New data on ectomycorrhizae and soils of the Chinese truffles Tuber pseudoexcavatum and Tuber indicum, and their impact on truffle cultivation

Chinese truffles serve as a good complement to the market for Tuber melanosporum (Périgord black truffle). However, Chinese truffles could be introduced accidentally or fraudulently into the plantations of Mediterranean truffles, and they could have a negative effect on truffle production and natural ecosystems. The study of Tuber species from China which are commercialized in Europe began 14 years ago. Tuber pseudoexcavatum was proposed as a new species, and this has been validated by some authors based on molecular and phylogenetic studies. We synthesize their ectomycorrhizae using samples from the type collection, and we compare T. pseudoexcavatum and Tuber indicum ectomycorrhizae. The ectomycorrhizae of these species have a morphology which is related to the ectomycorrhizae of T. melanosporum. We provide useful information for the rapid screening of the above-mentioned Chinese truffles ectomycorrhizae, for the quality control of commercial plants mycorrhized with Tuber. Moreover, we analyze the soil tolerance and the host plant affinity of T. pseudoexcavatum and T. indicum, in order to assess the capacity of both Chinese truffles to penetrate T. melanosporum plantations and habitats.     

Nitrogen fixation associated with Suillus tomentosus tuberculate ectomycorrhizae on Pinus contorta var. latifolia

BACKGROUND AND AIMS: Tuberculate ectomycorrhizae are a unique form of ectomycorrhiza where densely packed clusters of mycorrhizal root tips are enveloped by a thick hyphal sheath to form a tubercle. The functional significance of such a unique structure has not previously been established. The purpose of the present study was to investigate and measure the potential nitrogenase activity associated with Suillus tomentosus/Pinus contorta tuberculate ectomycorrhizae in two stand ages, young and old, and across a range of nitrogen-poor soil conditions. METHODS: Short roots were compared with other mycorrhizae and non-mycorrhizal secondary roots using tuberculate ectomycorrhizae. Assessment of nitrogenase activity was determined and quantitative measurements were taken on tuberculate ectomycorrhizae in situ in a variety of different circumstances, by using an adaptation of the acetylene reduction assay. KEY RESULTS: Significant nitrogenase activity was measured associated with S. tomentosus/P. contorta tuberculate ectomycorrhizae whereas no nitrogenase activity was measured with non-tuberculate mycorrhizae or secondary roots without mycorrhizae. Average nitrogenase activity ranged from undetectable to 5696.7 nmol C2H4 g(-1) tubercle 24 h(-1). Maximum nitrogenase activity was 25,098.8 nmol C2H4 g(-1) tubercle 24 h(-1). Nitrogenase activity was significantly higher in young stands than in old stands of P. contorta. Season or some covariate also seemed to affect nitrogenase activity and there was suggestion of a site effect. CONCLUSIONS: Suillus tomentosus/P. contorta tuberculate ectomycorrhizae are sites of significant nitrogenase activity. The nitrogenase activity measured could be an important contribution to the nitrogen budget of P. contorta stands. Season and stand age affect levels of nitrogenase activity.     

Ectomycorrhizae of Tomentella albomarginata (Thelephoraceae) on Scots pine

 The ectomycorrhizae of Tomentella albomarginata are comprehensively described and compared to ectomycorrhizae of other Tomentella species and to ectomycorrhizae of some members of Thelephoraceae ss. Stalpers and Bankeraceae ss. Stalpers. The ectomycorrhizae of T. albomarginata are characterized by a hyphal net lying on a pseudoparenchymatous mantle surface, by tubular outgrowths of irregularly angular mantle cells, and by clamp-bearing emanating hyphae. Accepted: 15 July 1995     

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).     

Exploration types of ectomycorrhizae

There is a need to quantify and qualify the ability of ectomycorrhizae to improve tree growth and nutrition and, in particular, to define criteria to classify ectomycorrhizae with respect to ecologically relevant features. Whereas the numbers of ectomycorrhizae and morphotypes give useful information when related to root biomass, root length or soil volume, the development and differentiation of the extramatrical mycelium may represent important predicative features relevant to the ecological classification of ectomycorrhizae. Here, different exploration types of ectomycorrhizae are distinguished based on the amount of emanating hyphae or the presence and differentiation of rhizomorphs. Their putative ecological importance is discussed. Accepted: 27 March 2001     

Ectomycorrhizal syntheses with Picea abies and three fungal species: a case study on the use of an in vitro technique to identify naturally occurring ectomycorrhizae

