Differences in 15N uptake amongst spruce seedlings colonized by three pioneer ectomycorrhizal fungi in the field

Amongst the factors hypothesized to be responsible for high ectomycorrhizal fungal diversity are resource partitioning and niche differentiation. However, functional differences amongst ectomycorrhizal fungi, which are pre-requisites for resource partitioning, are known primarily from lab studies; now realistic field experiments are needed in order to establish that these differences exist under field conditions. In this study, Picea engelmannii seedlings planted in a subalpine clearcut became naturally colonized over the course of 1 y. Then a defined volume of soil around each seedling was injected with ¹⁵N-labelled nitrate, ammonium or aspartate. Seedling biomass and N content increased, but N concentration decreased, with percent colonization of root systems. Accumulation of ¹⁵N per unit dry weight was not affected by the proportion of roots colonized but, rather, was influenced by the primary ectomycorrhizal fungus colonizing the seedling. Seedlings colonized by a Wilcoxina sp. accumulated more ¹⁵N per g than seedlings colonized by a Cenococcum sp. The presence of dark septate hyphae in the mantle was associated with lower accumulation of ¹⁵N by seedlings colonized by Amphinema byssoides. Our results demonstrate that the physiological differences required to support the concept of niche differentiation amongst ectomycorrhizal fungi exist in the field.     

Ectomycorrhizal Specificity Patterns in a Mixed Pinus contorta and Picea engelmannii Forest in Yellowstone National Park

We used molecular genetic methods to test two hypotheses, (i) that host plant specificity among ectomycorrhizal fungi would be common in a closed-canopy, mixed Pinus contorta-Picea engelmannii forest in Yellowstone National Park and (ii) that specificity would be more common in the early successional tree species, P. contorta, than in the invader, P. engelmannii. We identified 28 ectomycorrhizal fungal species collected from 27 soil cores. The proportion of P. engelmannii to P. contorta ectomycorrhizae was nearly equal (52 and 48%, respectively). Of the 28 fungal species, 18 composed greater than 95% of the fungal community. No species was associated exclusively with P. contorta, but four species, each found in only one core, and one species found in two cores were associated exclusively with P. engelmannii. These fungi composed less than 5% of the total ectomycorrhizae. Thus, neither hypothesis was supported, and hypothesized benefits of ectomycorrhizal specificity to both trees and fungi probably do not exist in this system.     

Role of mycorrhizal networks and tree proximity in ectomycorrhizal colonization of planted seedlings

The importance of mycorrhizal network (MN)-mediated colonization under field conditions between trees and seedlings was investigated. We also determined the combined influences of inoculum source and distance from trees on the ectomycorrhizal (EM) community of seedlings. On six sites, we established trenched plots around 24 residual Pseudotsuga menziesii var. glauca trees and then planted seedlings at four distances (0.5, 1.0, 2.5, and 5.0 m) from the tree into four mesh treatments that served to restrict inoculum access (i.e., planted into mesh bags with 0.5, 35, 250 μm pores or directly into soil). Ectomycorrhizal communities were identified after two growing seasons using morphological and molecular techniques. Mesh treatments had no effect on seedling mycorrhizal colonization, richness, or diversity, suggesting that MN-mediated colonization, was not an essential mechanism by which EM communities were perpetuated to seedlings. Instead, wind-borne and soil inoculum played an important role in seedling colonization. The potential for MNs to form in these forests was not dismissed, however, because trees and seedlings shared 83 % of the abundant EM. Seedlings furthest from trees had a simpler EM community composition and reduced EM richness and diversity compared to seedlings in closer proximity.     

Response of ectomycorrhizalPinus banksiana andPicea glauca to heavy metals in soil

Pinus banksiana andPicea glauca inoculated or not with the ectomycorrhizal fungusSuillus luteus were grown in a sandy loam soil containing a range of Cd, Cu, Ni, Pb and Zn concentrations. Ectomycorrhizal colonization rates were significantly reduced on Pinus and Picea seedlings by the heavy metals, particularly Cd and Ni. Needle tissue metal concentrations were lower in ectomycorrhizal seedlings at low soil metal concentrations. However, at higher soil concentrations, heavy metal concentrations of needle tissue were similar in ectomycorrhizal and nonmycorrhizal plants. The growth of nonmycorrhizal seedlings exposed to heavy metals was reduced compared to those inoculated withSuillus luteus. Apparently ectomycorrhizal colonization can protect Pinus and Picea seedlings from heavy metal toxicity at low or intermediate soil concentrations of Cd, Cu, Ni, Pb and Zn.     

