Mature Ascocarps Research Articles

Label-free based comparative proteomic analysis of Morchella importuna development from the vegetative to the sexual reproductive stages

Morchella spp. are well-known and economically valuable edible mushrooms. The molecular mechanism of development in Morchella fungi is poorly understood, although it can be artificially cultivated. In this paper, the intracellular proteomes of three vegetative growth stages (mycelial, sclerotial initiation and maturation) and two sexual reproductive growth stages (young and mature ascocarp) of M. importuna were analyzed by LC-MS/MS. A total of 2060 credible proteins were identified. Label-free quantitative analysis showed that, consistent with physiological development, the number of differentially expressed proteins increased, which were mainly enriched in carbohydrate and amino acid metabolic pathways by GO and KAAS enrichment analysis. Besides, the categories for upregulated proteins enriched in were more in sexual reproduction stage, especially in the young ascocarp stage, than those in the vegetative growth stage, including aminoacyl-tRNA biosynthesis, histidine, peroxisome, beta-alanine metabolism, etc. In addition, the expression pattern of individual protein (top-20) suggested that the most abundant lectin protein (8.92%) played an important role in the development of M. importuna . This study comprehensively analyzed the whole developmental process of M. importuna at the proteome level, which may help us to better understand its nutrition metabolism and fruiting body development. • Intracellular proteomes of five developmental stages of M. importuna were analyzed. • Three velvet proteins were found and up-regulated in sexual reproductive stages. • A lectin protein (8.92%) was the most abundant and differentially expressed. • An abundant Woronin body major protein (3.02%) was differentially expressed.

Spatial and temporal disequilibrium of nuclear distribution in heterothallic Morchella importuna

Our previous study found that isolates from morel stipes showed a large proportion of mating type deletion, which implied the nuclear evolution and spatial heterogeneity. In this paper, the specific mating type genes of Mat1-1-1 and Mat1-2-1 in Morchella importuna were analyzed using the relative quantitative method of qRT-PCR to study the uneven nuclear distribution in morel life cycle. The results demonstrated obvious unbalanced distribution of two karyotypes in tested commercially cultivated strains and stipe issue isolates from cultivated ascocarps. The ratio of two mating type genes in tissue isolates from different parts of the ascocarps (base soil, inside and surface of stipe, inside and outside of cap pit) was also different. The disequilibrium also existed in primordium, young ascocarp and mature ascocarp. Moreover, the imbalance in different positions of mature ascocarp (base of stipe, middle of stipe, junction of stipe and cap, middle of cap, and top of cap) gradually converged from bottom to top with the largest difference in base of stipe and almost equal distribution of two mating types in morel caps, which indicated that the deviation of two kinds of nuclei in sexual reproduction stage slowed down. More than half of the protoplast regenerated strains from the heterokaryotic M04 strain were homokaryotic, with the dominant Mat1-1-1 genotype accounted for 85.7%. Overall, the Mat1-1-1 genotype nucleus dominated in all samples from different stage of the life cycle. The imbalance of nuclear distribution during the life cycle of M. importuna may imply the evolutionary-driven nuclear-level selection. This is the first study of spatial and temporal disequilibrium of nuclear distribution in macro-ascomycetes. For scale cultivation, it is necessary to perform genotype integrity testing in potential cultivated strains to ensure the existence of both mating type genes. • The two mating-type genes representing the karyotypes showed an obvious disequilibrium in the whole life cycle of Morchella importuna. • Serious disequilibrium even appear in tissue isolates, making it necessary to detect the integrity of mating type genes on isolates during spawn production. • The uneven distribution of the karyotype reflects the competition at the nuclear level.

Soil Metabarcoding Offers a New Tool for the Investigation and Hunting of Truffles in Northern Thailand.

