Postzygotic Mechanisms Research Articles

Yam (Dioscorea rotundata Poir.) Displays Prezygotic and Postzygotic Barriers to Prevent Autogamy in Monoecious Cultivars

Cultivated yam (D. rotundata) is a staple tuber crop in West Africa whose sexual reproduction control remains largely unknown despite its importance for plant breeding programs. In this paper, we compared self-pollination, intracultivar cross-pollination and intercultivar cross-pollination in three monoecious cultivars (Amoula, Heapala and Yassi). Results showed that pollen viability (49%) and stigma receptivity (40%) were similar in monoecious and dioecious plants, suggesting that autogamy could occur in monoecious plants. However, fruit and seed sets were significantly lower after self-pollination compared to cross-pollination. Overall, autogamy reached 11% and pollen lability was almost zero (<1%). The low percentage of pollen grains germinating on the stigma (37%) and pollen tubes reaching the ovules (25%) after self-pollination partly explained the low seed set. Strong inbreeding depression was observed after self-pollination and almost all fruits and about 75% of the seeds resulting from self-pollination showed malformations. Seed germination was also 20 times lower after self-pollination compared to cross-pollination. Sexual reproduction remained low in D. rotundata even after cross-pollination as fruit and seed set did not exceed 18% and 13% respectively. Moreover, comparison between intracultivar cross-pollination and self-pollination revealed intravarietal genetic diversity inside the analyzed yam cultivars. Overall, our results showed that D. rotundata has a very low tolerance to autogamy in monoecious cultivars and has developed pre- and postzygotic mechanisms to limit selfing.

Pre- and postzygotic mechanisms preventing hybridization in co-occurring species of the Impatiens purpureoviolacea complex.

In the species‐rich genus Impatiens, few natural hybrids are known, even though closely related species often occur sympatrically. In this study, we aim to bridge the gap between micro‐ and macro‐evolution to disentangle pre‐ and postzygotic mechanisms that may prevent hybridization in the Impatiens purpureoviolacea complex from Central Africa. We analyzed habitat types, species distribution, pollination syndromes, pollinator dependency, genome sizes, and chromosome numbers of seven out of the ten species of the complex as well as of one natural hybrid and reconstructed the ancestral chromosome numbers of the complex. Several species of the complex occur in sympatry or geographically very close to each other. All of them are characterized by pre‐ and/or postzygotic mechanisms potentially preventing hybridization. We found four independent polyploidization events within the complex. The only known natural hybrid always appears as single individual and is self‐fertile. But the plants resulting from self‐pollinated seeds often die shortly after first flowering. These results indicate that the investigated mechanisms in combination may effectively but not absolutely prevent hybridization in Impatiens and probably occur in other genera with sympatric species as well.

Rapid Divergence of Key Spermatogenesis Genes in nasuta-Subgroup of Drosophila.

The crosses between closely relatedDrosophilaspecies usually produce sterile hybrid males with spermatogenesis disrupted at post-meiotic phase, especially in sperm individualization stage than the pre-meiotic stage. This is possibly due to the rapid interspecies divergence of male sex and reproduction-related genes. Here we annotated 11 key spermatogenesis genes in 35 strains of species belonging tonasuta-subgroup ofDrosophila, where many interspecies crosses produce sterile males. We characterized the divergence and polymorphism in the protein coding regions by employing gene-wide, codon-wide, and lineage-specific selection analysis to test the mode and strength of selection acting on these genes.Our analysis showed signature of positive selection at bag of marbles (bam)andbenign gonial cell neoplasma (bgcn)despite the selection constrains and the absence of endosymbiont infection which could potentially drive rapid divergence due to an arms race whileroughex (rux)showed lineage-specific rapid divergence in frontal sheen complex ofnasuta-subgroup.cookie monster (comr)showed rapid divergence consistent with the possibility of meiotic arrest observed in sterile hybrids ofDrosophilaspecies. Rapid divergence observed atdon juan (dj) and Mst98Ca-likewas consistent with fused sperm-tail abnormality observed in the hybrids ofDrosophila nasutaandDrosophila albomicans. These findings highlight the potential role of rapid nucleotide divergence in bringing about hybrid incompatibility in the form of male sterility; however, additional genetic manipulation studies can widen our understanding of hybrid incompatibilities. Furthermore, our study emphasizes the importance of young species belonging tonasuta-subgroup ofDrosophilain studying post-zygotic reproductive isolation mechanisms.

