内蒙古地区山杨外生菌根形态解剖学特征研究

对内蒙古地区山杨的10种外生菌根的外部形态、颜色、分枝情况、菌索及外延菌丝的有无、菌套表面质地、是否透明、有无囊状体等结构进行了描述,结合菌套菌丝排列图式、菌套表面结构及菌丝和菌索内部结构等特征对其进行了归纳总结,并参阅有关文献进行了分类.结果表明:(1)10种菌根类型的划分基础主要是依据真菌组织学特征来加以区别,同时这10种菌根类型与其它地区对杨树外生菌根分类结果相符,符合真菌区系学分布特征.(2)树种与外生菌根共生专一性是一种双向选择模式.就山杨来说,决定其形成不同共生菌根类型的主要因子是不同环境因子影响下的真菌区系组成和树木根系发育状况.     

乳菇属真菌与松属植物形成的外生菌根

描述了两种乳菇属真菌在湿地松上形成菌根的形态学和组织学特征,结果表明两种真菌形成菌根在形态学和组织学上基本相似。其区别在于菌根的颜色及受伤后产生的乳汁的颜色。与文献报道的该属真菌形成的菌根的形态特征比较,表明具有乳汁菌丝是本属真菌形成菌根的共同特征。根据目前已有资料给出了乳菇属真菌所形成菌根的检索表。     

乳菇属真菌与松属植物形成的外生菌根

描述了两种乳菇属真菌在湿地松上形成菌根的形态学和组织学特征,结果表明两种真菌形成菌根在形态学和组织学上基本相似,其区别在菌根的颜色及受伤后产生的乳汁颜色,与文献报道的该属真菌形成的菌根的形态特征比较,表明具有乳汁菌丝是本属真菌形成菌根的共同特征,根据目前已有资料给出了乳菇属真菌所形成菌根的检索表。     

外生菌根菌在火炬松人工林应用的研究

连续６年研究了外生菌根菌在火炬松人工林的应用．结果表明,１２个供试菌株均能不同程度地在火炬松根系上形成外生菌根．在１２个供试菌株中,以松林小牛肝菌效果最佳,无论是在苗期还是上山造林,对促进寄主生长的效果均最好,且促进寄主生长的效果在立地条件较差的情况下表现得更为明显．     

马尾松与粘盖乳牛肝菌菌根共生体形成过程

对菌根共生机制的研究是对其进行应用的前提,到目前为止,绝大多数外生菌根（ectomycorrhiza,ECM）的建立过程尚不明晰,在一定程度上限制了这些ECM真菌在林业中的应用。本研究以我国南方地区主栽树种之一——马尾松Pinus massoniana和其林下优势ECM真菌——粘盖乳牛肝菌Suillus bovinus为材料,在无菌条件下研究两者菌根共生体形成过程的形态学特征。结果表明马尾松与粘盖乳牛肝菌的共生过程分为2个阶段：（1）预共生阶段,即物理接触之前,粘盖乳牛肝菌可通过释放挥发物和分泌物促进马尾松根系伸长和分枝;（2）共生阶段,又可分为3个时期。接种后第4天,粘盖乳牛肝菌菌丝体开始与马尾松根系接触并形成附着胞进入接触期;第7天菌丝开始侵入根系内部,侵入期开始;第28天菌套和哈氏网形成,即菌套和哈氏网形成期,该时期菌根化根尖开始膨大,随后继续发育至二叉分枝状菌根形成。在发育顺序方面,哈氏网与菌套同步发育,但哈氏网成形早于菌套。以上结果可对后续ECM共生机制的深入研究及马尾松高效菌根化育苗技术的开发提供参考。     

