Conservation of biotrophy in Hygrophoraceae inferred from combined stable isotope and phylogenetic analyses

The nutritional modes of genera in Hygrophoraceae (Basidiomycota: Agaricales), apart from the ectomycorrhizal Hygrophorus and lichen-forming taxa, are uncertain. New δ(15)N and δ(13)C values were obtained from 15 taxa under Hygrophoraceae collected in central Massachusetts and combined with isotopic datasets from five prior studies including a further 12 species using a data standardization method to allow cross-site comparison. Based on these data, we inferred the probable nutritional modes for species of Hygrophorus, Hygrocybe, Humidicutis, Cuphophyllus and Gliophorus. A phylogeny of Hygrophoraceae was constructed by maximum likelihood analysis of nuclear ribosomal 28S and 5.8S sequences and standardized δ(15)N and δ(13)C values were used for parsimony optimization on this phylogeny. Our results supported a mode of biotrophy in Hygrocybe, Humidicutis, Cuphophyllus and Gliophorus quantitatively unlike that in more than 450 other fungal taxa sampled in the present and prior studies. Parsimony optimization of stable isotope data suggests moderate conservation of nutritional strategies in Hygrophoraceae and a single switch to a predominantly ectomycorrhizal life strategy in the lineage leading to Hygrophorus. We conclude that Hygrophoraceae of previously unknown nutritional status are unlikely to be saprotrophs and are probably in symbiosis with bryophytes or other understory plants.     

Hidden diversity uncovered in Hygrophorus sect. Aurei (Hygrophoraceae), including the Mediterranean H. meridionalis and the North American H. boyeri, spp. nov.

For many years, the binomial Hygrophorus hypothejus was widely applied to collections from various geographical regions in different continents, assuming a circum-boreal and circum-mediterranean distribution for this species. This hypothesis, however, had never been put to the test. To assess the diversity and species-limits within this complex of yellow-coloured waxcaps, a phylogenetic, morphological and taxonomical investigation into Hygrophorus sect. Aurei and similar species in sect. Olivaceoumbrini was carried out, including material of pan-European origin, as well as the east and west coasts of North America. Following sequencing of the ITS rDNA locus, nine lineages are confirmed in sect. Aurei, most of them highly continentalised. Of these, two are new to science, introduced here as Hygrophorus boyeri sp. nov., from Pinus banksiana and P. rigida forests in eastern North America and from P. muricata and P. contorta forests in western North America, and Hygrophorus meridionalis sp. nov., from Pinus brutia and Pinus halepensis forests in the island of Cyprus and mainland Greece. H. hypothejus is lectotypified and epitypified, and here resolved as a strictly European species, with the old forgotten taxon Hygrophorus siccipes revived as its North American vicariant. The placement of Hygrophorus fuligineus in sect. Aurei is phylogenetically confirmed and detailed comparisons between morphologically similar and phylogenetically affiliated taxa in sect. Aurei and sect. Olivaceoumbrini are provided. The chronic confusion associated with Hygrophorus fuscoalbus, a highly controversial taxon described from Germany nearly two centuries ago and variously interpreted since, is discussed, concluding that this name is too ambiguous to be applied to any currently recognized species.     

Indole and 3-chloroindole: The source of the disagreeable odor of Hygrophorus paupertinus

The odor emanating from sporocarps of Hygrophorus paupertinus is disagreeable and fecal-like. Solid phase microextraction (SPME) and analysis by gas chromatography-mass spectrometry (GC-MS) showed 1-octen-3-ol, indole and 3-chloroindole were responsible for the odor. This is the first case in which 3-chloroindole has been identified from a terrestrial organism.     

Hygrophorus yukishiro sp. nov., a new vernal edible mushroom from Nagano Prefecture,Japan

A novel basidiomycetous species, Hygrophorus yukishiro, is described from Japan. The basidiomata of this fungus have been referred to as “yuki-shiro”, “yuki-take”, and “yuki-wari”, and used as an edible mushroom in the central region of Nagano, Japan. Hygrophorus yukishiro fruits in the early spring season under oak-dominated stands. Morphological and phylogenetic analyses of the internal transcribed spacer region of the nuclear ribosomal RNA gene indicated that H. yukishiro is a sister species of H. arbustivus and belongs to section Hygrophorus, subsect. Fulventes. Hygrophorus yukishiro is similar in its external morphology and fruiting phenology to H. roseobrunneus, H. leucophaeo-ilicis, and H. roseodiscoideus, but differs in the color and texture of its pileus surface and microscopic characteristics.     

