Amanita sinensis, new to Japan and Nepal

AnAmanita species, known as “haikagura-tengutake” in Japan and recently collected in Nepal, is identified asAmanita sinensis. Previously reported only from Southwest China, this species, distributed in Japan, Southwest China, and Nepal, is now considered to be a Southeast Asiatic element.     

An acid-tolerant lectin coupled with high Hg2+ potentiated hemagglutination enhancing property purified from Amanita hemibapha var. ochracea

A 37.4 kDa acid tolerant lectin was isolated and purified from dried fruiting bodies of Amanita hemibapha var. ochracea designated as AHL. The lectin was not adsorbed on DEAE-cellulose, but rather adsorbed on S-Sepharose and subjected to gel filtration by fast protein liquid chromatography on Superdex 75. The purified lectin was immune from inhibition activities of metal ions. More over, AHL exhibited high agglutination activity on rabbit erythrocytes with accelerating Hg²⁺ ions concentration. Partial peptide sequence analysis (VSNNLLTGPKVVR) of this lectin showed relative similarity to phosphoenolpyruvate carboxykinase [ATP]-like protein as predicted from Fragaria vesca subsp. Vesca. Interestingly, AHL displayed a strong affinity toward α-Lactose, making our study the first report associating Amanita species’ lectin specificity for α-Lactose to the best of our knowledge.     

Geographically structured host specificity is caused by the range expansions and host shifts of a symbiotic fungus

The inability to associate with local species may constrain the spread of mutualists arriving to new habitats, but the fates of introduced, microbial mutualists are largely unknown. The deadly poisonous ectomycorrhizal fungus Amanita phalloides (the death cap) is native to Europe and introduced to the East and West Coasts of North America. By cataloging host associations across the two continents, we record dramatic changes in specificity among the three ranges. On the East Coast, where the fungus is restricted in its distribution, it associates almost exclusively with pines, which are rarely hosts of A. phalloides in its native range. In California, where the fungus is widespread and locally abundant, it associates almost exclusively with oaks, mirroring the host associations observed in Europe. The most common host of the death cap in California is the endemic coast live oak (Quercus agrifolia), and the current distribution of A. phalloides appears constrained within the distribution of Q. agrifolia. In California, host shifts to native plants are also associated with a near doubling in the resources allocated to sexual reproduction and a prolonged fruiting period; mushrooms are twice as large as they are elsewhere and mushrooms are found throughout the year. Host and niche shifts are likely to shape the continuing range expansion of A. phalloides and other ectomycorrhizal fungi introduced across the world.     

白毒鹅膏菌人工驯化及毒力变异试验

我们对白毒鹅膏菌 [Amanitaverna (Bull.:Fr.)Pers.exvitt]进行组织分离获得了菌丝体纯培养 ,采用 3种栽培培养基作出菇比较试验 ,以杂木屑、麦皮、干牛粪组成的培养料菌丝生长最好 ,并发现依次为天然 (干品 )→母种→菌丝的减毒变异 ,对出菇生态作了观察 ,可见子实体原基形成 ,可惜未见出菇效应。     

β-鹅膏毒肽的反相高效液相色谱分离纯化

用反相高效液相色谱,以0.02mol/L醋酸铵-乙腈为流动相的梯度洗脱模式,在295nm吸收值的条件下,灰花纹鹅膏菌Amanitafuliginea的肽类毒素可以被成功的分离和纯化。单个肽类毒素的鉴定是用反相高效液相色谱和质谱同时进行。用这一方法可从灰花纹鹅膏菌中分离纯化出β-鹅膏毒肽(β-amanitin),产量可达到:1158靏/g(干重),产品纯度达98%以上,回收率为95.3%。β-鹅膏毒肽的分子量为919.3Da。这个方法可用于其它鹅膏菌肽类毒素的分离纯化。     

长白山鹅膏菌肽类毒素的HPLC分析

采用HPLC法对长白山地区分布的10种鹅膏菌中的-鹅膏毒肽(-amanitin)、鹅膏毒肽(-amanitin)和鬼笔毒肽(phalloidin)3种毒素的含量进行了测定。结果表明:白鹅膏(A.verna)和鳞柄白鹅膏(A.virsa)中含有-amanitin和-amanitin两种毒素,二者-amanitin的含量分别为 1861.85g/g和2477.02g/g,均高于欧洲产毒鹅膏(A.phalloides)中的含量(1607g/g)而接近灰花纹鹅膏Amanita fuliginea中的量(2633.80g/g)。毒鹅膏A.phalloides中含有3种毒素,并且菌蕾中的含量高于成熟子实体,尤其菌蕾中Phalloidin的含量(1113.35g/g)是灰花纹鹅膏成熟子实体中(432.5g/g)的3倍。     

