Ectomycorrhizal Fungal Symbionts Research Articles

Characterization of ectomycorrhizal fungal community of Abies Pindrow using sporocarp sampling, morphotyping, and metabarcoding through next-generation sequencing.

Abies pindrow, commonly known as the West-Himalayan Fir, holds great ecological importance as a native tree species in the Himalayas. Beyond its value as a fuel and timber source, it serves as a keystone species within the ecosystem. However, over recent years, extensive degradation and deforestation have afflicted A. pindrow forests. Utilizing ectomycorrhizal fungal symbionts of A. pindrow could prove pivotal in restoring these deteriorated forests. This study aimed to evaluate the diversity and composition of the ectomycorrhizal fungal community associated with A. pindrow. We employed ectomycorrhizal root tip morphotyping, sporocarp sampling, and Illumina MiSeq metabarcoding of the ITS region of fungal nrDNA. The ectomycorrhizal root tips were categorized into 10 morphotypes based on their morphological characteristics, exhibiting an average colonization rate of 74%. Sporocarp sampling revealed 22 species across 10 genera, with Russula being the most prevalent. The metabarcoding yielded 285,148 raw sequences, identifying 326 operational taxonomic units (OTUs) belonging to 193 genera, 114 families, 45 orders, 22 classes, and 6 divisions. Of these, 36 OTUs across 20 genera were ectomycorrhizal, constituting 63.1% of the fungal community. Notably, Tuber was the most abundant, representing 37.42% of the fungal population, followed by Russula at 21.06%. This study provides a comprehensive understanding of mycorrhizal symbionts of A. pindrow. The findings hold significant implications for utilizing dominant ectomycorrhizal fungi in reforestation endeavors aimed at restoring this important Himalayan conifer.

Restinga ectomycorrhizae: a work in progress.

Background: The Brazilian Atlantic Forest is one of the most biodiverse terrestrial ecoregions of the world. Among its constituents, restinga vegetation makes a particular case, acting as a buffer zone between the oceans and the forest. Covering some 80% of Brazilian coastline (over 7,300 km in length), restinga is a harsh environment where plants and fungi interact in complex ways that just now are beginning to be unveiled. Ectomycorrhizal symbiosis, in particular, plays a so far ungauged and likely underestimated role. We recently described the morpho-anatomical and molecular features of the ectomycorrhizae formed by several basidiomycetous mycobionts on the host plant Guapira opposita, but the mycorrhizal biology of restinga is still largely unexplored. Here, we report new data on the ectomycorrhizal fungal symbionts of G. opposita, based on the collection of sporomata and ectomycorrhizal root tips in restinga stands occurring in southern Brazil. Methods: To obtain a broader view of restinga mycorrhizal and ecological potential, we compiled a comprehensive and up-to-date checklist of fungal species reported or supposed to establish ectomycorrhizae on restinga-inhabiting host plants, mainly on the basis of field observations. Results: Our list comprises some 726 records, 74 of which correspond to putative ectomycorrhizal taxa specifically associated with restinga. These include several members of Boletaceae, Amanita, Tomentella/ Thelephora, Russula/ Lactifluus, and Clavulina, as well as hypogeous fungi, like the recently described Longistriata flava. Conclusions: Our survey reveals a significant diversity of the restinga ectomycorrhizal mycobiota, indicating the importance of this symbiosis for the ecological functioning of a unique yet poorly known and threatened ecosystem.

Traits and tradeoffs among non-native ectomycorrhizal fungal symbionts affect pine seedling establishment in a Hawaiian coinvasion landscape.

