Interactions between extraradical ectomycorrhizal mycelia, microbes associated with the mycelia and growth rate of Norway spruce (Picea abies) clones

Despite their ecological relevance, field studies of the extraradical mycelia of ectomycorrhizal (ECM) fungi are rare. Here we examined in situ interactions between ECM mycelia and host vigour. Ectomycorrhizal mycelia were harvested with in-growth mesh bags buried under Norway spruce (Picea abies) clones planted in 1994 in a randomized block design. Mycelial biomass was determined and fungal species were identified by denaturing gradient gel electrophoresis (DGGE) and sequencing of the internal transcribed spacer 1 (ITS1) region. Microbial community structure in the mycelium was investigated by phospholipid fatty acid (PLFA) profiling. Compared to slow-growing spruce clones, fast-growing clones tended to support denser mycelia where the relative proportions of Atheliaceae fungi and PLFAs indicative of Gram-positive bacteria were higher. Ascomycetes and PLFAs representative of Gram-negative bacteria were more common with slow-growing clones. In general, the ECM mycelial community was similar to the ECM root-tip community. Growth rate of the hosts, the ECM mycelial community and the microbes associated with the mycelium were related, suggesting multitrophic interactions between trees, fungi and bacteria.     

Ectomycorrhizal fungi: the symbiotic route to the root for phosphorus in forest soils

Many forest trees have evolved mutualistic symbioses with ectomycorrhizal (ECM) fungi that contribute to their phosphorus (P) nutrition. Forest productivity is frequently limited by P, a phenomenon that is likely to become more widespread under future conditions of elevated atmospheric CO₂ concentration [CO₂]. It is thus timely that this review considers current understanding of the key processes (absorption, translocation and transfer to the plant host) in ECM fungus-mediated P nutrition of forest trees. Solubilisation of inorganic P (P_i) and hydrolysis of organic P by ECM fungi in soil occurs largely at the growing mycelial front, where P_i absorption is facilitated by high affinity transporters. While large gaps remain in our understanding of the physiological and molecular mechanisms that underpin movement of P in ECM mycelia in soil and P transfer to the plant, host P demand seems likely to be a key driver of these processes. ECM fungi may make considerable contributions to meeting the likely increased P demand of trees under elevated [CO₂] via increased colonization levels, shifts in ECM fungal community structure and changed patterns of EMM production. Further research into the spatial scale of ECM-mediated P movements in soil, along with the interplay between ECM fungi and other soil microflora is advocated.     

Influence of long-term repeated prescribed burning on mycelial communities of ectomycorrhizal fungi

To demonstrate the efficacy of direct DNA extraction from hyphal ingrowth bags for community profiling of ectomycorrhizal (ECM) mycelia in soil, we applied the method to investigate the influence of long-term repeated prescribed burning on an ECM fungal community. DNA was extracted from hyphal ingrowth bags buried in forest plots that received different prescribed burning treatments for 30 yr, and denaturing gradient gel electrophoresis (DGGE) profiles of partial fungal rDNA internal transcribed spacer (ITS) regions were compared. Restriction fragment length polymorphism (RFLP) and sequence analyses were also used to compare clone assemblages between the treatments. The majority of sequences derived from the ingrowth bags were apparently those of ECM fungi. DGGE profiles for biennially burned plots were significantly different from those of quadrennially burned and unburned control plots. Analysis of clone assemblages indicated that this reflected altered ECM fungal community composition. The results indicate that hyphal ingrowth bags represent a useful method for investigation of ECM mycelial communities, and that frequent long-term prescribed burning can influence below-ground ECM fungal communities.     

Is plant genetic control of ectomycorrhizal fungal communities an untapped source of stable soil carbon in managed forests?

Background

Ectomycorrhizal (ECM) fungi provide one of the main pathways for carbon (C) to move from trees into soils, where these fungi make significant contributions to microbial biomass and soil respiration.

