Allotriploid Hybrids Research Articles

Biomass yields, reproductive fertility, compositional analysis, and genetic diversity of newly developed triploid giant miscanthus hybrids

AbstractMiscanthus ×giganteus (giant miscanthus), first found as a naturally occurring hybrid, has shown promise as a bioenergy/biomass crop throughout much of the temperate world. This allotriploid (2n = 3x = 57) hybrid resulted from a cross between tetraploid Miscanthus sacchariflorus (2n = 4x = 76) and diploid Miscanthus sinensis (2n = 2x = 38) and is particularly desirable due to its low fertility that minimizes reseeding and potential invasiveness. However, there is limited genetic diversity in commonly grown cultivars of triploid M. ×giganteus and breeding and development efforts to improve and domesticate this crop have been minimal. Here, we report on newly developed M. ×giganteus hybrids compared with the industry standard M. ×giganteus '1993‐1780'. Dry biomass yields of new hybrids ranged from 19.5 to 32.4 Mg/ha/year for the fourth growing season, compared with 21.0 Mg/ha/year for M. ×giganteus '1993‐1780'. Plant reproductive fertility remained low for all accessions with overall fertility [(seed set × seed germination)/100] ranging from 0.3% to 4.5% for new hybrids compared to 0.4% for M. ×giganteus '1993‐1780'. Culm density and height varied among accessions and were positively correlated with increased biomass. Based on compositional analyses, theoretical ethanol yields ranged from 9, 740 to 16,278 L/ha/year for new hybrids compared to 10,406 L/ha/year for M. ×giganteus '1993‐1780'. Relative feed value indices were low overall and ranged between 66.0 and 72.8 for new hybrids compared to M. ×giganteus '1993‐1780' with 71.3. The genetic diversity of new hybrids, compared with existing cultivars, was characterized using whole genome sequences. Based on pair‐wise distances, cluster analysis clearly showed increased diversity of new hybrids compared with earlier selections. These results document new triploid hybrids of M. ×giganteus with enhanced biomass and theoretical ethanol yields in combination with broader genetic diversity and lowreproductive fertility.

Transcriptome Shock in Developing Embryos of a Brassica napus and Brassica rapa Hybrid.

Interspecific crosses that fuse the genomes of two different species may result in overall gene expression changes in the hybrid progeny, called 'transcriptome shock'. To better understand the expression pattern after genome merging during the early stages of allopolyploid formation, we performed RNA sequencing analysis on developing embryos of Brassica rapa, B. napus, and their synthesized allotriploid hybrids. Here, we show that the transcriptome shock occurs in the developing seeds of the hybrids. Of the homoeologous gene pairs, 17.1% exhibit expression bias, with an overall expression bias toward B. rapa. The expression level dominance also biases toward B. rapa, mainly induced by the expression change in homoeologous genes from B. napus. Functional enrichment analysis revealed significant differences in differentially expressed genes (DEGs) related to photosynthesis, hormone synthesis, and other pathways. Further study showed that significant changes in the expression levels of the key transcription factors (TFs) could regulate the overall interaction network in the developing embryo, which might be an essential cause of phenotype change. In conclusion, the present results have revealed the global changes in gene expression patterns in developing seeds of the hybrid between B. rapa and B. napus, and provided novel insights into the occurrence of transcriptome shock for harnessing heterosis.

Chromosome-level genome assembly of a triploid poplar Populus alba 'Berolinensis'.