外生菌根合成过程中外源竞争性真菌及其菌根形态

在温室内,利用松属Pinus、栎属Quercus植物幼苗分别与乳菇属Lactarius、块菌属Tuber中重要食用菌进行菌根合成过程中,发现有9种外源竞争性菌根真菌exogenous competitive mycorrhizal fungi （ECMF）会造成宿主植物污染。形态解剖和分子生物学鉴定结果表明,9种ECMF分属5个属（Amphinema、须腹菌属Rhizopogon、硬皮马勃属Scleroderma、小球孢盘菌属Sphaerosporella、乳牛肝菌属Suillus）5科（阿太菌科Atheliaceae、须腹菌科Rhizopogonaceae、硬皮马勃科Sclerodermataceae、火丝菌科Pyronemataceae、乳牛肝菌科Suillaceae）,其中,造成松树苗污染的主要为Amphinema、Rhizopogon和Suillus属的种类,造成栎树苗污染的主要为Scleroderma属的种类,而Sphaerosporella属的种类既能污染松也能污染栎。同时,对ECMF形成菌根的形态与解剖特征进行了描述,以期为菌根合成过程中ECMF污染的识别与控制提供借鉴。     

红菇与红锥形成的根共生体形态的描述*

红菇与红锥的茛共生形成的菌根属外生菌根,分叉形成为单轴状,颜色呈浅黄褐色,菌根表面呈波纹状突起。菌套的外表特征呈绒毛状,厚度２６￣４０μｍ,具典型的哈蒂氏网、丰富的外延菌丝和菌索,无菌核。外延菌丝长度约４５￣１００μｍ,直径约１．８￣２．３μｍ,有分隔不分支,未风锁状联合。菌根菌的生态组合型为不规则垫簇型。     

外生菌根的形态学、解剖学及分类学研究进展

本文综述了外生菌根的形态学和解剖学特征,评价了这些特征用于外生菌根分类的价值,同时指出各种类型的形态学特征和解剖学特征与其营养吸收的关系。这些特征包括了菌根的颜色类型及变化、形状及分枝方式、外伸菌丝的多少及特征、根状菌索有无及分化、菌核有无及特征、菌套内外表面的菌丝排列及分化,菌套的切面特征,丹宁层厚度及分化、哈氏网的菌丝排列及厚度等。较详细阐述了100多年来,人们为外生菌根分类所作出的种种努力,并对各种分类方式作了简单的评价。     

离体条件下日本冷杉／二色蜡蘑的外生菌根合成

本实验报道了以离体培养方式诱导日本冷杉(AbiesfirmaSieb.etZucc.)与二色蜡蘑(Laccariabicolor(Maire)Orton)的外生菌根合成的方法。日本冷杉能否与广谱性的外生菌根菌二色蜡蘑形成人工菌根?菌根结构的诱导是否只发生在植物体整体水平上?这些是本实验的关注点。研究结果显示:无菌幼苗接种10周后,在光学显微镜下可观察到典型的外生菌根的特征,即由高度分叉的外延菌丝体形成的厚实的菌套以及由内延菌丝体形成的哈蒂氏网。将来源于下胚轴的愈伤组织与二色蜡蘑的菌丝体进行共同培养,3周后,菌丝体开始接触愈伤组织的表面,并且侵入到愈伤组织的细胞间,形成拟哈蒂氏网结构。愈伤组织可以作为一种菌根学研究的有效的培养体系。这一方法可望为冷杉类植物菌根学的研究提供有效的实验工具。     

外生菌根菌的分子生态学：分子标记基因、遗传个体及生态意义

外生菌根真菌与很多植物形成互利共生关系,在营养物质交换和碳循环等方面起着关键性的作用,是森林生态系统的重要组成部分。近期生物技术的发展使得人们对外生菌根菌的群体遗传学和分子生态学有了更加深入的认识。本文介绍了一些常用的鉴定外生菌根菌的分子标记,并对每种分子标记的特点及其适用范围进行了讨论。文中总结了几种常用的鉴定未知外生菌根菌的方法,指出了一些在研究外生菌根菌过程中需要克服的内在困难,其中之一就是很多外生菌根菌不可以人工培养,所以人们缺少对其地下部分分布规律和动态变化的了解。在寄主专一性、物种多样性和丰富度、遗传个体大小、繁殖方式等方面,近期对外生菌根菌的分子生物学研究已经获得了很多重要的结果。作者讨论了这些研究成果对于今后开展外生菌根菌研究的重要意义以及在森林生态系统保育方面的潜在应用价值。     

