Glutamate metabolism in a monokaryon of Coprinus cinereus.

The possible involvement of the glutamine synthetase/glutamate synthase pathway in ammonia assimilation is reported in a monokaryon of Coprinus cinereus.     

Asexual sporulation in coprinus cinereus: structure and development of oidiophores and oidia in an amut bmut homokaryon

Polak, E., Hermann, R., Kues, U., and Aebi, M. 1997. Asexual sporulation in Coprinus cinereus: Structure and development of oidiophores and oidia in an Amut Bmut homokaryon. 22, 112-126. Coprinus cinereus strain AmutBmut is a homokaryon with mutations in both mating type loci. It produces asexual spores (oidia) in sticky liquid droplets on specialized aerial structures (oidiophores). These oidiophores have uninucleate cells and are organized as those of the monokaryon 5026 from which the strain derived. However, unlike in the monokaryon, oidiophores in strain AmutBmut are induced by light. Young oidiophores are easily detected upon light induction and the process of oidiophore development is readily followed in this strain. Fully grown oidiophores consecutively give rise to short branches (oidial hyphae) that break up into two or occasionally three uninucleate oidia (arthroconidia) until up to 200 oidia are collected at the tip of the oidiophore. Mature spores are enclosed by a mucilage and a double-layered primary cell wall with hair-like structures except for the sides of former cell attachments. In a summary of our microscopic observations on developing oidiophores and nuclear stainings we present a model showing the successive steps of oidiophore and spore development.     

香菇B交配型位点的分子遗传学结构研究I.信息素受体和信息素前体编码基因的克隆和序列分析

本研究结合简并PCR和染色体步行两种方法研究了香菇135菌株的交配型B位点的分子遗传学结构。从135菌株的原生质体单核体1号菌株中获得了1个信息素受体编码基因LErcb1-B1和1个信息素前体编码基因LEphb1-B1。经序列比对分析,香菇的信息素受体LErcb1-B1序列与灰盖鬼伞和裂褶菌的信息素受体之间具有同源性,经SOSUI软件分析该序列具有7次跨膜结构特征。信息素前体LEphb1-B1具有CaaX基序特征。     

Characteristics of a laccase over-secreting mutant of Coprinus congregatus

Abstract Coprinus congregatus has a membrane-associated laccase which is not secreted into the culture media. A mutant monokaryon has been generated by UV irradiation which shows slower radial growth than the wild-type (72.7%) but which secretes laccase. The laccase of the mutant was compared with that of wild-type by native PAGE analysis, and showed identical mobility.     

Evidence for synthesis de novo of NADP-linked glutamate dehydrogenase in Coprinus mycelia grown in nitrogen-free medium

Abstract NADP-linked glutamate dehydrogenase is developmentally regulated in the mushroom fruit bodies of Coprinus cinereus . The enzyme can also be specifically induced by transfer of mycelium of the monokaryon BC9/6,6 to an ammonium-free medium containing an excess of pyruvate. Tests involving differential labelling show that the enzyme is synthesised de novo in this latter type of experiment.     

Purification, characterization and evaluation of extracellular peroxidase from two Coprinus species for aqueous phenol treatment

Non-ligninolytic fungal peroxidases produced by Coprinus cinereus UAMH 4103 and Coprinus sp. UAMH 10067 were purified, characterized and evaluated as cost-effective alternatives to horseradish peroxidase for aqueous phenol treatment. Purified Coprinus peroxidases exhibited a molecular weight of 36 kDa on matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Although the catalytic properties of the two Coprinus peroxidases were nearly identical in both crude and purified forms, the stabilities were substantially different. The peroxidase from Coprinus sp. UAMH 10067 was more stable at 50 degrees C and under basic conditions (up to pH 10) than the enzyme from C. cinereus UAMH 4103. The former enzyme also performed better at pH 9 than the latter one in aqueous phenol treatment. The phenol removal efficiency of the Coprinus peroxidase was comparable to those of previously studied plant peroxidases. The broader working pH and higher thermal and alkaline stability of the peroxidase from Coprinus sp. UAMH 10067 may be advantageous for its application to industrial wastewater treatment.     