Many recent studies have provided significant insights into polyploid breeding, but limited research has been carried out on trees. The genomic information needed to understand growth and response to abiotic stress in polyploidy trees is largely unknown, but has become critical due to the threats to forests imposed by climate change. Populus alba 'Berolinensis,' also known "Yinzhong poplar," is a triploid poplar from northeast China. This hybrid triploid poplar is widely used as a landscape ornamental and in urban forestry owing to its adaptation to adverse environments and faster growth than its parental diploid. It is an artificially synthesized male allotriploid hybrid, with three haploid genomes of P.alba 'Berolinensis' originating from different poplar species, so it is attractive for studying polyploidy genomic mechanisms in heterosis. In this study, we focused on the allelic genomic interactions in P.alba 'Berolinensis,' and generated a high-quality chromosome-level genome assembly consisting of 19 allelic chromosomes. Its three haploid chromosome sets are polymorphic with an average of 25.73 nucleotide polymorphism sites per kilobase. We found that some stress-related genes such as RD22 and LEA7 exhibited sequence differences between different haploid genomes. The genome assembly has been deposited in our polyploid genome online analysis website TreeGenomes (https://www.treegenomes.com). These polyploid genome-related resources will provide a critical foundation for the molecular breeding of P.alba 'Berolinensis' and help us uncover the allopolyploidization effects of heterosis and abiotic stress resistance and traits of polyploidy species in the future.

The dynamics of lncRNAs transcription in interspecific F1 allotriploid hybrids between Brassica species

Interspecific hybridization is the intrinsic forces behind genome evolution. Long non-coding RNAs (lncRNAs) are important for plant biological processes regulation. However, it is unclear that these non-coding fractions are impacted by interspecific hybridization. Here we examined the profiles of lncRNAs by comparing them with coding genes in Brassica napus, three accessions of Brassica rapa, and their F1 hybrids. 6206 high-confidential lncRNAs were identified in F 1 hybrids and their parentals, and the lncRNAs transcriptome in the F1 hybrids was reprogrammed by the genome shock. Notably, genome-wide unbalanced of lncRNAs were observed between An and Ar subgenomes, ELD (Expression Level Dominance) was biased toward the An -genome in F1 hybrids, and ELD of non-conserved lncRNAs was more than conserved lncRNAs. Our findings demonstrate that the reprogramed lncRNAs acts as important role in enhancing plant plasticity, leading to the acquisition of desirable traits in polyploid Brassica species.

A Novel Allotriploid Hybrid Derived From Female Goldfish × Male Bleeker’s Yellow Tail

Hybridization is a traditional and effective strategy to alter the genotypes and phenotypes of the offspring, and distant hybridization is a useful strategy to generate polyploids in fish. In this study, goldfish (Carassius auratus, GF, 2n = 100) and Bleeker’s yellow tail (Xenocypris davidi Bleeker, YT, 2n = 48), which belong to different subfamilies, were crossed with each other. The cross of female GF × male YT successfully obtained hybrid offspring (GFYT hybrids), while the cross of female YT × male GF was lethal, and all the fertilized eggs stopped developing before the neurula stage of embryogenesis. All GFYT hybrids possessed 124 chromosomes (3n = 124) with two sets from GF and one set from YT. The measurable and countable traits of GFYT hybrids were identified, and the genetic characteristics of 5S rDNA between GFYT hybrids and their parents were also revealed. There were, respectively, four and three different 5S rDNA types in GF (assigned as GF-Ⅰ∼Ⅳ) and YT (assigned as YT-Ⅰ∼Ⅲ), and GFYT hybrids specifically inherited YT-Ⅰ and YT-Ⅱ 5S rDNA types from YT and GF-Ⅲ and GF-Ⅳ from GF. In addition, there were only testis-like and fat-like gonads been found in GFYT hybrids. Interestingly, there were pyknotic and heteromorphous chromatin and invaginated cell membrane observed in the spermatids of testis-like gonads, but no mature sperm were found. Furthermore, TUNEL assays indicated that, compared with control, apparent apoptotic signals, which were mainly distributed around spermatid regions, were detected in the testis-like gonads, and the expression of apoptosis pathway-related genes including p53, bcl-2, bax, and caspase9 was significantly upregulated. Moreover, the expression of meiosis-related genes including spo11, dmc1, and rad51 showed an abnormally high expression, but mns1 and meig1, two key genes involved in the maturation of spermatid, were extremely downregulated. In brief, this is the first report of allotriploid via distant hybridization between GF and YT that possessing different chromosome numbers in vertebrates. The obtainment of GFYT hybrids not only harbors potential benefits and application in aquaculture but also further extends the understanding of the influence of hybridization and polyploidization on the genomic constitution of the hybrid offspring. Furthermore, they can be used as a model to test the origin and consequences of polyploidization and served as a proper resource to study the underlying mechanisms of spermatogenesis dysfunctions.

