Isolation and characterization of developmental variants in fruiting using a homokaryotic fruiting strain ofCoprinus cinereus

Developmental variants in fruiting ofCoprinus cinereus were induced by mutagenizing oidia of the homokaryotic fruiting strain CopD5-12 with UV light. Through screening of 2,696 isolates, 1,018 strains exhibited defects in fruiting and were classified into 8 groups: (1) knotless variants, which fail to form hyphal knots, the first visible sign of fruiting; (2) primordiumless variants, which form hyphal knots but fail to develop fruit-body primordia; (3) maturationless variants, which form fruit-body primordia but do not form mature fruit bodies; (4) elongationless variants, which form mature fruit bodies with short stipes; (5) expansionless variants, which form mature fruit bodies with unexpanded pilei; (6) sporeless variants, which fail to produce black basidiospores, resulting in fruit bodies with white pilei after maturation; (7) compound type, which includes variants exhibiting several of the phenotypes described above; (8) others, including variants that produce a “dark stipe” even under in light/dark conditions, which is formed under continuous darkness in the wild-type. Two elongationless variants were characterized histologically.     

A new species of <Emphasis Type="Italic">Cortinarius</Emphasis> sect. <Emphasis Type="Italic">Orellani</Emphasis> from Japan

Cortinarius breviradicatus sp. nov., found in deciduous forests, is described and illustrated from Niigata, Japan. It is characterized by its medium-sized to large dark brown basidiocarp, acutely conical pileus, and rooting stipe, and by subglobose to broadly ellipsoid basidiospores. In addition, the extracting solution from its basidiocarps exhibits a strong fluorescence around 400–430 nm in ultraviolet radiation (250 nm), which was observed in a species of Cortinarius sect. Orellani. The new species belongs to the section Orellani. The differences between the new taxon and similar species are briefly discussed.     

Sequential production of enzymes and basidiospore formation in fruiting bodies of Coprinus macrorhizus

The production of NADP-dependent glutamate dehydrogenase was initiated at the stage of first meiotic prophase in pileus cells but not in stipe cells of dikaryotic and monokaryotic fruiting bodies in Coprinus macrorhizus. The production of chitinase and glucanases assayed with laminarin and lichenan was observed after the completion of meiosis only in pileus cells. The light conditions that were effective for the delay or inhibition of cellular events in the pileus cells were also effective for the delay or inhibition of enzyme production. But all sporeless mutants tested, which were defective at the various stages of basidiospore formation, produced the normal levels of these enzymes. The results indicate that the sequential production of enzymes and cellular events leading to basidiospore formation in pileus cells are independent from each other.Abbreviation GDH_NADP NADP-dependent glutamate dehydrogenase     

Fruit body formation ofBoletus reticulatus in pure culture

The ectomycorrhyzal fungusBoletus reticulatus formed young fruit bodies in pure culture on liquid and solid media. Primordia formation started 31–32 d after inoculation on liquid medium. The primordia developed into the young fruit bodies with convex pileus and clavate stipe 44 d after inoculation on liquid medium. The ability of this fungus to form fruit bodies declined at one and half years after isolation. Sufficient nutrient in medium is required for the fungus to form mature fruit bodies in pure culture.     

Influence of light on the morphological changes that take place during the development of the <Emphasis Type="Italic">Flammulina velutipes</Emphasis> fruit body

We show that fruit bodies of Flammulina velutipes can be induced in complete darkness after a sharp temperature reduction (23° to 16°C). However, the fruit bodies that form in complete darkness have a long stipe with an undeveloped pileus on the top (pinhead fruit bodies) and are thinner and whiter than the normal fruit bodies which are formed in the light. This finding suggests that F. velutipes fruit bodies cannot mature in complete darkness. However, when we irradiated the fruit bodies that had formed in complete darkness, a pileus developed immediately, and 4 days later the separation between the stipe and the pileus could be observed. Immediately after light exposure, the stipe also thickened and became increasingly pigmented. The stipe elongation was inhibited until 8 days after light exposure, although stipe elongation progressed very quickly thereafter. Basidospores were also visible in the gills 8 days after light exposure. We consider that the basidiospore development is involved in this rapid stipe elongation, which aids the effective dispersal of basidiospores.     