Scope

ECM fungal species vary significantly in traits that likely influence C sequestration, such that forest C sequestration potential may be driven in part by the existing community composition of ECM fungi. Moreover, accumulating experimental data show that tree genotypes differ in their compatibility with particular ECM fungal species, i.e. mycorrhizal traits of forest trees are heritable. Those traits are genetically correlated with other traits for which tree breeders commonly select, suggesting that selection for traits of interest, such as disease resistance or growth rate, could lead to indirect selection for or against particular mycorrhizal traits of trees in forest plantations.

Conclusions

Altogether, these observations suggest that selection of particular tree genotypes could alter the community composition of symbiotic ECM fungi in managed forests, with cascading effects on soil functioning and soil C sequestration.     

四川都江堰亚热带针阔混交林中主要晚生型外生菌根真菌类群的空间分布

晚生型外生菌根真菌通常出现在森林演替的后期,是成熟林中的优势外生菌根真菌类群.对四川都江堰一片亚热带针阔混交林中的菌根真菌地上群落进行调查,并应用二元逻辑回归分析对晚生型外生菌根真菌的主要类群,即鹅膏菌科、牛肝菌科和红菇科,与周围(5 m×5 m样方)树种组成的关系进行研究.还应用次级变量分析方法对主要外生菌根真菌类群的空间格局进行了分析.结果表明,非外生菌根树种及某些外生菌根树种对特定类群菌根真菌子实体的出现有抑制作用,而不同类群外生菌根真菌在克隆生长上的差异并不是子实体空间分布的决定因素.我们认为,当研究自然林中外生菌根子实体的空间分布时,除了宿主植物的分布,也应考虑非宿主植物的分布以及菌根真菌相互作用的影响.     

白皮松天然林外生菌根土壤繁殖体库

为更好地恢复和保存白皮松天然林,在陕西省白皮松残存林地采集根际土壤,采用幼苗检测法获取土壤外生菌根真菌繁殖体,用形态观察分类与ITS-PCR-sequencing相结合的方法进行菌根鉴定,研究白皮松林地外生菌根真菌土壤繁殖体库的组成.结果表明: 在白皮松幼苗菌根中共获得73个特异性序列;按照97%的相似度阈值,将序列划分为12个可操作分类单元(OTUs);稀疏曲线分析表明,本研究基本获得了白皮松土壤外生菌根繁殖体库的多样性.常见种有土生空团菌、Tomentella sp.、Tuber sp.等.出现频率最高的一个OTU(80%)与已知种类相似度较低(75%),说明其可能是一个新的菌根菌种类.白皮松残存天然林地的外生菌根繁殖体库中具有其他松科植物土壤繁殖体库常见的种类,但是频率最高的种类未能鉴定到已知属或科,说明白皮松菌根繁殖体库具有其宿主特异性.这种群落特异构成也说明研究和利用白皮松土壤外生菌根真菌繁殖体库具有特殊性和重要性.     

Tree‐mycorrhizal associations detected remotely from canopy spectral properties

A central challenge in global ecology is the identification of key functional processes in ecosystems that scale, but do not require, data for individual species across landscapes. Given that nearly all tree species form symbiotic relationships with one of two types of mycorrhizal fungi – arbuscular mycorrhizal (AM) and ectomycorrhizal (ECM) fungi – and that AM‐ and ECM‐dominated forests often have distinct nutrient economies, the detection and mapping of mycorrhizae over large areas could provide valuable insights about fundamental ecosystem processes such as nutrient cycling, species interactions, and overall forest productivity. We explored remotely sensed tree canopy spectral properties to detect underlying mycorrhizal association across a gradient of AM‐ and ECM‐dominated forest plots. Statistical mining of reflectance and reflectance derivatives across moderate/high‐resolution Landsat data revealed distinctly unique phenological signals that differentiated AM and ECM associations. This approach was trained and validated against measurements of tree species and mycorrhizal association across ~130 000 trees throughout the temperate United States. We were able to predict 77% of the variation in mycorrhizal association distribution within the forest plots (P < 0.001). The implications for this work move us toward mapping mycorrhizal association globally and advancing our understanding of biogeochemical cycling and other ecosystem processes.     