Genotype-by-environment interactions drive the maintenance of genetic variation in a Salmo trutta L. hybrid zone.

Allopatric gene pools can evolve in different directions through adaptive and nonadaptive processes and are therefore a source of intraspecific diversity. The connection of these previously isolated gene pools through human intervention can lead to intraspecific diversity loss, through extirpation of native populations or hybridization. However, the mechanisms leading to these situations are not always explicitly documented and are thus rarely used to manage intraspecific diversity. In particular, genotype‐by‐environment (GxE) interactions can drive postzygotic reproductive isolation mechanisms that may result in a mosaic of diversity patterns, depending on the local environment. We test this hypothesis using a salmonid species (Salmo trutta) in the Mediterranean (MED) area, where intensive stocking from non‐native Atlantic (ATL) origins has led to various outcomes of hybridization with the native MED lineage, going from MED resilience to total extirpation via full hybridization. We investigate patterns of offspring survival at egg stage in natural environments, based on parental genotypes in interaction with river temperature, to detect potential GxE interactions. Our results show a strong influence of maternal GxE interaction on embryonic survival, mediated by maternal effect through egg size, and a weak influence of paternal GxE interaction. In particular, when egg size is large and temperature is cold, the survival rate of offspring originating from MED females is three times higher than that of ATL females’ offspring. Because river temperatures show contrast at small scale, this cold adaptation for MED females’ offspring constitutes a potent postzygotic mechanism to explain small‐scale spatial heterogeneity in diversity observed in MED areas where ATL fish have been stocked. It also indicates that management efforts could be specifically targeted at the environments that actively favor native intraspecific diversity through eco‐evolutionary processes such as postzygotic selection.

Genetic Mapping of the HLA1 Locus Causing Hybrid Lethality in Nicotiana Interspecific Hybrids.

Hybrid lethality, a postzygotic mechanism of reproductive isolation, is a phenomenon that causes the death of F1 hybrid seedlings. Hybrid lethality is generally caused by the epistatic interaction of two or more loci. In the genus Nicotiana, N. debneyi has the dominant allele Hla1-1 at the HLA1 locus that causes hybrid lethality in F1 hybrid seedlings by interaction with N. tabacum allele(s). Here, we mapped the HLA1 locus using the F2 population segregating for the Hla1-1 allele derived from the interspecific cross between N. debneyi and N. fragrans. To map HLA1, several DNA markers including random amplified polymorphic DNA, amplified fragment length polymorphism, and simple sequence repeat markers, were used. Additionally, DNA markers were developed based on disease resistance gene homologs identified from the genome sequence of N. benthamiana. Linkage analysis revealed that HLA1 was located between two cleaved amplified polymorphic sequence markers Nb14-CAPS and NbRGH1-CAPS at a distance of 10.8 and 10.9 cM, respectively. The distance between these markers was equivalent to a 682 kb interval in the genome sequence of N. benthamiana.

No evidence for incipient speciation by selfing in North American Arabidopsis lyrata.

Self-fertilization inherently restricts gene flow by reducing the fraction of offspring that can be produced by inter-population matings. Therefore, mating system transitions from outcrossing to selfing could result in reproductive isolation between selfing and outcrossing lineages and provide a starting point for speciation. In newly diverged lineages, for example after a transition to selfing, further reproductive isolation can be caused by a variety of prezygotic and post-zygotic mechanisms that operate before, during and after pollination. In animals, prezygotic barriers tend to evolve faster than post-zygotic ones. This is not necessarily the case for plants, for which the relative importance of post-mating, post-fertilization and early-acting post-zygotic barriers has been investigated far less. To test whether post-pollination isolation exists between populations of North American Arabidopsis lyrata that differ in breeding (self-incompatible versus self-compatible) and mating system (outcrossing versus selfing), we compared patterns of seed set after crosses made within populations, between populations of the same mating system and between populations with different mating systems. We found no evidence for post-pollination isolation between plants from selfing populations (self-compatible, low outcrossing rates) and outcrossing populations (self-incompatible, high outcrossing rates) via either prezygotic or early-acting post-zygotic mechanisms. Together with the results of other studies indicating the absence of reproductive barriers acting before and during pollination, we conclude that the transition to selfing in this study system has not led to the formation of reproductive barriers between selfing and outcrossing populations of North American A.lyrata.

