Pine microsatellite markers allow roots and ectomycorrhizas to be linked to individual trees

Linking roots and ectomycorrhizas (EcM) to individual host trees in the field is required to test whether individual trees support different ectomycorrhizal communities. Here we describe a method that identifies the source of EcM roots by PCR of polymorphic pine nuclear microsatellite loci using fluorescently labelled primers and high-throughput fragment analysis. ITS-PCR can also be performed on the same EcM DNA extract for fungal identification. The method was tested on five neighbouring Scots pine (Pinus sylvestris var scotica) trees in native woodland. Successful host tree identification from DNA extracts of EcM root tips was achieved for 93% of all root fragments recovered from soil cores. It was estimated that each individual mature pine sampled was colonised by between 15 and 19 EcM fungi. The most abundant fungal species were found on all five trees, and within the constraints of the sampling scheme, no differences between trees in EcM fungal community structure or composition were detected.     

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.     

Fine-scale distribution of pine ectomycorrhizas and their extramatrical mycelium

In order to clarify the functional role of individual ectomycorrhizal (EcM) fungal species in the field, we need to relate their abundance and distribution as mycorrhizas to their abundance and distribution as extramatrical mycelium (EMM). We divided each of four 20 cm x 20 cm x 2 cm slices of pine forest soil into 100 cubes of 2 cm x 2 cm. For each cube, ectomycorrhizas were identified and the presence of EMM of the EcM fungi recorded as ectomycorrhizas was determined by terminal restriction fragment length polymorphism (T-RFLP) analysis of ITS rDNA. Ectomycorrhizas and EMM of seven EcM species were mapped. Spatial segregation of mycorrhizas and EMM was evident and some species produced their EMM in different soil layers from their mycorrhizas. The spatial relationship between mycorrhizas and their EMM generally conformed to their reported exploration types, but EMM of smooth types (e.g. Lactarius rufus) was more frequent than expected. Different EcM fungi foraged at different spatial scales.     

Towards global patterns in the diversity and community structure of ectomycorrhizal fungi

Global species richness patterns of soil micro-organisms remain poorly understood compared to macro-organisms. We use a global analysis to disentangle the global determinants of diversity and community composition for ectomycorrhizal (EcM) fungi-microbial symbionts that play key roles in plant nutrition in most temperate and many tropical forest ecosystems. Host plant family has the strongest effect on the phylogenetic community composition of fungi, whereas temperature and precipitation mostly affect EcM fungal richness that peaks in the temperate and boreal forest biomes, contrasting with latitudinal patterns of macro-organisms. Tropical ecosystems experience rapid turnover of organic material and have weak soil stratification, suggesting that poor habitat conditions may contribute to the relatively low richness of EcM fungi, and perhaps other soil biota, in most tropical ecosystems. For EcM fungi, greater evolutionary age and larger total area of EcM host vegetation may also contribute to the higher diversity in temperate ecosystems. Our results provide useful biogeographic and ecological hypotheses for explaining the distribution of fungi that remain to be tested by involving next-generation sequencing techniques and relevant soil metadata.     

Ectomycorrhiza succession patterns in Pinus sylvestris forests after stand-replacing fire in the Central Alps

Fires shape fundamental properties of many forest ecosystems and climate change will increase their relevance in regions where fires occur infrequently today. In ecosystems that are not adapted to fire, post-fire tree recruitment is often sparse, a fact that might be attributed to a transient lack of mycorrhizae. Ectomycorrhizal (EcM) fungi play an important role for recruitment by enhancing nutrient and water uptake of their hosts. The questions arise whether and for how long the EcM community is transformed by fire. We investigated the resistance and resilience of EcM fungal communities on a chronosequence of 12 Pinus sylvestris stands in Valais (Switzerland) and Val d’Aosta (Italy) affected by fire between 1990 and 2006. Soil samples from burnt and non-burnt forests were analyzed with respect to EcM fungi by means of a bioassay. The number of EcM species was significantly lower in samples from recently (2–5 years) burnt sites than non-burnt forest, and increased with time since fire reaching levels of adjacent forests after 15–18 years. Community composition changed after fire but did not converge to that of non-burnt sites over the 18 year period. Only Rhizopogon roseolus and Cenococcum geophilum were abundant in both burnt sites and adjacent forest. Our data indicate fire resistance of some EcM fungal species as well as rapid resilience in terms of species number, but not in species composition. As long as the function of different EcM species for seedling establishment is unknown, the consequences of long-term shifts in EcM community composition for tree recruitment remain unclear.     

