Most angiosperm flowers and vegetative shoots of land plants have an acropetal sequence of organ initiation, namely, the more proximal the organ position, the earlier its primordium appears in ontogeny. This is apparently the ancestral condition for angiosperm flowers. An acropetal sequence is the only way in which pre-patterning of organs along a spiral can be established and the only documented sequence of visible organ initiation in spiral flowers. This constraint is absent for whorled flowers, in contrast to whorled vegetative shoots. Non-acropetal developmental sequences are known from both species-rich angiosperm clades with flowers that are predominantly (eudicots) or exclusively whorled (monocots). Deviations from a strict acropetal pattern can be interpreted as sequence heterochronies, related to early termination of the floral meristem and subsequent fractionation of the floral apex into organ primordia. Monocots provide a useful opportunity to generate a large-scale comparative analysis of sequence heterochronies in floral evolution. We recognize several patterns of heterochronic shifts in monocot flowers. An ‘external’ pattern is related to the influence of extrafloral structures such as a flower-subtending bract, creating a delay in initiation of outer whorl organ(s) in the abaxial sector. All ‘internal’ patterns lack any obvious external influence; they include: (1) basipetal organ initiation in wind- and water-pollinated flowers, associated with overall perianth reduction; (2) basipetal organ initiation in secondarily multistaminate androecia (some palms and some Alismataceae) and secondarily multicarpellate gynoecia (some Triuridaceae); (3) non-acropetal patterns associated with common primordia, mostly tepal-stamen (occasional instances in six monocot orders); (4) simultaneous initiation of two neighbouring whorls lacking common primordia (some Alismatales, Dioscoreales, Pandanales, Poales). We propose a typification of common primordia, recognizing tangential and radial types, the latter being either bilobed or entire at the earliest stages. The following hypothesis is testable, but yet untested: at least the entire type is related to the development of two organs from the same prepatterning site. The most important tendencies distinguishing monocots from eudicots are the more frequent occurrence of common primordia, their presence in some epigynous flowers, the scarcity of polymerous androecia with centrifugal development, the apparent absence of complex polyandry, and polyandry associated with a hypanthium. Some monocots have flowers with multicarpellate gynoecia and at the same time oligostaminate androecia, a pattern that is rare in eudicots.

Premise The inclusion of heterotrophic plants in broadly focused systematic studies is challenging, reflecting substantial morphological modification, gene loss, and accelerated nucleotide substitution following photosynthesis loss. In particular, inclusion of very long branch (VLB) taxa with highly elevated rates may lead to phylogenomic misinference. Methods We explore how rapidly evolving plastomes behave in model-based phylogenetic inferences of the yam order Dioscoreales, which experienced convergent photosynthesis losses. The taxon sampling includes all photosynthetic genera, and mycoheterotrophic lineages with a broad range of substitution rates. Results When only moderately rapidly evolving heterotrophs are included, relationships within Dioscoreales are congruent with recent mitochondrial analysis, and generally strongly supported. Photosynthetic Stenomeris (represented by only three genes) is moderately supported as sister to Dioscoreaceae. Notably, Burmanniaceae and Thismiaceae, two mycoheterotrophic families including highly reduced plastomes, are strongly supported as distantly related within Dioscoreales. Constrained analysis rejects combining Thismiaceae with Burmanniaceae, conflicting with current angiosperm classification. Including VLB mycoheterotrophs in phylogenetic inference can lead to localized to broad reduction in branch support—substantially so for Afrothismiaceae, a family placing in disparate locations in Dioscoreales (and beyond) in variant analyses, with no ability to differentiate among its alternative placements. Conclusions Inclusion of highly rate-elevated taxa in analyses of Dioscoreales phylogeny can depress branch support. Inferred relationships in the order are otherwise congruent with studies based on mitochondrial data; the family-level classification needs updating. The extraordinarily rapidly evolving family Afrothismiaceae places inconsistently in diverse plastid analyses, pointing to probable analytical limits for plastid-based phylogenomic analyses of VLB heterotrophs. ### Competing Interest Statement The authors have declared no competing interest. All data matrices are available in figshare ().

