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。     

川西亚高山岷江冷杉外生菌根形态随海拔梯度的分化

外生菌根真菌是一类有助于植物生长及应对环境变化的重要共生真菌类群。深入认识外生菌根特征有助于理解植物对环境变化的响应规律。本研究在四川省卧龙自然保护区巴郎山脉设置海拔2850、3000、3194、3413和3593 m 5个样地,利用中心象限法进行土块样品(10 cm×10 cm×10 cm)采集,通过分析各海拔每土块中岷江冷杉外生菌根形态个数、形态多样性及养分获取类型等指标,研究岷江冷杉外生菌根形态特性随海拔的变异特征及土壤环境因子的作用。结果表明: 1) 菌根形态为橘黄色或橘色、单轴羽状分支或不分支、菌套表面光滑、无或少见外延物、根尖直筒型或膨大的菌根类型是卧龙自然保护区岷江冷杉外生菌根的主要形态类型。该形态类型在研究区内的侵染率最高,为12.4%;2) 随着海拔的升高,岷江冷杉外生菌根形态多样性下降,每土块内的形态个数减少;3) 单因素方差分析结果表明,不同海拔上无外延菌丝或极少外延菌丝的紧密接触类型(CE)和短距离养分获取类型(SDE)外生菌根侵染率有显著差异,CE类型外生菌根侵染率随着海拔的升高而增加;4) 土壤环境因子对岷江冷杉外生菌根形态在海拔上的变异有一定的影响,冗余分析(RDA)表明,土壤全氮含量、全磷含量、土壤温度、土壤含水率、pH值及酸性磷酸酶活性对岷江冷杉外生菌根形态在海拔上的变异有显著影响,其中土壤全氮含量和土壤温度的影响最大,分别解释了岷江冷杉形态特征变异的5.4%和4.9%。本研究揭示了岷江冷杉外生菌根形态形成在海拔上的差异,为进一步研究亚高山针叶树种菌根策略对环境变化的响应机制提供了科学依据。     

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。     

七株外生菌根真菌与三种松苗菌根的形成能力

松树外生菌根菌资源丰富,但实际应用的种类不多。为筛选出与松苗形成菌根能力较强的菌种,采用播种接菌和芽苗截根移栽接菌两种方法,对7株外生菌根真菌与马尾松、湿地松和黑松3种松苗的菌根合成进行了研究。结果表明：形成的菌根以二叉分枝状为主,棒状菌根相对较少,多叉状菌根以马尾松较多。PC2形成的菌根表面菌丝厚且紧密,504、EG、Pt,形成的菌根表面菌丝紧密程度中等,而505、ZJ和HX形成的菌根其表面菌丝则比较稀疏;Pt1、Pt2、EG形成的菌根外延菌丝较长,而505、HX形成的菌根外延菌丝极短。截根接菌时的感染率和感染指数要高于播种接菌。504形成菌根的能力最强,在3种松苗上的菌根感染率都达100％,感染指数最高可达90,最低也达70;Pt2和EG与马尾松和黑松形成菌根的能力较强;而505和HX仅与马尾松形成菌根的能力较强;Pt1形成菌根的能力较差,在3种松苗上菌根感染率和感染指数都较低。在3种松苗中,马尾松的菌根化状况最好,其次为黑松,湿地松的菌根化状况较差。     

外生菌根与植物抗重金属胁迫机理

外生菌根是外生菌根真菌和植物营养根形成的共生体,能够增加植物对污染胁迫的抵抗能力。本文综述了20多年来国内外研究外生菌根增加植物抗重金属毒害的成果,指出了外生菌根在植物抗重金属毒害中的积极作用,并概括其抗性的主要机理为：外延菌丝的吸收作用;菌根分泌物的调节与螯合作用;菌根菌套或哈蒂氏网吸收过滤有毒金属;菌根菌套的疏水性作用。在研究外生菌根抗重金属毒害机理的基础上,提出了该领域今后的研究前景。     

中国叶生地衣型真菌2新种和1新记录种

基于形态、解剖、化学等研究方法,报道了中国的地衣型真菌2个新种和1个新记录种,分别是黄绿浅盘衣Calenia flavescens、多疣浅盘衣Calenia verrucosa和黑盘刺衣Echinoplaca fusconitida。黄绿浅盘衣的主要特征是地衣体浅黄绿色,具不明显的白色疣体,盘面黄色,子囊含2或4个孢子,子囊孢子(2～) 3～7隔,15. 5～22.5×4.0～8.5μm,分生孢子柱刚毛状,长约0.15 mm,白色。多疣浅盘衣的主要特征是地衣体浅橄榄绿色,具与地衣体同色且密集的疣体,盘面褐色,子囊含(6～) 8个孢子,子囊孢子(2～) 3～5隔,15.5～25.0×6.5～8.5μm,分生孢子柱刚毛状,长0.4～1.5 mm,白色至浅黄色。黑盘刺衣的地衣体为壳状,具白色疣状突起和黄白色刚毛,子囊盘直径0.2～0.5 mm;盘面平坦,灰褐色至黑褐色,子囊含1个孢子,子囊孢子无色,砖壁型,35～65×15～20μm。文中详细描述了3个种的形态特征并提供了图片。     

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.     