Transcriptome shock in interspecific F1 allotriploid hybrids between Brassica species.

Interspecific hybridization drives the evolution of angiosperms and can be used to introduce novel alleles for important traits or to activate heterosis in crop breeding. Hybridization brings together gene expression networks from two different species, potentially causing global alterations of gene expression in the F1 plants which is called 'transcriptome shock'. Here, we explored such a transcriptome shock in allotriploid Brassica hybrids. We generated interspecific F1 allotriploid hybrids between the allotetraploid species Brassica napus and three accessions of the diploid species Brassica rapa. RNA-seq of the F1 hybrids and the parental plants revealed that 26.34-30.89% of genes were differentially expressed between the parents. We also analyzed expression level dominance and homoeolog expression bias between the parents and the F1 hybrids. The expression-level dominance biases of the Ar, An, and Cn subgenomes was genotype and stage dependent, whereas significant homoeolog expression bias was observed among three subgenomes from different parents. Furthermore, more genes were involved in trans regulation than in cis regulation in allotriploid F1 hybrids. Our findings provide new insights into the transcriptomic responses of cross-species hybrids and hybrids showing heterosis, as well as a new method for promoting the breeding of desirable traits in polyploid Brassica species.

A Modified Meiotic Recombination in Brassica napus Largely Improves Its Breeding Efficiency

Simple SummaryThe selection of varieties more resilient to disease and climate change requires generating new genetic diversity for breeding. The main mechanism for reshuffling genetic information is through the recombination of chromosomes during meiosis. We showed in oilseed rape (Brassica napus, AACC, 2n = 4x = 38), which is a natural hybrid formed from a cross between turnip (B. rapa, AA, 2n = 2x = 20) and cabbage (B. oleracea, CC, 2n = 2x = 18), that there is significantly more crossovers occurring along the entire A chromosomes in allotriploid AAC (crossbetween B. napus and B. rapa) than in diploid AA or allotetraploid AACC hybrids. We demonstrated that these allotriploid AAC hybrids are highly efficient to introduce new variability within oilseed rape varieties, notably by enabling the introduction of small genomic regions carrying genes controlling agronomically interesting traits.Meiotic recombination is the main tool used by breeders to generate biodiversity, allowing genetic reshuffling at each generation. It enables the accumulation of favorable alleles while purging deleterious mutations. However, this mechanism is highly regulated with the formation of one to rarely more than three crossovers, which are not randomly distributed. In this study, we showed that it is possible to modify these controls in oilseed rape (Brassica napus, AACC, 2n = 4x = 38) and that it is linked to AAC allotriploidy and not to polyploidy per se. To that purpose, we compared the frequency and the distribution of crossovers along A chromosomes from hybrids carrying exactly the same A nucleotide sequence, but presenting three different ploidy levels: AA, AAC and AACC. Genetic maps established with 202 SNPs anchored on reference genomes revealed that the crossover rate is 3.6-fold higher in the AAC allotriploid hybrids compared to AA and AACC hybrids. Using a higher SNP density, we demonstrated that smaller and numerous introgressions of B. rapa were present in AAC hybrids compared to AACC allotetraploid hybrids, with 7.6 Mb vs. 16.9 Mb on average and 21 B. rapa regions per plant vs. nine regions, respectively. Therefore, this boost of recombination is highly efficient to reduce the size of QTL carried in cold regions of the oilseed rape genome, as exemplified here for a QTL conferring blackleg resistance.