PREMISE:New insights into biomineral uptake and sequestration are important for understanding how plants grow. Some plants accumulate silica accretions in precise locations in particular cells. Among monocots, controlled biosilicification occurs in several different forms and is restricted to commelinids and orchids. METHODS:We utilized energy-dispersive x-ray spectroscopy (EDX/EDS) mapping technology on leaf transverse sections to explore the diverse silica deposition patterns in a range of monocots. The results were evaluated using character optimizations on existing phylogenies. RESULTS:Our optimization indicates at least two independent evolutionary origins of phytoliths among monocots, with secondary losses in some lineages. Silica that accumulates in the cell lumen occurs mostly in bundle sheath cells or epidermal cells, often associated with sclerenchyma. In Bromeliaceae and Rapateaceae, small phytoliths occur in the walls of occluded epidermal cells overlying sclerenchyma. In Dasypogonaceae, phytoliths accumulate in the lumen of epidermal cells. Cell-wall bound silica occurs in the epidermal cells of some commelinids (Commelinaceae, Cyperaceae and Poaceae). There is a close association between silica deposition and the presence of ferulic acid, except possibly in orchids. Records of high silica concentration in leaves are not always correlated with deposition. We found no silica deposition in leaves of some aquatic commelinids, despite evidence for silica uptake and presence of ferulic acid. CONCLUSIONS:Our ongoing comparative investigations using EDX data not only extend our knowledge about biomineral inclusions in plants, but also highlight their structural and biochemical complexity. This study suggests that the diversity and relatively restricted phylogenetic distribution of monocot phytoliths is at least partly attributable to cell chemistry.

SummaryAfrothismia is a genus of non-photosynthetic mycoheterotrophs from the forests of continental tropical Africa. Multiple phylogenetic inferences using molecular data recover the genus as sister to a clade comprising mycoheterotrophic Thismiaceae and the photosynthetic family Taccaceae, contrary to earlier placements of Afrothismia and Thismiaceae within Burmanniaceae. Morphological support for separating Afrothismia from the rest of Thismiaceae has depended on the zygomorphic flowers of Afrothismia (although some species of Thismia are also zygomorphic), and their clusters of root tubers, each with a terminal rootlet. The number of described species of Afrothismia has recently increased from four to 16, with seven more species as yet undescribed; these discoveries have added morphological characters that support its distinction from Thismiaceae. Most notably, the ovary in Afrothismia has a single stalked placenta, and circumscissile fruits from which seeds are exserted by placental elevation (vs in Thismiaceae, three placentas, a deliquescing fruit lid, and seeds not exserted). Afrothismia stamens are inserted in the lower part of the floral tube, where they are attached to the stigma, and individual flowers are subtended by a single large dorsal bract. In contrast, in Thismiaceae, stamens are inserted at the mouth of the tube, free of and distant from the stigma, and each flower is subtended by a loose whorl of (2 –) 3 (– 4) bracts. Here we formally characterise Afrothismiaceae and review what is known of its development, seed germination, interactions with mycorrhizal Glomeromycota, biogeography, phylogeny and pollination biology. All but one (Afrothismia insignis; Vulnerable) of the 13 species assessed on the IUCN Red List of Threatened Species are either Endangered or Critically Endangered; one species (A. pachyantha) is considered extinct.

Flowers are deceptively simple structures, characterized by a determinate primary axis that bears organs in condensed concentric zones in a strict structural and temporal sequence. Few species have escaped these constraints, but those that have can provide insights into the evolutionary history of flowers if placed in the appropriate phylogenetic and developmental context. For my flowering image, I selected a longitudinal section of a Lacandonia flower, which breaks a fundamental rule of spatial arrangement: the flowers are ‘inside-out’, with the carpels surrounding the stamens—a pattern that is almost unique among angiosperms. When viewed in the context of the family and order to which it belongs, this species has led me into many fascinating areas of comparative and evolutionary plant morphology.

