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Summary: Ultraviolet (UV) radiation influences development and genome stability in organisms; however, its impact on meiosis, a special cell division essential for the delivery of genetic information across generations in eukaryotes, has not yet been elucidated.In this study, by performing cytogenetic studies, we reported that UV radiation does not damage meiotic chromosome integrity but attenuates centromere‐mediated chromosome stability and induces unreduced gametes in Arabidopsis thaliana.We showed that functional centromere‐specific histone 3 (CENH3) is required for obligate crossover formation and plays a role in the protection of sister chromatid cohesion under UV stress. Moreover, we found that UV specifically alters the orientation and organization of spindles and phragmoplasts at meiosis II, resulting in meiotic restitution and unreduced gametes. We determined that UV‐induced meiotic restitution does not rely on the UV Resistance Locus8‐mediated UV perception and the Tapetal Development and Function1‐ and Aborted Microspores‐dependent tapetum development, but possibly occurs via altered JASON function and downregulated Parallel Spindle1.This study provides evidence that UV radiation influences meiotic genome stability and gametophytic ploidy consistency in flowering plants. [ABSTRACT FROM AUTHOR]

Premise: Quaking aspen is a clonal tree species that has mixed ploidy, often with high relative abundance of both diploids and triploids but no haploids or tetraploids. Triploids typically have low fertility, leaving their occurrence apparently unlikely from an evolutionary perspective, unless they provide a "triploid bridge" to generating higher‐fitness tetraploids—which are not observed in this species. This study focused on how triploidy can be maintained in quaking aspen. Methods: A computational model was used to simulate gamete production, sexual reproduction, asexual reproduction, parent survival, and offspring survival in a population. All parameters were assumed to be cytotype‐dependent and environment‐independent. Sampling methods were used to identify parameter combinations consistent with observed cytotype frequencies. Results: Many processes and parameter values were sufficient to yield a moderate frequency of triploids, and very few were necessary. The most plausible route involved higher triploid survival at the parent or offspring stage and limited unreduced gamete production by either diploid or triploid parents. Triploid fertility was helpful but not necessary. Conclusions: The coexistence of diploids and triploids in quaking aspen is statistically likely and promoted by the existence of commonly observed, long‐lived triploid clones. However, other mechanisms not captured by the model related to environmental variation could also occur. Further empirical data or more complex but difficult‐to‐parameterize models are needed to gain further insight. [ABSTRACT FROM AUTHOR]

SUMMARY: Through the inactivation of genes that act during meiosis it is possible to direct the genetic make‐up of plants in subsequent generations and optimize breeding schemes. Offspring may show higher recombination of parental alleles resulting from elevated crossover (CO) incidence, or by omission of meiotic divisions, offspring may become polyploid. However, stable mutations in genes essential for recombination, or for either one of the two meiotic divisions, can have pleiotropic effects on plant morphology and line stability, for instance by causing lower fertility. Therefore, it is often favorable to temporarily change gene expression during meiosis rather than relying on stable null mutants. It was previously shown that virus‐induced gene silencing (VIGS) can be used to transiently reduce CO frequencies. We asked if VIGS could also be used to modify other processes throughout meiosis and during pollen formation in Arabidopsis thaliana. Here, we show that VIGS‐mediated knock‐down of FIGL1, RECQ4A/B, OSD1 and QRT2 can induce (i) an increase in chiasma numbers, (ii) unreduced gametes and (iii) pollen tetrads. We further show that VIGS can target both sexes and different genetic backgrounds and can simultaneously silence different gene copies. The successful knock‐down of these genes in A. thaliana suggests that VIGS can be exploited to manipulate any process during or shortly after meiosis. Hence, the transient induction of changes in inheritance patterns can be used as a powerful tool for applied research and biotechnological applications. Significance Statement: The present work demonstrates that VIGS is an efficient tool to quickly accomplish breeding aims such as an increase in crossover recombination, the generation of polyploid offspring and modification of pollen‐specific processes. [ABSTRACT FROM AUTHOR]

