Direct studies of dikaryotization in Schizophyllum commune

The growth, duplication and fate of multikaryotic hyphae bearing true clamp connections, as derived from compatible matings of Schizophyllum commune, were studied by phase contrast microscopy. The nuclei (N) of multikaryotic apices maintained a near central position during hyphal growth. True clamp connection formation occurred with near synchronous mitosis followed by septal synthesis across the clamp neck and main hyphal axis. Nuclear progeny after mitosis in a hexakaryon included 6 N in the apex, 1 N in the clamp and 5 N in the penultimate cell; the solitary nucleus in the clamp later entered the penultimate cell. Similar events occurred for clamp connection formation and mitosis in the trikaryon, quadrikaryon or pentakaryon, whether in the apex or primary branches. Nuclear content of the multikaryotic apex (2 N through 10 N) had no apparent effect on the rate of individual hyphal growth. Reduction of the nuclear number in a trikaryon occurred by long-term entrappment of a solitary nucleus in the clamp and subsequent outgrowth of the dikaryotic penultimate cell. Occasionally, more than one nucleus became entrapped in the clamp cell. The ephemeral nature of the multikaryon was indicated by the fact that older cultures appeared to be exclusively dikaryotic hyphae at the colony periphery.     

Heterokaryon formation in the basidiomycete Schizophyllum commune by electrofusion of protoplasts

Summary Conditions for high frequency electrofusion of protoplasts from the basidiomycete Schizophyllum commune are described. Visual inspection revealed up to 30% of the protoplasts engaged in fusion. Using complementing nutritional mutations, nearly 7% of the regenerated protoplasts could be recovered as heterokaryotic mycelia. The method is probably equally applicable to other basidiomycetes such as Agaricus bisporus, permitting the recovery of fusion products in the absence of selection markers.     

Intergeneric hybridization between Pleurotus ostreatus and Schizophyllum commune by PEG-induced protoplast fusion

Intergeneric hybridization between Pleurotus ostreatus and Schizophyllum commune was studied using PEG-induced fusion. The fusion of protoplasts from auxotrophic mutant strains resulted in the formation of fusion hybrids in the frequencies of 3.6 to 7.3×10^–5. Most of these fusion hybrids were monokaryotic and sterile and no heterokaryosis occurred. Most fusants showed a significantly higher nuclear DNA content when compared to parental strains and no diploids (parent 1 genome plus parent 2 genome) were found. Some fusion hybrids revealed both parental fragments in nuclear and mitochondrial rDNA PCR profiles. AP-PCR (Arbitrarily-primed Polymerase Chain Reaction) fingerprints also indicated that most of the fusion products were recombinant hybrids.     

Intraspecific hybridization between Coprinus cinereus and Schizophyllum commune by PEG-induced protoplast fusion and electrofusion

Two irreversible inhibitors, iodoacetamide and diethylpyrocarbonate, were used to select intraspecific fusion products of two mushroom species, Coprinus cinereus and Schizophyllum commune. Iodoacetamide was the more suitable inhibitor because it gave a low breakage frequency and low survival rate of the cells in the inactivation experiments. Fusion-induced by polyethylene glycol and electro-fusion were compared and, under optimal conditions, gave fusion frequencies of 16.7% to 50.0% and 6.9% to 8.4%, respectively. All fusion progeny were heterokaryons (dikaryons) and had clamp connections. There were no differences in the morphology and fruiting ability of the fusion progeny and those of the heterokaryons generated from mating.     

Insoluble hydrophobin complexes in the walls of Schizophyllum commune and other filamentous fungi

Two closely related cysteine-rich hydrophobic proteins, Sc3p and Sc4p, of the basidiomycete Schizophyllum commune are developmentally regulated and associated with the walls of aerial hyphae and fruit-body hyphae. They are present in the walls as hot-SDS-insoluble complexes which can be extracted with formic acid. The hydrophobins can then be dissociated by oxidation with performic acid. However, extraction of the walls with trifluoroacetic acid results in both solubilization and dissociation of the hydrophobin complexes into monomers. This suggests that non-covalent interactions are responsible for formation of these insoluble complexes. Carboxymethylation with iodoacetic acid only occurred after reduction with DTT indicating all cysteines in the monomeric hydrophobins involved in intramolecular disulfide bridges. Abundant proteins with similar properties were found in walls from all other filamentous fungi tested, including the basidiomycetes Pleurotus ostreatus, Coprinus cinereus, Agaricus bisporus, and Phanerochaete chrysosporium, the ascomycetes Aspergillus nidulans, Neurospora crassa, and Penicillium chrysogenum, and the zygomycete Mucor mucedo.     