Background and Aims Structural colour is responsible for the remarkable metallic blue colour seen in the leaves of several plants. Species belonging to only ten genera have been investigated to date, revealing four photonic structures responsible for structurally coloured leaves. One of these is the helicoidal cell wall, known to create structural colour in the leaf cells of five taxa. Here we investigate a broad selection of land plants to understand the phylogenetic distribution of this photonic structure in leaves.Methods We identified helicoidal structures in the leaf epidermal cells of 19 species using transmission electron microscopy. Pitch measurements of the helicoids were compared with the reflectance spectra of circularly polarized light from the cells to confirm the structure-colour relationship.Results By incorporating species examined with a polarizing filter, our results increase the number of taxa with photonic helicoidal cell walls to species belonging to at least 35 genera. These include 19 monocot genera, from the orders Asparagales (Orchidaceae) and Poales (Cyperaceae, Eriocaulaceae, Rapateaceae) and 16 fern genera, from the orders Marattiales (Marattiaceae), Schizaeales (Anemiaceae) and Polypodiales (Blechnaceae, Dryopteridaceae, Lomariopsidaceae, Polypodiaceae, Pteridaceae, Tectariaceae).Conclusions Our investigation adds considerably to the recorded diversity of plants with structurally coloured leaves. The iterative evolution of photonic helicoidal walls has resulted in a broad phylogenetic distribution, centred on ferns and monocots. We speculate that the primary function of the helicoidal wall is to provide strength and support, so structural colour could have evolved as a potentially beneficial chance function of this structure.

Despite (or perhaps because of) intensive multidisciplinary research, opinions on the optimal number of species recognised within the Eurasian orchid genus Ophrys range from nine to at least 400. The lower figure of nine macrospecies is based primarily on seeking small but reliable discontinuities in DNA ‘barcode’ regions, an approach subsequently reinforced and finessed via high-throughput sequencing studies. The upper figure of ca. 400 microspecies reflects the morphological authoritarianism of traditional taxonomy combined with belief in extreme pollinator specificity caused by reliance on pollination through pseudo-copulation, enacted by bees and wasps. Groupings of microspecies that are less inclusive than macrospecies are termed mesospecies. Herein, we present multivariate morphometric analyses based on 51 characters scored for 457 individual plants that together span the full morphological and molecular diversity within the genus Ophrys, encompassing 113 named microspecies that collectively represent all 29 mesospecies and all nine macrospecies. We critique our preferred morphometric approach of accumulating heterogeneous data and analysing them primarily using principal coordinates, noting that our conclusions would have been strengthened by even greater sampling and the inclusion of data describing pseudo-pheromone cocktails. Morphological variation within Ophrys proved to be exceptionally multidimensional, lacking strong directional trends. Multivariate clustering of plants according to prior taxonomy was typically weak, irrespective of whether it was assessed at the level of macrospecies, mesospecies or microspecies; considerable morphological overlap was evident even between subsets of the molecularly differentiable macrospecies. Characters supporting genuine taxonomic distinctions were often sufficiently subtle that they were masked by greater and more positively correlated variation that reflected strong contrasts in flower size, tepal colour or, less often, plant size. Individual macrospecies appear to represent morphological continua, within which taxonomic divisions are likely to prove arbitrary if based exclusively on morphological criteria and adequately sampled across their geographic range. It remains unclear how much of the mosaic of subtle character variation among the microspecies reflects genetic versus epigenetic or non-genetic influences and what proportion of any contrasts observed in gene frequencies can be attributed to the adaptive microevolution that is widely considered to dictate speciation in the genus. Moreover, supplementing weak morphological criteria with extrinsic criteria, typically by imposing constraints on geographic location and/or supposed pollinator preference, assumes rather than demonstrates the presence of even the weakest of species boundaries. Overall, it is clear that entities in Ophrys below the level of macrospecies have insufficiently structured variation, either phenotypic or genotypic, to be resolved into discrete, self-circumscribing (“natural”) entities that can legitimately be equated with species as delimited within other less specialised plant genera. Our search for a non-arbitrary (meso)species concept competent to circumscribe an intermediate number of species has so far proven unsuccessful.

Summary Structural color is poorly known in plants relative to animals. In fruits, only a handful of cases have been described, including in Viburnum tinus where the blue color results from a disordered multilayered reflector made of lipid droplets. Here, we examine the broader evolutionary context of fruit structural color across the genus Viburnum. We obtained fresh and herbarium fruit material from 30 Viburnum species spanning the phylogeny and used transmission electron microscopy, optical simulations, and ancestral state reconstruction to identify the presence/absence of photonic structures in each species, understand the mechanism producing structural color in newly identified species, relate the development of cell wall structure to reflectance in Viburnum dentatum, and describe the evolution of cell wall architecture across Viburnum. We identify at least two (possibly three) origins of blue fruit color in Viburnum in species which produce large photonic structures made of lipid droplets embedded in the cell wall and which reflect blue light. Examining the full spectrum of mechanisms producing color in pl, including structural color as well as pigments, will yield further insights into the diversity, ecology, and evolution of fruit color.

