Turgor pressure is critically important for all organisms with the cell wall. In fungi, turgor is involved in the apical growth of hyphae, affects cell size, provides tension to the plasma membrane, creates the necessary rigidity for hyphae to penetrate the substrate, and has many other functions. However, there is increasing evidence that turgor pressure is not always the sole or main factor influencing some of these processes. This review characterizes the curtain model, previously proposed to describe the regulation of plasma membrane tension in the hyphae of basidiomycetes. The current understanding of the four main components of the model is outlined: the driving actin cytoskeleton, the elastic cell wall, tight adhesion of the plasma membrane to the cell wall, and macroinvaginations of the plasma membrane. All four elements, as a single model, complement or replace some physiological functions of turgor and allow us to understand how a non-apical fungal cell maintains its physiological functionality under changing environmental conditions. Further experimental confirmation of this model is fundamentally important for mycology and applied sciences.

Under certain conditions, fungi are capable to fast (down to seconds) change the size of their cells. For example, it is known that the cells of many species of yeasts under hyperosmosis instantly and reversibly shrink entirely, without plasmolysis, with a decrease in volume of up to 40% or more. There is limited data that filamentous fungi can also instantly change the diameter of their unspecialized hyphae. This property is fundamental and is a powerful tool for survival and preservation of the physiological functions of the fungus in sharply changing external conditions, but requires detailed study. In this work, which is a large-scale (more than 50,000 cells measured) light microscopic study, the ability of three unrelated basidiomycete species to fast change the diameter of their hyphae under the influence of various factors was analyzed. It was found, for example, that all three fungi respond similarly to moderate hyperosmotic shock and shrink by an average of 12-14% in diameter, but inhibitors of actin assembly can cause both expansion and shrinkage of hyphae, or have no effect on a fungus. Combining the current results with those previously obtained in the study of macroinvagination systems of the fungal plasma membrane, different approaches for basidiomycetes to respond to sudden stress changes were proposed. The previously presented curtain model has also been advanced. The model describes a general mechanism for the rapid regulation of hyphal sizes, which involves the elastic properties of the cell wall, adhesion of the plasma membrane to it, regulation of turgor, a system of actin cables that regulate the tension of the plasma membrane and cell size, as well as a complex system of macroinvaginations of the plasma membrane, ensuring constant integrity and functionality of the latter.### Competing Interest StatementThe authors have declared no competing interest.

Abstract Members of Ceratobasidiaceae family (more commonly known by its anamorphic name “rhizoctonias”) possess a variety of nutritional modes: plant pathogens, saprotrophs, endophytes and symbionts of orchid mycorrhiza. Links between nutritional modes and taxonomy of these fungi as well as their specificity towards plant host is still ambiguous. The scope of the present study was to explore biodiversity of ceratobasidioid mycobionts of sciophytic terrestrial orchid Goodyera repens, search for evolutionary stable clades within mycobionts of Goodyerinae subtribe uniform by plant host or geographic region and to establish possible connection between ceratobasidioid nutritional modes and morphological characteristics. We consider G. repens a generalist associated with a wide range of distantly related mycobionts. Two unidentified Ceratobasidium species and Thanatephorus ochraceus are reported from G. repens roots for the first time. Ceratobasidiaceae tend to form stable clades specific to either temperate or tropical region. Morphological characteristics of pathogenic and mycorrhizal rhizoctonia isolates tend to form a variety of transitional forms to correlate with nutritional mode.

