Hyphal Sheath Research Articles

Eight new species of lichenized Basidiomycota in the genera Acantholichen, Cyphellostereum and Dictyonema s.str. (Agaricales, Hygrophoraceae) from northern South America

A study of Acantholichen P.M. Jørgensen, Cyphellostereum D.A. Reid and Dictyonema s.str. C. Agardh ex Kunth from the Venezuelan Andes (Sierra Nevada de Mérida) and the Guayana Highlands (Alto Orinoco) revealed the presence of eight species new to science, viz., Acantholichen dendroideus V. Marcano, Cyphellostereum mucuyense V. Marcano, Dictyonema andinum V. Marcano, D. duidense V. Marcano, D. laurae V. Marcano, D. subinvolutum V. Marcano, D. subsericeum V. Marcano and D. umbricola V. Marcano. Acantholichen dendroideus grows on mosses in shady and humid disturbed forest and is characterized by densely and dichotomously branched microsquamules, with a pruinose surface and large acanthohyphidia. Cyphellostereum mucuyense grows on mosses and sandy soils, lacks a jigsaw-puzzle-shaped hyphal sheath and prothallus, the fibrils are not penetrated by tubular fungal hyphae and the basidiocarp surface is finely arachnoid. This is the first record of Cyphellostereum in Venezuela. Dictyonema andinum might be confused with D. irrigatum (Berkeley & M. A. Curtis) Lücking from the Andaman Islands, but differs in the broader and ascending fibrils, a thicker photobiont layer and hypothallus, as well as the formation of stereoid-corticioid hymenophores. Dictyonema duidense from the Guayana Highlands and D. subsericeum from the Venezuelan Andes belong in the group of shelf-like morphotypes, with a semicircular thallus resembling D. sericeum s.str. Dictyonema duidense grows on tree trunks and is characterized by thick, very broad lobes and an areolate, rugose surface and it lacks clamp connections whereas D. subsericeum grows on tree branches and is characterized by variability of color (white, greenish blue, brown), size and shape (semicircular to hemispherical), filamentous lobes, a hirsute surface, fibrillose margins, clamp connections on the hyphae, cyphelloid hymenophores resembling attachment hapters, and very small, globose to subglobose basidiospores. Dictyonema laurae resembles D. subsericeum but is characterized by a continuous greenish blue crust-like thallus with an irregular surface, with discontinuous, fibrillose, and narrow margins projecting and ascending from the substrate, a hapteriform hymenophore, and its lacks clamp connections on the hyphae. Dictyonema subinvolutum is characterized by a jigsaw-puzzle-shaped hyphal sheath, fibrils penetrated by tubular fungal hyphae, and a smooth, pruinose, resupinate hymenophore. Finally, D. umbricola grows on mosses in very shady habitats and produces a corticioid hymenophore raised from the thallus, resembling apothecial discs, jigsaw-puzzle-shaped cells around the filaments and a distinct, discontinuous, white prothallus. Of the eight species of Dictyonema s.lat. registered from Venezuela, seven are known only from the Venezuelan Andes (A. dendroideus, C. mucuyense, D. andinum, D. laurae, D. subsericeum, D. subinvolutum, D. umbricola) and one is known only from the Venezuelan Amazonas (D. duidense). The high concentration of Acantholichen, Cyphellostereum and Dictyonema species in tropical America and especially of the latter genus in the Venezuelan Andes indicates that this region constitutes an important centre for speciation and endemism of Dictyonema clade/subtribe Dictyonematinae.

The Effect of Surrounding Vegetation on the Mycorrhizal Fungal Communities of the Temperate Tree Crataegus monogyna Jacq.