The Change of Tropism in Polyporus brumalis Stipes and the Effect of Directional Stimuli on Pileus Differentiation

The directional influences of light and gravity upon the developmentof pileate and epileate sporophores of Polyporus brumalis, producedin pure culture, have been investigated. Growth in the dimiticfruit-body has a considerable subapical component which in thestipe is responsible for tropistic curvature. The stipe is competentto react negatively to unilateral gravitational and positivelyto unilateral photic stimulation throughout development, butwhen both stimulioperate phototropism masks geotropism. If illuminatedfrom one side the growing epileate stipe is strongly positivelyphototropic but as the pileus reaches a characteristic diameterthe stipe becomes negatively geotropic. Experiments with changingdirection of illumination and with artificial pilei of blackpaper suggest that the change of tropism is explicable by theshading action of the expanding pileus on the sub-pilcal photoperceptiveand photoreactive region of the stipe. Developing sporophores continuously rotated with reference tofixed directions of gravitational and light stimulation andothers with stipes inverted as a result of illumination frombelow during development have in common that the morphologicallyupper surface of the pileus always develops towards and approximatelyat right angles to the direction from which the maximum lightintensity is received. Normal but inverted stipes and dissepimentsshow no tendency to geotropic reorientation.     

Stipe elongation during basidiocarp maturation in Coprinus macrorhizus: Mechanical properties of stipe cell wall

Stipe elongation during basidiocarp maturation in the wild-type,#5026+5132, and the elongationless mutant, NG0398, of Coprinusmacrorhizus was studied, and the following results were obtained.

1. In the wild-type the middle zone of the stipe elongated 8.4times in 15 hr during maturation, while in the mutant it elongatedoaiy 2.2 times.
2. Component cells of the stipe elongated inparallel with thestipe elongation in both the wild-type andthe mutant. The widthof stipe cells was almost constant duringelongation in thewild-type, while it increased 2 times in themutant. Cell volumeincreased ca. 8 times in both stocks.
3. Theosmotic value of stipe cells was almost constant (0.45–0.50M) throughout elongation of both the wild-type and the elongationlessstipes.
4. Mechanical properties of the cell wall were examinedby measuringshrinkage, extensibility and minimum stress-relaxationtime(To) of the stipe during maturation. These parameters weredirectlyproportional to the elongation rate to follow.
5. Whenthe wild-type stipes were incubated in various concentrationsof mannitol solution and then in plain buffer solution, theextensibility of the stipe after the incubation in mannitolsolutions changed proportionally with the stipe length afterthe mannitol treatment, and To with the elongation capacityin plain buffer solution.

(Received March 3, 1977; )     

Comparative androecium morphogenesis ofSicyos angulatus andSechium edule (Cucurbitaceae)

“Androecium” ofSicyos angulatus andSechium edule is unique in having a solid central column below a head portion with thecae. Its morphogenesis was examined for the two species. The developmental course is composed of two distinct successive phases; (1) establishment of stamen primordia and (2) uplift of the stamen primordia caused by development of a central column below them. In the first phase, there is a difference between the two species; inSicyos angulatus, two bithecal and one monothecal stamen primordia are formed by congenital fusion among preformed five protrsions, whilst inSechium edule, three or four monothecal stamen primordia are formed without fusion. The central column is later produced by intercalary growth in a region below the stamen primordia in both species. Concomitant with central column development, the center of the floral primordium, which was surrounded by the early formed stamen primordia, is raised up to the top of the central column. The central column could be interpreted as a receptacular column, and not as congenitally fused stamen filaments, as currently believed. The “androecium” of the both species is considered an androecium complex, which consists of the stamens and a receptacular column.     

