线虫草科弯颈霉属研究进展

弯颈霉属Tolypocladium是W. Gams于1971年建立的新属,该属的特点是瓶梗基部呈球形或椭圆形膨大,瓶颈细长而弯曲,分生孢子单生或黏聚成小头状,截至目前国际真菌名录数据库中记录弯颈霉属真菌有43种。其模式种为膨大弯颈霉Tolypocladium inflatum W.Gams。弯颈霉属是一类重要虫生真菌,生态分布广,寄主种类多,弯颈霉属很多种被发现是虫草属无性型,与虫草属的关系十分密切,2014年一些国际著名真菌学家基于分子生物学的新分类系统将弯颈霉属归入到线虫草科Ophiocordicipitaceae。弯颈霉属能产生多种杀虫和抗菌的有效成分,如环孢菌素(Cyclosporin)、大团囊素(Ophiocordin,蛋白酶强抑制剂)、线肽素(efrapeptin)、四聚胺酸素(Ophiosetin)、多种杀虫剂等。文中综述了弯颈霉属真菌的分类、生活习性与生态分布等,介绍了其药理药效及有效化学成分等相关研究,并展望了弯颈霉属真菌的研究发展前景。     

高山弯颈霉(Tolypocla dium nubicola),中国弯颈霉属一新记录种

报道了分离自土壤的一个弯颈霉属新记录种 ,即高山弯颈霉 (TolypocladiumnubicolaBissett)。其主要鉴别特征是 :①在查氏培养基上 ,2 5℃ ,14d ,直径达 30～ 35mm ,平展 ;②瓶梗直接着生于菌丝或简单的分生孢子梗上 ,有再育现象 ,基部拟椭圆形膨大或球形膨大 ,(2 .6～ 7.2 ) μm× (1.3～ 3.2 ) μm ,颈部细长 ,常弯曲 ,(3.0～6 .3) μm×0 .3μm ;③分生孢子单生 ,柱状至椭圆形 ,(1.6～ 3.8) μm× (0 .8～ 1.2 ) μm ,外无黏液层。     

从长白山土壤中分离的3种弯颈霉属真菌(英文)

采用土壤稀释平板法从采自长白山自然保护区北坡森林的土壤样品中分离纯化出较多的弯颈霉属(Tolypocladium)真菌菌株,经形态观察鉴定出3个种,即雪白弯颈霉(T.niveum)、柱孢弯颈霉(T.cylindrospo-rum)及地生弯颈霉(T.geodes),其中(T.cylindrosporum)和(T.geodes)为中国新记录种。统计分析结果表明这3种真菌为喜冷真菌,表现出具有海拔分布特点。     

地生弯颈霉的培养与生长条件

从长白山北坡森林土壤中分离纯化得到1株地生弯颈霉(Tolypocladium geodes),对其培养及生长条件(如培养基、光照、温度、碳源、氮源和pH等)进行了试验。结果表明,地生弯颈霉在麦芽浸膏琼脂培养基(MEA)上菌落生长直径最大,马铃薯葡萄糖琼脂培养基(PDA)上产孢量最高;5~30℃温度范围内均能生长,25℃时生长和产孢量最佳;最适碳源和氮源分别为葡萄糖和柠檬酸铵;光暗交替条件下,最利于其生长和产生孢子;pH4~9范围内均能生长和产孢,pH=8时菌落直径最大,pH=6时产孢量最高。     

中国弯颈霉培养物的化学成份分析

中国颈霉Ｔ．ｓｉｎｅｎｓｅ是与冬虫夏草无性型密切相关的真菌。对中国弯颈霉ＺＮ９２３菌株培养物进行测试,证明其所含化学成分如甘露醇、氨基酸、生理碱、有机酸及甾醇类等与天然冬虫夏草基本相同。红外、紫外吸收图谱也表明两者所含的各类化学份极为相似。     

中国弯颈霉培养物的化学成份分析

中国颈霉T.sinense是与冬虫夏草无性型密切相关的真菌。对中国弯颈霉ZN923菌株培养物进行测试,证明其所含化学成分如甘露醇、氨基酸、生物碱、有机酸及甾醇类等与天然冬虫夏草基本相同。红外、紫外吸收图谱也表明两者所含的各类化学成份极为相似。     