Incomplete reproductive isolation and strong transcriptomic response to hybridization between sympatric sister species of salmon.

Global change is altering ecosystems at an unprecedented rate. The resulting shifts in species ranges and reproductive timing are opening the potential for hybridization between closely related species which could dramatically alter the genetic diversity, adaptive capacity and evolutionary trajectory of interbreeding taxa. Here, we used behavioural breeding experiments, in vitro fertilization experiments, and whole-transcriptome gene expression data to assess the potential for and consequences of hybridization between Chinook and Coho salmon. We show that behavioural and gametic prezygotic barriers between socio-economically valuable Chinook and Coho salmon are incomplete. Postzygotically, we demonstrate a clear transcriptomic response to hybridization among F1 Chinook-Coho offspring. Genes transgressively expressed within hybrids were significantly enriched with genes encoded in the nucleus but localized to the mitochondrion, suggesting a potential role for mito-nuclear incompatibilities as a postzygotic mechanism of hybrid breakdown. Chinook and Coho salmon are expected to continue to respond to climate change with shifts in migration timing and habitat use, potentiating hybridization between these species. The downstream consequences of hybridization on the future of these threatened salmon, and the ecosystems they inhabit, is unknown.

The role of the chromosomal rearrangements in the evolution and speciation of Elopiformes fishes (Teleostei; Elopomorpha)

Elopiformes is one of the most ancient orders of Teleostei fishes and includes the Megalopidae and Elopidae families. Although relevant to scientific research, cytogenetic and molecular data are lacking for both families. Here, we compared Elopiformes species from the perspective of genetic divergence and cytogenetics to elucidate the chromosomal diversification and evolutionary processes of Elopiformes. The cytogenetic results showed that Megalops atlanticus and Elops smithi are 2n = 50, with differences in their chromosomal formulas. The distributions of heterochromatin and repetitive DNA sequences revealed many characteristics shared between Elopiformes and other basal lineages of Teleostei fishes. In addition, the results suggested that different chromosomal rearrangements have occurred in the analysed Elopiformes species. The species tree indicated that Megalopidae and Elopidae were monophyletic groups, but the genetic divergence was higher for Megalops species than for Elops species. The small divergence and sympatric distribution of Elops species may not prevent hybridization. However, the karyotype differentiation may be an effective post-zygotic mechanism preventing continuous hybridization, and it could be the key factor in this speciation process. In conclusion, this is the first report of Elopiformes species that integrates molecular and cytogenetic data. Our results emphasize the importance of cytogenetic data for further defining Elopidae species.

Asymmetric Fitness of Second-Generation Interspecific Hybrids Between Ciona robusta and Ciona intestinalis.

Reproductive isolation is central to speciation, but interspecific crosses between two closely related species can produce viable and fertile hybrids. Two different species of tunicates in the same ascidian genus, Ciona robusta and Ciona intestinalis, can produce hybrids. However, wild sympatric populations display limited gene flow, suggesting the existence of obstacles to interspecific reproduction that remain unknown. Here, we took advantage of a closed culture system to cross C. robusta with C. intestinalis and established F1 and F2 hybrids. We monitored post-embryonic development, survival, and sexual maturation to characterize the genetic basis of simple traits, and further probe the physiological mechanisms underlying reproductive isolation. Partial viability of first and second generation hybrids suggested that both pre- and postzygotic mechanisms contributed to genomic incompatibilities in hybrids. We observed asymmetric fitness, whereby the C. intestinalis maternal lines fared more poorly in our system, pointing to maternal origins of species-specific sensitivity. We discuss the possibility that asymmetrical second generation inviability and infertility emerge from interspecific incompatibilities between the nuclear and mitochondrial genomes, or other maternal effect genes. This work paves the way to quantitative genetic approaches to study the mechanisms underlying genomic incompatibilities and other complex traits in the genome-enabled Ciona model.