Optimization of extracellular fungal peroxidase production by 2 Coprinus species

Optimum culture conditions for the batch production of extracellular peroxidase by Coprinus cinereus UAMH 4103 and Coprinus sp. UAMH 10067 were explored using 2 statistical experimental designs, including 2-level, 7-factor fractional factorial design and 2-factor central composite design. Of the 7 factors examined in the screening study, the concentrations of carbon (glucose) and nitrogen (peptone or casitone) sources showed significant effects on the peroxidase production by Coprinus sp. UAMH 10067. The optimum glucose and peptone concentrations were determined as 2.7% and 0.8% for Coprinus sp. UAMH 10067, and 2.9% and 1.4% for C. cinereus UAMH 4103, respectively. Under the optimized culture condition the maximum peroxidase activity achieved in this study was 34.5 U x mL(-1) for Coprinus sp. UAMH 10067 and 68.0 U x mL(-1) for C. cinereus UAMH 4103, more than 2-fold higher than the results of previous studies.     

Extracellular peroxidase production by Coprinus species from urea-treated soil

Thirteen strains of inky-cap mushroom Coprinus species were evaluated for the production of extracellular peroxidase. The liquid fermentation was carried out in shake flasks containing 1% glucose, 0.5% peptone, 0.3% yeast extract, and 0.3% malt extract broth at 25 degrees C. Peroxidase activity was detected in the liquid culture of several Coprinus species, including C. echinosporus NBRC 30630; C. macrocephalus NBRC 30117; Coprinus spp. UAMH 10065, UAMH 10066, UAMH 10067, and 074, after 10 days of growth. Peroxidase production by Coprinus sp. UAMH 10067, a Coprinus species isolated from urea-treated soil, was comparable to that of C. cinereus and reached 15 U.mL(-1) after 10 days. In addition, the peroxidase from Coprinus sp. UAMH 10067 was apparently more thermally stable than the enzyme produced by C. cinereus.     

The A mating-type genes of the mushroom Coprinus cinereus are not differentially transcribed in monokaryons and dikaryons

Summary The A mating type factor of Coprinus cinereus regulates part of a developmental sequence that leads to the conversion of the asexual monokaryon into the fertile dikaryon. The A42 factor is a complex of seven genes, at least four of which are involved in determining the specificity of mating interactions. In this report we show that the A42 genes are constitutively expressed in both monokaryons and dikaryons. This has important implications with respect to intracellular recognition of a compatible mating, which requires an interaction between proteins already present within the cells of the mating partners, and for the subsequent maintenance of dikaryotic growth.     

乙烯厂循环水污染物的微生物种类鉴定及其防治

某乙烯厂循环水一度出现大量性质不明的絮状污染物 ,严重影响了乙烯厂的正常生产。经鉴定 ,该污染物主要由芽孢杆菌 (Bacillussp .)和曲霉属 (Aspergillussp .)、毛霉属 (Mucorsp .)、青霉属 (Penicilliumsp .)、交链孢属(Alternariasp .)、鬼伞属 (Coprinussp .)的真菌菌丝体组成。在工厂循环水中发现有鬼伞属的真菌菌丝 ,这在其它文献中尚未见报道。针对这种絮状污染物的主要组成成分 ,广东省微生物研究所研制了能有效防控这些微生物污染的杀菌剂。     

Spore formation and karyological characterization of basidiosporogenesis,meiosis, post-meiotic and nuclear migration mitosis in Coprinus comatus

In Coprinus comatus, we sampled and observed different maturity stages of basidium to find the possible causes of binucleate basidiospores by microscopy using the HCI-Giemsa and DAPI staining methods. In basidium, following karyogamy, a single nucleolus was observed. We found evidence suggesting post-meiotic mitosis following nuclear migration in the spores. Post-meiotic mitosis occurred in the basidiospores, resulting in four binucleate basidiospores. This indicates that the basidiospore is a monokaryon, and that C. comatus shows the same pattern of basidiospore formation as Coprinopsis cinerea. This type of nuclear behavior was defined as pattern D, one of six distinct patterns (pattern A–F) of nuclear behavior during basidiosporogenesis have been described for basidiomycetes by Campos and Costa in 2010.     