The magnoliid clade encompasses 18 extant families arranged in four orders, plus several extinct taxa, including some of the most ancient angiosperm fossils. The clade is characterized by paracytic stomata with a distinct pair of lateral subsidiary cells that flank the guard cells, though other stomatal types are also reported, including anomocytic and anisocytic. In contrast with monocots, the paracytic stomata of magnoliids develop from linear triads, and the lateral subsidiary cells are stomatal-lineage ground cells (SLGCs). Anisocytic stomata typically possess three SLGCs. Amplifying divisions are rare in magnoliids, but occur in some Piperales, in association with anisocytic stomata. Differences in mature stomatal types result from differences in cell shape and polarity at critical developmental stages. Stomatal clusters have been reported in Cinnamomum (Lauraceae) and Galbulimima (Himantandraceae), but neither are apparently formed by amplifying divisions, in contrast with eudicots. In Galbulimima , each peltate scale hair is surrounded by a ring of 3–8 non-contiguous stomata, each derived from different initial meristemoids.

IntroductionUnderstanding the complex inflorescence architecture and developmental morphology of common buckwheat (Fagopyrum esculentum) is crucial for crop yield. However, most published descriptions of early flower and inflorescence development in Polygonaceae are based on light microscopy and often documented by line drawings. In Fagopyrum and many other Polygonaceae, an important inflorescence module is the thyrse, in which the primary axis never terminates in a flower and lateral cymes (monochasia) produce successively developing flowers of several orders. Each flower of a cyme is enclosed together with the next-order flower by a bilobed sheathing bract-like structure of controversial morphological nature.MethodsWe explored patterns of flower structure and arrangement in buckwheat and its wild relatives, using comparative morphology, scanning electron microscopy and X-ray microtomography.ResultsOur data support interpretation of the sheathing bract as two congenitally fused phyllomes (prophylls), one of which subtends a next-order flower. In tepal-like bract, a homeotic mutant of F. esculentum, the bilobed sheathing bract-like organ acquires tepal-like features and is sometimes replaced by two distinct phyllomes. Wild representatives of F. esculentum (ssp. ancestrale) and most cultivars of common buckwheat possess an indeterminate growth type with lateral thyrses produced successively on the primary inflorescence axis until cessation of growth. In contrast, determinate cultivars of F. esculentum develop a terminal thyrse after producing lateral thyrses. In contrast to F. esculentum, the occurrence of a terminal thyrse does not guarantee a determinate growth pattern in F. tataricum. The number of lateral thyrses produced before the terminal thyrse on the main axis of F. tataricum varies from zero to c. 19.DiscussionThe nine stages of early flower development formally recognized here and our outline of basic terminology will facilitate more standardized and readily comparable descriptions in subsequent research on buckwheat biology. Non-trivial relative arrangements of tepals and bracteoles in Fagopyrum and some other Polygonaceae require investigation using refined approaches to mathematical modelling of flower development. Our data on inflorescence morphology and development suggest contrasting evolutionary patterns in the two main cultivated species of buckwheat, F. esculentum and F. tataricum. The genus Fagopyrum offers an excellent opportunity for evo-devo studies related to inflorescence architecture.

BACKGROUND:Oriented patterning of epidermal cells is achieved primarily by transverse protodermal cell divisions perpendicular to the organ axis, followed by axial cell elongation. In linear leaves with parallel venation, most stomata are regularly aligned with the veins. This longitudinal patterning operates under a strong developmental constraint and has demonstrable physiological benefits, especially in grasses. However, transversely oriented stomata characterize a few groups, among both living angiosperms and extinct Mesozoic seed plants.SCOPE:This review examines comparative and developmental data on stomatal patterning in a broad phylogenetic context, focusing on the evolutionary and ecophysiological significance of guard-cell orientation. It draws from a diverse range of literature to explore the pivotal roles of the plant growth hormone auxin in establishing polarity and chemical gradients that enable cellular differentiation.CONCLUSIONS:Transverse stomata evolved iteratively in a few seed-plant groups during the Mesozoic era, especially among parasitic or xerophytic taxa, such as the hemiparasitic mistletoe genus Viscum and the xerophytic shrub Casuarina, indicating a possible link with ecological factors such as the Cretaceous CO2 decline and changing water availability. The discovery of this feature in some extinct seed-plant taxa known only from fossils could represent a useful phylogenetic marker.