中国弯颈霉的培养性状和药理作用研究

中国弯颈霉ＺＮ９２３菌株分离自蔗褐蠹蛾Ｐｈｒａｇｍａｔｏｅｃｉａ ｃａｓｔａｎｅａｅ Ｈｕｅｒｂｎｅｒ虫尸。其生长最适温度为２５￣２８℃,最适ｐＨ６￣７,在富氮培养基上菌丝生长茂盛、洁白,有香气。药理试验表明,该菌株培养物具有镇静、抗炎、增强耐氧能力、扩张气管的雄性激素样作用,急性毒性试验以８０ｇ／ｋｇ（最大允许容积）剂量对小鼠一次灌胃,未见不良作用。中国弯颈霉是与冬虫夏草Ｃｏｒｄｙｃｅｐｓ ｓｉ     

帚霉属的一个新种

从云南省哀牢山土样分离到一株帚霉属(Scopulariopsis Bainier)菌株 Y82-922,进行了它的形态学研究。它兼具瓶形和柱形并且可以层出的环痕瓶梗,分生孢子小,多为球形,直径2—3μm,有1—3个芽孔,可在原位上产生短侧分生孢子链并以其末端与亲本主链相接合。它的分类学位置介于细微帚霉(Scopulariopsis parvula Morton et Smith)与寡见拟沃德霉(Wardomycosis inopinatus Udagawa et Furuya,)之间,但具有不同于二者的特性,故定名为新种,云南帚霉 Scopulariopsts yunnanensis Chen et Jiang,sp nov.     

浙江土壤中的暗色丝孢菌研究Ⅰ.

从浙江省采集的42份土壤样品中,分离出50个暗色丝孢菌分离物,鉴定为26属39种.其中拟苍白弯孢Curvularia subpallescens为新种.侧多隔孢属Pleurophragmium,疣瓶孢属Eladia,拟葡萄孢属Pseudobotrytis和棘瓶孢属Echinobotryum为中国新记录属.鞘孢霉Chalara state of Ceratocystis adiposa,紫棕毛束霉Doratomyces purpureofuscus,黑棘瓶孢Echinobotryum atrum,小囊疣瓶孢Eladia saccula,隘缩小内多隔孢Endophragmiella constricta,青霉状粘束孢Graphium penicillioides,黑侧多隔孢Pleurophragmium actum,土栖拟葡萄孢Pseudobotrytis terrestris,腐植齿梗孢Scolecobasidium humicola和虎尾兰葡萄穗霉Stachybotrys sansevierae为中国新记录种.其余28种为国内已报道种.     

长白山分解纤维素的瓶霉属和外瓶霉属真菌

瓶霉属、外瓶霉属真菌在自然界中广泛分布,是有着重要经济意义的一类真菌。迄今,我国已报道疣状瓶霉（Phialophoraverrucosa）、裴氏瓶霉（Ph.pedrosoi）、棘状外瓶霉（Exophialaspinifera）、皮炎外瓶霉（E.dermatitidis）和甄氏外瓶霉（E.jeanselmei＝Ph．gougerotii）5种,均是分离自人体的病原真菌。在长白山自然保护区的原始林中,用生长锥取样器随机钻取腐朽林木髓心,在实验室进行分离培养。共鉴定出瓶霉属真菌2种,外瓶霉属真菌3种,其中美州瓶霉（Ph.americana、烂木瓶霉（Ph．richardsiae）和鲑外瓶霉（E.salmonis）为国内新记录种。     

Mycelium cultivation, chemical composition and antitumour activity of a Tolypocladium sp. fungus isolated from wild Cordyceps sinensis