The plasma membrane of filamentous fungi forms large-sized invaginations, which are either tubes or parietal vesicles. Vesicular macroinvaginations at the ultrastructural level correspond to classical lomasomes. There is an assumption that vesicular macroinvaginations/lomasomes may be involved in macrovesicular endocytosis. The original aim of this study was to test for the presence of macroendocytosis in xylotrophic basidiomycetes using time-lapse and Z-stacks fluorescent microscopic technologies. However, the results were unexpected since most of the membrane structures labeled by the endocytic tracer (FM4-64 analog) are various types of plasma membrane macroinvaginations and not any endomembranes. All of these macroinvaginations have a tubular or lamellar genesis. Moreover, under specific conditions of a microscopic preparation, the diameter of the tubes forming the macroinvaginations increases with the time of the sample observation. In addition, the morphology and successive formation of the macroinvaginations mimic the endocytic pathway; these invaginations can easily be mistaken for endocytic vesicles, endosomes, and vacuole-lysosomes. The paper analyzes the various macroinvagination types, suggests their biological functions, and discusses some features of fungal endocytosis. This study is a next step toward understanding complex fungal physiology and is a presentation of a new intracellular tubular system in wood-decaying fungi.

Like most eukaryotic organisms, fungi use endocytosis for nutrition, signal transduction, turnover of plasma membrane molecules, etc. It is generally accepted that in filamentous fungi, as in yeast, invaginations of the plasma membrane of a small size (up to about 100 nm) are formed in the early stages of endocytosis. These invaginations are surrounded by a rigid actin scaffold -an actin patch, and give rise to small primary endocytic vesicles after scission from the plasma membrane. However, in classical mycological studies, complex large-volume invaginations of the plasma membrane - lomasomes -were described in filamentous fungi. In our time, in a number of filamentous basidiomycetes when tracking endocytosis using styryl fluorescent labels, large invaginations of the plasma membrane have been found, presumably forming endocytic macrovesicles after scission. In this paper, for comparison, large-sized types of endocytosis in animal cells are briefly described. Information about tubular endocytic invaginations in fungi is presented. Three types of large invaginations of the plasma membrane, detected at the TEM level in basidiomycetes, are characterized. The main question this paper addresses is whether or not filamentous fungi do have an analogue of animal macropinocytosis -macrovesicular endocytosis. There are some indications that the answer to this question is yes, but further research is needed. The presence of macrovesicular endocytosis may change the well-established beliefs about the cellular organization of filamentous fungi and the physiology of their nutrition. (c) 2021 British Mycological Society. Published by Elsevier Ltd. All rights reserved.

The development of mycological gerontology requires effective methods for assessing the biological age of fungal cells. This assessment is based on the analysis of a complex of aging and oxidative stress markers. One of the most powerful such markers is the protein carbonylation. In this study, the already known method of dry immune dot blotting is adapted for mycological studies of the content of protein carbonyl groups. After testing the method on a number of filamentous fungi species, some features of the accumulation of carbonylated proteins in mycelium were established. Among these features: (i) a weak effect of exogenous oxidative stress on the accumulation of carbonyls in a number of fungi, (ii) reversibility of the carbonyl accumulation, (iii) possibility of arbitrary regulation of carbonyl content by fungus itself and (iv) the influence of hormesis. In addition, two polar strategies for the accumulation of carbonyl modification were revealed, named Id-strategy (Indifferent) and Cn-strategy (Concern). Thus, even the analysis of one marker allows making some preliminary general assumptions and conclusions. For example, the idea that fungi can freely regulate their biological age is confirmed. This feature makes fungi very flexible in terms of responding to environmental influences and promising objects for gerontology.

BACKGROUND INFORMATION:Two main systems regulate plasma membrane tension (PMT) and provide a close connection between the protoplast and the cell wall in fungi: turgor pressure and the actin cytoskeleton. These systems work together with the plasma membrane focal adhesion to the cell wall and their contribution to fungal cell organization and physiology has been partially studied. However, it remains controversial in model filamentous ascomycetes and oomycetes and even less investigated in filamentous basidiomycetes. Early endocytosis can be used to research the mechanisms regulating PMT since the dynamics of early endocytosis is largely dependent on this tension.RESULTS:This study examined the effects of actin polymerization inhibitors and hyperosmotic shock on early endocytosis and cell morphology in two filamentous basidiomycetes. The main obtained results are: (i) the depolymerisation of F-actin leads to the fast formation of endocytic pits while inhibiting of their scission from the plasma membrane and (ii) the moderate hyperosmotic shock does not affect the dynamics of early endocytosis. These and our other results have allowed suggesting a curtain model for the regulation of PMT in basidiomycetes.CONCLUSIONS AND SIGNIFICANCE:According to the proposed curtain model, the PMT in many non-apical cells of hyphae is more often regulated not by turgor pressure but by a system of actin driver cables that are associated with the proteins of the focal adhesion sites. The change in PMT occurs similar to the movement of a curtain along the curtain rod using the curtain drivers. This model addresses the fundamental properties of the fungal structure and physiology. It requires confirmation including the currently technically unavailable high-quality labelling of the actin cytoskeleton of the basidiomycetes.