An allotriploid hybrid developed from a female 2n gamete was obtained from Eriobotrya japonica (Thunb.) Lindl × E. deflexa (Hemsl.) Nakai

A triploid was obtained from loquat (Eriobotrya japonica cv. ‘Ninghaibai’) (2n = 2x = 34) × bronze loquat (Eriobotrya deflexa, 2n = 2x = 34). The leaves of the triploid had no petioles, similar to the female parent, and its stipules were obvious and dark green, similar to those of the male parent. There were signals of male parent DNA on ten chromosomes of the triploid according to genomic in situ hybridization (GISH) when DNA of bronze loquat was used as probe; the diploid hybrid also had ten chromosomes with signals of male parent DNA. All five selected simple sequence repeats (SSR) primers amplified specific loci of both parents. In particular, primer CH04c06 amplified both specific loci of the female parent but only amplified one specific locus of the male parent. The results of GISH and SSR marker analysis indicated that the 2n gamete which formed the triploid was from first-division restitution or pre-meiotic doubling of the female parent cv. ‘Ninghaibai’. The allotriploid can be used to breed seedless loquat germplasm with different flowering times from loquat directly or to breed new germplasm containing a few genomic components from bronze loquat, such as monosomic alien addition lines, monosomic alien substitution lines, and introgression lines. This is the first report of allotriploid in the genus Eriobotrya and it will provide a basis for breeding new loquat germplasms by distant hybridization and provide an ideal material for breeding new loquat varieties with different flowering time or time to maturity.

Gene flow between diploid and tetraploid junipers - two contrasting evolutionary pathways in two Juniperus populations

BackgroundGene flow and polyploidy have been found to be important in Juniperus evolution. However, little evidence has been published elucidating the association of both phenomena in juniper taxa in the wild. Two main areas were studied in Spain (Eastern Iberian Range and Sierra de Baza) with both diploid and tetraploid taxa present in sympatry. Gene flow and ploidy level were assessed for these taxa and the resulted offspring.ResultsTwenty-two allo-triploid hybrids between J. sabina var. sabina and J. thurifera were found in the Eastern Iberian Range population. However, in the Sierra de Baza population no triploids were found. Instead, 18 allo-tetraploid hybrids between two tetraploid taxa: J. sabina var. balkanensis and J. thurifera were discovered. High genetic diversity was exhibited among the tetraploid hybrids at Sierra de Baza, in contrast to the genetically identical triploid hybrids at the Eastern Iberian Range; this suggests meiotic difficulties within the triploid hybrids. In addition, unidirectional gene flow was observed in both studied areas.ConclusionPolyploidy and hybridization can be complementary partners in the evolution of Juniperus taxa in sympatric occurrences. Juniperus was shown to be an ideal coniferous model to study these two phenomena, independently or in concert.

Differences in immune components of blood, spleen and head kidney between diploid and auto- and allotriploid Salmonidae

Immune components were investigated in peripheral blood and in spleen and head kidney of autotriploid Salmo trutta f. lacustris, Salvelinus fontinalis, and Salvelinus umbla, and of allotriploid hybrids of S. trutta f. lacustris x Onchorynchus mykiss and S. fontinalis x O. mykiss in comparison to their diploid parents. In peripheral blood the number of lymphocytes was reduced in all investigated autotriploids and in the allotriploid S. trutta f. lacustris x O.mykiss, and the numbers of thrombocytes in autotriploid S. trutta f. lacustris and in both allotriploids. Alternative pathway of complement activity and immunoglobulin concentration were significantly decreased in all investigated auto- and allotriploids, lysozyme activity in autotriploid S. fontinalis and in both allotriploids. In the spleen of the 3 autotriploids the number of erythrocytes was increased, while the number of lymphoid precursor cells was decreased. In their head kidney the erythrocytes numbers were decreased and the numbers of erythropoietic precursor cells and the melanomacrophage centers were increased. Contrary, cytology of spleen and head kidney of the two allotriploid hybrids was similar to diploid controls. Caspase 1, caspase 6, lysozyme, and acid phosphatase activity and immunoglobulin concentration of spleen and head kidney showed specific changes which were related to cytological results. These data indicate alterations in immune system and in lymphoid organs of auto- and allotriploid Salmonidae.