About 90% of all land plants form mycorrhiza to facilitate the acquisition of essential nutrients such as phosphorus, nitrogen, and sometimes carbon. Based on the morphology of the interaction and the identity of the interacting plants and fungi, four major mycorrhizal types have been distinguished: arbuscular mycorrhiza (AM), ectomycorrhizal (EcM), ericoid mycorrhiza, and orchid mycorrhiza. Although most plants are assumed to form only one type of mycorrhiza, some species simultaneously form associations with two mycorrhizal types within a single root system. However, the dual-mycorrhizal status of many species is under discussion and in some plant species the simultaneous association with two mycorrhizal types varies in space or time or depends on the ecological context. Here, we assessed the mycorrhizal communities associating with common hawthorn (Crataegus monogyna), a small tree that commonly associates with AM fungi, and investigated the potential factors that underlie variation in mycorrhizal community composition. Histological staining of C. monogyna roots showed the presence of a Hartig net and hyphal sheaths in and around the roots, demonstrating the capacity of C. monogyna to form EcM. Meta-barcoding of soil and root samples of C. monogyna collected in AM-dominated grassland vegetation and in mixed AM + EcM forest vegetation showed a much higher number of EcM sequences and OTUs in root and soil samples from mixed AM + EcM vegetation than in samples from pure AM vegetation. We conclude that C. monogyna is able to form both AM and EcM, but that the extent to which it does depends on the environmental context, i.e., the mycorrhizal type of the surrounding vegetation.

Amelioration of nitrate uptake under salt stress by ectomycorrhiza with and without a Hartig net.

Summary Salt stress is an important environmental cue impeding poplar nitrogen nutrition. Here, we characterized the impact of salinity on proton‐driven nitrate fluxes in ectomycorrhizal roots and the importance of a Hartig net for nitrate uptake.We employed two Paxillus involutus strains for root colonization: MAJ, which forms typical ectomycorrhizal structures (mantle and Hartig net), and NAU, colonizing roots with a thin, loose hyphal sheath. Fungus‐colonized and noncolonized Populus × canescens were exposed to sodium chloride and used to measure root surface pH, nitrate (NO 3 −) flux and transcription of NO3 − transporters (NRTs; PcNRT1.1, ‐1.2, ‐2.1), and plasmalemma proton ATPases (HAs; PcHA4, ‐8, ‐11). Paxillus colonization enhanced root NO 3 − uptake, decreased surface pH, and stimulated NRTs and HA4 of the host regardless the presence or absence of a Hartig net. Under salt stress, noncolonized roots exhibited strong net NO 3 − efflux, whereas beneficial effects of fungal colonization on surface pH and HAs prevented NO 3 − loss. Inhibition of HAs abolished NO 3 − influx under all conditions.We found that stimulation of HAs was crucial for the beneficial influence of ectomycorrhiza on NO 3 − uptake, whereas the presence of a Hartig net was not required for improved NO 3 − translocation. Mycorrhizas may contribute to host adaptation to salt‐affected environments by keeping up NO 3 − nutrition.

Deciphering interfungal relationships in the 410 million-year-old Rhynie chert: Glomoid spores under attack

Various types of other fungi colonize glomeromycotinan (Mucoromycota) spores in the Early Devonian Rhynie chert. However, relatively few of these associations have been described and evaluated in detail. One particular type of glomoid spore located in degrading land plant axes from the Rhynie chert provides evidence of (simultaneous) interaction with three different fungi. Massive callosities occur around the intrusion filaments of a chytrid-like parasite with epibiotic sporangia, while the hyphae of a delicate mycelial fungus extend into the spore lumen without triggering a recognizable host response. Several spores show large numbers of inwardly directed projections, which are regularly distributed and consist of a short hyphal branch encased in host wall material. The projections represent the penetration sites of a distinctive, mycelial fungus that forms a mantle-like hyphal sheath around the spores. This type of fungal interaction with glomeromycotinan spores has not previously been reported, and thus expands our knowledge of the numerous interfungal relationships that existed in early continental ecosystems.