Nitrogen in sufficient quantities is strictly necessary for all living organisms. In this study, the ability of some xylotrophic basidiomycetes to grow extremely long on a solid growth medium full of carbon nutrition but lacking a nitrogen source in its composition was discovered. The nitrogen oligotrophy of wood-decaying fungi is associated with their adaptation to live in a wood substrate, which is also deficient in nitrogen content. This nitrogen-depleted cultural growth is called "pseudo-foraging" and can be used as a simplified model of wood-decaying growth. Four main nitrogen-obtaining and -conserving strategies (nitrogen concentration, using alternative sources of nitrogen, economy of growth, and nutritional recycling), which are attributed to wood-colonizing xylotrophs in the literature, were revised studying the pseudo-foraging model. Based on the results, some aspects of the behavior of xylotrophs deep in undecomposed wood were predicted. For example, one of the results is that for pseudo-foraging xylotrophs, the main way to obtain nitrogen is its concentration in their mycelium from the nutrient medium in which nitrogen is contained in the impurities of the components of the medium. The result suggests that in bulk solid wood, the nitrogen concentration strategy also dominates the strategy of using diazotrophic and other alternative nitrogen. In addition, three individual unprecedented mechanisms, which supposedly help the xylotrophic fungi to colonize wood in nature (generation of fine mycelium, macrovesicular endocytosis, formation and conversion of super-elongated mitochondria), were investigated in the laboratory.

Alkaliphilic fungi are fundamentally different from alkalitolerant ones in terms of mechanisms of adaptation. They accumulate trehalose in cytosol and phosphatidic acids (PA) in the membrane lipids, whereas alkalitolerants contain these compounds in low amounts. But it is unclear how the composition of osmolytes and lipids changes during cytodifferentiation. In this article the composition of lipids and soluble cytosol carbohydrates in the mycelium and fruit bodies of the alkaliphilic fungus Sodiomyces alkalinus was studied. In the mycelium, mannitol and trehalose dominated, while in fruit bodies only trehalose was predominant. Phosphatidylcholines (PC), PA and sterols were major membrane lipids of the mycelium, while PC and sterols were predominant in fruit bodies. The degree of fatty acids unsaturation of the main mycelium phospholipids (PC and PA) increased with age, while that of PC did not change regardless of the developmental stage. In young mycelium, storage lipids were represented mainly by free fatty acids, and in mature mycelium and fruit bodies—by triacylglycerols. Fruit bodies contained three times less membrane lipids and twice as many storage lipids as mycelium. Trehalose was the main cytosol carbohydrate in the mycelium and fruit bodies, which confirms its key value for alkaliphily.

Exploring life cycles of fungi is insightful for understanding their basic biology and can highlight their ecology. Here, we dissected the sexual and asexual life cycles of the obligate alkalophilic ascomycete Sodiomyces alkalinus that thrives at extremely high pH of soda lakes. S. alkalinus develops acremonium-type asexual sporulation, commonly found in ascomycetous fungi. However, the sexual stage was unusual, featuring very early lysis of asci which release young ascospores inside a fruit body long before its maturation. In a young fruit body, a slimy matrix which originates from the combined epiplasm of asci and united cytoplasm of the pseudoparenchymal cells, surrounds pooled maturing ascospores. Upon maturity, the ascospores are forcibly released through a crack in the fruit body, presumably due to an increased turgor pressure. These features of the sexual stage development resemble the ones found in unrelated marine fungi, indicating convergent evolution of the trait. We hypothesise these developmental features of S. alkalinus to be adaptive in the conditions of periodically inundated rims of soda lakes where the fungus thrives.