First evidence for allotriploid hybrids between Juniperus thurifera and J. sabina in a sympatric area in the French Alps

Key message Discovery of the first case of allotriploid juniper in a wild population in the French Alps where the parental species occurs in sympatry. ContextInterspecific hybridization and polyploidy are important evolutionary phenomena in vascular plants. Natural hybridization between species living in sympatry can sometimes occur. Less frequent are successful hybridizations between species having different ploidy levels. At Saint Crépin location (French Alps), where sympatry between the tetraploid Juniperus thurifera and the diploid Juniperus sabina occurs, three individuals with an atypical morphology have been observed.AimsProspect interspecific hybrids and interspecific genetic introgression occurrence.MethodsFlow cytometry was employed to screen ploidy levels. Four chloroplast markers, nrDNA (ITS), and AFLP markers were used to unravel hybridization and potential introgression events. Variability of pollen size and morphology was assessed to have a first insight on the regularity of microsporogenesis in hybrids.ResultsThe three putative hybrids were shown to be triploids. Molecular data demonstrated that these individuals were hybrids originated from a cross between J. sabina and J. thurifera and suggested that a backcross at least with J. thurifera is possible. Male triploid hybrids produced heterogeneous pollen and displayed evidence of irregularity in the microsporogenesis.ConclusionThis study sheds light on a rare case of hybridization in a natural sympatric population of two Juniperus species with different ploidy levels. This mechanism might have been an important driver for the evolution and diversification of this coniferous genus.

Developmental potential of somatic and germ cells of hybrids betweenCarassius auratusfemales andHemigrammocypris rasborellamales

The cause of hybrid sterility and inviability has not been analyzed in the fin-fish hybrid, although large numbers of hybridizations have been carried out. In this study, we produced allo-diploid hybrids by cross-fertilization between female goldfish (Carassius auratus) and male golden venus chub (Hemigrammocypris rasborella). Inviability of these hybrids was due to breakage of the enveloping layer during epiboly or due to malformation with serious cardiac oedema around the hatching stage. Spontaneous allo-triploid hybrids with two sets of the goldfish genome and one set of the golden venus chub genome developed normally and survived beyond the feeding stage. This improved survival was confirmed by generating heat-shock-induced allo-triploid hybrids that possessed an extra goldfish genome. When inviable allo-diploid hybrid cells were transplanted into goldfish host embryos at the blastula stage, these embryos hatched normally, incorporating the allo-diploid cells. These allo-diploid hybrid cells persisted, and were genetically detected in a 6-month-old fish. In contrast, primordial germ cells taken from allo-diploid hybrids and transplanted into goldfish hosts at the blastula stage had disappeared by 10 days post-fertilization, even under chimeric conditions. In allo-triploid hybrid embryos, germ cells proliferated in the gonad, but had disappeared by 10 weeks post-fertilization. These results showed that while hybrid germ cells are inviable even in chimeric conditions, hybrid somatic cells remain viable.

Production of allohexaploid Brassica hybrid between tuber mustard (Brassica juncea L. var. crassicaulis Chen &Yang) and Chinese kale (Brassica oleracea var. alboglabra Bailey)