Truffles (Tuber spp.) are well-known as edible ectomycorrhizal mushrooms, and some species are one of the most expensive foods in the world. During the fruiting process, truffles produce hypogeous ascocarps; a trained pig or dog is needed to locate the ascocarps under the ground. Truffles in northern Thailand have been recorded in association with Betula alnoides and Carpinus poilanei. In this study, we investigated the soil mycobiota diversity of soil samples from both of these truffle host plants in native forests using environmental DNA metabarcoding to target the internal transcribed spacer 1 (ITS1) region of the rDNA gene for the purposes of investigation of truffle diversity and locating truffles during the non-fruiting phase. In this study, a total of 38 soil samples were collected from different locations. Of these, truffles had been found at three of these locations. Subsequently, a total of 1341 putative taxonomic units (OTUs) were obtained. The overall fungal community was dominated by phylum-level sequences assigned to Ascomycota (57.63%), Basidiomycota (37.26%), Blastocladiomycota (0.007%), Chytridiomycota (0.21%), Glomeromycota (0.01%), Kickxellomycota (0.01%), Mortierellomycota (2.08%), Mucoromycota (0.24%), Rozellomycota (0.01%), Zoopagomycota (0.003%), and unidentified (2.54%). The results revealed that six OTUs were determined to be representative and belonged to the genus Tuber. OTU162, OTU187, OTU447, and OTU530 belonged to T. thailandicum, T. lannaense, T. bomiense, and T. magnatum, whereas OTU105 and OTU720 were acknowledged as unrecognized Tuber species. From 38 locations, OTUs of truffles were found in 33 locations (including three previously known truffle locations). Thus, 30 collection sites were considered new locations for T. thailandicum, T. bomiense, and other unrecognized Tuber species. Interestingly, at 16 new locations, mature ascocarps of truffles that were undergoing the fruiting phase were located underground. All 16 truffle samples were identified as T. thailandicum based on morphological characteristics and molecular phylogenetic analysis. However, ascocarps of other truffle species were not found at the new OTUs representative locations. The knowledge gained from this study can be used to lead researchers to a better understanding of the occurrence of truffles using soil mycobiota diversity investigation. The outcomes of this study will be particularly beneficial with respect to the search and hunt for truffles without the need for trained animals. In addition, the findings of this study will be useful for the management and conservation of truffle habitats in northern Thailand.

Different maturities drive proteomic and metabolomic changes in Chinese black truffle

Chinese black truffle (Tuber indicum) is rich in nutrition. However, commercial interests lead to the aroma components and nutrients of T. indicum being greatly affected by overexploitation without consideration of their maturity. This study investigated the proteomic and metabolomic profiles of truffle fruiting bodies at different maturities using a meta-proteomic approach. Among the 3007 identified proteins, the most up-expressed protein in the mature ascocarps was involved in the peptidyl-diphthamide biosynthetic process, while thiamine metabolism was the most differentially expressed pathway. Furthermore, a total of 54 metabolites identified upon LC-MS differed significantly, with 30 being up-expressed in the mature ascocarps, including organic acids, carnitine substances and polysaccharides. Additionally, the ash, protein, fat, crude fiber and total sugar contents were all higher in the mature ascocarps. Overall, our findings reveal that mature truffles have a higher nutritional value, providing a basis for further exploring protein functionality of T. indicum at different maturities.

Controlled surface fire for improving yields of Morchella importuna

Fire is known to benefit morel production, but the details of how or why remain unknown. To investigate the effects of fire treatment on the occurrence and yield of the morel Morchella importuna in semi-artificial, field-based setups, we designed surface fire treatments using different quantities of burning material (dry Pinaceae twigs). Developmental stages of M. importuna, including the formation and degeneration of mycelium, formation of ascocarp primordia, ascocarp initials, and mature ascocarps, were evaluated in square-meter quadrats in field experiments. Morel yields and soil were compared. Fire treatments corresponding to 1.7 and 2.5 kg burning material/m2 resulted in clearly increased levels of soil minerals and nutrients, and significantly increased formation of ascocarp primordial and morel yields. None of the fire treatments had a significant effect on the germination rate of primordia. Fire treatments corresponding to 0.8 kg burning material/m2 did not increase morel yield. Surface fires increased the suitability of soils for the growth and productivity of ascocarps of M. importuna if sufficient burning material is used.

