几种外生菌根真菌培养特性的研究

对 4种外生菌根真菌的培养特性进行了研究。结果表明 ,PDMA和PD培养基是培养外生菌根真菌的较适合的培养基 ;木屑、麸皮、米糠、玉米粉、蛭石是培养菌根菌菌剂的较适宜的基质 ;用MMN营养液拌料以及适宜的水分供给都有利于外生菌根真菌的生长。     

油松根际外生菌根真菌龟裂秃马勃的分离鉴定及培养

该研究采用组织分离法对黄土高原子午岭林区连家砭林场油松根际外生菌根真菌进行了分离,并采用ITS序列对分离的真菌进行鉴定,研究了菌根真菌的培养条件,为菌剂的生产奠定基础。结果表明:子午岭林区油松根际分离培养出的真菌为龟裂秃马勃(Handkea utriformis),能够与油松形成外生菌根。采用MMN培养基为基础培养基,纯培养条件下,龟裂秃马勃生长的最适pH为6.0,最适温度为25℃。以蔗糖为唯一碳源的培养基中龟裂秃马勃生长速度最快,其次是麦芽糖和葡萄糖,生长最慢的为乳糖,在以几丁质和碳酸氢钠为碳源的培养基上不生长。有机氮源中,对脯氨酸为唯一氮源的利用能力最好,其次是酪蛋白,尿素最差;无机氮源中,以磷酸氢二铵为唯一氮源时生长最好,其次是硝酸钙,最差为酒石酸铵。该研究首次报道了黄土高原子午岭林区外生菌根真菌龟裂秃马勃,并初步确定了其最佳培养条件。     

净月潭国家森林公园针叶林根系共生真菌初步调查

采用菌根形态学及分子生物学手段对采自净月潭国家森林公园的红松Pinus koraiensis、落叶松Larix gmelinii和云杉Picea asperata根系共生真菌进行了初步调查。结果表明:外生菌根形态依据不同树种表现不同,红松外生菌根主要表现为淡黄色到褐色,不分支、偶有珊瑚状分支,外延菌丝丰富或浓密;云杉菌根颜色基本为淡黄色到黄色,末端膨大或等粗,表现为弯曲状或蜿蜒状,不分支或不规则分支、偶有羽状分支,外延菌丝少;落叶松菌根淡黄色到黄色,偶有红褐色,不分支、羽状分支或不规则分支,外延菌丝浓密,也有表现外延菌丝少量。共鉴定获得共生真菌9科9属,其中红松根系外生菌根真菌2种:美洲乳牛肝菌Suillus americanus和外生菌根真菌未定种1,内生真菌3种:Phialocephala fortinii,Cylindrocarpon pauciseptatum和Phialocephala helvetica;云杉根系外生菌根真菌5种:臭红菇Russula foetens、外生菌根真菌未定种2和外生菌根真菌未定种3,Tomentella sp.1和Tomentella sp.2;落叶松根系外生菌根真菌4种:平凹盘菌Peziza depressa、拟篦边红菇Russula pectinatoides、外生菌根真菌未定种4和外生菌根真菌未定种5。该研究结果丰富了针叶树根系共生真菌的多样性,可为进一步开展菌根形态解剖学以及菌根生态学等研究提供依据。     

几种松树外生菌根真菌的研究

早在1956年前我们曾对松树和柞树的外生菌根真菌进行了分离。当时获得3株菌根真菌的纯培养,经初步鉴定属牛肝菌属(Boletus)、乳菇属(Lactarius)和滑菇属(Hebeloma)。对油松(Pinus tabulaeformis)幼苗回接试验取得了成功。在樟子松(Pimus sylvestris)幼苗的接种试验中,也获得了较好的结果;菌根化促进了樟子松幼苗的成活(张宪武等,1964)。 1978年以来,我们对外生菌根真菌又进行了研究。从不同林分下采集到能形成菌根的真菌子实体,在无菌条件下用切割组织法以分离菌根真菌,获得了纯培养菌株,根据子实体的特征进行了鉴定(邓叔群,1963)。同时对培养基选择及幼苗接种效果也进行了试验。本文是我们研究的初步结果。     