寒温带地区典型森林类型中外生菌根真菌多样性和群落组成及其影响因素

外生菌根(ECM)真菌可促进宿主植物对土壤中水分和营养的吸收,提高宿主植物的抗逆性及对环境的适应,在森林生态系统的物质循环中发挥了重要作用。为探明寒温带地区ECM真菌多样性和群落组成以及土壤理化因子对真菌群落组成的可能影响机制,采用高通量测序(Illumina Miseq)平台对大兴安岭4种典型森林类型白桦、樟子松、兴安落叶松和偃松的根围土壤ECM真菌群落组成进行了分析。结果表明,4种典型森林类型根围土壤中共获得167个ECM真菌可操作分类单元(OTUs),隶属于2个门、4个纲、10个目、24个科、36个属。在门水平上,担子菌门(Basidiomycota)真菌的相对丰度占96.4%,子囊菌门(Ascomycota)真菌占3.6%。在属水平上,红菇属(Russula,30.2%)、丝膜菌属(Cortinarius,17.1%)、Piloderma(9.6%)、棉革菌属(Tomentella,7.2%)、丝盖伞属(Inocybe,7.2%)和蜡壳耳属(Sebacina,6.2%)的相对丰度均>5.0%,是该地区的优势ECM真菌类群。不同森林类型的优势ECM真菌在属水平上的组成存在显...     

四川都江堰亚热带针阔混交林中主要晚生型外生菌根真菌类群的空间分布

晚生型外生菌根真菌通常出现在森林演替的后期,是成熟林中的优势外生菌根真菌类群。对四川都江堰一片亚热带针阔混交林中的菌根真菌地上群落进行调查,并应用二元逻辑回归分析对晚生型外生菌根真菌的主要类群,即鹅膏菌科、牛肝菌科和红菇科,与周围(5m×5m样方)树种组成的关系进行研究。还应用次级变量分析方法对主要外生菌根真菌类群的空间格局进行了分析。结果表明,非外生菌根树种及某些外生菌根树种对特定类群菌根真菌子实体的出现有抑制作用,而不同类群外生菌根真菌在克隆生长上的差异并不是子实体空间分布的决定因素。我们认为,当研究自然林中外生菌根子实体的空间分布时,除了宿主植物的分布,也应考虑非宿主植物的分布以及菌根真菌相互作用的影响。     

The role of fungi in biogenic weathering in boreal forest soils

In this article we discuss the possible significance of biological processes, and of fungi in particular, in weathering of minerals. We consider biological activity to be a significant driver of mineral weathering in forest ecosystems. In these environments fungi play key roles in organic matter decomposition, uptake, transfer and cycling of organic and inorganic nutrients, biogenic mineral formation, as well as transformation and accumulation of metals. The ability of lichens, mutualistic symbioses between fungi and photobionts such as algae or cyanobacteria, to weather minerals is well documented. The role of mycorrhizal fungi forming symbioses with forest trees is less well understood, but the mineral horizons of boreal forests are intensively colonised by mycorrhizal mycelia which transfer protons and organic metabolites derived from plant photosynthates to mineral surfaces, resulting in mineral dissolution and mobilisation and redistribution of anionic nutrients and metal cations. The mycorrhizal mycelia, in turn provide efficient systems for the uptake and direct transport of mobilised essential nutrients to their host plants which are large sinks. Since almost all (99.99 %) non-suberised lateral plant roots involved in nutrient uptake are covered by ectomycorrhizal fungi, most of this exchange of metabolites must take place through the plant–fungus interface. This idea is still consistent with a linear relationship between soil mineral surface area and weathering rate since the mycelia that emanate from the tree roots will have a larger area of contact with minerals if the mineral surface area is higher. Although empirical models based on bulk soil solution chemistry may fit field data, we argue that biological processes make an important contribution to mineral weathering and that a more detailed mechanistic understanding of these must be developed in order to predict responses to environmental changes and anthropogenic impact.     

