Tropical Monodominance: A Preliminary Test of the Ectomycorrhizal Hypothesis1

This study reports results from the first explicit test of the ectomycorrhizal hypothesis for tropical monodominance in the Ituri Forest of the Democratic Republic of Congo (formerly Zaire), where the canopy tree Gilbertiodendron dewevrei forms large, monospecific stands. To test the hypothesis that ectomycorrhizae are important to the success of dominant species, we surveyed the mycorrhizal status of dominant species, as well as other common, but not dominant, species in the forest. The survey reveals that two dominant species, Gilbertiodendron dewevrei and Julbernardia seretii, form ectomycorrhizae and vesicular-arbuscular mycorrhizae, while Cynometra alexandri, another dominant, forms only vesicular-arbuscular mycorrhizae. These results, along with those of other species in this and other forests, are discussed within the context of the ectomycorrhizal hypothesis for tropical mondominance. This study demonstrates that the relationship between EM and tropical monodominance is more complex than has been previously recognized.     

连续三年夜间增温和施氮对云杉外生菌根及菌根真菌多样性的影响

以西南亚高山针叶林建群种粗枝云杉(Picea asperata)为研究对象,采用红外加热模拟增温结合外施氮肥(NH₄NO₃ 25 g N m^-2 a^-1)的方法,研究连续3a夜间增温和施肥对云杉幼苗外生菌根侵染率、土壤外生菌根真菌生物量及其群落多样性的影响。结果表明:夜间增温对云杉外生菌根侵染率的影响具有季节性及根级差异。夜间增温对春季(2011年5月)云杉1级根,夏季(2011年7月)和秋季(2010年10月)云杉2级根侵染率影响显著。除2011年7月1级根外,施氮对云杉1、2级根侵染率无显著影响。夜间增温对土壤中外生菌根真菌的生物量和群落多样性无显著影响,施氮及增温与施氮联合处理使土壤中外生菌根真菌生物量显著降低,但却提高了外生菌根真菌群落的多样性。这说明云杉幼苗外生菌根侵染率对温度较敏感,土壤外生菌根真菌生物量及其群落多样性对施氮较敏感。这为进一步研究该区域亚高山针叶林地下过程对全球气候变化的响应机制提供了科学依据。     

Establishment of ectomycorrhizae on the roots of two species of Eucalyptus from fungal spores contained in the faeces of the long-nosed potoroo (Potorous tridactylus)

Abstract The long-nosed potoroo (Potorous tridactylus), a medium-sized, ground-dwelling marsupial, inhabits the foothills and coastal forests of southeastern mainland Australia. Faecal analysis confirmed the mycophagous habit of P. tridactylus at a eucalypt forest site in East Gippsland, Victoria; in one month, the spores of 33 different fungi were identified. The majority of species (27) were hypogeal or subhypogeal gasteromycetes that form ectomycorrhizae with forest trees and shrubs, and included the widespread and common Mesophellia pachytrix. The possible role of mycophagy in establishment of eucalypt-fungal mycorrhizae was investigated in several ways. Fungal spores were viable after passage through the gut of P. tridactylus; application of spore-bearing faeces to seedlings of Eucalyptus sieberi and Eucalyptus globoidea grown in sterile soil produced ectomycorrhizae. Spores of M. pachythrix were among the most common spores in these faeces and some of the resulting ectomycorrhizae were of the type expected from M. pachythrix. In contrast, direct application of M. pachythrix spores from sporocarp tissue to seedlings produced no ectomycorrhizae. Ectomycorrhizae also were established on seedlings grown in unsterilized forest soil, indicating that soil-borne spores had been conditioned, either by passage through the gut of a mammal or some other means, for reaction with the roots of eucalypts. Addition of faeces improved root and shoot growth of seedlings, although we could not determine whether this was primarily due to the formation of ectomycorrhizae, the addition of nutrients in the faeces, or a combination of both. Potorous tridactylus may enhance the re-establishment of mycorrhizal colonies in logged and/or burned forest sites by dispersing in its faeces, the spores of mycorrhizal fungi from sporocarps consumed within the disturbed area, as well as spores from sporocarps eaten in adjacent undisturbed habitat. In performing this function, P. tridactylus may also aid in the recovery of regenerating silvertop ash-stringybark forests.     

