Reconstruction of an enigmatic Pennsylvanian cone reveals a relationship to Sphenophyllales.

PREMISE: We studied the 3D morphology of a small, well-preserved cone from the Pennsylvanian Mazon Creek Lagerstätte to characterize its structure and determine its systematic affinity. Previously tentatively assigned to the enigmatic Tetraphyllostrobus, we show that it differs in key respects from that genus as described. METHODS: We systematically compared the new fossil with relevant Paleozoic cone genera and employed advanced imaging techniques, including scanning electron microscopy, Airyscan confocal super-resolution microscopy, optical microscopy, and X-ray microcomputed tomography to visualize and reconstruct the fossil cone in 3D. RESULTS: The analyses demonstrate unequivocally that the sporophylls of the new Mazon Creek cone are arranged in whorls of six and have characters typical of Sphenophyllales, including epidermal cells with undulatory margins and in situ spores assignable to Columinisporites. The combination of characters, including sporophyll arrangement, anatomy, and spore type, supports the establishment of Hexaphyllostrobus kostorhysii gen. et sp. nov. within Sphenophyllales. Furthermore, we show that Tetraphyllostrobus, although originally described as possessing smooth monolete spores, actually possesses Columinisporites-type spores, indicating that it, too, was most likely a sphenophyll. CONCLUSIONS: The recognition of Hexaphyllostrobus contributes to our knowledge of Pennsylvanian sphenophyll diversity, and in particular increases the number of species with in situ Columinisporites-type spores. Attribution of Hexaphyllostrobus to Sphenophyllales calls into question current interpretations of Tetraphyllostrobus suggesting that future research on better-preserved macrofossil material may demonstrate a sphenophyllalean relationship.

Plant periderm as a continuum in structural organisation: a tracheophyte-wide survey and hypotheses on evolution.

Periderm is a well-known structural feature with vital roles in protection of inner plant tissues and wound healing. Despite its importance to plant survival, knowledge of periderm occurrences outside the seed plants is limited and the evolutionary origins of periderm remain poorly explored. Here, we review the current knowledge of the taxonomic distribution of periderm in its two main forms - canonical periderm (periderm formed as a typical ontogenetic stage) and wound periderm (periderm produced as a self-repair mechanism) - with a focus on major plant lineages, living and extinct. We supplement the published occurrences with data based on our own observations and experiments. This updated body of data reveals that the distribution of wound periderm is more widespread taxonomically than previously recognized and some living and extinct groups are capable of producing wound periderm, despite canonical periderm being absent from their normal developmental program. A critical review of canonical and wound periderms in extant and fossil lineages indicates that not all periderms are created equal. Their organisation is widely variable and the differences can be characterised in terms of variations in three structural features: (i) the consistency in orientation of periclinal walls within individual files of periderm cells; (ii) the lateral coordination of periclinal walls between adjacent cell files; and (iii) whether a cambial layer and conspicuous layering of inward and outward derivatives can be distinguished. Using a new system of scoring periderm structure based on these criteria, we characterise the level of organisation of canonical and wound periderms in different lineages. Looking at periderms through the lens provided by their level of organisation reveals that the traditional image of periderm as a single generalised feature, is best viewed as a continuum of structural configurations that are all predicated by the same basic process (periclinal divisions), but can fall anywhere between very loosely organized (diffuse periclinal growth) to very tightly coordinated (organized periclinal growth). Overall, wound periderms in both seed plants and seed-free plants have lower degrees of organisation than canonical periderms, which may be due to their initiation in response to inherently disruptive traumatic events. Wound and canonical periderms of seed plants have higher degrees of organisation than those of seed-free plants, possibly due to co-option of the programs responsible for organizing their vascular cambial growth. Given the importance of wound periderm to plant survival, its widespread taxonomic distribution, and its early occurrence in the fossil record, we hypothesise that wound periderm may have had a single origin in euphyllophytes and canonical periderm may have originated separately in different lineages by co-option of the basic regulatory toolkit of wound periderm formation. In one evolutionary scenario, wound periderm regulators activated initially by tissue tearing due to tensional stresses elicited by woody growth underwent heterochronic change that switched their activation trigger from tissue tearing to the tensional stresses that precede it, with corresponding changes in the signalling that triggered the regulatory cascade of periderm development from tearing-induced signals to signalling induced by tension in cells.

