Light-induced formation of fruit-bodies in a basidiomycete, Favolus arcularius (FR.) AMES

fruit-bodies in Favolus arcularius. The effect of light lastedfor about one day after transfer to darkness. The mycelium became sensitive to light about 2.5 days afterinoculation; i.e., at the beginning of the rapid growth phase.The site of fruiting was 2–5 mm inside the edge of colony(actively dividing zone) at the start of illumination. Whenone half of the plate culture was illuminated, fruiting wasrestricted to the illuminated half of the colony ; i.e., theeffect of light was localized. These results suggest that thecells sensitive to light are the actively dividing cells. Under a fixed light intensity, the total irradiation time requiredfor the initiation of fruiting was nearly constant, irrespectiveof the durations of pre-incubation in darkness and the dailyillumination period. With increasing light intensities, up toabout 500 lux, fruiting was promoted, however, a further increasein light intensity was inhibitory. (Received March 27, 1968; )     

漏斗大孔菌的化学成分研究

采用硅胶、RP-18、Sephadex LH-20等多种材料进行分离纯化,通过理化方法和波谱分析进行结构鉴定,从漏斗大孔菌发酵液的乙酸乙酯萃取部分中分离并鉴定了8个化合物,分别为:5-(hydroxymethyl)-1,1,4a-trim-ethyl-6-methylenedecahydronaphthalen-2-ol(1)、albicanol(2)、D-阿洛醇(3)、对羟基苯甲醛(4)、胡萝卜苷(5)、trili-nolein(6)、3β,5α,9α-三羟基-麦角甾-7,22-二烯-6-酮(7)和麦角甾-7,22-二烯-3-酮(8)。所有化合物均为首次从该真菌中分离得到。     

漏斗多孔菌液体发酵产漆酶条件研究

采用单因子实验分析了培养基成分及初始pH对漏斗多孔菌液体发酵产漆酶的影响。结果表明,米糠和甘蔗半纤维素组成复合碳源、NH4Cl和高C/N有利于漆酶的产生,Cu2+、米糠水解液和萘乙酸对漆酶合成具有诱导作用,1-萘酚、愈创木酚、联苯胺、乙醇、吐温80等抑制漆酶的合成,Cu2+和萘乙酸同时存在时也限制漆酶的产生,产酶培养基最适初始pH为5.2-5.7。利用优化的产酶培养基液体摇瓶培养漏斗多孔菌A08,产酶活力提高0.95倍,达到1480U/L。     

漏斗多孔菌液体菌种培养基及其栽培条件

为实现漏斗多孔菌资源化利用,设计单因素试验,以菌丝生物量、菌球密度和菌球直径为指标,获得漏斗多孔菌Polyporus arcularius液体菌种培养基配方为马铃薯(去皮)200g、玉米粉20.0g、蛋白胨5.0g、KH2PO43.0g、 K2HPO41.0g,MgSO4·7H2O 1.5g,初始pH 5.0,并优化培...     

Polyol metabolism in the mycelium and fruit-bodies during development ofFlammulina velutipes

Changes of polyol contents in the mycelium and fruit-bodies ofFlammulina velutipes were measured. The results suggested that arabinitol is accumulated in the fruit-bodies as the end-product after its translocation from the mycelium, while mannitol in the fruit-bodies is converted into fructose by the action of mannitol dehydrogenase (MDH). The development of fruit-bodies was promoted by feeding of mannitol to the mycelial colony. A¹⁴C tracer experiment indicated that half of mannitol translocated from mycelium to fruit-bodies was utilized for fruit-body development. NAD-linked MDH andd-arabinitol dehydroganase (D-ADH) were detected in both mycelium and fruit-bodies. The activities of MDH and ADH in the mycelium reached their maximum levels in the inital stage of fruit-body development and decreased thereafter. In contrast, the activity of MDH in the fruit-bodies showed a peak in the middle stage of development. The activity of ADH in the fruit-bodies was less than half of that of MDH. MDH showed a lower Km value for mannitol (1.3 ×10⁻³M) than for fructose (6.0×10⁻² M). The Km value of ADH for arabinitol was extremely high (1.3×10⁻¹M).     

