occurs in the first-generation synthetic allopolyploid plants and this is associated with a significant reduction in homologous recombination, compared to evolved A. suecica. Together, these data strongly suggest that the fidelity of recombination partner choice, likely during the DNA invasion step, is strongly impaired during the first meiosis of neo-allopolyploids and requires subsequent adaptation.

Homeologue differential expression in the flavonoid biosynthetic pathway underlies flower colour variation in natural and synthetic polyploids of Nicotiana tabacum (Solanaceae)

Abstract Homeologue expression bias occurs when one progenitor copy of a gene is expressed at a higher level than the other in allopolyploids. Morphological variation, including differences in flower colour, exists between natural and synthetic allopolyploids of Nicotiana tabacum and their progenitors. In this study, we use a comparative transcriptomic approach to investigate gene expression differences as well as homeologue bias in the flavonoid biosynthetic pathway (FBP) in these accessions. We do not observe reciprocal homeologue bias between dark and light pink allopolyploids, but the production of light pink flowers is correlated with high FLAVONOL SYNTHASE:DIHYDROFLAVONOL-4-REDUCTASE (FLS:DFR) ratio at 60% of anthesis length due to delayed activation of DFR in these accessions. We do find that natural allopolyploids have stronger homeologue bias than synthetic allopolyploids in both FBP genes and across the transcriptome. While there is no overall subgenome dominance, there is a bias towards expression of N. tomentosiformis homeologues in FBP genes; however, the magnitude of this bias is reduced in allopolyploids compared to the progenitors, suggesting that N. sylvestris homeologues play an active role in the development of flower colour in N. tabacum allopolyploids. In addition, synthetic allopolyploids tend to exhibit trans regulation of homeologues whereas natural allopolyploids often have evolved cis-regulatory differences between homeologues since their origin.

Genome Shock in a Synthetic Allotetraploid Wheat Invokes Subgenome-Partitioned Gene Regulation, Meiotic Instability and Karyotype Variation.

It becomes increasingly evident that interspecific hybridization at homoploid level or coupled with whole genome duplication (i.e., allopolyploidization) has played a major role in biological evolution. Yet, the direct impact of hybridization and allopolyploidization on genome structure and function, phenotype, and fitness remains to be fully understood. Synthetic hybrids and allopolyploids are trackable experimental systems to address this issue. Here, we resynthesized a pair of reciprocal F1 hybrids and corresponding reciprocal allotetraploids using the two diploid progenitor species, Triticum urartu (AA) and Aegilops tauschii (DD), of bread wheat (Triticum aestivum L., BBAADD). By comparing phenotypes related to growth, development and fitness, andanalyzing genome expression in both hybrids and allotetraploids in relation to parents, we find the types and trends of karyotype variation in the immediately formed allotetraploids, are correlated with both meiosis instability and chromosome- and subgenome-biased expression. We document clear advantages of allotetraploids over diploid F1 hybrids in several morphological traits including fitness, which mirror the tissue- and developmental stage-dependent subgenome-partitioning of the allotetraploids. The allotetraploids are meiotically unstable primarily due to homoeologous pairing that varies dramatically among the chromosomes. Nonetheless, the manifestation of organismal karyotype variation and occurrence of meiotic irregularity are not concordant, suggesting a role of functional constraints probably imposed by subgenome- and chromosome-biased gene expression. Our results provide new insights into the direct impacts and consequences of hybridization and allopolyploidization, which are relevant to evolution and likely informative to crop improvement by synthetic polyploid approaches.

Phenotypic trait variation in the North American Tragopogon allopolyploid complex.

Recently formed allopolyploids Tragopogon mirus and T. miscellus and their diploid parental species, T. dubius, T. porrifolius, and T. pratensis, offer a rare opportunity to study the earliest stages of allopolyploidy. The allopolyploid species have also been resynthesized, allowing comparisons between the youngest possible allopolyploid lineages and their natural, established counterparts. For the first time we compare phenotypic traits on a large scale in Tragopogon diploids, natural allopolyploids, and three generations of synthetic allopolyploids. Our large common-garden experiment measured traits in growth, development, physiology, and reproductive fitness and analyzed differences between allopolyploids and their parental species and between synthetic and natural allopolyploids. As in many polyploids, the allopolyploid species had some larger physical traits and a higher capacity for photosynthesis than diploid species. Reproductive fitness traits were variable and inconsistent. Allopolyploids had intermediate phenotypes compared to their diploid parents in several traits, but patterns of variation often varied between allopolyploid complexes. Resynthesized and natural allopolyploid lines generally showed minor to non-existent trait differences. In Tragopogon, allopolyploidy results in some typical phenotypic changes, including gigas effects and increased photosynthetic capacity. Being polyploid did not produce a significant reproductive advantage. Comparisons between natural and synthetic T. mirus and T. miscellus are consistent with very limited, idiosyncratic phenotypic evolution following allopolyploidization. This article is protected by copyright. All rights reserved.