The co-occurrence of ectomycorrhizal,arbuscular mycorrhizal,and dark septate fungi in seedlings of four members of the Pinaceae

Although roots of species in the Pinaceae are usually colonized by ectomycorrhizal (EM) fungi, there are increasing reports of the presence of arbuscular mycorrhizal (AM) and dark septate endophytic (DSE) fungi in these species. The objective of this study was to determine the colonization patterns in seedlings of three Pinus (pine) species (Pinus banksiana, Pinus strobus, Pinus contorta) and Picea glauca x Picea engelmannii (hybrid spruce) grown in soil collected from a disturbed forest site. Seedlings of all three pine species and hybrid spruce became colonized by EM, AM, and DSE fungi. The dominant EM morphotype belonged to the E-strain category; limited colonization by a Tuber sp. was found on roots of Pinus strobus and an unknown morphotype (cf. Suillus–Rhizopogon group) with thick, cottony white mycelium was present on short roots of all species. The three fungal categories tended to occupy different niches in a single root system. No correlation was found between the percent root colonized by EM and percent colonization by either AM or DSE, although there was a positive correlation between percent root length colonized by AM and DSE. Hyphae and vesicles were the only AM intracellular structures found in roots of all species; arbuscules were not observed in any roots.     

Growth of mycorrhizal jack pine (Pinus banksiana) and white spruce (Picea glauca) seedlings planted in oil sands reclaimed areas

The effectiveness of ectomycorrhizal inoculation at the tree nursery seedling production stage on growth and survival was examined in jack pine (Pinus banksiana) and white spruce (Picea glauca) planted in oil sands reclamation sites. The seedlings were inoculated with Hebeloma crustuliniforme strain # UAMH 5247, Suillus tomentosus strain # UAMH 6252, and Laccaria bicolor strain # UAMH 8232, as individual pure cultures and in combinations. These treatments were demonstrated to improve salinity resistance and water uptake in conifer seedlings. The field responses of seedlings to ectomycorrhizal inoculation varied between plant species, inoculation treatments, and measured parameters. Seedling inoculation resulted in higher ectomycorrhizal colonization rates compared with non-inoculated control, which had also a relatively small proportion of roots colonized by the nursery contaminant fungi identified as Amphinema byssoides and Thelephora americana. Seedling inoculation had overall a greater effect on relative height growth rates, dry biomass, and stem volumes in jack pine compared with white spruce. However, when examined after two growing seasons, inoculated white spruce seedlings showed up to 75 % higher survival rates than non-inoculated controls. The persistence of inoculated fungi in roots of planted seedlings was examined at the end of the second growing season. Although the inoculation with H. crustuliniforme triggered growth responses, the fungus was not found in the roots of seedlings at the end of the second growing season suggesting a possibility that the observed growth-promoting effect of H. crustuliniforme may be transient. The results suggest that the inoculation of conifer seedlings with ectomycorrhizal fungi could potentially be carried out on a large scale in tree nurseries to benefit postplanting performance in oil sands reclamation sites. However, these practices should take into consideration the differences in responses between the different plant species and fungal strains.     

连续三年夜间增温和施氮对云杉外生菌根及菌根真菌多样性的影响

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

Habitat matters: The role of spore bank fungi in early seedling establishment of Florida slash pines

This study investigated broad patterns in communities of ectomycorrhizal fungi from three Florida habitats (sandhills, scrub, and pine rocklands) and the ability of spore bank fungi to associate with Pinus elliottii (slash pine) and Pinus densa (south Florida slash pine). Efforts to replant pines in the endangered pine rocklands are vital to the persistence of this habitat, yet little is known about the ectomycorrhizal fungi communities or how they may differ from those in other pine-dominated habitats in Florida. We used high-throughput amplicon sequencing (HTS) to assess baseline fungal communities and greenhouse bioassays to bait ectomycorrhizal fungi using seedlings. HTS soil data recovered 188 ectomycorrhizal species but only a few subsequently colonized the bioassay seedlings. We recovered 21 ectomycorrhizal species on pine seedlings including common spore bank fungi such as Cenococcum, Suillus, and Tuber, but Rhizopogon species were dominant across all sites and habitats. Habitat type and site were significant variables influencing the community composition of the total soil fungal community, soil ectomycorrhizal community, and the fungi found on seedling root tips. However, we found no significant differences between the ectomycorrhizal communities on seedling roots from the two Pinus species.     