Localisation of phosphomonoesterase activity in ectomycorrhizal fungi grown on different phosphorus sources

Phosphorus (P) is a major limiting nutrient for plants in boreal forest ecosystems where a substantial part of the total P is sequestered in organic compounds. Some ectomycorrhizal (ECM) fungi are known to produce phosphomonoesterases, enzymes that degrade organic P sources. Here, we test 16 ECM species for this enzymatic activity by growing them on media containing orthophosphate, phytic acid or apatite. A method with an overlay gel that determined both phosphomonoesterase activity and its spatial distribution was developed. The phosphomonoesterase activity was not significantly higher when growing on organic P; conversely some isolates only produced measurable enzyme activity when grown on apatite. Species-specific variations with respect to phosphomonoesterase activity as well as growth responses to different substrates were found. The production of phosphomonoesterases was found to be widespread in ECM fungi and the enzyme activity did not need induction by organic P. The enzyme activity was highest in the central parts of the mycelia, potentially reflecting breakdown and recycling of phospholipids from old hyphae or potentially higher mycelial density.     

Spatial distribution of ectomycorrhizal mats in coniferous forests of the Pacific Northwest, USA

Ectomycorrhizal mats in forest soils have a wide global distribution and have been noted as potentially important elements in forest soil nutrient cycling. To elucidate the relationship between ectomycorrhizal mats and their environment, we undertook field studies and spatial analyses of mat distributions at different spatial scales.We used two experimental approaches to study mat-forming ectomycorrhizal fungi in coniferous forests of the Pacific Northwest in the United States. In the first approach, ectomycorrhizal mats and other forest floor features were mapped in 2 × 10 m plots and digitized into a geographical information system (GIS) for spatial pattern analysis. In order to examine larger-scale phenomena, a second approach involving other sites was taken; soil cores were taken along 30-m transects, and distance to the closest living potential host tree was calculated for each core.Mat patterns were studied at two scales: (1) within-stand level (i.e. variability attributed to distribution of other mat species, forest floor attributes, and understory vegetation); and (2) stand level (i.e. variability expressed along a successional gradient). Mat distribution was influenced by: (1) the proximity of one mat to another; (2) the distance from the mat to the closest living tree; (3) the density of living trees in a stand; and (4) the successional stage of the stand.Although GIS analysis indicated that mats of different morphologies did not physically overlap, there was a tendency for clustering of mats. No apparent correlations were observed between forest floor features and mats located within the 2 × 10 m grids. On the scale of tens of meters, mats decreased with distance from the closest potential host tree. Spatial patterns of mat distributions in harvested sites suggest that these mats may persist at least 2 years after their host trees have been cut. For Gautieria mats, total mat area, size, and frequency differed with stand age.This study has demonstrated the importance of both spatial scaling and forest stand age when the natural distribution of mycorrhizal fungi is examined. Results suggest the need for mat research directed at higher-order scales (e.g. stand and watershed) that will provide accurate information for managing forests to ensure their survival and normal function. ei]J H Graham     

Ectomycorrhizal fungi and their enzymes in soils: is there enough evidence for their role as facultative soil saprotrophs?

Although ectomycorrhizal (ECM) fungi are generally regarded as dependent upon the supply of carbon from their plant hosts, some recent papers have postulated a role for these fungi in the saprotrophic acquisition of carbon from soil. This theory was mainly based on the increase in enzymatic activity during periods of low photosynthate supply from tree hosts and emergence of the theory has led to a question about the overall influence of saprotrophy by ECM fungi on soil carbon turnover. However, I argue here that there is still not enough evidence to confirm this proposed function. My argument is based on inference from several lines of observation and concern over several aspects of the past studies. First, ECM fungi mainly inhabit deeper soil horizons, in which the availability of carbon compounds with positive energetic value is low. Second, the ability of ECM fungi to produce ligninolytic enzymes and cellulases is much weaker than that of saprotrophic basidiomycetes. This is most apparent in the low copy abundance of corresponding genes in the sequenced genomes of ECM species Laccaria bicolor and Amanita bisporigenes compared to the saprotrophic species Galerina marginata. I offer alternative hypotheses to explain the past observations of increased enzyme activity during starvation periods. These include, the induction of autolytic processes in ECM fungal mycelia or an attack on the host tissues to support escape from a dying root and to allow for a search for new hosts.     