Detailed seed cone morpho-anatomy of the Prumnopityoid clade: an insight into the origin and evolution of Podocarpaceae seed cones.

BACKGROUND AND AIMS: Seed cone traits are significant for understanding the evolutionary history of conifers. Podocarpaceae has fleshy cones with a distinct morphology compared with other conifers. However, we have a poor understanding of the seed cone morphology of the Prumnopityoid clade and within Podocarpaceae. This study presents detailed seed cone morpho-anatomy and the evolution of fleshy structures traits in the Prumnopityoid clade. METHODS: We investigated the detailed seed cone morpho-anatomy of selected species from the nine genera using the histological method. The evolution of morpho-anatomical traits was assessed using ancestral state reconstruction methods. KEY RESULTS: The Prumnopityoid clade has evolved fleshy seed cones using different functional structures (e.g. aril, epimatium, bracts or receptaculum) and fleshiness is an ancestral trait in the clade. An epimatium is present in all genera except Phyllocladus, but with different structural morphologies (e.g. a fleshy asymmetrical cup-like epimatium or an epimatium that is fused with the integument, forming a fleshy sarcotesta-like seed coat). In all species with fleshy sarcotesta-like seed coats, the endotesta is hard and woody, forming a sclerotesta-like structure and the epimatium and exotesta are fused, forming a fleshy sarcotesta-like structure. CONCLUSIONS: This study highlights that the Prumnopityoid clade has an amazing diversity of structures and complex evolutionary patterns. Fleshiness is an ancestral trait of the clade and has been achieved via diverse evolutionary pathways and structures. This clade has four distinct seed cone types, i.e. drupe-like, receptaculate, arilloid and dacrydioid cones, based on morpho-anatomical structures and traits. The macrofossil record also demonstrates the presence of several structures and traits.

Evolution of the coniferous seed scale.

BACKGROUND: The Florin model is the commonly accepted theory of coniferous seed scale evolution. It describes the derivation of extant seed scale morphology from the morphology of fossil conifers via the reduction of complex to simple axillary structures. In this framework the seed scale is composed of a reduced lateral shoot with fertile and sterile appendages which are interpreted as leaf homologues. SCOPE: The Florin model has three crucial problems that we address here: (1) the original derivation series does not take the ontogeny of extant conifers into account, (2) it cannot explain the morphology of all extant conifers and (3) Taxaceae were originally excluded. Examination of seed cones of extant conifers shows that ovules occur in three different positions in the cone: in an axillary position, replacing a leaf or terminating the cone axis. By interpreting the fertile appendage or seed-bearing structure as a leaf, not all positions are possible. The exclusion of Taxaceae from conifers is in stark contrast to recent molecular phylogenetic studies, which include Taxaceae in conifers as sister to Cupressaceae. Therefore, the Florin model does not offer an adequate explanation for taxaceous morphology. CONCLUSION: We conclude that the seed-bearing structure of conifers cannot be interpreted as homologous to a leaf. In the interpretation we present here, the seed-bearing structure is the modified funiculus of the ovule, multiples of which laterally fuse to form the seed scale. The seed scales of all extant conifers can be derived from a Cunninghamia-like morphology via fusion and reduction of individual funiculi.

Cavitation fatigue in conifers: a study on eight European species.

After drought-induced embolism and repair, tree xylem may be weakened against future drought events (cavitation fatigue). As there are few data on cavitation fatigue in conifers available, we quantified vulnerability curves (VCs) after embolism/repair cycles on eight European conifer species. We induced 50% and 100% loss of conductivity (LC) with a cavitron, and analyzed VCs. Embolism repair was obtained by vacuum infiltration. All species demonstrated complete embolism repair and a lack of any cavitation fatigue after 50% LC . After 100% LC, European larch (Larix decidua), stone pine (Pinus cembra), Norway spruce (Picea abies), and silver fir (Abies alba) remained unaffected, while mountain pine (Pinus mugo), yew (Taxus baccata), and common juniper (Juniperus communis) exhibited 0.4-0.9 MPa higher vulnerability to embolism. A small cavitation fatigue observed in Scots pine (Pinus sylvestris) was probably biased by incomplete embolism repair, as indicated by a correlation of vulnerability shifts and conductivity restoration. Our data demonstrate that cavitation fatigue in conifers is species-specific and depends on the intensity of preceding LC. The lack of fatigue effects after moderate LC, and relevant effects in only three species after high LC, indicate that conifers are relatively resistant against cavitation fatigue. This is remarkable considering the complex and delicate conifer pit architecture and may be important considering climate change projections.