Influence of ectomycorrhizal inoculation on Pinus wallichiana and Cedrus deodara seedlings under nursery conditions

A study was undertaken to examine the extent of root colonization by four locally isolated ectomycorrhizal （ECM） fungi （Hebeloma theobrominum, Boletus dryophilus, Scleroderma citrinum and Suillus luteus） and their effects on seedling growth in Pinus wallichiana and Cedrus deodara under nursery conditions. Seedlings of the two conifers were inoculated with mycelium of ECM fungi and were grown in pots containing sterilized forest soil for six months. The percentage of ECM colonization of roots was 38%-52% in Pinus wallichiana and 33%-48~ in Cedrus deodara. ECM colonization increased shoot height, needle number, shoot and root biomass and survival of inoculated seedlings. Among the four ECM fungi Hebeloma theobrominum was more effective with Pinus wallichiana and Scleroderma citrinum with Cedrus deodara in promoting seedling survival and overall growth. All the four ECM fungi used enhanced growth of inoculated seedlings and thus can be used in afforestation and regeneration programmes in degraded forests ecosystems.     

Positive effects of ectomycorrhizal colonization on growth of seedlings of a tropical tree across a range of forest floor light conditions

In a shadehouse experiment we tested the effects of light, nutrients and ectomycorrhizal fungi (EMF) on the growth of Vatica albiramis van Slooten (Dipterocarpaceae) seedlings. We hypothesised that it is more advantageous for plants to form connections with EMF and to trade carbon for nutrients with EMF under high light than low light. The relationship between seedling growth and the proportion of ectomycorrhizal root tips was expected as positive in high light and as negative in low light. Light conditions simulated the forest understory (low; 3% full sunlight), a small gap (medium; 11%) and a large gap (high; 33%) and a fully factorial combination of nutrients (F?/+) and ectomycorrhizal colonization (EMF?/+) treatments were applied within light conditions. The application of EMF and nutrients did significantly alter seedling growth across the range of forest floor light conditions, however the key hypothesis was rejected as seedling growth under low light was not affected by increased EMF colonization of root tips (light:EMF colonization χ²?=?2.97, p?=?0.23). In addition, the lack of difference in morphotype abundance across light conditions indicated that light changes may not favour the association to specific EMF in seedlings of this particular dipterocarp species. Our results suggest that antagonistic (non-beneficial to the plant) effects due to ectomycorrhizal colonization under a light constrained environment may not affect seedling growth of Vatica albiramis.     

The effect of forest soil and community composition on ectomycorrhizal colonization and seedling growth

Plant–soil feedbacks have been observed in many forest communities, but the role of the mycorrhizal community in perpetuating feedback loops is still poorly understood. Mycorrhizal community composition is closely linked to soil properties and host plant composition, which highlights their potential importance in plant–soil–fungus loops. Eastern hemlock (hemlock; Tsuga canadensis) seedlings were grown in soil bioassays in growth chambers and transplanted under closed forest canopy to examine the effect of hardwood and hemlock forest soil on seedling growth, survival, and ectomycorrhizal fungi (EMF) colonization. Seedlings propagated in hemlock forest soil had greater height growth compared with sterile control soil and achieved greater mycorrhizal colonization than seedlings grown in hardwood forest soils after 9 months in a growth chamber. Outplanted seedlings grown in hemlock communities achieved significantly greater increment growth than those seedlings grown in hardwood communities (mean height difference (95% CI)?=?0.39 cm (0.14–0.63 cm)), although final survival and EMF colonization was similar between forest types. EMF diversity (Shannon-Wiener index (SE)?=?1.88 (0.28) and 1.23 (0.44) for hardwood and hemlock, respectively) and community assemblage (Jaccard index (SE)?=?19.0% (4%)) differed between the two forest communities. EMF community assemblage was associated with both the forest type (i.e. plant community/microsite effects) and initial soil type (i.e. soil characteristics/resistant inoculum). The results support previously observed positive feedbacks between conspecifics under hemlock forest communities and provides evidence for the role of the EMF community within this feedback loop. Alternatively, the reduced growth of hemlocks under hardwoods may be attributed to the different EMF community associated with that forest.     