New species ofDictyonemaandCyphellostereum(lichenizedBasidiomycota:Hygrophoraceae) from tropical Africa and the Indian Ocean, dedicated to the late Hildur Krog

Abstract: A small collection of lichenized fungi belonging in the Dictyonema clade from E Africa and Mauritius, tentatively identified as D. sericeum, revealed four species new to science, one in the genus Cyphellostereum and three in the genus Dictyonema s.str. Cyphellostereum bicolor from Mauritius is similar to C. imperfectum from Guatemata, but differs in the absence of a persistent white hypothallus and the less compact hyphal sheath of the fibrils, as well as the formation of resupinate hymenophores. Dictyonema album, also from Mauritius, belongs in the group of shelf-like morphotypes and is characterized within this group by very loose tufts dominated by sterile hyphae with very loosely arranged cyanobacterial fibrils. Dictyonema krogiae from Kenya is another new species in the group with shelf-like thalli and is the first species in this group known to form a distinctly layered thallus, with very compact photobiont layer, loose medulla, and more compact lower cortex. Finally, Dictyonema tricolor f...

Dictyonema huaorani(Agaricales: Hygrophoraceae), a new lichenized basidiomycete from Amazonian Ecuador with presumed hallucinogenic properties

Abstract Dictyonema huaorani, a new species represented by a well-developed specimen found in the Ecuadorian Amazon region, is described in this paper. The material was collected during a Harvard ethnobotanical expedition in 1981 and originally determined by Mason E. Hale Jr. as belonging in the genus Dictyonema (D. sericeum s.lat.) and possibly representing an undescribed species. The species is morphologically distinctive in forming densely woven, semicircular thalli, closely resembling those of the paleotropical D. ligulatum but lacking clamps and with hyphal sheath around the photobiont filaments that resembles those of Cyphellostereum species. The species was reported to have hallucinogenic properties and chemical analyses suggest certain substances present that are shared with the hallucinogenic mushroom Psilocybe cubensis. Due to our inability to use pure reference compounds and scarce amount of sample for compound identification, however, our analyses were not able to determine conclusively the pr...

Humic substances enhance growth and respiration in the basidiomycetes Trametes maxima under carbon limited conditions.

Humic substances (HS) represent the major reservoir of carbon (C) in ecosystems, and their turnover is crucial for understanding the global C cycle. Although basidiomycetes clearly have a role in HS degradation, much less is known about the effect of HS on fungal traits. We studied the alteration of physiological, biochemical, and morphological characteristics of Trametes maxima in the presence of HS. Both complete medium and minimal (C-limited) medium mimicking natural environmental conditions were used. Adding HS led to increased biomass yield, but under C-limited conditions the effect was more apparent. This result indicated that HS were used as an additional substrate and agreed with data showing a greater penetration of tritium-labeled HS into the cell interior under C-limited conditions. Humic substances induced ultra-structural changes in fungal cells, especially under C limitation, including reducing the thicknesses of the hyphal sheath and cell wall. In the minimal medium, cellular respiration increased nearly three-fold under HS application, while the corresponding effect in complete medium was lower. In addition, in the presence of inhibitors, HS stimulated either the cytochrome or the alternative pathway of respiration, depending on presence or absence of glucose in the medium. Our results suggest that, under conditions mimicking the natural environment, HS may play three major roles: as a surplus substrate for fungal growth, as a factor positively affecting cell morphology, and as an activator of physiological respiration.

Starting from scratch: Evolution of the lichen thallus in the basidiolichen Dictyonema (Agaricales: Hygrophoraceae)

Phylogenetic studies indicate that the basidiolichen genus Dictyonema s.lat., often thought to represent only a single genus with few species, includes several well-supported genus-level clades, all of which form associations with a unique lineage of obligately lichenized cyanobacteria (Rhizonema). In an attempt to elucidate the evolution and genus- and species-level diversification in Dictyonema s.lat., we generated 68 new sequences of the nuclear large subunit rDNA (nuLSU), the internal transcribed spacer (ITS), and the RNA polymerase II subunit (RPB2), for 29 species-level lineages representing all major clades of Dictyonema s.lat. and most of the species currently known. The multilocus phylogeny obtained via maximum likelihood and Bayesian approaches indicates the presence of five genus-level groups: a basal clade, Cyphellostereum, that is sister to the rest of the species, a paraphyletic grade representing Dictyonema s.str., and three clades representing the genera Acantholichen, Cora, and Corella. To determine the evolutionary transformations of the lichenized thallus in the group, ancestral character state reconstruction was done using six characters (lichenisation, thallus type, cortex type, hyphal sheath and haustorial type, photobiont morphology, and basidiocarp type). Our analysis indicates a progressive development of the lichenized thallus from loosely organized filamentous crusts with separate, cyphelloid basidiocarps in Cyphellostereum, to filamentous crusts with derived hyphal sheath and cyphelloid–stereoid basidiocarps partially incorporated into the lichen thallus in Dictyonema, to squamulose–foliose thalli with corticioid basidiocarps entirely supported by the lichen thallus in Cora. These results indicate a remarkable evolutionary integration of lichenized and reproductive tissues in Dictyonema s.lat., supporting the hypothesis that, at least in this case, lichenized thalli may have evolved from reproductive structures in their nonlichenized ancestors.