Pine invasions lead to losses of native biodiversity and ecosystem function, but pine invasion success is often linked to coinvading non-native ectomycorrhizal (EM) fungi. How the community composition, traits, and distributions of these fungi vary over the landscape and how this affects pine success is understudied. A greenhouse bioassay experiment was performed to test the effects of changes in EM fungal community structure from a pine plantation, to an invasion front to currently pine-free areas on percent root colonization and seedling biomass. Soils were also analysed by qPCR to determine changes in inoculum and spore density over distance for a common coinvading EM fungus, Suillus pungens. Percent colonization increased with distance from the plantation, which corresponded with an increase in seedling biomass and stark changes in EM fungal community membership where Suillus spp. dominated currently pine-free areas. However, there was a negative relationship between S.pungens inoculum potential versus root colonization over distance. We conclude that the success of pine invasions is facilitated by specific traits of Suillus spp., but that the success of Suillus is contingent on a lack of competition with other ectomycorrhizal fungi.

Phylogenetic diversity of 200+ isolates of the ectomycorrhizal fungus Cenococcum geophilum associated with Populus trichocarpa soils in the Pacific Northwest, USA and comparison to globally distributed representatives.

The ectomycorrhizal fungal symbiont Cenococcum geophilum is of high interest as it is globally distributed, associates with many plant species, and has resistance to multiple environmental stressors. C. geophilum is only known from asexual states but is often considered a cryptic species complex, since extreme phylogenetic divergence is often observed within nearly morphologically identical strains. Alternatively, C. geophilum may represent a highly diverse single species, which would suggest cryptic but frequent recombination. Here we describe a new isolate collection of 229 C. geophilum isolates from soils under Populus trichocarpa at 123 collection sites spanning a ~283 mile north-south transect in Western Washington and Oregon, USA (PNW). To further understanding of the phylogenetic relationships within C. geophilum, we performed maximum likelihood and Bayesian phylogenetic analyses to assess divergence within the PNW isolate collection, as well as a global phylogenetic analysis of 789 isolates with publicly available data from the United States, Japan, and European countries. Phylogenetic analyses of the PNW isolates revealed three distinct phylogenetic groups, with 15 clades that strongly resolved at >80% bootstrap support based on a GAPDH phylogeny and one clade segregating strongly in two principle component analyses. The abundance and representation of PNW isolate clades varied greatly across the North-South range, including a monophyletic group of isolates that spanned nearly the entire gradient at ~250 miles. A direct comparison between the GAPDH and ITS rRNA gene region phylogenies, combined with additional analyses revealed stark incongruence between the ITS and GAPDH gene regions, consistent with intra-species recombination between PNW isolates. In the global isolate collection phylogeny, 34 clades were strongly resolved using Maximum Likelihood and Bayesian approaches (at >80% MLBS and >0.90 BPP respectively), with some clades having intra- and intercontinental distributions. Together these data are highly suggestive of divergence within multiple cryptic species, however additional analyses such as higher resolution genotype-by-sequencing approaches are needed to distinguish potential species boundaries and the mode and tempo of recombination patterns.

Halimium as an ectomycorrhizal symbiont: new records and an appreciation of known fungal diversity

Halimium is a genus of Cistaceae, containing a small group of shrub species found in open vegetation types and in degraded forest patches throughout the western and central Mediterranean region. We recently described the morpho-anatomical features of the ectomycorrhizae formed by Scleroderma meridionale on Halimium halimifolium, but the mycorrhizal biology of this host plant genus is still largely unexplored. Here, we report new data on the ectomycorrhizal fungal symbionts of Halimium, based on the collection of sporocarps and ectomycorrhizal root tips in pure stands occurring in Sardinia, Italy. To obtain a broader view of Halimium mycorrhizal and ecological potential, we compiled a comprehensive and up-to-date checklist of fungal species reported to establish ectomycorrhizae on Halimium spp. on the basis of field observations, molecular approaches, and mycorrhiza synthesis. Our list comprises 154 records, corresponding to 102 fungal species and 35 genera, revealing a significant diversity of the Halimium ectomycorrhizal mycobiota. Key ectomycorrhizal genera like Russula, Lactarius/Lactifluus, Amanita, Inocybe, and Cortinarius account for more than half of all mycobionts. A large proportion of Halimium fungal species are shared with other host plants in various ecological settings, suggesting a critical role of common mycorrhizal networks in the function played by this shrub in various Mediterranean ecosystems.