菌根真菌影响森林生态系统碳循环研究进展

森林生态系统在全球碳(C)储量中占据极为重要的地位。菌根真菌广泛存在于森林生态系统中,在森林生态系统C循环过程中发挥重要的作用。阐述了不同菌根类型真菌在森林生态系统C循环过程中的功能,对比了温带/北方森林与热带/亚热带森林中菌根真菌介导的C循环研究方面新近取得的研究结果。发现温带和北方森林的外生菌根(EcM)植物对地上生物量C的贡献相对较小,然而是地下C储量的主要贡献者;以丛枝菌根(AM)共生为主的热带/亚热带森林地表生物量占比较高,表明AM植被对热带/亚热带森林地上生物量C的贡献相对较大。我们还就全球变化背景下,菌根真菌及其介导的森林生态系统C汇功能,以及不同菌根类型树种影响C循环的机制等进行了总结。菌根真菌通过影响凋落物分解、土壤有机质形成及地下根系生物量,进而影响整个森林生态系统的C循环功能。菌根介导的森林C循环过程很大程度上取决于(优势)树木的菌根类型和森林土壤中菌根真菌的群落结构。最后指出了当前研究存在的主要问题以及未来研究展望。本文旨在明确菌根真菌在森林生态系统C循环转化过程中的重要生态功能,有助于准确地评估森林生态系统C汇现状,为应对全球变化等提供重要的依据。     

Seedling emergence and establishment of Pinus sylvestris in the Mongolian forest-steppe ecotone

The potential of Scots pine (Pinus sylvestris) for regeneration and encroachment onto dry grasslands in the forest-steppe ecotone was experimentally studied at the south-eastern distribution limit of the species in Mongolia. The experiment consisted of a sowing and planting (2-year old seedlings) assay at two different distances from the forest line and manipulation of the water supply by irrigation in one half of the replicate plots. Seedling emergence was strongly inhibited by post-dispersal seed predation (between 50 and >90 % of the seeds were apparently consumed) and by drought. Seedling survival was limited by drought and phytopathogenic fungi that infected the needles. Herbivory by insects or rodents, a key factor for seedling mortality in Larix sibirica, the most frequent tree species in Mongolia’s forest-steppe ecotone, was of little importance in Scots pine, probably due to the production of efficient allelochemicals. The potential of Scots pine to regenerate in Mongolia’s forest-steppe ecotone and to encroach onto the steppe is very limited and mostly restricted to the immediate vicinity of the forest; it might even decrease in future in the face of climate warming. The observed dependence of seedling emergence and survival on soil moisture suggests that regeneration outside the forest may only be successful in exceptionally moist years. Livestock grazing is certainly an additional limiting factor for Scots pine regeneration in Mongolia, but was not relevant in the present study area.     

Ectomycorrhizal fungi of exotic pine plantations in relation to native host trees in Iran: evidence of host range expansion by local symbionts to distantly related host taxa

Introduction of exotic plants change soil microbial communities which may have detrimental ecological consequences for ecosystems. In this study, we examined the community structure and species richness of ectomycorrhizal (EcM) fungi associated with exotic pine plantations in relation to adjacent native ectomycorrhizal trees in Iran to elucidate the symbiont exchange between distantly related hosts, i.e. Fagales (Fagaceae and Betulaceae) and Pinaceae. The combination of morphological and molecular identification approaches revealed that 84.6 % of species with more than one occurrence (at least once on pines) were shared with native trees and only 5.9 % were found exclusively on pine root tips. The community diversity of ectomycorrhizal fungi in the pine plantations adjacent to native EcM trees was comparable to their adjacent native trees, but the isolated plantations hosted relatively a species-poor community. Specific mycobionts of conifers were dominant in the isolated plantation while rarely found in the plantations adjacent to native EcM trees. These data demonstrate the importance of habitat isolation and dispersal limitation of EcM fungi in their potential of host range expansion. The great number of shared and possibly compatible symbiotic species between exotic Pinaceae and local Fagales (Fagaceae and Betulaceae) may reflect their evolutionary adaptations and/or ancestral compatibility with one another.     