Purification and characterization of an anti-(A+B) specific lectin from the mushroom Hygrophorus hypothejus.

A lectin (HHL) was isolated from the fruiting body of the mushroom Hygrophorus hypothejus by a combination of affinity chromatography on stromas of group B erythrocytes embedded in polyacrylamide gel, and DEAE-trisacryl and gel filtration chromatography. Its molecular mass, as determined by gel filtration, is estimated to be 68000 kDa and its structure is tetrameric with four identical subunits assembled with non-covalent bonds. HHL agglutinates specifically A and B blood group erythrocytes and in hemagglutination inhibition assays, exhibits sugar-binding specificity toward lactose, the anomeric alpha form being more effective than the beta form.     

Asexual reproduction of <Emphasis Type="Italic">Hygrophorus olivaceoalbus</Emphasis> by intracellular microsclerotia in root cells of <Emphasis Type="Italic">Picea abies</Emphasis> – A winner of ozone stress?

Hygrophorus olivaceoalbus has long been known as an ectomycorrhizal fungus, formerly designated with the artificial binomen Piceirhiza gelatinosa. Recently, it has been found to be abundant and very frequent under double ambient ozone free air fumigation of mature Norway spruce trees. As it has already been reported that this fungus can form intercellular hyphae within the root meristem, a more detailed study was performed to clarify its type of root colonization. The present study not only revealed intercellular hyphae within the meristematic zone but also intracellular hyphae within root cortex cells which grow towards and into large tannin droplets—phenolic compounds usually deposited as defensive aids—to finally fill them completely. The hyphal assemblages become globular which at first sit as separated hyphal balls within cells still containing cytoplasm. Later on, they are apparently released from the root cells, presumably as microsclerotia for dispersal of the species. Old ectomycorrhizae (ECM) show an apical pore, and later a large orifice of a tube-like cylinder formed by the thick, persistent hyphal mantle. The root tissue is progressively degrading towards proximal parts. Disintegrating root cells apparently liberate the microsclerotia through the orifice. Further studies have to find out the mechanism by which the microsclerotia are liberated and whether they operate as asexual propagules and lastly how and by whom they are propagated. As these ECM are found under ozone stress, and with identical features at higher altitudes, stress impact on trees might be the causative agent for the high frequency and abundance of Hygrophorus olivaceoalbus ECM.     

The phylogenetic position of Haasiella (Basidiomycota, Agaricomycetes) and the relationships between H. venustissima and H. splendidissima

Based on a combined ITS-LSU rDNA analysis, the omphalinoid genus Haasiella is shown to be part of the Hygrophoroid clade and sister to Hygrophorus. H. venustissima and H. splendidissima are conspecific; H. splendidissima is the tetrasporic/clamped and heterothallic form of H. venustissima.     

Coenological research on macrofungi in evergreen oak woods in the hills near Siena (Italy)

For a mycocoenological study of macromycetes in Mediterranean evergreen forests in the Siencese province, five stands belonging to theQuercion ilicis have been observed during more than two years. The phytosociological analysis of these evergreen oak woods, situated at the limit of the geographical distribution of the alliance, reveals them as intermediate between the typical Mediterranean vegetation and the sub-Mediterranean deciduous broad-leaved forests. 181 fungal species were recognized and assigned to seven ecological groups on the basis of substratum, habitat, forest management and/or particular mycorrhizal relationship. Our results have been compared with those obtained in some European deciduous forests. Such a comparison shows the occurrence of 16 thermophilous fungal species in the Siena woods which are missing or very rare in the European deciduous forests. On the basis of our present knowledge, five of these species (Boletus lepidus, Hygrophorus dichrous, Hygrocybe nigrescens, Inocybe similis, Phellinus torulosus) may be considered as strictly tied to the evergreen oak woods. Five others, which have been reported for several European phytocoenoses, seem to be ‘preferential taxa’ of our evergreen oak woods, namely:cortinarius calochrous, C. sodagnitus, Hygrophorus arbustivus, H. russula, Lyophyllum immundum. On the whole the highest similarities have been found with the more xerothermophilous European forests (Querco-Lithospermetum andSorbo-Quercetum). The presence in theQuercus ilex woods of numerous more mesophilous fungi, some of which are widespread in beech forests, underlines the transitional nature of the phytocoenoses studied.     