The early-stage ectomycorrhizal Thelephoroid fungal sp. is competitive and effective on<Emphasis Type="Italic"> Afzelia africana</Emphasis> Sm. in nursery conditions in Senegal

This study was conducted to evaluate the competitiveness and effectiveness of Thelephoroid fungal sp. ORS.XM002 against native ectomycorrhizal fungal species colonizing potted Afzelia africana seedlings during 3 months of growth in different forest soils collected from under mature trees. Using morphotyping and restriction fragment length polymorphism (RFLP) analysis of the nuclear rDNA internal transcribed spacer (ITS), we were able to distinguish the introduced Thelephoroid fungal sp. ORS.XM002 among native ectomycorrhizal fungal species that form ectomycorrhizae in A. africana seedlings. The morphotype (MT) of the introduced fungus showed some color variation, with a shift from light- to dark-brown observed from younger to older mycorrhizal tips. We were able to differentiate the ITS type xm002 of the introduced fungus from the 14 ITS-RFLP types characterizing the 9 native MT that occurred in forest soils. The frequency of ITS type xm002 ranged from 40% to 49% depending on the forest soil used, and was always higher than those of ITS types from native dark-brown MT that occurred in inoculated seedlings 3 months after inoculation. We considered Thelephoroid fungal sp. ORS.XM002 to be responsible for stimulation of mycorrhizal colonization of inoculated A. africana seedlings when compared with control seedlings in forest soils. This fungus appeared to be more effective in increasing the root dry weight of A. africana seedlings. To identify the unknown introduced fungal species and native MT, we sequenced the ML5/ML6 region of the mitochondrial large subunit rRNA. Sequence analysis showed that these fungi belong to three ML5/ML6 groups closely related to the Cortinarioid, Thelephoroid, and Sclerodermataceous taxa. The molecular evidence for the persistence of Thelephoroid fungal sp. ORS.XM002 despite competition from native fungi argues in favor of using this fungus with A. africana in nursery soil conditions in Senegal.     

Host plants and edaphic factors influence the distribution and diversity of ectomycorrhizal fungal fruiting bodies within rainforests from Tshopo,Democratic Republic of the Congo

Ectomycorrhizal fungi constitute an important component of forest ecosystems that enhances plant nutrition and resistance against stresses. Diversity of ectomycorrhizal (EcM) fungi is, however, affected by host plant diversity and soil heterogeneity. This study provides information about the influence of host plants and soil resources on the diversity of ectomycorrhizal fungal fruiting bodies from rainforests of the Democratic Republic of the Congo. Based on the presence of fungal fruiting bodies, significant differences in the number of ectomycorrhizal fungi species existed between forest stand types (p < 0.001). The most ectomycorrhizal species‐rich forest was the Gilbertiodendron dewevrei‐dominated forest (61 species). Of all 93 species of ectomycorrhizal fungi, 19 demonstrated a significant indicator value for particular forest stand types. Of all analysed edaphic factors, the percentage of silt particles was the most important parameter influencing EcM fungi host plant tree distribution. Both host trees and edaphic factors strongly affected the distribution and diversity of EcM fungi. EcM fungi may have developed differently their ability to successfully colonise root systems in relation to the availability of nutrients.     