Vascular plants on inselberg landscapes in Espírito Santo state: bases for the creation of a protected area in southeastern Brazil

Inselbergs are granitic or gneissic rocky outcrops prevalent in the landscape of southeastern Brazil. These ecosystems represent islands of isolated habitats that harbor a peculiar flora with high richness and endemism. The present study lists the species of vascular plants occurring in the Pedra da Andorinha Complex, located in the municipality of Jerônimo Monteiro/Cachoeiro de Itapemirim, southern Espírito Santo state, aiming to generate subsidies for the creation of a protected area. The survey was performed between July 2017 and October 2018, resulting in a record of 121 species, 96 genera, and 40 families. Bromeliaceae (17), Orchidaceae (12) and Fabaceae (10) were the richest families. The phytophysiognomy of exposed rock vegetation comprises a greater number of species (79 species) compared to the woody rupicolous communities (42). Eighteen of the collected species are threatened by extinction; a new species was discovered; and five were described based on materials previously collected in the studied location — Alcantarea patriae, Anthurium martinellii, Coleocephalocereus uebelmanniorum, Stigmatodon attenuatoides and Pitcairnia azouryi, the first four being endemic to the region. We also found Tabebuia reticulata, a rare species among Brazilian flora. Our results highlight the biological importance of the Pedra das Andorinhas Complex and reinforce the need to create a protected area to preserve biodiversity and the regional natural heritage.

Gymnosperm glandular trichomes: expanded dimensions of the conifer terpenoid defense system.

Glandular trichomes (GTs) are defensive structures that produce and accumulate specialized metabolites and protect plants against herbivores, pathogens, and abiotic stress. GTs have been extensively studied in angiosperms for their roles in defense and biosynthesis of high-value metabolites. In contrast, trichomes of gymnosperms have been described in fossilized samples, but have not been studied in living plants. Here, we describe the characterization of GTs on young stems of a hybrid white spruce. Metabolite and histological analysis of spruce GTs support a glandular function with accumulation of a diverse array of mono-, sesqui- and diterpenes including diterpene methylesters. Methylated diterpenes have previously been associated with insect resistance in white spruce. Headspeace analysis of spruce GTs showed a profile of volatiles dominated by monoterpenes and a highly diverse array of sesquiterpenes. Spruce GTs appear early during shoot growth, prior to the development of a lignified bark and prior to accumulation of terpenes in needles. Spruce GTs may provide an early, terpene-based chemical defense system at a developmental stage when young shoots are particularly vulnerable to foliage and shoot feeding insects, and before the resin duct system characteristic of conifers has fully developed.

Community Assembly and Climate Mismatch in Late Quaternary Eastern North American Pollen Assemblages.

Plant community response to climate change ranges from synchronous tracking to strong mismatch. Explaining this variation in climate change response is critical for accurate global change modeling. Here we quantify how closely assemblages track changes in climate (match/mismatch) and how broadly climate niches are spread within assemblages (narrow/broad ecological tolerance, or "filtering") using data for the past 21,000 years for 531 eastern North American fossil pollen assemblages. Although climate matching has been strong over the last 21 millennia, mismatch increased in 30% of assemblages during the rapid climate shifts between 14.5 and 10 ka. Assemblage matching rebounded toward the present day in 10%-20% of assemblages. Climate-assemblage mismatch was greater in tree-dominated and high-latitude assemblages, consistent with persisting populations, slower dispersal rates, and glacial retreat. In contrast, climate matching was greater for assemblages comprising taxa with higher median seed mass. More than half of the assemblages were climatically filtered at any given time, with peak filtering occurring at 8.5 ka for nearly 80% of assemblages. Thus, vegetation assemblages have highly variable rates of climate mismatch and filtering over millennial scales. These climate responses can be partially predicted by species' traits and life histories. These findings help constrain predictions for plant community response to contemporary climate change.

