Relations entre production et localisation de mycosporine et morphogenèses reproductrices chez le Pyrénomycète Gnomonia leptostyla

Relations between production and localisation of mycosporin and reproductive morphogenesis in the Pyrenomycete Gnomonia leptostyla.
The production of mycosporin (P310) has been analysed in Gnomonia leptostyla (FT.) Ces. et de Not. during mycelial growth and reproductive morphogenesis (macroconidiogenesis, microconidiogenesis and differentiation of perithecia). Conidiogenesis is induced in illuminated cultures while darkness promotes perithecial development. At 20°C, the cultures produce either macroconidia or perithecia with abortive sporophyte. Microconidia differentiation and perithecia maturation require low temperature (10°C). Mycosporin is, at all times, present in the thallus. However, the concentration of mycosporin in highest in the conidiogenous thallus, intermediate in the perithecial thallus. and lowest in the vegetative mycelium. In the conidiogenous thallus, macroconidia and microconidia are both sites of mycosporin accumulation. On the contrary, in the perithecial thallus, mycosporin levels are not higher in perithecia than in mycelia, even during their maturation period. The quantitative variations of mycosporin during the thallus development and its accumulation inside conidia suggest translocation from sites of synthesis towards reproductive cells.     

Phenotypic Diversity among Alleles at the PER-1 Locus of NEUROSPORA CRASSA

Comparison of 11 perithecial color mutants suggested that all were alleles at the per-1 locus but nonetheless separable into two groups because of phenotypic differences. Three of the mutant strains produced orange perithecia and black ascospores, and eight produced paler, yellow perithecia and white ascospores. Perithecial phenotype was dependent upon the genotype of the protoperithecial parent; ascospore phenotype, upon the genotype of the individual ascospore. No evidence was found that the white ascospores were due to chromosomal rearrangements. No separation of the perithecial and ascospore phenotypes by recombination was observed in a cross between one of the mutants and a per-1+ strain. However, apparent low levels of recombination in crosses between some of the mutants indicated possible genetic complexity at the per-1 locus. The phase specificity of the per-1 mutations and the possible nature and mode of expression of the orange and yellow perithecial pigments are discussed.     

Scanning electron microscopy observations of the interaction between Trichoderma harzianum and perithecia of Gibberella zeae

Chronological events associated with the interaction between a strain of Trichoderma harzianum, T472, with known biological control activity against perithecial production of G. zeae, were studied with scanning electron microscopy to investigate the mechanisms of control. Large clusters of perithecia consisting of 5-15 perithecia formed on the autoclaved, mulched wheat straw inoculated with G. zeae alone (control) with an average of 157 perithecia per plate. Small clusters consisting of 3-6 and an average of 15 perithecia per plate perithecia formed on straw that was treated with T. harzianum. The mature perithecia from straw treated with T. harzianum produced less pigment and were lighter in color than those from the control plates. Furthermore the cells of the outer wall of these perithecia were abnormal in appearance and unevenly distributed across the surface. Immature perithecia were colonized by T. harzianum approximately 15 d after inoculation (dai) with the biocontrol agent and pathogen. Few perithecia were colonized at later stages. The affected perithecia collapsed 21 dai, compared to the perithecia in the control samples that began to collapse 28 dai. Abundant mycelium of T. harzianum was seen on the perithecia of treated samples. Perithecial structures may be resistant to penetration by the mycelium because direct penetration was not observed. Trichoderma harzianum colonized the substrate quickly and out-competed the pathogen, G. zeae.     

Scanning electron microscopy of surface and internal features of developing perithecia of Neurospora crassa.

Stages in the development of perithecia of Neurospora crassa, designated by the time elapsed after crossing, were investigated with the scanning electron microscope, from protoperithecia through perithecia. The usual examination of external features of whole specimens with this instrument was augmented by a freeze-fracture technique which allowed the viewing of development internally as well. Rapid increases in perithecial size soon after crossing were followed by the appearance, in section, of a centrum, at first undifferentiated but subsequently developing ascogenous hyphae. The perithecial beak appeared as a compact mass easily distinguishable in whole specimens from the surrounding hyphae by means of texture as well as shape. Two ascospores were photographed during emergence from an ostiole, but ostioles were found more frequently closed than open.     

