Callose in sporogenesis: Novel composition of the inner spore wall in hornworts
Sporogenesis is a developmental process that defines embryophytes and involves callose, especially in the production of the highly protective and recalcitrant spore/pollen wall. Until now, hornworts, leptosporangiate ferns and homosporous lycophytes are the only major plant groups in which the involvement of callose in spore development is equivocal. Through aniline blue fluorescence and immunogold labeling in the transmission electron microscope, we provide indisputable evidence for the presence of callose in the spore wall of five hornwort genera, but not in the derived , an epiphyte that produces multicellular spores. We present evidence that callose appears in the developing spore wall and is retained throughout development as a wall constituent of the intine or inner spore wall, a novel location for this polysaccharide in embryophytes. In endosporic and multicellular spores/pollen of , the liverwort , and , callose appears in the newly formed cell walls only following the first mitotic division. Further probing for other wall polymers in hornworts reveals the presence of cellulose (Calcofluor fluorescence) in the spore intine, aperture and around the equatorial girdle. Further immunogold labeling with monoclonal antibodies identifies pectin and hemicellulose in hornwort intines. The persistence of callose, a typically transient cell wall constituent, with cellulose, pectins and hemicellulose in the intine, supports specialized functions of callose in spores of hornworts that include reduced water loss when spores are dry and mechanical flexibility to withstand desiccation.
Chromosome numbers in Tripleurospermum Sch. Bip. (Asteraceae) and closely related genera: relationships between ploidy level and stomatal length
This study includes 24 reports of chromosome counts in Tripleurospermum Sch. Bip. (20 taxa), Anthemis L. (1 species) and Matricaria L. (3 taxa) belonging to the tribe Anthemideae of the family Asteraceae from Turkey. Chromosome numbers of these taxa are 2n = 2x = 18, 3x = 27 and 4x = 36. Nine counts are new reports, one is not consistent with previous report, and the remainder confirm earlier information. Statistically significant differences depending on ploidy level, stomatal length or environmental factors such as altitude were determined in Tripleurospermum. Several systematic and evolutionary aspects of the genera are discussed in the light of chromosomal data.
Malagasy Dracaena Vand. ex L. (Ruscaceae): an investigation of discrepancies between morphological features and spatial genetic structure at a small evolutionary scale
Malagasy Dracaena (Ruscaceae) are divided into four species and 14 varieties, all of them showing a high level of morphological diversity and a putatively artefactual circumscription. In order to reveal relationships between those entangled entities, a span of Malagasy Dracaena were sampled and analyzed using cpDNA sequences and AFLP. The cpDNA analyses resolved three biogeographic clades that are mostly inconsistent with morphology, since similar phenotypes are found across the three clades. Bayesian inference clustering analyses based on the AFLP were not in accordance with the cpDNA analysis. This result might be explained by (1) a recent origin of the Malagasy species of Dracaena with an incomplete sorting of chloroplast lineages; (2) a high amount of hybridizations; (3) a complex migration pattern. Interestingly, when the AFLP are analyzed using the parsimony criterion, a trend towards a directional evolution of inflorescence types and ecological features was observed. This might be considered either as phenotypic plasticity and/or as the result of fast evolution in flower characters according to habitat preferences. Overall, our results point to the difficulty of defining evolutionarily significant units in Malagasy Dracaena, emphasizing the complex speciation processes taking place in tropical regions.
Fagaceae pollen from the early Cenozoic of West Greenland: revisiting Engler's and Chaney's Arcto-Tertiary hypotheses
In this paper we document Fagaceae pollen from the Eocene of western Greenland. The pollen record suggests a remarkable diversity of the family in the early Cenozoic of Greenland. Extinct Fagaceae pollen types include , which extends at least back to the Paleocene, and two ancestral pollen types with affinities to the Eurasian Group Ilex and the western North American Group Protobalanus. In addition, modern lineages of Fagaceae are unambiguously represented by pollen of Group Lobatae/Quercus, and three Castaneoideae pollen types. These findings corroborate earlier findings from Axel Heiberg Island that Fagaceae were a dominant element at high latitudes during the early Cenozoic. Comparison with coeval or older mid-latitude records of modern lineages of Fagaceae shows that modern lineages found in western Greenland and Axel Heiberg likely originated at lower latitudes. Further examples comprise (possibly) , , , , and others. Thus, before fossils belonging to modern northern temperate lineages will have been recovered from older (early Eocene, Paleocene) strata from high latitudes, Engler's hypothesis of an Arctic origin of the modern temperate woody flora of Eurasia, termed 'Arcto-Tertiary Element', and later modification by R. W. Chaney and H. D. Mai ('Arcto-Tertiary Geoflora') needs to be modified.