A first assessment of Galapagos basidiolichens

As part of an ongoing comprehensive inventory of Galapagos lichens, a first assessment of the morphology and anatomy of basidiolichens from the archipelago is presented here. It is the basis for further studies of the taxonomy, ecology and biogeography of this poorly known group of lichens. Four genera, all in Hygrophoraceae, can be distinguished: Acantholichen, Cora, Cyphellostereum and Dictyonema. Both Acantholichen and Cora are characterized by chroococcoid cyanobionts and a heteromerous thallus with a distinct upper cortex and photobiont layer. The monotypic Acantholichen pannarioides is entirely composed of small, branched, inflated squamules that appear densely pruinose because their cortical hyphae bear characteristically swollen, densely spinose end cells (acanthohyphidia); this species has never been observed fertile. The common Cora glabrata is foliose, forming large, radially zonate, conch-like, often tiled thalli, when fertile with circular lines of basidiocarps on its lower side. Dictyonema is distinguished by filamentous cyanobionts and distinctly filamentous thalli that are homomereous (i.e., not distinctly layered); all species of Dictyonema s.str. have trichomes (filamentose cyanobacterial photobionts) closely enveloped by fungal cells of a jigsaw pattern. In D. sericeum thallus filaments (i.e., individual fibrils) aggregate to form shelf-like structures similar in appearance to polyporoid bracket fungi; basidiocarps develop in irregular patches on the lower side of these shelves. In contrast, fibrils of D. schenkianum grow encrusting their substrate with irregularly to suberect trichomes, occasionally bearing basidiocarps dispersed across the thallus. Two other species in Galapagos show adpressed growth form and are described here as new: Dictyonema pectinatum, which is characterized by large parallel fibrils with paler, papillate tips, and D. galapagoense, characterized by thin trichomes of more squarrish elongate cells. The genus Cyphellostereum is represented by two species: the newly described C. imperfectum and an unnamed Cyphellostereum sp., both phenotypically similar to free-living cyanobacterial filaments. Cyphellostereum imperfectum has narrow photobiont filaments with irregular hyphal sheath leaving interspaces; macroscopically it shows a bluish green thallus with a distinct prothallus. Cyphellostereum sp. has a rather uncommon basidiolichen appearance: thin sctytonematoid fibrils surrounded by straight fungal cells forming shiny tufts. The new combination Cyphellostereum nitidum is also proposed. The ecology and taxonomy of Galapagos basidiolichens is briefly discussed and a key and short descriptions of all species are presented.

Cytology and ultrastructure of interactions between Ustilago esculenta and Zizania latifolia