Protein Arginine Methyltransferase Expression Affects Ectomycorrhizal Symbiosis and the Regulation of Hormone Signaling Pathways.

The genomes of all eukaryotic organisms, from small unicellular yeasts to humans, include members of the protein arginine methyltransferase (PRMT) family. These enzymes affect gene transcription, cellular signaling, and function through the posttranslational methylation of arginine residues. Mis-regulation of PRMTs results in serious developmental defects, disease, or death, illustrating the importance of these enzymes to cellular processes. Plant genomes encode almost the full complement of PRMTs found in other higher organisms, plus an additional PRMT found uniquely in plants, PRMT10. Here, we investigate the role of these highly conserved PRMTs in a process that is unique to perennial plants-the development of symbiosis with ectomycorrhizal fungi. We show that PRMT expression and arginine methylation is altered in the roots of the model tree Eucalyptus grandis by the presence of its ectomycorrhizal fungal symbiont Pisolithus albus. Further, using transgenic modifications, we demonstrate that E. grandis-encoded PRMT1 and PRMT10 have important but opposing effects in promoting this symbiosis. In particular, the plant-specific EgPRMT10 has a potential role in the expression of plant hormone pathways during the colonization process and its overexpression reduces fungal colonization success.

Fenton reaction facilitates organic nitrogen acquisition by an ectomycorrhizal fungus.

Summary Boreal trees rely on their ectomycorrhizal fungal symbionts to acquire growth‐limiting nutrients, such as nitrogen (N), which mainly occurs as proteins complexed in soil organic matter (SOM). The mechanisms for liberating this N are unclear as ectomycorrhizal fungi have lost many genes encoding lignocellulose‐degrading enzymes present in their saprotrophic ancestors. We hypothesized that hydroxyl radicals (˙ OH), produced by the ectomycorrhizal fungus Paxillus involutus during growth on SOM, are involved in liberating organic N. Paxillus involutus was grown for 7 d on N‐containing or N‐free substrates that represent major organic compounds of SOM. ˙ OH production, ammonium assimilation, and proteolytic activity were measured daily. ˙ OH production was strongly induced when P. involutus switched from ammonium to protein as the main N source. Extracellular proteolytic activity was initiated shortly after the oxidation. Oxidized protein substrates induced higher proteolytic activity than unmodified proteins. Dynamic modeling predicted that ˙ OH production occurs in a burst, regulated mainly by ammonium and ferric iron concentrations.We propose that the production of ˙ OH and extracellular proteolytic enzymes are regulated by similar nutritional signals. Oxidation works in concert with proteolysis, improving N liberation from proteins in SOM. Organic N mining by ectomycorrhizal fungi has, until now, only been attributed to proteolysis.

Ectomycorrhizal fungal assemblages of Abies alba Mill. outside its native range in Poland.

Abies alba (Mill.) is an important forest tree species, native to the mountainous regions of Europe but has been also widely introduced in the lowlands outside its native range. Like most forest tree species, A. alba forms obligate mutualisms with ectomycorrhizal (ECM) fungi. This investigation sought to examine ECM fungal communities of A. alba when the species grows 400 km north of its native range in the region of Pomerania in Poland. We surveyed for ECM fungi by sampling live roots from four mature forest stands where the A. alba component ranged from 20 to 100 %. Ectomycorrhizal fungal symbionts were identified based on morphotyping and sequencing of the internal transcribed spacer (ITS) of nuclear ribosomal DNA (rDNA). Thirty-five ECM fungal taxa were distinguished on root tips of A. alba from all tested stands with 22 to 27 ECM fungal taxa in the individual stand. The diversity and similarity metrics revealed a lack of statistical differences in the structure of the ECM fungal community between stands varying in overstory tree composition. Cenococcum geophilum was the most common fungal species at all investigated A. alba stands, with an abundance of 50 to 70 %. The ECM community was characterized by the lack of Abies-specific fungal symbionts and a rich and diverse suite of host-generalist mycobionts that seem to be sufficient for successful growth and development of A. alba outside of its native range.