Genome-wide repeat dynamics reflect phylogenetic distance in closely related allotetraploid (Solanaceae)
sect. is a group of four allotetraploid species originating from a single allopolyploidisation event approximately 5 million years ago. Previous phylogenetic analyses support the hypothesis of as sister to the other three species. This is concordant with changes in genome size, separating those with genome downsizing () from those with genome upsizing (, , ). However, a recent analysis reflecting genome dynamics of different transposable element families reconstructed greater similarity between and the Revillagigedo Island taxa ( and ), thereby placing as sister to the rest of the group. This could reflect a different phylogenetic hypothesis or the unique evolutionary history of these particular elements. Here we re-examine relationships in this group and investigate genome-wide patterns in repetitive DNA, utilising high-throughput sequencing and a genome skimming approach. Repetitive DNA clusters provide support for as sister to the rest of the section, with sister to the two Revillagigedo Island species. Clade-specific patterns in the occurrence and abundance of particular repeats confirm the original ( ( ( + ))) hypothesis. Furthermore, overall repeat dynamics in the island species and confirm their similarity to and the distinctive patterns between these three species and . Together these results suggest that broad-scale repeat dynamics do in fact reflect evolutionary history and could be predicted based on phylogenetic distance.
Pathways to polyploidy: indications of a female triploid bridge in the alpine species (Ranunculaceae)
Polyploidy is one of the most important evolutionary processes in plants. In natural populations, polyploids usually emerge from unreduced gametes which either fuse with reduced ones, resulting in triploid offspring (triploid bridge), or with other unreduced gametes, resulting in tetraploid embryos. The frequencies of these two pathways, and male versus female gamete contributions, however, are largely unexplored. occurs with diploid, triploid and autotetraploid cytotypes in the Alps, whereby diploids are mostly sexual, while tetraploids are facultative apomicts. To test for the occurrence of polyploidization events by triploid bridge, we investigated 551 plants of natural populations via flow cytometric seed screening. We assessed ploidy shifts in the embryo to reconstruct female versus male gamete contributions to polyploid embryo and/or endosperm formation. Seed formation via unreduced egg cells (B hybrids) occurred in all three cytotypes, while only in one case both gametes were unreduced. Polyploids further formed seeds with reduced, unfertilized egg cells (polyhaploids and aneuploids). Pollen was highly variable in diameter, but only pollen >27 μm was viable, whereby diploids produced higher proportions of well-developed pollen. Pollen size was not informative for the formation of unreduced pollen. These results suggest that a female triploid bridge via unreduced egg cells is the major pathway toward polyploidization in , maybe as a consequence of constraints of endosperm development. Triploids resulting from unreduced male gametes were not observed, which explains the lack of obligate sexual tetraploid individuals and populations. Unreduced egg cell formation in diploids represents the first step toward apomixis.
Phylogenomics of the pantropical Connaraceae: revised infrafamilial classification and the evolution of heterostyly
Connaraceae is a pantropical family of about 200 species containing lianas and small trees with remarkably diverse floral polymorphisms, including distyly, tristyly, homostyly, and dioecy. To date, relationships within the family have not been investigated using a targeted molecular phylogenetic treatment, severely limiting systematic understanding and reconstruction of trait evolution. Accordingly, their last infrafamilial classification was based only on morphological data. Here, we used phylogenomic data obtained using the Angiosperms353 nuclear target sequence capture probes, sampling all tribes and almost all genera, entirely from herbarium specimens, to revise infrafamilial classification and investigate the evolution of heterostyly. The backbone of the resulting molecular phylogenetic tree is almost entirely resolved. Connaraceae consists of two clades, one containing only the African genus (4 or 5 species), which we newly recognize at the subfamily level. Vegetative and reproductive synapomorphies are proposed for Manotoideae. Within Connaroideae, Connareae is expanded to include the former Jollydoreae. The backbone of Cnestideae, which contains more than half of the Connaraceae species, remains incompletely resolved. Reconstructions of reproductive system evolution are presented that tentatively support tristyly as the ancestral state for the family, with multiple parallel losses, in agreement with previous hypotheses, plus possible re-gains. However, the great diversity of stylar polymorphisms and their phylogenetic lability preclude a definitive answer. Overall, this study reinforces the usefulness of herbarium phylogenomics, and unlocks the reproductive diversity of Connaraceae as a model system for the evolution of complex biological phenomena.