Erika Kothe

Genome sequence of the model mushroom Schizophyllum commune

Much remains to be learned about the biology of mushroom-forming fungi, which are an important source of food, secondary metabolites and industrial enzymes. The wood-degrading fungus Schizophyllum commune is both a genetically tractable model for studying mushroom development and a likely source of enzymes capable of efficient degradation of lignocellulosic biomass. Comparative analyses of its 38.5-megabase genome, which encodes 13,210 predicted genes, reveal the speciess unique wood-degrading machinery. One-third of the 471 genes predicted to encode transcription factors are differentially expressed during sexual development of S. commune. Whereas inactivation of one of these, fst4, prevented mushroom formation, inactivation of another, fst3, resulted in more, albeit smaller, mushrooms than in the wild-type fungus. Antisense transcripts may also have a role in the formation of fruiting bodies. Better insight into the mechanisms underlying mushroom formation should affect commercial production of mushrooms and their industrial use for producing enzymes and pharmaceuticals.

Siderophores mediate reduced and increased uptake of cadmium by Streptomyces tendae F4 and sunflower (Helianthus annuus), respectively

Aims:  As a toxic metal, cadmium (Cd) affects microbial and plant metabolic processes, thereby potentially reducing the efficiency of microbe or plant‐mediated remediation of Cd‐polluted soil. The role of siderophores produced by Streptomyces tendae F4 in the uptake of Cd by bacteria and plant was investigated to gain insight into the influence of siderophores on Cd availability to micro‐organisms and plants.

Involvement of siderophores in the reduction of metal-induced inhibition of auxin synthesis in Streptomyces spp

Unlike synthetic metal chelators, microbe-assisted phytoremediation provides plants with natural metal-solubilizing chelators which do not constitute a potential source of environmental pollution. Concurrently with microbial chelators, plant growth promotion can be enhanced through bacterially-produced phytohormones. In this work, the simultaneous production of siderophores and auxins by Streptomyces was studied to gain insight for future application in plant growth and phytoremediation in a metal-contaminated soil. Standard auxin and siderophore detection assays indicated that all of the investigated Streptomyces strains can produce these metabolites simultaneously. However, Al(3+), Cd(2+), Cu(2+), Fe(3+) and Ni(2+), or a combination of Fe(3+) and Cd(2+), and Fe(3+) and Ni(2+) affected auxin production negatively, as revealed by spectrophotometry and gas chromatography-mass spectrometry. This effect was more dramatic in a siderophore-deficient mutant. In contrast, except for Fe, all the metals stimulated siderophore production. Mass spectrometry showed that siderophore and auxin-containing supernatants from a representative Streptomyces species contain three different hydroxamate siderophores, revealing the individual binding responses of these siderophores to Cd(2+) and Ni(2+), and thus, showing their auxin-stimulating effects. We conclude that siderophores promote auxin synthesis in the presence of Al(3+), Cd(2+), Cu(2+) and Ni(2+) by chelating these metals. Chelation makes the metals less able to inhibit the synthesis of auxins, and potentially increases the plant growth-promoting effects of auxins, which in turn enhances the phytoremediation potential of plants.

A Consistent Phylogenetic Backbone for the Fungi

The kingdom of fungi provides model organisms for biotechnology, cell biology, genetics, and life sciences in general. Only when their phylogenetic relationships are stably resolved, can individual results from fungal research be integrated into a holistic picture of biology. However, and despite recent progress, many deep relationships within the fungi remain unclear. Here, we present the first phylogenomic study of an entire eukaryotic kingdom that uses a consistency criterion to strengthen phylogenetic conclusions. We reason that branches (splits) recovered with independent data and different tree reconstruction methods are likely to reflect true evolutionary relationships. Two complementary phylogenomic data sets based on 99 fungal genomes and 109 fungal expressed sequence tag (EST) sets analyzed with four different tree reconstruction methods shed light from different angles on the fungal tree of life. Eleven additional data sets address specifically the phylogenetic position of Blastocladiomycota, Ustilaginomycotina, and Dothideomycetes, respectively. The combined evidence from the resulting trees supports the deep-level stability of the fungal groups toward a comprehensive natural system of the fungi. In addition, our analysis reveals methodologically interesting aspects. Enrichment for EST encoded data—a common practice in phylogenomic analyses—introduces a strong bias toward slowly evolving and functionally correlated genes. Consequently, the generalization of phylogenomic data sets as collections of randomly selected genes cannot be taken for granted. A thorough characterization of the data to assess possible influences on the tree reconstruction should therefore become a standard in phylogenomic analyses.

Basidiomycete mating type genes and pheromone signaling.

