The Development and Behaviour of Mycelial Strands in Merulius lacrymans (Wulf.) Fr.: II. Hyphal Behaviour during Strand Formation

The system of hyphal branching by Merulius lacrymans was observedin mycelium which had grown from a wood food-base on to glassslides during incubation in sterile moist chambers. A hierarchyof branches and sub-branches arose from the region of clampconnexions, or nodes, of relatively wide main hyphae. Therewas evidence that the sequence of branches occurring at nodesin basipetal succession represented the time sequence of branchdevelopment at any one node. Later-formed branches at any nodewere smaller than earlier branches, but such earlier branchesusually became smaller towards the tip as growth continued.Mycelial strands were built up by growth and branching of thigmo-tropicallysensitive ‘tendril’ hyphae in association with thewide main hyphae. Tendril hyphae were characteristically narrow,thin-walled hyphae arising both as later-formed branches fromthe nodes of the main hyphae and as the narrowed tips of earlierbranches. Although this branching behaviour could be seen amongstaerial hyphae growing over agar media, hyphae growing in contactwith or within the agar behaved differently and did not formstrands.     

Internal mycelium of Pseudoperonospora cubensis, the causal agent of cucurbit downy mildew

The internal mycelium of Pseudoperonospora cubensis has been observed in transmission and scanning electron microscopic preparations. The internal mycelium may be inter- or intracellular. Haustoria of short swollen bundles of hyphae have been observed. Actively growing hyphae contain numerous mitochondria, nuclei, active dictyosomes, low amounts of storage materials (lipid) and microbody-like structures with a laminate inclusion. Thick walled hyphae with a diameter which is smaller than the actively growing hyphae have been observed. These thick wallcd hyphae contain large amounts of reserve material (lipid) and it is suggested that they may function as resting propagules.     

Age-dependent differential responses ofSaprolegnia hyphal tips to a helical growth-inducing factor in the agar substitute,gellan

Saprolegnia ferax produces more-or-less straight, subapically branched, hyphae when growing in liquid or agar-solidified media, with abundant aerial mycelium on the latter. In Contrast, the same medium solidified with gellan gum induced helical growth with reduced branching and almost no aerial mycelium. Helical growth induction was gellan concentration-dependent, peaking at 0.4–0.6% (w/v), when about 60% of tips were helical. Gellan-induced helices showed concentration-dependent inhibition by agarose and polyethylene glycol. Colonies on gellan-agarose, where helices were inhibited, reverted to having aerial mycelium, whereas those on gellan-polyethylene glycol did not. Branches on helical hyphae were initially linear, but converted to helical growth after about 2 h of extension. This transition was often marked by a branch, thus branch and helix competency appeared to be related. Germinating cysts took twice as long as hyphal inocula before producing helical hyphae, reinforcing the suggestion that helix competence was age-related.Achlya, but notPhytophthora, also showed gellan-induced helical growth and aerial mycelium suppression. These results showed (a) that morphogenic regulators of hyphal growth responded to gelling agents, probably high-molecular-weight polysaccharides, (b) that all growing hyphal tips were not equivalent, and (c) that hyphal tips underwent age-related changes in their response to the environment. The gellan-related differences in aerial mycelium mimic hydrophobin-based mycelium behavior and may thus indicate environmental regulation of hydrophobin production.     

Role of Fungal Mycelium in the Formation of Carbonate Concretions in Growing Media—An Investigation by SEM and Synchrotron-Based X-Ray Tomographic Microscopy

Soil fungi can facilitate calcification. Mushroom Morchella sp . mycelium induced the formation of carbonate concretions on the surface of an organic-based growing media amended with sand and ground limestone. According to SEM observation and X-ray-tomographic microscopy a dense mycelial network induced calcification. The CaCO₃ content of concretions (?: 0.3–1.5 cm) was found to be at 30%. Microsparitic calcite cemented the pores between the sand grains forming a dense clogging microstructure. Besides water uptake by the mycelium, a high evaporation rate and a decrease in pCO₂ contributed to the formation of the concretions. Fungal mycelium in the concretions is surrounded by voids indicating that at the surface of the mycelium, calcification is counteracted most probably by the release of organic acids.     