Light and scanning electron microscopy studies of the early infection stages of Hymenoscyphus pseudoalbidus on Fraxinus excelsior

The ontogeny and morphology of infection structures associated with the early stages of infection of Hymenoscyphus pseudoalbidus on common ash (Fraxinus excelsior) leaves and leaf petioles were investigated using light and scanning electron microscopy. Ascospores were produced in mature ascocarps and infections on ash leaves were first observed 2 weeks later. Ascospores developed germ tubes, followed by appressorium formation and penetration of epidermal cells on ash leaves and petioles. Chalara fraxinea spores, the anamorph of H. pseudoalbidus, appeared and were arranged in chains, surrounded by a membranous sheath, and varied considerably in size and shape. Host invasion and colonization of all cell types of leaves and petioles were observed using light microscopy. The role of leaves and petioles as sites of infection in the life cycle of H. pseudoalbidus and the disease cycle of ash dieback is discussed.

Differential Gene Expression During Conidiation in the Grape Powdery Mildew Pathogen, Erysiphe necator

Asexual sporulation (conidiation) is coordinately regulated in the grape powdery mildew pathogen Erysiphe necator but nothing is known about its genetic regulation. We hypothesized that genes required for conidiation in other fungi would be upregulated at conidiophore initiation or full conidiation (relative to preconidiation vegetative growth and development of mature ascocarps), and that the obligate biotrophic lifestyle of E. necator would necessitate some novel gene regulation. cDNA amplified fragment length polymorphism analysis with 45 selective primer combinations produced ≈1,600 transcript-derived fragments (TDFs), of which 620 (39%) showed differential expression. TDF sequences were annotated using BLAST analysis of GenBank and of a reference transcriptome for E. necator developed by 454-FLX pyrosequencing of a normalized cDNA library. One-fourth of the differentially expressed, annotated sequences had similarity to fungal genes of unknown function. The remaining genes had annotated function in metabolism, signaling, transcription, transport, and protein fate. As expected, a portion of orthologs known in other fungi to be involved in developmental regulation was upregulated immediately prior to or during conidiation; particularly noteworthy were several genes associated with the light-dependent VeA regulatory system, G-protein signaling (Pth11 and a kelch repeat), and nuclear transport (importin-β and Ran). This work represents the first investigation into differential gene expression during morphogenesis in E. necator and identifies candidate genes and hypotheses for characterization in powdery mildews. Our results indicate that, although control of conidiation in powdery mildews may share some basic elements with established systems, there are significant points of divergence as well, perhaps due, in part, to the obligate biotrophic lifestyle of powdery mildews.

Grapevine powdery mildew (Erysiphe necator): a fascinating system for the study of the biology, ecology and epidemiology of an obligate biotroph.