山杨苗木的菌根类型及对苗木促生作用的研究*

用Glomus mosseae,G. intraradices,G. versiforme,Sclerocystis sinuosa和Pisolithus tinctorius,Cortinarius russus,Russula foetens,Lactarius insulsus等丛枝菌根真菌和外生菌根真菌各4种,对盆钵播种的山杨实生苗进行接种试验,结果表明,4 种丛枝菌根真菌对山杨苗木均未发生侵染,与野外调查结果相符,与报道的杨属既有外生菌根又有丛枝菌根相悖。因此认为山杨属于非丛枝菌根类型。4 种外生菌根真菌均与山杨苗木合成菌根,且促生作用明显。其中C. russus最好,苗高、地径、侧根数及整株干物重增长率分别为38.13%、20.27%、70.97%和33.39%。研究结果表明,山杨属外生菌根类型。     

内蒙古地区白桦外生菌根形态类型及分子鉴定

以内蒙古不同地区白桦外生菌根为材料,采用形态解剖学方法和分子生物学手段对与白桦共生的外生菌根真菌多样性进行全面的调查,并经对所测序列与GenBank和Database比对。结果显示:在内蒙古地区与白桦共生的外生菌根真菌共13种,其中担子菌7种,子囊菌4种,分别来自于丝膜菌属、丝盖伞属、蜡壳耳属、毛革菌属、滑菇属和空团菌属、块菌属、地怀菌属。其中,菌根类型T₈和T₁₁未能提出其总DNA,根据其外生菌根形态类型并参照Agerer体视显微镜菌根图谱和Haug菌根图谱进行比较,分类鉴定为荷顿氏疣柄牛肝菌和白桦外生菌根真菌一种。结果表明,内蒙古地区白桦外生菌根真菌多样性相对较高,且与利用地上子实体鉴定的外生菌根真菌种类有一定的区别。     

内蒙古不同气候带白桦外生菌根真菌群落结构及影响因素

白桦是天然次生林生态系统演替过程中的先锋树种,是典型的外生菌根树种。基于Illumina Miseq高通量测序技术及生物信息学方法,分析了白桦外生菌根真菌多样性和群落结构及其驱动因素。结果表明,白桦根围土壤中共获得261个外生菌根真菌OTUs,隶属于2门、4纲、11目、29科、50属。在门水平上,担子菌门(Basidiomycota)真菌相对丰度占93.5%;其余为子囊菌门(Ascomycota)真菌,相对丰度占6.5%。在属水平上,相对丰度≥1%的优势属共有16个,其中红菇属(Russula,32.1%)、蜡壳耳属(Sebacina,19.9%)和丝膜菌属(Cortinarius,11.8%)的相对丰度最高,分布广泛,在所有样点均有出现。采用RDA和SPSS分析了外生菌根真菌群落组成差异的驱动因素,pH值、总氮、土壤含水量和速效钾对外生菌根真菌的分布均有极显著影响,其中,pH值对外生菌根真菌多样性及群落结构变化的解释度最高,为34.3%,其次为总氮、土壤含水量和速效钾,解释度分别为25.1%、16.8%和5.4%。研究结果阐明了跨气候带白桦外生菌根真菌的群落结构和影响因素,可为深入理...     

外生菌根真菌的分离和培养条件的研究

早在1956年,我们已对松树和柞树的外生菌根真菌进行了分离。当时获得三株菌根真菌的纯培养,经初步鉴定分属于牛肝菌属(Boletus)、乳菇属(Lactarius)和滑菇属(Hebeloma)。回接试验取得了成功。在樟子松(Pinus sylvestris)幼苗的接种试验中,也获得了较好的结果;菌根化促进了樟子松幼苗的成活。但由于我们采用的分离方法是表面灭菌法,直接从松树的小根上来分离菌根真菌     

《菌物学报》“菌根真菌专刊”征稿通知

<正>真菌与植物关系密切、共同发展、协同进化,在自然界长期演化过程中,真菌与植物逐渐建立了共生关系。业已证实,菌根真菌与植物根系建立的共生体是生物圈内典型的互惠共生关系。菌根真菌几乎涉及真菌界所有门类。其中,丛枝菌根真菌(AMF)属于专性活体营养的共生真菌,已分离鉴定近300种,隶属球囊菌门1纲4目11科25属;外生菌根真菌隶属担子菌门、子囊菌门、接合菌门和无性态真菌,全球约5 000~6 000种,     