Ectomycorrhizal syntheses between Picea abies and the fungal associates Scleroderma citrinum, Boletus luridus, and Tricholoma vaccinum were carried out using Melin's Erlenmeyer flask technique. The symbioses of S. citrinum were characterized by a mantle composed of an outer prosenchymatous and an inner synenchymatous layer. The mantles of B. luridus and T. vaccinum were solely prosenchymatous. Rhizomorphs were produced in all treatments, but only in association with S. citrinum were they differentiated with additional, enlarged hyphae. All synthesized ectomycorrhizae were white or whitish to light orange and greyishorange. On large-scale root sampling in two differing Picea abies forests in Switzerland, nine out of a total of 22 morphological types of ectomycorrhizae were white or yellow in colour and were, therefore, comparable with the synthesized ectomycorrhizae. These nine natural types generally had distinct mantle features (irregular synenchyma, gelatinous matrix, cystides, thick-walled hyphae), but mostly lacked clamp connections. Synthesized ectomycorrhizae, on the other hand, lacked distinct mantle characteristics and always had clamp connections. Natural and synthesized white or yellow ectomycorrhizae did not coincide morphologically and thus identification of the fungal partners of natural symbioses by means of in vitro-synthesis with potential ectomycorrhizal fungi was not possible in the present study.     

The Ecological Relation Between Pinus taiwanensis and Fungi in the Process of Ectomycorrhizae Synthesis

The root samples of Pinus taiwanensis from seedlings to large trees of Mt. Huangsban were studied under light microscope(LM) and scanning electron microscope(SEN). It is obvious that ectomycorrhizae from extensively between the pine roots and the fungi. Isolation and pure culture of the samples of sporocarps were carried out, of which six strains were inoculated to the dissinfected seedlings were under laboratory condition. Pisolithus tinctorius was introduced to inoculate the pine seedlings too. Studies on the development of dichotomous mycorrhizae and the early stage of ectomycorrhizae formation were specially emphasized. Three types of the process of hyphae infection and the ontogeny of ectomycorrhizae synthesis, development and subsequent degeneration were described. Some ecological effects of ectomycorrhizae, which increase the growth of pine trees were also mentioned. Finally the authors sugge. st that the success inoculation of Pisolithus tinctorius to the pine trees will offer a new device for the formation and development of the ectomycorrhizae in terms to promote their growth.     

Ectomycorrhizae between <Emphasis Type="Italic">Alnus acuminata</Emphasis> H.B.K. and <Emphasis Type="Italic">Naucoria escharoides</Emphasis> (Fr.:Fr.) Kummer from Argentina

Field ectomycorrhizae of Naucoria escharoides on Alnus acuminata ("andean alder", "aliso del cerro") are described in detail for the first time. Naturally occurring ectomycorrhizal roots were sampled beneath sporocarps of N. escharoides. The samples were taken from four natural forest plots at two homogeneous A. acuminata sites (Tucumán and Catamarca Provinces, Argentina). The ectomycorrhizae were characterized morphologically and compared by means of PCR/RFLP analysis of the internal transcribed spacer region of the nuclear rDNA. The most important morphological features of the ectomycorrhizae are a white to pale yellow mantle, simple to monopodial branches, hyaline emanating hyphae, abundant hyphal bundles emerging more or less perpendicularly from a plectenchymatous mantle, and an acute or rounded apex with or without a mantle. N. escharoides fruitbodies have white basal mycelium with emanating hyphae similar to those of andean alder ectomycorrhizae. The RFLP profiles of sporocarps and mycorrhizae were the same.     

Ectomycorrhizae of Sarcodon imbricatus on Norway spruce and their chlamydospores

Summary The ectomycorrhizae of Sarcodon imbricatus are comprehensively described and compared to other ectomycorrhizae of Thelephoraceae species. Sarcodon imbricatus ectomycorrhizae are distinguished from all previously described ectomycorrhizae by the unique shape of their chlamydospores.Considered as part XXXVI of the series Studies on ectomycorrhizae; part XXXV: Weiss (1991)     

Ectomycorrhizae of Phellodon niger on Norway spruce and their chlamydospores

The ectomycorrhizae of Phellodon niger are comprehensively described and compared to other ectomycorrhizae of Thelephoraceae species. P. niger ectomycorrhizae are distinguished from all previously described ectomycorrhizae by the unique shape of their chlamydospores.Considered as part XLIII of the series Studies on ectomycorrhizae; part XLII: Raidl and Agerer (1992)     

夏块菌与青刚栎形成外生菌根形态变化的研究

夏块菌（Tuber aestivum）是一种具有较高经济价值的菌根食用菌。对夏块菌与青刚栎（Cycloba-lanopsis glauca）在形成菌根过程中不同阶段的菌根形态变化进行了研究,结果显示：用夏块菌孢子液接种青刚栎苗后,第14天起开始形成淡乳色的外生菌根,外延菌丝刚毛状;第一至第二个月可形成黄褐色、褐色外生菌根,外延菌丝刚毛或羊毛状。外生菌根为单根,长1～4mm,直径150～250μm。菌套厚12～20μm,平坦或自菌根延伸出刚毛状菌丝,外延菌丝束黄绿色;哈替氏网菌丝直径1～1.5μm。菌根老化后变暗褐或萎缩。外延菌丝束呈黄绿色是夏块菌菌根区别于其它块菌菌根最重要的形态特征。     

