Comparative wood anatomy of the Cupressaceae and correspondence with phylogeny,with special reference to the monotypic taxa

Wood anatomy is one of the tools used for taxonomic classification of species. By combining this tool with molecular phylogeny, the current groupings made in morphological studies can be discussed. This study describes the wood anatomy of the monotypic genera of Cupressaceae and analyses the features that could represent synapomorphies of the principal clades recovered by molecular phylogeny. The wood anatomical study shows the high homogeneity of this family, revealing the presence of common ancestral features that support the union between Taxodiaceae and Cupressaceae s.s. and the separation of Sciadopitys. It also supports the group formed by Taxodiaceae in Cupressaceae s.l. No clear division was observed between the callitroid and cupressoid clades. Some wood anatomical differences were observed in the Fitzroya–Diselma–Pilgerodendron and Microbiota–Platycladus–Tetraclinis associations. The wood anatomical features common to the family, such as axial tracheids without helical thickenings, homogeneous rays, cupressoid cross-field pits and the presence of a warty layer, are put forward as possible synapomorphies for Cupressaceae s.l. The clade-specific synapomorphies are taxodioid cross-field pits for taxodioid and sequoioid clades, absence of a well-defined torus in Thuja–Thujopsis and torus extensions in Diselma–Fitzroya–Widdringtonia.     

Systematic and phylogenetic implications of the wood anatomy of six Neotropical genera of Primulaceae

Our main goals were to identify diagnostic characters at the species, genus, and subfamily levels, find anatomical features with potential for future morphological and molecular (combined) phylogenetic analyses, and to reconstruct the evolution of wood anatomical characters in two subfamilies of Primulaceae in a molecular phylogenetic framework. We investigated twenty-seven species from the woody Myrsinoideae (4 genera) and Theophrastoideae (2 genera) using scanning electron, light, and epifluorescence microscopy. Samples were prepared using standard protocols. Based on the wood anatomical characters, we were able to identify synapomorphies and to detect evolutionary trends of interest for the genera and subfamilies. Both subfamilies share the presence of diffuse porosity, simple perforation plates, septate fibres, and scanty paratracheal axial parenchyma. Theophrastoideae species have rays?>?10 cells wide and short (<?350 µm) vessel elements, and Myrsinoideae have breakdown areas in rays and longer vessel elements. Ardisia and Stylogyne have scalariform intervessel pits, Myrsine exhibit breakdown areas in rays, and two Cybianthus species from subgenus Weilgetia have distinguishing features (e.g., scalariform perforation plate in C. nemoralis and the absence of rays in C. densiflorus). Overall, when combining characters, we were able to segregate the Neotropical Primulaceae subfamilies and genera from each other and from the subfamily Maesoideae based on wood anatomy.     

Stem anatomy of neotropical <Emphasis Type="Italic">Dioscorea</Emphasis> L. (Dioscoreaceae) and its importance to the systematics of the genus

We anatomically characterized the aerial stems of the genus Dioscorea and evaluated the use of these anatomical characteristics to better understand the taxonomy, systematics, and diversity of the component species in the neotropics. Fragments of the aerial stems of 23 species were collected for anatomical analyses utilizing conventional cytohistological techniques. Stem anatomy did not support the subgenera and neotropical sections of Dioscorea, although D. sect. Monadelpha demonstrated similar anatomical characteristics with the representatives analyzed in the present work, supporting the possible monophyletism of that section. Some New World species demonstrated characteristics described earlier for Old World species, such as two phloem units internal to the large-diameter vessels. New characters to the vascularization of new world species are described, as a single ring of vascular bundles, intercalating common and cauline bundles. Tylosoids and actinocytic stomata were the first time reported in Dioscorea, and the vascular patterns are revisited with a systematic approach.     