Transfer of <Superscript>14</Superscript>C-photosynthate to the sporocarp of an ectomycorrhizal fungus <Emphasis Type="Italic">Laccaria amethystina</Emphasis>

Sporocarps of ectomycorrhizal fungi are strong carbon sinks for the source in host trees, but the details of carbon transfer from the host to the sporocarp are unknown. In this study, single seedlings of Japanese red pine (Pinus densiflora) colonised by Laccaria amethystina were grown on floral foam plates fitted in rhizoboxes, resulting in fruiting on the substrate. The seedlings were photosynthetically labelled with ¹⁴CO₂; ¹⁴C-labelled photosynthate transfer from leaves to sporocarps was then chased using a time-course autoradiography technique. ¹⁴C was transferred to healthy, fresh sporocarps in a purple colour ranging from primordial to elongate sporocarps, but hardly to senesced ones that had faded to white or grey, or browned. This suggested that C is transferred only to physiologically active sporocarps. Two seedlings associated with a growing sporocarp were labelled again 7 and 16 days after the first labelling, respectively. ¹⁴C accumulation in the sporocarps rose in a stepwise manner after the second labelling, indicating that sporocarps mainly used recently rather than previously photosynthesised C.     

Salinity effects on germination,growth, and seed production of the halophyte Cakile maritima

The growth ofEucalyptus regnans seedlings in forest soil is enhanced when it has been air-dried. In undried forest soil seedlings grow poorly and develop purple coloration in the foliage, indicating P deficiency. This paper reports the results of pot experiments designed to investigate the relationship between growth and P acquisition, ectomycorrhizal infection and age of seedlings grown in air-dried and undried soil. The effect on seedling growth of their inoculation with air-dried or undried soil or with ectomycorrhizal roots from plants growing in air-dried or undried soil was also investigated. Ectomycorrhizal root tips were detected in 3-week-oldE. regnans seedlings in both air-dried and undried soil, and from then on the frequency of ectomycorrhizal root tips increased rapidly. In air-dried soil, seedlings were fully ectomycorrhizal at 9 weeks, and the occurrence of maximum ectomycorrhizal infection coincided with enhanced P acquisition and the initiation of rapid seedling growth. In undried forest soil, seedling growth remained poor, even though the seedlings had well-developed ectomycorrhizae and the incidence of ectomycorrhizal root tips was the same as in air-dried soil. The dominant ectomycorrhizae in airdried soil were associated with an ascomycete fungus, whereas in undried, undisturbed soil they were commonly associated with basidiomycete fungi. Inoculation of sterile soil/sand mix with washed ectomycorrhizal roots from air-dried soil increased the P acquisition and growth of the seedlings significantly compared with controls, whereas ectomycorrhizal inocula from undried soil had no effect on seedling growth, although both inocula resulted in a similar incidence of ectomycorrhizal root tips. Similarly, addition of a small amount of air-dried soil into sterile soil/sand mix resulted in a significantly greater increase in the P content and dry weight of the seedlings, compared with the control, than addition of undried soil. In both treatments, the incidence of ectomycorrhial root tips was similar. As (i) the differentiation in seedling growth between air-dried and undried soil occurred after seedlings became ectomycorrhizal, (ii) the dominant ectomycorrhizae in air-dried soil were different from those in undried soil, and (iii) inocula from air-dried soil, but not from undried soil, stimulated seedling growth in sterile soil/sand mix, it is concluded that development of particular ectomycorrhizae may be involved in seedling growth stimulation and enhanced P acquisition associated with air drying of forest soil.     

Spatial structure and diversity of woody plants and ectomycorrhizal fungus sporocarps in a natural subtropical forest

Spatial patterns of ectomycorrhizal fungi, ectomycorrhizal plants, and non-ectomycorrhizal plants were investigated in a natural subtropical forest using second-order analysis. The results of spatial pattern analysis showed that the degree of clumping of woody plants and ectomycorrhizal sporocarps were correlated. There was a significantly positive correlation of relative aggregation indices between ectomycorrhizal fungi and both non-ectomycorrhizal trees and ectomycorrhizal saplings. Correlations between percentage of ectomycorrhizal trees and sporocarp occurrence of ectomycorrhizal fungi and between diversities of woody plants and ectomycorrhizal fungi were distance-dependent or scale-related. A significantly high percentage of ectomycorrhizal trees was found only at relatively short distance from ectomycorrhizal fungal sporocarps, and significantly positive correlation of the diversity between woody plants and ectomycorrhizal fungi was found only at relative long distance, which implied that ectomycorrhizal sporocarps prefer ectomycorrhizal-tree-dominant micro-sites at near distances and at relatively large scales, diverse ectomycorrhizal sporocarps could be found in woodlands with high diversity of woody plants. Important factors affecting the spatial distribution, occurrence, and diversity of ectomycorrhizal fungi include spatial pattern of ectomycorrhizal plants and non-ectomycorrhizal plants, percentage of ectomycorrhizal plants, and plant diversity in a natural forest.     