Common wheat (Triticum aestivum L., AABBDD genome) is thought to have emerged through natural hybridization between Triticum turgidum L. (AABB genome) and Aegilops tauschii Coss. (DD genome). Hybridization barriers and doubling of the trihaploid F1 hybrids' genome (ABD) via unreduced gamete fusion had key roles in the process. However, how T. turgidum, the maternal progenitor, was involved in these mechanisms remains unknown. An artificial cross‐experiment using 46 cultivated and 31 wild T. turgidum accessions and a single Ae. tauschii tester with a very short genetic distance to the common wheat D genome was conducted. Cytological and quantitative trait locus analyses of F1 hybrid genome doubling were performed. The crossability and ability to cause hybrid inviability did not greatly differ between the cultivars and wild accessions. The ability to cause hybrid genome doubling was higher in the cultivars. Three novel T. turgidum loci for hybrid genome doubling, which influenced unreduced gamete production in F1 hybrids, were identified. Cultivated T. turgidum might have increased the probability of the emergence of common wheat through its enhanced ability to cause genome doubling in F1 hybrids with Ae. tauschii. The ability enhancement might have involved alterations at a relatively small number of loci. [ABSTRACT FROM AUTHOR]

Although reproductive assurance has been suggested to be one of the most important factors shaping the differential distributional patterns between sexuals and asexuals (geographic parthenogenesis), it has only rarely been studied in natural populations of vascular plants with autonomous apomixis. Moreover, there are almost no data concerning the putative relationship between the level of apomictic versus sexual plant reproduction on one hand, and reproductive assurance on the other.We assessed the level of sexual versus apomictic reproduction in diploid and triploid plants of Hieracium alpinum across its distributional range using flow cytometric analyses of seeds, and compared the level of potential and realized seed set, i.e. reproductive assurance, between the two cytotypes under field and greenhouse conditions.Flow cytometric screening of embryos and endosperms of more than 4,100 seeds showed that diploids produced solely diploid progeny sexually, while triploids produced triploid progeny by obligate apomixis. Potential fruit set was much the same in diploids and triploids from the field and the greenhouse experiment. While in the pollination‐limited environment in the greenhouse apomictic triploids had considerably higher realized fruit set than sexual diploids, there was no significant difference between cytotypes under natural conditions. In addition, sexuals varied to a significantly larger extent in realized fruit set than asexuals under both natural and greenhouse conditions.Our results indicate that triploid plants reproduce by obligate apomixis, assuring more stable and predictable fruit reproduction when compared to sexual diploids. This advantage could provide apomictic triploids with a superior colonisation ability, mirrored in a strong geographic parthenogenesis pattern observed in this species. [ABSTRACT FROM AUTHOR]

Natural hybridisation and polyploidy are currently recognised as drivers of biodiversity, despite early scepticism about their importance. The Mediterranean region is a biodiversity hotspot where geological and climatic events have created numerous opportunities for speciation through hybridisation and polyploidy. Still, our knowledge on the frequency of these mechanisms in the region is largely limited, despite both phenomena are frequently cited in studies of Mediterranean plants. We reviewed information available from biodiversity and cytogenetic databases to provide the first estimates of hybridisation and polyploidy frequency in the Mediterranean region. We also inspected the most comprehensive modern Mediterranean Flora ( Flora iberica) to survey the frequency and taxonomic distribution of hybrids and polyploids in Iberian Peninsula. We found that <6% of Mediterranean plants were hybrids, although a higher frequency was estimated for the Iberian Peninsula (13%). Hybrids were concentrated in few families and in even fewer genera. The overall frequency of polyploidy (36.5%) was comparable with previous estimates in other regions; however our estimates increased when analysing the Iberian Peninsula (48.8%). A surprisingly high incidence of species harbouring two or more ploidy levels was also observed (21.7%). A review of the available literature also showed that the ecological factors driving emergence and establishment of new entities are still poorly studied in the Mediterranean flora, although geographic barriers seem to play a major role in polyploid complexes. Finally, this study reveals several gaps and limitations in our current knowledge about the frequency of hybridisation and polyploidy in the Mediterranean region. The obtained estimates might change in the future with the increasing number of studies; still, rather than setting the complete reality, we hope that this work triggers future studies on hybridisation and polyploidy in the Mediterranean region. [ABSTRACT FROM AUTHOR]