Brassica is a key agricultural genus, containing oilseed and vegetable crops, which includes three major genomes (A, B and C), distributed in diploid (genomes: AA, BB and CC) and allotetraploid species (genomes: AABB, AACC and BBCC). Interspecific hybridization between Brassica species is an important way to combine valuable characters from the different species. In the present study, distant hybridization between inbred lines of tuber mustard (Brassica juncea var. crassicaulis Chen &Yang; AABB) and Chinese kale (Brassica oleracea var. alboglabra Bailey; CC) was conducted and allotriploid interspecific hybrids were identified. The hybrid seed set was improved by gibberellin treatment of the stigmas prior to pollination. Eight hybrid seedlings were obtained by bud pollination and ovary culture, and all seedlings were confirmed to be true hybrids by phenotypic analysis, cytological observation (27 chromosomes) and the use of parent-specific randomly amplified polymorphic DNA (RAPD) molecular markers. After tissue culture and colchicine-induced chromosome-doubling treatment, allohexaploid hybrids (AABBCC) were isolated and identified from the eight hybrids. The chromosomes of five plants were completely doubled, with a doubling rate of 5.68 %, and the number of chimeric plants was 24, at a rate of 27.27 %, when roots were immersed in 0.1 % colchicine solution. In comparison with the allotriploid hybrid, the synthetic hexaploid hybrids showed unique phenotypes with normal anthers, well-developed pods, larger flowers and petals, thicker stems than either parent and other phenotype types intermediate between the two parents. In the present study, synthetic tri-genomic Brassica vegetable germplasm, combining the A, B and C genomes, was created for use in vegetable breeding, as well as for a basis for further investigation of the mechanisms of Brassica evolution and adaptation. The current study is of great significance for the use of these allohexaploid plants to explore the heterotic effects of distant hybridization and for genetic improvement.

Cytological atlas at meiosis reveals insights into pollen fertility in synthetic Brassica allotriploids between allotetraploid B. carinata and diploid B. rapa

The formation of allopolyploid crops basically depends on the successful interspecific hybridization and polyploidization, which generally involves in a combination of distinct but related genomes from independent species. But cytological analysis of these initially synthesized allohaploids immediately after genome merging is poorly explored in regards to anther and pollen development to date. In this study, Brassica trigenomic allohaploids (ABC) were produced to investigate the immediate effects of the genome combinations on pollen fertility during anther development via crosses between natural allotetraploid B. carinata (BBCC) and diploid B. rapa (AA). The results showed that in the synthetic Brassica allotriploids (ABC), the anther development was completely disrupted, and the pollen grains were mostly inviable with varied genetic complements. In addition, the meiosis course was aberrantly altered and eccentric chromosomal configurations including multivalent, bridges and lags occurred frequently during metaphase I to anaphase II. Genomic in situ hybridization (GISH) further revealed that B genome of homoeology was frequently apt to interact with A and C genomes, and cytoskeletal organizations was improperly distributed during meiosis in these synthetic Brassica allotriploids. Furthermore, we also confirmed that the expression of typical meiosis-related genes was obviously repressed during anther development in these Brassica allotriploids. Taken together, our results provide a detailed cytology for insights into pollen development in the synthetic allotriploid hybrids, which are conventionally considered as a useful genetic resource for polyploid Brassica breeding.

Sub-Genome Polyploidization Effects on Metabolomic Signatures in Triploid Hybrids of Populus

Allopolyploids are known to have superior advantages such as high growth speed. Triploids have even greater heterozygosity, explaining more phenotypic variance than 2n hybrid F1 and have therefore become new resources in breeding. To date, the metabolomic basis underlying polyploidization vigor remains unclear. Here, we identified and compared 235 metabolites in the shoot apical buds between multiple allo-triploid populations and parental 2n hybrid F1 in Populus via metabolome profiling using liquid chromatography–mass spectrometry (LC–MS) assays. Associations with growth vigor in three types of allo-triploid populations, namely first division restitution (FDR), second division restitution (SDR) and postmeiotic restitution (PMR) generated from doubled 2n female gametes and male gametes of 2n hybrid, were also investigated. Each allo-triploid population has different sub-genome duplicated. Major metabolomes were amino acids, secondary metabolism associated, and carbohydrates. We mapped 181 metabolites into known metabolism pathways in the Kyoto Encyclopedia of Genes and Genomes (KEGG). Ten compounds, i.e., fructose 1,6-diphosphate and xylulose, were more abundant in all allo-triploids than the 2n hybrid. Principal component analysis revealed the abundance of metabolites fell into distinct clusters corresponding to ploidy composition. Heterozygosity in triploids mainly effected the contents of carbohydrates and secondary metabolites rather than lipids. Comparisons between subgroups with different growth rates revealed some carbohydrates and secondary metabolites of flavonoids were positively associated with gene expression and the high growth vigor. The results provided an informative metabolomic basis for factors conferring growth vigor in polyploid Populus.