PREMISE:The sporoderm of seed-plant pollen grains typically has apertures in which the outer sporopollenin-bearing layer is relatively sparse. The apertures allow regulation of the internal volume of the pollen grain during desiccation and rehydration (harmomegathy) and also serve as sites of pollen germination. A small fraction of angiosperms undergo pollination in water or at the water surface, where desiccation is unlikely. Their pollen grains commonly lack apertures, though with some notable exceptions. We tested a hypothesis that in some angiosperm aquatics that inhabit water of unstable salinity, the pollen apertures accommodate osmotic effects that occur during pollination in such conditions.METHODS:Pollen grains of the tepaloid clade of the monocot order Alismatales, which contains ecologically diverse aquatic and marshy plants, were examined using light microscopy and scanning electron microscopy. We used Ruppia as a model to test pollen grain response in water of various salinities. Pollen aperture evolution was also analyzed using molecular tree topologies.RESULTS:Phylogenetic optimizations demonstrated an evolutionary loss and two subsequent regains of the aperturate condition in the tepaloid clade of Alismatales. Both of the taxa that have reverted to aperturate pollen (Ruppia, Ruppiaceae; Althenia, Potamogetonaceae) are adapted to changeable water salinity. Direct experiments with Ruppia showed that the pollen apertures have a role in a harmomegathic response to differences in water salinity.CONCLUSIONS:Our results showed that the inferred regain of pollen apertures represents an adaptation to changeable water salinity. We invoke a loss-and-regain scenario, prompting questions that are testable using developmental genetics and plant physiology.

The family Rapateaceae represents an early-divergent lineage of Poales with biotically pollinated showy flowers. We investigate developmental morphology and anatomy in all three subfamilies and five tribes of Rapateaceae to distinguish between contrasting hypotheses on spikelet morphology and to address questions on the presence of nectaries and gynoecium structure. We support an interpretation of the partial inflorescence (commonly termed spikelet), as a uniaxial system composed of a terminal flower and numerous empty phyllomes. A terminal flower in an inflorescence unit is an autapomorphic feature of Rapateaceae. The gynoecium consists of synascidiate, symplicate, and usually asymplicate zones, with gynoecium formation encompassing congenital and often also postgenital fusions between carpels. Species of Rapateaceae differ in the relative lengths of the gynoecial zones, the presence or absence of postgenital fusion between the carpels and placentation in the ascidiate or plicate carpel zones. In contrast with previous reports, septal nectaries are lacking in all species. The bird-pollinated tribe Schoenocephalieae is characterized by congenital syncarpy; it displays an unusual type of gynoecial (non-septal) nectary represented by a secretory epidermis at the gynoecium base.

Some of the earliest diverging flowers have pollinator-attracting conical cells, indicating that the genetic pathway controlling their development is also ancient. Conical epidermal cells occur on the tepals (perianth organs, typically petals and/or sepals) of the majority of animal-pollinated angiosperms, where they play both visual and tactile roles in pollinator attraction, providing grip to foraging insects, and enhancing colour, temperature, and hydrophobicity. To explore the evolutionary history of conical epidermal cells in angiosperms, we surveyed the tepal epidermis in representative species of the ANA-grade families, the early-diverging successive sister lineages to all other extant angiosperms, and analysed the function of a candidate regulator of cell outgrowth from Cabomba caroliniana (Nymphaeales). We identified conical cells in at least two genera from different families (Austrobaileya and Cabomba). A single SBG9 MYB gene was isolated from C. caroliniana and found to induce strong differentiation of cellular outgrowth, including conical cells, when ectopically expressed in Nicotiana tabacum. Ontogenetic analysis and quantitative reverse transcription-PCR established that CcSBG9A1 is spatially and temporally expressed in a profile which correlates with a role in conical cell development. We conclude that conical or subconical cells on perianth organs are ancient within the angiosperms and most probably develop using a common genetic programme initiated by a SBG9 MYB transcription factor.