AIMS: To examine and illustrate the morphological characteristics and growth kinetics of Cs-HK1, a Tolypocladium fungus, isolated from wild Cordyceps sinensis in solid and liquid cultures, and the major chemical constituents and antitumour effects of Cs-HK1 mycelium. METHODS AND RESULTS: The Cs-HK1 fungus was isolated from the fruiting body of a wild C. sinensis and identified as a Tolypocladium sp. fungus. It grew rapidly at 22-25 degrees C on a liquid medium containing glucose, yeast extract, peptone and major inorganic salts, with a specific growth rate of 1.1 day(-1), reaching a cell density of 23.0 g dw l(-1) in 7-9 days. Exopolysaccharides accumulated in the liquid culture to about 0.3 g l(-1) glucose equivalent. In comparison with natural C. sinensis, the fungal mycelium had similar contents of protein (11.7-microg) and carbohydrate (654.6-microg) but much higher contents of polysaccharide (244.2 mg vs 129.5 mg), adenosine (1116.8-microg vs 264.6 microg) and cordycepin (65.7 microg vs 20.8 microg) (per gram dry weight). Cyclosporin A, an antibiotic commonly produced by Tolypocladium sp., was also detected from the mycelium extract. The hot water extract of mycelium showed low cytotoxic effect on B16 melanoma cells in culture (about 25% inhibition) but significant antitumour effect in animal tests, causing 50% inhibition of B16 cell-induced tumour growth in mice. CONCLUSIONS: The Tolypocladium sp. fungus, Cs-HK1, can be easily cultivated by liquid fermentation. The mycelium biomass contained the major bioactive compounds of C. sinensis, and the mycelium extract had significant antitumour activity. SIGNIFICANCE AND IMPACT OF THE STUDY: The Cs-HK1 fungus may be a new and promising medicinal fungus and an effective and economical substitute of the wild C. sinensis for health care.     

寄生隐丛赤壳菌Cryphonectria parasitica致病毒素Cp-I的分离纯化和结构分析

寄生隐丛赤壳菌Cryphonectria parasitica菌株经液体培养,石油醚萃取其发酵液获得对板栗带叶嫩枝具有致萎活性的粗提物,以氯仿:石油醚:甲醇(6:2:2)作洗脱剂,粗提物经硅胶色谱分离,共得到3组纯组份,其中第1组份(Cp-I)对板栗幼苗致萎活性较高.质谱、核磁共振和红外光谱测定表明Cp-I分子量为278,化学式为C<,16>H<,22>O<,4>.     

寄生隐丛赤壳菌致病毒素Cp-I的分离纯化和结构分析

寄生隐丛赤壳菌Cryphonectria parasitica菌株经液体培养,石油醚萃取其发酵液获得对板栗带叶嫩枝具有致萎活性的粗提物,以氯仿:石油醚:甲醇(6:2:2)作洗脱剂,粗提物经硅胶色谱分离,共得到3组纯组份,其中第1组份(Cp-I)对板栗幼苗致萎活性较高。质谱、核磁共振和红外光谱测定表明Cp-I分子量为278,化学式为C16H22O4。     

FRESHWATER HYPHOMYCETES OF THE NORTHERN APPALACHIAN HIGHLAND INCLUDING NEW ENGLAND,AND THREE COASTAL PLAIN STATES

Twenty-seven species of freshwater Hyphomycetes from two five-state areas in the northern Appalachian Highland, New England, and three Coastal Plain states were collected, isolated, and identified. The spores of three fungi which are probably aquatic Moniliales were also studied. Sigmoidea, a new Hyphomycete genus, includes submerged aquatic fungi which produce phialides laterally along the mycelium and sigmoid, multicellular phialospores. Varicosporium giganteum sp. n. produces terminal, hyaline, branched, and multiseptate aleuriospores. The main spore axis has one to four lateral branches which arise successively on one side of this axis. Tricladium marylandicum sp. n. is similar to T. splendens in that the lateral branches are constricted where each branch joins the main spore axis. It differs from this species in that the spore has a blunt apical cell on the main axis and on each of the lateral axes. Three possible methods of dispersal are presented to explain the cosmopolitan distribution of freshwater Hyphomycetes.     

Cyclosporin A--a review on fermentative production, downstream processing and pharmacological applications

In present times, the immunosuppressants have gained considerable importance in the world market. Cyclosporin A (CyA) is a cyclic undecapeptide with a variety of biological activities including immunosuppressive, anti-inflammatory, antifungal and antiparasitic properties. CyA is produced by various types of fermentation techniques using Tolypocladium inflatum. In the present review, we discuss the biosynthetic pathway, fermentative production, downstream processing and pharmacological activities of CyA.     