Ustilago esculenta is a biotrophic smut fungus that parasitizes Zizania latifolia, an edible aquatic vegetable of the southern China region. Infection results in swelling of the upper parts of the Z. latifolia culm which are called jiaobai and have a unique flavor and delicacy and are popular among Chinese. The infection process of Z. latifolia by U. esculenta was investigated with light and electron microscopy. Distribution of hyphae was uneven in plants; hyphae were mainly present in the swollen upper parts (jiaobai), the nodal regions of mature culms and old rhizomes and buds or shoots. Hyphae were rare in the internodes of mature culms and were fewer in the internodes of old rhizomes. All new buds produced on the nodes of culms and rhizomes were infected by hyphae in November before and in March after overwintering. The hyphae grew into the buds from the parent nodes via intervascular tissues only or via parenchyma tissues and vascular bundles. Hyphae extended within and between the host cells and frequently formed hyphal aggregations or clusters, not only in the mature tissues but also in developing tissues. The typical interface between the fungal hyphal wall and invaginated host plasma membrane comprised a sheath. The sheath surrounding a hyphae comprised an outer electron-opaque matrix and an inner electron-dense layer. The electron-opaque matrix layers were thicker in jiaobai tissues, ranging from 0.28 to 0.85 μm. The electron-dense hyphal coatings were more conspicuous in the young buds or shoots and mature culms than in the jiaobai. The intercellular hyphae caused large cavity formation between the cells or rupture of host cell walls, for gaining entry into host cells. The broken host cell wall fused with the electron-opaque matrix of the hyphal sheath as an interactive interface. The teliospore wall and wall ornamentation development was the same in postmature jiaobai tissues with sporadic sori and in the huijiao (jiaobai tissues containing the massive sori), but a sheath enveloping the teliospore was more transparent in the process of teliospore development in the jiaobai than in the huijiao.

Spatial patterns in hyphal growth and substrate exploitation within norway spruce stems colonized by the pathogenic white-rot fungus Heterobasidion parviporum.

In Norway spruce, a fungistatic reaction zone with a high pH and enrichment of phenolics is formed in the sapwood facing heartwood colonized by the white-rot fungus Heterobasidion parviporum. Fungal penetration of the reaction zone eventually results in expansion of this xylem defense. To obtain information about mechanisms operating upon heartwood and reaction zone colonization by the pathogen, hyphal growth and wood degradation were investigated using real-time PCR, microscopy, and comparative wood density analysis of naturally colonized trees with extensive stem decay. The hyphae associated with delignified wood at stump level were devoid of any extracellular matrix, whereas incipient decay at the top of decay columns was characterized by a carbohydrate-rich hyphal sheath attaching hyphae to tracheid walls. The amount of pathogen DNA peaked in aniline wood, a narrow darkened tissue at the colony border apparently representing a compromised region of the reaction zone. Vigorous production of pathogen conidiophores occurred in this region. Colonization of aniline wood was characterized by hyphal growth within polyphenolic lumen deposits in tracheids and rays, and the hyphae were fully encased in a carbohydrate-rich extracellular matrix. Together, these data indicate that the interaction of the fungus with the reaction zone involves a local concentration of fungal biomass that forms an efficient translocation channel for nutrients. Finally, the enhanced production of the hyphal sheath may be instrumental in lateral expansion of the decay column beyond the reaction zone boundary.

Selective staining and visualization of hyphal sheath of a white-rot fungus Phanerochaete crassa WD1694 with phloxine B

The hyphal sheath is a morphological feature of many kinds of fungi. Although the fine structures of the sheath have been studied in detail by a number of electron microscopy techniques, the function and physiology of the hyphal sheath are not yet clarified. One reason for this is that the hyphal sheath is a colorless, mucilaginous, and delicate material so that it is not easily identified. We developed a simple method to visualize and identify the hyphal sheath of the white-rot fungus Phanerochaete crassa WD1694. The small mycelial pellets in shaken liquid cultures of P. crassa WD1694 were stained directly with phloxine B. Both the hyphae and the hyphal sheath that filled the gaps between each of the hyphae were visualized and observed by light microscopy. The stained hyphae were further studied by transmission electron microscopy, atomic force microscopy, and fl uorescence microscopy. Based on these observations, we confirmed that the staining of the hyphae was also due to the presence of the hyphal sheath that closely covered the fungal cell wall. These results clearly showed that the hyphal sheath was selectively stained with phloxine B and could be observed and identified by conventional light microscopy.

Nitrogen fixation associated with Suillus tomentosus tuberculate ectomycorrhizae on Pinus contorta var. latifolia.