Identification of a novel Carassius amphitetraploid clone and its transcriptomic responses to herpesvirus infection

A major threat to gibel carp aquaculture is an epizootic disease caused by C. auratus herpesvirus (CaHV). Therefore, breeding of disease resistant strain is one of significant needs for the industry. Polyploids often show enhanced tolerance to abiotic or biotic stress. However, the effects of WGD are complex and the underlying mechanisms remain elusive. In this study, we first synthesized a group of novel amphitetraploids via crossing the eggs of gynogenetic C. gibelio clone A+ with the trypsin-treated sperm of sexual C. cuvieri and then established three gynogenetic amphitetraploid clones with different transferrin phenotypes. They all showed intermediate resistance to CaHV in relative to their parents. Subsequently, we uncovered distinct transcriptomic response to CaHV infection in the one amphitetraploid clone (CaA4n-I) compared with their parents. Interestingly, the coexpression gene network analysis showed that amphitetraploid CaA4n-I and their paternal C. cuvieri adopted different anti-viral strategies via up-regulating the expression of positive factors or down-regulating the expression of negative factors in immune response respectively. Moreover, we revealed the complicated changes of expression patterns, including additivity, maternal expression level dominance, paternal expression level dominance, transgressive up-regulation, and transgressive down-regulation, in the amphitetraploid CaA4n-I. After CaHV infection, the proportions of maternal expression level dominance and transgressive up-regulation increased and more C. cuvieri-derived homeologous genes altered expression patterns in the amphitetraploid CaA4n-I. The current study not only produces a candidate strain for C. auratus aquaculture, but also provides a transcriptomic picture how a newly synthetic animal allopolyploid to response to virus infection.

Creating and De Novo Improvement of New Allopolyploid Crops for Future Agriculture

The development of climate change resilient crops is conducive to meeting the increasing threat of supporting the growing world population. Polyploidy occupies an important position in angiosperm evolution, as a key factor that shapes plant biodiversity, growth vigor, environmental adaptation, and emerging chemical compounds. In this review, we outlined the development and application of creating new allopolyploids using sexual and asexual approaches and their potential benefits and problems. We described how polyploidization caused strict genome modification at cytogenetic, genetic, and epigenetic levels with emphasis on the latest update on genome assembly of newly synthesized allopolyploids. Despite the success in creating new allopolyploids in many genera, it occasionally gave rise to undesirable traits to impact the utilization of newly synthetic allopolyploids. Recent developments in the de novo domestication of wild species through genome editing provide a route to create new crops to secure the global food supply. Following the strategy, de novo improvement of newly synthetic allopolyploids using genome editing could be galvanized to rapidly improve newly synthesized allopolyploids to meet agriculture demands and enable plant breeders to keep pace with global changes.

Characterization and acceleration of genome shuffling and ploidy reduction in synthetic allopolyploids by genome sequencing and editing

Polyploidy and the subsequent ploidy reduction and genome shuffling are the major driving forces of genome evolution. Here, we revealed short-term allopolyploid genome evolution by sequencing a synthetic intergeneric hybrid (Raphanobrassica, RRCC). In this allotetraploid, the genome deletion was quick, while rearrangement was slow. The core and high-frequency genes tended to be retained while the specific and low-frequency genes tended to be deleted in the hybrid. The large-fragment deletions were enriched in the heterochromatin region and probably derived from chromosome breaks. The intergeneric translocations were primarily of shortfragments dependent on homoeology, indicating a gene conversion origin. To accelerate genome shuffling, we developed an efficient genome editing platform for Raphanobrassica. By editing Fanconi Anemia Complementation Group M (FANCM) genes, homoeologous recombination, chromosome deletion and secondary meiosis with additional ploidy reduction were accelerated. FANCM was shown to be a checkpoint of meiosis and controller of ploidy stability. By simultaneously editing FLIP genes, gene conversion was precisely introduced, and mosaic genes were produced around the target site. This intergeneric hybrid and genome editing platform not only provides models that facilitate experimental evolution research by speeding up genome shuffling and conversion but also accelerates plant breeding by enhancing intergeneric genetic exchange and creating new genes.

