Search

The paired antennal lobes were long considered the sole primary processing centers of the olfactory pathway in holometabolous insects receiving input from the olfactory sensory neurons of the antennae and mouthparts. In hemimetabolous insects, however, olfactory cues of the antennae and palps are processed separately. For the holometabolous red flour beetle Tribolium castaneum, we could show that primary processing of the palpal and antennal olfactory input also occurs separately and at distinct neuronal centers. While the antennal olfactory sensory neurons project into the antennal lobes, those of the palps project into the paired glomerular lobes and the unpaired gnathal olfactory center. Here we provide an extended analysis of the palpal olfactory pathway by combining scanning electron micrographs with confocal imaging of immunohistochemical staining and reporter expression identifying chemosensory and odorant receptor-expressing neurons in the palpal sensilla. In addition, we extended the anatomical characterization of the gnathal olfactory center by 3D reconstructions and investigated the distribution of several neuromediators. The similarities in the neuromediator repertoire between antennal lobes, glomerular lobes, and gnathal olfactory center underline the role of the latter two as additional primary olfactory processing centers.

Plant pathogenic fungi colonizing living plant tissue secrete a cocktail of effector proteins to suppress plant immunity and reprogramme host cells. Although many of these effectors function inside host cells, delivery systems used by pathogenic bacteria to translocate effectors into host cells have not been detected in fungi. Here, we show that five unrelated effectors and two membrane proteins from Ustilago maydis, a biotrophic fungus causing smut disease in corn, form a stable protein complex. All seven genes appear co-regulated and are only expressed during colonization. Single mutants arrest in the epidermal layer, fail to suppress host defence responses and fail to induce non-host resistance, two reactions that likely depend on translocated effectors. The complex is anchored in the fungal membrane, protrudes into host cells and likely contacts channel-forming plant plasma membrane proteins. Constitutive expression of all seven complex members resulted in a surface-exposed form in cultured U. maydis cells. As orthologues of the complex-forming proteins are conserved in smut fungi, the complex may become an interesting fungicide target., This study reports that five effectors and two transmembrane proteins from the plant pathogenic fungus Ustilago maydis form a stable cell surface-exposed protein complex required for virulence.

Morphological, anatomical and ultrastructural characteristics are important for taxonomical and phylogenetic studies of fungi. For scanning electron microscopy (SEM), usually only dry voucher specimens are available. For dried plant material, Aerosol® OT (AOT) has been shown to be a suitable rehydration agent for SEM preparation. For swelling and stabilization of fungal cells, however, this simple method does not yield satisfactory results. Here, we show that a combination of AOT with ultrasonic bath and rehydration in a vacuum desiccator is a good method to distend fungal cells like basidiospores and pleuro- and cheilocystidia for SEM analysis. Tissues of several species of Agaricomycetes with diverse morphological structures were exposed to the treatment. Diverse concentrations of AOT as well as treatments in an ultrasonic bath and a vacuum desiccator were tested to optimize the surface reconstruction and to reduce preparation artefacts. The evaluated rehydration method is a cheap, quick and nontoxic method to prepare dried specimens of fungal cells for SEM analysis.

Morphological, anatomical and ultrastructural characteristics are important for taxonomical and phylogenetic studies of fungi. For scanning electron microscopy, usually only dry voucher specimens are available. For dried plant material, Aerosol OT has been shown to be a suitable rehydration agent for SEM preparation. For swelling and stabilization of fungal cells, however, this simple method does not yield satisfactory results. Here, we show that a combination of Aerosol OT with ultrasonic bath and rehydration in a vacuum desiccator is a good method to distend fungal cells like basidiospores, pleuro- and cheilocystidia for SEM analysis. Tissues of several species of Agaricomycetes with diverse morphological structures were exposed to the treatment. Diverse concentrations of Aerosol OT as well as treatments in an ultrasonic bath and a vacuum desiccator were tested to optimize the surface reconstruction and to reduce preparation artefacts. The evaluated rehydration method is a cheap, quick and nontoxic method to prepare dried specimens of fungal cells for SEM analysis.