A single ectomycorrhizal fungal species can enable a Pinus invasion.

Like all obligately ectomycorrhizal plants, pines require ectomycorrhizal fungal symbionts to complete their life cycle. Pines introduced into regions far from their native range are typically incompatible with local ectomycorrhizal fungi, and, when they invade, coinvade with fungi from their native range. While the identities and distributions of coinvasive fungal symbionts of pine invasions are poorly known, communities that have been studied are notably depauperate. However, it is not yet clear whether any number of fungal coinvaders is able to support a Pinaceae invasion, or whether very depauperate communities are unable to invade. Here, we ask whether there is evidence for a minimum species richness of fungal symbionts necessary to support a pine/ectomycorrhizal fungus coinvasion. We sampled a Pinus contorta invasion front near Coyhaique, Chile, using molecular barcoding to identify ectomycorrhizal fungi. We report that the site has a total richness of four species, and that many invasive trees appear to be supported by only a single ectomycorrhizal fungus, Suillus luteus. We conclude that a single ectomycorrhizal (ECM) fungus can suffice to enable a pine invasion.

Diversity and spatial structure of belowground plant-fungal symbiosis in a mixed subtropical forest of ectomycorrhizal and arbuscular mycorrhizal plants.

Plant–mycorrhizal fungal interactions are ubiquitous in forest ecosystems. While ectomycorrhizal plants and their fungi generally dominate temperate forests, arbuscular mycorrhizal symbiosis is common in the tropics. In subtropical regions, however, ectomycorrhizal and arbuscular mycorrhizal plants co-occur at comparable abundances in single forests, presumably generating complex community structures of root-associated fungi. To reveal root-associated fungal community structure in a mixed forest of ectomycorrhizal and arbuscular mycorrhizal plants, we conducted a massively-parallel pyrosequencing analysis, targeting fungi in the roots of 36 plant species that co-occur in a subtropical forest. In total, 580 fungal operational taxonomic units were detected, of which 132 and 58 were probably ectomycorrhizal and arbuscular mycorrhizal, respectively. As expected, the composition of fungal symbionts differed between fagaceous (ectomycorrhizal) and non-fagaceous (possibly arbuscular mycorrhizal) plants. However, non-fagaceous plants were associated with not only arbuscular mycorrhizal fungi but also several clades of ectomycorrhizal (e.g., Russula) and root-endophytic ascomycete fungi. Many of the ectomycorrhizal and root-endophytic fungi were detected from both fagaceous and non-fagaceous plants in the community. Interestingly, ectomycorrhizal and arbuscular mycorrhizal fungi were concurrently detected from tiny root fragments of non-fagaceous plants. The plant–fungal associations in the forest were spatially structured, and non-fagaceous plant roots hosted ectomycorrhizal fungi more often in the proximity of ectomycorrhizal plant roots. Overall, this study suggests that belowground plant–fungal symbiosis in subtropical forests is complex in that it includes “non-typical” plant–fungal combinations (e.g., ectomycorrhizal fungi on possibly arbuscular mycorrhizal plants) that do not fall within the conventional classification of mycorrhizal symbioses, and in that associations with multiple functional (or phylogenetic) groups of fungi are ubiquitous among plants. Moreover, ectomycorrhizal fungal symbionts of fagaceous plants may “invade” the roots of neighboring non-fagaceous plants, potentially influencing the interactions between non-fagaceous plants and their arbuscular-mycorrhizal fungal symbionts at a fine spatial scale.

