Melanie D. Jones

Net transfer of carbon between ectomycorrhizal tree species in the field

Different plant species can be compatible with the same species of mycorrhizal fungi, and be connected to one another by a common mycelium,. Transfer of carbon, nitrogen, and phosphorus, through interconnecting mycelia has been measured frequently in laboratory experiments, but it is not known whether transfer is bidirectional, whether there is a net gain by one plant over its connected partner, or whether transfer affects plant performance in the field,. Laboratory studies using isotope tracers show that the magnitude of one-way transfer can be influenced by shading of ‘receiver’ plants,, fertilization of ‘donor’ plants with phosphorus, or use of nitrogen-fixing donor plants and non-nitrogen-fixing receiver plants,, indicating that movement may be governed by source–sink relationships. Here we use reciprocal isotope labelling in the field to demonstrate bidirectional carbon transfer between the ectomycorrhizal tree species Betula papyrifera and Pseudotsuga menziesii, resulting in net carbon gain by P. menziesii. Thuja plicata seedlings lacking ectomycorrhizae absorb small amounts of isotope, suggesting that carbon transfer between B. papyrifera and P. menziesii is primarily through the direct hyphal pathway. Net gain by P. menziesii seedlings represents on average 6% of carbon isotope uptake through photosynthesis. The magnitude of net transfer is influenced by shading of P. menziesii, indicating that source–sink relationships regulate such carbon transfer under field conditions.

Carbon and Nutrient Fluxes Within and Between Mycorrhizal Plants

Mycorrhizal fungi are involved in the uptake of nutrients in exchange for C from host plants, and possibly in the transfer of C and nutrients between plants. Ecto-mycorrhizal fungi (EMF) increase uptake rates of nutrients by a variety of mechanisms, including increased physical access to soil, changes to mycorrhizosphere or hyphosphere chemistry, and alteration of the bacterial community in the mycorrhizosphere. They influence mycorrhizosphere chemistry through release of organic acids and production of enzymes. Movement of nutrients within an ecto-mycorrhizal (EM) mycelial network, as well as exchange of C and nutrients between symbionts, appear to be regulated by source-sink relationships. Estimates of the quantity of plant C partitioned belowground (to roots and EMF) varies widely (40–73%) depending on the methodology used and ecosystem studied, and is affected by several factors such as the identity of plant and fungal species, plant nutrient content, and EM age.

THE MUTUALISM–PARASITISM CONTINUUM IN ECTOMYCORRHIZAS: A QUANTITATIVE ASSESSMENT USING META‐ANALYSIS

Context dependency is deemed to position the outcomes of species interactions along a continuum of mutualism to parasitism. Thus, it is imperative to understand which factors determine where a particular interspecific interaction falls along the continuum. Over the past 20 years research on the ectomycorrhizal symbiosis has resulted in sufficient independent studies to now generalize about the factors and mechanisms that affect host response to ectomycorrhizas. Using meta-analysis we quantitatively evaluated the role of biotic (partner identity and colonization levels of ectomycorrhizal fungi) and abiotic (phosphorus levels) factors in determining host biomass, height, and shoot:root responses to ectomycorrhizal associations. On average, seedlings across multiple host genera increased in total biomass when inoculated with ectomycorrhizal fungi regardless of the identity of the fungal associate; host genera differed in the magnitude of response for both total biomass and shoot:root ratio. Association with different fungal genera modified only host allocation of biomass to shoots and roots. Neither level of colonization on inoculated seedlings nor the level of contamination on control seedlings relative to colonization levels by target fungi on inoculated seedlings was important in explaining variation in effect sizes for any growth response. None of our proposed factors (identity of partners, colonization level, magnitude of contamination, or duration of association) explained variation in effect sizes for shoot height, although in general seedlings were taller when inoculated with ectomycorrhizal fungi. Phosphorus additions did not influence effect sizes. Although the general trend across studies was for a positive response of hosts to ectomycorrhizal inoculation, publication bias and methodological issues effectively reduce and distort the spectrum on which we evaluate host responses to ectomycorrhizal inoculation. Our results indicate that the variation in ectomycorrhizal fungi perceived by the host may be of a discrete (presence/absence of ectomycorrhizal fungi) rather than continuous nature (variation in identity or abundance of ectomycorrhizal fungi).

Carbon allocation and carbon transfer between t Betula papyrifera and t Pseudotsuga menziesii seedlings using a 13C pulse-labeling method