Guanine nucleotide exchange factors (GEF) of the BRAG subfamily activate small Arf GTPases, which are pivotal regulators of intracellular membrane traffic and actin dynamics. Consequently, BRAG proteins have been implicated to regulate the surface levels of adhesive and signaling receptors. However, not much is known about the mechanism leading to the regulation of these surface proteins. In this study we found that the Drosophila BRAG GEF Schizo interacts physically with the Abl-interactor (Abi). schizo mutants display severe defects in myoblast fusion during syncytial muscle formation and show increased amounts of the cell adhesion protein N-cadherin. We demonstrate that the schizo myoblast fusion phenotype can be rescued by the expression of the Schizo GEF (Sec7) and membrane-binding (pleckstrin homology) domain. Furthermore, the expression of the Sec7-PH domain in a wild-type background decreases the amounts of N-cadherin and impairs myoblast fusion. These findings support the notion that the Sec7-PH domain serves as a constitutive-active form of Schizo. Using a yeast-two hybrid assay, we show that the SH3 domain of Abi interacts with the N-terminal region of Schizo. This region is also able to bind to the cytodomain of the cell adhesion molecule N-cadherin. To shed light on the function of Schizo and Abi in N-cadherin removal, we employed epistasis experiments in different developmental contexts of Drosophila. These studies point towards a new model for the regulation of Schizo. We propose that the binding of Abi to the N-terminal part of Schizo antagonizes Schizo function to inhibit N-cadherin removal.

Plastic pollution has become a serious issue on Earth. Although efficient industrial recycling processes exist, a significant fraction of plastic waste still ends up in nature, where it can endure for centuries. Slow mechanical and chemical decay lead to the formation of micro- and nanoplastics, which are washed from land into rivers and finally end up in the oceans. As such particles cannot be efficiently removed from the environment, biological degradation mechanisms are highly desirable. Several enzymes have been described that are capable of degrading certain plastic materials such as polyethylene terephthalate (PET). Such enzymes have a huge potential for future biotechnology applications. However, they require model systems that can be efficiently adapted to very specific conditions. Here, we present detailed instructions, how to convert the model diatom Phaeodactylum into a solar-fueled microbial cell factory for PETase expression, resulting in a whole cell catalyst for PET degradation at moderate temperatures under saltwater conditions.

Plastic pollution has become a serious issue on Earth. Although efficient industrial recycling processes exist, a significant fraction of plastic waste still ends up in nature, where it can endure for centuries. Slow mechanical and chemical decay lead to the formation of micro- and nanoplastics, which are washed from land into rivers and finally end up in the oceans. As such particles cannot be efficiently removed from the environment, biological degradation mechanisms are highly desirable. Several enzymes have been described that are capable of degrading certain plastic materials such as polyethylene terephthalate (PET). Such enzymes have a huge potential for future biotechnology applications. However, they require model systems that can be efficiently adapted to very specific conditions. Here, we present detailed instructions, how to convert the model diatom Phaeodactylum into a solar-fueled microbial cell factory for PETase expression, resulting in a whole cell catalyst for PET degradation at moderate temperatures under saltwater conditions.

Guanine nucleotide exchange factors (GEF) of the BRAG subfamily activate small Arf GTPases, which are pivotal regulators of intracellular membrane traffic and actin dynamics. Here, we demonstrate a novel interaction between the Abl-interactor (Abi) and the BRAG family member Schizo. We mapped the SH3 domain of Abi to interact with the N-terminal region of Schizo. This region is additionally involved in the binding of the cytodomain of the cell adhesion molecule N-cadherin. Inschizoloss of function mutants, we detected increased amounts of N-cadherin. In contrast, the expression of the GEF (Sec7) and the membrane-binding (pleckstrin homology) domains decreased amounts of N-cadherin, indicating a crucial role of the Sec7-PH module in regulating N-cadherin levels. Unlike other Sec7 GEFs, where the catalytic Sec7 domain is autoinhibited, the Sec7 and PH domain of BRAG2 are constitutively accessible, raising the question how GEF activity is controlled in a spatial and temporal manner. Our genetic analyzes demonstrate that the nature of the Abi Schizo interaction is to antagonize Schizo function and to restore wild-type amounts of N-cadherin.