Ectomycorrhizae of Lactarius lignyotus on Norway spruce,characterized by anatomical and molecular tools

The ectomycorrhizae of Lactarius lignyotus on Norway spruce are comprehensively described by morphological and anatomical characteristics. Identification of ectomycorrhizae was performed by tracing mycelia to the fruitbodies and also by molecular tools, using polymerase chain reaction (PCR) amplification of the fungal DNA. The newly described ectomycorrhiza is compared to ectomycorrhiza of the related Lactarius picinus. The amplified DNA products of the two fungi and their ectomycorrhizae could be distinguished by characteristic fragments after digestion with Hinf1.Considered as part LV of the series Studies on ectomycorrhizae of the Institute for Systematic Botany, Munich; part LIV: Agerer et al. (1994)     

A rapid mini-prep method for isolation of ectomycorrhizal DNA from Tuber species

A rapid procedure has been developed to isolate DNA from the ectomycorrhizae of Tuber spp. for use in PCR experiments. The method described is fast and sensitive and can overcome the amplification problems that can arise in the presence of inhibitors. For this reason it can be used to type ectomycorrhizae even starting from a single root tip and make mycorrhizae identification much more rapid.     

广西凭祥红锥-马尾松混交林菌根际微生物群落结构

以广西凭祥红锥-马尾松混交林中红锥和马尾松为研究对象,采集林下外生菌根和根际土壤,利用高通量测序分析该混交林下的菌根际微生物群落结构.结果 表明,红锥、马尾松菌根际优势真菌为红菇属Russula、被孢霉属Mortierella、乳菇属Lactarius、鹅膏属Amanita、拟锁瑚菌属Clavulinopsis、丝盖伞...     

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.     

Fossil ectomycorrhizae from the Middle Eocene

Fossil ectomycorrhizae were found recently among permineralized plant remains in the middle Eocene Princeton chert of British Columbia. The ectomycorrhizae are associated with roots of Pinus and have a Hartig net that extends to the endodermis, a pseudoparenchymatous mantle, and contiguous extramatrical hyphae that are simple-septate. The mycorrhizal rootlets lack root hairs and dichotomize repeatedly to form large, coralloid clusters. Reproductive structures are absent. Based on the morphological characteristics, and the identity of the host, the closely related basidiomycete genera Rhizopogon and Suillus are suggested as comparable extant mycorrhizal fungi. These exquisitely preserved specimens represent the first unequivocal occurrence of fossil ectomycorrhizae and demonstrate that such associations were well-established at least 50 million years ago.     

Index of unidentified ectomycorrhizae

 This latest instalment of the index of unidentified ectomycorrhizae lists binomials and reference specimen characterizations of ectomycorrhizae first described in 1995–1997. Accepted: 14 May 1998     

The extraction and quantification of ergosterol from ectomycorrhizal fungi and roots

Recently, ergosterol analysis has been used to quantify viable fungal biomass in resynthesized ectomycorrhizae. An objective of our study was to quantify ergosterol in a range of ectomycorrhizal isolates under differing growth conditions. In addition, we tested the applicability of the method on field-collected roots of ectomycorrhizal and vesicular-arbuscular (VA) mycorrhizal plants. Quantification of sitosterol as a biomass indicator of plant roots was also undertaken. Ergosterol was not detected in roots of uninoculated Betula populifolia seedlings, and sitosterol was not detected in an ectomycorrhizal fungal isolate but was present in birch roots. Ergosterol was produced in all isolates examined, which represented the major orders of ectomycorrhizal fungi. The range of values obtained, from 3 to nearly 18 g ergosterol mg^-1 dry mass, agrees well with reported values for other mycorrhizal and decomposer fungi. Hyphal ergosterol was the same during growth on phytic acid and KH₂PO₄. Reduction of growth temperature from 25° C to 15° C had little effect on ergosterol content of cultures harvested at similar growth stages. Ergosterol and sitosterol were detected in field-collected ectomycorrhizae of B. populifolia and Pinus sylvestris and VA mycorrhizae of Acer rubrum and Plantago major. Both ergosterol content and ergosterol to sitosterol ratios were significantly lower in VA mycorrhizae than ectomycorrhizae. Calculations of viable fungal biomass associated with field-collected roots were in agreement with those reported by others using the method on resynthesized ectomycorrhizae. Estimates of total mass could be obtained for field-collected B. populifolia roots by a simultaneously using ergosterol to estimate fungal biomass and sitosterol to estimate root mass. Some potential applications and limitations of sterol quantification in studies of mycorrhizal physiology and ecology are discussed.