Here we describe a simple method for pulse-labeling tree seedlings with 13CO2(gas), and then apply the method in two related experiments: t (i) comparison of carbon allocation patterns between t Betula papyrifera Marsh. and t Pseudotsuga menziesii (Mirb.) Franco, and t (ii) measurement of one-way belowground carbon transfer from t B. papyrifera to t P. menziesii. Intraspecific carbon allocation patterns and interspecific carbon transfer both influence resource allocation, and consequently development, in mixed communities of t B. papyrifera and t P. menziesii.In preparation for the two experiments, we first identified the appropriate 13CO2(gas) pulse-chase regime for labeling seedlings: a range of pulse (100-mL and 200-mL 99 atom%13 CO2(gas)) and chase (0, 3 and 6 d) treatments were applied to one year-old t B. papyrifera and t P. menziesii seedlings. The amount of 13CO2 fixed immediately after 1.5 h exposure was greatest for both t B. papyrifera (40.8 mg excess 13C) and t P. menziesii (22.9 mg excess 13C) with the 200-mL pulse, but higher 13C loss and high sample variability resulted in little difference in excess13 C content between pulse treatments after 3 d for either species. The average excess 13C root/shoot ratio of t B. papyrifera and t P. menziesii changed from 0.00 immediately following the pulse to 0.61 and 0.87 three and six days later, which reflected translocation of 75% of fixed isotope out of foliage within 3 d following the pulse and continued enrichment in fine roots over 6 d. Based on these results, the 100-mL CO2(gas) and 6-d chase were considered appropriate for the carbon allocation and belowground transfer experiments.In the carbon allocation experiment, we found after 6 d that t B. papyrifera allocated 49% (average 9.5 mg) and t P. menziesii 41% (average 5.8 mg) of fixed isotope to roots, of which over 55% occurred in fine roots in both species. Species differences in isotope allocation patterns paralleled differences in tissue biomass distribution. The greater pulse labeling efficiency of t B. papyrifera compared to t P. menziesii was associated with its two-fold and 13- fold greater leaf and whole seedling net photosynthetic rates, respectively, 53% greater biomass, and 35% greater root/shoot ratio.For the carbon transfer experiment, t B. papyrifera and t P. menziesii were grown together in laboratory rootboxes, with their roots intimately mingled. A pulse of 100 mL13 CO2(gas) was applied to paper birch and one-way transfer to neighboring t P. menziesii was measured after 6 d. Of the excess 13C fixed by t B. papyrifera, 4.7% was transferred to neighboring t P. menziesii, which distributed the isotope evenly between roots and shoots. Of the isotope received by t P. menziesii, we estimated that 93% was taken up through belowground pathways, and the remaining 7% taken up by foliage as13 CO2(gas) respired by t B. papyrifera shoots. These two experiments indicate that t B. papyrifera fixes more total carbon and allocates a greater proportion to its root system than does t P. menziesii, giving it a competitive edge in resource gathering; however, below-ground carbon sharing is of sufficient magnitude that it may help ensure co-existence of the two species in mixed communities.

Ectomycorrhizal fungi contribute to soil organic matter cycling in sub-boreal forests

Soils of northern temperate and boreal forests represent a large terrestrial carbon (C) sink. The fate of this C under elevated atmospheric CO2 and climate change is still uncertain. A fundamental knowledge gap is the extent to which ectomycorrhizal fungi (EMF) and saprotrophic fungi contribute to C cycling in the systems by soil organic matter (SOM) decomposition. In this study, we used a novel approach to generate and compare enzymatically active EMF hyphae-dominated and saprotrophic hyphae-enriched communities under field conditions. Fermentation-humus (FH)-filled mesh bags, surrounded by a sand barrier, effectively trapped EMF hyphae with a community structure comparable to that found in the surrounding FH layer, at both trophic and taxonomic levels. In contrast, over half the sequences from mesh bags with no sand barrier were identified as belonging to saprotrophic fungi. The EMF hyphae-dominated systems exhibited levels of hydrolytic and oxidative enzyme activities that were comparable to or higher than saprotroph-enriched systems. The enzymes assayed included those associated with both labile and recalcitrant SOM degradation. Our study shows that EMF hyphae are likely important contributors to current SOM turnover in sub-boreal systems. Our results also suggest that any increased EMF biomass that might result from higher below-ground C allocation by trees would not suppress C fluxes from sub-boreal soils.

Influence of soil nutrients on ectomycorrhizal communities in a chronosequence of mixed temperate forests

Many factors associated with forests are collectively responsible for controlling ectomycorrhizal (ECM) fungal community structure, including plant species composition, forest structure, stand age, and soil nutrients. The objective of this study was to examine relationships among ECM fungal community measures, local soil nutrients, and stand age along a chronosequence of mixed forest stands that were similar in vegetation composition and site quality. Six combinations of age class (5-, 26-, 65-, and 100-year-old) and stand initiation type (wildfire and clearcut) were replicated on four sites, each representing critical seral stages of stand development in Interior Cedar-Hemlock (ICH) forests of southern British Columbia. We found significant relationships between ECM fungal diversity and both available and organic P; available P was also positively correlated with the abundance of two ECM taxa (Rhizopogon vinicolor group and Cenoccocum geophilum). By contrast, ECM fungal diversity varied unpredictably with total and mineralizable N or C to N ratio. We also found that soil C, N, available P, and forest floor depth did not exhibit strong patterns across stand ages. Overall, ECM fungal community structure was more strongly influenced by stand age than specific soil nutrients, but better correlations with soil nutrients may occur at broader spatial scales covering a wider range of site qualities.

