A Review of the Developmental Processes and Selective Pressures Shaping Aperture Pattern in Angiosperms.

Pollen grains of flowering plants display a fascinating diversity of forms. The observed diversity is determined by the developmental mechanisms involved in the establishment of pollen morphological features. Pollen grains are generally surrounded by an extremely resistant wall displaying apertures that play a key role in reproduction, being the places at which pollen tube growth is initiated. Aperture number, structure, and position (collectively termed 'aperture pattern') are determined during microsporogenesis, which is the earliest step of pollen ontogeny. Here, we review current knowledge about aperture pattern developmental mechanisms and adaptive significance with respect to plant reproduction and how advances in these fields shed light on our understanding of aperture pattern evolution in angiosperms.

Phylogenetic analysis of fossil flowers using an angiosperm-wide data set: proof-of-concept and challenges ahead.

PREMISE: Significant paleobotanical discoveries in recent decades have considerably improved our understanding of the early evolution of angiosperms and their flowers. However, our ability to test the systematic placement of fossil flowers on the basis of phylogenetic analyses has remained limited, mainly due to the lack of an adequate, angiosperm-wide morphological data set for extant taxa. Earlier attempts to place fossil flowers phylogenetically were, therefore, forced to make prior qualitative assessments of the potential systematic position of fossils and to restrict phylogenetic analyses to selected angiosperm subgroups. METHODS: We conduct angiosperm-wide molecular backbone analyses of 10 fossil flower taxa selected from the Cretaceous record. Our analyses make use of a floral trait data set built within the framework of the eFLOWER initiative. We provide an updated version of this data set containing data for 28 floral and two pollen traits for 792 extant species representing 372 angiosperm families. RESULTS: We find that some fossils are placed congruently with earlier hypotheses while others are found in positions that had not been suggested previously. A few take up equivocal positions, including the stem branches of large clades. CONCLUSIONS: Our study provides an objective approach to test for the phylogenetic position of fossil flowers across angiosperms. Such analyses may provide a complementary tool for paleobotanical studies, allowing for a more comprehensive understanding of fossil phylogenetic relationships in angiosperms. Ongoing work focused on extending the sampling of extant taxa and the number of floral traits will further improve the applicability and accuracy of our approach.

Polygamy or subdioecy? The impact of diallelic self-incompatibility on the sexual system in Fraxinus excelsior (Oleaceae).

How flowering plants have recurrently evolved from hermaphroditism to separate sexes (dioecy) is a central question in evolutionary biology. Here, we investigate whether diallelic self-incompatibility (DSI) is associated with sexual specialization in the polygamous common ash (Fraxinus excelsior), which would ultimately facilitate the evolution towards dioecy. Using interspecific crosses, we provide evidence of strong relationships between the DSI system and sexual phenotype. The reproductive system in F. excelsior that was previously viewed as polygamy (co-occurrence of unisexuals and hermaphrodites with varying degrees of allocation to the male and female functions) and thus appears to actually behave as a subdioecious system. Hermaphrodites and females belong to one SI group and functionally reproduce as females, whereas males and male-biased hermaphrodites belong to the other SI group and are functionally males. Our results offer an alternative mechanism for the evolution of sexual specialization in flowering plants.

Effect of aperture number on pollen germination, survival and reproductive success in Arabidopsis thaliana.

Background and Aims: Pollen grains of flowering plants display a fascinating diversity of forms, including diverse patterns of apertures, the specialized areas on the pollen surface that commonly serve as the sites of pollen tube initiation and, therefore, might play a key role in reproduction. Although many aperture patterns exist in angiosperms, pollen with three apertures (triaperturate) constitutes the predominant pollen type found in eudicot species. The aim of this study was to explore whether having three apertures provides selective advantages over other aperture patterns in terms of pollen survival, germination and reproductive success, which could potentially explain the prevalence of triaperturate pollen among eudicots. Methods: The in vivo pollen germination, pollen tube growth, longevity and competitive ability to sire seeds were compared among pollen grains of Arabidopsis thaliana with different aperture numbers. For this, an arabidopsis pollen aperture series was used, which included the triaperturate wild type, as well as mutants without an aperture (inaperturate) and with more than three apertures. Key Results: Aperture number appears to influence pollen grain performance. In most germination and longevity experiments, the triaperturate and inaperturate pollen grains performed better than pollen with higher aperture numbers. In mixed pollinations, in which triaperturate and inaperturate pollen were forced to compete with each other, the triaperturate pollen outperformed the inaperturate pollen. Conclusions: Triaperturate pollen grains might provide the best trade-off among various pollen performance traits, thus explaining the prevalence of this morphological trait in the eudicot clade.