毛头鬼伞的生物活性作用

毛头鬼伞具有高蛋白、低脂肪等优良特性。毛头鬼伞中含有20种氨基酸,其中人体必需的8种氨基酸全部具备;毛头鬼伞还含有钾、钠、钙、镁、磷等元素和铁、铜、锰、锌、钼、钴等微量元素。目前,毛头鬼伞已被定为符合联合国粮农组织(FAO)和世界卫生组织(WHO)要求的,集天然、营养、保健三种功能为一体的16种珍稀食用菌之一。迄今为止,有关毛头鬼伞生物活性作用的研究还开展得很不充分,国内有少量相关报道,国外对此研究甚少。本文对毛头鬼伞的生物学作用研究现状进行综述,为临床治疗肿瘤、糖尿病、感染性疾病等提供参考。     

用CHEF凝胶电泳探测远缘担子菌间原生质体融合后的染色体变异

用电场诱导灰盖鬼伞(Coprinus cinereus Co5104,his~-)和佛罗里达侧耳(Pleurotusflorida Pf67, ade~-)两菌株的原生质体融合,获得了两类属间融合子:一类为不断发生分离的单核体,另一类为稳定的双核异核体。利用等高钳位均匀电场(CHEF)凝胶电泳技术比较融合子与两亲本的染色体长度多型性(CLP),显示出融合子的染色体变异以整条染色体的丢失为主。结合形态观察和DAPI核荧光染色的结果,推测单核融合子在胞质融合后很可能发生了核融合,但来自亲本Pf67的染色体不断丢失,最后融合子的遗传组成以来自Co5104的染色体为主;而双核融合子在质配之后可能并未发生核配,不过,来自各亲本的染色体最终均只有部分得以保留在融合子中。     

The control of meiosis progression in the fungus Coprinus cinereus by light/dark cycles

Meiosis progression in Coprinus cinereus is controlled by light/dark cycles. Light is essential to propel basidia into karyogamy and light intensity determines the timing of meiotic events. The higher the light intensities, the faster the fruiting bodies enter karyogamy. The critical period when light has this influence is between 16 and 6 h before karyogamy. The control is highly stage specific. A 3-h dark period is essential for a Java dikaryon and the Japanese A(mut)B(mut) homokaryon to enter meiotic metaphase; without it the fruit body is permanently arrested at diffused diplotene. This arrest is light intensity-dependent (>20 hlx) and temperature-dependent (e.g., 27 degrees C). The placement of the dark period is very stage specific; it has no effect when placed before karyogamy stage. A dikaryon of London origin is light blind and able to complete meiosis under continuous high light regime. Fruiting bodies arrested under a continuous high light can be rescued by a 3-h dark treatment, but there is always an 8-h lag time to enter meiotic metaphase. It is possible that the dark effect signals cellular processes leading to division events. Cytological studies of arrested fruiting bodies showed that chromosomes are normal in meiotic prophase through pachytene and diplotene, but are unable to undergo chromosome condensation. Genetic crosses between a monokaryon of Java stock J6;5.4 and a monokaryon BL55 or H5 of London stock showed that light-blindness is dominant, and is controlled by a single Mendelian gene.     

Coprinol, a new antibiotic cuparane from a Coprinus species

Coprinol, a new antibacterial cuparane, was isolated from fermentations of a Coprinus sp. Its biological activities were investigated and its structure was elucidated by spectroscopic methods. The new antibiotic exhibited activitiy against multidrug-resistant Gram-positive bacteria in vitro. Two derivatives were synthesized and their activities compared to the parent compound.     

Observations on the Regulation of Glutamate Dehydrogenase Activity in Coprinus lagopus

Extracts of the mycelium of Coprinus lagopus (sensu Buller)contain two glutamate dehydro-genases with different optimumpH values. One is assayed with nicotinamide adenine dinucleotide(NAD-GDH) and the other with nicotinamide adenine dinucleotidephosphate (NADP-GDH). Changes in specific activity of the enzymeswere investigated during the growth of both a monokaryon (H9)and a dikaryon (H9 x TC) in different media and after the transferof mycelium from one growth medium into another. In the lattercase the magnitude of the changes in enzyme activity could bealtered by modification of either the carbon or the nitrogensource in the transfer medium. It is concluded from the resultsobtained that neither glutamate nor the ammonium ion seems toregulate directly the synthesis of either enzyme. However, someof the results are in accordance with the view that a productof glucose metabolism represses the synthesis of the NAD-GDHand derepresses or induces that of the NADP-CDH and evidencethat this regulator is 2-oxoglutarate was obtained. It is alsoconcluded that the complete system of regulation must involvemore than one molecule.     