The threatened plant intermediate wintergreen (<Emphasis Type="Italic">Pyrola media</Emphasis>) associates with a wide range of biotrophic fungi in native Scottish pine woods

The plant intermediate wintergreen (Pyrola media, Ericaceae) is in need of conservation action in Scotland. Although widespread, it is locally distributed in dwarf shrub heath and more commonly in Scots pine (Pinus sylvestris) woodlands. A recent study on the mycorrhizal status of Pyrola suggested that they associate with a restricted range of ectomycorrhizal (ECM) fungi. Here, we examined the hypothesis that specialisation by P. media for fungi usually associated with Scots pine is a factor in promoting its occurrence in this habitat. The fungal community associated with the roots of P. media growing in a Scots pine forest was determined by morphotyping, polymerase chain reaction, cloning and sequencing. Molecular identification found 49 taxa representing ecto- and ericoid mycorrhizal fungi, dark septate endophytes, saprotrophs, and fungi of unknown trophic status. The majority of the taxa (67.4%) were Basidiomycota, with 24.4% known to be ECM fungi specific to Pinus sp. or conifers. However, a wide range of other mycorrhizal fungi with varying degrees of host specificity were also found, including taxa usually associated with deciduous hosts. In conclusion, the broad range of mycorrhizal fungi recovered from the roots of P. media suggests that specialization is not a major factor in determining its distribution.     

Biotic interactions with mycorrhizal systems as extended nutrient acquisition strategies shaping forest soil communities and functions

Plant nutrient acquisition strategies involving ectomycorrhizal (EcM) and arbuscular mycorrhizal (AM) associations, are key plant functional traits leading to distinct carbon (C) and nutrient dynamics in forests. Yet, little is known about how these strategies influence the structure and functioning of soil communities, and if such mycorrhizal effects may be more or less pronounced depending on the type of forest and various abiotic factors. Here we explore the potential interactions occurring between plant-EcM and plant-AM systems with the diverse soil organisms occurring in forest soils, and in the process draw attention to major issues that are worthy for future research directions. Based on these potential interactions, we suggest that EcM systems, especially those involving gymnosperms in colder climates, may select for a soil community with a narrow set of functions. These EcM systems may exhibit low functional redundancy, dominated by symbiotic interactions, where EcM fungi maintain low pH and high C/N conditions in order to tightly control nutrient cycling and maintain the dominance of EcM trees. By contrast, AM systems, particularly those involving deciduous angiosperm trees in mild and warmer climates, may facilitate a functionally more diverse and redundant soil community tending towards the dominance of competitive and antagonistic interactions, but also with a range of symbiotic interactions that together maintain diverse plant communities. We propose that the contrasting belowground interactions in AM and EcM systems act as extended nutrient acquisition traits that contribute greatly to the prevailing nutrient and C dynamics occurring in these systems.     

Patterns of Forest Decline and Regeneration Across Scots Pine Populations

To predict future changes in forest ecosystems, it is crucial to understand the complex processes involved in decline of tree species populations and to evaluate the implications for potential vegetation shifts. Here, we study patterns of decline (canopy defoliation and mortality of adults) of four Scots pine populations at the southern edge of its distribution and characterized by different combinations of climate dryness and intensity of past management. General linear and structural equation modeling were used to assess how biotic, abiotic, and management components interacted to explain the spatial variability of Scots pine decline across and within populations. Regeneration patterns of Scots pine and co-occurring oak species were analyzed to assess potential vegetation shifts. Decline trends were related to climatic dryness at the regional scale, but, ultimately, within-population forest structure, local site conditions, and past human legacies could be the main underlying drivers of Scots pine decline. Overall, Scots pine regeneration was negatively related to decline both within and between populations, whereas oak species responded to decline idiosyncratically across populations. Taken together, our results suggest that (1) patterns of decline are the result of processes acting at the plot level that modulate forest responses to local environmental stress and (2) decline of adult Scots pine trees seems not to be compensated by self-recruitment so that the future dynamics of these forest ecosystems are uncertain.     