The biological degradation of plastics is a promising method to counter the increasing pollution of our planet with artificial polymers and to develop eco-friendly recycling strategies. Polyethylene terephthalate (PET) is a thermoplast industrially produced from fossil feedstocks since the 1940s, nowadays prevalently used in bottle packaging and textiles. Although established industrial processes for PET recycling exist, large amounts of PET still end up in the environment-a significant portion thereof in the world's oceans. In 2016, Ideonella sakaiensis, a bacterium possessing the ability to degrade PET and use the degradation products as a sole carbon source for growth, was isolated. I. sakaiensis expresses a key enzyme responsible for the breakdown of PET into monomers: PETase. This hydrolase might possess huge potential for the development of biological PET degradation and recycling processes as well as bioremediation approaches of environmental plastic waste.Using the photosynthetic microalga Phaeodactylum tricornutum as a chassis we generated a microbial cell factory capable of producing and secreting an engineered version of PETase into the surrounding culture medium. Initial degradation experiments using culture supernatant at 30 °C showed that PETase possessed activity against PET and the copolymer polyethylene terephthalate glycol (PETG) with an approximately 80-fold higher turnover of low crystallinity PETG compared to bottle PET. Moreover, we show that diatom produced PETase was active against industrially shredded PET in a saltwater-based environment even at mesophilic temperatures (21 °C). The products resulting from the degradation of the PET substrate were mainly terephthalic acid (TPA) and mono(2-hydroxyethyl) terephthalic acid (MHET) estimated to be formed in the micromolar range under the selected reaction conditions.We provide a promising and eco-friendly solution for biological decomposition of PET waste in a saltwater-based environment by using a eukaryotic microalga instead of a bacterium as a model system. Our results show that via synthetic biology the diatom P. tricornutum indeed could be converted into a valuable chassis for biological PET degradation. Overall, this proof of principle study demonstrates the potential of the diatom system for future biotechnological applications in biological PET degradation especially for bioremediation approaches of PET polluted seawater.

Additional file 1: Figure S1. Secretion analysis of PETase-GFP in P. tricornutum cultures. Figure S2. Coomassie-staining and mass spectrometry analysis of secreted PETase-FLAG in P. tricornutum cultures. Figure S3. Western Blot of a PNGase F treated protein sample (18 µl) of the total precipitated medium fraction. Figure S4. Expression and secretion efficiency analysis of AP_SP-PETaseR280A-FLAG using Western Blot. Figure S5. Scanning electron microscopic analysis of PET bottle film degradation by AP_SP-PETase-FLAG clone 1 (AP_1) secreted from P. tricornutum on a f/2 agar plate for 5 weeks. Figure S6. Scanning electron microscopic image of a P. tricornutum clone AP_SP-PETase-FLAG_1 (AP_1) cell imprint on PET bottle film incubated on a f/2 agar plate for 5 weeks. Figure S7. Scanning electron microscopic analysis of amorphous PETG film degradation by PETase-FLAG tag secreted from P. tricornutum. Figure S8. Scanning electron microscopy and UHPLC analysis of amorphous PETG film treated with 1 ml supernatant of a 500 ml culture of a P. tricornutum clone expressing AP_SP-PETase-FLAG (clone 2). Figure S9. Scanning electron microscopy and UHPLC analysis of PET (bottle) film treated with 1 ml supernatant of a 500 ml culture of a P. tricornutum clone expressing AP_SP-PETase-FLAG (clone 2). Figure S10. UHPLC with 1 ml supernatant of a 500 ml culture of a P. tricornutum clone expressing AP_SP-PETase-FLAG_2 and standard measurements. Figure S11. PET degradation experiment (UHPLC) using shredded PET as a substrate and clone AP_SP-PETase-FLAG_1. Figure S12. Predicted N-glycosylation pattern for AP_SP-PETase-FLAG by NetNGlyc 1.0.

A big challenge of the 21st century is to cope with the huge amounts of plastic waste on Earth. Especially the oceans are heavily polluted with plastics. To counteract this issue, biological (enzymatic) plastic decomposition is increasingly gaining attention. Recently it was shown that polyethylene terephthalate (PET) can be degraded in a saltwater-based environment using bacterial PETase produced by a marine diatom. At moderate temperatures, plastic biodegradation is slow and requires sensitive methods for detection, at least at initial stages. However, conventional methods for verifying the plastic degradation are either complex, expensive, time-consuming or they interfere with the degradation process. Here, we adapt lensless digital holographic microscopy (LDHM) as a new application for efficiently monitoring enzymatic degradation of a PET glycol copolymer (PETG). LDHM is a cost-effective, compact and sensitive optical method. We demonstrate enzymatic PETG degradation over a time course of 43 days employing numerical analysis of LDHM images.