Differentiation in Coprinus lagopus. III. Expansion of excised fruit-bodies.

Fruit-body expansion was studied in Coprinus lagopus (sensu Buller) following surgical procedures. Elongation occurred after denuding mushroom caps of essentially all peripheral scales. Young primordia (1 - 5 mm) failed to develop after vertical bisection. Older primordia (e.g. 10 mm) expanded 3 - 4 fold after vertical bisection or quadrisection, underwent autolysis and basidiospore production. An amorphous brown gel in the stripe lumen disappeared during development of bisected primordia. Stripes isolated from primordia expanded autonomously and exhibited negative geotropism when incubated upside down or when the stripe apex was removed. Displacement of charcoal particles dusted on intact stripes revealed the most active zone of expansion to be the upper mid-region of the stripe. Segmented stripes likewise showed most active elongation in the mid-region. Vertically bisected stripes also expanded.     

Purification and some properties of a carboxymethyl cellulase from Favolus arcularius

Favolus arcularius, a wood-rotting basidiomycete, produced carboxymethyl cellulose-hydrolyzing enzymes (CMCases) in culture media. Three main peaks of CMCase activity were separated as CMCase I, II and III at pHs from 4.4 to 5.2 by isoelectric focusing. Further, CMCase IIIa was purified from the CMCase III fraction. The molecular weight of CMCase IIIa was determined to be about 28,000 by SDS-polyacrylamide gel electrophoresis. The enzyme was not active on avicel, cellobiose and laminarin, but could randomly hydrolyze cellooligosaccharides to form G₁ and G₂ units as the end products. The apparent K_m value of the enzyme against CMC was 0.28%.     

Two new isodrimene sesquiterpenes from the fungal culture broth of Polyporus arcularius

Two new isodrimene sesquiterpene derivatives, 2(S)-hydroxyalbicanol (1, =(2S,4aS,8S,8aS)-8-(hydroxymethyl)-4,4,8a-trimethyl-7-methylenedecahydronaphthalen-2-ol) and 2(S)-hydroxyalbicanol 11-acetate (2, =((1S,4aS,7S,8aS)-7-hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)methyl acetate) were isolated from the culture broth of the fungus Polyporus arcularius, together with two phenylpropanediols, (1S,2S)- and (1R,2S)-1-phenyl-1,2-dihydroxypropane (3, 4). Compound 3 is reported as a naturally occurring compound for the first time. The structures of the compounds were elucidated on the basis of spectroscopic analysis. Compound 1 exhibited growth inhibition of lettuce seedlings with IC₅₀ values of 1.3 mM to hypocotyl and 1.7 mM to radicle.     

草菇子实体不同成熟阶段的比较蛋白质组学分析

采用iTRAQ标记结合二维液相色谱串联质谱技术对草菇不同成熟阶段的差异蛋白质组进行研究。首先将提取的草菇不同成熟阶段蛋白样品进行SDS-PAGE分析,其次将经二维液相色谱串联质谱技术获取的串联质谱数据通过MASCOT软件搜库,之后对鉴定蛋白质数据进行了KEGG代谢通路分析。试验共计获得2 335个不同肽段, 鉴定到1 039个蛋白质,其中1 030个蛋白质具有定量信息。与蛋形期相比,在伸长期和成熟期阶段显著上调蛋白质85个,下调蛋白质103个。KEGG代谢通路分析结果显示,草菇不同成熟阶段中的68个差异蛋白质可定位于4种伞菌目模式真菌（灰盖鬼伞、双色蜡蘑、可可丛枝病菌和裂褶菌）的45个不同生物代谢途径,全景展示出草菇成熟阶段差异表达蛋白质定位的代谢网络。结果表明,iTRAQ标记技术结合二维液相色谱串联质谱可对不同生长发育时期的草菇蛋白样品进行有效地分离和鉴定。     

Analysis of organic acids in fruit-bodies of Tricholoma giganteum by high performance liquid chromatography

A rapid method for determining the sensitivity of Staphylococcus aureus to beta-lactams is described. The method involves the culture of microcolonies of the bacterium on the surface of antibiotic-containing agar medium on microscope slides. The areas of the microcolonies formed are measured by means of a microscope and image analyzer. The sensitivity of the bacterium can be estimated in 4 h by this technique and the results are comparable to those obtained by overnight incubation.     

