Subgenome analysis of two southern hemisphere allotriploid species in Sphagnum (Sphagnaceae)

Sphagnum ×australe s.l. and S. ×falcatulum s.l. are both cryptic species complexes of gametophytically allodiploid and allotriploid cytotypes, with the allodiploid cytotype being one parent of the respective allotriploid cytotype. Phylogenetic analyses of S. ×australe s.l. and S. ×falcatulum s.l. were undertaken using sequences from two plastid loci and cloned sequences from three nuclear loci. Subgenomes from three subgenera were detected in allotriploid S. ×australe. The haploid parent of allotriploid S. ×australe was S. fimbriatum (subg. Acutifolia). One of the two subgenomes contributed by allodiploid S. australe was associated with S. strictum (subg. Rigida); the parental species associated with the other subgenome could not be identified, either at the species level or in terms of subgenus association. It may represent an early diverging lineage of subg. Sphagnum or a previously undetected subgenus. Allotriploid S. ×falcatulum has subgenomes from two subgenera. This study confirms prior findings that S. cuspidatum (subg. Cuspidata) was the haploid parent of allotriploid S. ×falcatulum. One of the two subgenomes contributed by allodiploid S. ×falcatulum was associated with subg. Cuspidata (species unidentified) and the second was associated with subg. Subsecunda (species unidentified). Recurrent allopolyploidy appears to have been associated with the complex evolutionary processes (inter-subgeneric hybridization, interploidal hybridization, double allopolyploidy) which resulted in each of these two allotriploids. The highest level of divergence detected among the subgenomes in each of these Sphagnum allotriploids is comparable to the average genetic divergence reported for angiosperm allopolyploids.     

Genetic evidence for hybridization between the native Spartina maritima and the introduced Spartina alterniflora (Poaceae) in South-West France: Spartina × neyrautii re-examined

 Spartina alterniflora, a perennial grass native to the North American Atlantic coast, was introduced during the 19th century in western Europe (Southern England and western France) where it hybridized with the native Spartina maritima. In England, the sterile hybrid S. × townsendii gave rise by chromosome doubling to the highly fertile allopolyploid Spartina anglica, which has now invaded many salt marshes and estuaries in western Europe, and has been introduced in several continents. In South-West France, another sterile hybrid was discovered in 1892 in the Bidassoa Estuary, and named Spartina × neyrautii. According to their morphology, some authors suggested that S. × neyrautii and S. × townsendii result from reciprocal crosses. During the 20th century, the hybridization site was severely disturbed, and surviving of S. × neyrautii was questioned. In this paper, various Spartina populations are investigated in the Basque region (France and Spain), and compared to the hybrid taxa formed in England (S. × townsendii and S. anglica). The samples were analyzed using molecular fingerprinting (RAPD and ISSR) and Chloroplast DNA sequence (trnL-trnT spacer, trnL intron and trnL-trnF spacer). In the Bidassoa estuary, a hybrid isolated clone has been found, that displays additive species-specific nuclear markers of S. maritima and S. alterniflora, and that is subsequently considered as a surviving clone of S. × neyrautii. The molecular analyses indicate that S. × neyrautii and S. × townsendii share the same maternal (S. alterniflora), and paternal (S. maritima) parental species, but also that the two independent hybridization events have involved different parental (nuclear) genotypes in England and in South-West France. Received July 12, 2002; accepted October 4, 2002 Published online: March 20, 2003     

The discovery of Antarctic RNA viruses: a new game changer

Antarctic ecosystems are dominated by micro‐organisms, and viruses play particularly important roles in the food webs. Since the first report in 2009 (López‐Bueno et al. 2009 ), ‘omic’‐based studies have greatly enlightened our understanding of Antarctic aquatic microbial diversity and ecosystem function (Wilkins et al. 2013 ; Cavicchioli 2015 ). This has included the discovery of many new eukaryotic viruses (López‐Bueno et al. 2009 ), virophage predators of algal viruses (Yau et al. 2011 ), bacteria with resistance to phage (Lauro et al. 2011 ) and mechanisms of haloarchaeal evasion, defence and adaptation to viruses (Tschitschko et al. 2015 ). In this issue of Molecular Ecology, López‐Bueno et al. ( 2015 ) report the first discovery of RNA viruses from an Antarctic aquatic environment. High sequence coverage enabled genome variation to be assessed for four positive‐sense single‐stranded RNA viruses from the order Picornavirales. By examining the populations present in the water column and in the lake's catchment area, populations of ‘quasispecies’ were able to be linked to local environmental factors. In view of the importance of viruses in Antarctic ecosystems but lack of data describing them, this study represents a significant advance in the field.     

