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.     

<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.     

Comparative analysis of complete chloroplast genome sequences of two subtropical trees, <Emphasis Type="Italic">Phoebe sheareri</Emphasis> and <Emphasis Type="Italic">Phoebe omeiensis</Emphasis> (Lauraceae)

Phoebe is an economically important genus from the family Lauraceae. It is widely distributed in tropical and subtropical Asia, but systematics of the genus is unclear, and currently there is no species-level phylogeny. Here, we determined the complete chloroplast genome sequences of two species with long-range PCR and next genome sequencing technologies, and identified mutation sites and highly variable regions. These highly variable sites were used to reconstruct the phylogeny. The plastomes of Phoebe sheareri and P. omeiensis were 152, 876, and 152, 855 bp, respectively. Comparative genomic analysis indicated that there are 222 mutation sites including 146 substitutions, 73 indels, and 3 microinversions in both plastomes. Fifty-six single-nucleotide changes were identified in gene-coding regions, and 45 microsatellite sites were found for use in species identification. Fourteen divergence hotspots of 38 variable regions were located. Phylogeny was reconstructed using a Bayesian and maximum likelihood approach for 12 Phoebe species and other five related Lauraceae based on 15 of the highly variable regions including accD-psaI, atpB-rbcL, ndhC-trnV, ndhF-rpl32, petA-psbJ, psaA, psbA-trnH, rbcL, rps8-rpl14, rps16-trnQ, rpl32-trnL, trnC-petN, trnL-trnF, trnS-trnG, and ycf1 indicated that variability in the chloroplast regions proposed as variable is enough to detect divergence events among 12 taxa of Phoebe, and that maybe also useful to help to elucidate further relationships among other taxa of the genus.     

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.     

New Zealand Eocene,Oligocene and Miocene Macrofossil and Pollen Records and Modern Plant Distributions in the Southern Hemisphere

The modern New Zealand flora has a relatively low number of families and genera in relation to land area, but well-preserved macrofossils and pollen from three sites in southern New Zealand suggest that the floras in Eocene, Oligocene and Miocene times were much more diverse at the generic level. At Pikopiko, Southland, a late Eocene in situ forest with fern understory was dominated by conifers, Casuarinaceae, Lauraceae, Nothofagus, Proteaceae, and mesothermal angiosperms including palms (aff. Calamus), Sapindaceae: Cupaniae and Picrodendraceae. At Newvale Mine, Southland, a leaf bed within a thick lignite seam represents leaf fossils preserved in a late Oligocene oligotrophic bog. This site demonstrates that Agathis, Dacrycarpus, Dacrydium, Halocarpus, Microcachrys, Podocarpus and Phyllocladus coexisted with diverse angiosperms including Nothofagus, Gymnostoma, Cunoniaceae, Ericaceae, Sapindaceae and several Proteaceae. Pollen data add Meliaceae, Myrtaceae, Onagraceae and Rubiaceae to the flora. At Foulden Maar, Otago, mummified leaves and flowers, including several with in situ pollen, demonstrate the existence of a diverse flora surrounding an Early Miocene lake. This site contains numerous monocot macrofossils including Astelia, Cordyline, Ripogonum and Typha, as well as the oldest fossils known for Orchidaceae and Luzuriagaceae. This flora was dominated by Lauraceae with affinities to Cryptocarya and Litsea, but other families include Araliaceae, Cunoniaceae, Elaeocarpaceae, Euphorbiaceae sensu lato, Menispermaceae, Myrsinaceae, Myrtaceae, Onagraceae, Proteaceae and Sterculiaceae. Many ferns, conifers, and Nothofagus are from lineages with Gondwanan ancestors, whereas other taxa show links to Australia (e.g., Gyrostemonaceae), New Caledonia (e.g., Beauprea) and South America (e.g., Luzuriaga, Fuchsia). Many of these taxa are now extinct in New Zealand, and therefore indicate much wider biogeographic ranges for many families and genera in the past.     

Taxonomic revisions in the Microstromatales: two new yeast species,two new genera,and validation of <Emphasis Type="Italic">Jaminaea</Emphasis> and two <Emphasis Type="Italic">Sympodiomycopsis</Emphasis> species