Tuberculate ectomycorrhizae are a unique form of ectomycorrhiza where densely packed clusters of mycorrhizal root tips are enveloped by a thick hyphal sheath to form a tubercle. The functional significance of such a unique structure has not previously been established. The purpose of the present study was to investigate and measure the potential nitrogenase activity associated with Suillus tomentosus/Pinus contorta tuberculate ectomycorrhizae in two stand ages, young and old, and across a range of nitrogen-poor soil conditions. Short roots were compared with other mycorrhizae and non-mycorrhizal secondary roots using tuberculate ectomycorrhizae. Assessment of nitrogenase activity was determined and quantitative measurements were taken on tuberculate ectomycorrhizae in situ in a variety of different circumstances, by using an adaptation of the acetylene reduction assay. Significant nitrogenase activity was measured associated with S. tomentosus/P. contorta tuberculate ectomycorrhizae whereas no nitrogenase activity was measured with non-tuberculate mycorrhizae or secondary roots without mycorrhizae. Average nitrogenase activity ranged from undetectable to 5696.7 nmol C2H4 g(-1) tubercle 24 h(-1). Maximum nitrogenase activity was 25,098.8 nmol C2H4 g(-1) tubercle 24 h(-1). Nitrogenase activity was significantly higher in young stands than in old stands of P. contorta. Season or some covariate also seemed to affect nitrogenase activity and there was suggestion of a site effect. Suillus tomentosus/P. contorta tuberculate ectomycorrhizae are sites of significant nitrogenase activity. The nitrogenase activity measured could be an important contribution to the nitrogen budget of P. contorta stands. Season and stand age affect levels of nitrogenase activity.

Morphological and molecular evidence supporting an arbutoid mycorrhizal relationship in the Costa Ricanpáramo

This study examines evidence for a particular arbutoid mycorrhizal interaction in páramo, a high-altitude neotropical ecosystem important in hydrological regulation but poorly known in terms of its fungal communities. Comarostaphylis arbutoides Lindley (Ericaceae) often forms dense thickets in Central American páramo habitats. Based on phylogenetic classification, it has been suggested that C. arbutoides forms arbutoid mycorrhizae with diverse Basidiomycetes and Ascomycetes; however, this assumption has not previously been confirmed. Based on field data, we hypothesized an arbutoid mycorrhizal association between C. arbutoides and the recently described bolete Leccinum monticola Halling & G.M. Mueller; in this study, we applied a rigorous approach using anatomical and molecular data to examine evidence for such an association. We examined root samples collected beneath L. monticola basidiomes for mycorrhizal structures, and we also compared rDNA internal transcribed spacer (ITS) sequences between mycorrhizal root tips and leaf or basidiome material of the suspected symbionts. Root cross sections showed a thin hyphal sheath and intracellular hyphal coils typical of arbutoid mycorrhizae. DNA sequence comparisons confirmed the identity of C. arbutoides and L. monticola as the mycorrhizal symbionts. In addition, this paper provides additional evidence for the widespread presence of minisatellite-like inserts in the ITS1 spacer in Leccinum species (including a characterization of the insert in L. monticola) and reports the use of an angiosperm-specific ITS primer pair useful for amplifying plant DNA from mycorrhizal roots without co-amplifying fungal DNA.

Anatomy and ultrastructure of mycorrhizal associations of neotropical Ericaceae

Ericaceae are obligatory associated with symbiotic fungi forming several, distinctive categories of mycorrhizas. While ericoid, arbutoid, and monotropoid mycorrhizas are known since many years from ericads of the northern hemisphere and the ericoid mycorrhiza also from Australia, a further mycorrhizal category with hyphal sheath, Hartig net, and intracellular colonization was described by us recently and termed cavendishioid mycorrhiza because it was found on Cavendishia nobilis, a species belonging to the Andean clade (Vaccinioideae) of Ericaceae. As the previous findings indicated a correlation between the mycorrhizal category and the systematic position of Ericaceae, we tested the hypothesis that other ericads of the Andean clade might also form cavendishioid mycorrhizas, while ericads occurring in the same area but not belonging to the Andean clade might not. Mycorrhizas of 20 different ericaceous species, 15 belonging to the Andean clade and 5 to other Vaccinioideae or Ericoideae, were sampled in the tropical mountain rain forest area of South Ecuador and investigated by light and electron microscopy. All the 15 members of the Andean clade ericads displayed a hyphal sheath, as well as inter- and intracellular colonization by hyphae as was found on Cavendishia previously. The five species not belonging to the Andean clade ericads displayed only intracellular colonization by hyphae and hence were typical ericoid mycorrhizal. Ultrastructural studies revealed Sebacinales and ascomycetes as mycorrhiza formers in both associations even within one single cell. The results thus support the hypothesis that the Andean clade of Ericaceae forms mycorrhizas distinct from the arbutoid category and most likely presents an independent evolutionary line in the Ericaceae derived from the ericoid mycorrhizas, justifying the new term “cavendishioid mycorrhiza”.