Post-zygotic gametic selection due to endosperm balance number explains unusual chromosome numbers of 3×× 2× progeny in Solanum

 Crosses between triploid and diploid genotypes are usually the best sources of trisomics in potato as well as in several other crop species. However, 3×× 2× crosses between triploid (2n=3×=36; 2EBN) Solanum commersonii-S. tuberosum hybrids and diploid (2n= 2×=24; 2EBN) genotypes gave progenies with a high number of extra chromosomes, 29–36, suggesting that only eggs with 17–24 chromosomes produced embryos that reached full development. Our hypothesis is that although triploids produce eggs with a range of chromosome numbers, 3×× 2× crosses involving a 2×(2EBN) parent favor eggs with a high chromosome number. These eggs have higher probabilities of possessing the same endosperm balance number (EBN) value (i.e. 1) of gametes produced by the 2EBN diploid parent to give the required 2:1 maternal to paternal EBN ratio in the hybrid endosperm. Under this model, trisomics are produced only if the diploid parent has an EBN of 1. Based on our results and those reported in the literature, it is proposed that in 3×(2EBN) × 2×(2EBN) crosses the endosperm balance number exercises negative selection for gametes with a low chromosome number, and a corresponding low EBN, and positive selection for gametes with a high chromosome number and EBN. Received: 2 April 1998 / Revision accepted: 27 October 1998     

THE POPULATION GENETICS OF SPOROPHYTIC SELF‐INCOMPATIBILITY IN THREE HYBRIDIZING SENECIO (ASTERACEAE) SPECIES WITH CONTRASTING POPULATION HISTORIES

Hybridization generates evolutionary novelty and spreads adaptive variation. By promoting outcrossing, plant self‐incompatibility (SI) systems also favor interspecific hybridization because the S locus is under strong negative frequency‐dependent balancing selection. This study investigates the SI mating systems of three hybridizing Senecio species with contrasting population histories. Senecio aethnensis and S. chrysanthemifolius native to Sicily, form a hybrid zone at intermediate altitudes on Mount Etna, and their neo‐homoploid hybrid species, S. squalidus, has colonized disturbed urban habitats in the UK during the last 150 years. We show that all three species express sporophytic SI (SSI), where pollen incompatibility is controlled by the diploid parental genome, and that SSI is inherited and functions normally in hybrids. Large‐scale crossing studies of wild sampled populations allowed direct comparison of SSI between species and found that the main impacts of colonization in S. squalidus compared to Sicilian Senecio was a reduced number of S alleles, increased S allele frequencies, and increased interpopulation S allele sharing. In general, many S alleles were shared between species and the S locus showed reduced intra‐ and interspecific population genetic structure compared to molecular genetic markers, indicative of enhanced effective gene flow due to balancing selection.     

Vertebral deformities in interspecific diploid and triploid salmonid hybrids

Vertebral deformities in salmonid interspecific hybrids, some of which were triploidised, were assessed across three separate year classes during the freshwater life stage. Initially, eggs from a farmed Atlantic salmon Salmo salar were crossed with the sperm from a S. salar, arctic char Salvelinus alpinus or brown trout Salmo trutta. For S. salar × S. trutta, half the eggs were triploidised. In a second- and third-year class, the eggs from a farmed S. salar were crossed with the sperm from either a S. salar or a S. trutta, and half of each group was triploidised. In the two initial-year classes, all hybrids were larger than the S. salar controls, and triploid S. salar × S. trutta were larger than diploid counterparts. In the third-year class, the S. salar × S. trutta were smaller than the S. salar, in contrast to the initial 2 year classes, although the triploid hybrids were still larger than the diploids. In the third-year class, a high degree of spontaneous triploidy was also observed in the putative diploid groups (between 16 and 39%). Vertebral deformities were consistently higher in pressure-shocked triploids than diploids, irrespective of hybridisation, but there was no consistent effect of hybridisation among experiments. Although this study was not able to explain the contrasting results for vertebral deformities between year classes, triploid S. salar × S. trutta can demonstrate impressive freshwater growth that could be of interest for future farming programmes.     

