A new function of trehalose and the peculiarities of lipid formation in mycelial fungi

This work deals with lipid formation in ascomycete fungi and the effect of preservatives on them. A new biological function of trehalose was revealed, and of particular interest was the fact that the effect of this disaccharide depended on its concentration in the growth medium. In the presence of a preservative such as potassium sorbate (PS), low trehalose concentrations suppressed the growth of mycelial fungi contaminating hard cheeses and contributed to the prolongation of the preservative’s effect. A tenfold increase in trehalose concentration in the medium, conversely, resulted in a drastic increase in growth activity and removed the PS effect. Therefore, trehalose can function as an inhibitor or a stimulator of growth processes, depending on its concentration. It was established that the secondary growth of Penicillium fungi during their ontogeny is accompanied by consumption of accumulated reserve lipids. In contrast, this phenomenon does not occur in mucorous fungi, and this probably accounts for the fact that Mucorales representatives can accumulate significant triacylglyceride amounts during the idiophase.     

A model for the germ tube formation and mycelial growth form of Candida albicans

A model based on morphological and ultrastructural evidence is presented which illustrates a novel and hitherto undescribed pattern of germ tube formation and hyphal growth in early and mature colonies of Candida albicans. Accordingly, most of the cytoplasm within the parent yeast cell migrates into and forward with the extending germ tubes and leaves behind an extensively vacuolated yeast cell. Growing hyphae similarly are subtended by migrating "slugs' of protoplasm and leave behind vacuolated intercalary compartments. The vacuolated cell compartments apparently must first regenerate their protoplasmic contents before producing branches or secondary germ tubes. This model is used to explain certain unusual features of the growth kinetics of the filamentous form of this organism.     

Fungal lipid accumulation and development of mycelial structures by two arbuscular mycorrhizal fungi

We monitored the development of intraradical and extraradical mycelia of the arbuscular mycorrhizal (AM) fungi Scutellospora calospora and Glomus intraradices when colonizing Plantago lanceolata. The occurrence of arbuscules (branched hyphal structures) and vesicles (lipid storage organs) was compared with the amounts of signature fatty acids. The fatty acid 16:1omega5 was used as a signature for both AM fungal phospholipids (membrane constituents) and neutral lipids (energy storage) in roots (intraradical mycelium) and in soil (extraradical mycelium). The formation of arbuscules and the accumulation of AM fungal phospholipids in intraradical mycelium followed each other closely in both fungal species. In contrast, the neutral lipids of G. intraradices increased continuously in the intraradical mycelium, while vesicle occurrence decreased after initial rapid root colonization by the fungus. S. calospora does not form vesicles and accumulated more neutral lipids in extraradical than in intraradical mycelium, while the opposite pattern was found for G. intraradices. G. intraradices allocated more of its lipids to storage than did S. calospora. Thus, within a species, the fatty acid 16:1omega5 is a good indicator for AM fungal development. The phospholipid fatty acid 16:1omega5 is especially suitable for indicating the frequency of arbuscules in the symbiosis. We propose that the ratio of neutral lipids to phospholipids is more important than is the presence of vesicles in determining the storage status of AM fungi.     

Growth and metabolism in mycelial fungi

A mathematical model for the growth and morphogenesis in colonies of mycelial fungi is given. The model consists of partial differential equations for accumulation of hyphae by apical growth, uptake of nutrient, and redistribution of a derived metabolite within the mycelium. Mechanisms for nutrient absorption and for metabolite translocation are discussed. An explanation for growth in the form of concentric mycelial rings is offered, based on the hypothesis that repeated metabolite buildup and depletion gives rise to different local branching rates, and thus distinct bands of hyphal densities.     

The lipids of mycelial fungi and the prospects for the development of microbial oleo-biotechnology. I. The lipids of mycelial fungi

The importance of mycelial fungi as a source of lipids for development of modern oleo biotechnology is considered. For this purpose the data on fungal lipid composition and the main enzymes of fatty acid synthesis in fungi as compared with procaryotes and higher eucaryotes as well the data on the effect of a number of physicochemical factors on the yield and composition of the lipids of the micromycetes are presented. The information on the fungi producers according to the lipid composition able to substitute the vegetable oil and be the source of linolic, linolenic and arachidonic acids is put forward.     