The contribution of foliar micromorphology and anatomy to the circumscription of species within the <Emphasis Type="Italic">Chusquea ramosissima</Emphasis> informal group (Poaceae,Bambusoideae, Bambuseae)

Chusquea is a diverse but monophyletic genus of Neotropical woody bamboos from primarily montane forests that comprises four well-supported lineages: subg. Magnifoliae, subg. Platonia, subg. Rettbergia, and the Euchusquea clade (comprising subg. Swallenochloa and subg. Chusquea). However, the relationships among clades or taxa within the Euchusquea clade inferred from molecular data are mostly not congruent with those inferred from morphological evidence, consequently limiting our ability to understand species relationships. In this study we generated foliar micromorphological and anatomical data for the Chusquea ramosissima informal group (Chusquea ramosissima, C. tenella, and C. tenuiglumis), and for the putative new species from Bolivia in this group, in order to test the value of these types of data for defining species and to seek potential synapomorphies for this group. Our results demonstrate that epidermal features, mainly with regard to the stomatal apparatus, proved to be more valuable in distinguishing species than anatomical characters. The presence of horizontally elongated silica cells over the veins and adaxial arm cells with invaginations from the abaxial side was shared by all the studied species but is not unique to this group. The type of trichomes, shape of silica bodies, type of arm cells, and midrib structure may be useful to lesser degree. All four species exhibited intraspecific variation in development of the papillae on the long cells. Support for the recognition of the new species from Bolivia is provided by micromorphological characters. An identification key based on leaf blade features is provided for the four studied species.     

A new <Emphasis Type="Italic">Heteropterys</Emphasis> (Malpighiaceae) from semideciduous forest,with notes on wood anatomy

This paper describes and illustrates Heteropterys serrata, a new species endemic to semideciduous forests associated with inselbergs in the state of Espírito Santo, Brazil. Morphological and wood anatomical traits of the new species are compared to those of other species of the Heteropterys Metallophyllis informal group. Based on wood anatomy, H. serrata and H. nitida (the most morphologically similar species) have different axial parenchyma, which is scarce paratracheal in H. nitida (and some other species of the Metallophyllis informal group) and aliform confluent in H. serrata. The most notable morphological and anatomical characters that distinguish the new species are the young hexagonal stems, the unusual widely serrate leaf margins and the aliform confluent axial parenchyma in the wood.     

Comparative leaf micromorphology and anatomy of the dragon tree group of <Emphasis Type="Italic">Dracaena</Emphasis> (Asparagaceae) and their taxonomic implications

Micromorphological features of the leaf epidermis and the inner structure of leaf tissues of eight arborescent taxa of the genus Dracaena were analysed using light and scanning electron microscopy. The plants are xeromorphic or mesomorphic. Their leaves are isobilateral and amphistomatic, and the stomata are anomocytic and tetracytic. The mesophyll in all the species is divided into an outer chlorenchyma and a central region with colourless water-storage cells, chlorophyll cells and vascular bundles. Water-storage cells have wall bands and reticulate thickenings on the walls. The article describes and illustrates several new quantitative and qualitative leaf characters of the dragon tree group. Our findings can be used to identify the dragon tree group leaves, while the shape of epidermal cells and stomata types may be useful in the identification and classification of fragments of fossil leaves. We conclude that D. ombet and D. schizantha are not two distinct species, but should be treated as subspecies of D. ombet. Leaf characters, especially stomata depth on adaxial epidermis, height of adaxial epidermal cells and the presence and thickness of hypodermal fibre bundles markedly differ between geographical groups: Macaronesian species (D. draco and D. tamaranae), the species found in East Africa and Arabian Peninsula (D. ombet subsp. ombet, D. ombet subsp. schizantha, D. serrulata and D. cinnabari) and Southeast Asian species (D. kaweesakii and D. jayniana).     

