Achene morphology of Saussurea species (Asteraceae,Cardueae) in Korea and its systematic implications

Achene size and shape, surface sculpturing, and pericarp and testa wall structure of 23 Korean Saussurea spp. were investigated using scanning electron microscopy (SEM) and light microscopy to evaluate the infrageneric relationships and assess their systematic significance. Achene size categories and thickness of the testa epidermis were distinguished using biometric measurements. Four basic types of surface pattern were observed: (1) lineate; (2) striate; (3) reticulate; and (4) colliculate. Saussurea rorinsanensis was found to have some unique achene characteristics, such as a fusiform achene, uniform pappus, presence of epidermal hairs and tangentially elongated, narrow testa epidermal cells. The characteristic achene features for species were found to be achene size and shape, hilum position, surface sculpture, pappus composition, morphology of the pericarp wall and thickness of the testa epidermis. Based on 16 morphological and achene characters, a cladistic analysis resolved three well‐supported clades, with S. eriophylla as the first‐branching taxon. Saussurea pulchella and S. japonica, both belonging to Saussurea subgenus Theodorea, were distant from each other in the 50% majority rule consensus tree and the character distribution cladogram. This cladistic analysis of achene morphology and anatomy should be regarded as giving us a tentative picture of the phylogenetics of Saussurea, and this study may serve as a reference for future hypotheses and studies on the characterization and classification of Saussurea spp. in Korea.     

External water transport is more important than vascular transport in the extreme atmospheric epiphyte Tillandsia usneoides (Spanish moss)

Most epiphytic bromeliads, especially those in the genus Tillandsia, lack functional roots and rely on the absorption of water and nutrients by large, multicellular trichomes on the epidermal surfaces of leaves and stems. Another important function of these structures is the spread of water over the epidermal surface by capillary action between trichome “wings” and epidermal surface. Although critical for the ultimate absorption by these plants, understanding of this function of trichomes is primarily based on light microscope observations. To better understand this phenomenon, the distribution of water was followed by its attenuation of cold neutrons following application of H₂O to the cut end of Tillandsia usneoides shoots. Experiments confirmed the spread of added water on the external surfaces of this “atmospheric” epiphyte. In a morphologically and physiologically similar plant lacking epidermal trichomes, water added to the cut end of a shoot clearly moved via its internal xylem and not on its epidermis. Thus, in T. usneoides, water moves primarily by capillarity among the overlapping trichomes forming a dense indumentum on shoot surfaces, while internal vascular water movement is less likely. T. usneoides, occupying xeric microhabitats, benefits from reduction of water losses by low‐shoot xylem hydraulic conductivities.     

Effects of Salicylic Acid on the Structure of Second Leaves of Hordeum Vulgare L.

The structure and ultrastructure of the second leaves of 10-d-old barley plants (Hordeum vulgare L. cv. Alfa) was investigated after long-term treatment with salicylic acid (SA) in concentrations of 0.1, 0.5 and 1.0 mM. The treatment induced: 1) suppressed bulliform cells formation in the adaxial epidermis (1.0 mM SA); 2) reduction of apoplast in the mesophyll (0.5 and 1.0 mM SA); 3) formation of invaginations (0.1 and 0.5 mM SA) and proliferations (0.5 mM SA); and 4) thylakoid destruction and coagulation of the stroma (1.0 mM SA).     