Evolutionary dynamics of pre- and postzygotic reproductive isolation in cichlid fishes.

Cichlid fishes are exceptionally species-rich, speciated at explosive rates and, hence, are a model system in speciation research. Yet, their reproductive isolating barriers have, so far, not been comprehensively studied. Here, we review current knowledge on pre- and postzygotic mechanisms in cichlids. While premating isolation is the norm in cichlids, its strength varies across lineages and with the geographical setting. Moreover, manipulations of ambient conditions tended to reduce assortative mating among closely related species, suggesting that premating isolation in cichlids is often fragile and context dependent. The observed lack of complete reproductive isolation is supported by past and present hybridization events that have contributed to diversity by creating novel allelic combinations. On the other hand, our meta-analysis highlights that intrinsic postzygotic isolation might accumulate faster than assumed. Mild forms of genetic incompatibilities, such as sex ratio distortion, can already be observed among closely related species. Therefore, cessation of gene flow by strong reproductive isolation in cichlids requires a combination of premating prezygotic isolation supplemented with intrinsic and extrinsic postzygotic barriers. Further, we suggest crucial next steps to improve our knowledge about reproductive barriers in cichlids to understand the evolutionary dynamics of pre- and postzygotic isolation mechanisms during adaptive radiations. This article is part of the theme issue 'Towards the completion of speciation: the evolution of reproductive isolation beyond the first barriers'.

Symbionts do not affect the mating incompatibility between the Brazilian-1 and Peruvian morphotypes of the Anastrepha fraterculus cryptic species complex

The South American fruit fly, Anastrepha fraterculus, is clearly undergoing a speciation process. Among others, two of their morphotypes, the Brazilian-1 and Peruvian, have accumulated differences in pre- and post-zygotic mechanisms resulting in a degree of reproductive isolation. Both harbor a different strain of Wolbachia, which is a widespread endosymbiotic bacterium among many invertebrates producing a range of reproductive effects. In this paper, we studied the role of this bacterium as one of the factors involved in such isolation process. Infected and cured laboratory colonies were used to test pre- and post-zygotic effects, with special emphasis in uni- and bi-directional cytoplasmic incompatibility (CI). We showed that Wolbachia is the only known reproductive symbiont present in these morphotypes. Wolbachia reduced the ability for embryonic development in crosses involving cured females and infected males within each morphotype (uni-directional CI). This inhibition showed to be more effective in the Peruvian morphotype. Bi-directional CI was not evidenced, suggesting the presence of compatible Wolbachia strains. We conclude that Wolbachia is not directly involved in the speciation process of these morphotypes. Other mechanisms rather than CI should be explored in order to explain the reduced mating compatibility between the Brazilian-1 and Peruvian morphotypes.

Effects of temperature and salinity on juveniles of two ascidians, one native and one invasive, and their hybrids

Human-mediated transports of species, i.e. biological introductions, promote secondary contacts between previously spatially isolated species. When the species are not reproductively isolated, hybridization is an important possible outcome of these secondary contacts, influencing the fate of both the native and non-native species. Such a situation is exemplified by the introduction of the warm-temperate tunicate Ciona robusta, native to Asia, into the European native range of its cold-temperate congener Ciona intestinalis. Despite an old divergence (ca. 4Mya), the two species lack reproductive isolation with fertile F1-hybrids produced in laboratory. However, a recent population genomic study showed that hybrids are extremely rare in the wild and back-crossed individuals are absent, suggesting the existence of reproductive barriers between the two species in nature. Post-zygotic mechanisms like selection against hybrids by the environment were thus hypothesized. Because the two species have only been recently reclassified, data on their respective ecology and their hybrids are lacking. To investigate these environmental effects, we carried out 21 experimental crosses and examined survival and individual growth rate of ca. 1900 F1-juveniles produced with intra- and inter-specific crosses. Four treatments combining two temperatures (12°C and 17°C) and two salinity levels (25 and 35), representative of conditions found in nature, were applied to one-week old juveniles. After 28days, a high survival rate was observed in the three F1 categories, with no difference between hybrids and parental species. As expected based on biogeography studies and field monitoring of the two species, the results showed that C. robusta performed particularly well in warm temperature with standard salinity condition. F1-hybrids did not show outbreeding depression, with growth rates similar to or higher than F1-C. intestinalis. Altogether the two species performed well under the tested environmental conditions, and post-zygotic ecological effects alone are unlikely to be effective against hybridization between the native and non-native species.