Presence of arbuscular mycorrhizas in typically ectomycorrhizal host species from Cameroon and New Zealand

 Ectomycorrhizas (EcM) and arbuscular mycorrhizas (AM) were screened for in saplings of 14 EcM tree species from the N'Dupé and Korup National Park rainforests, SW Cameroon, belonging to Caesalpiniaceae and Uapacaceae. The pattern of EcM and AM colonisation of a dual mycorrhizal species from this rainforest (Uapaca staudtii, Uapacaceae) was compared with dual EcM/AM colonisation of Leptospermum scoparium (Myrtaceae) from New Zealand. Both species were collected in a range of habitats. EcM and AM colonisation differed among species in the Korup National Park rainforest: 12 species belonging to the Caesalpiniaceae (Amherstieae) were consistently EcM, and AM structures occurred occasionally in six of them; two other species belonging to Caesalpiniaceae (Afzelia bipindensis) and Uapacaceae (U. staudtii) were dual mycorrhizal with variable levels of colonisation by both EcM and AM fungi. EcM and AM dual colonisation varied with both habitat and identity of the partners. The presence of EcM fungi in most of the root samples of U. staudtii and a negative relationship between AM and EcM colonisation within the same root system suggested a greater EcM affinity of this species. In contrast, most root samples of L. scoparium were colonised by AM, but only a few by EcM. Genuine dual EcM/AM associations in root samples of U. staudtii where the two mycorrhizal types co-occurred could be attributed to an AM-EcM succession. However, differences between predicted and observed frequencies of genuine dual EcM/AM associations in several samples of both U. staudtii and L. scoparium indicated that other factors influenced dual EcM/AM associations. The results of this study showed the importance of the identity of the host species in determining the pattern of dual EcM and AM colonisation. Accepted: 18 September 1998     

Recent 14C-labelled assimilate allocation to Scots pine seedling root and mycorrhizosphere compartments developed on reconstructed podzol humus,E- and B- mineral horizons

In northern boreal forests, podzolic soils prevail that comprise of a distinct upper organic humus/mor (O) horizon that is supported by underlying eluvial (E) and illuvial (B) mineral horizons. The dominant tree species, Scots pine (Pinus sylvestris L.), is known to be highly dependent on root symbiosis with ectomycorrhizal fungi that develop in constituent podzol horizons for growth in these nutrient limited soils. The aim of this microcosm-based study was a quantification of photosynthetically fixed ¹⁴C allocation, following standard pulse-feeding of 7-month-old Scots pine seedling shoots, to respective root and mycorrhizosphere compartments that developed in the reconstructed podzol (O, E and B) profile. Biomass of roots and mycorrhizas decreased with increasing soil depth but no soil origin, control forest vs. clear-cut area, related differences were observed. Similarly, no major soil origin- or podzol horizon-related differences in categorised ectomycorrhizal morphotypes and number of mycorrhizas, in relation to pooled root and mycorrhiza biomass, were detected. However, the total recovery of ¹⁴C-label was significantly higher in clear-cut soil microcosms compared to control counterparts. A significant finding was equivalent ¹⁴C-carbon allocation to roots and ectomycorrhizas in all three major, organic and mineral, podzol profile horizons studied. These carbon allocation data provide additional support for direct (or indirect) roles of roots and symbiotic mycorrhizal fungi in mineral weathering and biodegradation of organic ligands that are central for plant acquisition of growth limiting nutrients and the podzolization process in boreal forest ecosystems.     