Testing the link between community structure and function for ectomycorrhizal fungi involved in a global tripartite symbiosis

Alnus trees associate with ectomycorrhizal (ECM) fungi and nitrogen-fixing Frankia bacteria and, although their ECM fungal communities are uncommonly host specific and species poor, it is unclear whether the functioning of Alnus ECM fungal symbionts differs from that of other ECM hosts. We used exoenzyme root tip assays and molecular identification to test whether ECM fungi on Alnus rubra differed in their ability to access organic phosphorus (P) and nitrogen (N) when compared with ECM fungi on the non-Frankia host Pseudotsuga menziesii. At the community level, potential acid phosphatase (AP) activity of ECM fungal root tips from A.rubra was significantly higher than that from P.menziesii, whereas potential leucine aminopeptidase (LA) activity was significantly lower for A.rubra root tips at one of the two sites. At the individual species level, there was no clear relationship between ECM fungal relative root tip abundance and relative AP or LA enzyme activities on either host. Our results are consistent with the hypothesis that ECM fungal communities associated with Alnus trees have enhanced organic P acquisition abilities relative to non-Frankia ECM hosts. This shift, in combination with the chemical conditions present in Alnus forest soils, may drive the atypical structure of Alnus ECM fungal communities.

Revisiting the host effect on ectomycorrhizal fungal communities: implications from host–fungal associations in relict Pseudotsuga japonica forests

Host identity is among the most important factors in structuring ectomycorrhizal (ECM) fungal communities. Both host-fungal coevolution and host shifts can account for the observed host effect, but their relative significance in ECM fungal communities is not well understood. To investigate these two host-related mechanisms, we used relict forests of Pseudotsuga japonica, which is an endangered endemic species in Japan. As with other Asian Pseudotsuga species, P. japonica has been isolated from North American Pseudotsuga spp. since the Oligocene and has evolved independently as a warm-temperate species. We collected 100 soil samples from four major localities in which P. japonica was mixed with other conifers and broadleaf trees. ECM tips in the soil samples were subjected to molecular analyses to identify both ECM fungi and host species. While 136 ECM fungal species were identified in total, their communities were significantly different between host groups, confirming the existence of the host effect on ECM fungal communities. None of the 68 ECM fungal species found on P. japonica belonged to Pseudotsuga-specific lineages (e.g., Rhizopogon and Suillus subgroups) that are common in North America. Most of ECM fungi on P. japonica were shared with other host fungi or phylogenetically close to known ECM fungi on other hosts in Asia. These results suggest that after migrating, Pseudotsuga-specific fungal lineages may have become extinct in small isolated populations in Japan. Instead, most of the ECM fungal symbionts on P. japonica likely originated from host shifts in the region.

Identities and distributions of the co-invading ectomycorrhizal fungal symbionts of exotic pines in the Hawaiian Islands

Pine species have become invasive throughout the globe and threaten to replace native biota. The threat of pine invasion is particularly pressing in parts of the tropics where there are no native pines. The factors that govern pine invasion are not often well understood. However, key to pine survival is an obligate and mutualistic interaction with ectomycorrhizal fungi. Thus for pines to successfully invade new habitats compatible ectomycorrhizal fungi must already be present, or be co-introduced. The purpose of this study was to examine the community structure of non-native ectomycorrhizal fungi associated with pine invasions in the Hawaiian Islands. To accomplish this we executed a field and greenhouse study and used a molecular ecology approach to identify the fungi associating with invasive pines in Hawai‘i. We show that: (1) ectomycorrhizal fungal species richness in non-native pine plantations is far less than what is found in pine’s native range, (2) there was a significant decrease in average ectomycorrhizal fungal species richness as distance from pine plantations increased and, (3) Suillus species were the dominant fungi colonizing pines outside plantations. The keystone ectomycorrhizal fungal taxa responsible for pine establishment in Hawai‘i are within genera commonly associated with pine invasions throughout the globe. We surmise that these fungi share functional traits such as the ability for long-distance dispersal from plantations and host tree colonization via spore that lead to their success when introduced to new habitats.