呼伦贝尔沙地樟子松外生菌根真菌多样性

沙地樟子松是我国北方重要的防风固沙造林树种,也是一种典型的外生菌根依赖型树种。为揭示呼伦贝尔沙地樟子松外生菌根真菌多样性,以中龄、近熟、成熟3个龄组沙地樟子松人工林和沙地樟子松天然林为研究对象,采用野外调查和分子生物学相结合的研究方法,鉴定分析沙地樟子松外生菌根真菌种群特征。研究结果表明:(1)呼伦贝尔沙地樟子松外生菌根真菌共有10个OTU属于子囊菌,48个OTU属于担子菌,隶属于21科25属。(2)天然林优势菌为糙缘腺革菌属Amphinema、丝膜菌属Cortinarius和乳牛肝菌属Suillus,人工林优势菌为乳牛肝菌属,其余菌种相对丰度随着林龄变化波动较大。(3)天然林与人工林外生菌根真菌种群Shannon、Simpson和Pielou指数存在显著差异(P0.05),人工林间alpha多样性指数差异不显著(P0.05)。(4)呼伦贝尔沙地樟子松天然林和人工林外生菌根真菌种群组成存在较大差异,其中近熟林的外生菌根真菌群落组成与天然林的最为接近。     

东北主要林区针叶林下外生菌根真菌及生态分布*

采用标准地调查和路线调查相结合的方法,我们在长白山、小兴安岭、大兴安岭主要针叶林内,对岳桦-云杉林等12个主要针叶林型的外生菌根真菌进行了调查。经对采集标本整理鉴定,计有19科43属163种外生菌根真菌。其中长白山18科39属135种;小兴安岭16科35属116种;大兴岭11科22属50余种。它们在各林型下的分布有一定的规律性,其种群组成和分布的多度与林木的组成、土壤和地形条件,如海拔高度、坡向和坡度等关系密切。     

Viability of Selected Microorganisms in Hydrocarbon Fuels

A laboratory study of the viability of selected microorganisms in a hydrocarbon fuel medium was carried out on 19 species of microorganisms, representative of the types found as natural contaminants in aircraft fuels. More species remained viable when inoculated in pure cultures than when inoculated in mixed (composite) cultures. Of the 19 species selected, 10 were still viable after 3 months and 5 were viable after 4 months in the pure culture inoculants. In the complete composite culture inoculant, the bacterial species which were viable at the end of 4 months were the same as those found in the pure culture inoculant. No fungi remained viable in the complete composite cultures after a 3-week period. The microorganisms which remain viable in a hydrocarbon fuel medium are considered indicative of a satisfactory inoculum to be used as a test culture in laboratory analysis of mechanical control techniques.     

构树可培养内生真菌的多样性初探

构树Broussonetia papyrifera为桑科重要的资源植物。为调查逆境构树内生真菌的多样性,我们对铅锌矿区构树根、茎、叶和果实各部位进行内生真菌分离鉴定,从中分离得到187株内生真菌,根据形态学与分子生物学方法鉴定为20属,其中链格孢属Alternaria（50.27%）、镰孢属Fusarium（11.76%）为优势属。不同培养基的内生真菌分离率结果表明,马铃薯葡萄糖琼脂培养基（PDA）分离出的菌株数最多,占总数的73.96%;构树不同部位的菌株分离数目显示,叶中分离出的菌株较多,占38.02%。在属的分类水平上,根、茎、叶、果实不同部位的真菌数量也存在明显差异。Shannon-Wiener多样性指数和Simpson多样性指数分析结果提示不同样地的构树内生真菌的多样性也存在差异。Chao1指数和ACE指数也提示构树内生真菌丰富度与样地和组织有关。     

Ability of ectomycorrhizal fungi to utilize starch and related substrates

Basidiomycetous fungi of 55 strains of 33 species in 15 genera which are thought to be ectomycorrhizal were grown on starch and related substrates as a sole carbon source, and their ability to utilize these substrates was determined. Mycellial weights of the fungi grown on agar media containing starch and amylose varied between the strains from 1.1 to 94.9 mg/flask and from 0.4 to 93.3mg/flask, respectively. Mycelial growth rates ranged from 0 to 1.17 mm/d on barley grain medium and from 0 to 2.03 mm/d on rice grain medium; the highest rate corresponded to about half of the average of reference wood-rotting fungi. Most of the mycorrhizal fungi that grew well on amylose gave higher growth rates on barley. Several strains inLyophyllum, Hebeloma, Sarcodon, andTricholoma grew well on both glucose and starch media.     