Stemphylium leaf blight of onion

A foliar blight disease of onion (Allium cepa L.) and garlic (Allium sativum L.) hitherto unreported from India has been described from Varanasi, U. P. The infection is confined to the leaves and inflorescence stalks. The conidial stage of the pathogen is more injurious to the plants, whereas the perithecial stage occurs on the inflorescence stalks in isolated locations. The conidiophores and conidia typical of the genus Stemphylium Wallroth are formed during the early development of the disease followed by perithecia predominantly developing on the peduncles. The perithecia were also induced to develop in artificial culture and were identical in diagnostic characters to those from the field. The conidia and ascospores from both host and artificial culture were reciprocally pathogenic to onion and garlic. Morphology of the fruiting structures indicated identity of the pathogen in its conidial stage with Stemphylium vesicarium (Wallroth) Simmons and in the perithecial stage with Pleospora allii (Rabenh.) Ces. and de Not., to which they are referred respectively.     

Experiments using sterilized apple-leaf discs to study the mode of action of urea in suppressing perithecia of Venturia inaequalis (Cke.) Wint.

Laboratory experiments using sterilized apple-leaf discs showed that treatment of leaves with urea during the early stages of perithecial initiation induced a high nitrogen content of the leaves, which prevented further development of perithecia although mycelial growth was unaffected. Treatments applied at a later stage of fungal development inhibited both perithecial and mycelial growth. Some of the bacteria isolated from urea-treated leaves in the field restricted perithecial development, particularly when applied in the first month after inoculation with suspensions of conidia from sexually compatible strains of the fungus. One isolate, a Pseudomonas sp., was shown to be important in the decomposition of apple leaves.     

Scanning Electron Microscopy on the Perithecial Development of Phyllactinia corylea on Mulberry-II. Sexual Stage

Development of perithecia of Phyllactinia corylea (Pers.) Karst. on mulberry (Morus spp.) leaves was examined by scanning electron microscopy. Two short specialized structures, antheridium and ascogonium, emerged from two separate hyphae, were fused with each other and developed into an egg‐shaped perithecial primordia. These primordia later developed into globose immature perithecia, which covered with protruded wall cells with clear margins. A large number of hyphae emerged from near the base of globose perithecia, which radiated on the leaf surface and thus helped the perithecia to fix to the surface. Specific characteristic penicillate cells and acicular appendages originated from the immature perithecia. The penicillate cells developed with apical sterigma‐like projections from the wall cells of the upper part of immature perithecia, whereas the acicular appendages originated from the shrunken wall cells at the perithecial equatorial planes. On maturation of perithecia, the acicular appendages bent down and pushed the perithecia above the substrate and thus helped them to liberate out. The sterigma‐like projections were covered with paste‐like granular substance, which help the dispersed perithecia to attach to mulberry leaves and branches.     

Effect of Trichoderma harzianum on perithecial production of Gibberella zeae on wheat straw

Conidial suspensions and cell-free filtrates of Trichoderma harzianum isolates were evaluated for their effectiveness in reducing perithecial and ascospore production of Gibberella zeae on wheat straw. Isolate T-22, which is registered in the US as a biological control agent (Plant Shield™), was included in the study as a positive control. When co-inoculated with G. zeae all 11 isolates of T. harzianum significantly reduced perithecial numbers on wheat straw. Five T. harzianum isolates, including T-22, reduced perithecial formation by 70% or greater. All isolates of G. zeae, varied in their ability to produce perithecia. Isolate 192132 produced the greatest number of perithecia and was used to further evaluate the effect of application time of the T. harzianum isolates. Perithecial reduction was highest (96-99%) when T. harzianum conidial suspension or cell-free filtrate was applied to straw 24 h prior to inoculation with G. zeae. Control was less effective when T. harzianum was applied at the same time (co-inoculated) or 24 h after G. zeae. Treatments which reduced perithecial numbers also reduced ascospore numbers; however, the average numbers of ascospores per perithecia were not significantly lowered. Field trials showed significant reduction of perithecia on residues treated with T. harzianum prior to placement on the soil surface. Both T. harzianum and G. zeae were re-isolated from residues sampled in July and August after 30 and 60 days of exposure to the environment.     