Structure and activities of ectomycorrhizal and microbial communities in the rhizosphere of Fagus sylvatica under ozone and pathogen stress in a lysimeter study

The aim was to study the influence of abiotic (elevated ozone) or biotic stress (Phytophthora citricola) or their combination on soil biological components and processes in the rhizosphere of young beech trees. Ectomycorrhizal and overall microbial community composition was studied at two soil depths in a lysimeter experiment with 7 year old trees of Fagus sylvatica. As a functional parameter, potential enzyme activities were measured in mycorrhizosphere soil and on excised mycorrhizal tips. The degree of mycorrhization, structure and potential enzymatic activities of mycorrhizal communities were only slightly influenced by treatments. Soil enzyme activities were depressed under elevated ozone and stimulated by P. citricola under ambient but not under elevated ozone. Overall microbial community composition (PLFA) and ectomycorrhizal diversity changed with depth. PLFA analyses not only suggested a reaction of the microbial community to elevated ozone but also indicated an increase in plant stress related components. No influence of the biotic stress on ectomycorrhizal or overall microbial community structure was detected. Changes in the mycorrhizosphere community structure and function due to ozone may be explained by the quality of plant derived carbon.     

The effect of mercury on the establishment of Pinus rigida seedlings and the development of their ectomycorrhizal communities

Metals are toxic to both plants and fungi, and elevated soil metal concentrations have been documented to change the structure of ectomycorrhizal communities. Mercury (Hg) is a highly toxic metal and inhibits the growth of ectomycorrhizal fungi (ECMF) in axenic culture. However, the effects of Hg on the growth of tree seedlings and the development of their ECMF communities have not been explored. In the current study, Pinus rigida seedlings were planted in soil amended with 0–366 μg g^?1 Hg and incubated for 5 months. Survival and growth of P. rigida seedlings was determined, and their ECMF communities were characterized by morphotype analysis. Seedling survival declined with increasing Hg additions, although no reduction in growth was detected among surviving seedlings. The addition of 88 μg g^?1 Hg to soil more than halved the total ectomycorrhizal colonization of root tips and reduced both richness and diversity of root tip morphotypes, while lower Hg additions did not significantly affect ECMF community composition relative to the no Hg control. This suggests that changes in the community of ECMF may occur in contaminated soils before any aboveground effects on surviving seedlings are noticeable, potentially altering the contribution of ECMF to the fitness of established host plants.     

Soil inoculation of lodgepole pine seedlings alters root-associated fungal communities but does not improve seedling performance in beetle-killed pine stands

Extensive tree mortality in forests can change the community composition of soil fungi altering seedling establishment, a process critical to forest restoration. Disturbances that result in the loss of ectomycorrhizal fungi, in particular, may impede the establishment of tree species reliant on these symbionts for their survival. Inoculation of seedlings with soil from intact forests may improve the establishment of seedlings in such disturbances but the method has rarely been tested in the field. Here, we assess whether soil inoculation improves lodgepole pine (Pinus contorta var. latifolia) seedling performance in conspecific stands with high levels of tree mortality caused by a mountain pine beetle (Dendroctonus ponderosae) outbreak and whether underlying soil type modifies inoculation effects. We first inoculated seedlings in a growth chamber with small amounts of soils (5% volume) originating from either intact (<10%) or “beetle-killed” (>70% pine basal area killed) conspecific stands or added no soil inoculum and, after 4 months, transplanted them into 15 beetle-killed stands. After two growing seasons, root-associated fungal communities of seedlings receiving inoculum from intact stands differed in composition from those receiving inoculum from beetle-killed stands or no inoculum. However, inoculation had no effect on seedling survival, height, or biomass. Site properties, including soil texture and the resident fungal community composition, overwhelmed the effect of soil inoculation on seedling performance. Seedling survival and shoot mass was higher in sandy than loamy soils. Restoration to improve seedling performance in beetle-killed stands should consider stand-level treatments as soil inoculation at the level evaluated was ineffective.     

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 sea level rise on mangrove Avicennia population growth,colonization and establishment: Evidence from a field survey and greenhouse manipulation experiment

Mangroves will either face longer tidal inundation or retreat landwards in response to on-going accelerated sea level rise. However, little is known about the growth, regeneration or colonization of mangrove seedlings under the different tidal inundation regimes associated with accelerated sea level rise. In the present study, a field survey and a greenhouse mesocosm experiment were conducted to evaluate possible effects of accelerated sea level rise on colonization, establishment and seedling growth of a mangrove pioneer species, Avicennia marina. Avicennia populations at different elevations of the intertidal zone on Xiamen Island in Fujian Province, China showed distinctive patterns in both seedling density and plant age. The seedlings at lower elevations had less annual biomass accumulation and population productivity, but higher shoot to root ratios, suggesting that elevation has positive effects on seedling growth. The greenhouse mesocosm experiment with 1-year-old A. marina seedlings utilized five inundation periods (0, 2, 4, 6 and 12 h in a semidiurnal tidal cycle) and two inundation depths (root immersion and canopy immersion). Both inundation period and depth exerted significant and negative effects on biomass accumulation, photosynthetic rate, leaf electron transportation and water use efficiency. However, the negative effects of canopy immersion were more profound than root immersion. Canopy immersion exacerbated the effects of prolonged inundation, with no seedlings surviving under canopy immersion at the 12-hour treatment. These results suggest that at lower elevations with higher sea level, canopy immersion will have greater negative effects on seedling colonization, establishment and early growth of A. marina. This finding is instrumental in predicting the future dynamics of mangrove forests under increasing sea levels.     