The Reproductive Ecology of Diploid and Tetraploid Galax urceolata

Polyploidy is common in nature, particularly among plants, and is often associated with significant morphological, ecological, and evolutionary change. These changes may contribute to prezygotic and postzygotic isolation between cytotypes; therefore, polyploidy is commonly thought to lead to speciation. Recent work suggests prezygotic mechanisms may be more important in determining isolation than postzygotic mechanisms. We compared the reproductive ecology of populations of diploid and tetraploid Galax to assess the potential for gene flow between the cytotypes. Galax urceolata is native to the southern Appalachian Mountains and is a natural polyploid series with diploid, triploid, and autotetraploid individuals occurring in uniform and mixed-cytotype populations. We found the temporal peak of flower production in tetraploids slightly preceded that of diploids; however, the overall distributions of flowering phenologies were similar. Hymenoptera and Diptera were the most common floral visitors and diploids were visited slightly more often than tetraploids; however, there were no differences between cytotypes in terms of the taxonomic composition of their floral visitors. Seed production is likely pollen limited, but again this does not differ between cytotypes. While tetraploids produced more flowers than diploids, there was no difference in fruit production. Finally, our data suggest that Galax urceolata is self-incompatible. In sum we found few differences in the reproductive ecology of diploid and tetraploid Galax, which when combined with obligate outcrossing increases the potential for gene flow between cytotypes and reduces the likelihood of differentiation.

Pollinators and crossability as reproductive isolation barriers in two sympatric oil-rewarding Calceolaria (Calceolariaceae) species

Pollinator species are widely accepted as an important factor in plant reproductive isolation. Although mostly investigated in plants visited by different groups of pollinators (e.g., hummingbirds vs bees), few studies have examined the role of pollinators belonging to the same taxonomic group (e.g., only bees) on plant reproductive isolation. In this study, we investigate this question by evaluating pre- and post-zygotic mechanisms putatively involved in the reproductive isolation of two oil-rewarding sympatric Calceolaria species (i.e., Calceolaria filicaulis and C. arachnoidea) in an Andean ecosystem of Chile. We estimated reproductive isolation values using a combination of field (pollinator visitation rates) and experimental (intra and interspecific manual cross-pollination and seed germination of parents and hybrids) evidence. The two Calceolaria species were preferentially visited by different oil-collecting bee species. Results from hand cross-pollination experiments indicate that intraspecific crossings produced significantly more seeds than interspecific ones. Notwithstanding, seed germination essays did not reveal differences between parental and hybrids. Taken together, these results suggest that pollinator species are responsible for most of the reproductive isolation in the two Calceolaria species studied here. This study is the first assessment of pollinator-mediated reproductive isolation in Calceolaria species and the first to document reproductive barriers in oil-rewarding plants.

Assessing pre- and post-zygotic barriers between North Atlantic eels (Anguilla anguilla and A. rostrata).