Roles for receptors, pheromones, G proteins, and mating type genes during sexual reproduction in Neurospora crassa

Here we characterize the relationship between the PRE-2 pheromone receptor and its ligand, CCG-4, and the general requirements for receptors, pheromones, G proteins, and mating type genes during fusion of opposite mating-type cells and sexual sporulation in the multicellular fungus Neurospora crassa. PRE-2 is highly expressed in mat a cells and is localized in male and female reproductive structures. Δpre-2 mat a females do not respond chemotropically to mat A males (conidia) or form mature fruiting bodies (perithecia) or meiotic progeny (ascospores). Strains with swapped identity due to heterologous expression of pre-2 or ccg-4 behave normally in crosses with opposite mating-type strains. Coexpression of pre-2 and ccg-4 in the mat A background leads to self-attraction and development of barren perithecia without ascospores. Further perithecial development is achieved by inactivation of Sad-1, a gene required for meiotic gene silencing. Findings from studies involving forced heterokaryons of opposite mating-type strains show that presence of one receptor and its compatible pheromone is necessary and sufficient for perithecial development and ascospore production. Taken together, the results demonstrate that although receptors and pheromones control sexual identity, the mating-type genes (mat A and mat a) must be in two different nuclei to allow meiosis and sexual sporulation to occur.     

New species from the Fusarium solani species complex derived from perithecia and soil in the old World tropics

A large collection of strains belonging to the Fusarium solani species complex (FSSC) was isolated from soil and perithecia in primary forests in Sri Lanka (from fallen tree bark) and tropical Australia (Queensland, from fallen tree fruits and nuts). Portions of the translation elongation factor 1-alpha (tef1) gene, the nuclear large subunit (NLSU) and internal transcribed spacer regions (ITS) of the nuclear ribosomal RNA genes were sequenced in 52 isolates from soil and perithecia. The FSSC was divided previously into three clades with some biogeographic structure, termed Clades 1, 2 and 3. All Sri Lankan and Australian soil isolates were found to be members of Clade 3, most grouping with the cosmopolitan soil-associated species F. falciforme. All but two Sri Lankan perithecial isolates were associated with a set of five divergent phylogenetic lineages that were associated with Clade 2. Australian perithecial isolates resided in a subclade of Clade 3 where most of the previously defined mating populations of the FSSC reside. Isolates from perithecia and those cultured from soil were always members of different species lineages, even when derived from proximal locations. The previous biogeographic assignment of Clade 2 to South America is now expanded to the worldwide tropics. Sri Lanka appears to be an important center of diversity for the FSSC. Nectria haematococca is epitypified with a collection from the type locality in Sri Lanka; its anamorph is described as a new species, Fusarium haematococcum. Neocosmospora E.F. Smith is adopted as the correct genus for Nectria haematococca. These new species are described: F. kurunegalense/Neo. kurunegalensis, F. rectiphorus/Neo. rectiphora/, F. mahasenii/Neo. mahasenii/, F. kelerajum/Neo. keleraja.     

PERITHECIAL FORMATION IN GELASINOSPORA RETICULISPORA. I. EFFECTS OF LIGHT AT TWO DIFFERENT GROWTH STATES

Hyphae of Gelasinospora reticulispora were cultured on corn meal agar in a growth tube at 25 ± 0.4°C under different light conditions. While the hyphal tip was growing, perithecia were not formed under continuous white light (ca. 2000 ergs cm^?2 sec^?1), but some perithecia were initiated in total darkness. However, when white light was given after a dark period, perithecial formation was greatly promoted. In these cases, perithecial formation occurred in older portion of the culture (the portion nearest the point of inoculation) at first, and then gradually spread to the younger portion. Immediately after the tip of hyphae reached the other end of the growth tube, perithecia were induced in the youngest portion of the hyphae irrespective of the photoconditions; then formation proceeded toward the older portion. This induction was not age-dependent, because in growth tubes with different lengths, perithecia always became visible ca. 24 hr after the tip of hyphae reached the other end of growth tube. The photoinhibitory effect was no longer observed thereafter, but photopromotive effect was still evident.     