An ascomycetous endophyte isolated from Mentha piperita L.: biological features and molecular studies

A hyaline sterile fungus forming epiphyllous mycelial nets was isolated from meristem cultures of Mentha piperita. Histological studies indicated that the culture isolate is able to colonize stems and leaves with no damage to the host plant. In vitro-grown peppermint plants displayed enhanced vegetative growth when infected by the fungus, with mycelium extending from green tissues to growing rootlets. The production of very thin hyphae growing away from host meristems and the asymptomatic nature of the symbiosis were commonly observed in cultures, where the isolate never sporulated. No attribution to a precise morphospecies was therefore possible and the fungal culture was named sterile mycelium PGP-HSF. Through comparison of the 18 S rDNA sequence of the epibiont to those available in literature and in GenBank we were able to determine that the mutualist of peppermint is a member of the Pyrenomycetes, belonging to the subclass Sordariomycetidae.     

Perithecial formation inGelasinospora reticulispora V. Microscopically versus photobiologically recognizable differentiation of hyphae

Morphological changes in the early development of photobiologically induced perithecium weve microscopically observed with hyphae in apically growing mycellum ofGelasinospora reticulispora. No morphological differentiation was recognized in any hyphae during the inductive darkness and the following lag period. Semispherical protrusion was first noticed on slender hyphae 7 to 9 hr after the photoinductive treatment, and it developed into a coil-shaped mass within 3 to 6 hr thereafter. This hyphal mass grew to a young perithecium having 50 to 100 μm in diameter ca. 24 hr after the photoinduction. The density of induced pertithecium was dependent upon spot size of irradiation; the smaller the exposing area of mycelium, the higher the density. The result indicated that most slender hyphae can be responsive to photobiological induction of perithecium.     

THE SUBEPIDERMAL FUNGI OF CEREAL GRAINS II. THE NATURE, IDENTITY AND ORIGIN OF THE MYCELIUM IN WHEAT

An internal fungal mycelium between the epidermis and cross-layer cells in normal wheat grains is shown to consist of extracellular septate hyphae forming a network on the inner surface of the epidermis, whose cells are occasionally penetrated. The mycelium, which is first observed at a comparatively late stage of maturation of the grain, usually remains as a loose network, although sclerotia and plate-like structures suggestive of drought forms are occasionally present. These appearances support the view, derived from comparison of the drying rates of grain under various climatic conditions, that the spread of the subepidermal mycelium is finally restricted by desiccation beneath the epidermis during the drying out of the ripening grain.
Cultures from surface-sterilized grains of Bersée wheat have shown that the most common subepidermal fungus is Alternaria tenuis (in 64.4% of the grains). Bacteria (28.8%), Mycogone sp. (?) (7.7%), Cladosporium herbarum (5.8%), Pullularia pullulans (4.8%), Fusarium sp. (1.9%), Botrytis cinerea (1.9%) and Stemphylium botryosum (0.9%) were also isolated.
The subepidermal mycelium apparently arises either from systemic infection of the wheat plant or from fungal spores and hyphae present on the outside of the developing grains and among the dead floral parts.     

The growth of external AM fungal mycelium in sand dunes and in experimental systems

We estimated the biomass and growth of arbuscular mycorrhizal (AM) mycelium in sand dunes using signature fatty acids. Mesh bags and tubes, containing initially mycelium-free sand, were buried in the field near the roots of the dune grass Ammophila arenaria L. AM fungal mycelia were detected at a distance of about 8.5 cm from the roots after 68 days of growth by use of neutral lipid fatty acid (NLFA) 16:1ω5. The average rate of mycelium extension during September and October was estimated as 1.2 mm day⁻¹. The lipid and fatty acid compositions of AM fungal mycelia of isolates and from sand dunes were analysed and showed all to be of a similar composition. Phospholipid fatty acids (PLFAs) can be used as indicators of microbial biomass. The mycelium of G. intraradices growing in glass beads contained 8.3 nmol PLFAs per mg dry biomass, and about 15% of the PLFAs in G. intraradices, G. claroideum and AM fungal mycelium extracted from sand dunes, consisted of the signature PLFA 16:1ω5. We thus suggest a conversion factor of 1.2 nmol PLFA 16:1ω5 per mg dry biomass. Calculations using this conversion factor indicated up to 34 μg dry AM fungal biomass per g sand in the sand dunes, which was less than one tenth of that found in an experimental system with Glomus spp. growing with cucumber as plant associate in agricultural soil. The PLFA results from different systems indicated that the biomass of the AM fungi constitutes a considerable part of the total soil microbial biomass. Calculations based on ATP of AM fungi in an experimental growth system indicated that the biomass of the AM fungi constituted approximately 30% of the total microbial biomass. This revised version was published online in June 2006 with corrections to the Cover Date.     