Magnetic resonance spectral characterization of the heme active site of Coprinus cinereus peroxidase

Examination of the peroxidase isolated from the inkcap Basidiomycete Coprinus cinereus shows that the 42,000-dalton enzyme contains a protoheme IX prosthetic group. Reactivity assays and the electronic absorption spectra of native Coprinus peroxidase and several of its ligand complexes indicate that this enzyme has characteristics similar to those reported for horseradish peroxidase. In this paper, we characterize the H2O2-oxidized forms of Coprinus peroxidase compounds I, II, and III by electronic absorption and magnetic resonance spectroscopies. Electron paramagnetic resonance (EPR) and nuclear magnetic resonance (NMR) studies of this Coprinus peroxidase indicate the presence of high-spin Fe(III) in the native protein and a number of differences between the heme site of Coprinus peroxidase and horseradish peroxidase. Carbon-13 (of the ferrous CO adduct) and nitrogen-15 (of the cyanide complex) NMR studies together with proton NMR studies of the native and cyanide-complexed Coprinus peroxidase are consistent with coordination of a proximal histidine ligand. The EPR spectrum of the ferrous NO complex is also reported. Protein reconstitution with deuterated hemin has facilitated the assignment of the heme methyl resonances in the proton NMR spectrum.     

The thn mutation of Schizophyllum commune, which suppresses formation of aerial hyphae, affects expression of the Sc3 hydrophobin gene.

The spontaneous and recessive mutation thn in the basidiomycete Schizophyllum commune suppresses the formation of aerial hyphae in the monokaryon and, if present as a double dose, the formation of both aerial hyphae and fruit-bodies in the dikaryon. In the monokaryon, the mutation prevents accumulation of mRNA of the Sc3 gene, and in the dikaryon it also prevents the accumulation of fruiting-specific mRNAs, including mRNAs of the Sc1 and Sc4 genes, which are homologous to the Sc3 gene. These three genes code for hydrophobins, a family of small hydrophobic cysteine-rich proteins. In the thn monokaryon, the only detectable change in synthesized proteins is the disappearance of an abundant protein of apparent Mr = 28 K from the culture medium and from the cell walls. Protein sequencing shows that this is the product of the Sc3 gene. The Sc3 hydrophobin is present in the walls of aerial hyphae as a hot-SDS-insoluble complex. Submerged hyphae excrete large amounts of the hydrophobin into the medium.     

Antifungal antibiotic of the basidiomyceteOudemansiella mucida

Nuclear ratios were studied in terminal and subterminal cells of various mycelia of the basidiomycete Oudemansiella mucida, the producer of the antifungal antibiotic mucidin (MuciderminR Spofa). The dikaryon, the monokaryon, and the mucidin-producing strain that had been cultivated for a long time under submerged conditions were compared. Dedikaryotization was found to have taken place in the producing strain. The originally dikaryotic culture with characteristic clamp connections on the mycelium and with two nuclei in every hyphal cell lost permanently the clamp connections, probably owing to continuous intense agitation. The hyphae contained solely mononuclear cells. Mating with a compatible monokaryon yielded a dikaryon capable of normal fructification.     

Sexual selection in mushroom-forming basidiomycetes

We expect that sexual selection may play an important role in the evolution of mushroom-forming basidiomycete fungi. Although these fungi do not have separate sexes, they do play female and male roles: the acceptance and the donation of a nucleus, respectively. The primary mycelium (monokaryon) of basidiomycete fungi, growing from a germinating sexual spore, is hermaphroditic, but it loses female function upon the acceptance of a second nucleus. The resulting dikaryon with two different nuclei in each cell retains a male potential as both nuclei can fertilize receptive mycelia. We tested the occurrence of sexual selection in the model species of mushroom-forming basidiomycetes, Schizophyllum commune, by pairing monokaryons with fully compatible dikaryons. In most pairings, we found a strong bias for one of the two nuclei although both were compatible with the monokaryon when paired alone. This shows that sexual selection can occur in mushroom-forming basidiomycetes. Since the winning nucleus of a dikaryon occasionally varied depending on the receiving monokaryon, we infer that sexual selection can operate through choosiness of the receiving individual (analogous to female choice). However, in other cases the same nucleus won, irrespective of the receiving monokaryon, suggesting that competition between the two nuclei of the donating mycelium (analogous to male–male competition) might also play a role.