ABSTRACT The genome sequences of the basidiomycete Agaricomycetes species Coprinopsis cinerea, Laccaria bicolor, Schizophyllum commune, Phanerochaete chrysosporium, and Postia placenta, as well as of Cryptococcus neoformans and Ustilago maydis, are now publicly available. Out of these fungi, C. cinerea, S. commune, and U. maydis, together with the budding yeast Saccharomyces cerevisiae, have been investigated for years genetically and molecularly for signaling in sexual reproduction. The comparison of the structure and organization of mating type genes in fungal genomes reveals an amazing conservation of genes regulating the sexual reproduction throughout the fungal kingdom. In agaricomycetes, two mating type loci, A, coding for homeodomain type transcription factors, and B, encoding a pheromone/receptor system, regulate the four typical mating interactions of tetrapolar species. Evidence for both A and B mating type genes can also be identified in basidiomycetes with bipolar systems, where only two mating interactions are seen. In some of these fungi, the B locus has lost its self/nonself discrimination ability and thus its specificity while retaining the other regulatory functions in development. In silico analyses now also permit the identification of putative components of the pheromone-dependent signaling pathways. Induction of these signaling cascades leads to development of dikaryotic mycelia, fruiting body formation, and meiotic spore production. In pheromone-dependent signaling, the role of heterotrimeric G proteins, components of a mitogen-activated protein kinase (MAPK) cascade, and cyclic AMP-dependent pathways can now be defined. Additionally, the pheromone-dependent signaling through monomeric, small GTPases potentially involved in creating the polarized cytoskeleton for reciprocal nuclear exchange and migration during mating is predicted.

Interaction of endophytic microbes with legumes.

Large numbers of bacterial and fungal endophytes have been reported from different plant tissues: roots, nodules, leaves, flowers and sprouts of legumes, with numbers ranging from few to more than 150. Endophytes can accelerate seedling emergence, promote plant establishment under adverse conditions and enhance plant growth. Endophytic microbes promote plant growth by helping plants in acquiring nutrients, e.g. via nitrogen fixation, phosphate solubilization or iron chelation, by preventing pathogen infections via antifungal or antibacterial agents, by outcompeting pathogens for nutrients by siderophore production, or by establishing the plants systemic resistance. Further growth promotion is affected by producing phytohormones such as auxin or cytokinin, or by producing the enzyme 1‐aminocyclopropane‐1‐carboxylate (ACC) deaminase, which lowers plant ethylene levels. For establishment of endophytes in different tissues, endophytic microbes must be compatible with the host plants and able to colonize the tissues of the host plants without being recognized as pathogens. A particular bacterium or fungus may affect plant growth and development using one or more of these mechanisms, and they may use different mechanisms at various times. The population density of endophytes is highly variable, depending mainly on the microbial species and host genotypes, developmental stage and environmental conditions. Genotypic and cultivar specific endophytes have also been reported. The quantum benefit derived by plants from an endophyte and vice versa is still not clear. It seems that the endophytic genus or species best adapted for living inside a plant is naturally selected. Here, we concentrate on soil or rhizosphere‐derived endophytes recruited out of a large pool of soil or rhizospheric microbes. Some endophytes are more aggressive colonizers and displace others, but seeming lack of strict specificity has been observed. However, the processes of host‐microbe signaling and colonization and the mechanisms leading to mutual benefits are less‐well characterized. It is still not clear which population of microorganisms (endophytes or rhizospheric) promotes plant growth and the way the interactions among endophytes influence plant productivity. Though attempts to know the molecular ecology and interactions are underway, a high amount of progress is required to fully understand the mechanism of establishment, the way interactions take place in planta, between different microbes and plants and exlusive benefits by endophytes and plants. (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

Evidence for high affinity nickel transporter genes in heavy metal resistant Streptomyces spec.

We have isolated 25 new strains of streptomycetes from soil samples of a polluted site at the former uranium mine, Wismut, in eastern Thuringia, Germany. The strains grew on medium containing 1 mM NiCl2 and thus were resistant to the heavy metal ion. Seven of the strains were further characterized. All of these strains were resistant to heavy metals in various degrees with up to 10 mM resistance against NiCl2 supplied with the liquid minimal growth medium. The high level of resistance prompted us to look for high affinity nickel transporter genes thought to provide a means to eliminate the excess nickel ions form the cells. Degenerate oligonucleotide primers derived from sequences of P‐type ATP‐ase transporter genes of Gram negative bacteria identified a fragment which shows deduced amino acid sequence similarities to known high affinity nickel transporters. Investigation of two genes obtained from the isolates Streptomyces spec. E8 and F4 showed high sequence divergence. This was unex‐pected since a transmissible plasmid had been thought to convey heavy metal resistance.

