Pattern analysis of stem cell differentiation during <Emphasis Type="Italic">in vitro Arabidopsis</Emphasis> organogenesis

Plant somatic cells have the capability to switch their cell fates from differentiated to undifferentiated status under proper culture conditions, which is designated as totipotency. As a result, plant cells can easily regenerate new tissues or organs from a wide variety of explants. However, the mechanism by which plant cells have such remarkable regeneration ability is still largely unknown. In this study, we used a set of meristem-specific marker genes to analyze the patterns of stem cell differentiation in the processes of somatic embryogenesis as well as shoot or root organogenesis in vitro. Our studies furnish preliminary and important information on the patterns of the de novo stem cell differentiation during various types of in vitro organogenesis.     

Adaptive characteristics of the endemic species Satureja horvatii Šilić (Lamiaceae) in mountain-mediterranean and mediterranean habitats

TODOROVIĆ, T. & STEVANOVIĆ, B., 1994. Adaptive characteristics of the endemic species Satureja horvatii Šilić (Lamiaceae) in mountain-mediterranean and mediterranean habitats. Morphoanatomical analysis in two popoulations of Satureja horvatii from habitats of different altitudes in the Orjen-Lovćen mountain massif (Montenegro, Yugoslavia) disclosed important adaptive differences and ecological plasticity of this local endemic species. The Satureja horvatii population growing on Mt. Orjen (1540m) comprises mesoxerophytes, while the population from the foothills of Mt. Lovćen (280 m) is of a typical xerophytic type, as demonstrated by the ecoanatomical study of their leaves.     

Formation of Basal Bodies in Ciliary Epithelium of Molluscs Buccinum undatum L. and Lymnaea stagnalis L.

Electron microscopic studies of the leg ciliary epithelium was carried out in two mollusks. In the epithelium of the leg of adult animals, the centrioles were mostly formed de novo with participation of deuterosomes during the formation of basal bodies. Transformation of the centriolar cylinder in a mature basal body is accompanied by the cylinder elongation and appearance of pericentriolar structures, such as rootlet system, basal legs, and basal plate. Centriolegenesis proceeds in both ciliate and nonciliate (with microvilli) cells of the epithelium. It has been proposed that the cell with microvilli represent a transitional stage in differentiation of the ciliary cells.     

High herkogamy but low reciprocity characterizes isoplethic populations of Jasminum malabaricum,a species with stigma‐height dimorphism

• Studies of floral polymorphisms have focused on heterostyly, while stigma‐height dimorphism has received considerably less attention. Few studies have examined the reproductive biology of species with stigma‐height dimorphism to understand how factors influencing mate availability and pollen transfer are related to morph ratios in populations.
• Floral morphological traits, especially herkogamy and reciprocity, pollinator visitation, breeding system and spatiotemporal mate availability, are known to affect inter‐morph pollination and morph ratios in species with stigma‐height dimorphism. In this study, we investigated the presence of stigma‐height dimorphism and estimated morph ratios in four naturally occurring populations of Jasminum malabaricum. We quantified morph‐ and population‐specific differences in the abovementioned factors in these populations to understand the observed morph ratios.
• The positions of anthers and stigmas were characteristic of stigma‐height dimorphism, the first report of this polymorphism in the genus. All study populations were isoplethic, implying equal fitness of both morphs. Herkogamy was higher in the short‐styled morph, while reciprocity was higher between the long‐styled stigma and short‐styled anthers. Long‐ and short‐tongued pollinators were common floral visitors, and we observed no differences between morphs in spatiotemporal mate availability or pollinator visitation. Neither morph exhibited self‐ or heteromorphic incompatibility.
• The short‐styled stigma had lower reciprocity but likely receives sufficient inter‐morph pollen from long‐tongued pollinators, and also by avoiding self‐pollination due to higher herkogamy. These results highlight the importance of sufficient effective pollinators and floral morphological features, particularly herkogamy, in maintaining isoplethy in species with stigma‐height dimorphism.

     

Analysis of amino acids in nectar from Silene colorata Poiret (Caryophyllaceae)

Nectar samples were collected from Silene colorata Poiret (Caryophyllaceae), in three different populations from south-western Spain: Zahara de la Sierra (Cádiz), Bornos (Cádiz) and Bormujos (Seville). Samples were analysed for amino acids by reverse-phase high-performance liquid chromatography with precolumn phenylisotiocyanate (PITC) derivatization. The method has the advantage of being highly sensitive, capable of detecting nanogram (ng) quantities of amino acids. Eighteen amino acids were identified and quantified. The mean number of amino acids in a nectar sample was 14 (SD = 2.8). Six amino acids (threonine, alanine, arginine, proline, tyrosine and methionine) were detected in all samples, accounting for 83% of the total amino acids content; proline and arginine were the most abundant amino acids, accounting for 40% and 20% of the total amino acids, respectively. The mean amounts of amino acids in nectar samples per population were 824, 782 and 356 µ m in Zahara de la Sierra, Bornos and Bormujos, respectively. Environmental variations such as temperature and sunlight are factors influencing the metabolic processes of nectar production. Our results may contradict the theory that the chemical constituents of floral nectar vary according to the kinds of pollinators.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 155 , 49–56.     