Coarse woody debris retention in subalpine clearcuts affects ectomycorrhizal root tip community structure within fifteen years of harvest

Shifts in ectomycorrhizal (ECM) fungal community composition occur after clearcut logging, resulting in the loss of some dominant forest fungi. Because decaying wood is a remnant of the original forest and an important habitat for ECM fungal species, we examined ECM spruce roots and hyphae in 1-ha coarse woody debris (CWD) retention and removal plots (N=3) at a high elevation spruce forest 12 and 13 years after clearcut harvesting to test for a medium-term effect of CWD retention. Root tips from ten Picea engelmannii (Parry ex. Engelm.) saplings per plot were grouped morphologically, and the ECM fungal symbiont identified by Sanger sequencing. Sand-filled hyphae-trapping mesh bags were buried amongst the roots of saplings for one year. PCR product from all bags per plot was pooled and submitted for GS-FLX Titanium sequencing. Forty-six of 89 operational taxonomic units (OTUs) from root tips and 50 OTUs from hyphae were identified as ECM taxa. The most abundant taxa amongst root tip OTUs were Thelephora terrestris, Alloclavaria purpurea, Amphinema byssoides, and Tylospora asterophora. The most abundant OTUs from mesh bags were ectomycorrhizal taxa, and the most abundant of these were A. byssoides, T. terrestris, Wilcoxina mikolae, and T. asterophora. The retention of CWD had no detectable effect on taxon richness, evenness or diversity of ECM fungi on root tips or in mesh bags; however, there was a detectable shift in species composition. The relative abundance of A. byssoides root tips was significantly higher at removal plots while A. purpurea was a significant indicator species for CWD removal. Our results suggest that the retention of CWD at the time of harvest has affected ECM habitat at this site, and has resulted in altered ECM species composition, even though the logs are still hard and intact.

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.

Ectomycorrhizal identity determines respiration and concentrations of nitrogen and non-structural carbohydrates in root tips: a test using Pinus sylvestris and Quercus robur saplings

Fine roots play a significant role in plant and ecosystem respiration (RS); therefore, understanding factors controlling that process is important both to advancing understanding and potentially in modelling carbon (C) budgets. However, very little is known about the extent to which ectomycorrhizal (ECM) identity may influence RS or the underlying chemistry that may determine those rates. In order to test these relationships, we examined RS, measured as O(2) consumption, of first-order ECM root tips of Pinus sylvestris L. and Quercus robur L. saplings in relation to their ECM fungal symbionts and associated nitrogen (N), C and non-structural carbohydrate concentrations. Roots of P. sylvestris were colonized by Rhizopogon roseolus, Tuber sp. 1 and an unknown species of Pezizales. Fungal species colonizing Q. robur roots were Hebeloma sp., Tuber sp. 2 and one unidentified ECM fungus described as Tuber-like based on ECM morphology. ECM RS rates for different host species were significantly different and more than 97% of the variation in RS within a host species was explained by ECM root tip N concentrations. This may indicate that some of the variability in fine root RS-N relationships observed between and within different host species or their functional groups may be related to intraspecific host species differences in root tip N concentration among ECM fungal associates.

Paxillus involutus Forms an Ectomycorrhizal Symbiosis and Enhances Survival of PtCOMT-modified Betula pendula in vitro