Comprehensive proteome analysis in Cenococcum geophilum Fr. as a tool to discover drought-related proteins

Cenococcum geophilum is a widely distributed ectomycorrhizal fungus potentially playing a significant role in resistance and resilience mechanisms of its tree hosts exposed to drought stress. In this study, we performed a large scale protein analysis in pure cultures of C. geophilum in order to gain first global insights into the proteome assembly of this fungus. Using 1-D gel electrophoresis coupled with ESI-MS/MS, we indentified 638 unique proteins. Most of these proteins were related to the metabolic and cellular processes, and the transport machinery of cells. In a second step, we examined the influence of water deprivation on the proteome of C. geophilum pure cultures at three time points of gradually imposed drought. The results indicated that 12 proteins were differentially abundant in mycelia subjected to drought compared to controls. The induced responses in C. geophilum point towards regulation of osmotic stress, maintainance of cell integrity, and counteracting increased levels of reactive oxygen species formed during water deprivation.     

Formation of Mycorrhiza-like Structures in Cultured Root/Callus of Cathaya argyrophylla Chun et Kuang Infected with the Ectomycorrhizal Fungus

An in vitro system was used for ectomycorrhizal synthesis of Cenococcum geophilum Fr. with Cathaya argyrophylla Chun et Kuang, an endangered species. Calli initiated from stem segments and adventitious roots differentiated from young seedlings were removed and cocultured with Cenococcum geophilum on a modified Murashige-Skoog medium. Fungal hyphae were visible within intercellular spaces of the callus 4 weeks after inoculation, but definite and well-developed Hartig net structures did not form in the calli 8 weeks after inoculation. The typical ectomycorrhizal structures (i.e. hyphal mantle and intracortical Hartig net) were observed in root segments 8 weeks after inoculation. This is the first report of aseptic ectomycorrhizal-like formation/infection between root organ/callus of Cathaya argyrophylla and the ectomycorrhizal fungus Cenococcum geophilum. This culture system is useful for further investigation of mycorrhizal synthesis in Cathaya trees.（Author for correspondence. Tel: +86 (0)451 8219 1783; Fax: +86 (0)451 8219 1795; E-mail: lumin-fu@163.com）     

Intracellular colonization of Rhododendron and Vaccinium roots by Cenococcum geophilum, Geomyces pannorum and Meliniomyces variabilis

Four in vitro experiments were set up to verify the colonization potential of ectomycorrhizal (EcM) Cenococcum geophilum FR. (strain CGE-4), saprotrophic Geomyces pannorum (LINK) SIGLER & CARMICHAEL (GPA-1) and a frequent root-associated, potentially ericoid mycorrhiza (ErM)-forming Meliniomyces variabilis Hambleton & Sigler (MVA-1) in roots of Rhododendron and Vaccinium. A typical ErM fungus, Rhizoscyphus ericae (Read) Zhuang & Korf (RER-1), was included for comparison. All fungal strains intracellularly colonized rooted Vaccinium microcuttings: GPA-1 occasionally produced hyphal loops similar to ErM, MVA-1 and RER-1 exhibited a typical ErM colonization pattern. CGE-4 hyphae grew vigorously on and around newly formed roots and rarely penetrated turgescent rhizodermal cells forming intracellular loose loops. Rooting of Rhododendron sp. microcuttings was not promoted by any fungal strain except CGE-4, which also promoted the most vigorous growth of Rhododendron ponticum L. seedlings. The widespread EcM fungus C. geophilum has a potential to colonize non-EcM roots and support their development which may influence overall growth of ericaceous plants. As shown for G. pannorum, structures resembling ErM may be formed by fungi that are to date not regarded as ericoid mycorrhizal.     