More than sixty origins of pantoporate pollen in angiosperms.

PREMISE OF THE STUDY: Apertures in pollen grains are key structures of the wall, involved in pollen tube germination and exchanges with the environment. Aperture types in angiosperms are diverse, but pollen with one and three apertures (including monosulcate and tricolpate, respectively) are the two most common types. Here, we investigate the phylogenetic distribution in angiosperms of pollen with many round, scattered apertures called pantoporate pollen. METHODS: We constructed a morphological data set with species producing pantoporate pollen and representative angiosperm species with other pollen types, sampled from every angiosperm order, with a total of 1260 species distributed in 330 families. This data set was analyzed with parsimony to characterize the phylogenetic distribution of pantoporate pollen in angiosperms. KEY RESULTS: We show that pantoporate pollen is distributed throughout most of the angiosperm tree, including early diverging angiosperms, monocots, and eudicots. However, this pollen type is usually restricted to a few species in a given group, and is seldom fixed at large taxonomical scales, with a few notable exceptions. CONCLUSIONS: Pantoporate pollen evolved many times during angiosperm history, but the persistence of this morphology in the long term is infrequent. This distribution pattern could indicate conflicting short-term and long-term selective pressures, pantoporate pollen being selected in the short run, but eliminated in the long run. Biological hypotheses supporting this scenario are discussed, in the context of both theoretical and empirical data on pollen biology.

Aperture number influences pollen survival in Arabidopsis mutants.

PREMISE OF THE STUDY: Pollen grains are subject to intense dehydration before dispersal. They rehydrate after landing on a stigma or when placed in humid environment by absorbing water from the stigma or surroundings. Resulting fluctuations in water content cause pollen grains to undergo significant changes in volume. Thus, morphological or structural adaptations might exist to help pollen adjust to sudden volume changes, though little is known about the correlation between pollen morphology and its ability to accommodate volume changes. We studied the effect of one morphological feature of pollen grains, the aperture number, on pollen wall resistance to water inflow in Arabidopsis thaliana. METHODS: We used three Arabidopsis thaliana mutants that differ in the number of apertures in their pollen (zero, four, or a mix of four to eight, respectively) and the wild type with pollen with three apertures. We tested pollen survival in solutions with various mannitol concentrations. KEY RESULTS: The number of intact pollen grains increased with increasing mannitol concentration for all pollen morphs tested. At a given mannitol concentration, however, an increase in aperture number was associated with an increase in pollen breakage. CONCLUSIONS: Aperture patterns, i.e., number, shape, and position, influence the capacity to accommodate volume variations in pollen grains. When subjected to water inflow, pollen grains with few apertures survive better than pollen with many apertures. Trade-offs between survival and germination are likely to be involved in the evolution of pollen morphology.

Evolutionary stasis in pollen morphogenesis due to natural selection.

The contribution of developmental constraints and selective forces to the determination of evolutionary patterns is an important and unsolved question. We test whether the long-term evolutionary stasis observed for pollen morphogenesis (microsporogenesis) in eudicots is due to developmental constraints or to selection on a morphological trait shaped by microsporogenesis: the equatorial aperture pattern. Most eudicots have three equatorial apertures but several taxa have independently lost the equatorial pattern and have microsporogenesis decoupled from aperture pattern determination. If selection on the equatorial pattern limits variation, we expect to see increased variation in microsporogenesis in the nonequatorial clades. Variation of microsporogenesis was studied using phylogenetic comparative analyses in 83 species dispersed throughout eudicots including species with and without equatorial apertures. The species that have lost the equatorial pattern have highly variable microsporogenesis at the intra-individual and inter-specific levels regardless of their pollen morphology, whereas microsporogenesis remains stable in species with the equatorial pattern. The observed burst of variation upon loss of equatorial apertures shows that there are no strong developmental constraints precluding variation in microsporogenesis, and that the stasis is likely to be due principally to selective pressure acting on pollen morphogenesis because of its implication in the determination of the equatorial aperture pattern.

Sex expression and reproductive biology in a tree species, Fraxinus excelsior L.