Environmental sampling yielded two yeast species belonging to Microstromatales (Exobasidiomycetes, Ustilaginomycotina). The first species was collected from a leaf phylloplane infected by the rust fungus Coleosporium plumeriae, and represents a new species in the genus Jaminaea, for which the name Jaminaea rosea sp. nov. is proposed. The second species was isolated from air on 50% glucose media and is most similar to Microstroma phylloplanum. However, our phylogenetic analyses reveal that species currently placed in Microstroma are not monophyletic, and M. phylloplanum, M. juglandis and M. albiziae are not related to the type species of this genus, M. album. Thus, Pseudomicrostroma gen. nov. is proposed to accommodate the following species: P. glucosiphilum sp. nov., P. phylloplanum comb. nov. and P. juglandis comb. nov. We also propose Parajaminaea gen. nov. to accommodate P. albizii comb. nov. and P. phylloscopi sp. nov. based on phylogenetic analyses that show these are not congeneric with Jaminaea or Microstroma. In addition, we validate the genus Jaminaea, its respective species and two species of Sympodiomycopsis and provide a new combination, Microstroma bacarum comb. nov., for the anamorphic yeast Rhodotorula bacarum. Our results illustrate non-monophyly of Quambalariaceae and Microstromataceae as currently circumscribed. Taxonomy of Microstroma and the Microstromataceae is reviewed and discussed. Finally, analyses of all available small subunit rDNA sequences for Jaminaea species show that J. angkorensis is the only known species that possess a group I intron in this locus, once considered a potential feature indicating the basal placement of this genus in Microstromatales.     

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).     

Anatomy and photosystem II activity of <Emphasis Type="Italic">in vitro</Emphasis> grown <Emphasis Type="Italic">Aechmea blanchetiana</Emphasis> as affected by 1-naphthaleneacetic acid

Auxins are one of the main regulators of in vitro plant growth and development. However, the mechanisms, by which auxins, such as 1-naphthaleneacetic acid (NAA), affect in vitro root and leaf anatomy and photosystem function, remain unclear. Accordingly, the aim of the present study was to analyze the effect of different NAA concentrations on the anatomy and photosynthetic performance of in vitro-propagated Aechmea blanchetiana and to determine whether such a treatment affects micropropagated plants after acclimatization. In vitro-established A. blanchetiana plants were transferred to culture media that contained 0, 2, 4, or 6 μM NAA, and after 50 d, they were transplanted into plastic seedling trays with a commercial substrate and cultivated for 60 d in a greenhouse. The plants were evaluated after a 50-d in vitro NAA exposure (growth traits, chlorophyll α fluorescence, and root and leaf anatomy) and after 60 d of acclimatization in the greenhouse (root and leaf growth). Changes induced by NAA in root anatomy might improve uptake of minerals and sugars from the medium, thereby increasing the in vitro growth. In the leaves, the lowest chlorenchyma thickness and sclerenchyma area were observed in plants grown without NAA, and NAA exposure also improved photosystem II activity. The highest ex vitro growth rate was observed for plants that were propagated with 4 μM NAA. Therefore, the use of NAA during in vitro propagation can improve the anatomical and physiological quality of A. blanchetiana plants, as well as to improve ex vitro transfer.     

Revision of species previously reported from Brazil under <Emphasis Type="Italic">Morganella</Emphasis>

In our study, seventy specimens from Brazilian herbaria, as well as from NY and PDD, identified as Morganella, were analyzed. They included the paratype of M. mexicana and holotypes of M. arenicola, M. albostipitata, M. compacta, M. nuda, M. rimosa. Specimens were studied morphologically and used for DNA extraction and amplification of ITS and LSU nuclear ribosomal DNA. New sequences were obtained and compared with homologous sequences from GenBank. As a result of these analyses, a new species and six new combinations are proposed. Morphological review and discussion are provided for the species from Brazil.     

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.     

Interspecific variation in extracellular polysaccharide content and colony formation of <Emphasis Type="Italic">Microcystis</Emphasis> spp. cultured under different light intensities and temperatures

Single cells of five different Microcystis species (M. ichthyoblabe, M. viridis, M. flos-aquae, M. wesenbergii, and M. aeruginosa) were batch-cultured at different temperatures and light intensities: (a) 25 °C and 50 μmol photons m^?2 s^?1 (control culture); (b) 25 °C and 10 μmol photons m^?2 s^?1; and (c) 15 °C and 50 μmol photons m^?2 s^?1. The extracellular polysaccharide content was significantly higher in treatments b and c than in the control treatment. All Microcystis species existed as single cells under the control treatment but formed colonies in treatments b and c. All of the colonies were irregular with indistinct margins. M. ichthyoblabe, M. viridis, M. flos-aquae, and M. wesenbergii formed colonies with similar morphologies and their cells were loosely aggregated. In contrast, M. aeruginosa formed denser colonies with no distinct holes. The colony morphologies differed from the classic morphology of M. ichthyoblabe field-grown colonies but resembled that of small colonies found in Lake Taihu (Yangtze Delta Plain, China) during early spring. This indicates that field- and laboratory-grown colonies are governed by similar formation processes. We suggest that in laboratory and field environments, M. ichthyoblabe (or M. flos-aquae) colonies are representative of small colonies formed from single Microcystis cells, whereas the morphology of older colonies evolves to resemble M. wesenbergii and M. aeruginosa colonies.     