Isolation of a new potent cytotoxic pigment along with indigotin from the pathogenic basidiomycetous fungus Schizophyllum commune

An indole derivative, schizocommunin, was isolated along with indigotin (indigo), indirubin, isatin, and tryptanthrin, from the liquid culture medium in which a culture of Schizophyllum commune, isolated from the bronchus of a human patient with allergic bronchopulmonary mycosis, had been grown. The structure of schizocommunin was established by spectroscopic investigation. Schizocommunin showed the strong cytotoxicity against murine lymphoma cells. The assignments of the ¹H- and ¹³C-NMR signals of indigotin were also listed. This revised version was published online in June 2006 with corrections to the Cover Date.     

Chimeric pheromone receptors in the basidiomycete Schizophyllum commune

The pheromone receptor system of the basidiomycete Schizophyllum commune is capable of ligand discrimination to confer mating specificity. The pheromone receptors of the B alpha locus were investigated for ligand discrimination in a strategy of domain swapping experiments. Several altered phenotypes of chimeric receptors have been found. These include constitutive pheromone receptors which need no ligand for activation of the downstream cascade of events. In addition, receptors still dependent on ligand were identified that had altered pheromone activation profiles, including promiscuous receptors that are activated by pheromones of all nine specificities, including the former self. In addition, highly discriminative receptors were created which are activated by only two of the eight non-self-specificities. The chimeric receptors identify the last third of the receptor as the determinant for B alpha 1 specificity, whereas B alpha 2 specificity resides in noncontiguous domains covering the first and middle parts of the receptor molecule.     

Mitosis in hyphae ofSchizophyllum commune Fr.

Summary In hyphae of the basidiomyceteSchizophyllum commune Fr. mitosis occurs through a longitudinal division of a strand of chromatin, which is followed by a parallel separation of the daughter strands.     

Degradation of ferulic acid by a white rot fungus <Emphasis Type="Italic">Schizophyllum commune</Emphasis>

Summary A ubiquitous white rot fungus Schizophyllum commune was used for the first time to study the degradation of ferulic acid. Vanillic acid was observed as one of the major products of ferulic acid catabolism, with vanillin formed as an intermediate. Almost 99.9% ferulic acid with a initial concentration of 5 mM was consumed by this fungus after 16 days of incubation at 37 °C.     

Ultrastructural differences between wall apices of growing and non-growing hyphae ofSchizophyllum commune

Summary Newly synthesized chitin at the hyphal apex ofSchizophyllum commune was shown to be highly susceptible to chitinase degradation and solubilization by dilute mineral acid. With time this chitin became gradually more resistant to these treatments. With a combination of the shadow-cast technique and electron microscopic autoradiography it could be shown that this process occurred as the newly synthesized chitin moved into subapical parts of growing hyphae but also in non-growing apices which had ceased growth after incorporation of theN-acetyl[6-³H]glucosamine. These results are in agreement with a model which explains apical morphogenesis by assuming that the newly synthesized wall material at the apex is plastic due to the presence of individual polymer chains but becomes rigidified because of subsequent physical and chemical changes involving these polymers.Dedicated to Dr. A.Quispel, Professor of Botany at the University of Leiden, on occasion of his retirement.     

Recovery of degenerate schizostatin production by screening single-basidiospore isolates inSchizophyllum commune

To recover degenerate schizostatin production in aSchizophyllum commune isolate, SANK 17785, we examined schizostatin production of 30 single-basidiospore isolates obtained from a basidiocarp of SANK 17785. One of the isolates showed high productivity (136–154 μg/ml) and maintained its high productivity for 26 mo. To explore other isolates with high schizostatin production, 76 single-basidiospore isolates from 7 wild basidiocarps were obtained and their schizostatin production was quantified. Four of the isolates produced more schizostatin than SANK 17785 did originally.     