Development of somatic hybrids Solanum × michoacanum Bitter. (Rydb.) (+) S. tuberosum L. and autofused 4x S. × michoacanum plants as potential sources of late blight resistance for potato breeding

Key message

Phytophthora infestans resistant somatic hybrids of S. × michoacanum (+) S. tuberosum and autofused 4 x S. × michoacanum were obtained. Our material is promising to introgress resistance from S. × michoacanum into cultivated potato background.

Abstract

Solanum × michoacanum (Bitter.) Rydb. (mch) is a wild diploid (2n = 2x = 24) potato species derived from spontaneous cross of S. bulbocastanum and S. pinnatisectum. This hybrid is a 1 EBN (endosperm balance number) species and can cross effectively only with other 1 EBN species. Plants of mch are resistant to Phytophthora infestans (Mont) de Bary. To introgress late blight resistance genes from mch into S. tuberosum (tbr), genepool somatic hybridization between mch and susceptible diploid potato clones (2n = 2x = 24) or potato cultivar Rywal (2n = 4x = 48) was performed. In total 18,775 calli were obtained from postfusion products from which 1,482 formed shoots. The Simple Sequence Repeat (SSR), Cleaved Amplified Polymorphic Sequences (CAPS) and Random Amplified Polymorphic DNA (RAPD) analyses confirmed hybrid nature of 228 plants and 116 autofused 4x mch. After evaluation of morphological features, flowering, pollen stainability, tuberization and ploidy level, 118 somatic hybrids and 116 autofused 4x mch were tested for late blight resistance using the detached leaf assay. After two seasons of testing three somatic hybrids and 109 4x mch were resistant. Resistant forms have adequate pollen stainability for use in crossing programme and are a promising material useful for introgression resistance from mch into the cultivated potato background.     

Molecular fingerprinting of peppermint (Mentha piperita) and some Mentha hybrids by sequencing and RFLP analysis of the 5S rRNA Non-Transcribed Spacer (NTS) region

Hybridization of species belonging to the genus Mentha is quite common. However, the indicators of hybridity are many and make Mentha hybrids' identification difficult. By using the same molecular strategy that allowed us to unequivocally identify some Mentha species, we amplified the Not-Transcribed-Spacer (NTS) of the 5S-rRNA gene to characterize the industrial crop peppermint, M. × piperita and some important Mentha interspecific hybrids: M. × dalmatica, M. × dumetorum, M. × rotundifolia, M. × maximilianea, M. × smithiana, M. × verticillata, M. × villosa. DNA amplification, sequence and cluster analysis revealed differences in the 5S-rRNA NTS region of Mentha hybrids. Peppermint and all other hybrids were unequivocally discriminated by RFLP analysis by using TaqI restriction enzyme, while a further discrimination between M. × dumetorum and M. × verticillata was obtained by XhoI restriction enzyme. Essential oil composition showed clustering patterns similar to DNA fingerprint, with a clear discrimination between plants producing menthofuran (e.g. M. aquatica and its related hybrids, including peppermint) and those containing piperitenone oxide (M. longifolia and its related hybrids).     

Hybridization of Senecio squalidus and S. viscosus and Introgression of Genes from Diploid into Tetraploid Senecio Species

Natural triploid hybrids (Senecio x londinensis Lousley) betweenS. squalidus L. (2n = 20) and 5. viscosus L. (2n = 40) are fairlyfrequently found in Britain. Under glasshouse conditions bothnatural and artificial hybrids displayed very low levels ofseed fertility and gave rise to morphologically diverse F₂ plantsat about the triploid or pentaploid chromosome levels. By theF₄ generation, progeny of a F₂ pentaploid plant had somaticchromosome numbers near to the tetraploid level and considerablyincreased pollen and seed fertilities. Such fertile tetraploidsegregants of S. x londinensis permit the introgression of S.squalidus genes into S. viscosus, and may indicate the courseof introgression into other tetraploid species of Senecio. Senecio, hybridization, introgression     

The arctic-alpine polyploids Cerastium alpinum and C. nigrescens (Caryophyllaceae) in a sympatric situation: breakdown of species integrity?