The fungal genus Strobilurus belongs to Physalacriaceae and contains approximately 11 species worldwide. Species of this genus grow and reproduce on cones of various conifers, seed pods or fruits of Magnolia and Liquidambar, and branches and wood of conifers. Previous studies focused mainly on samples from Europe and North America. And no genus-specific phylogenetic analysis has been carried out to date. The monophyly, degree of species diversity and substrate specificity, and overall distribution patterns are addressed here using morphological and molecular evidence. The authors collected samples of Strobilurus from much of its known distribution ranges and carried out morphological observations and multilocus phylogenetic analyses using five molecular markers. The results show that Strobilurus is a monophyletic group but may exclude one species, S. ohshimae. A total of 13 species was identified, with two, S. orientalis and S. pachycystidiatus, described as new from China. Several species were shown to be specific to certain substrates, whereas a few less so. Biogeographic analyses indicated that historical exchanges of species between East Asia, Europe, and North America, later vicariance events, and substrate specificity have contributed jointly to diversification of Strobilurus.

Purple Laccaria are ectomycorrhizal basidiomycetes associated with temperate forests all over the Northern Hemisphere in at least two taxa: Laccaria amethysteo-occidentalis in North America, and L. amethystina complex in Eurasia, as shown by Vincenot et al. (2012). Here, we combine a further study of the genetic structure of L. amethystina populations from Europe to southwestern China and Japan, using neutral Single Sequence Repeat (SSR; microsatellite) markers; and a systematic description of two novel Asian species, namely Laccaria moshuijun and Laccaria japonica, based on ecological, morphological, and molecular criteria (rDNA sequences). Population genetics provides evidence of the ancient isolation of three regional groups, with strong signal for speciation, and suggests a centre of origin of modern populations closest to present-day Chinese populations. Phylogenetic analyses confirm speciation at the molecular level, reflected in morphological features: L. moshuijun samples (from Yunnan, China) display strongly variable cheilocystidia, while L. japonica samples (from Japan) present distinctive globose to subglobose spores and clavate cheilocystidia. This study of a species complex primarily described with an extremely wide ecological and geographical range sheds new light on the biodiversity and biogeography of ectomycorrhizal fungi.

The present paper analyses the community structure of ectomycorrhiza (ECM) and arbuscular mycorrhiza (AM) fungi associated with seven different poplar clone types growing in a patch system on soil from four different former land use types, originating from spruce forest, poplar stand, grassland and cornfield. We determined the extent to which ECM and AM play a role on the studied factors (genotype, former land use type and host growth). The diversity of ECM and AM fungal communities was estimated by morphological and molecular analyses of the 18S and ITS of the rDNA genes. Fifteen ECM fungal taxa and four AM groups were distinguished in the roots of the poplars grown for 18 months on soil originating from the respective land use types. The poplar clones showed significantly different rates of shoot length and AM colonization, especially concerning the occurrence of Glomus intraradices and Scutellospora sp. Populus deltoides had significantly higher Scutellospora sp. abundance. Although ECM abundance and diversity was high, no significant differences between the different land use types was found. However, some ECM fungi like Paxillus involutus, Laccaria proxima and Laccaria tortilis showed significant preferences for specific land use types. Our findings suggest that both factors, former land use type and poplar genotype, are important determinants of mycorrhizal colonization of the host plants.

Summary Ash dieback, caused by the pathogen Hymenoscyphus pseudoalbidus, is an emerging lethal disease of Fraxinus excelsior in large parts of Europe. To develop a method for the early detection of H. pseudoalbidus, we designed primers for 46 microsatellites (simple sequence repeats, SSRs) of the pathogen. Seven pairs of primers (SSR38, SSR58, SSR114, SSR198, SSR206, SSR211 and SSR212) were found to bind only to the genome of H. pseudoalbidus, but not to the genome of H. albidus or to 52 different fungal endophytes isolated from F. excelsior and F. angustifolia. Using these seven primer pairs, H. pseudoalbidus was identified in fruiting bodies and different types of ash tissues including dead leaves, dead petioles and discoloured or non-discoloured wood. Along one twig, H. pseudoalbidus was detected at different levels of intensity, which depended on the distance from symptomatic tissue. The detection limit was 0.9–1.8 pg of genomic DNA per PCR. Of 50 analysed commercially available seedlings, six were infected with H. pseudoalbidus. Two SSR loci (SSR198 and SSR211) showed fragment length polymorphism. Our results showed that the new primers not only provide an easy and inexpensive means of detecting H. pseudoalbidus in ash tissues, but can also provide information on the genetic heterogeneity of the species.