Fertilization involving unreduced (2 n) gametes is considered the dominant mechanism of polyploid formation in angiosperms; however, our knowledge of the prevalence of and evolutionary mechanisms maintaining 2 n gametes in natural populations is limited. We hypothesize that 2 n gametes are deleterious consequences of meiotic errors maintained by mutation-selection balance and should increase in species with relaxed opportunities for selection on sexual processes (asexuality), reduced efficacy of selection (asexuality, selfing) and increased genome instability (high chromosome number)., We used flow cytometry to estimate male 2 n gamete production in 60 populations from 24 species of Brassicaceae. We quantified variation in 2 n gamete production within and among species, and examined associations with life history, reproductive mode, genome size and chromosomal number while accounting for phylogeny., Most individuals produced < 2% 2 n male gametes, whereas a small number had > 5% (up to 85%) production. Variation in 2 n gamete production was significant among species and related to reproductive system; asexual species produced significantly more 2 n gametes than mixed-mating and outcrossing species., Our results, unique in their multi-species perspective, are consistent with 2 n gametes being deleterious but maintained when opportunities for selection are limited. Rare individuals with elevated 2 n gamete production may be key contributors to polyploid formation. [ABSTRACT FROM AUTHOR]

The value of flow cytometry for quantifying unreduced (2N) pollen production in plants is well recognized; however, the approach has been limited by technical obstacles to obtaining high quality nuclei fluorescence histograms and the difficulty in distinguishing 2N nuclei from 1N doublets. Here, we use mathematical arguments and observations of fluorescence properties of angiosperm pollen nuclei to generate guidelines for applying pulse analysis to correct for doublets in pollen nuclei data. We show that the theoretical requirements for applying pulse analysis for doublet correction are met when nucleus fluorescence height and/or width measures in the unreduced gamete (2C DNA content) region exhibit bimodality (reflecting singlets and doublets) in combination with unimodal distributions of the same parameters in the reduced gamete (1C) region. These conditions are regularly met in the family Brassicaceae but not in the Asteraceae and Poaceae. We further show that when these requirements are met, pulse analysis estimates of doublet proportions are well correlated with estimates obtained with microscopy. We propose guidelines for doublet correction when estimating frequencies of unreduced male gametes. © 2015 International Society for Advancement of Cytometry [ABSTRACT FROM AUTHOR]

Premise of the study. Most plants are polyploid and have more than two copies of the genome. The evolutionary success of polyploids is often attributed to their potential to harbor increased genetic variation, but it is poorly understood how polyploids can attain such variation. Because of their formation bottleneck, newly formed tetraploids start out with little variation. Tetraploids may attain genetic variation through a combination of new mutations, recurrent formation, and gene exchange with diploid ancestors or related tetraploid species. We explore the role of gene exchange and introgression in autotetraploid Rorippa amphibia, a species that harbors more genetic variation than its diploid ancestors. Methods: We crossed autotetraploid R. amphibia to diploid conspecifics and tetraploid R. sylvestris and backcrossed resulting F1 hybrids. We used flow cytometry to determine the ploidy of all progeny. Key results: Tetraploids of R. amphibia and R. sylvestris were interfertile; F1 hybrids were fertile and could backcross. Crosses between diploids and tetraploids yielded a small number of viable, often tetraploid progeny. This indicates that unreduced gametes can facilitate gene flow from diploids to tetraploids. We detected a frequency of unreduced gametes of around 2.7 per 1000, which was comparable between diploids and tetraploids. Conclusions: Introgression from tetraploid R. sylvestris provides a realistic source of variation in autotetraploid R. amphibia. Only in a scenario where other compatible partners are absent, for example immediately after tetraploidization, gene flow through unreduced gametes from diploids could be an important source of genetic variation for tetraploids. [ABSTRACT FROM AUTHOR]

Ploidy manipulations are achieved by utilizing unreduced gametes, somatic chromosome doubling or haploidization. Apomixis, the asexual mode of reproduction through seeds, involves two of these phenomena viz., apomeiosis (unreduced embryo-sac formation) and parthenogenesis (fertilization-independent embryogenesis). The two components when partitioned may yield high frequencies of triploids and haploids, and thus, serve as a tool to manipulate ploidy levels when appropriately supplemented with hybridization schemes. Utilizing this ybridization-supplemented pomixis-components artitioning pproach (HAPA), the largest ploidy series in a crop plant was generated in guinea grass ( Panicum maximum Jacq.). Eighty-nine diverse guinea grass accessions were characterized for their reproductive capacities to identify tetraploid (2 n = 4 x = 32) accession(s) with high apospory but reduced parthenogenetic capacity, thereby producing a high frequency of hexaploid seeds. Seeds from accession IG 04-164 were utilized to produce a ploidy series by appropriately combining partitioned apomixis components and hybridizations in two seasonal cycles. We successfully obtained plants representing ploidies 3 x, 4 x, 5 x, 6 x, 8 x and 9 x, all derived from a single 4 x progenitor. Production methodology, reproducibility and utilization of HAPA in cytogenetic and molecular studies are discussed. [ABSTRACT FROM AUTHOR]