Rheology and Hydrodynamic Properties of Tolypocladium inflatum Fermentation Broth and Its Simulation

A physico-chemical, two phase simulated pseudoplastic fermentation (SPF) broth was investigated in which Solka Floc cellulose fibre was used to simulate the filamentous biomass, and a mixture of 0.1% (w/v) carboxymethyl cellulose (CMC) and 0.15 M aqueous sodium chloride was used to simulate the liquid fraction of the fermentation broth. An investigation of the rheological behaviour and hydrodynamic properties of the SPF broth was carried out, and compared to both a fungal Tolypocladium inflatum fermentation broth and a CMC solution in a 50 L stirred tank bioreactor equipped with conventional Rushton turbines. The experimental data confirmed the ability of the two phase SPF broth to mimic both the T. inflatum broth bulk rheology as well as the mixing and mass transfer behaviour. In contrast, using a homogeneous CMC solution with a similar bulk rheology to simulate the fermentation resulted in a significant underestimation of the mass transfer and mixing times. The presence of the solid phase and its microstructure in the SPF broth appear to play a significant role in gas holdup and bubble size, thus leading to the different behaviours. The SPF broth seems to be a more accurate simulation fluid that can be used to predict the bioreactor mixing and mass transfer performance in filamentous fermentations, in comparison with CMC solutions used in some previous studies.     

Comparative study reveals unique features of the mycobiota in peat soils samples from Japan and Scotland

We isolated filamentous fungi from soil samples of peat layers in Aomori and Oita Prefectures in Japan and Perth and Kinross district in Scotland by a serial dilution plate technique. The mycobiota in each peat soil showed some common and characteristic features. The abundance of fungal isolates (CFU/g) from peat soil was low: about 1/3 to 1/30 compared with evergreen or coniferous forests or cultivated soil. Trichoderma or Mucorales species were scarcely observed; these fungi occupied only 3% of the total number of colonies. On the other hand, fungi such as Conioscypha and Tolypocladium that are normally isolated rather rarely were encountered at a comparatively high rate. Acremonium guillematii and Tolypocladium cylindrosporum were recorded for the first time in Japan. Sterile fungi occupied 50% of the total number of isolates. The low abundance of fast-growing fungi enabled us to pick slow-growing fungi up easily from the isolation medium. It is interesting that species not previously described in Japan, or scarcely reported, were isolated commonly from both Japanese and Scottish samples. A peat soil sample is therefore an attractive source of untapped microbial resources.     

中国弯颈霉的培养性状和药理作用研究

中国弯颈霉ZN923菌株分离自蔗褐蠹蛾PhragmatoeciacastaneaeHubner虫尸。其生长最适温度为25～28℃,最适pH6～7,在富氮培养基上菌丝生长茂盛、洁白,有香气。药理试验表明,该菌株培养物具有镇静、抗炎、增强耐缺氧能力、扩张气管和雄性激素样作用,急性毒性试验以80g／kg（最大允许容积）剂量对小鼠一次灌胃,未见不良作用。中国弯颈霉是与冬虫夏草Cordycepssinensis（Berk．）Sacc．无性型密切相关的真菌。具有潜在的药用价值。     

栎生桑黄和忍冬桑黄液体培养过程中发酵液抗氧化能力

桑黄孔菌属Sanghuangporus是药用木生真菌资源中一个重要的属,但是该属仅有少数几个种类被用于人工栽培,且栽培面积较小。此外,桑黄孔菌属中大部分种类的药用功能仍未完全明确。因此,本研究以桑黄孔菌属近期新发表的新种栎生桑黄S. quercicola和关注度较低的忍冬桑黄S. lonicericola为主要研究对象,通过测定它们液体培养过程中第2、4、6、8、10、12和14天的菌丝生物量以及发酵液的粗多糖含量、多酚含量、黄酮含量、抗坏血酸含量、DPPH自由基清除能力、ABTS自由基清除能力、总抗氧化能力、超氧化物歧化酶活性、羟自由基清除能力、超氧阴离子清除能力、铁离子还原能力和亚铁离子螯合能力等12个抗氧化指标,对桑黄的抗氧化能力进行评定。2种桑黄真菌各选取一号菌株,其发酵液均表现出强抗氧化能力。相比之下,栎生桑黄的多糖、抗坏血酸和超氧化物歧化酶含量更高,而忍冬桑黄的多酚和黄酮含量更高。相应的,栎生桑黄和忍冬桑黄在其他一些抗氧化指标上也表现出强弱程度及出现时间的差异。上述研究结果为桑黄孔菌属真菌的药用功能开发提供了新资源,为不同抗氧化代谢产物的分离纯化提供了科学依据。