Phylogenetic mid-point rooted tree of the GRs based on protein sequences. Outer rings represent the expression in body, mouthparts (T. castaneum: palps, mandible, labrum, and labium; D. melanogaster: palp and proboscis; An. gambiae: maxillary palp) and antenna as a percentage compared to the highest expressed gene according to the scale in the left upper corner. Note that the methods used to obtain the different expression data (RNAseq and microarray) are not directly comparable. This figure can, thus, only give an impression of the tissue-specific abundance of the transcripts. The scale bars within the trees represent 1 amino acid substitution per site. Potential sugar and fructose receptors are labeled and highlighted in yellow and in grey, and CO2 receptors are highlighted in orange. (PDF 1733 kb)

IR gene tissue expression and chromosomal localization of IR and SNMP genes. a Venn diagram showing the number of IRs expressed (RPKMâ â Ľâ 0.5) in the different body parts: antennae, legs, mouthparts (as a piece of the head capsule anterior of the antennae), heads (the whole head capsule including mouthparts but excluding the antennae), and bodies (excluding head and legs). b Based on Georgia GA-2 strain genome assembly 3 [81], only chromosomal linkage groups containing an IR or SNMP are depicted. Gene clusters are indicated by a number referring to the chromosome and a letter conveys the relative position on the chromosome. The number of genes within this cluster is indicated in square brackets. (PDF 66 kb)

Sequences of primers and template plasmid used to generate pSLfa1180[2.5kbOrcoUp_GAL4delta]. (PDF 111 kb)

Primary and secondary antibodies and dyes used with additional information such as source and specificity. n/a not available. (PDF 118 kb)

Tomato (Solanum lycopersicum L.) roots from four different crop sites in Colombia were surface sterilized and 51 fungal isolates were obtained and conserved for further analysis. Based on microscopical observations and growth characteristics, 20 fungal isolates corresponded to genus Fusarium, six presented asexual conidia different from Fusarium, eight were sterile mycelia, seven of which had dark septate hyphae and 17 did not continue to grow on plates after being recovered from conservation. Growth on different media, detailed morphological characterization and ITS region sequencing of the six sporulating and eight sterile isolates revealed that they belonged to different orders of Ascomycota and that the sterile dark septate endophytes did not correspond to the well known Phialocephala group. Interactions of nine isolates with tomato plantlets were assessed in vitro. No effect on shoot development was revealed, but three isolates caused brown spots in roots. Colonization patterns as analyzed by confocal microscopy differed among the isolates and ranged from epidermal to cortical penetration. Altogether 11 new isolates from root endophytic fungi were obtained, seven of which showed features of dark septate endophytes. Four known morphotypes were represented by five isolates, while six isolates belonged to five morphotypes of putative new unknown species.

Microsatellite markers were established by an improved combined simple sequence repeat (CSSR) approach for the ectomycorrhizal basidiomycete Laccaria amethystina. Six markers delivered codominant polymorphic results, with up to six different alleles. They were tested with DNA originating from sporocarps and ectomycorrhizal root tips from silver fir (Abies alba), collected in the northern Black Forest. Sporocarps and ectoymcorrhizae exhibited similar allelic profiles, without any distorting influence of the host tree species. Results were compared to allelic profiles delivered by three published Asian markers. Allelic frequencies of the new markers showed a higher resolution of individuals within the population community, but partially a lower grade of heterozygosity. Possible reasons are discussed, and further questions addressed.

Here we present the first checklist of the UNESCO World Natural Heritage Kellerwald-Edersee National Park in the federal state of Hesse in Germany. After 10 years of investigation, 1107 species have been recorded. Thus far, efforts have led to the detection of 44 species of interest according to the criteria of the International Union for the Conservation of Nature and Natural Resources (IUCN), 10 species with nature value based on a European scale, and 16 nature-value species on a German scale. Sixty-four threatened species are listed on the German red list of fungi, and 147 species are listed on the regional red list of the federal state of Hesse. As part of the UNESCO World Natural Heritage Ancient Beech Forests of Germany and Primeval Beech Forests of the Carpathians, the Kellerwald-Edersee National Park represents the typical European natural beech forest on poor soil with old tree stands and relict primeval forest fragments. The presence of nature-value species in the beech (Fagus sylvatica)-dominant Kellerwald-Edersee and Hainich national parks shows the development of their forest ecosystems to near naturalness comparable to the primeval forests of the Poloniny National Park in Slovakia.

