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.     

The role of sterols in morphogenetic processes and dimorphism in fungi

The review considers the fundamental biological problem of fungal dimorphism as an adaptive reaction to adverse impacts. Primary attention is paid to sterols, phospholipids, storage lipids, and fatty acids. The structural and biological functions of sterols are considered, as well as their role in membrane stabilization under stress and their relation to morphogenetic processes in mycelial fungi, of which many are pathogenic. Data on the biosynthesis of the main fungal sterol, ergosterol, are presented, as well as data on the inhibitors of this process and on the mutants deficient in its particular stages. Ergosterol biosynthesis is also considered in terms of its relation to the composition of the fungal cell wall, which is the cell shape-determining structure, and to the intensity of chitin synthesis, a process in which azole derivatives play a role. Data obtained by the authors are presented that show the role of changes in the composition of sterols, phospholipids, storage lipids, and unsaturated fatty acids of resting cells in the induction of yeastlike growth in mucoraceous fungi.     

The role of sterols in morphogenetic processes and dimorphism in fungi

The review considers the fundamental biological problem of fungal dimorphism as an adaptive reaction to adverse impacts. Primary attention is paid to sterols, phospholipids, storage lipids, and fatty acids. The structural and biological functions of sterols are considered, as well as their role in membrane stabilization under stress and their relation to morphogenetic processes in mycelial fungi, of which many are pathogenic. Data on the biosynthesis of the main fungal sterol, ergosterol, are presented, as well as data on the inhibitors of this process and on the mutants deficient in its particular stages. Ergosterol biosynthesis is also considered in terms of its relation to the composition of the fungal cell wall, which is the cell shape-determining structure, and to the intensity of chitin synthesis, a process in which azole derivatives play a role. Data obtained by the authors are presented that show the role of changes in the composition of sterols, phospholipids, storage lipids, and unsaturated fatty acids of resting cells in the induction of yeastlike growth in mucoraceous fungi.     

Extracellular proteases of mycelial fungi as participants of pathogenic processes

The interest in proteases secreted by mycelial fungi is due to several reasons of which one of the most important is their involvement in the initiation and development of the pathogenic process. A comparison of saprophytic and phytopathogenic mycelial fungi revealed one characteristic feature, namely, the appearance of a new trypsin-like activity in phytopathogens that is absent in saprophytes. To clear up the question of whether the degree of pathogenicity of a fungus is related to the activity of secreted trypsin-like protease, several species of Fusarium of various pathogenicity were compared. In two species, F. sporotrichioides (which causes ear fusa-riosis of rye) and F. heterosporum (the causative agent of root rot in wheat), a clear correlation between the activity and pathogenicity was revealed: the more pathogenetic F. sporotrichioides exhibited a higher extracellular trypsin-like activity than the less pathogenetic species F. heterosporum. Thus, the presence of trypsin-like activity in a saprotroph-pathogen pair may be an indicator of the pathogenicity of a fungus; in some cases, the value of this activity may indicate the degree of its pathogenicity. This suggests that trypsin-like proteases specific to phytopathogens are directly involved in the pathogenetic process, probably, through interaction with the "sentry" protein or the product of the resistance gene.     

The effect of different heat influences on composition of membrane lipids and cytosol carbohydrates in mycelial fungi

Comparison of the changes in the composition of the membrane lipids and soluble cytosol carbohydrates caused by two types of heat influence (within the tolerance zone and heat shock-level) revealed fundamental differences in the stress response of fungal cells. In three fungal species, Aspergillus niger, Pleurotus ostreatus, and Cunninghamella japonica, increased levels of trehalose and phosphatidic acids were observed under heat shock, while heat influences within the tolerance zone had no such effect. Under heat shock, the ratio of saturated fatty acids did not increase in any of the major phospholipids of all the studied species. This is in contradiction with the existing hypothesis and confirms the previously suggested the hypothesis of membrane stabilization by heat-protecting compounds.     

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.     

About different mechanisms of the biochemical adaptation of mycelial fungi to temperature stress: changes in lipids composition

A comparative study was conducted concerning the effect of temperature stress on the lipid composition of representatives of the subkingdoms Eomycota and Neomycota. Changes in the composition of lipid acyl chains (such as saturation and desaturation, isomerization, and changes in the length of fatty acid carbon chains), in the phospholipid composition, and in the contents of sterols and other neutral lipids were revealed. Hyperthermia resulted in (i) an increase in the phosphatidylcholine level; (ii) a decrease in the phosphatidylethanolamine level; (iii) a rise in the content of reserve lipids (triacylglycerols); and (iv) a decline in the free fatty acid level in the neutral lipids. An inverse pattern occurred under hypothermic conditions. The peculiarities in the patterns of the temperature adaptation-related changes in the lipid bilayer composition are considered in terms of the systematic position of the fungi.     

Choline: Its role in the growth of filamentous fungi and the regulation of mycelial morphology

Abstract Choline is an essential metabolite for the growth of filamentous fungi. It occurs most notably as a component of the major membrane phospholipid, phosphatidyl choline (lecithin), and fulfills a major role in sulphate metabolism in the form of choline- o -sulphate in many species. Choline is usually synthesised endogenously, but exogenous choline can also be taken up, either to compensate for metabolic deficiencies in choline-requiring mutants such as those of Aspergillus nidulans and Neurospora crassa , or as a normal function by species such as Fusarium graminearum which do not require added choline for growth. F. graminearum has a highly specific constitutive uptake system for this purpose. Recent studies have begun to indicate that choline also plays an important role in hyphal and mycelial morphology. Over a wide range of concentrations, choline influences mycelial morphology, apparently influences mycelial morphology, apparently by controlling branch initiation. At high concentrations of added choline, branching is inhibited but specific growth rate is unaffected, leading to the production of rapidly extending, sparsely branched mycelia. Reduction of choline concentration allows a progressive increase in branching. Additionally, in choline-requiring mutants which have a very reduced content of choline, multiple tip-formation and apical branching occurs. Just prior to cessation of growth in choline-starved cultures of A. nidulans choline-requiring mutants, hyphal morphology changes due to a brief phase of unpolarised growth to produce spherical swellings called ballons, at or near hyphal apices. The precise mechanism by which choline affects fungal morphology is not yet known, although in A. nidulans it appears to be at least partially due to the influence of membrane composition on the synthesis of the hyphal wall polymer chitin. Several hypotheses for the possible mode of action of choline in affecting fungal morphology are discussed here.     

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.     

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.     

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.     

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.     

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.     

Strategies of food substrate colonization by mycelial fungi

This article deals with a new research area related to mycology and the food industry. It is concerned with the strategies used by ascomycete mycelial fungi to utilize and colonize food items. The data of scanning and transmission electron and light microscopy revealed that fungi growing on such food items as cheese and sausage are characterized by the following colonization strategies: (i) spreading growth; (ii) burrowing growth; (iii) growth with the formation of up to 4–5 new branches on the apexes of the old hyphae; (iv) growth with the formation of long strands, and (v) growth with biofilm formation.