Temporal accumulation of mannitol and arabitol in Geotrichum candidum

The temporal depletion and accumulation of polyols were investigated in the fungus Geotrichum candidum. The major intracellular polyols were tentatively identified by paper chromatography as mannitol and arabitol. Inositol was also present in small quantities, and trehalose was also detected in appreciable concentrations.Germination and vegetative growth depended on the type and concentration of the sole exogenous carbon source. Mannitol occurred in arthrospores at 9.4% of the dry weight after several days growth in 2% (w/v) glucose solid medium, and became depleted during germination and vegetative growth in liquid medium containing 2% (w/v) glucose, 2% (w/v) sodium acetate or 25% (w/v) glucose as sole carbon source. This hexitol latter accumulated during arthrosporulation. The depletion and accumulation of ethanol-soluble carbohydrate believed to be primarily trehalose was temporally similar to that of mannitol. Arabitol accumulated intracellularly during germination and vegetative growth in sodium acetate medium and 25% glucose medium. This pentitol was not detected intracellularly at any culture age during growth in 2% glucose medium.Prolonged incubation of the culture in 25% glucose medium after stationary phase was reached resulted in the gradual disappearance of arabitol from the arthrospores simultaneously with an increase in intracellular mannitol. In comparison, ethanol-soluble carbohydrate did not change with prolonged incubation in this medium.     

Two morphological markers indicating dikaryosis in Schizophyllum commune

Summary In the wood destroying basidiomycete Schizophyllum commune a method is described to recognize the onset of dikaryosis rapidly in using recessive genetic markers. The gene ai ⁺/ai causes in its mutant recessive allele (ai) the production of dark coloured fruit bodies. This can be made use of to evaluate macroscopically the formation of a dikaryon. Another useful marker is the gene rd ⁺/rd. The recessive allele (rd) causes phenotypically the formation of a round looking mycelium instead of the fringed looking mycelium, the wild type. This genetic marker which is closely linked to the A-incompatibility factor is therefore also qualified to detect the onset of dikaryosis without much effort.     

The Penicillium commune Thom and Penicillium clavigerum Demelius Fungi Producing Fumigaclavines A and B

The type strains Penicillium clavigerum VKM F-447 and P. commune VKM F-3233 are found to produce fumigaclavines A and B. Of the seven other strains of these species, only two strains, P. commune VKM F-3088 and F-3491, possess the ability to synthesize these alkaloids. It is suggested that the five other strains under study either lost such an ability or require very specific conditions for the synthesis of these alkaloids.     

Co-production of Schizophyllan and Arabinoxylan from Corn Fiber

Schizophyllum commune strain ATCC 38548 grew well on a medium containing alkaline H₂O₂-pretreated corn fiber as a sole carbon source, and clarified the culture medium within 7 days. The strain preferentially utilized the starch component of corn fiber for growth and production of schizophyllan. Culture supernatants contained approx. 50 mg schizophyllan and 200 mg arabinoxylan per g corn fiber. These polysaccharides were recovered separately by differential precipitation with ethanol. USDA - Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture. Revisions requested 29 November 2005; Revisions received 24 January 2006     

“海油雪”的形成机制及其生态效应

2010年深水地平线事故发生后,在被石油污染的墨西哥湾观察到大量的"海油雪"形成,海油雪的相关研究成为人们关注的焦点。海油雪是指石油、浮游植物、细菌黏液等组成的团聚物,能够将石油从海面沉降至海底,对石油的风化过程产生深远影响。因此,研究海油雪的形成机制和生态效应,对于深入认识海油雪在石油-海洋系统中的作用具有重要意义。本文从物理凝聚、微生物和石油分散剂三个方面对海油雪的形成机制展开探讨,分析了海油雪对石油风化、底栖生物毒性和其他污染物迁移转化的影响,并结合现有研究进行了展望。     

Enzyme activities in Chrysosporium strains

390 strains of Chrysosporium were screened for their ability to produce enzymes. All strains produced: catalase, phosphatase, lipase, amylase, DNAse and phosphoamidase. No strains showed: valine arylamidase, oxidase, -galactosidase, urease, pectolase, protease nor RNAse.     

Regions in theZ5 mating gene ofSchizophyllum commune involved in Y-Z binding and recognition

The A mating locus of the woodrotting fungusSchizophyllum commune encodes two multiallelic genes,Y andZ, which regulate the A-pathway of development. TheY alleles contain a homeobox, suggesting that the Y proteins may be DNA-binding regulatory proteins. During mating, development is induced when Y from one mating partner interacts with Z from the other mating partner; self combinations of Y and Z are inactive. Two-hybrid analyses indicate that nonself combinations of Y and Z form heteromultimers and self combinations do not. To understand Y-Z binding and self- nonself recognition further we used mutagenesis and chimeras to identify regions in one allele ofZ(Z5) that are involved in these processes. Here we report the results, which broadly define regions in Z5 that are essential for activity, Y-Z binding and Z5 allelic specificity.The sequence reported in this paper has been deposited in the Genbank database under accession number U22049