EDITORIAL article Front. Plant Sci., 10 November 2022Sec. Plant Development and EvoDevo https://doi.org/10.3389/fpls.2022.1076169

Plants combine both chemical and structural means to appear colorful. We now have an extensive understanding of the metabolic pathways used by flowering plants to synthesize pigments, but the mechanisms remain obscure whereby cells produce microscopic structures sufficiently regular to interfere with light and create an optical effect. Here, we combine transgenic approaches in a novel model system, Hibiscus trionum, with chemical analyses of the cuticle, both in transgenic lines and in different species of Hibiscus, to investigate the formation of a semi-ordered diffraction grating on the petal surface. We show that regulating both cuticle production and epidermal cell growth is insufficient to determine the type of cuticular pattern produced. Instead, the chemical composition of the cuticle plays a crucial role in restricting the formation of diffraction gratings to the pigmented region of the petal. This suggests that buckling, driven by spatiotemporal regulation of cuticle chemistry, could pattern the petal surface at the nanoscale.

Molecular phylogenetic analyses have revealed a superclade of mesangiosperms with five extant lineages: monocots, eudicots, magnoliids, Ceratophyllum and Chloranthaceae. Both Ceratophyllum and Chloranthaceae are ancient lineages with a long fossil record; their precise placement within mesangiosperms is uncertain. Morphological studies have suggested that they form a clade together with some Cretaceous fossils, including Canrightia , Montsechia and Pseudoasterophyllites . Apart from Canrightia , members of this clade share unilocular gynoecia commonly interpreted as monomerous with ascidiate carpels. Alternatively, the gynoecium of Ceratophyllum has also been interpreted as syncarpous with a single fertile carpel (pseudomonomerous). We investigate patterns of morphological, anatomical and developmental variation in gynoecia of three Ceratophyllum species to explore the controversial interpretation of its gynoecium as either monomerous or pseudomonomerous. We use an angiosperm-wide morphological data set and contrasting tree topologies to estimate the ancestral gynoecium type in both Ceratophyllum and mesangiosperms. Gynoecia of all three Ceratophyllum species possess a small (sometimes vestigial) glandular appendage on the abaxial side and an occasionally bifurcating apex. The ovary is usually unilocular with two procambium strands, but sometimes bilocular and/or with three strands in C. demersum . None of the possible phylogenetic placements strongly suggest apocarpy in the stem lineage of Ceratophyllum . Rescoring Ceratophyllum as having two united carpels affects broader-scale reconstructions of the ancestral gynoecium in mesangiosperms. Our interpretation of the glandular appendage as a tepal or staminode homologue makes the Ceratophyllum ovary inferior, thus resembling (semi)inferior ovaries of most Chloranthaceae and potentially related fossils Canrightia and Zlatkocarpus . The entire structure of the flower of Ceratophyllum suggests strong reduction following a long and complex evolutionary history. The widely accepted notion that apocarpy is ancestral in mesangiosperms (and angiosperms) lacks robust support, regardless of which modes of carpel fusion are considered. Our study highlights the crucial importance of incorporating fossils into large-scale analyses to understand character evolution.

SummaryKew’s Jodrell Laboratory was established in 1876 as a centre for botanical research in disciplines including plant physiology, anatomy and embryology, palaeobotany and mycology. Despite relatively little available funding, its location in one of the world’s largest botanic gardens and close to several well-curated plant collections has ensured its continued existence for almost a century and a half. Under the far-sighted leadership of Kew’s second Director, Joseph Dalton Hooker, the Jodrell Laboratory was established to coincide with Thomas Henry Huxley’s pioneering course at the Normal School of Science in London. Funded by a generous private donation, the Laboratory complemented and augmented the programme in taxonomy and systematics already established in Kew’s Herbarium, and provided a broader educational and research base to explore contemporary laboratory-based discoveries in fields such as physiology and lifecycles (sometimes termed the “New Botany”). The Jodrell Laboratory represents one of the world’s first non-university affiliated laboratories and has spawned several “spin-off” facilities such as the Laboratory of Plant Pathology and the Millennium Seed Bank. This paper traces its early influence as an important centre for research in palaeobotany and plant systematics, its subsequent decline during the inter-war years, and a relatively dynamic period of innovative research following the construction of a new building on the same site.