Metamorphosis is a fundamental developmental process and has been intensively studied for various neuron types of Drosophila melanogaster. However, detailed accounts of the fate of identified peptidergic neurons are rare. We have performed a detailed study of the larval morphology and pupal remodelling of identified peptidergic neurons, the CAPA-expressing Va neurons of D. melanogaster. In the larva, Va neurons innervate abdominal median and transverse nerves that are typically associated with perisympathetic organs (PSOs), major neurohaemal release sites in insects. Since median and transverse nerves are lacking in the adult, Va neurites have to undergo substantial remodelling during metamorphosis. We have examined the hitherto uncharacterised gross morphology of the thoracic PSOs and the abdominal median and transverse nerves by scanning electron microscopy and found that the complete reduction of these structures during metamorphosis starts around pupal stage P7 and is completed at P9. Concomitantly, neurite pruning of the Va neurons begins at P6 and is preceded by the high expression of the ecdysone receptor (EcR) subtype B1 in late L3 larvae and the first pupal stages. New neuritic outgrowth mainly occurs from P7-P9 and coincides with the expression of EcR-A, indicating that the remodelling of the Va neurons is under ecdysteroid control. Immunogold-labelling has located the CAPA peptides to large translucent vesicles, which are released from the transverse nerves, as suggested by fusion profiles. Hence, the transverse nerves may serve a neurohaemal function in D. melanogaster.

Within the basidiomycetes, the vast majority of known mycorrhizal species are homobasidiomycetes. It was therefore surprising when molecular and ultrastructural studies revealed a broad diversity of mycorrhizal associations involving members of the heterobasidiomycetous Sebacinaceae, fungi which, due to their inconspicuous basidiomes, have been often overlooked. To investigate the phylogenetic position of the Sebacinaceae within the basidiomycetes and to infer phylogenetic relationships within the Sebacinaceae, we made molecular phylogenetic analyses based on nuclear rDNA. We present a well-resolved phylogeny of the main lineages of basidiomycetes which suggests that the Sebacinaceae is the most basal group with known mycorrhizal members. Since more basal taxa of basidiomycetes consist of predominantly mycoparasitic and phytoparasitic fungi, it seems possible that a mycorrhizal life strategy, which was transformed into a saprotrophic strategy several times convergently, is an apomorphic character for the Hymenomycetidae. Mycorrhizal taxa of Sebacinaceae, including mycobionts of ectomycorrhizas, orchid mycorrhizas, ericoid mycorrhizas, and jungermannioid mycorrhizas, are distributed over two subgroups. One group contains species with macroscopically visible basidiomes, whereas members of the other group probably lack basidiomes. Sebacina appears to be polyphyletic; current species concepts in Sebacinaceae are questionable. Sebacina vermifera sensu Warcup & Talbot consists of a broad complex of species possibly including mycobionts of jungermannioid and ericoid mycorrhizas. This wide spectrum of mycorrhizal types in one fungal family is unique. Extrapolating from the known rDNA sequences in Sebacinaceae, it is evident that there is a cosm of mycorrhizal biodiversity yet to be discovered in this group. Taxonomically, we recognise the Sebacinaceae as constituting a new order, the Sebacinales.

Species of Tricholoma sect. Pardinicutis (Singer) Bon are relatively easily recognizable even in the field, and the type species of section, T. pardinum (Pers.) Quél., was reported from the eastern Himalaya and adjacent areas. However, such reports were largely based on superficially similar morphology. In this study, we have generated DNA sequences of samples from southwestern China, and found that there are molecular discrepancies between the Chinese collections and European ones. Further detailed morphological analyses indicated the two independent new species occur in southwestern China, one in subtropical coniferous forests mixed with fagaceous plants between 2400 and 2800 m altitude, the other in subalpine dark coniferous forests between 3300 and 4100 m altitude. Consequently, two new species, namely, T. highlandense and T. sinopardinum, are described and illustrated.

In this paper descriptions of three new Laccaria species from Southwest China (Yunnan) are reported. Macromorphological, micromorphological, and molecular data have been studied to describe the new species and delineate them within the genus Laccaria. The first species Laccaria fulvogrisea is characterized by a grey to brownish coloured fruiting body and large echinulate spores. Laccaria aurantia is characterized by the deeply orange colour and globose balloon-like spores with a fine echinulate ornament. The third species Laccaria yunnanensis has large barrel-shaped pleurocystidia and a brownish to flesh-coloured fruiting body. All three species have 4-spored basidia and were found near or within Quercus and Lithocarpus mixed broad leaved forests at an altitude of above 2,000 m.