A fractal model for the characterization of mycelial morphology

A new technique based on a fractal model has been developed for the quantification of the macroscopic morophology of mycelia. The morphological structuring is treated as a fractal object, and the fractal dimension, determined by an ultrasonic scattering procedure developed for the purpose, serves as a quantitative morphological index. Experimental observations reported earlier and simulations of mycelial growth, carried out using a probabilistic-geometric growth model developed for the purpose, both validate the applicability of the fractal model. In experiments with three different species, the fractal dimensions of pelletous structures were found to be in the range 1.45-2.0 and those of filamentous structures were in the range 1.9-2.7, with values around 2.0 representing mixed morphologies. Fractal dimensions calculated from simulated mycelia are in rough agreement with these ranges. The fractal dimension is also found to be relatively insensitive to the biomass concentration, as seen by dilution of the original broths. The relation between morphology and filtration properties of the broths has also been studied. The fractal dimension shows a strong correlation with the index of cake compressibility and with the Kozeny constant, two filtration parameters that are known to be morphology dependent. This technique could thus be used to develop correlations between the morphology, represented by the fractal dimension, and important morphology-dependent process variables. (c) 1993 John Wiley & Sons, Inc.     

Species composition of food-spoiling mycelial fungi

We investigated the composition of the microflora that spoils foodstuffs (the surface of hard cheeses and sausages) at agribusiness factories. Mycelial fungi, mostly ascomycetes of the order Eurotiales belonging to the genus Penicillium play the main role in spoiling food. Most representatives of these fungi are mesophiles and possess the capacity for utilizing nutrient substrates in surface and submerged cultures.     

On a simplified model for pattern formation in honey bee colonies

We present a simplified version of a previously presented model (Camazine et al. (1990)) that generates the characteristic pattern of honey, pollen and brood which develops on combs in honey bee colonies. We demonstrate that the formation of a band of pollen surrounding the brood area is dependent on the assumed form of the honey and pollen removal terms, and that a significant pollen band arises as the parameter controlling the rate of pollen input passes through a bifurcation value. The persistence of the pollen band after a temporary increase in pollen input can be predicted from the model. We also determine conditions on the parameters which ensure the accumulation of honey in the periphery and demonstrate that, although there is an important qualitative difference between the simplified and complete models, an analysis of the simplified version helps us understand many biological aspects of the more complex complete model. Corresponding author     

Ectomycorrhizal fungi: exploring the mycelial frontier

Ectomycorrhizal (ECM) fungi form mutualistic symbioses with many tree species and are regarded as key organisms in nutrient and carbon cycles in forest ecosystems. Our appreciation of their roles in these processes is hampered by a lack of understanding of their soil-borne mycelial systems. These mycelia represent the vegetative thalli of ECM fungi that link carbon-yielding tree roots with soil nutrients, yet we remain largely ignorant of their distribution, dynamics and activities in forest soils. In this review we consider information derived from investigations of fruiting bodies, ECM root tips and laboratory-based microcosm studies, and conclude that these provide only limited insights into soil-borne ECM mycelial communities. Recent advances in understanding soil-borne mycelia of ECM fungi have arisen from the combined use of molecular technologies and novel field experimentation. These approaches have the potential to provide unprecedented insights into the functioning of ECM mycelia at the ecosystem level, particularly in the context of land-use changes and global climate change.     

Liposome-mediated mycelial transformation of filamentous fungi

Liposome-mediated transformation is common for cells with no cell wall, but has very limited usage in cells with walls, such as bacteria, fungi, and plants. In this study, we developed a procedure to introduce DNA into mycelium of filamentous fungi, Rhizopus nigricans LH 21 and Pleurotus ostreatus TD 300, by liposome-mediation but with no protoplast preparation. The DNA was transformed into R. nigricans via plasmid pEGFP-C1 and into P. ostreatus via 7.2 kb linear DNA. The mycelia were ground in 0.6 M mannitol without any grinding aids or glass powder for 15 min to make mycelial fragments suspension; the suspension was mixed with a mixture of the DNA and Lipofectamine 2000, and placed on ice for 30 min; 100 μL of the transformation solution was plated on potato dextrose agar (PDA) plate and cultivated at 28 °C for transformant screening. The plasmid and the linear DNA were confirmed to be integrated into the host chromosome, proving the success of transformation. The transformation efficiencies were similar to those of electroporation-mediated protoplast transformation (EMPT) of R. nigricans or PEG/CaCl₂-mediated protoplast transformation (PMT) of P. ostreatus, respectively. The results showed that our procedure was effective, fast, and simple transformation method for filamentous fungi.     

A mathematical model for the growth of mycelial pellet populations

In liquid culture, filamentous organisms often grow in the form of pellets. Growth result in an increase in radius, whereas shear forces result in release of hyphal fragments which act as centers for further pellet growth and development. A previously published model for pellet growth of filamentous microorganisms has been examined and is found to be unstable for certain parameter values. This instability has been identified as being due to inaccuracies in estimating the numbers of fragments which seed the pellet population. A revised model has been formulated, based on similar premises, but adopting a finite element approach. This considers the population of pellets to be distributed in a range of size classes. Growth results in movement to classes of increasing pellet size, while fragments enter the smallest size class, from which they grow to form further pellets. The revised model is stable and predicts changes in the distribution of pellet sizes within a population growing in liquid batch culture. It considers pellet growth and death, with fragmentation providing new centers of growth within the pellet population, and predicts the effects of shear forces on pellet growth and size distribution. Predictions of pellet size distributions are tested using previously published data on the growth of fungal pellets and further predictions are generated which are suitable for experimental testing using cultures of filamentous fungi or actinomycetes. (c) 1995 John Wiley & Sons, Inc.     