Multi-host ectomycorrhizal fungi are predominant in a Guinean tropical rainforest and shared between canopy trees and seedlings

The diversity of ectomycorrhizal (ECM) fungi on adult trees and seedlings of five species, Anthonotha fragrans, Anthonotha macrophylla, Cryptosepalum tetraphyllum, Paramacrolobium coeruleum and Uapaca esculenta, was determined in a tropical rain forest of Guinea. Ectomycorrhizae were sampled within a surface area of 1600 m(2), and fungal taxa were identified by sequencing the rDNA Internal Transcribed Spacer region. Thirty-nine ECM fungal taxa were determined, of which 19 multi-hosts, 9 single-hosts and 11 singletons. The multi-host fungi represented 92% (89% when including the singletons in the analysis) of the total abundance. Except for A. fragrans, the adults of the host species displayed significant differentiation for their fungal communities, but their seedlings harboured a similar fungal community. These findings suggest that there was a potential for the formation of common mycorrhizal networks in close vicinity. However, no significant difference was detected for the δ(13)C and δ(15)N values between seedlings and adults of each ECM plant, and no ECM species exhibited signatures of mixotrophy. Our results revealed (i) variation in ECM fungal diversity according to the seedling versus adult development stage of trees and (ii) low host specificity of ECM fungi, and indicated that multi-host fungi are more abundant than single-host fungi in this forest stand.     

Ectomycorrhizal fungal diversity in orchards of cultivated pecan (Carya illinoinensis; Juglandaceae)

Carya illinoinensis (pecan) belongs to the Juglandaceae (walnut family) and is a major economic nut crop in the southern USA. Although evidence suggests that some species in the Juglandaceae are ectomycorrhizal, investigations on their ectomycorrhizal fungal symbionts are quite limited. Here we assessed the ectomycorrhizal fungal diversity in cultivated orchards of C. illinoinensis. Five pecan orchards in southern Georgia, USA, were studied, three of which were known to fruit the native edible truffle species Tuber lyonii. We sequenced rDNA from single ectomycorrhizal root tips sampled from a total of 50 individual trees. Mycorrhizae were identified by ITS and LSU rDNA sequence-based methods. Forty-four distinct ectomycorrhizal taxa were detected. Sequestrate taxa including Tuber and Scleroderma were particularly abundant. The two most abundant sequence types belonged to T. lyonii (17%) and an undescribed Tuber species (~20%). Because of our interest in the ecology of T. lyonii, we also conducted greenhouse studies to determine whether this species would colonize and form ectomycorrhizae on roots of pecan, oak, or pine species endemic to the region. T. lyonii ectomycorrhizae were formed on pecan and oak seedlings, but not pine, when these were inoculated with spores. That oak and pecan seedling roots were receptive to truffle spores indicates that spore slurry inoculation could be a suitable method for commercial use and that, ecologically, T. lyonii may function as a pioneer ectomycorrhizal species for these hosts.     

Slowed decomposition is biotically mediated in an ectomycorrhizal, tropical rain forest