Rapid and visual monitoring of alien sequences using crop wild relatives specific oligo-painting: The case of cucumber chromosome engineering

Wild species related to domesticated crops (crop wild relatives, or CWRs) represent a high level of genetic diversity that provides a practical gene pool for crop pre-breeding employed to address climate change and food demand challenges globally. Nevertheless, rapid identifying and visual tracking of alien chromosomes and sequences derived from CWRs have been a technical challenge for crop chromosome engineering. Here, a species-specific oligonucleotide (oligo) pool was developed by using the reference genome of Cucumis hystrix (HH, 2n = 2x = 24), a wild species carrying many favorable traits and interspecific compatibility with cultivated cucumber (C. sativus, CC, 2n = 2x = 14). These synthetic double-stranded oligo probes were applied to validate the assembly and characterize the chromosome architectures of C. hystrix, as well as to rapidly identify C. hystrix-chromosomes in diverse C. sativus-hystrix chromosome-engineered germplasms, including interspecific hybrid F1 (HC), synthetic allopolyploids (HHCC, CHC, and HCH) and alien additional lines (CC-H). Moreover, a ∼2Mb of C. hystrix-specific sequences, introduced into cultivated cucumber, were visualized by CWR-specific oligo-painting. These results demonstrate that the CWR-specific oligo-painting technique holds broad applicability for chromosome engineering of numerous crops, as it allows rapid identification of alien chromosomes, reliable detection of homoeologous recombination, and visual tracking of the introgression process. It is promising to achieve directed and high-precision crop pre-breeding combined with other breeding techniques, such as CRISPR/Cas9-mediated chromosome engineering.

The Gossypium anomalum genome as a resource for cotton improvement and evolutionary analysis of hybrid incompatibility.

Cotton is an important crop that has been the beneficiary of multiple genome sequencing efforts, including diverse representatives of wild species for germplasm development. Gossypium anomalum is a wild African diploid species that harbors stress-resistance and fiber-related traits with potential application to modern breeding efforts. In addition, this species is a natural source of cytoplasmic male sterility and a resource for understanding hybrid lethality in the genus. Here, we report a high-quality de novo genome assembly for G. anomalum and characterize this genome relative to existing genome sequences in cotton. In addition, we use the synthetic allopolyploids 2(A2D1) and 2(A2D3) to discover regions in the G. anomalum genome potentially involved in hybrid lethality, a possibility enabled by introgression of regions homologous to the D3 (Gossypium davidsonii) lethality loci into the synthetic 2(A2D3) allopolyploid.

Whole-Genome Sequence of Synthesized Allopolyploids in Cucumis Reveals Insights into the Genome Evolution of Allopolyploidization.

The importance of allopolyploidy in plant evolution has been widely recognized. The genetic changes triggered by allopolyploidy, however, are not yet fully understood due to inconsistent phenomena reported across diverse species. The construction of synthetic polyploids offers a controlled approach to systematically reveal genomic changes that occur during the process of polyploidy. This study reports the first fully sequenced synthetic allopolyploid constructed from a cross between Cucumis sativus and C. hystrix, with high‐quality assembly. The two subgenomes are confidently partitioned and the C. sativus‐originated subgenome predominates over the C. hystrix‐originated subgenome, retaining more sequences and showing higher homeologous gene expression. Most of the genomic changes emerge immediately after interspecific hybridization. Analysis of a series of genome sequences from several generations (S0, S4–S13) of C. ×hytivus confirms that genomic changes occurred in the very first generations, subsequently slowing down as the process of diploidization is initiated. The duplicated genome of the allopolyploid with double genes from both parents broadens the genetic base of C. ×hytivus, resulting in enhanced phenotypic plasticity. This study provides novel insights into plant polyploid genome evolution and demonstrates a promising strategy for the development of a wide array of novel plant species and varieties through artificial polyploidization.

Analysis of Transcriptional Changes in Different Brassica napus Synthetic Allopolyploids.