Reconstructing Krassilovia mongolica supports recognition of a new and unusual group of Mesozoic conifers.

Previously unrecognized anatomical features of the cone scales of the enigmatic Early Cretaceous conifer Krassilovia mongolica include the presence of transversely oriented paracytic stomata, which is unusual for all other extinct and extant conifers. Identical stomata are present on co-occurring broad, linear, multiveined leaves assigned to Podozamites harrisii, providing evidence that K. mongolica and P. harrisii are the seed cones and leaves of the same extinct plant. Phylogenetic analyses of the relationships of the reconstructed Krassilovia plant place it in an informal clade that we name the Krassilovia Clade, which also includes Swedenborgia cryptomerioides-Podozamites schenkii, and Cycadocarpidium erdmanni-Podozamites schenkii. All three of these plants have linear leaves that are relatively broad compared to most living conifers, and that are also multiveined with transversely oriented paracytic stomata. We propose that these may be general features of the Krassilovia Clade. Paracytic stomata, and other features of this new group, recall features of extant and fossil Gnetales, raising questions about the phylogenetic homogeneity of the conifer clade similar to those raised by phylogenetic analyses of molecular data.

Assessing the effects of water quality on leaf morphoanatomy, ultrastructure and photosynthetic pigment content of Salvinia auriculata Aubl. (Salviniaceae).

The objective of this study is to evaluate the effects of physical-chemical and biological variables of the water of the Capibaribe River (state of Pernambuco, Brazil) on leaf anatomy, including ultrastructure and photosynthetic pigment of Salvinia auriculata. Specimens of S. auriculata collected in the Gurjaú River, an area with a low pollution degree, were acclimatized in Hoagland's solution and then subjected to three water samples of the Capibaribe River with different levels of pollution. Twenty-one physical-chemical and biological variables were analyzed according to the Standard Methods for the Examination of Water and Wastewater. The results showed that the samples of the Capibaribe River presented nine parameters that did not comply with the current Brazilian legislation. After 15 days of bioassay, S. auriculata presented variations in mesophyll and cuticle thickness, changes in trichome morphology and accumulation of phenolic compounds. No significant differences were observed for photosynthetic pigment content and leaf length of S. auriculata. Multivariate analyses (PCA and Cluster) showed that the point in the Capibaribe River with the highest number of variables that do not comply with the current legislation was responsible for major structural and chemical changes observed in S. auriculata.

Growth, wood anatomy and stable isotopes show species-specific couplings in three Mexican conifers inhabiting drought-prone areas.

An improved understanding of how tree species will respond to warmer conditions and longer droughts requires comparing their responses across different environmental settings and considering a multi-proxy approach. We used several traits (tree-ring width, formation of intra-annual density fluctuations - IADFs, wood anatomy, Δ13C and δ18O records) to retrospectively quantify these responses in three conifers inhabiting drought-prone areas in northwestern Mexico. A fir species (Abies durangensis) was studied in a higher altitude and slightly rainier site and two pine species were sampled in a nearby, lower drier site (Pinus engelmannii, Pinus cembroides). Tree-ring-width indices (TRWi) of the studied species showed a very similar year-to-year variability likely indicating a common climatic signal. Wood anatomy analyses done over 3.5 million measured cells, showed that P. cembroides lumen area was much smaller than in the other two species and it remained constant along all the studied period (over 64 years). Instead, cell wall thickness was widest in P. engelmannii and this species presented the highest amount of intra-annual density fluctuations. Climate and wood anatomy correlations pointed out that lumen area was positively affected by winter precipitation for all studied species, while cell-wall thickness was negatively affected by this season's precipitation in all species but P. cembroides. Stable isotope analysis showed significantly lower values of Δ13C for P. cembroides and no significant δ18O differences between the three species, although they shared a common decreasing trend. With very distinct wood anatomical traits (smaller cells, compact morphology), P. cembroides stood out as the better adapted species in its current environment and could be less affected by future drier climate. P. engelmannii and A. durangensis showed high plasticity at wood anatomical level, allowing them to promptly respond to seasonal water availability but likely gives few advantages on future climate scenarios with longer and frequent drought spells.