The genus Singerocybe (Tricholomataceae, Agaricales, Basidiomycota) has been the subject of controversy since its proposal in 1988. Its taxonomic foundation, species circumscription and geographical distribution have not yet been examined with molecular sequence data. In this study phylogenetic analyses on this group of fungi were conducted based on collections from Europe, eastern Asia, southern Asia, North America and Australia, with four nuclear markers, ITS, nrLSU, tefl-alpha and rpb2. Molecular phylogenetic analyses, together with morphological observations, strongly support Singerocybe as a monophyletic group and identify the vesicles in the pileal and stipe cuticle as a synapomorphy of this genus. Seven species are recognized in the genus, including one new species and four new combinations. Clitocybe trogioides and Clitocybe trogioides var. odorifera are synonyms of Singerocybe humilis and Singerocybe alboinfundibuliformis respectively. Most of these species are geographically restricted in their distributions. Furthermore our study expands the distribution range of Singerogbe from the North Temperate Zone to Australia (Tasmania) and tropical southern Asia.

The ontogeny of the fruit bodies of Mycena stylobates was studied by light microscopy and scanning electron microscopy (SEM). The ontogeny was divided into two phases: first the primordium with all the structures of the mature fruit body was established, then the primordial stipe elongated rapidly and the exposed hymenium started producing spores immediately. The first detected stage of fruit body formation was an irregularly arranged hyphal structure within the substrate colonized. After the rupture of the surface of the substrate, the primordium established on the surface. Soon a layer of wrapping hyphae was differentiated, which covered the complete primordium. The structures of the stipe and the cap developed synchronously. The developing stipe, cap and basal disc together formed a secondary ring-like cavity, in which the development of the hymenophore took place. The lower side of the cap was covered by a layer of degenerated hyphae. The development of the hymenophore started with a number of small alveolae on the lower side of the cap, which were covered with a hymenophoral palisade. The margins of these alveolae formed the primary lamellae, which in the first stage of their development were covered by a layer of degenerated hyphae. The hymenophoral palisade spread from the developing alveolae to the lamellar edge; the edge of the primary lamellae was forked in the early stages. Secondary lamellae were formed by the down folding of ridges from the lower side of the cap. In contrast to the primary lamellae, they were covered with hymenophoral palisade from the beginning. Spore production started immediately after the elongation of the stipe. These results were compared with other known modes of ontogeny within the Agaricales and some comments on the terminology used for the description of basidiome morphogenesis are made.

A review is given about the morphological and ultrastructural characters of Piriformospora indica. The morphology of hyphal cells, mycelial, and chlamydospores are described in detail. Piriformospora is able to form different types of interactions with plant groups mosses, ferns, and gymno- and angiosperms. A survey of the tested host plants is given. The morphology of interacting hyphae and root cells depends on the plant species. Besides the mutualistic mode of life, Piriformospora indica is able to live as an endophytic fungus; under specific conditions, this species can behave as a necrotrophic fungus. All these different characters of the plant growth-promoting fungus, Piriformospora indica, indicate that this species is very valuable for basic research and a wide spectrum of applications in agriculture and horticulture.

The arbuscular mycorrhiza (AM) is characterized by specific morphological structures of the fungus and the plant and by physiological adaptations which are mostly beneficial for both partners of the symbiosis. This review describes approaches to study the molecular basis of the interaction. RNA accumulation patterns have been monitored in Pisum sativum to analyse the plant response to arbuscule development. In a direct approach, the Mtha1 gene from Medicago truncatula was cloned which is expressed in arbusculated cells of M. truncatula. The gene putatively encodes an H(+)-ATPase involved in the improved plant nutrition during mycorrhization. Finally, a tripartite system between M. truncatula, Glomus mosseae and Aphanomyces euteiches was established, in order to study bioprotection. Analysis of the transcriptome has been started to analyse the interaction between the plant, the pathogen and the AM fungus.