Mycophenolic Acid Production by Penicillium brevicompactum on Solid Media

When grown on Czapek-Dox agar, Penicillium brevicompactum produced mycophenolic acid after a vegetative mycelium had been formed and as aerial hyphae were developing. Nutrients were still plenteous in the agar when the synthesis began. If aerial hyphal development was prevented by placing a dialysis membrane over the growing fungus, no mycophenolic acid was produced. When the dialysis membrane was peeled back and, as a consequence, production of aerial hyphae began, mycophenolic acid biosynthesis was observed. We concluded that mycophenolic acid was produced only by P. brevicompactum colonies that possessed an aerial mycelium.     

ENZYME ACTIVITY AND CELL DIFFERENTIATION IN NEUROSPORA

Zalokar , Marko . (Yale U., New Haven, Conn.) Enzyme activity and cell differentiation in Neurospora. Amer. Jour. Bot, 46(7): 555–559. Illus. 1959.—Morphological differences were observed in vegetative cells of Neurospora of different ages and in different parts of the mycelium. The surface layer of mycelium grown in standing cultures could be separated from the deep layer. The first contained most of the growing hyphae rich in protoplasm, while the second contained heavily vacuolated hyphae laden with fat droplets. Specific activities of several enzymes were studied in conidia, young hyphae, and the surface and deep layers of mature mycelium. Succinic dehydrogenase was low in conidia and about 10 times more active in mature mycelium. The surface layer had twice the activity of the deep layer. Aldolase increased about 3 times after the germination of conidia; it was slightly lower in the surface than in the deep layer of mycelium. Tryptophan synthetase exhibited only small differences between conidia and mycelium and was slightly lower in the surface than in the deep layer. β-galactosidase was formed in appreciable amounts only after prolonged growth and had a much higher specific activity in the deep layer. The results were discussed in connection with cell differentiation and aging.     

内生真菌司氏角担子菌B6对尖孢镰刀菌西瓜专化型菌株的拮抗作用

近年来,尽管西瓜产业不断发展壮大,但轮作土壤种植西瓜易产生枯萎病害导致世界范围内的西瓜严重减产。通过对峙培养实验和抑菌实验探讨了重阳木内生真菌司氏角担子菌(Ceratobasidum stevensii)B6菌株对西瓜枯萎病病原菌尖孢镰刀菌西瓜专化型(Fusarium oxysporum f. sp.Niveum,FON)的拮抗作用,并初步分析了其作用机制。平板拮抗试验的结果表明,内生真菌B6生长过程中不是通过产生抑菌带来抑制FON菌丝,而是利用自身的生长优势将FON完全覆盖。显微观察B6与FON菌丝的接触部位,发现FON菌丝外侧附着B6顶端菌丝形成的胞样结构,表明FON菌丝生长仅受到B6菌丝的抑制。抑菌试验结果显示,B6产生的挥发性物质可以抑制FON的生长和产孢,并使其菌丝分枝明显减少;B6的发酵液对FON的生长和产孢没有抑制作用。因此,推测B6主要通过释放某些挥发性物质产生拮抗作用而抑制FON的生长。     

Succinate dehydrogenase activity of external and internal hyphae of a vesicular-arbuscular mycorrhizal fungus,Glomus mosseae (Nicol. & Gerd.) Gerdmann and Trappe,during mycorrhizal colonization of roots of leek (Allium porrum L.), as revealed by in situ histochemical staining