两种乳菇菌丝生长的最适培养基与菌根合成

通过测量菌丝直径和干重对松乳菇（rcb-74）和靓丽乳菇（rll-107,rmsh-118）进行最适菌丝生长培养基的筛选。结果表明所有菌株均在改良的biotin-aneurine-folic acid（BAF）培养基上表现出较大的菌丝直径以及最大菌丝干重。用改良的BAF培养基制备乳菇液体菌丝,接种云南松、马尾松和油松,并在13-30d后发现rcb-74与云南松和油松形成菌根,rll-107与马尾松形成菌根,rmsh-118与云南松形成菌根。     

块菌发育过程中伴生微生物多样性及其生态功能

块菌属Tuber是子囊菌门中一类珍贵的地下生外生菌根真菌,能与松科、壳斗科等多种树木形成菌根共生关系,在森林生态系统中扮演着重要的角色。同时,该属的一些种类因其独特的香味而备受国际食用菌市场推崇,具有极高的经济价值。在块菌属漫长的生活史中,菌丝的生长、菌根的形成、子囊果的产生和发育、特殊芳香化合物及活性物质的产生和积累均与其伴生微生物有着密切的关系。近年来,分子生物学技术的发展尤其是高通量测序技术的应用推动了对块菌属子囊果形成和发育机制的研究,为块菌生态学研究提供了有力的工具,使块菌伴生微生物相关研究取得重要突破。本文综述了近5年来国内外对块菌属生长发育过程中伴生细菌、真菌多样性及其生态功能的相关研究,并探讨了其中尚未清楚的问题及今后可能的研究方向,为块菌属生物学的研究及人工栽培奠定基础。     

块菌人工栽培现状及菌根苗培育方法的研究进展

松露(块菌)是名贵的地下真菌,因其独特的香味和口感享誉全球,具有重要的经济价值、食药用价值和生态价值。块菌必须与宿主植物共生形成外生菌根后才能形成子实体(子囊果)。由于块菌具有较高的经济价值而野生块菌资源被严重破坏,以块菌菌根苗制备技术为基础的人工栽培手段受到广泛关注。本文以块菌菌根苗培育为切入点,阐述了块菌资源面临的较大威胁、人工栽培手段的进步和人工种植园的快速发展,通过综述菌根苗培育前对共生组合的选择,菌根苗培育时使用的基质、接种剂、培养方式、与其他生物间的互作,以及菌根苗培育后进行检测的方法等研究进展,为外生菌根食用菌的菌根苗培育技术和人工栽培推广提供参考。     

Mycorrhization of Pecan trees (Carya illinoinensis) with commercial truffle species: Tuber aestivum Vittad. and Tuber borchii Vittad

Pecan (Carya illinoinensis) is an economically important nut tree native to the Mississippi basin and cultivated worldwide. In North America, species of truffles are regularly found fruiting in productive pecan orchards and the truffle genus Tuber appears to be abundant in pecan ectomycorrhizal (EM) communities. As an initial step to determine the feasibility of co-cropping European truffle species with pecan, we evaluated whether mycorrhizae of highly esteemed European truffle species (Tuber aestivum Vittad. T. borchii and T. macrosporum) could be formed on pecan seedlings. Seedlings were inoculated with truffle spores and were grown in a greenhouse for 10?months. Levels of EM colonization were estimated visually and quantified by counting EM tips. Ectomycorrhizae were identified both morphologically and molecularly with species-specific amplification and by sequencing of the ITS region of the nuclear ribosomal DNA (nrDNA). Both T. borchii and T. aestivum spores produced well-formed ectomycorrhizae on pecan seedlings with average root colonization levels of about 62% and 42%, respectively, whereas no ectomycorrhizae of T. macrosporum were formed. The anatomy and morphology of these truffle ectomycorrhizae on pecan was characterized. The co-cropping of T. aestivum and T. borchii may hold promise as an additional stream of revenue to pecan growers, although, further studies are needed to assess whether this symbiosis is maintained after planting in the field and whether truffle production can be supported by this host species.     