α1,3 Fucosyltransferase-VII modifies the susceptibility of apoptosis induced by ultraviolet and retinoic acid in human hepatocarcinoma cells

The role of α1,3fucosyltransferase-VII (α1,3 FucT-VII) in cell apoptosis was studied in human hepatocellular carcinoma H7721 cells. After the cells were transfected with α1,3 FucT-VII cDNA, the expression of apoptotic protease, procaspase-3, was decreased, while the anti-apoptotic proteins, phospho-PKB and phospho-Bad were increased as compared with mock (vector) transfected cells, indicating that α1,3FucT-VII is a potential anti-apoptotic factor in H7721 cells. After “α1,3FucT-VII” cells were irradiated by UV to induce apoptosis, the anti-apoptotic potential of α1,3FucT-VII became more apparent, as evidenced by the less apoptotic cell % and active cleaved caspase-3, more phospho-p38 MAPK and JNK (two anti-apoptotic signaling molecules in H7721 cells responsible to UV stress) when compared with the “Mock” cells. In contrast, “α1,3FucT-VII” cells facilitated the apoptosis induced by all-trans retinoic acid (ATRA), which was verified by the greater sub-G1 (apoptotic cells) peak in flow cytometry analysis, more expressions of active caspase-3 and pro-apoptotic protein Bax, as well as less expressions of anti-apoptotic proteins, Bcl-2 and Bcl-X_L. The up regulation of α1,3FucT-VII mRNA and cell surface SLe^x (α1,3FucT-VII product) by UV and down regulation of them by ATRA was speculated to be one of the mechanisms that α1,3FucT-VII decreased and increased the susceptibility of apoptosis induced by UV and ATRA respectively. Hao Wang and Qiu-Yan Wang contributed to this article equally.     

Headspace analysis identifies indole and 1-octen-3-ol as the “coal tar” odor of <Emphasis Type="Italic">Tricholoma inamoenum</Emphasis>

The odor emanating from sporocarps of Tricholoma inamoenum has been described as resembling “coal tar”. To characterize the compounds responsible for this odor, volatile chemicals released from T. inamoenum sporocarps were collected using solid phase microextraction (SPME). Subsequent analysis by gas chromatography-mass spectrometry (GC-MS) showed only indole and 1-octen-3-ol, so these compounds must be responsible for the “coal tar” odor of T. inamoenum. Mushroom pileus size was a factor in the amount of indole produced; larger mushrooms released 25-times more indole than smaller ones. A comparison of SPME and CH₂Cl₂ solvent extraction of sporocarps showed major differences in the volatile organic compounds. Benzaldehyde and phenyl acetaldehyde were the major compounds in the solvent extracts, but were not detected in the SPME experiments. Tissue disruption of the mushroom before solvent extraction showed up to a 40-fold increase in the amount of 1-octen-3-ol present.     

Four new species of <Emphasis Type="Italic">Crinipellis </Emphasis>and <Emphasis Type="Italic">Marasmius </Emphasis>in eastern Honshu,Japan

 Four new species of Crinipellis and Marasmius (Agaricales, Basidiomycetes) in eastern Honshu, Japan, are described and illustrated: (1) Crinipellis conchata sp. nov. (section Excentricinae), forming a conchate pileus and a strongly excentric, short stipe, was found on a dead twig of Trachelospermum asiaticum in Mt. Takao, Tokyo; (2) Marasmius funalis sp. nov. (section Androsacei), forming a densely white-hispid, dark brown stipe bearing numerous setiform caulocystidia, was found on a dead twig of Cryptomeria japonica or on leaf litter in Tokyo and Kanagawa; (3) Marasmius maculosus sp. nov. (section Sicci), having a relatively large, reddish-brown pileus distinctly mottled with pale colored spots and Siccus-type cheilocystidia and pileipellis cells with relatively long setulae, was found on leaf litter in the lowland forest of Kanagawa and Chiba; and (4) Marasmius sasicola sp. nov. (section Marasmius), having a small, plicate-sulcate pileus, a filiform, wiry, blackish stipe, collariate lamellae, and Siccus-type cheilocystidia and pileipellis elements, was found on fallen dead leaves of grass bamboo in Kanagawa. Received: January 30, 2002 / Accepted: May 24, 2002     