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

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

Nutrient and Carbon Additions to the Microbial Soil Community and its Impact on Tree Seedlings in a Boreal Spruce Forest

We have added glucose and nutrients to manipulate soil microbial activity and nutrient availability in a boreal spruce forest to study the performance of birch and spruce seedlings in relation to the soil microbial community. The proportion of aboveground biomass in the seedlings was largest in plots amended with extra nutrients, while ectomycorrhizal (ECM) colonisation was low in these plots. ECM appeared beneficial for growth of both species, but only at low levels of colonisation (<25% ECM colonised root-tips). The soil microbial biomass, as determined by total PLFA, was largest in plots treated with glucose and there was a significant negative relationship between birch seedling size and levels of total PLFA in soil. This could be taken to suggest that poor seedling growth was due to nutrient limitation caused by microbial assimilation. However, the treatment response of the birch seedlings was generally weak, and spruce often showed no response at all to the addition of nutrients and glucose. The most consistent parameter for the variation in plant performance, as well as for the microbial soil community, was the block-effect. This suggests a strong spatial structure in the soil microbial community, and that this structure was robust with respect to our treatments even though they continued over a 3-year period.     

Effects of isolates of ectomycorrhizal fungi and endophytic Mycelium radicis atrovirens that were dominant in soil from disturbed sites on growth of Betula platyphylla var. japonica seedlings

Effects of ectomycorrhizal fungi and endophytic Mycelium radicis atrovirens Melin (MRA) on growth of Betula platyphylla var. japonica seedlings were investigated under aseptic culture conditions. Three isolates of ectomycorrhizal fungi and two isolates of MRA were used. One MRA isolate was Phialocephala fortinii. Previous field work revealed that these isolates were dominant on the roots of B. platyphylla var. japonica seedlings grown in a mineral subsoil that had been exposed by the removal of surface soil. After a 100-day incubation, the growth of the seedlings was significantly enhanced by the colonization of these ectomycorrhizal fungal isolates as compared with uninoculated seedlings. In contrast, the growth of seedlings was retarded by the colonization of the MRA isolates. The growth of seedlings that were co-inoculated with ectomycorrhizal fungi and MRA was similar to that of uninoculated seedlings in most cases. These results suggest that ectomycorrhizal fungi have a beneficial effect on the growth of B. platyphylla var. japonica seedlings and that they suppress the deleterious effect of MRA. Thus, these ectomycorrhizal fungi probably have an important role in establishing B. platyphylla var. japonica seedlings during the initial stage of re-vegetation following site disturbance by the removal of surface soil.     

Effects of growth medium, nutrients, water, and aeration on mycorrhization and biomass allocation of greenhouse-grown interior Douglas-fir seedlings

Commercial nursery practices usually fail to promote mycorrhization of interior Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco var. glauca (Beissn.) Franco] seedlings in British Columbia, which may account for their poor performance following planting in the field. We tested the effects of four nursery cultivation factors (nitrogen fertilization, phosphorus fertilization, watering, and soil aeration) and field soil addition on mycorrhization, survival, growth, and biomass allocation of interior Douglas-fir seedlings in a series of greenhouse experiments. Where field soil was added to the growing medium, mycorrhization and root/shoot ratios were maximized at lower levels of mineral nutrient application and aeration. Where field soil was not added, mycorrhization was negligible across all fertilization and aeration treatments, but root/shoot ratio was maximized at lower levels of mineral nutrients and the highest level of aeration. Regardless of whether field soil was added, intermediate levels of soil water resulted in the best mycorrhizal colonization and root/shoot ratios. However, field soil addition reduced seedling mortality at the two lowest water levels. A cluster analysis placed ectomycorrhizal morphotypes into three groups (Mycelium radicis-atrovirens Melin, Wilcoxina, and mixed) based on their treatment response, with all but two morphotypes in the mixed group whose abundance was maximized under conditions common to advanced seedling establishment. For maximal mycorrhization and root development of interior Douglas-fir seedlings, nurseries should minimize addition of nitrogen and phosphorus nutrients, maximize aeration, provide water at moderate rates, and, where possible, add small amounts of field soil to the growing medium.