Hygrophorones A-G: fungicidal cyclopentenones from Hygrophorus species (Basidiomycetes)

Twenty new 5-(hydroxyalkyl)-2-cyclopentenone derivatives (hygrophorones) could be isolated from Hygrophorus latitabundus, H. olivaceoalbus, H. persoonii, and H. pustulatus. Their fungicidal activity was exemplarily tested. The hygrophorones have structural similarities to the antibiotic pentenomycin. Chemically, hygrophorones are 2-cyclopentenones with hydroxy or acetoxy substituents at C-4 and/or C-5. An odd-numbered 1' oxidized alkyl chain (C(11), C(13), C(15), or C(17)) is attached at C-5. In addition, from H. persoonii the new gamma-butyrolactone derivative [5-(E)-2-hydroxytetradexylidene-5H-furan-2-one] could be isolated. Some hygrophorones are responsible for the color reaction of the stipes of these fungi upon treatment with potassium hydroxide solution. Structural elucidations are based on 1D ((1)H, (13)C) and 2D (COSY, NOESY, HSQC, HMBC) NMR spectroscopic analyses as well as HR-FT-ICR-MS investigations.     

中国蜡伞科已知种类及其分布

文中对中国蜡伞科(Hygrophoraceae)真菌进行了整理、修订,简要介绍了其生境和分布,并对该科的分类学问题进行了讨论。重点整理了文献记载该科的102个种,广泛分布于28个省区。其中有效名称86个,包括蜡伞属(Hygrophorus)47个,湿伞属(Hygrocybe)36个,湿皮伞属(Humidicutis)1个,坛杯伞属(Ampulloditocybe)1个,金盖菇属(Chrysomphalina)1个,应转移到其他科的有5种,错拼名称有9种,另有存疑种2种。     

吉林省担子菌补记(七)

报道了吉林省新记录分布担子菌6种,即肉褐鳞环柄菇(Lepiota brunneoincarnataChodat&C.Martín)、短柄金舌囊蘑[Melanoleuca brevipes(Bull.)Pat]、紫皮丽蘑[Calocybe ionides(Bull.)Donk]、皮尔松湿伞(HygrophoruspersooniiArnolds)、微黄拟锁瑚菌[Clavulinopsis helvola(Pers.)]和弹球菌(Sphaerobolus stellatusTode)。其中,弹球菌属为吉林省新记录属。凭证标本存放于吉林农业大学菌物标本馆(HMJAU)。     

Diversity within the <Emphasis Type="Italic">Hygrophorus agathosmus</Emphasis> group (Basidiomycota,Agaricales) in Northern Europe

The North European species of the Hygrophorus agathosmus group in subsection Tephroleuci were studied. Three new species are identified based on morphology, ecology and sequence data. Two species are associated with Pinus spp. One of these is described here as H. suaveolens, while the other one is only known from one locality in the Nordic countries and seems to have a more South European distribution range. A closely related sister species to H. agathosmus is identified based on ITS sequence data, H. cf. agathosmus. It is confirmed to have an intercontinental distribution range and to be associated with Picea spp. probably on more acidic to neutral soil, whereas H. agathosmus s.s. has a more limited North-East European distribution range and occurs in older and rich Picea abies forests. A neotype for H. agathosmus is here selected from South Sweden. Hygrophorus agathosmus f. albus and H. agathosmus f. aureofloccosus are confirmed as forms. No genetic differences in the ITS region between specimens with grey cap colour and the two forms were observed. A key to the species in Northern Europe is provided.     