Influences of environmental factors on fruiting body induction,development and maturation in mushroom-forming fungi

Mushroom-forming fungi (restricted to basidiomycetous fungi in this review) differentiate by sensing several environmental factors for fruiting body formation. For fruiting body induction, nutrient, temperature and light conditions are critical environmental factors. Higher nitrogen and carbon sources in the media will suppress fruiting body induction in many mushroom-forming fungi, with induction being triggered by lower nitrogen and carbon concentrations. Low temperature or temperature downshift is another critical influencing factor for fruiting body induction in many cultivated mushrooms, such as Flammulina velutipes, Lentinula edodes, and Volvariella volvacea. Fungal response toward starvation and cold involves the production of sexual spores as the next generation. Species like F. velutipes and Coprinopsis cinerea can form fruiting bodies in the dark; however, light accelerates fruiting body induction in some mushroom-forming fungi. Remarkably, fruiting bodies formed in the dark have tiny or no pileus on heads (called dark stipe, pinhead fruiting body, or etiolated stipe). Light is essential for pileus differentiation in many, but not all mushroom species; one exception is Agaricus bisporus. Mushrooms have positive phototropism and negative gravitropism for effective dispersal of spores. Carbon dioxide concentrations also affect fruiting body development; pileus differentiation is suppressed at a high concentration of carbon dioxide. Thus, the pileus differentiation system of mushrooms may allow the most effective diffusion of spores. Full expansion of the pileus is followed by pileus autolysis or senescence. In C. cinerea, pileus autolysis occurs during spore diffusion. Fruiting body senescence, browning of gill, and softening occur after harvesting in several mushroom species. Fruiting body induction, development, and maturation in mushroom-forming fungi are discussed in this review.     

Distribution of free and esterified ergosterols in the medicinal fungus <Emphasis Type="Italic">Ganoderma lucidum</Emphasis>

The fruiting bodies, spores, and lipid from the spores of Ganoderma lucidum have been widely used for medicinal purpose in China. Ergosterol content may be a suitable marker for evaluating the quality of ganoderma spore and ganoderma spore lipid (GSL) products. A gradient reversed-phase high-performance liquid chromatography method was developed for the simultaneous determination of free and esterified ergosterols in G. lucidum. The contents of free and esterified ergosterols in the different parts (the stipe, pileus, tubes, and spores) of G. lucidum and GSL were determined. The results showed that total ergosterol levels in the stipe, pileus, tubes, and spores of G. lucidum were between 0.8 and 1.6 mg/g. The relative abundances of free to esterified ergosterol were different in the different parts of G. lucidum. The spores and the tubes, the hymenophore tissue that contains the spore-producing cells, have a considerably higher percentage of ergosteryl esters (41.9 and 39.7% of total ergosterol) in comparison with the pileus and stipe tissues (3.6 and 6.2%).     

Polymorphism of canonical and noncanonical <Emphasis Type="Italic">gypsy</Emphasis> sequences in different species of <Emphasis Type="Italic">Drosophila </Emphasis><Emphasis Type="Italic">melanogaster</Emphasis> subgroup: possible evolutionary relations