Structure and species composition of ectomycorrhizal fungal communities colonizing seedlings and adult trees of Pinus montezumae in Mexican neotropical forests

Mexico is a center of diversity for pines, but few studies have examined the ectomycorrhizal (ECM) fungal communities associated with pines in this country. We investigated the ECM communities associated with Pinus montezumae seedlings and mature trees in neotropical forests of central Mexico and compared their structure and species composition. Root tips were sampled on both planted seedlings and naturally occurring adult trees. A total of 42 ECM operational taxonomic units (OTUs) was found on P.?montezumae. Diversity and similarity indices showed that community structure was similar for both plant growth stages, but phylogenetic diversity and Chao-estimated richness were higher for seedlings. Species composition differed between communities. The dominant OTUs belonged to the families Atheliaceae, Cortinariaceae, and Sebacinaceae, although different taxa appeared to colonize seedlings and adults. Only 12 OTUs were shared between seedlings and adults, which suggests that ECM fungi which colonize seedlings are still not fully incorporated into mycelial networks and that ECM taxa colonizing young individuals of P.?montezumae are likely to come from fungal propagules. Intra-generic diversity could be an insurance mechanism to maintain forest productivity under stressed conditions. This is the first report describing the abundance of Atheliaceae in tree roots in neotropical ecosystems.     

Identification of the DNAs of the ectomycorrhizal fungusTricholoma bakamatsutake using specific oligonucleotide probes and PCR primers

Partial nucleotides of the 18S rDNAs ofTricholoma bakamatsutake were sequenced and compared with those of six ectomycorrhizal fungi and a tree. Two probes, Probes 1 and 2, and a pair of primers were designed based on the variable positions in this region. The DNAs ofT. bakamatsutake were isolated from the colonized mycelia in the soil, field-collected fruit-bodies and artifically cultured mycelia. Hybridization with Probe 1 and PCR-amplification with the primers differentiated these DNAs of this fungus from those of eight ectomycorrhizal fungi and two tree species.     

Pioneer dwarf willow may facilitate tree succession by providing late colonizers with compatible ectomycorrhizal fungi in a primary successional volcanic desert

To advance understanding of the contribution of ectomycorrhizal (ECM) fungi to tree successional processes, natural establishment patterns of secondary colonizing hosts and their ECM fungal communities were investigated with special reference to pioneer hosts. In the volcanic desert on Mount Fuji, Japan, vegetation is sparsely distributed, resembling islands in a sea of scoria. Of 509 vegetation islands in the research area, 161 contained Salix reinii (Salix), the first colonizing ECM host species. The spatial coincidence between secondary colonizing timber species and Salix was analysed, and ECM fungal communities were studied using molecular identification methods. I found 39 and 26 individuals of Betula ermanii and Larix kaempferi, respectively. Without exception, these individuals were all accompanied by Salix. The ECM fungal communities of these timber species showed high similarity to that of Salix and were dominated by generalists that were compatible with two or more plant families. In this desert, available ECM propagules are limited. Pioneer Salix may contribute to tree succession by providing adjacent late colonizers with compatible ECM fungal symbionts.     