Bacteria and fungi drive the cycling of plant litter in forests, but little is known about their role in tropical rain forest nutrient cycling, despite the high rates of litter decay observed in these ecosystems. However, litter decay rates are not uniform across tropical rain forests. For example, decomposition can differ dramatically over small spatial scales between low-diversity, monodominant rain forests, and species-rich, mixed forests. Because the climatic patterns and soil parent material are identical in co-occurring mixed and monodominant forests, differences in forest floor accumulation, litter production, and decomposition between these forests may be biotically mediated. To test this hypothesis, we conducted field and laboratory studies in a monodominant rain forest in which the ectomycorrhizal tree Dicymbe corymbosa forms >80% of the canopy, and a diverse, mixed forest dominated by arbuscular mycorrhizal trees. After 2 years, decomposition was significantly slower in the monodominant forest (P < 0.001), but litter production was significantly greater in the mixed forest (P < 0.001). In the laboratory, we found microbial community biomass was greater in the mixed forest (P = 0.02), and the composition of fungal communities was distinct between the two rain forest types (P = 0.001). Sequencing of fungal rDNA revealed a significantly lower richness of saprotrophic fungi in the monodominant forest (19 species) relative to the species-rich forest (84 species); moreover, only 4% percent of fungal sequences occurred in both forests. These results show that nutrient cycling patterns in tropical forests can vary dramatically over small spatial scales, and that changes in microbial community structure likely drive the observed differences in decomposition.     

接种彩色豆马勃对模拟酸沉降下马尾松幼苗生物量的影响

外生菌根能够提高宿主植物对外界环境胁迫的抵抗力,促进植物的生长,本文试图揭示外生菌根对酸雨胁迫下马尾松生长的保护作用。本研究采用盆栽试验,共设置四个处理：酸雨对照处理（Control check (CK), 约pH值5.5）不接种,酸雨对照处理接种,酸雨pH值3.5处理不接种,酸雨pH值3.5处理接种。pH值3.5的酸雨处理降低马尾松的生物量,在试验前期降低根冠比,试验中后期则提高根冠比,在试验初期增加了叶面积,但中后期显著降低了叶面积。接种外生菌根菌有利于马尾松幼苗的生长,pH值3.5处理下接种外生菌根菌能提高马尾松幼苗的生物量,外生菌根菌对生物量分配和叶面积的影响与酸雨胁迫的影响是相反的,即外生菌根菌抵消了酸雨胁迫对马尾松的影响。     

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.     

Comparisons of ectomycorrhizae on pinyon pines (Pinus edulis; Pinaceae) across extremes of soil type and herbivory

Using field and greenhouse studies, we examined the relationships among pinyon pines (Pinus edulis), their ectomycorrhizal mutualists, and their moth herbivores as a function of soil fertility. We studied two soil types—the ash and cinder soils of the San Francisco volcanic field and nearby sandy loam soils. In the field, pinyons growing in cinders suffered from reduced moisture, negative nitrogen mineralization rates, low phosphate levels, reduced growth, and high moth herbivory relative to pinyons growing in sandy loam. Pinyons growing in cinders also had twofold higher levels of ectomycorrhizal colonization than their noncinder counterparts. Similarly, in the greenhouse, seedlings grown in cinders had higher levels of ectomycorrhizal colonization and greater numbers of ectomycorrhizae than seedlings grown in sandy loam. Seedling shoot growth was significantly enhanced by ectomycorrhizae in both soils. These patterns support three conclusions. First, field and greenhouse studies demonstrated that trees growing in nutrient-poor soils had higher levels of ectomycorrhizal colonization than trees growing in better soils. Second, across soil types, variation in ectomycorrhizal colonization was better predicted by soil fertility than by herbivory. However, herbivory negatively affected ectomycorrhizae in the stressful cinder environment. Third, although mycorrhizae can be parasitic under some conditions, ectomycorrhizae had mutualistic impacts on pinyon seedlings across the environmental extremes we studied.     

Diversity and community composition of ectomycorrhizal fungi in a dry deciduous dipterocarp forest in Thailand

Large forest areas of South-East Asia, are dominated by the Dipterocarpaceae tree family, which contains many important timber species. Unlike many other tropical trees, Dipterocarpaceae rely on ectomycorrhizal (ECM) root symbiosis for their mineral nutrition. This study aims to document the richness and community composition of ECM fungi in a dry deciduous forest in Thailand. Combining morphological and molecular identification methods revealed 69 species of ECM fungi that belong to 17 phylogenetic lineages. The /russula-lactarius, /tomentella-thelephora, /sordariales, /sebacina and /cantharellus lineages were the most species-rich. The fungal richness is comparable to other tropical rain forest sites, but the phylogenetic community structure has elements of both tropical and temperate ecosystems. Unlike tropical rain forests, the Cenococcum geophilum complex was one of the most frequent fungal taxa that had a relatively high ITS genetic diversity over the small sampling area. This study provides the first snapshot insight into the fungal community of dry dipterocarp forests. However, it is necessary to broaden the spatial and temporal scales of sampling to improve our understanding of the below-ground relations of dry and humid tropical forests.     