 The high polyploids Cerastium alpinum (8x) and C. nigrescens (12x) were investigated in a mixed population in central Norway to evaluate whether hybridisation has resulted in continuous variation in morphology and genetic markers, a hypothesis previously proposed to account for the extensive taxonomic confusion in this species group. Isozyme, fertility, and morphological (37 characters) variation were examined among 347, 265, and 237 plants, respectively. A PCO analysis based on 23 quantitative morphological characters identified two main groups, corresponding to C. alpinum and C. nigrescens. The groups were also clearly separated in isozyme markers, several qualitative morphological characters, and chromosome numbers. Only 20 plants (8.4%) were more or less intermediate in the PCO analysis. These plants had a parental or hybrid isozyme multilocus phenotype and typically few well-developed anthers, low pollen stainability, and no seed set. Several plants within the C. nigrescens group also had reduced pollen stainability. These results indicate that later-generation hybrids and/or backcrosses towards C. nigrescens are formed. Thus, interspecific, interploidal gene flow probably occurs, but at rates that are insufficient to break down species integrity. Received July 5, 2000 Accepted August 2, 2001     

Yield performance of 14 novel inter- and intra-species Miscanthus hybrids across Europe

Miscanthus, a C4 perennial rhizomatous grass from Asia is a leading candidate for the supply of sustainable biomass needed to grow the bioeconomy. European Miscanthus breeding programmes have recently produced a new range of seeded hybrids with the objective of increasing scalability to large acreages limited by current clonal propagation. For the EU-GRACE project, new replicated field trials were established in seven locations across Europe in 2018 with eight intraspecific M. sinensis hybrids (sin × sin) and six M. sacchariflorus × M. sinensis (sac × sin) from Dutch and UK breeding programmes, respectively, with clonal Miscanthus × giganteus. The planting density of the sin × sin was double that of sac × sin (30,000 & 15,000 plants ha⁻¹), creating commercially relevant upscaling comparisons between systems. Over the first 3 years, the establishment depended on location and hybrid. The mature sin × sin hybrids formed tight tufts of shoots up to 2.5 m tall which flower and senesce earlier than the taller sac × sin hybrids. Following the third growing season, the highest yields were recorded in Northern Italy at a low altitude (average 13.7 (max 21) Mg DM ha⁻¹) and the lowest yielding was on the industrially damaged marginal land site in Northern France (average 7.0 (max 10) Mg DM ha⁻¹). Moisture contents at spring harvest were lowest in Croatia (21.7%) and highest in Wales, UK (41.6%). Overall, lower moisture contents at harvest, which are highly desirable for transport, storage and for most end-use applications, were found in sin × sin hybrids than sac × sin (30% and 40%, respectively). Yield depended on climate interactions with the hybrid and their associated planting systems. The sin × sin hybrids appeared better adapted to northern Europe and sac × sin hybrids to southern Europe. Longer-term yield observations over crop lifespans will be needed to explore the biological (yield persistence) and economic costs and benefits of the different hybrid systems.     

Natural hybridization between <Emphasis Type="Italic">Phlomis lycia</Emphasis> D. Don × <Emphasis Type="Italic">P. bourgaei</Emphasis> Boiss., (Lamiaceae) revealed by RAPD markers