Piriformospora indica (Sebacinaceae, Basidiomycota) is an axenically cultivable, plant growth promoting root endophyte with a wide host range, including Populus. Rooting of Populus Esch5 explants started within 6 days after transfer to WPM medium. If such plantlets with roots were inoculated with P. indica, there was an increase in root biomass, and the number of 2nd order roots was increased significantly. A totally different observation was recorded when the explants were placed into WPM with pre-grown P. indica. The interaction led to complete blocking of root production and severely inhibited plant growth. Additionally, branched aerial roots appeared which did not penetrate the medium. On contact with the fungal colony or the medium, the ends of the aerial roots became inflated. Prolonged incubation stimulated the fungus to colonize aerial parts of the plant (stem and leaves). Mycelium not only spread on the surface of the aerial parts, but also invaded the cortical tissues inter- and intracellularly. Detached Populus leaves remained vital for 4 - 5 weeks on sterile agar media or on AspM medium with pre-grown P. indica. When the fungus was pre-grown on culture media such as WPM, containing ammonium as the main source of nitrogen, leaves in contact with the cultures turned brownish within 4 - 12 h. Thereafter, the leaves bleached, and about one day later had become whitish. Thus, cultural conditions could alter the behaviour of the fungus drastically: the outcome of the interaction between plant and fungus can be directed from mutualistic to antagonistic, characterized by fungal toxin formation and extension of the colonization to Populus shoots.

Soil microflora influence plant growth and health both beneficially and adversely. The microorganisms used as biofertilisers stimulate plant growth response by providing necessary nutrients as a result of their colonisation of the rhizosphere (Azotob acter, Azospirillum, phosphate-solubilising bacteria and cyanobacteria) or by symbiotic association (Rhizobium, mycorrhizae fungi and Frankia). They may also regulate the physiological processes in the ecosystems by involvement in the decomposition of organic matter, fixation of atmospheric nitrogen, secretion of growth-promoting substances, increase in the availability of mineral nutrients, immobilisation of these assimilable nutrients and protection of plants from pathogens (Mukerji et al. 1998). Thus, rhizosphere effects through microbial activities, modify the plants by providing plant growth substances and increasing the availability of nutrients at the root zone. Plant root anatomy and tissue articulation play a significant role in the symbiotic processes (Lynch 1995).

The larvae of the cabbage root fly induce serious damage to cultivated crops of the family Brassicaceae. We here report the biochemical characterisation of neuropeptides from the central nervous system and neurohemal organs, as well as regulatory peptides from enteroendocrine midgut cells of the cabbage maggot. By LC-MALDI-TOF/TOF and chemical labelling with 4-sulfophenyl isothiocyanate, 38 peptides could be identified, representing major insect peptide families: allatostatin A, allatostatin C, FMRFamide-like peptides, kinin, CAPA peptides, pyrokinins, sNPF, myosuppressin, corazonin, SIFamide, sulfakinins, tachykinins, NPLP1-peptides, adipokinetic hormone and CCHamide 1. We also report a new peptide (Yamide) which appears to be homolog to an amidated eclosion hormone-associated peptide in several Drosophila species. Immunocytochemical characterisation of the distribution of several classes of peptide-immunoreactive neurons and enteroendocrine cells shows a very similar but not identical peptide distribution to Drosophila. Since peptides regulate many vital physiological and behavioural processes such as moulting or feeding, our data may initiate the pharmacological testing and development of new specific peptide-based protection methods against the cabbage root fly and its larva.

A new fungus isolate was discovered in an arbuscular mycorrhizal fungal spore from a desert soil in India. It could easily be cultivated on various synthetic media, and formed pear-shaped chlamydos...

The adult musculature in D. melanogaster forms during metamorphosis. Much is known about the flight and leg musculature, but not about the muscles surrounding the male reproductive tract. The inner genitalia of males consist of the testes, which emerge from the gonads; the remaining genital organs, i.e., paragonia (or accessory glands), ejaculatory duct, sperm pump, and seminal vesicles, develop out of the genital imaginal disc. We analyzed the myoblasts forming the muscle layers of these organs. In myoblasts derived from the genital imaginal disc, the regulatory region of the transcription factor DMef2 is active. DMef2 is also needed for specification and differentiation of embryonic and adult myoblasts. We could discriminate three different muscle types: (i) multinucleated muscles that resemble vertebrate smooth muscles surround the testes, (ii) multinucleated muscles that resemble striated muscles comprises seminal vesicles and the sperm pump, and (iii) mononucleated striated musculature encloses the paragonia and ejaculatory duct. Members of the immunoglobulin superfamily involved in embryonic myogenesis, Dumbfounded (Duf) and Sticks and Stones (Sns), were also expressed in the genital imaginal disc, in the muscle sheath of the testes during muscle differentiation and in the secretory secondary cells, which are part of the binucleated epithelia enclosing the paragonia.