外生菌根缓解植物酸雨胁迫的机理研究进展

森林作为陆地生态系统的主体,是酸雨污染的主要受体,酸雨对生态系统产生着巨大的影响。菌根是菌根真菌与植物营养根的共生体。外生菌根真菌与宿主植物间互惠互利,在森林生态系统中,外生菌根在维持生态系统的养分平衡和改善树木营养等方面有重要的作用。本文综述了国内外关于菌根和酸雨关系的研究,酸雨能抑制外生菌根的形成,降低其活力;但另一方面,外生菌根能够缓解酸雨造成的植物危害,提高植株对酸雨的耐受力。外生菌根主要通过以下几方面缓解酸雨胁迫:(1)菌根形态结构的物理屏蔽作用;(2)增加养分吸收,增加御酸能力;(3)增强酶活性,提高植物生存能力;(4)产生有机酸或其他物质。     

EcM fungal community structure, but not diversity, altered in a Pb-contaminated shooting range in a boreal coniferous forest site in Southern Finland

Boreal forests contain diverse fungal communities that form essential ectomycorrhizal symbioses with trees. To determine the effects of lead (Pb) contamination on ectomycorrhizal fungal communities associated with the dominant pine (Pinus sylvestris L.), we surveyed sporocarps for 3 years, analyzed morphotyped ectomycorrhizal root tips by direct sequencing, and 454-sequenced fungal communities that grew into in-growth bags during a 2-year incubation at a shooting range where sectors vary in the Pb load. We recorded a total of 32 ectomycorrhizal fungi that formed conspicuous sporocarps, 27 ectomycorrhizal fungal phylotypes from 294 root tips, and 116 ectomycorrhizal fungal operation taxonomic unit (OTUs) from a total of 8194 internal transcribed spacer-2 454 sequences. Our ordination analyses by nonparametric multidimensional scaling (NMS) indicated that the Pb enrichment induced a shift in the ectomycorrhizal community composition. This was visible as indicative trends in the sporocarp and root tip data sets, but was explicitly clear in the communities observed in the in-growth bags. The compositional shift in the ectomycorrhizal community was mainly attributable to an increase in the frequencies of OTUs assigned to genus Thelephora and to a decrease in the OTUs assigned to Pseudotomentella, Suillus, and Tylospora in Pb-contaminated areas when compared with the control. While the compositional shifts are clear, their functional consequences for the dominant trees or soil ecosystem function remain undetermined.     

Fungal content of ectomycorrhizal tips: comparison among 13 tree species

To better understand soil carbon cycling in forest ecosystems, we studied the proportion of fungal sheath area (FSA) in the cross-sectional ectomycorrhizal area in 13 tree species. Ectomycorrhizal samples were collected from subalpine and temperate forests in Japan. The FSA values were in the range of 12% to 56% across all tree species, tree ages, and fungal species. In Abies firma and Quercus serrata, the FSA values were larger in mature trees than in seedlings, whereas no such differences were found in Pinus densiflora and Fagus crenata. In broad-leaved trees, because the plant tissue radii lay within a narrow range, the FSA was affected mainly by the fungal sheath thickness. In conifers, however, the plant tissue radii varied widely among genera, so the FSA was affected by both the plant tissue radius and the fungal sheath thickness. Our findings suggest that the fungal content of ectomycorrhizal tips differs among tree species and fungal species, so that both parameters must be considered in studies of forest carbon cycling. The estimates revealed that data gathering in each type of forest leads to more accurate estimates of the biomass of fungi in ectomycorrhizal tips.