Leaf anatomy of <Emphasis Type="Italic">Acrocomia</Emphasis> (Arecaceae): an additional contribution to the taxonomic resolution of a genus with great economic potential

Acrocomia is one of the most complex genera to understand in the Neotropical Arecaceae, and there is no consensus on the number of species in the genus. A comparative study of leaf anatomy was conducted on seven species of Acrocomia: one with a wide distribution in the Americas (A. aculeata), five endemic to different regions of Brazil, Bolivia and Paraguay (A. emensis, A. glaucescens, A. hassleri, A. intumescens and A. totai) and one endemic to Cuba (A. crispa). Characters that unify the species of Acrocomia include the following: epidermis covered with cuticle, hypodermis on both sides of the leaflets, non-vascular fiber bundles, and primary and secondary vascular bundles. The shape of the leaflet margin, the distribution of the primary vascular bundles and fiber bundles, the number of idioblasts with raphides and the size of the vascular bundles, along with other characteristics, were used to distinguish these species in Acrocomia.     

Phylogenetic relationship and time of divergence of <Emphasis Type="Italic">Mus terricolor</Emphasis> with reference to other <Emphasis Type="Italic">Mus</Emphasis> species

Mitochondrial DNA control region of Mus terricolor, three aboriginal species M. spretus, M. macedonicus, M. spicilegus; the Asian lineage M. caroli, M. cervicolor, M. cookii; and the two house mice, M. musculus domesticus and M. m. castaneus were analysed to estimate the substitution rate, phylogenetic relationship and the probable time of divergence. Results showed that M. spretus, M. caroli and M. terricolor are highly diverged from each other (caroli/terricolor = 0.146, caroli/spretus = 0.147 and terricolor/spretus = 0.122), whereas M. spretus showed less divergence with two house mice species (0.070 and 0.071). Sequence divergence between M. terricolor and the Palearctic group were found to be ranging from 0.121 to 0.134. Phylogenetic analysis by minimum evolution, neighbour-joining, unweighed pair group method with arithmetic mean and maximum parsimony showed almost similar topology. Two major clusters were found, one included the Asian lineage, M. caroli, M. cookii and M. cervicolor and the other included the house mice M. m. domesticus, M. m. castaneus and the aboriginal mice M. macedonicus and M. spicilegus along with M. spretus, forming the Palearctic clade. M. terricolor was positioned between the Palearctic and Asian clades. Results showed that Palearctic-terricolor and the Asian lineages diverged 5.47 million years ago (Mya), while M. terricolor had split around 4.63 Mya from their ancestor. M. cervicolor, M. cookii and M. caroli diverged between 4.70 and 3.36 Mya, which indicates that M. terricolor and the Asian lineages evolved simultaneously. M. spretus is expected to have diverged nearly 2.9 Mya from their most recent common ancestor.     

<Emphasis Type="Italic">Metarhizium bibionidarum</Emphasis> and <Emphasis Type="Italic">M. purpureogenum</Emphasis>: new species from Japan

Two new species of Metarhizium, M. bibionidarum and M. purpureogenum are described from Japan. Metarhizium bibionidarum is the phylogenetic sister species of M. pemphigi and a member of the M. flavoviride species complex. It is distinguished morphologically from M. pemphigi by its larger conidia. The species is based on a collection of an infected March fly larva (Diptera: Bibionidae) but is also known to occur on fruit beetle (Coleoptera: Scarabaeidae) encountered in France. Metarhizium purpureogenum was isolated from soil by plating and insect baiting methods and represents a unique phylogenetic lineage placed outside the M. anisopliae and M. flavoviride species complexes. Three isolates of M. purpureogenum excreted a distinctive red-purple pigment into agar medium when co-cultured with M. robertsii or Aspergillus oryzae.     

<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.     

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.     

Foliar water uptake,a widespread phenomenon in temperate woodland ferns?

Most woodland ferns thrive under conditions of high air humidity, frequent precipitation and exposure to extended periods of leaf wetness, but it is not known how widespread foliar water uptake is in this plant group. In a tracer experiment with deuterated water (²H₂O) applied to the leaf surface of five temperate woodland ferns (Athyrium filix-femina, Dryopteris filix-mas, Polystichum aculeatum, Polystichum braunii and Asplenium scolopendrium), we tested (1) whether these species exhibit foliar water uptake and (2) whether the capability to absorb water through the leaf epidermis increases with the frequency of epidermal trichomes. All species had significantly higher abundances of ²H in tissue water, when extracted distant to the place of application, compared to the background level (0.052–0.504 vs. 0.015 at.%), evidencing uptake through the epidermis and leaf-internal translocation. A positive relation between trichome density and ²H incorporation was found only for the second-order pinnae but not for the more central frond sections. The results suggest that foliar water uptake may be widespread among temperate woodland ferns across different families and that leaf trichome structure probably influences this process.