The Sterility of Allotriploid Fish and Fertility of Female Autotriploid Fish.

Based on the formation of an autotetraploid fish line (4nAUT, 4n = 200; F2–F11) derived from the distant hybridization of female Carassius auratus red var. (RCC, 2n = 100) × male Megalobrama amblycephala (BSB, 2n = 48), we produced autotriploid hybrids (3nAUT) by crossing females of RCC with males of 4nAUT and allotriploid hybrids (3nALT) by crossing females of Cyprinus carpio (CC, 2n = 100) with males of 4nAUT. The aim of this study was to comparatively investigate the reproductive characteristics of 3nALT and 3nAUT. We investigated morphological traits, chromosomal numbers, DNA content and gonadal development in 3nAUT and 3nALT. The results indicated both 3nAUT and 3nALT possessed 150 chromosomes and were triploid hybrids. The females and males of 3nALT and males of 3nAUT had abnormal gonadal development and could not generate mature eggs or sperm, but the females of 3nAUT had normal gonadal development and generated mature eggs at 2 years old. The females of 3nAUT generated different sizes of eggs, which fertilized with haploid sperm from RCC and formed viable diploid, triploid, and tetraploid offspring. The formation of these two kinds of triploid hybrids provides an ideal model for studying the reproductive traits of triploid hybrids, which is of great value in animal genetics and reproductive biology.

Pairing analysis and in situ Hybridisation reveal autopolyploid-like behaviour in Solanum commersonii\xa0\xd7\xa0S. tuberosum (potato) interspecific hybrids

Wild potato relatives are rich sources of desirable traits for introgressive hybridisation into cultivated potato. One of them, Solanum commersonii (2n = 2x = 24, 1EBN, endosperm balance number), is an important species belonging to the potato tertiary genepool. It can be used in potato breeding through bridge crosses and 2n gamete production. Triploid F1 hybrids between S. commersonii (through spontaneous 2n egg formation) and diploid 2EBN Solanum tuberosum Group Phureja were crossed with S. tuberosum Group Tuberosum, resulting in successive backcross 1, 2 and 3 progenies. The main aim of this study was to determine if there are any barriers to homoeologous pairing and recombination in the allotriploid (S. commersonii × potato) hybrids and their backcrosses, and so to predict if S. commersonii chromosomes can be transmitted to the next generation and introgressed into their recipient potato chromosomes. Microscopic observations of spread pollen mother cells suggested no preferential pairing in the triploid hybrids, while chromosome transmission and segregation in further meiotic stages were fairly balanced. Fluorescent in situ hybridisation with BAC probes (BAC FISH) was used to obtain markers to trace the meiotic behaviour of specific chromosome pairs. Moreover, genomic in situ hybridisation (GISH) demonstrated no obvious differences in fluorescence signals between the homoeologues suggesting that repetitive sequences did not diverge much between the parental species. As a consequence, we were not able to trace the course of the S. commersonii chromosomes in the successive introgressive hybridisation backcross generations. Our results strongly point at a high genomic similarity between the homoeologous chromosomes promising high suitability of S. commersonii in introgressive hybridisation breeding of potato.

Amplifying recombination genome-wide and reshaping crossover landscapes in Brassicas.