Few plant pathogens have had a more profound effect on the evolution of disease management than Erysiphe necator, which causes grapevine powdery mildew. When the pathogen first spread from North America to England in 1845, and onwards to France in 1847, 'germ theory' was neither understood among the general populace nor even generally accepted within the scientific community. Louis Pasteur had only recently reported the microbial nature of fermentation, and it would be another 30 years before Robert Koch would publish his proofs of the microbial nature of certain animal diseases. However, within 6 years after the arrival of the pathogen, nearly 6 million grape growers in France were routinely applying sulphur to suppress powdery mildew on nearly 2.5 million hectares of vineyards (Campbell, 2006). The pathogen has remained a focus for disease management efforts ever since. Because of the worldwide importance of the crop and its susceptibility to the disease, and because conventional management with modern, organic fungicides has been compromised on several occasions since 1980 by the evolution of fungicide resistance, there has also been a renewed effort worldwide to explore the pathogen's biology and ecology, its genetics and molecular interactions with host plants, and to refine current and suggest new management strategies. These latter aspects are the subject of our review. The most widely accepted classification follows. Family Erysiphaceae, Erysiphe necator Schw. [syn. Uncinula necator (Schw.) Burr., E. tuckeri Berk., U. americana Howe and U. spiralis Berk. & Curt; anamorph Oidium tuckeri Berk.]. Erysiphe necator var. ampelopsidis was found on Parthenocissus spp. in North America according to Braun (1987), although later studies revealed isolates whose host range spanned genera, making the application of this taxon somewhat imprecise (Gadoury and Pearson, 1991). The classification of the genera before 1980 was based on features of the mature ascocarp: (i) numbers of asci; and (ii) morphology of the appendages, in particular the appendage tips. The foregoing has been supplanted by phylogeny inferred from the internal transcribed spacer (ITS) of ribosomal DNA sequences (Saenz and Taylor, 1999), which correlates with conidial ontogeny and morphology (Braun et al., 2002). The pathogen is obligately parasitic on genera within the Vitaceae, including Vitis, Cissus, Parthenocissus and Ampelopsis (Pearson and Gadoury, 1992). The most economically important host is grapevine (Vitis), particularly the European grape, V. vinifera, which is highly susceptible to powdery mildew. Disease symptoms and signs: In the strictest sense, macroscopically visible mildew colonies are signs of the pathogen rather than symptoms resulting from its infection, but, for convenience, we describe the symptoms and signs together as the collective appearance of colonized host tissues. All green tissues of the host may be infected. Ascospore colonies are most commonly found on the lower surface of the first-formed leaves near the bark of the vine, and may be accompanied by a similarly shaped chlorotic spot on the upper surface. Young colonies appear whitish and those that have not yet sporulated show a metallic sheen. They are roughly circular, ranging in size from a few millimetres to a centimetre or more in diameter, and can occur singly or in groups that coalesce to cover much of the leaf. Senescent colonies are greyish, and may bear cleistothecia in various stages of development. Dead epidermal cells often subtend the colonized area, as natural mortality in the mildew colony, the use of fungicides, mycoparasites or resistance responses in the leaf result in the deaths of segments of the mildew colony and infected epidermal cells. Severely affected leaves usually senesce, develop necrotic blotches and fall prematurely. Infection of stems initially produces symptoms similar to those on leaves, but colonies on shoots are eventually killed as periderm forms, producing a dark, web-like scar on the cane (Gadoury et al., 2011). Inflorescences and berries are most susceptible when young, and can become completely coated with whitish mildew. The growth of the berry epidermal tissue stops when severely infected, which may result in splitting as young fruit expand. Berries in a transitional stage between susceptible and resistant (generally between 3 and 4 weeks after anthesis) develop diffuse, nonsporulating mildew colonies only visible under magnification. Diffuse colonies die as berries continue to mature, leaving behind a network of necrotic epidermal cells (Gadoury et al., 2007). Survival over winter as mycelium in buds results in a distinctive foliar symptom. Shoots arising from these buds may be heavily coated with fungal growth, stark white in colour and stand out like white flags in the vine, resulting in the term 'flag shoots'. More commonly, colonization of a flag shoot is less extensive, and infection of a single leaf, or of leaves on one side of the shoot only, is observed (Gadoury et al., 2011).

Sexual reproduction in aflatoxin-producing Aspergillus nomius

Sexual reproduction was examined in the aflatoxin-producing fungus Aspergillus nomius. Crosses between sexually compatible strains resulted in the formation of multiple nonostiolate ascocarps within stromata, which places the teleomorph in genus Petromyces. Ascocarp and ascospore morphology in Petromyces nomius were similar to that in P. flavus and P. parasiticus, and differences between teleomorphs were insufficient for species separation. Formation of mature ascocarps was infrequent, with only 24% of the 83 crosses producing viable ascospores. The majority of P. nomius strains contained a single mating-type gene (MAT1-1 or MAT1-2), but several strains contained both genes. MAT1-1/MAT1-2 strains were self-sterile and capable of mating with both MAT1-1 and MAT1-2 strains; hence P. nomius appears to be functionally heterothallic.

Initiation, Development, and Survival of Cleistothecia of Podosphaera aphanis and Their Role in the Epidemiology of Strawberry Powdery Mildew