Randomly Amplified Polymorphic DNA markers (RAPD) were used to assess the hybrid identity of individuals sampled as Phlomis × termessi Davis. Out of 95 primers screened, 11 primers produced reproducible amplification patterns used for discrimination of P. × termessi and their parents. Eleven primers produced 81 bands. Forty two percent of the RAPD bands existed in parents. Of the 54 bands found in P. lycia, 19 were found only in this species and 7 of these were monomorphic. Similarly, of 57 RAPD bands observed in P. bourgaei, 18 were found only in P. bourgaei and 6 of these were monomorphic. Among hybrid individuals, 35 of the 73 markers were monomorphic. Fifteen of these existed in individual parents showing that parents were homozygous for these markers. Of the 35 monomorphic bands observed among hybrid individuals, 5 were present in the samples of one of the parents and completely absent from the samples of the other; therefore, additive inheritance is indicated. Of the 5 additive bands, 1 was inherited from P. bourgaei and 4 were inherited from P. lycia. Among 38 polymorhic markers observed in hybrid individuals, 9 were new and hybrid-specific. Pollen fertility was also investigated. Mean pollen fertility for P. lycia and P. bourgaei was 93% and 97% respectively. However, mean pollen fertility for hybrids was 65% (±10.5).     

A farnesyl pyrophosphate synthase gene expressed in pollen functions in S‐RNase‐independent unilateral incompatibility

Multiple independent and overlapping pollen rejection pathways contribute to unilateral interspecific incompatibility (UI). In crosses between tomato species, pollen rejection usually occurs when the female parent is self‐incompatible (SI) and the male parent self‐compatible (SC) (the ‘SI × SC rule’). Additional, as yet unknown, UI mechanisms are independent of self‐incompatibility and contribute to UI between SC species or populations. We identified a major quantitative trait locus on chromosome 10 (ui10.1) which affects pollen‐side UI responses in crosses between cultivated tomato, Solanum lycopersicum, and Solanum pennelliiLA0716, both of which are SC and lack S‐RNase, the pistil determinant of S‐specificity in Solanaceae. Here we show that ui10.1 is a farnesyl pyrophosphate synthase gene (FPS2) expressed in pollen. Expression is about 18‐fold higher in pollen of S. pennellii than in S. lycopersicum. Pollen with the hypomorphic S. lycopersicum allele is selectively eliminated on pistils of the F₁ hybrid, leading to transmission ratio distortion in the F₂ progeny. CRISPR/Cas9‐generated knockout mutants (fps2) in S. pennelliiLA0716 are self‐sterile due to pollen rejection, but mutant pollen is fully functional on pistils of S. lycopersicum. F₂ progeny of S. lycopersicum × S. pennellii (fps2) show reversed transmission ratio distortion due to selective elimination of pollen bearing the knockout allele. Overexpression of FPS2 in S. lycopersicum pollen rescues the pollen elimination phenotype. FPS2‐based pollen selectivity does not involve S‐RNase and has not been previously linked to UI. Our results point to an entirely new mechanism of interspecific pollen rejection in plants.     

Molecules and morphology reveal a new aberrant harvestman genus of Ortholasmatinae (Opiliones,Dypsnoi, Nemastomatidae) from Mexico

Ortholasmatinae is one of the most striking groups of Opiliones, because its members exhibit a peculiar dorsal ornamentation pattern of cells, formed by fusion of anvil-shaped tubercles. During the systematic revision of several specimens of ortholasmatines, we found specimens with shared morphological characters with two genera Trilasma Goodnight & Goodnight, 1942 and Dendrolasma Banks, 1894. In order to investigate their phylogenetic relationships, we performed model-based phylogenetic analyses using the mitochondrial protein-encoding cytochrome c oxidase subunit 1 (CO1) and the complete nuclear ribosomal small subunit 18S, including members of the five genera of this subfamily. The results support the creation of Cryptolasma Cruz-López, Cruz-Bonilla & Francke, gen. nov., and two new species: Cryptolasma aberrante Cruz-López, Cruz-Bonilla & Francke, sp. nov. (type species) and Cryptolasma citlaltepetl Cruz-López, Cruz-Bonilla & Francke, sp. nov., which are from Xalapa and Orizaba Volcano, Veracruz, respectively. Additionally, our results suggest Ruaxphilos petrukenvitchou Goodnight & Goodnight, 1945 is a nomina dubia. This study represents the first molecular hypothesis of Ortholasmatinae relationships, with Cryptolasma as the sister of the remaining ortholasmatine.www.zoobank.org/urn:lsid:zoobank.org:pub:3730EE48-9802-4B06-9DDF-EC1AFE9A3B61     