Elucidating barriers to gene flow is important for understanding the dynamics of speciation. Here we investigate pre- and post-zygotic mechanisms acting between the two hybridizing species of Atlantic eels: Anguilla anguilla and A. rostrata. Temporally varying hybridization was examined by analyzing 85 species-diagnostic single-nucleotide polymorphisms (SNPs; FST ⩾0.95) in eel larvae sampled in the spawning region in the Sargasso Sea in 2007 (N=92) and 2014 (N=460). We further investigated whether genotypes at these SNPs were nonrandomly distributed in post-F1 hybrids, indicating selection. Finally, we sequenced the mitochondrial ATP6 and nuclear ATP5c1 genes in 19 hybrids, identified using SNP and restriction site associated DNA (RAD) sequencing data, to test a previously proposed hypothesis of cytonuclear incompatibility leading to adenosine triphosphate (ATP) synthase dysfunction and selection against hybrids. No F1 hybrids but only later backcrosses were observed in the Sargasso Sea in 2007 and 2014. This suggests that interbreeding between the two species only occurs in some years, possibly controlled by environmental conditions at the spawning grounds, or that interbreeding has diminished through time as a result of a declining number of spawners. Moreover, potential selection was found at the nuclear and the cytonuclear levels. Nonetheless, one glass eel individual showed a mismatch, involving an American ATP6 haplotype and European ATP5c1 alleles. This contradicted the presence of cytonuclear incompatibility but may be explained by that (1) cytonuclear incompatibility is incomplete, (2) selection acts at a later life stage or (3) other genes are important for protein function. In total, the study demonstrates the utility of genomic data when examining pre- and post-zyotic barriers in natural hybrids.

Ecological speciation in sympatric palms: 2. Pre- and post-zygotic isolation.

We evaluated reproductive isolation in two species of palms (Howea) that have evolved sympatrically on Lord Howe Island (LHI, Australia). We estimated the strength of some pre‐ and post‐zygotic mechanisms in maintaining current species boundaries. We found that flowering time displacement between species is consistent across in and ex situ common gardens and is thus partly genetically determined. On LHI, pre‐zygotic isolation due solely to flowering displacement was 97% for Howea belmoreana and 80% for H. forsteriana; this asymmetry results from H. forsteriana flowering earlier than H. belmoreana and being protandrous. As expected, only a few hybrids (here confirmed by genotyping) at both juvenile and adult stages could be detected in two sites on LHI, in which the two species grow intermingled (the Far Flats) or adjacently (Transit Hill). Yet, the distribution of hybrids was different between sites. At Transit Hill, we found no hybrid adult trees, but 13.5% of younger palms examined there were of late hybrid classes. In contrast, we found four hybrid adult trees, mostly of late hybrid classes, and only one juvenile F1 hybrid in the Far Flats. This pattern indicates that selection acts against hybrids between the juvenile and adult stages. An in situ reciprocal seed transplant between volcanic and calcareous soils also shows that early fitness components (up to 36 months) were affected by species and soil. These results are indicative of divergent selection in reproductive isolation, although it does not solely explain the current distribution of the two species on LHI.

Mosaic deletion of 20pter due to rescue by somatic recombination

We report on a unique case of a mosaic 20pter-p13 deletion due to a somatic repair event identified by allele differentiating single nucleotide polymorphism (SNP) probes on chromosomal microarray. Small terminal deletions of 20p have been reported in a few individuals and appear to result in a variable phenotype. This patient was a 24-month-old female who presented with failure to thrive and speech delay. Chromosomal microarray analysis (CMA) performed on peripheral blood showed a 1.6 Mb deletion involving the terminus of 20p (20pter-20p13). This deletion appeared mosaic by CMA and this suspicion was confirmed by fluorescence in situ hybridization (FISH) analysis. Additionally, the deletion interval at 20p was directly adjacent to 15 Mb of mosaic copy-neutral loss of heterozygosity (LOH). The pattern of SNP probes was highly suggestive of a somatic repair event that resulted in rescue of the deleted region using the non-deleted homologue as a template. Structural mosaicism is rare and most often believed to be due to a postzygotic mechanism. This case demonstrates the additional utility of allele patterns to help distinguish mechanisms and in this case identified the possibility of either a post-zygotic repair of a germline deletion or a post-zygotic deletion with somatic recombination repair in a single step.

Reproductive isolation between sympatric sister species, Mussaenda kwangtungensis and M. pubescens var. alba.