The succinate dehydrogenase (SDH) activity of hyphae of the vesicular-arbuscular (VA) mycorrhizal fungus Glomus mosseae (Nicol. & Gerd.) Gerdmann and Trappe, in symbiotic association with leek (Allium porrum L.) roots, was investigated by histochemical staining in situ. Leek seedlings were transplanted to sand culture and inoculated with spores of G. mosseae placed just below the base of the stem. At intervals (14, 25, 35 and 60 days) after transplanting, the growth medium of seedlings was flooded with nitro blue tetrazolium chloride solution, thereby displacing the nutrient solution. This allowed sites of SDH activity of external and internal fungal structures of the mycorrhizas to be stained without physically disturbing the symbiotic system. After counterstaining harvested roots and mycelium with acid fuchsin, it was possible to differentiate clearly metabolically active and inactive regions of the fungus. The lengths of external hyphae and infected root both increased nearly exponentially, and were in constant proportion (1.4 m hyphae per cm of infected root) for up to 60 days. The percentage length of external hyphae with SDH activity remained almost constant (80%). In each infected length of root there was a gradation of SDH activity from inactive distal (older) hyphae to uniformly active proximal (younger) hyphae. These findings are discussed in relation to the symbiotic activity of the mycobiont.Deceased     

Microfungi of a Danish Beech forest Microbiology of a Danish beech forest II

The fungal flora in different parts of a beech forest ecosystem was investigated through a four year period as part of an IBP project. Both colony counts and direct measurements of fungal mycelium indicated that a vast majority of the fungal biomass is concentrated in the upper horizons of the soil, especially in the mull layer. The litter also contained large amounts of fungi when calculated per g dry weight, but still the litter fungi accounted for only a quite small percentage of the total fungal biomass. The fungi growing in direct contact with the living plants, i.e. in the rhizosphere and phylloplane, also accounted for only a few per cent of the total amount of fungi in the ecosystem.
On basis of direct measurements of fungal mycelium the total biomass was estimated to be about 100 g dry wt per m². However, no attempts were made to distinguish between living and dead hyphae, and a large proportion of the observed hyphae may very well be dead or inactive.
Qualitative studies revealed that the upper soil layers not only contained the largest amounts of fungal mycelium, but also by far the highest species diversity. Other parts of the ecosystem, e.g. the phylloplane, were often strongly dominated by one or a few species, whereas soil always contained a large variety of different types.     

Soil conditions and the take-all disease of wheat: VIII. Further experiments on the survival of Ophiobolus graminis in infected wheat stubble

Assimilable nitrogen in various forms prolonged the life of Ophiobolus graminis in infected wheat straw, whether added directly to the straw or to the surrounding soil. When the infected straws were buried in washed quartz sand, 0.5 g. nitrogen per 100 g. air-dry straw was the optimum dressing for longevity of Ophiobolus. Addition of sodium phosphate did not significantly increase longevity.
Nitrogen is considered to prolong the life of Ophiobolus by enabling the mycelium to form new branch hyphae, which can explore unexhausted parts of the substrate; it is suggested that aged mycelium dies from carbohydrate starvation, through exhaustion of the zones of enzymic erosion around the hyphae. This hypothesis is supported by the extended life of the fungus in infected straws that were shaken twice weekly in 3 % dextrose solution.
Ophiobolus was found to survive longer in infected straws buried in a fallow soil than in the same soil under oats, mustard or trefoil; this finding suggests the use of catch crops as competitors with Ophiobolus for soil nitrogen.     

Glutamine metabolism during aerial mycelium growth of Neurospora crassa

During vegetative growth, glutamine is accumulated in the mycelium of Neurospora crassa. This high pool of glutamine seems to be required for aerial mycelium growth. Enzymes responsible for the synthesis and catabolism of glutamine were measured before and during the partial transformation of a mycelial mat into aerial mycelium. In the transforming mycelial mat,considerable activities of the biosynthetic NADP-glutamate dehydrogenase and glutamine synthetase (predominantly β polypeptide) and also some activity of glutamate synthase were observed. In the aerial mycelium, glutamine synthetase (predominantly β polypeptide) was detected, but very low activities of NADP-glutamate dehydrogenase and glutamate mycelium could derive from glutamine. No glutaminase activity could be detected. It is suggested that glutamate is formed through the activities of the glutamine transaminase-ω -amidase pathway and another transaminase. High activities of glutamine and alanine transaminases were observed in the aerial mycelium. These results are discussed in terms of the possible role of glutamine as a nitrogen carrier from the mycelium to the growing aerial hyphae.     