Topography-specific emergence of fungal fruiting bodies in warm temperate evergreen broad-leaved forests on Yakushima Island,Japan

We conducted line route censuses of fungal fruiting bodies from August to September in 2005 and 2006 along ridges and valleys and compared the differences in the encounter rates of fungal fruiting bodies (= fruiting bodies seen per census kilometer) between types of topography and between fungal functional groups (i.e., ectomycorrhizal and saprobic fungi) in warm temperate evergreen broad-leaved forests on Yakushima Island, Japan. We found 251 fungal fruiting bodies (26 families, 50 genera, and 65 species) in total, including 51 bodies from Tricholomataceae, 41 from Russulaceae, 25 from Boletaceae, and 19 from Amanitaceae. The encounter rate of ectomycorrhizal fungi was greater at the ridge route (26.7 unit/km) than at the valley route (8.7 unit/km) and that of saprobic fungi was greater at the valley route (25.0 unit/km) than at the ridge route (12.5 unit/km). In addition, we conducted 7-year intermittent sampling and identified 40 families, 96 genera, and 142 species. The topography-specific emergence pattern of the intermittent sampling method was similar to that of the line census method. The fungal species composition in this study was possibly affected by a topographic gradient for both fungal functional groups through soil moisture, nutrient availability, and host tree distribution.     

Performance of Selective and Differential Media in the Primary Isolation of Yeasts from Different Biological Samples

In view of the increase in yeast infections, especially polymicrobial ones, differential culture media have acquired increasing importance. The present study evaluated the Sabouraud chloramphenicol, Biggy agar, Pagano Levin agar and CHROMagar Candida media in terms of isolation, number of yeast colony forming units per plate, and inhibition of bacteria and filamentous fungi. To this end, we used 223 biological samples, including feces, and oral, vaginal and anal mucosae from 86 patients presenting or not symptoms of fungal infections. The four media did not differ significantly in terms of detection of yeast-positive cultures. The number of colony forming units per plate ranged from zero to 2.380, with a predominance of counts of 1 to 9 colonies per plate. No significant differences were observed among the four culture media in terms of number of colonies counted, for each kind of biological material. Fifteen species belonging to the genera Candida, Saccharomyces, Cryptococcus, Trichosporon and Rhodotorula were isolated, with C. albicans being the predominant species, followed by C. parapsilosis and R. rubra. CHROMagar Candida and Biggy agar were complementary in the isolation of the different species and favored a greater recovery of polymicrobial cultures. Pagano Levin agar isolated the smallest variety of species. Sabouraud chloramphenicol agar was the least effective in terms of bacterial inhibition and favored a greater development of filamentous fungi. The results suggest that more than one culture medium should be used for an adequate primary isolation.     

姜黄根茎内生真菌区系分析

目的了解姜黄植物根茎中内生真菌群落组成和生态分布规律。方法采用表面消毒法分别于春、冬季从姜黄植物根茎中分离内生真菌,通过形态特征和基于ITS序列的系统发育分析初步确定其分类地位。结果分离获得51株内生真菌(春季33株、冬季18株),初步鉴定其分别归于5个纲、7个目、8个科、8个属(Fusarium,Gibberel-la,Alternaria,Phomopsis,Diaporthe,Nectria,Botryosphaeria,Mucor),其中镰刀菌属(Fusarium,51.0%)和赤霉属(Gibberel-la,17.6%)为优势菌群;春季分离的内生真菌分属于7个属(分离率为82.5%),而冬季的内生真菌仅归入4个属(分离率为45.0%)。结论姜黄植物中内生真菌具有较丰富的物种和系统发育多样性,在类群组成和分布上存在季节性差异,某些内生真菌(Fusariumsp.)具有宿主偏好性。     

Maintaining cultures of ectomycorrhizal and plant pathogenic fungi in sterile water cold storage.