Hybridisation between Brassica napus L. and Raphanus raphanistrum L. under agronomic field conditions

The frequency of hybridisation between Brassica napus L. and Raphanus raphanistrum L. under agronomic conditions was assessed in field experiments, where R. raphanistrum were randomly planted at two different densities into large plots of B. napus. An acetolacate synthase (ALS)-inhibiting herbicide-resistant trait was used to detect potential hybrid individuals. No hybrids were detected amongst 25,000 seedlings grown from seed collected from R. raphanistrum plants. Two hybrids were obtained from more than 52-million B. napus seedlings. Both hybrids were characterised as amphidiploids (AACCRrRr, 2n = 56) and were fertile. The frequency of hybridisation into B. napus in this experiment using male-fertile B. napus was 4 × 10^–8. Received: 31 August 2000 / Accepted: 23 January 2001     

Natural hybridization in JapaneseCalamagrostis

Morphological and pollen studies were made for the collections belonging toCalamagrostis langsdorffii, C. Sachalinensis, C. longiseta, C. nana and various intermediates between these species from central Honshu, the chromosome numbers of which were reported in a previous paper (Tateoka, 1976). Additional collections made from the Akaishi Range in 1976 were also subjected to the present work. The intermediates showed 0%, or nearly 0%, pollen stainability and a mosaic of morphological features of putative parental taxa which can be estimated to occur for F₁ hybrids. This estimation was sustained by a comparison with the pattern of character expressions for the artificially raised F₁ hybrids of various European species ofCalamagrostis as reported by Nygren (1962). These results as well as field observations strongly suggested that the intermediates may be wholly or almost wholly F₁ hybrids. The hybrids were sometimes found in abundance within a limited area, but seemed to have little capacity to migrate from the places where they originated. The following combinations, which were not previously recorded, were disclosed:C. longiseta×C. sachalinensis, C. langsdorffii×C. nana, C. langsdorffii×C. longiseta, C. longiseta×C. nana. Possibilities of introgression in these hybridizing species were discussed. The hybrids were tetraploids except one hexaploid which was discovered in a mixed population ofC. nana, C. longiseta and their putative F₁ hybrids at the tetraploid level. The hexaploid in question, which seemed to have resulted from the participation of an unreduced gamete, was very similar morphologically toC. nana subsp.hayachinensis distributed in the mountain far distant from central Honshu. Geographical distributions of the hybrid plants were surveyed through examination of the herbarium specimens.     

Characterisation,applicability and defect analysis for tiles nanostructure of quantum dot cellular automata

Quantum dot cellular automata (QCA) is an emerging technology for nanoscale computing. In this paper, our contribution is to enhance the applicability of 3 × 3 tile nanostructure in QCA. We introduced few novel 3 × 3 active-type tiles, namely triple fan-output tiles or butterfly tile, five-input majority voter (MV) and fault-tolerant 3 × 3 tile-coupled majority–minority voter. We also explored the cascading effect on 3 × 3 tile. Triple fan-out tile is applicable to drive three outputs simultaneously. Five-input MV, which is another active-type tile we proposed, is applicable for designing five-variable or five addressing, advanced computations. In passive-type 3 × 3 tiles, we introduce symmetrical baseline tiles and asymmetrical fully populated fan-in tiles. QCA design was found to be having high error during fabrication of device. We made exhaustive simulation for analysing the defect that can occur during fabrication process. Defects such as extra cell deposition, missing cell deposition and displaced cell deposition for QCA are prone to occur. Fault analyses of the above-mentioned 3 × 3 tiles are performed. We also focused on the effect of extra dot or missing dot deposition, during the chemical synthesis phase.     