Pathways for below-ground carbon transfer between paper birch and Douglas-fir seedlings

Background: Carbon can move below ground between ectomycorrhizal plants, but the relative importance of transfer through common mycorrhizal networks (CMNs) or soil pathways remains unclear. We studied carbon transfer between paper birch (Betula papyrifera) and Douglas-fir (Pseudotsuga menziesii) seedlings grown in adjacent root-restrictive pouches inside root chambers. Aims: The objective of this study was to compare transfer between CMN pathways and soil pathways by testing if: (1) carbon transfer between paper birch and Douglas-fir is bi-directional, (2) there is a net gain in carbon by one of the tree species; and (3) more carbon is transferred through the CMN pathway than the soil pathway. Methods: Following 8 months in the greenhouse, hyphal linkages crossing root pouches were either severed or left intact. Seedlings were then reciprocally labelled with 13CO2 and 14CO2. Results: We found carbon was transferred bi-directionally, with a 2–3% net gain in carbon by Douglas-fir from paper birch. Both bi-directional and net transfer occurred where the CMN was severed, but transfer was greater where it was left intact, indicating that carbon was transferred through both the soil and CMN pathways. Conclusions: Our results suggest that significant amounts of labelled carbon were transferred between plant species, showing for the first time in a balanced pulse-labelling experiment that approximately three times as much carbon was transferred through CMN pathways than soil pathways. That most carbon transfer occurred through CMNs could affect competitive interactions between establishing seedlings, favouring some included in the CMN while disadvantaging others, thus fundamentally altering our understanding of how interspecific interactions alter community structure.

Ectomycorrhizal hyphae structure components of the soil bacterial community for decreased phosphatase production

Ectomycorrhizal fungi (EMF) provide nutrients to their hosts by means of hyphae that extend beyond nutrient-depleted rhizosphere soil. Soil bacteria may compete with EMF for nutrients or may act synergistically to enhance nutrient supply to hosts. To assess the interactions between hyphae and bacteria, two types of small, sand-filled mesh bags were incubated in a Pseudotsuga menziesii/Betula papyrifera forest. The bags allowed ingrowth by EMF (35-μm mesh) or excluded hyphae (0.5-μm mesh), while allowing migration of soil bacteria. After incubation, bacteria were isolated from bags using a method to enrich for Gram-positive bacteria. Isolates were assayed for phosphatase and N-acetyl glucosaminidase (NAGase) activities to assess the potential to access organic phosphorus and nitrogen. The average phosphatase activities were higher in exclusion than ingrowth bags, while NAGase activities did not differ. Streptomyces isolates, which are expected to be strong competitors and antagonists of EMF, were more prevalent in ingrowth bags and yet had lower phosphatase activities. Furthermore, there were no indications of antagonism between fungi and Streptomyces, as there were no increases in NAGase activities in ingrowth bags. We conclude that fungal hyphae can structure components of the soil bacterial community for decreased extracellular enzyme production.

Ectomycorrhizal colonization and intraspecific variation in growth responses of lodgepole pine

Across different host plant species, the effects of mycorrhizal colonization on host growth parameters can vary, but intraspecific variation in this relationship has rarely been measured. We tested the direction and consistency of the relationship between ectomycorrhizal colonization level and growth responses across seed families of Pinus contorta var. latifolia. Root tips of seedlings from eight full sib seed families varied in levels of ectomycorrhizal fungal colonization from 39% to 100%. We observed positive, negative, or neutral relationships between colonization level and shoot mass, depending on plant family. For the majority of seed families no relationship was observed between colonization level and root mass; however, two seed families showed negative relationships. Shoot height differed only by seed family. Results from our study indicate that the relationship between colonization level and host growth depends on host genotype. We suggest that models of plant intraspecific interactions should consider ectomycorrhizal associations when assessing phenotypic variability.

Effects of NaCl on responses of ectomycorrhizal black spruce (Picea mariana), white spruce (Picea glauca) and jack pine (Pinus banksiana) to fluoride.

Black spruce (Picea mariana), white spruce (Picea glauca) and jack pine (Pinus banksiana) were inoculated with Suillus tomentosus and subjected to potassium fluoride (1 mM KF and 5 mM KF) in the presence and absence of 60 mM NaCl. The NaCl and KF treatments reduced total dry weights in jack pine and black spruce seedlings, but they did not affect total dry weights in white spruce seedlings. The addition of 60 mM NaCl to KF treatment solutions alleviated fluoride-induced needle injury in ectomycorrhizal (ECM) black spruce and white spruce, but had little effect in jack pine seedlings. Both KF and 60 mM NaCl treatments reduced E values compared with non-treated control seedlings. However, with the exception of small reductions of K(r) by NaCl treatments in black spruce, the applied KF and NaCl treatments had little effect on K(r) in ECM plants. Chloride tissue concentrations in NaCl-treated plants were not affected by the presence of KF in treatment solutions. However, shoot F concentrations in ECM black spruce and white spruce treated with 5 mM KF + 60 mM NaCl were significantly reduced compared with the 5 mM KF treatment. The results point to a possible competitive inhibition of F transport by Cl. We also suggest that the possibility that aquaporins may be involved in the transmembrane transport of F should be further investigated.