Anatomical features of species of Cyperaceae from northeastern Brazil

Seventy-three species from 17 genera of Cyperaceae were studied with the aim of identifying and confirming those species with Kranz anatomy. Among the species studied, 36 exhibited Kranz anatomy; 37 did not. Of the four types of Kranz anatomy recognized in Cyperaceae, three were encountered: the chlorocyperoid type in the genera Cyperus, Kyllinga, Lipocarpha, Pycreus, and Remirea; the fimbristyloid type in Abildgaardia, Bulbostylis, and Fimbristylis; and the rhynchosporoid type in Rhynchospora. Non-Kranz anatomy was confirmed in species of the following genera: Becquerelia, Calyptrocarya, Cyperus, Diplacrum, Eleocharis, Fuirena, Hypolytrum, Pleurostachys, Rhynchospora, and Scleria. The anatomical data obtained corroborate earlier studies of species of Cyperaceae as to the presence of Kranz anatomy and the anatomical types in several species and the “Kranzkette” pattern in Cyperus ligularis and Cyperus pohlli.     

Diversity and evolution of leaf anatomical characters in Taxaceae <Emphasis Type="Italic">s.l.—</Emphasis>fluorescence microscopy reveals new delimitating characters

Taxaceae s.l. comprise six genera (including Cephalotaxus) and about 35 species; The present study aims to give new insights into the evolution of this family, especially into the phylogenetic position of Cephalotaxus. Moreover, only little is known about comparative leaf anatomy of this family and this study aims to expose and interpret the diversity and evolution of leaf anatomical characters and to assess their applicability to identify taxa at the generic and species level. A detailed phylogeny was reconstructed, using both maximum likelihood and Bayesian inference, with a combined dataset of four molecular markers from the plastid and nuclear genomes. Leaf sections from 132 specimens, representing 32 species and four varieties (fresh and herbarium material) were inspected, using fluorescence microscopy. Ancestral characters were reconstructed using Mesquite. The phylogenetic analyses provided full support for Cephalotaxus as sister group to Taxaceae s.str. Within the latter, two monophyletic tribes Taxeae (comprising Austrotaxus, Pseudotaxus, and Taxus) and Torreyeae (comprising Amentotaxus and Torreya) were fully supported. Fluorescence microscopy was shown to be very useful for identifying leaf tissues and their constitution. We were able to show that particularly sclerified tissues have highest potential for the discrimination of both freshly collected samples and rehydrated herbarium specimens at the generic and species level. A correlation between the presence of different sclereid types could be shown and sclereids were hypothesized to pose a primitive trait in the evolution of Taxaceae s.l. New identification keys were generated on the basis of leaf anatomical characters. The microscopic method presented here is applicable for further studies within gymnosperms and probably in angiosperms, as well.     

<Emphasis Type="Italic">Compsaster formosus</Emphasis><Emphasis Type="SmallCaps">Worthen &; Miller</Emphasis> (Asteroidea; Echinodermata): A Carboniferous homeomorph of the post-Paleozoic Asteriidae

All adequately known post-Paleozoic asteroids are either assignable to surviving families or closely related families whereas no Paleozoic species assignable to a surviving order has been recognized. The Mississippian speciesCompsaster formosus is similar enough to various Recent taxa in overall form as well as in the form and arrangement of body wall ossicles to raise the issue of affinities: IsC. formosus nested within a phylogenetic branch hitherto known only from post-Paleozoic strata or is it only homeomorphic?The nature of the ambulacral system is critical to interpretation of echinoderms, and post-Paleozoic asteroids share three fundamental ambulacral characters or character suites: dorsal podial pores, staggered positioning of ambulacrals and adambulacrals, and complex articular structures between these two ossicular types.Calliasterella americana, a Carboniferous asteroid, shares the three ambulacral features, although it is distinctive from post-Paleozoic asteroids in other ways.Compsaster formosus exhibits at least two of the three ambulacral characters, although presence of staggering has not been finally established. LikeC. americana, C. formosus differs from post-Paleozoic species in details of ambulacral anatomy as well as aspects of ventral body surface ossicular arrangement. Although approaching crown-group organization,C. formosus nevertheless represents a branch basal to the crown group.Because asteroids are generalists, understanding of life habits ofCompsaster is sketchy in spite of morphological similarities between it and younger genera.Compsaster probably was epifaunal and its overall form is strongly reminiscent of that of Recent predatory asteriids but it is also similar to small-particle feeding echinasterids.     