We investigated Fraxinus excelsior breeding system using field data collected in a natural population and in a seed orchard. First, we attested functional trioecy (co-occurrence of males, hermaphrodites and females), with males producing pollen, hermaphrodites producing both pollen and seeds simultaneously, and females producing seeds. Second, we found that the reproductive system of F. excelsior was not labile, as sex expression seemed to be stable through time. Third, gender is genetically determined since different trees belonging to the same clone in the orchard exhibit similar sexual phenotypes.

The Drosophilidae (Diptera) of the Scattered Islands, with the description of a novel association with Leptadenia madagascariensis Decne. (Apocynaceae).

Thirteen drosophilid species belonging to seven genera and two subfamilies are reported from three coral islands (namely Europa, Juan de Nova and Glorioso) that belong to the Scattered Islands in the Indian Ocean. Five species are cosmopolitan and five are African. Three are endemic to the insular Western Indian Ocean, including a presumably new Scaptodrosophila species. On the island of Juan de Nova, most captured flies had pollinia attached to the bases of their proboscis. DNA analysis using the rbcl gene revealed that these pollinia belong to the genus Leptadenia (Apocynaceae), of which a single species L. madagascariensis, endemic in Madagascar and Comoros, is present in this island. This is the first reported association between this plant and drosophilids.

Correlation between pollen aperture pattern and callose deposition in late tetrad stage in three species producing atypical pollen grains.

PREMISE OF THE STUDY: Pollen grains of flowering plants display a fascinating diversity of forms, in spite of their minute size. The observed diversity is determined by the developmental mechanisms implicated in the establishment of pollen morphological features. Pollen grains are generally surrounded by an extremely resistant wall interrupted in places by apertures that play a key role in reproduction, being the places at which pollen tube growth is initiated. Aperture shape, number, and position are determined during microsporogenesis (male meiosis), the earliest step in pollen ontogeny. We investigate in detail the unfolding of microsporogenesis in three species that present uncommon aperture pattern (i.e., disulculate in Calycanthus floridus [Calycanthaceae, magnoliids], tetraporate in Hohenbergia stellata [Bromeliaceae, monocots], and monoporate in Typha latifolia [Typhaceae, monocots]). METHODS: We performed a comparative analysis of microsporogenesis and aperture distribution within tetrads in these species with contrasting aperture arrangements. This was done using aniline blue coloration and UV light microscope observations. KEYS RESULTS: We show that aperture localization and features of callose deposition on intersporal walls produced during cytokinesis coincide in all three species examined. Such a correlation suggests that patterns of callose deposition are strongly involved in determining aperture localization. CONCLUSION: In flowering plants, patterns of male meiosis and especially callose deposition following meiosis may be implicated in the diversity of pollen aperture patterns.

The influence of tetrad shape and intersporal callose wall formation on pollen aperture pattern ontogeny in two eudicot species.

BACKGROUND AND AIMS: In flowering plants, microsporogenesis is accompanied by various types of cytoplasmic partitioning (cytokinesis). Patterns of male cytokinesis are suspected to play a role in the diversity of aperture patterns found in pollen grains of angiosperms. The relationships between intersporal wall formation, tetrad shape and pollen aperture pattern ontogeny are studied. METHODS: A comparative analysis of meiosis and aperture distribution was performed within tetrads in two triporate eudicot species with contrasting aperture arrangements within their tetrads [Epilobium roseum (Onagraceae) and Paranomus reflexus (Proteaceae)]. KEY RESULTS AND CONCLUSIONS: Intersporal wall formation is a two-step process in both species. Cytokinesis is first achieved by the formation of naked centripetal cell plates. These naked cell plates are then covered by additional thick, localized callose deposits that differ in location between the two species. Apertures are finally formed in areas in which additional callose is deposited on the cell plates. The recorded variation in tetrad shape is correlated with variations in aperture pattern, demonstrating the role of cell partitioning in aperture pattern ontogeny.

Formation and function of a new pollen aperture pattern in angiosperms: The proximal sulcus of Tillandsia leiboldiana (Bromeliaceae).

Pollen grains are generally surrounded by an extremely resistant wall interrupted in places by apertures that play a key role in reproduction; pollen tube growth is initiated at these sites. The shift from a proximal to distal aperture location is a striking innovation in seed plant reproduction. Reversals to proximal aperture position have only very rarely been described in angiosperms. The genus Tillandsia belongs to the Bromeliaceae family, and its aperture pattern has been described as distal monosulcate, the most widespread aperture patterns recorded in monocots and basal angiosperms. Here we report developmental and functional elements to demonstrate that the sulcate aperture in Tillandsia leiboldiana is not distal as previously described but proximal. Postmeitotic tetrad observation indicates unambiguously the proximal position of the sulcus, and in vitro germination of pollen grains confirms that the aperture is functional. This is the first report of a sulcate proximal aperture with proximal germination. The observation of microsporogenesis reveals specific features in the patterns of callose thickenings in postmeiotic tetrads.