Stimulation of interstitial cell growth after selective destruction of foetal Leydig cells in the testis of postnatal rats

Summary Five-day-old male rats received a single treatment of ethane dimethanesulphonate (EDS), and the response of the testis on days 6–10 and 21 was examined by light microscopy and morphometry, supplemented by measurement of peripheral testosterone levels. One day after treatment, foetal Leydig cells degenerated, showing fragmentation, condensation and nuclear pyknosis. Macrophages phagocytosed the foetal Leydig cells resulting in their disappearance by day 7. Destruction of foetal Leydig cells was followed by an arrest of testicular growth in comparison to testes of intact age-matched control rats. In testes of EDS-treated rats, gonocytes and spermatogonia also degenerated, forming pyknotic bodies within the seminiferous cords. In contrast, interstitial fibroblasts and mesenchymal cells showed proliferative activity, which on days 4 and 5 after treatment resulted in peritubular hyperplasia surrounding each seminiferous cord. Thereafter, on day 21 after EDS administration, the previously depressed serum testosterone levels became markedly elevated coincident with the development of many immature-type Leydig cells, of which the total volume per testis was similar to that of Leydig cells in control testes, despite a four- to five-fold difference in testicular volumes. The results indicate that, although EDS destroys the foetal Leydig cells and impairs spermatogenesis, the interstitial tissue exhibits increased cell growth. The latter probably occurs in response to altered gonadotrophic stimulation and/or disturbances in the interaction between the seminiferous cords and the interstitial tissue.     

Acoustic and Morphological Differentiation in the Frog Allobates femoralis: Relationships with the Upper Madeira River and Other Potential Geological Barriers

We studied patterns of call acoustics and external morphological differentiation in populations of the dart-poison frog Allobates femoralis occurring in forested areas along a 250-km stretch of the upper Madeira River, Brazil. Multivariate analyses of variance using principal components representing shared acoustic and morphological parameters distinguished three groups in relation to call structure and external morphology: (1) populations belonging to a two-note call morphotype; (2) populations with four-note calls inhabiting the left riverbank; and (3) populations with four-note calls inhabiting the right riverbank. Our results report a case of Amazonian anuran diversity hidden by current taxonomy and provide evidence for the upper Madeira River being a boundary between distinct populations of A. femoralis , and suggest a new taxonomic interpretation for these groups. Samples that did not fit into the general differentiation pattern and the existence of a well-defined contact zone between two morphotypes on the left riverbank indicate that mechanisms complementary to river-barrier hypotheses are necessary to explain the phenotypic differentiation between populations. Our study shows that at least one anuran species shows congruence between population differentiation and separation by a large Amazonian river, as documented for birds and mammals. Conservation efforts should not consider the taxon now known as A. femoralis as a homogeneous entity. There is much within-taxon variability, which can be probably explained partly by the existence of cryptic species, partly by geological barriers and part of which currently has no obvious explanation.     

Immunological detection of a cysteine protease in the skin and other tissues

Summary Monospecific antibody directed to cysteine protease of 2-day-old rat epidermis recently characterized as being different from the proteases previously reported was produced in rabbits. By immunofluorescence microscopy and immunoperoxidase staining with an avidin-biotin-peroxidase method the protease was found to be present in the epidermis of rodents of different ages as well as that of humans, but not in the dermis. The staining in germinative cells was more intense than in cells in the superficial layers. It appeared as irregular patches in the nuclei and stained more diffusely in the cytoplasm where small granular components, strongly stained, were identified. The staining patterns in granular cells showed accumulation of the antigen in a granular form. The morphology and distribution of granules resembled those of keratohyalin-like granules in the nucleus and dense homogenous deposits in the cytoplasm. In cornified cells the reaction product was localized by the plasma membrane where concentration of the dense homogenous deposits occurred, suggesting that the cysteine protease is one component of the unique and characteristic structure of differentiated keratinocytes. In addition, the cysteine protease antigen having the same molecular weight as the epidermal enzyme was detected in liver, kidney and lung indicating a wider tissue distribution of the protease. The significance of the protease in regulation of cellular functions remains to be investigated.     

Influence of retinoids and EGF on growth of embryonic mouse palatal epithelia in culture

Summary Retinoids and growth factors seem to be important for normal mammalian reproduction and development. High levels of retinoic acid are teratogenic and induce cleft palate in the mouse. Little is known concerning the mechanisms through which retinoids induce cleft palate. Palatal epithelia from CD-1 embryonic mice on Day 12 of gestation were isolated from the mesenchyme and cultured in serum-free media, with all-trans retinoic acid or 13-cis retinoic acid, with or without epidermal growth factor (EGF). The epithelia attached and grew, and the cells differentiated over a 72-h culture period. Binding of [¹²⁵I]EGF was observed in all cultures in a pattern that correlated with thymidine (TdR) uptake by the epithelia. EGF enhanced growth and [³H]TdR incorporation of the oral cells, but nasal cells generally did not proliferate. In this culture system, both retinoids suppressed [³H]TdR incorporation in a concentration-dependent manner for epithelia cultured with or without EGF. Medial cells are important to normal palatogenesis as they play a role in fusion of opposing shelves and subsequently many of these cells undergo programmed cell death. Death of medial cells in vitro is prevented by EGF and by the retinoids, either with or without EGF. This response occurs in the absence of a mesenchymal interaction, suggesting that the medial cell response to EGF and retinoids is not mediated by or dependent on the mesenchymal tissues. The survival of medial cells may be responsible for the failure of opposing shelves to fuse.