Reserve substances inPaxillus involutus sclerotia

Summary The ectomycorrhizal fungus,Paxillus involutus, produces sclerotia in culture. These can be induced to form on agar medium by exposing mycelium grown at 25°C to various temperatures between6°C and 15°C. Sclerotia formed at 10°C and above were large and covered with drops of exudate, while those formed at 6°C or 8°C were very small and did not produce an exudate. Mature sclerotia were bounded by a compact rind and contained abundant storage reserves. Histochemistry of the larger sclerotia showed large quantities of protein stored as protein bodies in the cytoplasm, lipid present as small droplets, glycogen granules stored in the cytoplasm and polyphosphate present as small granules in the cytoplasm and in the protein bodies. Energy dispersive X-ray microanalysis confirmed the presence of phosphate in the granules and was used to map its distribution throughout the sclerotium. The smaller sclerotia induced at 8°C and below on the same medium had the same basic structure and composition, but lacked the complex phenolic cell network found in large sclerotia, and had abundant extracellular polysaccharides. The rind was not well developed and these small sclerotia are interpreted to have been arrested at an early stage of development.     

Germination and production ofMacrophomina phaseolina microsclerotia as affected by oxygen and carbon dioxide concentration

Summary Germination of microsclerotia ofMacrophomina phaseolina was observed at O₂ concentrations of 16% or higher in autoclaved soil. Germination was delayed but otherwise unaffected as O₂ decreased from 21 to 16% and was in all cases complete in 32 hours. Laboratory-produced microsclerotia consistently germinated more rapidly and seemed more independent of O₂ concentrations within the range that permitted germination than naturallyproduced microsclerotia.Population changes in soil as measured by microsclerotial counts were inversely correlated with depth of interment and reduced O₂ concentration. Our inability to detect significantly growth responses ofM. phaseolina in non autoclaved soil was apparently related to limited O₂ although other possibilities are discussed.Contribution of the Missouri Agricultural Experiment Station Scientific Journal Series No. 9124.     

外生菌根的形态学、解剖学及分类学研究进展

本文综述了外生菌根的形态学和解剖学特征,评价了这些特征用于外生菌根分类的价值,同时指出各种类型的形态学特征和解剖学特征与其营养吸收的关系。这些特征包括了菌根的颜色类型及变化、形状及分枝方式、外伸菌丝的多少及特征、根状菌索有无及分化、菌核有无及特征、菌套内外表面的菌丝排列及分化,菌套的切面特征,丹宁层厚度及分化、哈氏网的菌丝排列及厚度等。较详细阐述了100多年来,人们为外生菌根分类所作出的种种努力,并对各种分类方式作了简单的评价。     

Athelia termitophila sp. nov. is the teleomorph of the termite ball fungus Fibularhizoctonia sp.

A new species of Athelia, A. termitophila, from Japan is described and illustrated on the basis of morphological and phylogenetic analyses. Basidiomes of this species are characterized by having hyphae sometimes with clamp connections at the septa, basidia without clamp connections at the basal septa, and ellipsoid to ovoid basidiospores measuring 4.5–6 × 3–4.5 μm. In culture, mycelia produce pale brown, orange-brown to brown, globose sclerotia measuring 0.24–0.41 mm diam. The sclerotia are distinctly different in shape and size from those of other Athelia species, and are occasionally found inside the woody substrate beneath basidiomes. They are identical in shape and size to those of Fibularhizoctonia sp., also known as termite balls. Phylogenetic analysis using internal transcribed spacer (ITS) sequence data revealed that A. termitophila is the teleomorph of Fibularhizoctonia sp.     

Polyphyly,asexual reproduction and dual trophic mode in Buchwaldoboletus

Most Boletaceae form ectomycorrhizas but species in two sister groups (subfamily Chalciporoideae and the genus Pseudoboletus) have unclear trophic modes and are putatively mycoparasitic. Chalciporoideae is the earliest diverging lineage in Boletaceae and includes Chalciporus and Buchwaldoboletus. Buchwaldoboletus species fruit on wood and were thought to be saprobes, but the type species Buchwaldoboletus lignicola is mycoparasitic on the wood decay fungus Phaeolus schweinitzii. However, the nutritional mode and host range of B. lignicola and other Buchwaldoboletus remain unclear. We obtained fresh collections and axenic cultures of B. lignicola, B. hemichrysus and B. sphaerocephalus from the southeastern USA. We use cultures to examine the phylogenetic relationships among taxa in Chalciporoideae, elucidate the trophic mode of Buchwaldoboletus species and test the effect that Buchwaldoboletus species have on wood decay. Our analysis supports the hypothesis that the Chalciporus lineage is sister to all other Boletaceae but suggests that Buchwaldoboletus is not monophyletic. Confrontation experiments on agar suggest that Buchwaldoboletus species might parasitize a wide range of decay fungi. Buchwaldoboletus hemichrysus and B. sphaerocephalus produce abundant conidia in culture whereas the more distantly related B. lignicola produces sclerotia. Wood decay assays show that Buchwaldoboletus species did not have a significant effect on the decay caused by wood decay fungi. Interestingly, when B. lignicola or B. hemichrysus were inoculated alone on wood both species decayed wood, producing a brown rot that can cause 35–65% mass loss over 6 months. Our results suggest that Buchwaldoboletus species have a dual trophic mode as saprobes and mycoparasites.     