Activation of intracellular and extracellular phenol oxidases in photoinduced fruit-body formation ofFavolus arcularius

The activation of intracellular phenol oxidase (PO) was associated with the photoinduced fruit-body initiation inFavolus arcularius. The second activation of PO activity was also photoinduced in the rapidly developing stipe after the formation of pileus primordium under light exposure. However, the activity levels in the pileus and the mycelium remained low even in the cultures exposed to light. The extracellular PO activity in the culture filtrate also appears to be developmentally regulated: it rose sharply after primordium formation in the light, then decreased rapidly during further development of the fruit-body.     

Responses of Hebeloma radicosum fruit-bodies to light and gravity: negatively gravitropic and nonphototropic growth

Responses of the long-rooting agaric Hebeloma radicosum fruit-bodies to light and gravity were studied. In light from below or obliquely below, fruit-bodies grew straight downward with gills tilted and cap swollen and waved if they had emerged downward from the culture medium, or bent upward from the beginning if they had emerged obliquely downward. In light from above or obliquely above, they grew upward if they had emerged upward. Thus, they did not grow toward unilateral light from obliquely below or obliquely above, and hence their growth was nonphototropic and negatively gravitropic from the beginning of development. Even the straight downward growth seems to be latently negative-gravitropic. In the dark, fruit-bodies grew upward, forming pseudorhizas, but they remained immature; they matured only in the light. These characteristics may be related to the growth habits of the fungus colonizing deep in the ground, forming primordia there, and developing mature fruit-bodies on the ground. Received: March 26, 2001 / Accepted: July 12, 2001     

Production of fruit-bodies of a mycorrhizal fungus,Lyophyllum shimeji,in pure culture

Cultivation of mycorrhizal fungus,Lyophyllum shimeji, was examined using selected strains capable of forming primordia in pure culture. Mycelia grew fastest on barley grains containing synthetic liquid medium. The primordia readily formed in test-tubes after lowering the incubation temperature from 23°C to 15°C. The co-existence of pine seedlings had no promotive effect on primordium formation. Fruit-bodies formed on a medium consisting of barley, beech sawdust, and liquid synthetic nutrients in 500-ml glass bottles. Mature fruit-bodies produced basidiospores. The spores thus produced could germinate on an agar medium and formed mycelial colonies. Thereby, the life cycle inL. shimeji was accomplished in pure culture without using the host plant.     

超声波及脱蛋白处理对猴头菌子实体多糖提取效果的影响

本研究以猴头菌子实体为供试材料,在采用水提醇沉法进行多糖提取的过程中,系统研究了超声波和脱蛋白处理两个关键工艺环节对多糖提取效果的影响。研究结果表明：采用超声波辅助处理可提高猴头菌多糖提取率,降低多糖的纯度,使部分由蒸馏水洗脱获得的HEFP-α组分与全部0.1mol/L NaCl洗脱获得的HEFP-β组分经Sephacryl-S400洗脱后的曲线的小峰面积增加,也使主峰处的吸光值出现偏移。3种脱蛋白方法中,复合法对蛋白的除去效率最高,但多糖的保留率最低;亚铁氰化钾-乙酸锌法操作简便且蛋白除去率与多糖保留率均较高,但会使HEFP-β-2组分的含量和回收率降低;Sevage法蛋白脱除效率最低,多糖组分的保留率相对较高,且柱洗脱后发现经Sephacryl-S400凝胶柱分离后获得的主峰整体峰值较高,该方法较为适合多糖活性组分未知时的蛋白脱除。这一研究结果将会为该类大型真菌子实体的大分子生物活性物质的提取以及相关产品的开发提供重要的参考依据。     