粘盖乳牛肝菌在低磷、酸铝胁迫下的生长和代谢响应

低磷和酸铝胁迫是酸性土壤限制植物生长的主要因素。有研究指出外生菌根(ectomycorrhiza,ECM)可提高宿主植物对铝毒害和低磷胁迫的适应性。然而,目前有关ECM真菌自身对低磷和酸铝环境的适应机理还不清楚。基于此,本研究以我国南方酸性土壤广泛分布的ECM真菌——粘盖乳牛肝菌Suillus bovinus为研究对象,在纯培养条件下研究了低磷、酸铝胁迫对其生长、营养吸收及菌丝分泌物的影响。结果表明,粘盖乳牛肝菌是一种耐铝型真菌,酸铝胁迫(1 mmol/L)不影响其菌丝生长,而低磷胁迫(20 μmol/L)则显著限制其菌丝生长(P<0.05)。值得注意的是,低磷胁迫的抑制效应可被酸铝胁迫逆转。低磷胁迫显著降低了粘盖乳牛肝菌对磷的吸收(P<0.05),而酸铝胁迫则对菌丝钾的吸收有促进作用。低磷、酸铝胁迫同样改变了菌丝分泌物组成。在低磷胁迫下,大量酚酸类、有机酸及脂质代谢物的积累量下调;而酸铝胁迫下则有大量酚酸类物质上调,有机酸和脂质中上调代谢物数量也高于下调数量;低磷酸铝复合胁迫下酚酸和有机酸类代谢物积累量均显著上调。另外,吲哚-3-乙酸(IAA)在各胁迫下均显著上调。以上结果可在一定程度上解释粘盖乳牛菌对低磷、酸铝环境的适应机理,并对后续进一步阐明ECM的共生适应机理有一定指导意义。     

野外模拟酸雨胁迫下接种外生菌根真菌对马尾松幼苗的缓解作用

外生菌根真菌能够提高宿主植物对外界环境胁迫的抵抗力。主要探讨野外条件下外生菌根真菌对酸雨胁迫下马尾松(Pinus massoniana)幼苗生长、养分元素以及表层土壤的影响,以期为酸雨严重区马尾松林恢复提供科学依据。以2年生马尾松幼苗为材料,采用原位试验,共设置6个处理:p H5.6(对照)处理未接种、对照处理接种、p H4.5酸雨处理未接种、p H4.5酸雨处理接种、p H3.5酸雨处理未接种、p H3.5酸雨处理接种。研究表明:(1)酸雨处理与对照处理相比显著降低了非菌根苗总生物量及各部位生物量(根、茎、叶),对株高无显著影响,接种外生菌根真菌可以缓解酸雨对马尾松幼苗生长的不利影响;(2)与对照处理相比,酸雨处理的非菌根苗的针叶中N、P、Ca含量升高,Mg含量降低,根系中N、P、Ca含量降低,Mg含量随p H的降低先升高后降低。接种外生菌根真菌显著提高了p H3.5酸雨处理的马尾松幼苗根系中N、P、Ca、Mg含量,而对针叶中N、P、Ca、Mg含量无显著影响。(3)在非菌根土壤中,p H3.5酸雨处理与对照处理相比显著降低了土壤中有机质、速效磷、速效钾、可溶性碳、可溶性氮、铵态氮、硝态氮含量,而接种外生菌根真菌显著提高了上述指标。酸雨对土壤阳离子交换量无显著影响。总而言之,接种外生菌根真菌促进了酸雨处理的马尾松幼苗生长、缓解了酸雨对马尾松幼苗养分元素和表层土壤的不利影响,由此可见接种外生菌根真菌是减轻酸雨对马尾松危害的一个重要途径。     

Wood ash application in a managed Norway spruce plantation did not affect ectomycorrhizal diversity or N retention capacity

Ectomycorrhizal (ECM) fungi are key players in N cycling in coniferous forests, and forest management such as application of wood ash can affect their functionality. The aim of this study was to determine the effects of wood ash application on ECM fungal mycelial production, capacity to retain N, diversity and community composition. In-growth mesh bags were installed in control and treated plots. After 6 months, ¹⁵N labeled ammonium and nitrate were applied into the mesh bags, and 24 h later extramatrical mycelium (EMM) was extracted and analyzed. Wood ash had no effects on EMM in-growth, N retention capacity, diversity or community composition. In contrast, there were significant seasonal differences in the amount of EMM produced. These results demonstrate that applying up to 6 t ha⁻¹ of wood ash in this type of plantation forest is a safe management practice that does not increase N leaching or negatively affect ECM fungi.