Post-logging organic matter recovery in forest ecosystems of eastern Baikal region

The dynamics of organic matter accumulated in the soil and main vegetation elements was analyzed for post-logging forest ecosystem succession series in eastern Baikal region. The phytomass was found to allocate up 63 and 50% of carbon in undisturbed Scots pine and fir stands, respectively. The post-logging phytomass contribution to the total carbon pool appeared to decrease down to 16% in Scots pine and 6% in fir stands. In Scots pine stands, carbon storage was determined to account for almost 70% of the initial carbon 60 years after logging. In 50- to 55-year-old fir stands, carbon recovered its initial pool only by 10%. Soil carbon recorded in recently logged Scots pine and fir sites appeared to be 5 and 16 times that accumulated in the phytomass, respectively. The ratio between phytomass carbon and soil organic matter recovered back to the prelogging level in Scots pine stands by the age of 50–60 years. While phytomass carbon also increased in fir stand of the same age, it did not reach the level of the control stand.     

The effects of heat treatments on ectomycorrhizal resistant propagules and their ability to colonize bioassay seedlings

The effect of disturbance on the resistant propagule community (RPC) of ectomycorrhizal fungi has been given relatively little attention. In this study we investigate the effects of heat, one important factor of fire disturbances, on the ability of ectomycorrhizal RPC fungi to colonize Pinus jeffreyi seedlings in greenhouse bioassays. Prior to planting the seed, soils were collected from an old growth mixed-conifer forest in the southern Sierra Nevada, California, USA and then subjected to four heat treatments of none, 45 °C, 60 °C, and 75 °C. After eight months, seedlings were harvested and the ectomycorrhizal fungi colonizing the roots were characterized by molecular methods (PCR-RFLP and DNA sequencing). Rhizopogon species increased in dominance on seedlings grown in soils receiving the 75 °C heat treatment. One species significantly increased in frequency, Rhizopogon olivaceotinctus, and two species (Cenococcum geophilum and Wilcoxina sp.) significantly decreased in frequency in the 75 °C treatment. The increase of R. olivaceotinctus, coupled with other features of its behavior, suggests that substantial heat disturbances may benefit this species in competing for roots.     

The effects of heat treatment on some technological properties of Scots pine (Pinus sylvestris L.) wood

Heat treatment is often applied to wood species to improve their dimensional stability. This study examined the effect of heat treatment on certain mechanical properties of Scots pine (Pinus sylvestris L.), which has industrially high usage potential and large plantations in Turkey. Wood specimens obtained from Bolu, Turkey, were subjected to heat treatment under atmospheric pressure at varying temperatures (120, 150 and 180 degrees C) for varying durations (2, 6 and 10h). The test results of heat-treated Scots pine and control samples showed that technological properties including compression strength, bending strength, modulus of elasticity in bending, janka-hardness, impact bending strength and tension strength perpendicular to grain suffered with heat treatment, and increase in temperature and duration further diminished technological strength values of the wood specimens.     

Spatial structure and the effects of host and soil environments on communities of ectomycorrhizal fungi in wooded savannas and rain forests of Continental Africa and Madagascar

Mycorrhizal fungi play a key role in mineral nutrition of terrestrial plants, but the factors affecting natural distribution, diversity and community composition of particularly tropical fungi remain poorly understood. This study addresses shifts in community structure and species frequency of ectomycorrhizal (EcM) fungi in relation to host taxa, soil depth and spatial structure in four contrasting African ecosystems. We used the rDNA and plastid trnL intron sequence analysis for identification of fungi and host plants, respectively. By partitioning out spatial autocorrelation in plant and fungal distribution, we suggest that African EcM fungal communities are little structured by soil horizon and host at the plant species and family levels. These findings contrast with patterns of vegetation in these forests and EcM fungal communities in other tropical and temperate ecosystems. The low level of host preference indirectly supports an earlier hypothesis that pioneer Phyllanthaceae may facilitate the establishment of late successional Fabaceae and potentially other EcM host trees by providing compatible fungal inoculum in deforested and naturally disturbed ecosystems of tropical Africa.     