Effects of acid irrigation and liming on the production of fruit bodies by ectomycorrhizal fungi

The aim of this work was to examine the effects of an increase in acid rain upon community diversity and productivity of the ectomycorrhizal fungi in a Norway spruce stand and the possible amelioratory effects of liming. No obvious adverse effects on either the diversity or productivity were found. Acid irrigation appeared to enhance the fruiting of Russula ochroleuca, as did irrigation with normal rain. A combination of acid irrigation and liming resulted in a large increase in the fruiting of Hygrophorus pustulatus. Thirty times more fruit bodies were recorded from this treatment than from the control. A principal component analysis of the fruit body data suggested that soil organic horizon pH may be an important determinant of ectomycorrhizal community structure within the spruce stand.     

Zur Taxonomie,Verbreitung und Ökologie des Filzigen Schmierlings Chroogomphus helveticus (Sing.) Moser

Chroogomphus helveticus is an european, dealtomontanous species with a south-hercynian-alpian-carpathian and illyrian-macedonian distribution. It grows in association with various conifers. Picea abies, Pinus cembra, Pinus peuce, Pinus strobus and Abies alba probably are partners of the mycorrhiza. The area is similar to that of Abies alba, Hygrophorus marzuolus and various dealpine plants. Distribution centres are districts, where 5-needled pines (Pinus cembra, Pinus peuce) are to be found naturally. Chroogomphus helveticus is to be found in an altitude from 300 to over 2000 m NN. The species is acidophilous, but not strongly bound to acid soils. A subdivion of the species into subspecific taxa is not possible. The closest relative of Chroogomphus helveticus occurs in Asia and Northern America. An other related european species is only known from England.     

Candida siamensis sp. nov., an anamorphic yeast species in the Saturnispora clade isolated in Thailand

Three strains of a novel anamorphic yeast species were isolated from natural samples collected in Thailand, including strain EF10 isolated from sediment in a mangrove forest, strain ST-473 from rotten wood, and strain ST-479 from a wild mushroom ( Hygrophorus sp.). Analysis of the D1/D2 domains of the large-subunit rRNA gene sequence revealed that the sequence of strain EF10 differed from the other two strains (ST-473 and ST-479), which had identical sequences, by only one nucleotide (nt) substitution in 535 nt. The closest species in terms of pairwise sequence similarity was Saturnispora mendoncae , with 32 nt substitutions and seven gaps in 535 nt. Phylogenetic analysis demonstrated that the three strains form a cluster with S. mendoncae and the other eight species in the Saturnispora clade. Phenotypic characteristics of the three strains were the same including proliferation by multilateral budding, absence of ascospores and negative diazonium blue B and urease reactions. The major ubiquinone was Q-7. On this basis, the three strains were assigned to a single novel species of the genus Candida , for which the name Candida siamensis sp. nov. is proposed. The type strain is EF10^T (=BCC 29901^T=NBRC 104878^T=CBS 11022^T).     

Identification of Picea-ectomycorrhizae by comparing DNA-sequences

Identification of the fungi forming ectomycorrhiza is still a great challenge. Ectomycorrhizae of Picea abies, collected in southwest Germany during several years and described as morphotypes, were identified using LSU and ITS sequences. To this the fungal sequences amplified from the mycorrhizae were compared with sequences from identified sporocarps. The fungal partner of Piceirhiza gelatinosa was identified as Hygrophorus olivaceoalbus, the fungal partner of Piceirhiza rosa-nigrescens was identified as Dermocybe cf. semisanguinea, and the fungal partner of a white mycorrhiza, described here for the first time, belongs to the Hebeloma velutipes group (Hebeloma crustuliniforme complex). Identification to genus level was possible for Piceirhiza lanuginosa where a Cortinarius-species is the fungal partner. A Tomentella-species forms a newly described light-brown mycorrhiza. Identification to family and to order-level was achieved for a milky-dull silvery mycorrhiza (Cortinariaceae), and Piceirhiza globulifera (Aphyllophorales), respectively. Ten samples of white, slightly bent mycorrhizae were formed by 8 different Cortinarius-species, including among others Cortinarius traganus, C. delibutus, and C. brunneus. The fungal partners of several brown, smooth mycorrhizae displaying only a Hartig net are Ascomycetes, among these are Wilcoxina cf. mikolae and Hymenoscyphus cf. ericae.     