Microsporogenesis variation in Codiaeum producing inaperturate pollen grain.

A study of microsporogenesis (the earliest stage of pollen ontogeny) was undertaken in seven cultivars of Codiaeum variegatum var. pictum, a eudicot species that produces inaperturate pollen grains. Microsporogenesis appears highly variable for the developmental events suspected to be implicated in the determination of aperture pattern. Most eudicots have tri-aperturate pollen grains and microsporogenesis is described as highly conserved in this clade. The observed burst of variation in C. variegatum therefore appears especially remarkable. A plausible hypothesis to explain the variation is that the pollen being inaperturate, the selective forces applying on the ontogeny of the aperture pattern are relaxed.

Phylogenetic comparative analysis of microsporogenesis in angiosperms with a focus on monocots.

This paper presents the first broad overview of three main features of microsporogenesis (male meiosis) in angiosperms: cytokinesis (cell division), intersporal wall formation, and tetrad form. A phylogenetic comparative approach was used to test for correlated evolution among these characters and to make hypotheses about evolutionary trends in microsporogenesis. The link between features of microsporogenesis and pollen aperture type was examined. We show that the pathway associated with successive cytokinesis (cytoplasm is partitioned after each meiotic division) is restricted to wall formation mediated by centrifugally developing cell plates, and tetragonal (or decussate, T-shaped, linear) tetrads. Conversely, much more flexibility is observed when cytokinesis is simultaneous (two meiotic divisions completed before cytoplasmic partitioning). We suggest that the ancestral type of microsporogenesis for angiosperms, and perhaps for all seed plants, associated simultaneous cytokinesis with centripetal wall formation, resulting in a large diversity in tetrad forms, ranging from regular tetrahedral to tetragonal tetrads, including rhomboidal tetrads. From this ancestral pathway, switches toward successive cytokinesis occurred among basal angiosperms and monocots, generally associated with a switch toward centrifugal intersporal wall formation, whereas eudicots evolved toward an almost exclusive production of regular tetrahedral tetrads. No straightforward link is found between the type of microsporogenesis and pollen aperture type.

Amplification of Nicotiana sylvestris mitochondrial subgenomes is under nuclear control and is associated with phenotypic changes.

We have previously shown the presence in a Nicotiana sylvestris protoplast-derived plant of both a nuclear mutation conferring male sterility (ms4) and a mtDNA reorganisation, named U, characterised by the amplification of substoichiometric mtDNA fragments generated by recombination in the parent T mtDNA. Here we show by physical mapping that the recombining repeats are in direct orientation, thus generating two subgenomes both of which are amplified in the U organisation to the detriment of the parent molecule, and are maintained through sexual reproduction. The nuclear ms4 mutation is likely to have play a role in the shift in mitochondrial molecule equilibrium, as higher levels of recombinant fragments were present in protoplast-derived T calli carrying the ms4 allele than in wild type calli or leaves. The MS4 gene could then lead to conflictual situation. However, subgenomic molecules were counter-selected during the regeneration process, suggesting the existence of different selective pressures in differentiated and non-differentiated cells. The U organisation is associated with higher stem height and late flowering, characters that may not be neutral from a selection point of view. The U equilibrium is an unusual example of sudden mtDNA reorganisation, without obvious differences in genetic information and with only a limited phenotypic impact.

Dynamics of the mitochondrial genome during Podospora anserina aging.

Senescence in Podospora anserina is always correlated with extrachromosomal mitochondrial DNA amplification (senDNA). Here we report a quantitative kinetic analysis of the molecular events that occur in the mitochondrial DNA of several wild-type cultures during aging. For each culture, the amplification of senDNA molecules and the modifications of the mitochondrial chromosome are analyzed at different ages and in relation with two age-related parameters: growth rate and fertility. We find senDNAs exponentially amplified from the germination state and particular regions of the mitochondrial chromosome preferentially lost only in the pre-senescent state, i.e. a few centimeters before the growth arrest of the mycelium. This late loss of information, concomitant with the first phenotypic expression of senescence (loss of fertility), begins in the regions from which the senDNAs originate and culminates with the growth arrest of the mycelium.