Alkaliphily, the ability of an organism to thrive optimally at high ambient pH, has been well-documented in several lineages: archaea, bacteria and fungi. The molecular mechanics of such adaptation has been extensively addressed in alkaliphilic bacteria and alkalitolerant fungi. In this study, we consider an additional property that may have enabled fungi to prosper at alkaline pH: altered contents of membrane lipids and cytoprotectant molecules. In the alkaliphilic Sodiomyces tronii, we showed that at its optimal growth pH 9.2, the fungus accumulates abundant cytosolic trehalose (4–10% dry weight) and phosphatidic acids in the membrane lipids, properties not normally observed in neutrophilic species. At a very high pH 10.2, the major carbohydrate, glucose, was rapidly substituted by mannitol and arabitol. Conversely, lowering the pH to 5.4–7.0 had major implications both on the content of carbohydrates and membrane lipids. It was shown that trehalose dominated at pH 5.4. Fractions of sphingolipids and sterols of plasma membranes rapidly elevated possibly indicating the formation of membrane structures called rafts. Overall, our results reveals complex dynamics of the contents of membrane lipids and cytoplasmic sugars in alkaliphilic S. tronii, suggesting their adaptive functionality against pH stress.

Endocytosis is a complex process of absorption from the environment (and subsequent distribution within the cell) of soluble substances, macromolecules, microparticles, etc. by means of vesicles developed by cytoplasmic membrane. Endocytosis in animal and human cells is actively and successfully studied. Thus, classification of this process in the animals (based only on the peculiarities of primary vesicle formation) includes up to ten different endocytosis pathways. Modern knowledge about endocytosis in mycelial fungi is not so extensive; therefore, its study in this group of organisms is a topical and promising direction in fundamental and applied mycology. In the present work, we studied the effect of six different inhibitors (acting both on the assembly of actin/tubulin cytoskeleton and on the formation of different types of endocytosis) on the dynamics of endocytosis in phytopathogenic heterobasidial fungus Rhizoctonia solani. The estimation of the effect of inhibitors was conducted by means of microscopic analysis of the absorption of the fluorescent marker of endocytosis AM4-64 by mycelial cells. As a result of the conducted study, four types of the inhibitor effect on the R. solani endocytosis were detected: from the complete absence of the effect to severe suppression of different stages of fungal endocytosis. It was found that four of six inhibitors used for the suppression of endocytosis in the animals and human have a suppressive effect on endocytosis of R. solani. This indicates the conservative nature of some endocytosis mechanisms in the studied fungus and probably in mycelial fungi in general. Different hypotheses concerning principles of the effect of studied inhibitors on endocytosis activity of fungi were suggested.

The fungal strain INA 01108 producing antibiotic substances with broad spectrum of antibacterial activity was isolated from the natural environment. By the morphological characteristics and DNA analysis it was shown to belong to Ascomycetes of Sordariomycetes. In submerged culture the strain produced at least four antibiotics. The major component of them was identified as eremophilane-type sesquiterpene eremoxylarin A. Eremoxylarin A is effective in vitro against grampositive bacteria, including methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin group glycopeptide antibiotics resistant Leuconostoc mesenteroides VKPM B-4177. The efficacy and toxicity of eremoxylarin A was determined on a murine staphylococcal sepsis model. The dose of 6.25 mg/kg provided 100% recovery and survival of the animals, while the dose of 3.12 mg/kg was close to the ED50. The chemical structure of eremoxylarin A allows to modify the antibiotic and such studies may be relevant to design a less toxic derivative without loss of the valuable antimicrobial properties.