The Most Extensive Devonian Fossil Forest with Small Lycopsid Trees Bearing the Earliest Stigmarian Roots.

Since the Late Paleozoic, forests have become distributed worldwide and significantly changed the Earth's climate and landscapes, but the record of forests is rare in the Devonian (419-359 Ma in age) when they first appeared. From the Upper Devonian (Famennian with the age of 372-359 Ma) of Xinhang, Anhui, China, we report a very large in situ forest, which includes locally dense stands of lycopsid plants. The Xinhang forest is monospecific with a small tree lycopsid Guangdedendron gen. nov., probably dioecious with monocarpic reproduction. The plant shows the earliest stigmarian rooting system typical of giant tree lycopsids dominating Carboniferous forests. It colonizes coastal clastic wetlands that were influenced by floods. This significantly increases the paleogeographical coverage of in situ Devonian forests, and contributes to our understanding of atmospheric CO2 decline and coastal consolidation.

Taxonomic study of Triblemma by using light and scanning electron microscopy.

Triblemma Ching is a genus proposed by Ching Ren-Chang in 1978, and it is composed of two species, Triblemma lancea (Thunb.) Ching and Triblemma zeylanica (Hook.) Ching. There has been much debate on the systematic position of Triblemma, and this genus has always been included in Diplazium. Here, we have described new features of tracheary elements, epidermis, spore, scale, and rachis of Triblemma revealed using light and scanning electron microscopy and proposed that Triblemma is closely related to Athyriopsis, which conflicts with the traditional viewpoint.

Permineralized conifer-like leaves from the Jurassic of Patagonia (Argentina) and its paleoenvironmental implications.

Anatomically preserved conifer-like leaves from the Middle Jurassic La Matilde Formation at the Barda Blanca locality in the Gran Bajo de San Julián area, southern Patagonia are described here. Leaves are assigned to conifers based on the following foliar features: thick-walled epidermal cells, a sclerenchymatic hypodermis, resin canals and transfusion tracheids associated with the vascular bundle. General mesophyll anatomy and inferred foliar morphology suggest a similarity to large, broad, linear-lanceolate, multi-veined conifer-like leaves. The general foliar habit indicates an affinity with the large, multi-veined leaves of the Araucariaceae; especially with those exhibited by the species of the Araucaria sections, Araucaria and Bunya. Anatomically, the permineralized leaves exhibit xeromorphic foliar features, including thick-walled epidermal cells, an isobilateral mesophyll with well-developed palisade cells and mechanical tissue. The general leaf anatomy shown by the Patagonian specimens along with sedimentological data may suggest that during the deposition of the La Matilde Formation at the Barda Blanca locality, the parent plant was well adapted to the environmental conditions, which probably consisted of a high light intensity with an adequate quantity of water in the soil, which increased the maximum leaf conductance of CO2.

Anatomical structures of fine roots of 91 vascular plant species from four groups in a temperate forest in Northeast China.

Fine roots of plants play an important role in terrestrial ecosystems. There is a close association between the anatomical characteristics and physiological and ecological functions of plants, but we still have a very limited knowledge of anatomical traits. For example, (1) we do not know if herbs and grasses have anatomical patterns similar to those of woody plants, and (2) the variation among different woody plants in the same ecosystem is unclear. In the present study, we analysed the anatomical structures of the fine root systems of various groups of vascular plants (ferns, eudicot herbs, monocots and woody plants) from the same ecosystem (a natural secondary forest on Mao'er Mountain, Heilongjiang, China) to answer the following questions: (1) How does the anatomy of the fine roots change with root order in various plant groups in the same ecosystem? (2) What is the pattern of variation within group? The results show that anatomical traits can be divided into 3 categories: traits that indicate the root capacity to transport resource along the root (stele diameter, xylem cell diameter and xylem cell area); traits that indicate absorptive capacity cortical thickness, (the number of cortical cell layers and the diameter of cortical cells); and traits that are integrated indicators (diameter and the stele to root diameter ratio). The traits indicate the root capacity to transport resource along the root order is generally similar among groups, but absorptive capacity is very different. The shift in function is the main factor influencing the fine root anatomy. Some traits show large variation within groups, but the variations in other traits are small. The traits indicate that the lower-order roots (absorbing roots) in distinct groups are of the first one or two root order in ferns, the first two or three orders in eudicot herbs, the first (only two root orders) or first two orders (more than three root orders) in monocots and the first four or five root orders in woody plants and the other roots are higher-order roots (transport roots). The result will helpful to understand the similarities and differences among groups and the physiological and ecological functions of plant roots.