Perithecial formation in Gelasinospora reticulispora III. Inhibitory effects of near-UV and blue light during the inductive dark period

Growing hyphae of Gelasinospora reticulispora required a continuousdark period prior to photoinduction of perithecia. The inductivedark period was interrupted by brief exposure of the hyphaeto white light so that the formation of perithecia no longertook place. Photosensitivity of the hyphae in terms of the light-breakeffect gradually changed during the inductive dark period. Sensitivityreached its maximum at the 18th hr of the dark period when anirradiation of 1?10⁵ ergs cm^–2 of near-UV light or 4?10⁴ergs cm^–2 of blue-light was sufficient for the light-break.Red and far-red light had no effect at all. The light-breakeffect was limited to the irradiated portion of the hyphae anddid not affect any unirradiated portions. Inhibitory effecton perithecial formation of continuous white light could betotally replaced for several days with intermittent irradiationof near-UV or blue light if given for 5 min every 4 hr. (Received December 18, 1973; )     

Autophagy provides nutrients for nonassimilating fungal structures and is necessary for plant colonization but not for infection in the necrotrophic plant pathogen Fusarium graminearum

The role of autophagy in necrotrophic fungal physiology and infection biology is poorly understood. We have studied autophagy in the necrotrophic plant pathogen Fusarium graminearum in relation to development of nonassimilating structures and infection. We identified an ATG8 homolog F. graminearum ATG8 whose first 116 amino acids before the predicted ATG4 cleavage site are 100% identical to Podospora anserina ATG8. We generated a ΔFgatg8 mutant by gene replacement and showed that this cannot form autophagic compartments. The strain forms no perithecia, has reduced conidia production and the aerial mycelium collapses after a few days in culture. The collapsing aerial mycelium contains lipid droplets indicative of nitrogen starvation and/or an inability to use storage lipids. The capacity to use carbon/energy stored in lipid droplets after a shift from carbon rich conditions to carbon starvation is severely inhibited in the ΔFgatg8 strain demonstrating autophagy-dependent lipid utilization, lipophagy, in fungi. Radial growth rate of the ΔFgatg8 strain is reduced compared with the wild type and the mutant does not grow over inert plastic surfaces in contrast to the wild type. The ability to infect barley and wheat is normal but the mutant is unable to spread from spikelet to spikelet in wheat. Complementation by inserting the F. graminearum atg8 gene into a region adjacent to the actin gene in ΔFgatg8 fully restores the WT phenotype. The results showed that autophagy plays a pivotal role for supplying nutrients to nonassimilating structures necessary for growth and is important for plant colonization. This also indicates that autophagy is a central mechanism for fungal adaptation to nonoptimal C/N ratios.     

Environmental factors affecting the degradation of Dyfonate by soil fungi.

The ability of selected fungi to degrade the soil insecticide Dyfonate (O-ethyl S-phenyl ethylphosphonodithioate) into water-soluble, noninsecticidal metabolites was found to be dependent on the supply of nutrients, incubation time, temperature, pH, as well as other factors. With yeast extract as the carbon source (5 g/liter) and ammonium nitrate (1 g/liter) as the nitrogen source, both Rhizopus arrhizus and Penicillium notatum degraded the insecticide to a larger extent than with any other combination of nutrients used. With glucose as the carbon source, concentrations of ammonium nitrate above 5 g/liter inhibited the degradation of Dyfonate by R. arrhizus. Time-course studies on the metabolism of the insecticide indicated that Dyfonate was first absorbed by the fungal mycelium, where it was metabolized followed by the release of water-soluble, noninsecticidal, breakdown products into the culture media. The degradation appeared to involve the breakdown of Dyfonate into ethyl acetate soluble metabolites, such as ethylethoxyphosphonothioic acid, ethylethoxyphosphonic acid, methyl phenyl sulfoxide, and methyl phenyl sulfone. These compounds were then further degraded into water-soluble products. The optimum conditions for the degradation of the insecticide by R. arrhizus were observed at pH 6.0 to 7.0 and at 15-25 degrees C. Aged fungal mycelia were as active as mycelia in the logarithmic growth phase.     