Mycorrhizae in Adenostoma fasciculatum Hook. & Arn.: a combination of unusual ecto- and endo-forms

 The mycorrhizal status of Adenostoma fasciculatum, the dominant shrub in California chaparral, has been unclear. In two typical, nearly monospecificstands, A. fasciculatum was found to have arbuscules and intercellular hyphae. Antisera detected hyphae of the arbuscular mycorrhizal (AM) fungal genera Acaulospora, Glomus, and Scutellospora, although we found only spores of Glomus. Some roots had partial sheaths and inter- and intracellular septate fungi without indications of root necrosis. Ectomycorrhizal root tips were also found, including Cenococcum and other unknown taxa. Sporocarps of EM fungi including species of Rhizopogon, Pisolithus, Balsamia, Laccaria, Hygrophorus, and Cortinarius were found in the stand, with no other EM or arbutoid mycorrhizal plants nearby. These observations indicate that A. fasciculatum forms mycorrhizae with AM, septate, and EM fungi, but often fails to form easily recognizable mycorrhizal structures. Accepted: 5 September 1998     

Microflora of a coniferous forest of the Mexican basin

Summary Soil profiles were made at three locations corresponding to the three principal coniferous groups:Pinus montezumae, Abies religiosa andPinus hartwegii. The soil samples were submitted to microbiological, physical and chemical analysis. Mycorrhizic fungi were collected throughout this zone and 25 species of ectotrophic and 1 facultative mycorrhizic fungiLaccaria laccata were found.Lycoperdon perlatum andHygrophorus chrysodon were in the three communities, as wellAmanita muscaria, Boletus granulatus andCantharellus cibarius were only in the Pinus communities. The experimental condition used in order to prove symbiosis showed us several symbionts in the root for each one of the three different conifers. As regards general microflora, the greatest number of bacteria and fungi was present in the surface horizon of Abies soil; these soils are deep and moist throughout the year and with a good drainage, so that seasonal variations have little or no effect on soil environmental conditions and that favors the microbiological development. In the soils of the Pinus communities the number of actinomycetes and of bacteria is higher immediately below the surface horizon.     

贵州梵净山大型真菌生态分布及其资源评价

本文报道了梵净山的大型真菌365种,分属于124属、41科。其中食用菌105种,药用菌87种,毒菌33种,木腐菌131种及菌根菌114种。梵净山的大型真菌分属于下列林型和植被型:阔叶林型;针阔混交林型;针叶林型;灌丛型和荒地型。营养方式主要分为寄生、腐生和共生三大类群。     

Mannose-specific lectin from the mushroom Hygrophorus russula

A lectin was purified from the mushroom Hygrophorus russula by affinity chromatography on a Sephadex G-50 column and BioAssist S cation exchange chromatography and designated H. russula lectin (HRL). The results of sodium dodecyl sulfate-polyaclylamidegel electrophoresis, gel filtration and matrix-assisted laser desorption ionization time-of-flight mass spectrometry of HRL indicated that it was composed of four identical 18.5?kDa subunits with no S-S linkage. Isoelectric focusing of the lectin showed bands near pI 6.40. The complete sequence of 175 amino acid residues was determined by amino acid sequencing of intact or enzyme-digested HRL. The sequence showed homology with Grifola frondosa lectin. The cDNA of HRL was cloned from RNA extracted from the mushroom. The open reading frame of the cDNA consisted of 528?bp encoding 176 amino acids. In hemagglutination inhibition assay, α1-6 mannobiose was the strongest inhibitor and isomaltose, Glcα1-6Glc, was the second strongest one, among mono- and oligosaccharides tested. Frontal affinity chromatography indicated that HRL had the highest affinity for Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAc, and non-reducing terminal Manα1-6 was essential for the binding of HRL to carbohydrate chains. The sugar-binding specificity of HRL was also analyzed by using BIAcore. The result from the analysis exhibited positive correlations with that of the hemagglutination inhibition assay. All the results suggested that HRL recognized the α1-6 linkage of mannose and glucose, especially the Manα1-6 bond. HRL showed a mitogenic activity against spleen lymph cells of an F344 rat. Furthermore, an enzyme-linked immunosorbent assay showed strong binding of HRL to human immunodeficiency virus type-1 gp120.