Mathematical modelling of intercellular regulation causing the formation of spatial structures in bacterial colonies

Bacterial colonies may grow forming stable spatial, particularly circular, structures. For instance, motile bacteria Proteus vulgaris or Escherichia coli grown on agar under certain conditions may form concentric rings with the centre in the inoculation point (Rüss-Münzer, 1935, Bact. Parasit Kde (Abt 1) 7, 214; Budriené, 1985, Dokl. Acad. Nauk SSR, 283, 470). A similar picture can be observed in a different situation, i.e. when a lawn of non-motile Salmonella typhimurium bacteria is cultivated on a solid agar with the locally introduced substrate (Hoppensteadt & J?ger, 1980, Lecture Notes in Biomath. 38, 68). This paper describes a mechanism of bacterial interactions through a hypothetical mediator released by the organisms. A mathematical model has been built. Its analysis has shown that the selected laws of secretion and reception of the mediator can adequately account for the formation of circular structures in the case of both motile and non-motile bacteria.     

Functional consequences of nutrient translocation in mycelial fungi

Fungi are of fundamental importance for plant and microbial nutrition with primary roles in decomposition and nutrient recycling. They also have great potential for use in areas of biotechnology such as bioremediation of organic and inorganic pollutants and biocontrol of plant pathogens. In all these contexts, environmental heterogeneity has a strong influence on growth and function. A large class of fungi overcome the difficulties encountered in such environments by the mechanism of translocation which results in the internal redistribution of nutrients within the fungal mycelium. In this paper, we use a combination of experimental techniques and mathematical modelling to examine fungal growth in general, and in particular, translocation in the common soil saprophytic fungus Rhizoctonia solani. A detailed mathematical model is presented where translocation is considered to have both diffusive and metabolically-driven components. A calibration experiment provided the necessary parameter values. Growth experiments were compared with model solutions and thus we provide strong evidence that diffusion is the dominant mechanism for translocation in homogeneous environments. In heterogeneous environments, we conclude that diffusion is still vital for exploration, i.e. the expansion of the fungal network into the surrounding area. However, we also conclude that localized resources may be utilized faster if energy is invested, i.e. when exploitation of the fungal microenvironment is enhanced by metabolically driven translocation.     

A model for pellet size distributions in submerged mycelial cultures

Fungal liquid cultures differ from those of bacteria in that clumps, called pellets are formed. Diffusional limitations constrain growth to the surface of such clumps. Previous models for pellet growth have neglected the effect of clump size distributions on growth rates. The model derived by Edelstein (1981) can be used to approach this question, and to demonstrate that fragmentation can accelerate the overall biomass growth. Experimental observations reported in this paper are in agreement with one version of this model incorporating loss of part of the pellet population due to mechanical damage or washout.     

A revised method for the observation of conidiogenous structures in fungi

Preparation of fungal material for microscopical examination of their conidiogenous structures is described below. A modification to the existing methods used coupled with a sporulation medium has proved to be simple to use and successful for the induction of sporulation and subsequent examination of conidiogenous structures in a range of xylariaceous species and in endophytic fungi.     

Diversity of facultatively anaerobic microscopic mycelial fungi in soils

The numbers of microscopic fungi isolated from soil samples after anaerobic incubation varied from tens to several hundreds of CFU per one gram of soil; a total of 30 species was found. This group is composed primarily of mitotic fungi of the ascomycete affinity belonging to the orders Hypocreales (Fusarium solani, F. oxysporum, Fusarium sp., Clonostachys grammicospora, C. rosea, Acremonium sp., Gliocladium penicilloides, Trichoderma aureoviride, T. harzianum, T. polysporum, T. viride, T. koningii, Lecanicillum lecanii, and Tolypocladium inflatum) and Eurotiales (Aspergillus terreus, A. niger, and Paecilomyces lilacimus), as well as to the phylum Zygomycota, to the order Mucorales (Actinomucor elegans, Absidia glauca, Mucor circinelloides, M. hiemalis, M. racemosus, Mucor sp., Rhizopus oryzae, Zygorrhynchus moelleri, Z. heterogamus, and Umbelopsis isabellina) and the order Mortierellales (Mortierella sp.). As much as 10–30% of the total amount of fungal mycelium remains viable for a long time (one month) under anaerobic conditions.