Meiotic recombination by crossovers (COs) is tightly regulated, limiting its key role in producing genetic diversity. However, while COs are usually restricted in number and not homogenously distributed along chromosomes, we show here how to disrupt these rules in Brassica species by using allotriploid hybrids (AAC, 2n = 3x = 29), resulting from the cross between the allotetraploid rapeseed (B. napus, AACC, 2n = 4x = 38) and one of its diploid progenitors (B. rapa, AA, 2n = 2x = 20). We produced mapping populations from different genotypes of both diploid AA and triploid AAC hybrids, used as female and/or as male. Each population revealed nearly 3,000 COs that we studied with SNP markers well distributed along the A genome (on average 1 SNP per 1.25 Mbp). Compared to the case of diploids, allotriploid hybrids showed 1.7 to 3.4 times more overall COs depending on the sex of meiosis and the genetic background. Most surprisingly, we found that such a rise was always associated with (i) dramatic changes in the shape of recombination landscapes and (ii) a strong decrease of CO interference. Hybrids carrying an additional C genome exhibited COs all along the A chromosomes, even in the vicinity of centromeres that are deprived of COs in diploids as well as in most studied species. Moreover, in male allotriploid hybrids we found that Class I COs are mostly responsible for the changes of CO rates, landscapes and interference. These results offer the opportunity for geneticists and plant breeders to dramatically enhance the generation of diversity in Brassica species by disrupting the linkage drag coming from limits on number and distribution of COs.

Overcoming sterility and unilateral incompatibility of Solanum lycopersicum × S. sitiens hybrids

The wild tomato relative Solanum sitiens is a rare endemic plant of the Atacama Desert of Chile. This species exhibits unique traits—tolerance to drought, salinity and low temperatures, resistances to certain pathogens, and modified fruit ripening—that could be of interest to tomato breeders. However, strong reproductive barriers, including hybrid inviability, sterility, and unilateral incompatibility (UI), prevent or impede introgression with cultivated tomato. Using embryo culture, we obtained a large number of F1 S. lycopersicum × S. sitiens hybrids, and evaluated their fertility and compatibility as male or female parents. Diploid hybrids were functionally male-sterile and strongly rejected pollen of S. lycopersicum by UI. Chromosome doubling produced fertile allotetraploids, from which allotriploids (two genomes of S. lycopersicum, one of S. sitiens) were derived by crossing to diploid tomato. Rejection of tomato pollen occurred lower in styles of allotriploid than diploid hybrids. The pollen compatibility factors ui1.1 and ui6.1 were not sufficient for compatibility on S. sitiens allotriploid hybrids, unlike comparable hybrids with S. lycopersicoides. Pollen from F1 S. lycopersicum × S. pennellii was used to overcome UI on pistils of diploid S. lycopersicum × S. sitiens hybrids. This cross yielded progeny that were fertile and compatible in direct backcrosses to S. lycopersicum. These results establish a feasible method for transferring the S. sitiens genome into cultivated tomato.

Genomic composition of microspore-derived plants from a Chinese cabbage (Brassica rapa ssp. pekinensis) × heading cabbage (B. oleracea var. capitata) allotriploid (ACC) hybrid based on genomic in situ hybridization

SummaryGenomic in situ hybridisation (GISH) was used to investigate the chromosome composition of five microsporederived plants previously obtained from a Chinese cabbage (Brassica rapa ssp. pekinesis) _ heading cabbage (B. oleracea var. capitata) allotriploid hybrid (ACC). Chromosome numbers and compositions varied between the microspore-derived plants examined. Plant ‘GB-17’ contained 17 chromosomes, of which ten and seven were derived from B. oleracea (genome C) and B. rapa (genome A), respectively. Plant ‘GB-18’ contained 18 chromosomes, including nine C and nine A chromosomes. Plant ‘GB-19’ was an allodiploid plant that contained 19 chromosomes, including nine C and ten A chromosomes. Plant ‘GB-20’ contained 20 chromosomes, including eight C and 12 A chromosomes. Plant ‘GB-26’ contained 26 chromosomes, including 12 C and 14 A chromosomes. Recombinant chromosomes were observed in plants ‘GB-17’ and ‘GB-20’. The morphological features and meiotic behaviour of plant ‘GB-19’ were further examined and compared with the two parental plants. Chromosomal homoeology between the A and C genomes was revealed. Our results provide credible evidence to identify the different chromosomes between the two genomes. In addition, GISH was shown to be a powerful tool with which to characterise the genomic composition of hybrids and to reveal chromosome recombination events during hybridisation.