AbstractThe ability of the PtCOMT (caffeate/5-hydroxyferulate O-methyltransferase from Populus tremuloides L.) - modified Betula pendula Roth. lines to form symbiosis with an ectomycorrhizal (ECM) fungus Paxillus involutus Batsch Fr. was studied in vitro. Lignin precursor gene PtCOMT was introduced into two B. pendula clones under the control of the cauliflower mosaic virus 35S promoter or the promoter of the sunflower polyubiquitin gene UbB1. Of the four transgenic lines, one 35SPtCOMT line (23) had a decreased syringyl/guaiacyl (S/G) ratio of root lignin, and two UbB1-PtCOMT lines (110 and 130) retarded root growth compared to the control clone. Both control clones and all transgenic lines were able to form ECMs with P. involutus, but the transgenic lines differed from the controls in the characteristics of the ECMs. The number of lateral roots covered with fungal hyphae and/or development of a Hartig net (HN) were reduced in line 23 with a decreased S/G ratio, and in lines 110 and 130 with slower root formation and changed root morphology, respectively. However, line 23 benefited more from the inoculation in lateral root formation than the control, and in lines 110 and 130 the percentage of viable plants increased most due to inoculation. The results show that B. pendula plants genetically transformed with the lignin gene PtCOMT could form mycorrhizal symbiosis regardless of changes in either the root S/G ratio or development. The benefits of the symbiosis were variable even in the closed in vitro system, and dependent on the clone or transgenic line and the ECM fungal symbiont.

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.

Effect of soil compaction and moisture on incidence of phytophthora root rot on American chestnut ( Castanea dentata) seedlings

American chestnut is one of hundreds of plant species plagued by root rot caused by Phytophthora cinnamomi. Phytophthora root rot is thought to have contributed to chestnut dieback prior to the arrival of chestnut blight, and it may now present a serious limitation to establishment of blight-resistant hybrid chestnut. We manipulated soil compaction and moisture to evaluate the effect of soil physical factors on incidence of Phytophthora root rot on American chestnut seedlings. Seedlings were grown under three watering regimes, two soil compaction levels and two fungicide levels. Increased soil moisture enhanced seedling growth in loose soil, but irrigation did not impact seedlings growing in compacted soils. Seedling mortality was greatest in wet, compacted soils. Disease incidence was highest in the wettest soils, irrespective of compaction level. Fine root necrosis and Phytophthora infection occurred on 58 and 24% of non-fungicide-treated seedlings, respectively. Presence of ectomycorrhizal fungi declined in compacted soils that were either wetter or drier than optimal. Occurrence of ectomycorrhizal fungal symbionts was unrelated to root rot. This study demonstrates the high susceptibility of American chestnut to this common root pathogen, even under moderate levels of soil compaction and moisture. While overcoming chestnut blight is the first step in restoring chestnut to its original range, to establish successful plantings it will be crucial to recognize and avoid sites where soil physical factors promote Phytophthora root rot.

Influence of ectomycorrhizal fungal inoculation on growth and root IAA concentrations of transplanted conifers.

We determined whether in vitro plant growth regulator production by mycorrhizal fungi is correlated with conifer seedling growth and root IAA concentrations. Container-grown seedlings of interior Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco), lodgepole pine (Pinus contorta Dougl.) and ponderosa pine (Pinus ponderosa Dougl.) were inoculated at seeding with ectomycorrhizal fungi having a high, moderate or low capacity to produce either IAA or ethylene in vitro. Inoculated seedlings were grown for one season in the nursery, harvested in December, cold stored over winter and then transplanted to either a nursery field or a forest site in the spring. Seedling morphology and endogenous IAA in roots were measured immediately after cold storage and again six and 12 months after transplanting. Morphological responses to inoculation varied among different mycorrhizal fungi. Free IAA concentration of roots was increased in some inoculation treatments for all conifer species. In seedlings transplanted to a nursery field, in vitro ethylene-producing capacity of the ectomycorrhizal fungi was highly correlated with more morphological features than in vitro IAA-producing capacity. Both IAA- and ethylene-producing capacity were significantly correlated with more morphological features in seedlings transplanted to a forest site than in seedings transplanted to a nursery field. One year after transplanting, only in vitro IAA-producing capacity was correlated with endogenous IAA concentration of roots of the inoculated seedlings. We conclude that growth responses of conifer seedlings can be partially influenced by IAA and ethylene produced by ectomycorrhizal fungal symbionts.