Intensive hemodialysis and hemoperfusion treatment of Amanita mushroom poisoning

Over a period of fifteen years, 41 patients including 23 males and 18 females with Amanita mushroom poisoning were treated at the University Hospital of Lund, Sweden. The intensity of poisoning was graded according to serum transaminase elevations and prothrombin time reductions. Severity was mild in 16 patients (Group A), moderate in 14 (Group B) and severe in 11 (Group C). Members of Group C reported shorter latency periods before the onset of symptoms, (10±1 hours,p<0.05) and longer delays in treatment, (34±4 hours), than did the other patients. Intensive treatment was begun before the results of urine amatoxin assays were reported. Treatment consisted of: fluid and electrolyte replacement, oral activated charcoal and lactulose, IV penicillin, combined hemodialysis and hemoperfusion in two 8 hour sessions, some received IV thioctic acid, others IV silibinin, all received a special diet. This combination of treatment modalities was used to accelerate the elimination of amatoxin from the patients' bodies. The longest period of hospitalization, 13±2 days, was required by the patients of Group C (p<0.01). All patients improved and were discharged from the hospital asymptomatic. No sequelae were later reported for the majority of those moderately and severely poisoned. We have concluded that intensive combined treatment applied in these cases is effective in relieving patients with both moderate and severe amanitin poisoning.     

Sucrose Utilization of the Ectomycorrhizal Fungi Amanita muscaria and Hebeloma crustuliniforme Depends on the Cell Wall-bound Invertase Activity of their Host Picea abies

The role of apoplastic invertase (β-d -fructofuranoside — fructohydrolase, EC 3.2.1.26) of the host Picea abies for carbohydrate uptake and growth of two of its natural ectomycorrhiza partners was studied. For that purpose, hyphae of Amanita muscaria (Pers. ex Fries) Hock. and Hebeloma crustuliniforme (Bull. ex Fries) Quell., as well as roots and suspension cultured cells of Picea abies (L.) Karst. were used. Apoplastic invertase activity was demonstrated on roots and suspension cultured cells of spruce (in the latter case with 21.7 nkat (g fresh weight)^?1). Inhibition of the root cell wall invertase activity (pH optimum 4.5) by increasing the apoplastic pH allowed determination of the permanent release of sucrose from the root. However, under in vivo conditions at a lower cell wall pH the hydrolysation products glucose and fructose were predominantly found. In contrast to spruce cells and certain fungi, such as Saccharomyces (Novick et al., 1981) or Phycomyces (Ruiz-Herrera et al., 1989) invertase activity of the mycorrhizal fungi Hebeloma and Amanita was negligibly low. Furthermore, sucrose could not be consumed by Amanita and Hebeloma. As a consequence, cultures of these mycorrhizal fungi starved when kept on media with sucrose as sole carbohydrate source. But addition of invertase initiated hyphal growth immediately. Studies on carbohydrate uptake of host and fungal cells confirmed that the monosaccharides glucose and fructose were readily incorporated by spruce and fungal cells, with a clear preference for glucose. From these results it is suggested that apoplastic invertase activity of the host Picea abies is a precondition for the utilization of sucrose by the studied mycorrhizal fungi during the nutritional interaction of the symbiotic partners.     

Nitrate reductase activity of nonmycorrhizal Douglas-fir rootlets and of some associated mycorrhizal fungi

Douglas-fir root tips reduced nitrate at much higher rates than seven mycorrhizal fungi, which also differed between species in rate of nitrate reduction. Results are related to nitrogen nutrition of Douglas-fir.Excised, nonmycorrhizal root tips of Douglas-fir reduced nitrate at much higher rates than seven mycorrhizal fungi commonly associated with Douglas-fir. The fungi differed significantly in degree of activity:Cenococcum geophilum, Piloderma bicolor, andRhizopogon vinicolor were highest;Amanita muscaria was intermediate; andHebeloma crustuliniforme, Thelephora terrestris, andLaccaria laccata were lowest.     

A chloro amino acid from Amanita solitaria

The mushroom Amanita solitaria contains in excess of 1000 ppm 2(S)-amino-4,5-hexadienoic acid (I), 300 ppm trans-2-amino-5-chloro-4-hexenoic acid (II), and a chloride ion concentration (2000 ppm) significantly greater than that found in other basidiomycetes. I can be converted into II in hydrochloric acid, but II is not an artifact of isolation.