Phylogenetic Relationships within <Emphasis Type="Italic">Phyllophaga</Emphasis> Harris (sensu <Emphasis Type="Italic">lato</Emphasis>) (Coleoptera: Melolonthidae,Melolonthinae) with Emphasis on <Emphasis Type="Italic">Listrochelus</Emphasis> Blanchard

As partial results of a long-term project for the revision of the supraspecific classification of the American Melolonthini, using phylogenetic methods, interesting information on the relationships between the subgeneric groups of Phyllophaga Harris proposed by Saylor were obtained. The genus Listrochelus was described by Blanchard in 1851. However, in 1940, Saylor reduced it as a subgenus of Phyllophaga. The objective of the present study is to confirm the monophyly of Listrochelus by means of a phylogenetic analysis. A total of 132 species were analyzed; 31 species of them belong to Listrochelus, 76 are from other groups of Phyllophaga, and 25 species are from the outgroup. A morphological matrix with 281 characters was codified. A traditional search with 1000 iterations was performed in TNT. Branch support was investigated using the bootstrap method. Parsimony analysis resulted in ten equally parsimonious trees. The topology obtained in the strict consensus tree shows that the limits of Listrochelus are very clear (bootstrap 99%), supported by five synapomorphies and a combination of eight character states that are not exclusive to the group. Based on the hypothesis obtained, the restitution of the genus Listrochelus is proposed.     

Phylogeny and character evolution of the Neotropical fern genus <Emphasis Type="Italic">Stigmatopteris</Emphasis> (Dryopteridaceae)

We did a phylogenetic analysis on 21 of the 23 species of Stigmatopteris, a neotropical montane genus of wet forest floors. The analysis was based on four plastid markers: rbcL, rps4-trnS, trnG-trnR, and trnL-trnF. Two of the most closely related families to the Dryopteridaceae (Didymochlaenaceae and Hypodematiaceae) and 12 dryopteroid genera were used as outgroups. Eight morphological characters were mapped on the resulting tree. Stigmatopteris was recovered as monophyletic and sister to a clade formed by polystichoid ferns such as Arachniodes, Dryopteris, and Polystichum. Synapomorphies that distinguish Stigmatopteris are 1-pinnate-pinnatifid laminae, visible punctate glands in the mesophyll, the presence of hydathodes, uniseriate cilia on the scale margins, and loss of indusia. Within the genus are two main clades. The first consists of four species characterized by pinnae long-decurrent in at least the distal third of the lamina. The second consists of the remaining 17 species in the analysis and has no known morphological synapomorphies. Nested within this second clade is a subclade of six species endemic to southeastern Brazil. Anastomosing veins, a character often used in keys to distinguish the species, evolved three times within the genus.     

Foliar anatomy of <Emphasis Type="Italic">Beaucarnea</Emphasis> Lemaire (Nolinaceae ss)

Beaucarnea (Nolinaceae) is a Mexican and Guatemalan genus that inhabits dry tropical areas. Most of the species are endangered under the Mexican legislation because they have a high horticultural demand and are threatened by habitat destruction. Species are difficult to recognize, since their differences are a mixture of trunk, inflorescence and fruit characters frequently absent in herbarium specimens. The aim of this work is to describe the foliar anatomy of the 11 species of Beaucarnea in search of characters with taxonomic value to sustain the generic subdivision, and to provide a key for specimens determination based on leaf and growth form. We found that four species (B. compacta, B. gracilis, B. purpusii and B. stricta) have a grooved leaf outline in transverse section, whereas the rest of the species are undulated. Other characters, such as epicuticular waxes, stomatal distribution and density, marginal teeth length and leaf apex provide valuable characters for species determination.     

<Emphasis Type="Italic">In vitro</Emphasis> growth profile and comparative leaf anatomy of the C<Subscript>3</Subscript>–C<Subscript>4</Subscript> intermediate plant <Emphasis Type="Italic">Mollugo nudicaulis</Emphasis> Lam.