Allopolyploidy is an evolutionary and mechanistically intriguing process involving the reconciliation of two or more sets of diverged genomes and regulatory interactions, resulting in new phenotypes. In this study, we explored the gene expression patterns of eight F2 synthetic Brassica napus using RNA sequencing. We found that B. napus allopolyploid formation was accompanied by extensive changes in gene expression. A comparison between F2 and the parent shows a certain proportion of differentially expressed genes (DEG) and activation\\silent gene, and the two genomes (female parent (AA)\\male parent (CC) genomes) showed significant differences in response to whole-genome duplication (WGD); non-additively expressed genes represented a small portion, while Gene Ontology (GO) enrichment analysis showed that it played an important role in responding to WGD. Besides, genome-wide expression level dominance (ELD) was biased toward the AA genome, and the parental expression pattern of most genes showed a high degree of conservation. Moreover, gene expression showed differences among eight individuals and was consistent with the results of a cluster analysis of traits. Furthermore, the differential expression of waxy synthetic pathways and flowering pathway genes could explain the performance of traits. Collectively, gene expression of the newly formed allopolyploid changed dramatically, and this was different among the selfing offspring, which could be a prominent cause of the trait separation. Our data provide novel insights into the relationship between the expression of differentially expressed genes and trait segregation and provide clues into the evolution of allopolyploids.

Genomic Asymmetry for Morphology in Allopolyploids Within and out of <i>Brassica</i>

The genomic asymmetry in the nonrandom retention and expression of controlling genes for some traits from one parental diploid is obvious in some natural and synthetic allopolyploids, and has the evolutionary implications. Here we review the genomic asymmetry for the morphological performance in three cultivated <i>Brassica</i> allotetraploids and some intergeneric allopolyploids within <i>Brassicaceae</i> species. For the phenotypic biases of <i>Brassica</i> allotetraploids, <i>Brassica oleracea</i> (genomes CC) is dominant over <i>B. nigra</i> (BB) and <i>B. rapa</i> (AA) in <i>B. carinata</i> (CCBB) and <i>B. napus</i> (CCAA), respectively, and <i>B. nigra</i> is dominant over <i>B. rapa</i> in <i>B. juncea</i> (BBAA), showing the C>B>A dominance hierarchy. However, the morphology of several <i>Brassica</i> species including <i>B. oleracea</i> at top dominance is largely recessive in their intergeneric hybrids and allopolyploids with other crucifers, such as <i>Raphanus sativus</i>, <i>Orychophragmus violaceus</i>. The morphology of <i>Arabidopsis thaliana </i>is also recessive in its two allotetraploids. Among the dominant features, the leaf serration is expressed consistently in these intergeneric and <i>Arabidopsis</i> allopolyploids. The phenotype expression of the recessive diploid is subject to the euploidy or aneuploidy state of its genome, and the dominant traits are still mostly expressed in the aneuploidy state of their genome. The morphological biases in these allopolyploids are discussed in the contexts of the genomic structure and interplay.

Pilar Catalán.

Pilar Catalán.

Hybrids between Brassica napus and B. nigra show frequent pairing between the B and A/C genomes and resistance to blackleg.

High frequencies of homoeologous and even non-homologous chromosome recombination in Brassica hybrids can transfer useful traits between genomes, but also destabilise synthetic allopolyploids. We produced triploid hybrids (2n = 3x = ABC) from the cross B. napus (rapeseed, 2n = 4x = AACC) × B. nigra (black mustard, 2n = 2x = BB) by embryo rescue and allohexaploid hybrids (2n = 6x = AABBCC = 54) by chromosome doubling of the triploids. These hybrids demonstrated resistance to blackleg disease (causal agent: Leptosphaeria maculans) inherited from their B. nigra parent. In order to assess the possibility of transfer of this resistance between the B genome and the A and C subgenomes of B. napus, as well as to assess the genomic stability of allohexaploids from the cross B. napus × B. nigra, frequencies of non-homologous chromosome pairing in these hybrids were assessed using classical cytogenetics and genomic in-situ hybridization. Meiosis was highly irregular, and non-homologous chromosome pairing between the B genome and the A/C genomes was common in both triploid hybrids (observed in 38% of pollen mother cells) and allohexaploid hybrids (observed in 15% of pollen mother cells). Our results suggest that introgression of blackleg resistance from the B genome into the A or C genomes should be possible, but that allohexaploids from this genome combination are likely unstable.