Epidermal stem cells: interactions in developmental environments

Homeostasis of continuously renewing adult tissues, such as the epidermis of the skin, is maintained by epidermal stem cells (EpiSC), which are a small population of undifferentiated, self-renewing basal keratinocyte cells that produce daughter transit amplifying (TA) cells to make up the majority of the proliferative basal cell population in the epidermis. We have isolated EpiSC from neonatal and adult skin, and shown that these cells can regenerate an epidermis that lasts long term in vitro and in vivo, and that permanently expresses a recombinant gene in the regenerated tissue (Bickenbach and Dunnwald, 2000; Dunnwald et al., 2001). When we injected murine EpiSC into the developing blastocyst environment of the mouse, we found that both neonatal and adult EpiSC retained some ability to participate in the formation of tissues from all three germ layers (Liang and Bickenbach, 2002; Bickenbach and Chinnathambi, 2004; Liang et al., 2004). Although it appears evident that EpiSC act as pluripotent stem cells, how this reprogramming takes place is not understood. EpiSC might directly transdifferentiate into other cell types or they might first dedifferentiate into a more primitive cell type, and then proceed to develop along a cell lineage pathway. To begin to unravel this, we co-cultured EpiSC with embryonic stem (ES) cells, and found that EpiSC could alter their cell lineage protein expression to that of a more primitive cell type. We also placed EpiSC in a wounded environment and found that EpiSC interacted with the mesenchymal cells repopulating the wound bed. Our findings indicate that the population of cells that we isolate as EpiSC has a pluripotent capability. This has led us to postulate a paradigm shift for somatic stem cells. We propose that tissues maintain a sequestered population of uncommitted stem cells that retain a regenerative response which is enhanced when the cells are exposed to developmental or stress influences.     

Silica deposition in relation to ageing of leaf tissues in Sasa veitchii (Carriere) Rehder (Poaceae: Bambusoideae)

BACKGROUND AND AIMS: Silica deposition is one of the important characteristics of plants in the family Poaceae. There have been many investigations into the distribution, deposition and physiological functions of silica in this family. Two hypotheses on silica deposition have been proposed based on these studies. First, that silica deposition occurs passively as a result of water uptake by plants, and second, that silica deposition is controlled positively by plants. To test these two apparently contradictory hypotheses, silica deposition in relation to the ageing of leaf tissues in Sasa veitchii was investigated. METHODS: Tissues were examined using a light microscope and a scanning electron microscope equipped with an energy dispersive X-ray microanalyser. KEY RESULTS: The deposition process differed depending on cell type. In mesophyll tissue, fusoid cells deposited large amounts of silica depending on leaf age after maturation, while chlorenchyma cells deposited little. In epidermal tissue, comprised of eight cell types, only silica cells deposited large amounts of silica during the leaf's developmental process and none after maturation. Bulliform cells, micro-hairs and prickle hairs deposited silica densely and continuously after leaf maturation. Cork cells, guard cells, long cells and subsidiary cells deposited silica at low levels. CONCLUSIONS: The significance of these observations is discussed in relation to the two hypotheses proposed for silica deposition in Poaceae. The results of the present study clearly indicate that both hypotheses are compatible with each other dependent on cell types.     

Vitamin D regulated keratinocyte differentiation

The epidermis is the largest organ in the body. It is comprised primarily of keratinocytes which are arranged in layers that recapitulates their programmed life cycle. Proliferating keratinocytes are on the bottom-the stratum basale. As keratinocytes leave the stratum basale they begin to differentiate, culminating in the enucleated stratum corneum which has the major role of permeability barrier. Calcium and the active metabolite of vitamin D, 1,25(OH)(2)D(3), play important roles in this differentiation process. The epidermis has a gradient of calcium with lowest concentrations in the stratum basale, and highest concentrations in the stratum granulosum where proteins critical for barrier function are produced. Vitamin D is made in different layers of the epidermis, but 1,25(OH)(2)D(3) is made primarily in the stratum basale. Together calcium and 1,25(OH)(2)D(3) regulate the ordered differentiation process by the sequential turning on and off the genes producing the elements required for differentiation as well as activating those enzymes involved in differentiation. Animal models in which the sensing mechanism for calcium, the receptor for 1,25(OH)(2)D(3), or the enzyme producing 1,25(OH)(2)D(3) have been rendered inoperative demonstrate the importance of these mechanisms for the differentiation process, although each animal model has its own phenotype. This review will examine the mechanisms by which calcium and 1,25(OH)(2)D(3) interact to control epidermal differentiation.     

A keratin K5Cre transgenic line appropriate for tissue-specific or generalized Cre-mediated recombination

We describe here a mouse line bearing a bovine keratin K5Cre recombinase transgene. These mice showed a dual pattern of Cre-mediated recombination, depending on the parent transmitting the transgene. In paternal transmission, recombination occurred specifically in the skin and stratified epithelia-as expected according to the expression of endogenous keratin K5. However, constitutive recombination between loxP sites transmitted by the sperm took place when the mother possessed the K5Cre transgene, even when the transgene was absent in the progeny. Cre expression in late-stage oocytes, with the Cre protein persisting into the developing embryo, leads to the constitutive recombination observed. Thus, this transgenic line allows for both tissue-specific and generalized recombination, depending on the breeding scheme.     