The production of ectomycorrhizal mycelium in forests: Relation between forest nutrient status and local mineral sources

Due to acid rain and nitrogen deposition, there is growing concern that other mineral nutrients, primarily potassium and phosphorus, might limit forest production in boreal forests. Ectomycorrhizal (EcM) fungi are important for the acquisition of potassium and phosphorus by trees. In a field investigation, the effects of poor potassium and phosphorus status of forest trees on the production of EcM mycelium were examined. The production of EcM mycelium was estimated in mesh bags containing sand, which were buried in the soil of forests of different potassium and phosphorus status. Mesh bags with 2% biotite or 1% apatite in sand were also buried to estimate the effect of local sources of nutrients on the production of EcM mycelium. No clear relation could be found between the production of EcM mycelium and nutrient status of the trees. Apatite stimulated the mycelial production, while biotite had no significant effect. EcM root production at the mesh bag surfaces was stimulated by apatite amendment in a forest with poor phosphorus status. The contribution of EcM fungi to apatite weathering was estimated by using rare earth elements (REE) as marker elements. The concentration of REE was 10 times higher in EcM roots, which had grown in contact with the outer surface of apatite-amended mesh bags than in EcM roots grown in contact with the biotite amended or sand-filled mesh bags. In a laboratory study, it was confirmed that REE accumulated in the roots with very low amounts <1 translocated to the shoots. The short-term effect of EcM mycelium on the elemental composition of biotite and apatite was investigated and compared with biotite- and apatite-amended mesh bags buried in trenched soil plots, which were free from EcM fungi. The mesh bags subjected to EcM fungi showed no difference in chemical composition after 17 months in the field. This study suggests that trees respond to phosphorus limitation by increased exploitation of phosphorus-containing minerals by ectomycorrhiza. However, the potential to ameliorate potassium limitation in a similar way appears to be low.     

Ectomycorrhizal fungi diversity in a white sand forest in western Amazonia

The genera Dicymbe and Aldina (Fabaceae) host ectomycorrhizal fungi (EcM) and are common in white sand forests (WSFs), a highly specialized habitat with a high level of plant endemism compared with terra-firme forests. In this study, we visited four times a 1-ha permanent plot established in a small patch of a WSF in the south of Colombia Amazonia. Forty-eight species of EcM fungi were recovered from sporocarps and 15 ITS species-level were detected from root tips. Seventeen species were new reports to Colombia and seven corresponded to undescribed species. These results confirm that this WSF supports a significant EcM fungal diversity. Most of the species found in this study have been previously reported to be associated with other legume and/or dipterocarp species from geographically distant forests. The long-distance occurrence combined with low host specificity, suggest the possibility of gene flow between geographically distant populations of EcM fungi in neotropical lowland rainforests.     

Asynchrony in larval development of the pine beauty moth, Panolis flammea, on an introduced host plant may affect parasitoid efficacy

In the United Kingdom, Panolis flammea (Den. and Schiff.) (Lepidoptera: Noctuidae) is an important pest species of the introduced lodgepole pine but not of its natural host Scots pine. The timing of P. flammea larval growth must be synchronized with its host tree if the larvae are to succeed. We collected field data during 1990 which revealed that the phenological window starts earlier in Scots pine and is shorter than that observed in lodgepole pine. The larvae are found in the field earlier and within a narrower time frame within a Scots pine forest than in a lodgepole pine forest. The larval developmental period is significantly longer on lodgepole pine than on Scots pine. The synchrony/asynchrony of P. flammea to its natural host (Scots pine) and an introduced tree (lodgepole pine) results in the parasitoids having a different impact on the larvae of the two hosts. At any one time, the host plant, caterpillars and parasitoids are more synchronous on the ancestral Scots pine than on lodgepole pine, resulting in a higher percentage of larvae in the optimal instar for parasitism at that time. In lodgepole pine, the percentage of suitable instars available to parasitoids is lower at any given time. The information presented here furthers our understanding of the possible mechanisms for the observed differential population dynamics of the insect on Scots pine and lodgepole pine in the UK. Handling editor: Robert Glinwood.