Mollugo nudicaulis Lam., commonly known as John’s folly or naked-stem carpetweed, is an ephemeral species of tropical regions. The plant is ideal to study the eco-physiological adaptations of C₃–C₄ intermediate plants. In the present report, in vitro growth profiling of the plant and comparative leaf anatomy under in vitro and ex vitro conditions were studied. In vitro propagation of the plant was carried out on Murashige and Skoog (MS) basal medium augmented with additives and solidified with 0.8% (w/v) agar-agar or 0.16% (w/v) Phytagel?. The concentration of plant growth regulators (PGRs) in the basal medium was optimized for callus induction, callus proliferation, shoot regeneration, and in vitro rooting. The optimum callus induction was obtained from M. nudicaulis seedling hypocotyls. The highest regeneration induction of about 88% or nearly 41 shoots with about 142 leaves per culture vessel was observed from friable callus on MS basal medium solidified with Phytagel? and containing 4.44 μM 6-benzylaminopurine, 4.65 μM kinetin, 2.69 μM naphthaleneacetic acid, and 0.91 μM thidiazuron. In leaf anatomy, differences related to photosynthetic tissue organization were observed in leaves of in vitro and ex vitro plants, which indicated that changes in the environment affected the anatomy of subsequent leaves in plants. This is the first report of an efficient micropropagation protocol for M. nudicaulis, using an indirect organogenesis method. Efforts were made to optimize the concentrations of various PGRs and organic compounds for in vitro growth of regenerated shoots.     

Complex flowers and rare pollinators: Does ant pollination in <Emphasis Type="Italic">Ditassa</Emphasis> show a stable system in Asclepiadoideae (Apocynaceae)?

If on one hand, ant pollination is rare, on the other Asclepiadoideae flowers are the most complex among Eudicots. Both themes are exciting in pollination biology. Although there are records of ants with the ability to remove the pollinarium on this subfamily, the role of these insects as pollinators is not yet known. Therefore, we investigated the interaction between flowers and ants, as well as the associated features in two species of Asclepiadoideae, Ditassa capillaris and D. hastata. The studied species were both visited by Cephalotes inaequalis and Dorymyrmex thoracicus. We analyzed the common traits of ant pollination of the plant species, recorded the number of visit by ants, and carried out experiments of selective pollination, developing an effectivity index. Both Ditassa species have common traits of ant pollination, such as small bright flowers with exposed floral resource. The pollinarium is clip-shaped and resistant to the metapleural gland, which ensures effective pollination. Dorymyrmex thoracicus is a very frequent but not very efficient pollinator, whereas Cephalotes inaequalis is more efficient though at a lower frequency. Therefore, the total contributions of both ants to the pollination of both Ditassa species are similar. The features of both Ditassa species which favor ant pollination, include some synapomorphies within Asclepiadoideae. Furthermore, the efficiency of the ants as pollinators to the reproductive success of the plants was similar despite the different behaviors observed in both ant species. Hence, we suggest that the role of ants in pollination within this subfamily has been neglected.     

<Emphasis Type="Italic">Ramaria chocoënsis</Emphasis> sp. nov., a gomphoid member of <Emphasis Type="Italic">Ramaria</Emphasis> sect. <Emphasis Type="Italic">Dendrocladium</Emphasis> from Colombia,El Chocó, with special regards to rhizomorph anatomy

The new species Ramaria chocoënsis (Gomphales, Basidiomycota) from Colombian Pacific rainforest is described. Despite the gomphoid habit, the new species is inserted in Ramaria sect. Dendrocladium due to rhizomorph anatomy. In rhizomorphs of members of Ramaria sect. Dendrocladium, as in the new species R. chocoënsis, peculiar, thick-walled, gnarled, sphere-bearing hyphae are found, a feature unknown for any other higher fungi. A short survey of hitherto known rhizomorph features within the Gomphales is given. The possibility of putting the members of Ramaria sect. Dendrocladium in a separate genus is taken into account. It would be advisable to have additional data by screening the anatomy of the rhizomorphs in other species, to more accurately predict the classification of Ramaria chocoënsis sp. nov.