Observations on the histochemistry and ultrastructure of regenerating caudal epidermis of the tuatara Sphenodon punctatus (Sphenodontida,Lepidosauria, Reptilia)

Study of the histology, histochemistry, and fine structure of caudal epidermal regeneration in Sphenodon punctatus through restoration of a scaled form reveals that the processes involved resemble those known in lizards. Following establishment of a wound epithelium (WE), subjacent scale neogenesis involves epidermal downgrowths into the dermis. Although the process is extremely slow, and most new scales do not overlap, their epidermal coverings reestablish epidermal generation (EG) formation. As in lizards, the flat, alpha-keratogenic, WE cells contain lipids as revealed by their affinity for Sudan III. A few mucous cells that store large PAS-positive mucus-like granules also occur in WE. During differentiation of WE cells, among the bundles of 70-nm tonofilaments are many lamellar bodies (LBs) and mucous granules (MGs) that discharge their contents into the cytoplasm and extracellular spaces producing a strongly PAS-positive keratinized tissue. Richness of epidermal lipids coexistent with mucus is a primitive characteristic for amniote vertebrates, probably related to functions as a barrier to cutaneous water loss (CWL). As scale neogenesis begins, beneath the superficial WE appear 3-5 layers of irregularly shaped cells. These contain tonofilament bundles surrounded by small, round keratohyalin-like granules (KHLGs) and a keratinized matrix with beta-keratin packets and a 3-5-nm thick keratin granulation. This mixture of alpha- and beta-keratogenic capacities resembles that seen in the innermost cells of a normal tuatara epidermal generation. As in the latter, but in contrast to both normal and regenerating lizard epidermis, no definable shedding complex with interdigitating clear layer and oberhautchen cells occurs (Alibardi and Maderson, 2003). The tortuous boundaries, and merging beta-keratin packets, identify subjacent keratinizing cells as precursors of the typical stratified, squamous beta-layer seen in long-term regenerated caudal skin wherein the entire vertical sequence of epidermal layers resembles that of normal scales. The sequence of events in caudal epidermal regeneration in S. punctatus resembles that documented for lizards. Observed differences between posttrauma scale neogenesis and scale embryogenesis are responses to functional problems involved in, respectively, restoring, or forming, a barrier to CWL while accommodating rapid somatic growth.     

Structure of the Photosynthetic Apparatus in Leaves of Freshwater Hydrophytes: 2. Quantitative Characterization of Leaf Mesophyll and the Functional Activity of Leaves with Different Degrees of Submersion

The structure of leaf photosynthetic elements was investigated on 42 boreal plant species characterized by different degrees of submergence (helophytes, neustophytes, and hydatophytes). Six main types of mesophyll structures were identified. Quantitative characteristics for the mesostructure of the photosynthetic apparatus in these groups were determined, such as the size and abundance of cells and chloroplasts in the mesophyll and epidermis, the number of plastids per cell in each tissue, the total surface area of the mesophyll cells, epidermal cells, and chloroplasts per unit leaf area. Analysis showed that quantitative characteristics of the photosynthetic apparatus in hydrophytes are determined by two factors: (a) the degree of leaf submergence and (b) the type of mesophyll structure. With an increasing degree of immersion in water, the mesophyll types change in a sequence isopalisade dorsoventral homogeneous. The leaves become thinner, their weight per unit area diminishes, cells and chloroplasts become less numerous (on a per unit leaf area basis), but their dimensions become larger. Adaptation to aquatic medium is also manifested in the increasing contribution of the epidermal tissue to the overall photosynthesis: in submerged leaves, the epidermis accounts for more than 50% of the photosynthetic activity. The occurrence of six structural types of leaves contrasting in their characteristics was confirmed by discriminatory analysis according to the qualitative parameters of mesophyll.