Cytogenetic Analyses Revealed Different Genome Rearrangement Footprints in Four ×BrassicoraphanusLines with Different Fertility Rates

×Brassicoraphanus (AARR, 2n = 38) is a synthetic intergeneric allopolyploid between Brassica rapa L. ssp. pekinensis (AA, 2n = 20) and Raphanus sativus L. var. rafiphera (RR, 2n = 18). Abnormalities in meiosis are main causes for infertility, especially in recent intergeneric allopolyploids. Several ×Brassicoraphanus lines showing varied fertility rates were produced previously, but no cytogenetic data specifying the reasons for infertility have been reported. In this study, we performed cytogenetic analyses in BB4, BB6, BB12, and BB50 lines to evaluate their chromosomal composition and behavior during meiosis. The four lines had relatively small chromosomes, ranging in length from 1.82 to 5.72 μm. BB6, BB12, and BB50 have euploid chromosome number of 2n = 38, whereas BB4 is an aneuploid with 2n - 1 = 37. Fluorescent in situ hybridization karyotype analysis by using 5S/45S rDNA revealed 5/7, 6/7, 5/5 and 5/5 pairs in BB4, BB6, BB12 and BB50, respectively. Genomic in situ hybridization analysis on cells in prophase I revealed varying frequencies of tetravalent pairing and sticky, ring, rod, and laggard chromosomes across the lines, which were more abundant in BB4 and BB6. Unlike BB4 and BB6, both BB12 and BB50 are known to have relatively higher seed fertility and uniform plant morphology. The varied degrees of chromosomal pairing stability during meiosis could explain the different fertility rates among the four ×Brassicoraphanus lines in this study. These data might facilitate breeding programs of ×Brassicoraphanus and further cytogenomic analyses.

Integrated QTL and eQTL Mapping Provides Insights and Candidate Genes for Fatty Acid Composition, Flowering Time, and Growth Traits in a F2 Population of a Novel Synthetic Allopolyploid Brassica napus.

Brassica napus (B. napus, AACC), is an economically important allotetraploid crop species that resulted from hybridization between two diploid species, Brassica rapa (AA) and Brassica olereacea (CC). We have created one new synthetic B. napus genotype Da-Ae (AACC) and one introgression line Da-Ol-1 (AACC), which were used to generate an F2 mapping population. Plants in this F2 mapping population varied in fatty acid content, flowering time, and growth-related traits. Using quantitative trait locus (QTL) mapping, we aimed to determine if Da-Ae and Da-Ol-1 provided novel genetic variation beyond what has already been found in B. napus. Making use of the genotyping information generated from RNA-seq data of these two lines and their F2 mapping population of 166 plants, we constructed a genetic map consisting of 2,021 single nucleotide polymorphism markers that spans 2,929 cM across 19 linkage groups. Besides the known major QTL identified, our high resolution genetic map facilitated the identification of several new QTL contributing to the different fatty acid levels, flowering time, and growth-related trait values. These new QTL probably represent novel genetic variation that existed in our new synthetic B. napus strain. By conducting genome-wide expression variation analysis in our F2 mapping population, genetic regions that potentially regulate many genes across the genome were revealed. A FLOWERING LOCUS C gene homolog, which was identified as a candidate regulating flowering time and multiple growth-related traits, was found underlying one of these regions. Integrated QTL and expression QTL analyses also helped us identified candidate causative genes associated with various biological traits through expression level change and/or possible protein function modification.

Whole Genome Incorporation and Epigenetic Stability in a Newly Synthetic Allopolyploid of Gynogenetic Gibel Carp.

Allopolyploidization plays an important role in speciation, and some natural or synthetic allopolyploid fishes have been extensively applied to aquaculture. Although genetic and epigenetic inheritance and variation associated with plant allopolyploids have been well documented, the relative research in allopolyploid animals is scarce. In this study, the genome constitution and DNA methylation inheritance in a newly synthetic allopolyploid of gynogenetic gibel carp were analyzed. The incorporation of a whole genome of paternal common carp sperm in the allopolyploid was confirmed by genomic in situ hybridization, chromosome localization of 45S rDNAs, and sequence comparison. Pooled sample-based methylation sensitive amplified polymorphism (MSAP) revealed that an overwhelming majority (98.82%) of cytosine methylation patterns in the allopolyploid were inherited from its parents of hexaploid gibel carp clone D and common carp. Compared to its parents, 11 DNA fragments in the allopolyploid were proved to be caused by interindividual variation, recombination, deletion, and mutation through individual sample-based MSAP and sequencing. Contrast to the rapid and remarkable epigenetic changes in most of analyzed neopolyploids, no cytosine methylation variation was detected in the gynogenetic allopolyploid. Therefore, the newly synthetic allopolyploid of gynogenetic gibel carp combined genomes from its parents and maintained genetic and epigenetic stability after its formation and subsequently seven successive gynogenetic generations. Our current results provide a paradigm for recurrent polyploidy consequences in the gynogenetic allopolyploid animals.