Functional diversity and convergence in the evolution of plant reproductive structures.

Background and Aims: Structures that simultaneously perform many functional roles are likely to show a variety of morphological solutions to these demands, and thus probably exhibit high morphological disparity. In contrast, specialization for a few simple functions should result in a more limited suite of morphologies. We explore this idea using lycopsid reproductive structures, which, throughout their history, have performed a limited set of functional roles compared with the reproductive structures of other plant groups such as seed plants. Methods: We scored living and fossil lycopsid taxa for 18 discrete character measurements and several continuous traits, including sporangium size, supporting axis diameter, and strobilus length and width. We used the discrete characters to construct a multivariate morphospace for lycopsid reproductive morphology through time, and the continuous characters to test whether fossil and extant lycopsids show similar patterns of tissue allocation within reproductive structures. Results: Lycopsids occupy similar areas of reproductive morphospace and show similar patterns of tissue allocation over most of their history, alternating between diffuse fertile zones with leaf-like sporophylls and compact strobili with specialized sporophylls that allow sporangia to be closely packed while also protected during their development. Growth habit also plays an important role in lycopsid reproductive evolution, broadly influencing the size and shape of reproductive structures. Conclusions: Lycopsid reproductive structures are primarily specialized for densely packaging sporangia, and are consistent with the idea that performing limited functional roles is associated with reduced morphological disparity. Morphologies similar to lycopsid strobili are also found in other groups with simple, wind-dispersed propagules, suggesting that the same processes occur across plant lineages.

A revised classification of Chinese Davalliaceae based on new evidence from molecular phylogenetics and morphological characteristics.

Although the phylogenetic framework of Davalliaceae is known, the classification of Chinese Davalliaceae is still controversial. In this study, a molecular phylogenetic tree of 60 accessions, including 29 species produced in China, was constructed using five plastid DNA markers-atpB, atpB-rbcL, rbcL, rbcL-accD, and accD. New data on studied specimens, field investigations, and scanning electron microscopy analysis of leaf epidermis and spores were used to reclassify Chinese Davalliaceae. The taxonomic position of Davallia canariensis was confirmed based on new evidence and a new key to sections of Chinese Davalliaceae was proposed. The taxonomically controversial genus Paradavallodes was confirmed as a polyphyletic group, and it was assigned to Davallia sect. Trogostolon and Davallia sect. Davallodes. Further, species endemic to China were delimited, 21 species were admitted to six sections of Davallia, two new combinations were proposed, two new synonyms were defined and a new key to Chinese species of Davalliaceae was presented.

Macrofossil evidence unveiling evolution of male cones in Ephedraceae (Gnetidae).

BACKGROUND: Male cones of modern Ephedraceae are compound and compact. No fossil evidence has so far been found to support an origin of the compact compound male cone from a hypothetical loosely-arranged shoot system. RESULTS: Here we describe a new macrofossil taxon, Eamesia chinensis Yang, Lin, Ferguson et Wang, gen. et sp. nov., from the Early Cretaceous of western Liaoning, northeastern China. It was an ephedroid shrub bearing male spikes terminal to twigs, but differs from modern Ephedraceae by its loosely-arranged male cones, the axillary male shoot consisting of an elongated synangiophore on which leaf-like foliar organs were inserted, and four sessile synangia terminal to the apex. CONCLUSIONS: The morphology of this fossil suggests that the modern compact male cone of Ephedra was indeed derived from a once loosely-arranged shoot system, and the male reproductive unit originated from a once elongated axillary male shoot. This new fossil species thus provides a transitional link from the hypothetical ancestral shoot system to the modern compact morphology. Changes of habitat from closed humid forests to open dry deserts and shifts of the pollination syndrome may have acted as the driving forces behind this morphological evolution.