Mycobionts of <Emphasis Type="Italic">Salix herbacea</Emphasis> on a glacier forefront in the Austrian Alps

Dwarf willows (e.g. Salix herbacea) are among the earliest ectomycorrhizal (EM) plants colonising primary successional sites such as glacier forefronts in the Tyrolean Alps. EM of S. herbacea were sampled at the Rotmoos glacier forefront (Otz Valley, Austria) three times a year during the growing season and once a year during winter when plants were covered with snow in 2005 and 2006. EM were investigated using morphological methods and by sequencing the rDNA ITS region. The degree of EM mycorrhization was high throughout both years (93%). We distinguished 21 EM morphotypes and identified 19 fungal species. Cenococcum geophilum, Sebacina spp., Tomentella spp. and Cortinarius spp. dominated the mycobiont community of S. herbacea. The observed species richness in this about 150-year-old soil was at least 59% of the estimated species richness. Fungal communities differed significantly between consecutive years, and spatial heterogeneity was high. These differences made it difficult to detect seasonal impacts. Abundances of C. geophilum EM increased throughout the 2-year sampling period. Sebacina incrustans EM were very abundant in 2005, but nearly disappeared in 2006, whilst its fruitbodies were still frequent in the sampling area. This suggests that the mycorrhizae were displaced from the roots by an outcompeting species, whereas the mycelium was still present in the soil.     

Significant diversity and potential problems associated with inferring population structure within the Cenococcum geophilum species complex

Cenococcum geophilum is perhaps the most widely distributed and most recognized ectomycorrhizal fungus with a host range of more than 200 tree species from 40 genera of both angiosperms and gymnosperms. We conducted a phylogenetic analysis on a large collection of isolates (n=74) from North America and Europe based on glyceraldehyde 3-phosphate dehydrogenase (gpd). A subset of isolates (n=22) also was analyzed with the more conservative LSU-rDNA locus. Significant nucleotide diversity was detected (approximately 20%) in the gpd region and the LSU-rDNA analysis supported that the C. geophilum isolates studied were monophyletic but distinct from two isolates, Am5-1 and N2-10, which previously were used in population genetic studies of this species. These results suggest that Am5-1 and N2-10 are likely two undescribed species or even genera. Our results suggest that C. geophilum sensu lato is a species complex and support previous molecular, physiological and morphological studies that have shown significant diversity in C. geophilum. This study also revealed that caution is advised when conducting population genetic studies in C. geophilum due to the possibility of pooling unrelated isolates. This potential problem also has implications for other fungal taxa because cryptic species routinely have been found in recent years based on molecular data.     

Effect of nitrate and ammonium concentration on nitrate reductase activity in five species of mycorrhizal fungi

The nitrate reducing capacity of pure cultures of Cenococcum geophilum (Sow.) Ferd. & Winge, Paxillus involutes (Batsch: Fr.) Fr. (strains 1 and 2), Piloderma croceum Erikss. & Hjortst., Suillus variegatus (Fr.) O. Kuntze (strains 1 and 2) and an ectendomycorrhizal (E-strain) fungus was measured using an in vivo nitrate reductase (EC 1.6.6.3) assay. Differences between species and strains were established. The nitrate concentration of the culture medium influenced the nitrate reductase activities of the E-strain fungus and one strain of S. variegatus. The nitrate reductase activity of certain species and strains was a function of nitrate concentration. Addition of ammonium to the growth medium did not have any significant effect on the in vivo or in vitro nitrate reductase activity. The in vivo nitrate reductase activity in the mycelia of C. geophilum and the E-strain fungus decreased during 28 day growth in modified Melin-Norkrans medium. For mycelia of Paxillus involutus, Piloderma croceum and S. variegatus grown on agar the in vitro assays showed higher nitrate reductase activity than the in vivo assays.     