Mating-type genes for basidiomycete strain improvement in mushroom farming

Abstract. Mushroom production is dependent on the quality of the spawn used to inoculate the cultures. In order to produce high-quality spawn, breeding programs for strains resistant to certain diseases and able to form high-quality fruit bodies under standard growth conditions are necessary. The investigation of the molecular basis for mating provides access to the use of mating-type genes in order to facilitate breeding. For research purposes, two mushroom-forming homobasidiomycetes have been used due to their easy cultivation and sexual propagation on defined minimal media: Schizophyllum commune and Coprinus cinereus. The mating-type genes control formation of the dikaryon from two haploid strains. Only the dikaryon is fertile and able to form mushrooms under the right environmental conditions. These genes are now used in mating-type-assisted breeding programs for economically important mushrooms, especially the white button mushroom, Agaricus bisporus, and the oyster mushroom, Pleurotus ostreatus, aiming at high-yield and high-quality standard mushroom production. Most mushroom species posses two mating-type loci that control their breeding. The genes encoded in the A loci lead to the formation of transcription factors that belong to the class of homeodomain proteins. Active transcription factors are formed by heterodimerization of two proteins of different allelic specificities. In nature, this is only the case if two cells of different mating type have fused to combine the different proteins in one cytoplasm. While fusion in homobasidiomycetes is found irrespectively of mating type, exchange of nuclei between mating mycelia is dependent on the products of the B mating-type loci. The B genes form a pheromone and receptor system that enables the fungi to initiate nuclear migration. The molecular details of the two genetic systems controlling breeding in basidiomycetes are presented in this review.

Identification of a Hydrophobin Gene That is Developmentally Regulated in the Ectomycorrhizal Fungus Tricholoma terreum

ABSTRACT The symbiosis between ectomycorrhizal fungi and trees is an essential part of forest ecology and depends entirely on the communication between the two partners for establishing and maintaining the relationship. The identification and characterization of differentially expressed genes is a step to identifying such signals and to understanding the regulation of this process. We determined the role of hydrophobins produced by Tricholoma terreum in mycorrhiza formation and hyphal development. A hydrophobin was purified from culture supernatant, and the corresponding gene was identified. The gene is expressed in aerial mycelium and in mycorrhiza. By using a heterologous antiserum directed against a hydrophobin found in the aerial mycelium of Schizophyllum commune, we detected a hydrophobin in the symbiosis between T. terreum and its native pine host Pinus sylvestris. The hydrophobin was found in aerial mycelium of the hyphal mantle and also in the Hartig net hyphae, which form the interface between both partners. Interestingly, this was not the case in the interaction of T. terreum with a host of low compatibility, the spruce Picea abies. The differential expression with respect to host was verified at the transcriptional level by competitive PCR. The differential protein accumulation pattern with respect to host compatibility seen by immunofluorescence staining can thus be attributed at least in part to transcriptional control of the hyd1 gene.

Environmental and ecological interactions

Microorganisms never live alone. It thus comes as no surprise that a specific environment structures microbial communities. The specific reactions of microbes to their environment and their ecological niche, in turn, will change this environment. Within this, Special Issue articles are assembled covering the entire range of microbial interactions with their environment including rather specialized ecological interactions. The changes that microorganisms can induce are by no means small. In one of the article featured in this issue, the potential of carbonate formation is addressed (Vahabi et al., this issue). Here, an application for the sealing of fissures in concrete using inoculation strategies is discussed. This perfectly shows that the results of microbially induced reactions can have a huge effect on the surrounding environment, even if this is something as hard as a stone (or concrete) building. Other interactions are prone to change diversity. Ants in tropical forests are shown to harbour actinobacteria (Hernandez Reyes and Cafaro, this issue), and by exerting pressure on one specific clade can change community composition. In agriculture, changes in land-use are known to lead to changes in microbial communities and diversity. The application of straw is featured here (Lu et al., this issue), as well as the population degrading humic substances (Park and Kim, this issue). These soil microbiomes effect soil fertility and plant growth. A more direct interactions of microbes with plants is seen in symbiotic relationships of nodule forming bacteria with plant roots (Chen et al., this issue), and soil dwelling diazotrophswill interact as well with plants growing in the soil (Ra et al., this issue). Endophytes in roots and nodules will modulate the effect of the rhizobial symbiosis (Saini et al., this issue) and depending on environmental conditions and plant species, these root associations will vary. And plant growth promoting bacteria or rhizobacteria will exert an effect with their associated plants (Mehta et al., this issue). However, interactions of microorganisms with plants will include pathogenic interactions as well. A new pathovar leading to banana blight is shown to stress this point (Huang et al., this issue). On the other hand, microbial interactions can be used by man for plant protection. Biocontrol organisms against fungal infections are described within this Special Issues for three different cropping systems (Nguyen et al., Solanki et al., Sunar et al., all this issue). Finally, the need for new antibiotics drives research to assess the microbiomes of niches not previously exploited, like microorganisms from a soft coral featured here (ElAhwany et al., this issue). There, the environment clearly leads to a community which exerts special features.Other, extreme,environmentshavebeenstudied to yield radiation-resistant bacteria with a study on the Taklimakan Desert (Yu et al., this issue). And even within a single species, specific changes associated with environmental changes can be described, as seen with an article on fatty acid profiles within Vibrio (Xu et al., this issue). Taken together, this Special Issue gives a good example on bacterial and microorganism diversity that clearly is dependent on the different niches and ecological associations. The interactions of microorganisms with their environment, with other microbes or plants or animals in their specific niche allowed for the development of mechanisms dealing with stress, interacting in mutually symbiotic and pathogenic relationships, and even to change the environment. These features of the microbial world are driving terrestrial as well as marine environmental development.