A collection of four clonal isolates of Podosphaera aphanis was heterothallic and was composed of two mutually exclusive mating types. Cleistothecial initials approximately 20 to 30 microm in diameter were observed within 7 to 14 days after pairing of compatible isolates and developed into morphologically mature ascocarps within 4 weeks after initiation on both potted plants maintained in isolation and in field plantings in New York State and southern Norway. Ascospores progressed through a lengthy maturation process over winter, during which (i) the conspicuous epiplasm of the ascus was absorbed; (ii) the osmotic potential of the ascospore cytoplasm increased, resulting in bursting of prematurely freed spores in water; and, finally, (iii) resulting in the development of physiologically mature, germinable, and infectious ascospores. Release of overwintered ascospores from field collections was coincident with renewed plant growth in spring. Overwintered cleistothecia readily dehisced when wetted and released ascospores onto glass slides, detached strawberry leaves, and leaves of potted plants. Plant material exposed to discharged ascospores developed macroscopically visible mildew colonies within 7 to 10 days while noninoculated controls remained mildew free. Scanning electron and light microscopy revealed that cleistothecia of P. aphanis were enmeshed within a dense mat of hyphae on the persistent leaves of field-grown strawberry plants and were highly resistant to removal by rain while these leaves remained alive. In contrast, morphologically mature cleistothecia on leaves of nine deciduous perennial plant species were readily detached by simulated rain and seemed adapted for passive dispersal by rain to other substrates. Contrary to many previous reports, cleistothecia appear to be a functional source of primary inoculum for strawberry powdery mildew. Furthermore, they differ substantially from cleistothecia of powdery mildews of many deciduous perennial plants in their propensity to remain attached to the persistent leaves of their host during the intercrop period.

Distribution of mating types and genetic diversity induced by sexual recombination in Setosphaeria turcica in northern China

Mature ascocarps and ascospores in the heterothallic ascomycete fungus, Setosphaeria turcica, were successfully produced in Sach’s medium with barley culm as the mating stimulator after four weeks’ coincubation of two opposite mating type isolates at 25°C in darkness. A single isolate could not produce ascospores or ascocarps. The ascocarps were produced on the exposed surface and embedded parts of barley culm or in the upper layer of the medium. The asci linked themselves to ascocarp with their short handles and assembled at the bottom of the ascocarp. Many asci had four to six colorless mature ascospores with one to six septa. But asci with eight ascospores were also found. Using isolate 9914 and isolate 9961 as standard testers for mating types (MAT1 and MAT2), respectively, 94 isolates of S. turcica collected from northern China in 1999, 2003, and 2004 were grouped into three mating types: MAT1 (53 isolates), MAT2 (31 isolates) and MAT12 (10 isolates). The MAT12 isolates, which were first found in China, were compatible with not only MAT1 isolates but also MAT2 isolates. No MAT12 isolates were found in 1999, but 2 MAT12 isolates and 8 MAT12 isolates were found in 2001 and 2003, respectively. The geographic distribution of different mating types was unequal among locations. Generally the frequency of MAT1 was significantly higher than that of MAT2 and MAT12. The unequal distribution of mating types suggested a low frequency of genetic recombination. The pathogenicity of different mating type isolates was tested on the susceptible corn inbred B37 and the results revealed that the disease latency period, disease incidence, lesion area and conidia production were not significantly different among the three mating type groups. However, the pathogenicity of the progeny isolates of isolate 99-12 (MAT2, race 1) and isolate 99-15 (MAT1, race 0) was significantly different from the parent isolates, isolate 99-12 and isolate 99-15, suggesting that sexual recombination could cause significantly virulence variation in S. turcica. Random amplification of polymorphic DNA (RAPD) analysis also revealed high genotype diversity among the progeny isolates, indicating that the sexual recombination could also produce significant genetic variation in the fungal pathogen.

Occurrence and diversity of bacterial communities in Tuber magnatum during truffle maturation