云南松幼苗棉革菌属菌根真菌的物种多样性

以滇中1～2年生云南松幼苗为研究对象,观察鉴定与其共生的棉革菌属真菌外生菌根真菌.通过形态观察发现了6种形态型菌根.文中对这6种形态型菌根的外观和显微特征进行了详细描述.对rDNAITS片段比对分析结果表明,6种形态型对应6种棉革菌属真菌,它们分别是与Tomentella badia、T.coerelea、T.viri...     

Species Diversity of Russuloid Mycorrhizae forming Fungi on Pinus yunnanensis Seedlings and the Mycorrhizal Morphology

Russuloid ectomycorrhizae on 1-2 years old seedlings of Pinus yunnanensis collected from the central Yunnan, China were investigated. Six morphotypes were recognized by macro and anatomical morphological approaches as well as molecular analyses. The six morphotypes were confirmed to represent six phylotypes by matching rDNA ITS sequences. The fungal partners in the six morpho phylotypes are those closely related with Russula livescens, Rvioleipes ,Rdensifolia ,Rnigricans ,Rsanguinea and Rnauseosa respectively. The correlation between the morphology of mantles and cystidia and the phylogenetic clades is further supported in our work. The framework, in which this report is included, shows that russuloid mycorrhizae are one of the most dominant representative ectomycorrhizae formed on 1-2 years seedlings of Pinus yunnanensis.     

Species-specific primers for the identification of the ectomycorrhizal fungus Tuber macrosporum Vittad

Limitations in the use of morphological traits to identify ectomycorrhizae have led to the development of species‐specific molecular markers. Herein, we report a PCR‐based technique for the reliable molecular identification of the ectomycorrhizal fungus Tuber macrosporum Vittad. Species‐specific primers were designed from an alignment of internal transcribed spacer rDNA sequences from Tuber spp. and from the most common ectomycorrhizal contaminants found in the root systems of truffle‐infected plants. The primers were tested for selective amplification using both different truffles and different ectomycorrhizae and were found to identify T. macrosporum successfully. The application of the primers in certifying the quality of truffle‐inoculated seedlings is discussed.     

Molecular identification of Tuber magnatum ectomycorrhizae in the field

Tuber ectomycorrhizae in a Tuber magnatum "truffière", located in Central Italy, were studied using molecular methods. Specifically, RFLP-ITS analyses, ITS sequencing and specific probes hybridization were used to identify 335 Tuber-like ectomycorrhizal morphotypes. Molecular identification was possible even when distinct morphological characteristics were lacking. For the first time, T. magnatum ectomycorrhizae and other coexisting Tuber species collected in the field were analysed using molecular tools for unambiguous identification. Although the "truffière" under investigation yields good harvests of T. magnatum fruiting bodies, the percentage of T. magnatum ectomycorrhizae found was very low (less than 4.4% of the 335 root tips analysed), whereas the percentages of Tuber maculatum and Tuber rufum were considerably higher (48.9% and 19.0%, respectively).     

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。     

云南松幼苗上红菇类菌根真菌的物种多样性及其菌根形态

以滇中1～2年生云南松幼苗为研究对象,观察鉴定与其共生的红菇属真菌外生菌根。形态观察发现了6种形态型（morphotypes）。本文对这6种形态型的外观和显微特征进行了详细描述,尤其强调了菌套形态特征。对rDNAITS片段比对分析表明,6种形态型对应6种红菇属真菌,它们分别是与Russula li-vescens、R.violeipes、R.densifolia、R.nigricans、R.sanguinea及R.nauseosa相近的红菇种类。本研究表明,形成的菌根及其菌套和囊状体的特征在红菇属真菌的系统分支间表现出较为稳定的差异。这一研究与前人对该属真菌的菌根形态及分类学研究基本吻合。红菇属真菌是云南松1～2年幼苗期的主要共生真菌类群。     

Biochemical, electrophoretic and immunohistochemical aspects of malate dehydrogenase in truffles (Ascomycotina)

The malate dehydrogenase (MDH; EC 1.1.1.37; L-malate-NAD(+)-oxidoreductase) activities of truffles of the genus Tuber (Tuber melanosporum Vittad., Tuber brumale Vittad., Tuber aestivum Vittad., Tuber magnatum Pico, Tuber rufum Pico) have been characterized with regard to the K(m) and V(max) values in the direct and reverse reactions. The isoelectrofocusing has revealed bands showing pI values ranging from pH 5.85 to 7.8. The MDH of T. melanosporum has been partially purified by hydroxyapatite treatment, DEAE-cellulose and Sephadex G-75 columns. With the partially purified T. melanosporum MDH activity polyclonal anti-T. melanosporum MDH antibodies have been prepared and used to localize MDH in the mycorrhizae and ascocarps of T. melanosporum. These antibodies inhibit T. melanosporum MDH activity as well as that of T. magnatum but not that of rabbit liver; this supports the specificity of the MDH antibodies used to localize MDH in truffle tissues.