Homoeolog Expression Is Modulated Differently by Different Subgenomes in Brassica napus Hybrids and Allotetraploids

Synthetic and natural allotetraploid Brassica napus (2n = 38, AACC) have been widely used as a model to study the genetic changes associated with allopolyploidization; however, there has been little research on the homoeolog expression patterns and the roles of cis and trans regulation. Herein, homoeolog expression patterns were assessed by using RNA-seq for two interspecific hybrids (AnCo with the extracted A subgenome from natural B. napus, and ArCo with the A subgenome from extant B. rapa), synthetic and natural allopolyploids (CoCoArAr and AnAnCnCn), and the diploid parents. The ranges of homoeolog expression bias decreased after hybridization, whereas the extents of homoeolog expression bias and non-conserved expression, especially transgressive expression, increased over evolutionary time. Despite sharing the same C subgenome parent, these two hybrids showed different homolog expression patterns in many respects. In AnCo, the trans-regulatory factors from Co subgenome tended to cause downregulation of An subgenome homoeologs, but trans-regulatory factors from the An subgenome acted as both activators and repressors, and such asymmetric effects of trans-regulatory factors might explain why the homoeolog expression was biased toward the C subgenome after genome merger. No significant asymmetric effects of trans-regulatory factors were found in ArCo, which was consistent with the overall balanced expression of homoeologs. These results suggested that A subgenomes with different regulatory systems might act differently in modulating homoeolog expression after merger with the C subgenome, resulting in either balanced or unbalanced homoeolog expression biases.

Genetic identification of a newly synthetic allopolyploid strain with 206 chromosomes in polyploid gibel carp

A newly synthetic allopolyploid strain (SAS) was selected and established from gynogenetic offspring of gibel carp clone A+ with 156 chromosomes induced by common carp sperm with 50 chromosomes. In this study, the allopolyploid strain was detected to contain 206 chromosomes, and the growth trait was evaluated to have 25.15% growth faster than that of clone A+. Genetic marker analyses of transferrin (Tf) alleles, ITS1 sequences and mtDNA D-loop sequences indicated that the allopolyploid strain was synthetized from maternal gibel carp clone A+ and paternal common carp, and the synthetic chromosome sets were further confirmed by genomic in situ hybridization (GISH) and chromosome localization of 45S rDNA. Significantly, the synthetic allopolyploid strain has tended to be stable by five successive generations of gynogenesis, because it still keeps unisexual reproduction mode of gynogenesis. Therefore, it will become a novel variety for gibel carp aquaculture in future.

Genomic changes in generations of synthetic rapeseed-like allopolyploid grown under selection

Resynthesized Brassica napus L. is an important source for broadening genetic diversity and producing lines with desired characteristics. It is also a fine model to study the processes of genomic reorganizations in recently formed polyploids. We firstly performed molecular cytogenetic characterization of newly resynthesized rapeseed (B. rapa ssp. narinosa × B. oleracea ssp. capitata) and its parental species, and also examined genomic changes in hybrids of the succeeding generations grown under pressure of selection of yellow-seeded progeny. For karyotype studies, FISH/GISH with 45S, 5S rDNA, C genome specific BoB014O06 BAC clone and genomic DNA of parental B. rapa was performed. Synthetic S0–S2 hybrids had common rapeseed karyotypes (2n = 38) including 14 loci of 45S rDNA sites and 10 loci of 5S rDNA. Progeny selection led to gradual deletion of C genome chromosomes in hybrid karyotypes. So, in karyotypes of S6 and S7 hybrids, the chromosome number was reduced to 2n = 20–22, and only chromosomes of A genome bearing 10–13 loci of 45S rDNA and 8–10 loci of 5S rDNA, variations in chromosome number, chromosome rearrangements as well as examples of trisomy and monosomy were revealed. Our findings indicate an enhanced genome instability in resynthesized rapeseed lines developed under the pressure of selection which might lead to chromosome rearrangements or/and deletions and even elimination of the whole parental genome in hybrids in succeeding generations. The approach can be useful for the development of rapeseed lines with trisomy, chromosome addition/substitution lines important for genetics and plant breeding.