Effects of Mineral Structure and Microenvironment on K Release from Potassium Aluminosilicate Minerals by Cenococcum geophilum fr

K-bearing minerals with enormous reserve in the world, were not easy to provide the available K nutrient element for plant direct uptake because of the slow K release rate. With the potential potassium-solubilizing microorganisms, the slow K release rate from minerals would be improved significantly. In this work, Cenococcum geophilum Fr, one of the most common ectomycorrhizal fungi in boreal to temperate regions, was adopted to dissolve K-bearing minerals for K release. Five kinds of potassium aluminosilicate minerals were tested by bioleaching experiments in pure culture, including feldspar, nepheline, biotite, muscovite and illite. The available and unavailable potassium amounts in minerals before and after bioleaching were measured and compared with each other. The effect of mineral structure on the potassium solubilization efficiency by Cenococcum geophilum Fr. was discussed. Furthermore, the microenvironment formation between fungi and mineral surface to enhance the K release rate was investigated through detecting K, Al, Si concentrations and metabolites amounts (polysaccharide and organic acids) in microenvironment and external environment, respectively. Experimental results demonstrated that Cenococcum geophilum Fr. was a potential candidate of potassium solubilizing microorganisms, and both mineral structure and microenvironment have significant effects on the K release rate.     

土生空团菌的遗传多样性分析

为了研究土生空团菌的遗传多样性,对来自中国、美国、瑞士和法国等菌株的rDNA ITS区进行序列分析,利用Popgene32和phylip软件进行数据计算和聚类分析。序列分析表明土生空团菌rDNA ITS区的序列长度为422–447bp,遗传距离在0.000–0.051之间。居群结构和聚类分析结果表明:(1)土生空团菌有一定的遗传多样性,且遗传差异主要来自于居群内;(2)基因流Nm>1,遗传漂变不是导致土生空团菌居群遗传分化的主要因素;(3)土生空团菌的遗传分化受到地理环境的影响,而与宿主来源没有明显相关性。     

Formation of Mycorrhiza-like Structures in Cultured Root/Callus of Cathaya argyrophylla Chun et Kuang Infected with the Ectomycorrhizal Fungus Cenococcum geophilum Fr.

An in vitro system was used for ectomycorrhizal synthesis of Cenococcum geophilum Fr. with Cathaya argyrophylla Chun et Kuang, an endangered species. Calli initiated from stem segments and adventitious roots differentiated from young seedlings were removed and cocultured with Cenococcum geophllum on a modified Murashlge-Skoog medium. Fungal hyphae were visible within intercellular spaces of the callus 4 weeks after inoculation, but definite and well-developed Hartig net structures did not form in the calU 8 weeks after inoculation. The typical ectomycorrhizal structures （i.e. hyphal mantle and Intracortical Hartig net） were observed in root segments 8 weeks after inoculation. This is the first report of aseptic ectomycorrhlzal-like formation/infection between root organ/callus of Cathaya argyrophylla and the ectomycorrhizal fungus Cenococcum geophflum. This culture system is useful for further investigation of mycorrhizal synthesis in Cathaya trees.     

辽东栎幼苗的外生菌根合成

 在温室花盆中播种辽东栎（Quercus liaotungensis）种子获得辽东栎幼苗,并对幼苗接种外生菌根真菌进行菌根合成试验。所用的外生菌根真菌有：铆钉菇（Gomphidius viscidus）、臭红菇（Russula foetens）、厚环乳牛肝菌（Suillus grevillei）、褐环乳牛肝菌（S. luteus）、彩色豆马勃（Pisolithus tinctorius）、美味牛肝菌（Boletus edulis）、赭丝膜菌（Cortinarius russus）、土生空团菌（Cenococ