Fungal relationships and structural identity of their ectomycorrhizae

Aproximately 5,000–6,000 fungal species form ectomyorrhizae (ECM), the symbiotic organs with roots of predominantly trees. The contributing fungi are not evenly distributed over the system of fungi. Within Basidiomycota exclusively Hymenomycetes and within Ascomycota exclusively Ascomycetes contribute to the symbiosis. Hymenomycetes play a big part, Ascomycetes a minor role; Zygomycetes only form exceptionally ECM. Responsible for ascomycetous ECM are mostly Pezizales with their hypogeous derivatives, whereas Boletales, Gomphales, Thelephorales, Amanitaceae, Cantharellaceae, Cortinariaceae, Russulaceae, and Tricholomataceae are the most important ectomycorrhizal relationships within Hymenomycetes. ECM, as transmitting organs between soil and roots, are transporting carbohydrates for growth of mycelium and fruitbodies from roots and have to satisfy the tree’s demand for water and nutrients. The latter task particularly influences the structure of ECM as nutrients are patchily distributed in the soil and saprotrophic as well as ectomycorrhizal fungi can act as strong competitors for nutrients. In focusing these requirements, ECM developed variously structured hyphal sheaths around the roots, the so-called mantles, and differently organized mycelium that emanates from the mantle, the so-called extramatrical mycelium. The mantles can be plectenchymatous consisting of loosely woven, differently arranged hyphae or they are densely packed, forming a pseudoparenchyma similar to the epidermis of leaves. The extramatrical mycelium grows either as simple scattered hyphae from the mantle into the soil or it can be united to undifferentiated rhizomorphs with a small reach or to highly organized root-like organs with vessel-like hyphae for efficient water and nutrient transport from distances of decimeters. Cystidia, sterile and variously shaped hyphal ends, possibly appropriate for preventing animal attack, in addition, can cover mantles and rhizomorphs. Although only a limited number of species could be considered, some general conclusions are possible.

Sebacinales form ectendomycorrhizas with Cavendishia nobilis, a member of the Andean clade of Ericaceae, in the mountain rain forest of southern Ecuador

Cavendishia nobilis var. capitata is an endemic member of the Ericaceae growing as a hemiepiphyte in the tropical mountain rain forest of southern Ecuador. Mycorrhizas were collected from 20 individuals along an altitudinal gradient between 1850 and 2300 m. Transmission electron microscopy was used to study the symbiotic association in detail, and phylogenetic analyses based on nuclear rDNA coding for the ribosomal large subunit (nucLSU) were carried out to identify the associated mycorrhizal fungi. Microscopic and ultrastructural investigations showed the formation of a hyphal sheath, intercellular penetration of fine hyphae and colonization of the cortical cells by swollen hyphae of the same fungus. These structures were formed by hymenomycetes and ascomycetes. Molecular phylogenetic analysis detected seven groups of mycorrhizal fungi belonging to the Sebacinales. This is the first study to obtain evidence of ectendomycorrhizas in the Vaccinioideae. The ascomycetous nucLSU sequences belonged to members of the Leotiomycetes. The ectendomycorrhiza of C. nobilis with Sebacinales is discussed as a specific, hitherto undescribed mycorrhizal subcategory of ectomycorrhizas. We propose the term 'cavendishioid mycorrhiza'. This subcategory is most likely specific for the Andean clade of Ericaceae.

