Edith Stabentheiner

Basidiomycete yeasts in the cortex of ascomycete macrolichens

Lichens assemble in three parts Lichen growth forms cannot be recapitulated in the laboratory by culturing the plant and fungal partners together. Spribille et al. have discovered that the classical binary view of lichens is too simple. Instead, North American beard-like lichens are constituted of not two but three symbiotic partners: an ascomycetous fungus, a photosynthetic alga, and, unexpectedly, a basidiomycetous yeast. The yeast cells form the characteristic cortex of the lichen thallus and may be important for its shape. The yeasts are ubiquitous and essential partners for most lichens and not the result of lichens being colonized or parasitized by other organisms. Science, this issue p. 488 Complete functioning lichen thalli have three partners: alga and ascomycete, plus a basidiomycete yeast. For over 140 years, lichens have been regarded as a symbiosis between a single fungus, usually an ascomycete, and a photosynthesizing partner. Other fungi have long been known to occur as occasional parasites or endophytes, but the one lichen–one fungus paradigm has seldom been questioned. Here we show that many common lichens are composed of the known ascomycete, the photosynthesizing partner, and, unexpectedly, specific basidiomycete yeasts. These yeasts are embedded in the cortex, and their abundance correlates with previously unexplained variations in phenotype. Basidiomycete lineages maintain close associations with specific lichen species over large geographical distances and have been found on six continents. The structurally important lichen cortex, long treated as a zone of differentiated ascomycete cells, appears to consistently contain two unrelated fungi.

Functional and Mutational Analysis of P19, a DNA Transfer Protein with Muramidase Activity

Protein P19 encoded by the conjugative resistance plasmid R1 has been identified as being one member of a large family of muramidases encoded by bacteriophages and by type III and type IV secretion systems. We carried out a mutational analysis to investigate the function of protein P19 and used in vivo complementation assays to test those of several P19 mutants. The results indicated that conserved residues present in the presumed catalytic center of P19 are absolutely essential for its function in conjugation of plasmid R1 and infection by the RNA phage R17. Overexpression of protein P19 in an early growth phase resulted in a massive lysis of Escherichia coli cells in liquid culture, as indicated by a rapid and distinct decrease in cell culture densities after induction. Change of the proposed catalytic glutamate at position 44 to glutamine completely abolished this effect. P19-induced cell lysis was directly shown by transmission and scanning electron microscopy. Typically, P19-overexpressing cells showed bulges protruding from the cell surfaces. Our interpretation is that these protrusions arose from a localized and spatially confined disruption of the bacterial cell wall. To our knowledge such an effect has not previously been documented for any member of the lytic transglycosylase family. From the data presented here, we conclude that protein P19 possesses the proposed localized peptidoglycan-hydrolyzing activity. This activity would be a prerequisite for efficient penetration of the cell envelope by the DNA translocation complex encoded by the conjugative plasmid.

Compartment specific response of antioxidants to drought stress in Arabidopsis

Graphical abstract

Photobiont association and genetic diversity of the optionally lichenized fungus Schizoxylon albescens.

The fungus Schizoxylon albescens occurs both as lichen and as saprobe. Lichenized colonies grow on the bark of Populus tremula; saprotrophic morphs grow on dead Populus branches. We wanted to (1) test whether lichenized and saprotrophic S. albescens are genetically distinct, (2) investigate photobiont association and diversity, (3) investigate the interactions between fungi and algae that occur during co-cultivation and (4) test whether Schizoxylon shows algal selectivity during lichenization. Fungal and algal genetic diversity were investigated for three markers. Algae from lichenized thalli were isolated in axenic cultures, and isolate sequence diversity was compared with algae amplified directly from thallus fragments. Co-culture experiments of fungi and algae were performed to study the morphological interaction patterns. Two distinct phylogenetic units are revealed in S. albescens, which are interpreted as phenotypically cryptic species. The algae are related to Coccomyxa and Pseudococcomyxa, and form two distinct sister clades separating samples isolated in cultures from those amplified directly from thallus fragments, indicating that more easily cultured strains of algae are not necessarily major components of the lichens. Schizoxylon albescens interacts with isolated algal strains, similar to fungal-Coccomyxa symbioses in nature. As the system is maintained without difficulty in culture, it can potentially be an easily controlled lichen symbiosis study system under laboratory conditions.

In-vivo Candida biofilms in scanning electron microscopy.

Candida biofilms on indwelling devices are an increasing problem in patients treated at intensive care units. The goal of this study was to examine the occurrence and frequency of these biofilms. A total of 172 catheters were collected from 105 male and 67 female patients (the age range of both patient groups was from 3 weeks to 98 years old). The catheters were incubated on blood agar plates and the resulting yeast colonies were subsequently identified. Furthermore, pieces of catheters were fixed, dried and sputter coated with gold for investigation with scanning electron microscopy (SEM). Yeasts were recovered from significantly more catheters obtained from men than from women (chi(2): n = 67; P < 0.01). In SEM, 56.4% catheters turned out to be positive for biofilm formation. Again catheters from male patients were statistically significant (chi(2): n = 40; P < 0.01) more often positive than those from women. Candida albicans (71.1%) was the most common species isolated from the catheters, followed by C. glabrata (10.3%), C. parapsilosis (8.2%) and C. tropicalis (5.2%). Based on the results of this investigation, the epidemiology of Candida biofilms on indwelling devices seems to be a promising target for future investigations.

Chrysomelidial in the opisthonotal glands of the oribatid mite, Oribotritia berlesei.