MORPHOLOGY OF CHAETOMIUM ERRATICUM

Development of perithecia from single, uninucleate ascospores disclosed a homothallic condition for Chaetomium erraticum. This species was found to produce sessile ascogonial coil initials from uninucleate vegetative cells that become enveloped by hyphae formed at the base of the ascogonium. The ascogonium consists of several cells that are uninucleate or binucleate. A perithecium forms from numerous divisions and enlargement of the surrounding uninucleate cells. Differentiation of the perithecial cells results in the formation of a carbonaceous wall, perithecial hairs, and an ostiole lined with periphyses. A convex hymenial cluster of ascogenous cells forms in the lower half of the centrum from which typical croziers develop. Asci push up into the pseudoparenchyma cells of the centrum. The growth of the ascogenous system is in part responsible for increase in perithecial size. The breakdown of the pseudoparenchyma cells around the developing asci results in the formation of a central cavity in which ascospores are released when the asci deliquesce. No paraphyses are present. The type of development and features of the centrum of C. erraticum and other species of Chaetomium indicate a distinct Xylaria-type centrum.     

New species and new Chinese records of Bionectriaceae (Hypocreales,Ascomycota)

Collections of bionectriaceous fungi from different areas of China were examined, in which 3 new species were encountered. Bionectria intermedia is characterized by smooth perithecia, 2-layered perithecial wall, cylindrical to clavate asci with an apical ring, ascospore striations composed of separate warts, and dimorphic conidiophores. Hydropisphaera yunnanensis has hairy ascomata which are cupulate when dry, clavate asci with a simple apex, and spinulose and very narrow ascospores fusiform and constricted at septum. Nectriopsis apiosporae possesses laterally pinched perithecia when dry, 1-layered perithecial wall, rough perithecial surface, clavate asci with a simple apex, spinulose ascospores with 3 septa, and on Apiospora sp. Four species, Bionectria epichloë, B. kowhaii, B. subquaternata and Hydropisphaera suffulta, are reported as new to China.     

Selective effects of purine and pyrimidine analogues and of respiratory inhibitors on perithecial development and branching in sordaria

The initiation of perithecia in the homothallic ascomycete Sordaria fimicola was completely suppressed, without seriously inhibiting vegetative growth, by growing the fungus on an agar medium containing one of the following additions: 1) 1 μm 5-fluorouracil, 2) 10 to 100 μm 6-azauracil, 8-azaguanine or 8-azaadenine, 3) 50 to 500 μm cyanide or azide, 4) 5% (w/v) casein hydrolysate. In contrast to the selective activity of the analogues of 3 RNA bases, whose inhibition could be reversed by the appropriate normal bases only, none of the analogues of thymine were active, neither were the thio-derivatives of RNA bases. Other inhibitors of RNA and protein synthesis, like actinomycin D, puromycin and cycloheximide, were also without selective activity, although the last of these inhibited perithecial maturation at 0.1 μm concentration but not initiation. Amino acid analogues were inactive, as were the metabolic inhibitors thiourea, 2,4-dinitrophenol and fluoride. The compounds which inhibited the formation of perithecia also lowered the branching frequency of leading hyphae, but not their linear growth rates. Consequently, the branch densities were diminished in their presence. Hypotheses to account for these findings are discussed in terms of inhibition of growth in general, of the synthesis of some specific messenger RNAs, and of RNA-mediated transport across membranes, the last of which seeming the most fruitful for further work.     

Inhibition, by lanthanides, of neutral proteinases secreted by human, rheumatoid synovium

Fragments of human, rheumatoid synovium were maintained on organ culture for three days under serum-less conditions. Their conditioned media contained collagenolytic, gelatinolytic and caseinolytic activities, which were susceptible to inhibition by lanthanide ions. Of the four lanthanides tested, Sm3+ proved the best inhibitor of gelatinase and caseinase, while La3+ inhibited collagenase the most strongly. Inhibition of collagenase by La3+ was uncompetitive. A direct binding assay confirmed the greater association between collagen fibrils and collagenase in the presence of La3+. Ca2+ was not required for binding of the uninhibited enzyme to collagen, but acted to stabilize collagenase against thermoinactivation.