Colonization with Hebeloma crustuliniforme increases water conductance and limits shoot sodium uptake in white spruce ( Picea glauca ) seedlings

White spruce [Picea glauca (Moench) Voss] seedlings were inoculated with Hebeloma crustuliniforme and treated with 25 mM NaCl to examine the effects of salinized soil and mycorrhizae on root hydraulic conductance and growth. Mycorrhizal seedlings had significantly greater shoot and root dry weights, number of lateral branches and chlorophyll content than non-mycorrhizal seedlings. Salt treatment reduced seedling growth in both non-mycorrhizal and mycorrhizal seedlings. However, needles of salt-treated mycorrhizal seedlings had several-fold higher needle chlorophyll content than that in non-mycorrhizal seedlings treated with salt. Mycorrhizae increased N and P concentrations in seedlings. Na levels in shoots and roots of salt-treated mycorrhizal seedlings were significantly lower and root hydraulic conductance was several-fold higher than in non-mycorrhizal seedlings. A reduction of about 50% in root hydraulic conductance of mycorrhizal seedlings was observed after removal of the fungal hyphal sheath. Transpiration and root respiration rates were reduced by salt treatments in both groups of seedlings compared with the controls, however, both transpiration and respiration rates of salt-treated mycorrhizal seedlings were as high as those in the non-mycorrhizal seedlings that had not been subjected to salt treatment. The reduction of shoot Na uptake while increasing N and P absorption and maintaining high transpiration rates and root hydraulic conductance may be important resistance mechanisms in ectomycorrhizal plants growing in salinized soil.

Carbon dioxide concentration and nitrogen input affect the C and N storage pools in Amanita muscaria-Picea abies mycorrhizae.

We studied the influence of elevated atmospheric CO2 concentration ([CO2]) on the vacuolar storage pool of nitrogen-containing compounds and on the glycogen pool in the hyphal sheath of Amanita muscaria (L. ex Fr.) Hooker-Picea abies L. Karst. mycorrhizae grown with two concentrations of ammonium in the substrate. Mycorrhizal seedlings were grown in petri dishes on agar containing 5.3 or 53 mg N l(-1) and exposed to 350 or 700 microl CO2 l(-1) for 5 or 7 weeks, respectively. Numbers and area of nitrogen-containing bodies in the vacuoles of the mycorrhizal fungus were determined by light microscopy linked to an image analysis system. The relative concentration of nitrogen in the vacuolar bodies was measured by electron energy loss spectroscopy (EELS). Glycogen stored in the cytosol was determined at the ultrastructural level by image analysis after staining the sections (PATAg test). Shoot dry weight, net photosynthesis and relative amounts of N in vacuolar bodies were greater at the higher N and CO2 concentrations. The numbers and areas of vacuolar N-containing bodies were significantly greater at the higher N concentration only at ambient [CO2]. In the same treatment the percentage of hyphae containing glycogen declined to nearly zero. We conclude that, in the high N/low [CO2] treatment, the mycorrhizal fungus had an insufficient carbohydrate supply, partly because of increased amino acid synthesis by the non-mycorrhizal rootlets. When [CO2] was increased, the equilibrium between storage of glycogen and N-containing compounds was reestablished.

Observations on the biology and ultrastructure of the asci and ascospores of Julella avicenniae from Malaysia

Ultrastructure of the marine, bitunicate, ascomycete Julella avicenniae is presented and compared with the marine Pleospora gaudefroyi. Asci of J. avicenniae possess an ocular chamber, a thick endoascus, and a thinner ectoascus. Pseudoparaphyses are enveloped by mucilage (hyphal sheath) which stains with ruthenium red. The mucilage appears to be an extension of the pseudoparaphysis cell wall and internally these cells contain an array of vesicles. Muriform ascospores are surrounded by an exosporial sheath, an electrondense episporium and a bilamellate mesosporium. Optimum conditions for growth are 25–30 °C in 100% artificial seawater glucoseyeast extract-tryptone media, but the fungus also is able to grow at 35° and at higher salinities. The ability of the fungus to withstand extremes of environmental conditions is discussed.