Reproductive isolation defines the biological species concept and plays a key role in the formation and maintenance of species. The relative contributions of different isolating stages has been suggested to be closely associated with phylogenetic relatedness. Few studies have focused on the relative contributions of pre- versus post-zygotic mechanisms, and even fewer have been conducted under strict phylogenetic frameworks. Pre- and post-zygotic reproductive isolation stages have been investigated in the sister species Mussaenda kwangtungensis and M. pubescens var. alba. The two species have partly overlapping distribution ranges and flowering times, while the principal pollinators differed strikingly for them, demonstrating strong pre-zygotic isolations. Natural hybrids were detected by simple sequence repeat markers and their maternal parents were identified based on chloroplast gene sequences. Five out of 81 individuals were suggested to be hybrids that fall into the categories F2, BC1, and BC2 by the NewHybrids analysis. Interspecific crossings resulted in significantly reduced fruit set and seed germination rates. Phylogenetic analysis revealed short Kimura-2-parameter distance between M. kwangtungensis and M. pubescens var. alba. These findings strongly supported the hypothesis that for species with a closer phylogenetic relationship, pre-zygotic isolation plays an important part in limiting gene exchange in sympatric areas.

Experimental Evidence for Reproductive Isolation betweenAstrophytum coahuilense(H. Möller) Kayser andA. Capricornevar.Senile(Fric) Okumura (Cactaceae)

Abstract: Astrophytum coahuilense and Astrophytum capricorne var. senile have partly sympatric distributions in southern Coahuila and adjacent Zacatecas, Mexico. The apparent absence of natural hybrids in areas of sympatry has been noted by several authors, leading to speculation concerning the mechanisms of reproductive isolation. Experimental crosses between the two species revealed prezygotic isolating mechanisms in some cases and postzygotic mechanisms in others. Several different results were obtained from the cross-pollinations: 1) fruit was not produced and the ovules in the ovary of the floral remnant showed no development; 2) fruit contained incompletely developed, mostly non-germinant seeds; 3) apparently normal seed was produced, but many of the hybrid seedlings manifested morphological anomalies and mortality was high (>97%) due to chlorophyll deficiency. A series of experimental crosses between A. coahuilense and several allopatric taxa of the A. capricorne group revealed only postzygotic iso...

Recurrent epimutation of SDHC in gastrointestinal stromal tumors.

Succinate dehydrogenase (SDH) is a conserved effector of cellular metabolism and energy production, and loss of SDH function is a driver mechanism in several cancers. SDH-deficient gastrointestinal stromal tumors (dSDH GISTs) collectively manifest similar phenotypes, including hypermethylated epigenomic signatures, tendency to occur in pediatric patients, and lack of KIT/PDGFRA mutations. dSDH GISTs often harbor deleterious mutations in SDH subunit genes (SDHA, SDHB, SDHC, and SDHD, termed SDHx), but some are SDHx wild type (WT). To further elucidate mechanisms of SDH deactivation in SDHx-WT GIST, we performed targeted exome sequencing on 59 dSDH GISTs to identify 43 SDHx-mutant and 16 SDHx-WT cases. Genome-wide DNA methylation and expression profiling exposed SDHC promoter-specific CpG island hypermethylation and gene silencing in SDHx-WT dSDH GISTs [15 of 16 cases (94%)]. Six of 15 SDHC-epimutant GISTs occurred in the setting of the multitumor syndrome Carney triad. We observed neither SDHB promoter hypermethylation nor large deletions on chromosome 1q in any SDHx-WT cases. Deep genome sequencing of a 130-kbp (kilo-base pair) window around SDHC revealed no recognizable sequence anomalies in SDHC-epimutant tumors. More than 2000 benign and tumor reference tissues, including stem cells and malignancies with a hypermethylator epigenotype, exhibit solely a non-epimutant SDHC promoter. Mosaic constitutional SDHC promoter hypermethylation in blood and saliva from patients with SDHC-epimutant GIST implicates a postzygotic mechanism in the establishment and maintenance of SDHC epimutation. The discovery of SDHC epimutation provides a unifying explanation for the pathogenesis of dSDH GIST, whereby loss of SDH function stems from either SDHx mutation or SDHC epimutation.