Comparison of pollen grain morphological features of selected species of the genus Crataegus (Rosaceae) and their spontaneous hybrids

The aim of our study was to verify, on the basis of statistical analyses of nine quantitative morphological features of pollen grains, the hypothesis that pollen grains of three parental species of Crataegus (C. laevigata, C. monogyna, C. rhipidophylla) differed from the pollen of three spontaneous hybrids of these species (C. × macrocarpa, C. × media, C. × subsphaericea). Contrast analysis revealed that a majority of the pollen features of hybrid species were characterized by significantly higher mean values than those of parental species. Analysis of pollen shape classes indicated that the parental species clearly differed from each other in contrast with hybrids, which were characterized by a similar proportion of pollen in individual pollen shape classes. Statistical analyses showed that the pollen grains of two parental species, C. laevigata and C. monogyna, were most similar to one another. Pollen grains of typical C. rhipidophylla were similar to the pollen of hybrids and the mean values of almost all studied pollen features [P, E, Exp, Exp/P, Le, d, d/E (PAI)] of C. rhipidophylla var. rhipidophylla were intermediate between those of C. monogyna and C. rhipidophylla var. lindmanii. This corroborates Zieliński's conjecture that C. rhipidophylla is probably an old, conserved hybrid between C. monogyna and C. calycina (= C. lindmanii = C. rhipidophylla var. lindmanii). According to the analysis of canonical variables, C. × macrocarpa and C. × media pollen grains were most similar. C. × subsphaericea and C. rhipidophylla var. rhipidophylla and C. rhipidophylla var. lindmanii formed another pair (group), and C. monogyna and C. laevigata constituted separate ‘single species groups’. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 172 , 555–571.     

<Emphasis Type="Italic">Hieracium</Emphasis> subgen. <Emphasis Type="Italic">Pilosella</Emphasis>: pollen stainability in sexual,apomictic and sterile plants

Two pollen stainability tests (Alexander’s stain and acetocarmine) were used to detect differences in pollen viability of the sexual, apomictic and sterile plants of Hieracium subgen. Pilosella. In sexual taxa (Hieracium bauhini and H. densiflorum), the average stainability was 93.7–98.4%. Similarly high stainability (92.2–97.2%) was found in the apomictic Hieracium pilosellinum and in the majority of the apomictic populations (or plants) of the pentaploid and hexaploid H. bauhini. In some apomictic plants of Hieracium bauhini the average pollen stainability was 49.0–75.4%. The lowest pollen stainability was found in the sterile plants, i.e. the triploid H. pistoriense (33.6%) and the pentaploid H. brachiatum (29.6%).     

Effect of genome-plastome interaction on meiosis and pollen development in Oenothera species and hybrids

 Oenothera villaricae Dietrich and Oe. picensis Dietrich, complete translocation heterozygotes, are fully interfertile, giving rise to six discrete classes of true-breeding hybrids from a reciprocal cross. Associated with each parent and hybrid is a characteristic abortive non-staining pollen fraction easily distinguished from fully developed pollen under the light microscope. Pollen abortion has been associated with translocation rings in other angiosperm species, and may characterize such systems. The abortive pollen fraction is significantly different between reciprocal Oenothera hybrids, however (P<0.001), indicative of partial cytoplasmic control. Pollen abortion is most severe in the F₁ hybrid generation, and ameliorates with successive generations of hybrid self-fertilization. Three-way analysis of variance shows significant effects on pollen stainability (a measure of the non-abortive fraction) for nucleus, cytoplasm and selfed hybrid generation, individually or in combinations. This result suggests a combined nucleocytoplasmic basis for the pollen abortion. Correlated with the observation of increased pollen abortion in Oenothera hybrids are meiotic findings of broken chromosome rings (chains, univalents), asymmetric anaphase chromosome distributions and trinucleate tetrads. To test the hypothesis that such anomalous meiotic events play a role in the mechanism of pollen abortion, meiotic disjunction frequency was determined for each parent, F₁ and F₉ selfed hybrid accessions. Three-way analysis of variance shows levels of significance comparable to those noted for pollen stainability (P<0.001) for effects of nucleus, cytoplasm and selfed hybrid generation on disjunction frequency. A high degree of correlation (r ²=0.984) is noted between disjunction frequency and pollen stainability. We conclude that the abortive pollen grains are indeed the products of nondisjunctional meiotic events, which themselves are consequences of hybrid nucleocytoplasmic incompatibility. Received: 28 May 1997 / Revision accepted: 23 July 1997