The wheat progenitors and other wild relatives continue to be important sources of genes for agronomically desirable traits, which can be transferred into durum wheat ( Triticum turgidum; 2 n = 4 x = 28; AABB genomes) cultivars via hybridization. Chromosome pairing in durum × alien species hybrids provides an understanding of genomic relationships, which is useful in planning alien gene introgression strategies. Two durum cultivars, ‘Lloyd’ and ‘Langdon’, were crossed with diploid wheatgrass, Thinopyrum bessarabicum (2 n = 2 x = 14; JJ), to synthesize F1 hybrids (2 n = 3 x = 21; ABJ) with Ph1. ‘Langdon’ disomic substitution 5D(5B) was used as a female parent to produce F1 hybrids without Ph1, which resulted in elevation of pairing between durum and grass chromosomes – an important feature from the breeding standpoint. The F1 hybrids were backcrossed to respective parental cultivars and BC1 progenies were raised. ‘Langdon’ 5D(5B) substitution × Th. bessarabicum F1 hybrids were crossed with normal ‘Langdon’ to obtain BC1 progeny. Chromosome pairing relationships were studied in F1 hybrids and BC1 progenies using both conventional staining and fluorescent genomic in situ hybridization (fl-GISH) techniques. Multicolour fl-GISH was standardized for characterizing the nature and specificity of chromosome pairing: A–B, A–J and B–J pairing. The A–J and B–J pairing will facilitate gene introgression in durum wheat. Multicolour fl-GISH will help in characterizing alien chromosome segments captured in the durum complement and in their location in the A and/or B genome, thereby accelerating chromosome engineering research. [ABSTRACT FROM AUTHOR]

Abstract The multiploid mutant of durum wheat is a genotype that produces unreduced gametes. Our objective was to test the recovery of pentaploid hybrids in crosses of the mutant with rye and Triticum monococcum L. Compared with check crosses, the mutant had a two-third reduction in percent seed set for rye crosses, but had only a slight decrease in crossability with T. monococcum . Pentaploid hybrids were associated with plump seeds of the mutant/rye cross, and with shrivelled seeds of the mutant/T. monococcum cross. We suggest that the endosperm balance number hypothesis explains the association of pentaploid hybrids with endosperm type. This association made for easy recovery of pentaploid hybrids from crosses to both species. Mature, plump seeds from the mutant/rye cross were germinated and pentaploid hybrids were recovered. One pentaploid hybrid was recovered for every 50.5 and 15.1 florets pollinated with rye and T. monococcum , respectively. Unreduced gametes in the multiploid mutant will facilitate interspecific hybridization by reducing the time to produce pentaploid plants. [ABSTRACT FROM AUTHOR]

Successful hybridization between Lilium concolor and Lilium longiforum has not been reported but ovary slice culture technique, after cut-style pollination has now been used to produce diploid and triploid interspecific hybrids between these species. Reciprocal crosses between diploid cultivars (2n = 2x = 24) were conducted. 5, 10, 15, 20, 25, 30, 35, 40, and 45 days after pollination (DAP), ovaries were sliced and cultured on a modified Murashige and Skoog (MS) medium without growth regulators and NH[sub4]NO[sub3], supplemented with 6% sucrose, 50mg/I yeast extract and 0.25% gelrite at pH 6.3. For the L. concolor x L. longiflorum cross, embryo germination was found to be best at 20 DAP, while for the L. longiflorum x L concolor at 25 DAP. After transfer to a MS (half-strength) medium supplemented with 1.5% sucrose, 0.25% gelrite and 0.2% active charcoal at pH 5.8, diploid and triploid hybrid plants were developed. All regenerated plants were identified as hybrids on the basis of karyotype and isozyme analyses. Ovary slice culture technique as a method of producing polyploids is discussed. [ABSTRACT FROM AUTHOR]

Self-sterile interspecific hybrids between diploid Lolium perenne and either Festuca arundinacea or F. gigantea (both hexaploids) set some seeds when subjected to a heavy pollen cloud from the amphiploids derived from the hybrids. All the progeny were octoploid and were formed from the fertilization of unreduced gametes in the F[SUB1] hybrid by amphiploid pollen. The significance of the results in relation to breeding efforts within the Lolium/Festuca complex is discussed. [ABSTRACT FROM AUTHOR]