Tuber magnatum, an ascomycetous fungus and obligate ectomycorrhizal symbiont, forms hypogeous fruit bodies, commonly called Italian white truffles. The diversity of bacterial communities associated with T. magnatum truffles was investigated using culture-independent and -dependent 16S rRNA gene-based approaches. Eighteen truffles were classified in three groups, representing different degrees of ascocarp maturation, based on the percentage of asci containing mature spores. The culturable bacterial fraction was (4.17 +/- 1.61) x 10(7), (2.60 +/- 1.22) x 10(7) and (1.86 +/- 1.32) x 10(6) cfu g(-1) for immature, intermediate and mature ascocarps respectively. The total of bacteria count was two orders of magnitude higher than the cfu g(-1) count. Sequencing results from the clone library showed a significant presence of alpha-Proteobacteria (634 of the 771 total clones screened, c. 82%) affiliated with Sinorhizobium, Rhizobium and Bradyrhizobium spp. The bacterial culturable fraction was generally represented by gamma-Proteobacteria (210 of the 384 total strains isolated, c. 55%), which were mostly fluorescent pseudomonads. Fluorescent in situ hybridization confirmed that alpha-Proteobacteria (85.8%) were the predominant components of truffle bacterial communities with beta-Proteobacteria (1.5%), gamma-Proteobacteria (1.9%), Bacteroidetes (2.1%), Firmicutes (2.4%) and Actinobacteria (3%) only poorly represented. Molecular approaches made it possible to identify alpha-Proteobacteria as major constituents of a bacterial component associated with T. magnatum ascoma, independently from the degree of maturation.

The ontogeny of perithecia in Guignardia bidwellii

The ontogeny of perithecia in Guignardia bidwellii was studied with contaminated berries and under in vitro conditions. The first developmental stage consists of a stromatic envelope surrounding a carpocentrum in which an ascogonial apparatus (composed of ascogonial cells and trichogynes) differentiates at the second stage. Stage 3 is marked by the incipient formation of the ascal locule and development of pseudoparaphyses. At the subadult stage ( = stage 4) these filaments intermingle with asci and they are progressively stretched owing to ascus expansion. The mature ascocarp (stage 5) contains numerous eight-spored asci and opens through an apical ostiole. The developmental features of Guignardia bidwellii indicate a pseudoparaphysate ascolocular type of development. Consequently, the fungus is more correctly placed in the Botryosphaeriaceae than in the Mycosphaerellaceae.

THE CONTROL OF SEXUAL MORPHOGENESIS IN THE ASCOMYCOTINA

Summary(1) A series of factors controls sexual morphogenesis in the Ascomycotina, a process involving the formation of novel structures such as ascocarps (fruit bodies) and asci (sacs containing spores) during sexual reproduction.(2) Environmental and genetic factors must be correct before Ascomycetes may sexually reproduce. Compatibility in many heterothallic species is under polygenic control, with the mating type loci and also other genetic factors determining the productivity of sexual crosses.(3) Classical genetic studies have shown that sexual morphogenesis involves the expression of a series of developmentally regulated genes, and this has been confirmed by recent molecular studies which have demonstrated changes in patterns of mRNA and protein synthesis during ascocarp formation.(4) Hyphal differentiation leading to the formation of mature fruit bodies occurs in response to a series of signals, which include various physical and chemical factors.(5) Chemical sex factors have been identified which are believed to have important regulatory or nutritional roles in sexual morphogenesis. These include the following.(a) Diffusible sex hormones which may regulate developmental switching between asexual and sexual modes of reproduction, including (i) pheromones involved with the induction of gametangia and gamete attraction, and (ii) sex morphogens involved with triggering particular stages of fruit body formation. (b) Sexual growth substances which are required as nutrients, and may be precursors for the production of sex hormones, or metabolites used in the synthesis of novel sexual structures. Most of these sex factors are lipids.(6) Certain sex morphogens and sexual growth substances have been shown to exhibit activity in a variety of fungal species, suggesting that fungi of related phylogenetic descent may utilize similar metabolites or signalling factors during sexual reproduction.(7) Phenoloxidase enzymes may catalyse hyphal aggregation in developing fruit bodies.(8) Initial stages of ascocarp development may occur independently of the events of the sexual cycle. However, a link(s) with the functional ascogenous hyphae is needed for the formation of morphologically mature ascocarps.(9) Suitable environmental conditions are sufficient to trigger sexual morphogenesis in homothallic Ascomycetes. However, an extra level of control is present in heterothallic species, with a compatible partner required to complete sexual reproduction. This may be partly because novel regulatory products, formed by the combined action of the mating type loci of different partners, are required for further ascocarp development.(10) Further research is required to identify more fungal chemical sex factors and to determine the role of environmental stress in controlling sexual morphogenesis, and how this may be related to temporal patterns in the expression of mating type genes.

Ethylene production by ascocarps of Elaphomyces species.