Mobile genetic elements constitute a substantial part of eukaryotic genome and play an important role in its organization and functioning. Co-evolution of retrotransposons and their hosts resulted in the establishment of control systems employing mechanisms of RNA interference that seem to be impossible to evade. However, “active” copies of endogenous retrovirus gypsy escape cellular control in some cases, while its evolutionary elder “inactive” variants do not. To clarify the evolutionary relationship between “active” and “inactive” gypsy we combined two approaches: the analysis of gypsy sequences, isolated from G32 Drosophila melanogaster strain and from different Drosophila species of the melanogaster subgroup, as well as the study of databases, available on the Internet. No signs of “intermediate” (between “active” and “inactive”) gypsy form were found in GenBank, and four full-size G32 gypsy copies demonstrated a convergence that presumably involves gene conversion. No “active” gypsy were revealed among PCR generated gypsy ORF3 sequences from the various Drosophila species indicating that “active” gypsy appeared in some population of D. melanogaster and then started to spread out. Analysis of sequences flanking gypsy variants in G32 revealed their predominantly heterochromatic location. Discrepancy between the structure of actual gypsy sites in G32 and corresponding sequences in database might indicate significant inter-strain heterochromatin diversity. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Voiceprint identification and its application to sociological studies of wild Japanese monkeys (Macaca fuscata yakui)

Japanese monkeys often exchange the particular vocal sound, “coo,” especially when they feed or move as a group. It was considered that the “coo” sound had no positive social meaning, perhaps because the “coo” sound network and its function were hidden behind other behavioral observations. For identification of the vocalizer only from hearing the “coo” sound, three phonetic values, i.e., the “fundamental,” “duration,” and “formants,” plus other characteristics were used as indices of voiceprints. The results indicated that these were effective for identifying the vocalizer in two-thirds of the adults in the study troop which was composed of 12 adults and 16 immature members. The “coo” sound exchange network among the troop members (adults) was drawn on the basis of the voiceprint identification. The network showed three characteristics as follows: (1) matriarchs of the kin-groups frequently exchanged “coo” sounds with each other; (2) the other females exchanged “coo” sounds mostly within their own kin-groups; and (3) males seldom participated in the “coo” sound exchange. This suggests that “coo” sound exchange plays a central role for the matriarch of kin-groups in binding each kin-group and, ultimately, in binding all members together into an organized troop.     

Differential scanning calorimetric analysis of fruit-body and mycelium ofLentinula edodes

Differential scanning calorimetric (DSC) analysis was applied to the fruit-body (pileus and stipe) and mycelium, of shiitake mushroom (Lentinula edodes). Thermograms of each sample indicated distinctive patterns. However, chemical and infrared (IR) spectroscopic analyses showed that the compositions of pileus and stipe were similar to each other and different from that of mycelium. Because the DSC thermogram depends not only on chemical composition but also on physical properties such as density, the result of DSC analysis was assumed to indicate a difference in the state of cell wall between pileus and stipe.     

Growth and fruitbody formation ofGanoderma lucidum on media supplemented with vanadium, selenium and germanium

Responses of mycelia ofGanoderma lucidum to vanadium, selenium and germanium were examined over a wide range of concentrations (10–1, 120 μg/ml) in pure culture. Se and V were found to be highly toxic, but Ge was not toxic at the levels tested.Ganododerma lucidum cultivated on substrates of sawdust with V (30–80 μg/g) developed mature fruitbodies, but the bioaccumulation of V was quite low (2.5–7 μg/g in pileus, 12.5–21.5 μg/g in stipe and <1 μg/g in basidiospores). Se as Na₂SeO₄ labeled with⁷⁵Se was effectively taken up from substrates and accumulated in fruitbodies (mainly in pileus), then depleted by discharge of basidiospores. Ge as GeCl₄ labeled with⁷⁷Ge was easily uptaken and translocated into fruitbodies.     

Three new species ofMarasmius sectionSicci from eastern Honshu, Japan

Three new species belonging toMarasmius sectionSicci (Agaricales) are described and illustrated from eastern Honshu, Japan:Marasmius nocturnus sp. nov., forming a dark brown pileus and marginate lamellae, was found on leaf litter inPasania—Quercus forests;Marasmius occultatus sp. nov., producing brownish orange or light brown basidiomata and long, cylindrical-fusoid basidiospores, was found on dead fallen twigs ofAphananthe aspera andQuercus myrsinaefolia;Marasmius opulentus sp. nov., having a reddish orange pileus and a pubescent stipe, was found on leaf litter in laurel-leaved forest.