Mycelial cultures of 64 isolates of 14 species of ectomycorrhizal fungi and 27 isolates of 15 species of plant pathogenic fungi were grown on agar medium in Petri dishes. Mycelial discs, 8 mm in diameter, were removed from the cultures and stored in sterile distilled water in test tubes at 5 degrees C. Sixty-four, 61, and 41 isolates of the symbiotic fungi were viable after 1, 2, and 3 years storage respectively. Only 19, 10, and 8 isolates of the pathogenic fungi were viable after 1, 2, and 3 years storage, respectively. Time in pure culture before water storage did not affect viability of any fungal species following water storage. After 3 years storage, four fungi (three symbionts and one pathogen) were tested and found to have retained their original growth rates and root-infecting abilities on pine seedlings. The same four isolates, however, maintained on agar slants at 5 degrees C and subcultured every 4 to 6 months, grew slower and did not infect as many feeder roots of pine as the water-stored isolates.     

未培养真菌分离培养方法初探

据估计自然界中真菌有220万到380万种,目前已经描述的真菌仅约12万种,不超过总数的8%。大量基于高通量测序的研究显示,自然环境中蕴藏的真菌多样性可能远远超出我们的预估。然而基于传统的分离培养技术的研究中,大量真菌却因难以获得纯培养而未被认知。因此探索新的真菌分离技术有助于提高我们对自然界中真菌多样性的认识,并获得可供开发利用的全新生物遗传资源。本研究以淡水湖底泥为调查对象,从优化培养条件和原位培养两个方面探索未培养真菌的分离培养方法,并与传统培养方法及免培养的高通量测序结果比较,评估各方法的分离效果。结果显示,低温分离显著影响获得的真菌组成,有利于嗜冷真菌的获得;无论在4℃低温还是25℃常温条件下,在培养基中添加维生素都能显著提高分离获得的真菌多样性,在属级水平上提高比例分别高达207%和81%。相较于传统25℃稀释平板法,基于分离芯片技术的原位培养在分离纯化效率、未知真菌捕获率以及物种多样性和均匀度等方面具有显著优势,显示原位培养技术在未来真菌分离培养中可能具有极大应用前景。     

云南松(Pinus yunnanensis)林外生菌根真菌的时空分布

2000年至2005年,调查了滇中及其附近云南松林下外生菌根真菌的生态分布,共采集、鉴定标本834号,计有27科39属211种（含变种、变型）。结果表明,红菇属（Russula）、牛肝菌属（Boletus）、乳菇属（Lactarius）、乳牛肝菌属（Suillus）、口蘑属（Tricholoma）、鸡油菌属（Cantharellus）和革菌属（Thelephora）等为云南松林下的主要外生菌根菌类群。它们的发生与分布受到气候（如：气温和降水）、植被（如：林龄、林地郁闭度和草本植被）、地形特征（如：海拔、坡向和坡度）、土壤条件（如：pH值、地表腐殖质和枯枝落叶层等）和人为干扰（比如：商业化采集、林木采伐、火烧和地表物清理）诸多因素的影响。总结为如下：（1）5a的调查结果显示,云南松外生菌根菌的分布表现出季节性变化的规律;其中以每年1、2、3月份的物种多样性为最低,雨季期间急剧增加,至中夏和秋末达到顶峰,种类最为繁多。（2）在海拔1500-2100m,云南松外生菌根菌种类随着海拔的升高而逐渐增加,至顶峰后,又呈缓慢下降趋势。海拔因素不但对其物种多样性,而且对于类群的组成也具有重要的影响。特定的类群往往发生在特定海拔范围。（3）随着云南松林龄的增加,外生菌根菌呈现由少至多的演替过程。外生菌根菌多样性随云南松林生长而逐渐增加的演替方式,可能与宿主光合作用产物、根部分泌物和土壤条件的逐渐变化有关。（4）人类干扰是云南松外生菌根菌物种多样性和类群组成的主要负影响因子。大规模的商业化采集可破坏或枯竭地下菌丝体,打破各物种之间的竞争平衡,减少孢子释放影响资源再生能力,进而直接影响到子实体的产生。外生菌根菌物种多样性的减少趋势会随林木砍伐和火烧强度的增加而加剧。地表枯枝落叶层与杂草密度也会影响子实体的产生,其中枯枝落叶层的厚度与云南松外生菌根菌子实体的发生呈负相关性,而被紫茎泽兰覆盖的云南松林地内也很少会发现相应的子实体。