Gas chromatographic–mass spectrometric analyses of whole body extracts of Oribotritia berlesei, a large-sized soil-dwelling oribatid mite, revealed a consistent chemical pattern of ten components, probably originating from the well-developed opisthonotal glands. The three major components of the extract were the iridoid monoterpene, (3S,8S)-chrysomelidial (about 45% of the extract), the unsaturated hydrocarbon 6,9-heptadecadiene, and the diterpene β-springene (the latter two, each about 20–25% of the extract). The remaining minor components (together about 10% of the extract) included a series of hydrocarbons (tridecene, tridecane, pentadecene, pentadecane, 8-heptadecene, and heptadecane) and the tentatively identified 9,17-octadecadienal. In contrast, analysis of juveniles showed only two compounds, namely a 2:1 mixture of (3S,8S)-chrysomelidial and its epimer, epi-chrysomelidial (3S,8R-chrysomelidial). Unexpectedly, neither adult nor juvenile secretions contained the so-called astigmatid compounds, which are considered characteristic of secretions of oribatids above moderately derived Mixonomata. The chrysomelidials, as well as β-springene and octadecadienal, are newly identified compounds in the opisthonotal glands of oribatid mites and have chemotaxonomic potential for this group. This is the first instance of finding chrysomelidials outside the Coleoptera.

Structure and ultrastructure of Pinus canariensis needles.

Summary The present study using light, transmission and scanning electron microscopy contains information about the structure of the needles of Pinus canariensis , an endemic pine of the Canary Islands. As a three-needled species, they have a triangular shape in transverse section being the abaxial side hemispherical. The deeply sunken stomata and the waxes covering the epidermal cells as well as the epistomatal chamber, constitute a good adaptation to the loss of water. The mechanical tissue below the epidermis contributes to a higher drought resistance and to maintain the shape of the long needles as a skeletic tissue. The ultrastructure of mesophyll cells, transfusion tissues as well as endoderrnis and resin ducts is also described. The results of this study should be used as reference data for further investigations of physiological, biochemical and ultrastructural responses of this species to environmental variations due to pollutants and different altitudinal and exposure situations.

Bartheletia paradoxa is a living fossil on Ginkgo leaf litter with a unique septal structure in the Basidiomycota

Bartheletia paradoxa, a basidiomycete growing on fallen leaves of Ginkgo biloba, is redescribed. In autumn a rapidly developing anamorph is formed on freshly fallen leaves and subsequently a teleomorph with hemispherical pustules of thick-walled resting spores (teliospores) that germinate after a resting period of one year with stipitate, longitudinally septate, statismosporic phragmobasidia. The basidia produce several basidiospores on each sporogenous locus. Inoculation experiments and observations in the field suggest that the basidiospores infect the freshly fallen leaves of G. biloba so that the life cycle is completed. The extraordinarily rapid development has also been confirmed in cultures on agar media and in inoculation experiments. Inoculation experiments also indicate that the fungus is specific to G. biloba. The septa of the hyphae have no central pores, but multiple plasmodesma-like perforations. The basidiospores and conidia are uninucleate, but an assessment of the karyology is still pending. A molecular phylogenetic hypothesis based on nuSSU rDNA sequences suggests that the fungus belongs to the Agaricomycotina, clustering in an unresolved position at the basal branching of the group. The family Bartheletiaceae fam. nov. is proposed to accommodate Bartheletia paradoxa in the Agaricomycotina. The name B. paradoxa is validated by a Latin diagnosis and by the designation of types.

Different surface characteristics of primary and secondary needles of Pinus canariensis

Summary Surface characteristics of primary and secondary needles of Pinus canariensis were investigated using scanning electron micro-scopy and gas chromatography to study structure and composition of epicuticular wax, cuticle micromorphology and the structure of stomata. Tubular waxes could be observed on the whole needle surface of the glaucous primary needles whereas on secondary needles they were restricted to the lower surface of young needles. Recrystallization resulted in comparable wax tubes and, addition-ally, plate like structures recrystallized from primary needle wax. Isolated cuticles of primary needles were tender and showed a simple stomata complex with six subsidiary cells whereas the cuticles of secondary needles were massive and revealed 9–12 subsidiary cells. In contrast to the cuplike epistomatal chamber of the primary needles that of the secondary needles was larger and often irregularly formed. Main constituents of the cuticular wax were ω-hydroxy-n-alkanoic acids, 10-nonacosanol and n-alkanoic acids with no differences in the qualitative composition between primary and secondary needles but with some differences in the quantitative pattern. The possible role of the investigated cuticular features in adaptive strategies of the needles to avoid light and water stress is discussed.

Chemotaxonomic and micromorphological traits of Satureja montana L. and S. subspicata Vis. (Lamiaceae)

Satureja montana and S. subspicata are used as spice, pepper substitute, for preparing tea, juice, and as a medicine. Fourteen populations (seven per species) of Satureja montana L. and S. subspicata Vis. growing in Croatia were examined to determine the chemical composition of the essential oil (analyzed by GC‐FID and GC/MS), the content of macroelements (Na, K, Ca, Mg) and trace elements (B, Fe, Cu, Mn, Zn, Al, Pb, Cr, Cd, Ni, Hg, As) analyzed by ICP‐AES, antioxidant compounds (analyzed by UV/VIS spectrophotometer), and the types and distribution of trichomes (analyzed by scanning electron microscopy). The main constituents of the essential oil were carvacrol and thymol in S. montana (all populations belong to one phenol chemotype), while α‐eudesmol, β‐eudesmol, and spathulenol dominated in S. subspicata (three chemotypes could be distinguished). Both species possess considerably higher quantities of Ca and Mg, and moderate concentrations of K and Na, while Hg and As levels were below the limit of quantification. Non‐glandular trichomes, peltate trichomes, and three types of capitate trichomes were observed on leaves, stem, calyx, and corolla.