EXPERIMENTS ON THE EFFECT OF CENTIMETRE-LENGTH ELECTROMAGNETIC WAVES ON WOOD-DESTROYING FUNGI

In an attempt to devise a practical method of killing wood-destroying fungi, mycelia of Coniophora puteana and Merulius lacrymans were exposed to the beamed radiation from a standard Service radar transmitter transmitting pulses of radiation of wave-length 9.1 cm. Growth of Coniophora puteana on malt agar films on glass slides was checked. Many superficial hyphae were killed. Exposures up to I hr. did not kill the mycelium of either fungus when growing on malt agar slopes or plates, or wood blocks. The only obvious effect was a check in growth whenever drying of the mycelium occurred.
It is concluded that the rays have no lethal effect and injure the fungi only through drying of the mycelium due to the increase in temperature which they bring about.     

Growth of Glomus intraradices and its effect on linseed (Linum usitatissimum L.) in hydroponic culture

This paper presents a hydroponic system for culturing and maintaining the VAM fungus Glomus intraradices in symbiosis with linseed (Linum usitatissimum L.) under greenhouse conditions in pure nutrient solution. It was possible to obtain large quantities of mycorrhizal host plant roots as well as extramatrical mycelium and chlamydospores free of impeding residues of solid substrate components. Starting from linseed donor plants inoculated in sand and transferred to the nutrient solution, new infections arose within the fast growing root system, hyphae spread out into the liquid and infected mycorrhiza-free receptor plants. Data for infection rates and plant growth parameters are presented. In comparsion to other culture systems for VAM fungi, the advantages of this hydroponic system are discussed and potential uses suggested.     

Cytoplasmic Continuity Revisited: Closure of Septa of the Filamentous Fungus Schizophyllum commune in Response to Environmental Conditions

Background

Mycelia of higher fungi consist of interconnected hyphae that are compartmentalized by septa. These septa contain large pores that allow streaming of cytoplasm and even organelles. The cytoplasm of such mycelia is therefore considered to be continuous.

Methodology/Principal Findings

Here, we show by laser dissection that septa of Schizophyllum commune can be closed depending on the environmental conditions. The most apical septum of growing hyphae was open when this basidiomycete was grown in minimal medium with glucose as a carbon source. In contrast, the second and the third septum were closed in more than 50% and 90% of the cases, respectively. Interestingly, only 24 and 37% of these septa were closed when hyphae were growing in the absence of glucose. Whether a septum was open or closed also depended on physical conditions of the environment or the presence of toxic agents. The first septum closed when hyphae were exposed to high temperature, to hypertonic conditions, or to the antibiotic nourseothricin. In the case of high temperature, septa opened again when the mycelium was placed back to the normal growth temperature.

Conclusions/Significance

Taken together, it is concluded that the septal pores of S. commune are dynamic structures that open or close depending on the environmental conditions. Our findings imply that the cytoplasm in the mycelium of a higher fungus is not continuous perse.     

A discovery of riphean heterotrophs in the Lakhanda group of Siberia

New fossil remains have been discovered from the well-known Lakhanda Microbiota (1015–1025 Ma, Uchur-Maya Region, Southeastern Siberia). The microfossils have characters observed in microscopic fungi, such as a reticulate mycelium, formed by anastomizing hyphae of non-cellular structure and of variable width. The fungal mycelium shows that hyphae grew in a certain direction, both forwards and backwards. As their length increased, the hyphae became curved and merged with neighboring hyphae to form enclosed spaces. The presence on hyphae of knoblike trapping structures and secretion of adhesive enzymes suggest a similarity with modern nematophagous fungi. Microorganisms associated with the adhesive hyphae consist of colonial green unicellular algae and thin bacterial sheaths, which may indicate mutually beneficial relationships between morphologically and biologically different partners. Microfossils of fungal origin are systematically described. They include Aimonema ramosa gen. et sp. nov. and the green unicellular alga Eoprotoderma neruenica gen. et sp. nov., forming a symbiotic association.