Ascocarps of the hypogaean fungus Elaphomyces spp. from some Swedish forest stands were examined for production of ethylene. Significant amounts were produced by mature ascocarps of E. asperulus and E. granulatus. E. muricatus, which was found only in juvenile or immature stages, did not produce ethylene. None of the ascocarps studied were infected by Cordyceps. The ecological role of the ethylene production by the ascocarps is shortly discussed.

Ascocarp production of Magnaporthe salvinii in culture

Nutritional and physical factors affecting ascocarp production of the causal agent of rice stem rot, Magnaporthe salvinii , were studied in a cross between Japanese isolates. Mature ascocarps were produced on several agar media with rice culms. The optimum temperatures for mature ascocarp production were from 22 to 26 °C. Light and dark greatly influenced ascocarp production, since no mature ascocarps were produced in continuous darkness.

Factors influencing ascocarp formation in Polystigma ochraceum

The effects of leaf age, soil depth, and incubation temperature on ascocarp production by Polystigma ochraceum (Wahl.) Sacc, the causal agent of almond leaf blight were studied under field and laboratory conditions. Mature ascocarps were formed in leaves harvested from July to October (leaf fall) but more were found in October samples. Ascocarps were produced under field conditions in infected leaves buried in soil at the depths of o to 40 cm. with maximum mature ascocarp formation at 5 cm and minimum at the soil surface. No ascocarp was observed in leaves incubated at temperatures above 10° C. At 5°, the number of mature ascocarps increased as the incubation period was extended.

New or Interesting British Microfungi, III

tearing open irregularly, surface smooth, rather soft when fresh, scarlet-red (Ridgway), developing apparently at or just below soil level, sometimes attached to plant debris but not to living roots. Mature ascocarps, including all the English material seen, the type, and most New Zealand collections at Kew, consist of a pseudoparenchymatic rind about 300 [ thick enclosing a large cavity lined with cream-coloured powder consisting of loose spores entangled amongst slender hyaline hyphae. In this state no asci can be found though, as Patouillard showed, spores can often be seen to cohere in rows. A single New Zealand collection, Waitahene Road, Auckland, April 1962, contains an immature ascocarp, about 15 mm diameter in the dried state, with smooth unwrinkled surface and thin red rind enclosing a solid white flesh permeated by rather sparse yellowish veins. The detailed structure of the gleba is difficult to elucidate from dried material and needs full investigation on living specimens. Innumerable, more or less cylindrical, long-stalked asci are present, however, often with an irregular or undulating outline but apparently lying more or less parallel in the matrix. Asci of very different sizes and states of development occur together. Those teased out measure about 200-300 X 20-25 u, with flat tips and non-amyloid walls. Mature asci often contain only four ascospores but up to six well-developed ascospores can be found and sometimes spores of very different sizes, as though some were degenerating. The asci are surrounded by slender, branched,

Neoxenophila foetida gen. et sp.nov., a new keratinophilic ascomycete

A new keratinophilic ascomycete, Neoxenophila foetida gen. et sp.nov., conidial state Chrysosporium foetidum Apinis & Clark, is described. It was isolated from a rat, Myomyces daltoni , in Nigeria. Mature ascocarps of the fungus are produced on certain laboratory media after mating two sexually different isolates, D 3c and D 3g . The fungus described is related to Aphanoascus Zukal and Xenophila Malloch.

A comparison of saline tolerance and sporulation in marine and clinical isolates of Allescheria boydii shear

Two marine and 14 clinical isolates ofA. boydii and its byssoid phase,M. apiospermum, were compared with respect to their saline tolerance, morphological development, and sporulation on various commercial and compounded media. Isolates from sea-water and human infections were morphologically alike, and possessed similar saline tolerance. Most strains had salinity optima near 9‰, about the salinity of blood. Only marine isolates fruited well on Sabouraud's dextrose agar and other routine mycological media. On a medium containing wood and sea-water, and few soluble organics, 12 strains formed mature ascocarps or synnemata, or both. Low nutrient level media containing wood or cellulosic paper, with or without sea-water, were recommended for the propagation and conservation of sporulating cultures of many saprobic Ascomycetes and synnematous Fungi Imperfecti.