Effect of light on the initiation of pileus formation in a basidiomycete, Favolus arcularius

Pileus formation in Favolus arcularius is induced by light,but no photoinduction occurred in young epileate stipes. Thestipes usually had to attain a length of about 5 mm to be photosensitive.Synchronous pileus formation could be induced by exposure tolight using epileate stipes which had been preincubated in darknessfor 48 to 72 hr. The pileus primordium formed about 24 hr afterthe start of illumination, however, continuous illuminationwas not necessary to produce this effect. A dark period givenbetween 1 and 8 hr after the start of illumination did not retardpileus formation. The photoinduction of pileus formation involvedtwo light-requiring processes, one occurring during the firsthour (the first light process) and the other from the 8th tothe 24th hr (the second light process). The photoresponse inthe first light process was saturated with 5 lux of light, buta light intensity below 1 lux was essentially ineffective. Onthe other hand, the reaction in the second light process couldbe started by less than 2 lux, and was accelerated by increasingthe light intensities up to about 150 lux. Further increasesin light intensity did not improve any significant effect. (Received April 30, 1974; )     

Enhancement of the viscometric endocellulase activity of Polyporus arcularius CMCase IIIa by cellobiose and cellooligosaccharides

Purification and viscometric characterization of three CMCases from Polyporus arcularius were carried out. The three CMCases, I, II, and IIIa, were estimated to have molecular masses of 39.1 kDa, 36.3 kDa, and 24.3 kDa, respectively. The addition of cellobiose and cellooligosaccharides to the reaction mixtures of CMCase I and II inhibited viscometric endocellulase activity. Following the addition of 20 mM cellobiose, CMCase I and II activities fell to about 30%–36% of their activity in the absence of cellobiose. CMCase IIIa activity, on the other hand, increased in proportion to the increase in cellobiose or cellooligo-saccharide concentration. Maximal enhancement of CMCase IIIa activity was observed following the addition of cellobiose, whereas less enhancement was observed with cellooligosaccharides spanning more than two glucoside units. The addition of 20 mM cellobiose resulted in an increase greater than 500% in CMCase IIIa activity. Inhibition of CMCase I and II by cellobiose and cellooligosaccharides may be the result of competition between the substrate and the reaction products. One of the reaction products of CMCase IIIa may bind to a site other than the active site of the enzyme, thus enhancing CMCase IIIa activity.     

Cryptoporic and isocryptoporic acids from the fungal cultures of Polyporus arcularius and P. ciliatus

In a chemical study of several fungal cultures of Polyporus, a methyl ester of cryptoporic H was isolated from P. ciliatus, together with cryptoporic acid H and 5-hydroxymethylfuran-3-carboxylic acid. Furthermore, two additional compounds, named isocryptoporic acids H and I, were isolated from P. arcularius. These isocryptoporic acids are isomers of the cryptoporic acids with drimenol instead of albicanol as the terpenoid fragment; their structural elucidation was determined by application of spectroscopic methods.     

Responses of agaric fruit-bodies to light and gravity: growth straight downward in response to light from below

Fruit-bodies of Agaricales are known to show positive phototropism during the early stage of development, but negative gravitropism at the later stage after the onset of basidiospore formation. However, when exposed to light from below, the fruit-bodies ofTephrocybe tesquorum andCoprinus spp. grew downward through all stages of development, even after the onset of basidiospore formation. Primordium formation, fruit-body development and basidiospore formation were not disturbed under such conditions. In these downward-growing fruit-bodies, gills stood straight upward. InT. tesquorum, caps often became swollen and stipes sometimes became twisted anticlockwise, contrary to those in light from above, while such behaviours were not observed inCoprinus spp.