Elucidation of <Emphasis Type="Italic">veA</Emphasis>-dependent genes associated with aflatoxin and sclerotial production in <Emphasis Type="Italic">Aspergillus flavus</Emphasis> by functional genomics

The aflatoxin-producing fungi, Aspergillus flavus and A. parasiticus, form structures called sclerotia that allow for survival under adverse conditions. Deletion of the veA gene in A. flavus and A. parasiticus blocks production of aflatoxin as well as sclerotial formation. We used microarray technology to identify genes differentially expressed in wild-type veA and veA mutant strains that could be involved in aflatoxin production and sclerotial development in A. flavus. The DNA microarray analysis revealed 684 genes whose expression changed significantly over time; 136 of these were differentially expressed between the two strains including 27 genes that demonstrated a significant difference in expression both between strains and over time. A group of 115 genes showed greater expression in the wild-type than in the veA mutant strain. We identified a subgroup of veA-dependent genes that exhibited time-dependent expression profiles similar to those of known aflatoxin biosynthetic genes or that were candidates for involvement in sclerotial production in the wild type.     

Biocontrol of Sclerotinia lettuce drop by Coniothyrium minitans and Trichoderma hamatum

Fungal isolates, with known activity against Sclerotinia spp. in laboratory assays, were tested for their ability to control Sclerotinia minor in four field experiments (1998–2000). In the first experiment, eight fungal isolates (Trichoderma hamatum LU595, LU593, LU592, Trichoderma virens LU555 and LU556, Coniothyrium minitans LU112, Clonostachys rosea LU115 and Trichoderma rossicum LU596) were evaluated by incorporating spore suspensions into transplant potting mix and planting lettuce seedlings into a S. minor infested field site. At harvest, Trichoderma hamatum LU595, LU593, T. virens LU555 and C. minitans LU112 reduced disease by 30–50% compared with the untreated control under very high disease pressure (100%). In further field experiments C. minitans LU112 and T. hamatum LU593, applied as maizemeal–perlite soil amendments or incorporated into the potting mix, reduced S. minor disease over a range of disease pressures (29–91%). Disease control was equivalent or greater than that achieved with the standard carbendazim fungicide treatment. Both isolates were shown to effectively colonize the lettuce rhizosphere and surrounding soil and this colonization may have protected the roots from infection by S. minor. Multiple applications of C. minitans LU112 or T. hamatum LU593 formulations gave no added disease control compared with a single application at planting. Commercial formulations of both C. minitans LU112 and T. hamatum LU593 applied as transplant treatments, solid substrate soil amendments or as a spore drench gave consistent disease control and are currently being developed further.     

Alkaline phosphatase of <Emphasis Type="Italic">Physarum polycephalum</Emphasis> is insoluble

The plasmodia of Physarum polycephalum grow as multinucleated cells in the presence of sufficient humidity and nutriment. Under non-illuminating conditions, stresses such as low temperature or high concentrations of salts transform the plasmodia into spherules whereas dehydration induces sclerotization. Some phosphatases including protein phosphatase and acid phosphatase have been purified from the plasmodia, but alkaline phosphatase remains to be elucidated. Phosphatase of the plasmodia, spherules and sclerotia was visualized by electrophoresis gel-staining assay using 5-bromo-4-chloro-3-indolyl phosphate. Insoluble fractions of the sclerotia were abundant in phosphatase activity. The phosphatase which was extracted by nonionic detergent was subjected to column chromatography and preparative electrophoresis. Purified phosphatase showed the highest activity at pH 8.8, indicating that this enzyme belongs to alkaline phosphatase. The apparent molecular mass from sodium dodecyl sulfate-polyacrylamide gel electrophoresis under non-reducing condition was estimated to be 100 kDa whereas that under reducing was 105 kDa. An amount of 1% sodium dodecyl sulfate or 0.5 M NaCl had no effects on the activity although the phosphatase showed heat instability, Mg²⁺-dependency and sensitivity to 2-glycerophosphate or NaF. The extracting conditions and enzymatic properties suggest that this alkaline phosphatase which is in a membrane-bound form plays important roles in phosphate metabolism.     

Cytological Studies on the Process of Armillaria Mellea Infection Through the Sclerotia of Grifola Umbellata

Adhering to the sclerotium of Grifola umbellata, Armillaria mellea could invade the sclerotium in a manner of rhizomorph without capsule, after which the sclerotium formed a deep coloured stereoscopic septate cavity outside of the rhizomorph. At the early stage of infection, segmentation was visualized either in the cortex or the apex of A. mellea rhizomorph to form a new rhizomorph which penetrated another parts of G. umbellata sclerotium. At the late stage of infection, the cortical hyphae of A. mellea rhizomorph produced a branch to invade the wall of the septate cavity of G. umbellata sclerotium and, in a manner of hyphae, it could further form new rhizomorph after its penetration through that wall. An alternate way of expanding A. mellea infection in sclerotium was to form a invading band which was composed of a few rolls of round ceils derived from cortical hyphae of A. mellea rhizomorph. The band could invade sclerotia to a farther distance and then could connect with each other.