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.     

In vitro effects of Laccaria bicolor S238 N and Pseudomonas fluorescens strain BBc6 on rooting of de-rooted shoot hypocotyls of Norway spruce

The ectomycorrhizal fungus Laccaria bicolor S238 N and the bacterium Pseudomonas fluorescens BBc6 were used separately and in combination to induce in vitro rooting of de-rooted shoot hypocotyls of Norway spruce (Picea abies (L.) Karst.). When the culture medium was supplemented with tryptophan, a precursor of indole-3-acetic acid (IAA) synthesis, the presence of the ectomycorrhizal fungus increased the percentage of hypocotyls forming roots; furthermore, both the fungal and bacterial inoculations enhanced the number of roots formed per rooted hypocotyl. Similar results were obtained by adding exogenous IAA (5 and 10 &mgr;M) to the rooting medium. After the rooting phase, the fungal inoculation enhanced adventitious root elongation and branching as well as the aerial growth of the cuttings. Pseudomonas fluorescens BBc6 had no effect on these parameters. The production of IAA by pure cultures of L. bicolor S238 N and P. fluorescens BBc6 was estimated by immunochemical analysis using specific anti-IAA antibodies. Both L. bicolor S238 N and P. fluorescens BBc6 synthesized IAA in pure culture and synthesis was stimulated in the presence of tryptophan. Thus, the effect of the fungus in stimulating adventitious root formation and subsequent elongation and branching can be attributed, at least partially, to the synthesis of IAA by the fungus. The finding that P. fluorescens BBc6 had no effect on root elongation and branching although it produced IAA suggests that either IAA was not the only parameter involved in the stimulation of these processes by L. bicolor S238 N or the bacterium produced other compounds that counteracted the stimulatory effects of IAA on root elongation and branching.     

Effect of ammonium on glutamine synthetase activity in ectomycorrhizal fungi, and in mycorrhizal and non-mycorrhizal Scots pine seedlings

The influence of ammonium on glutamine synthetase activity (GS, EC 6.3.1.2) was studied in three species of ectomycorrhizal fungi, Paxillus involutus (Batsch:Fr) Fr, Piloderma croceum Erikss. and Hjortst. and Suillus variegatus (Fr) O Kuntze growing in pure culture, as well as in the roots and needles of nursery-grown, non-mycorrhizal and mycorrhizal Scots pine (Pinus sylvestris L.) seedlings inoculated with Paxillus involutus or Piloderma croceum as the symbiont. In response to increasing concentrations of ammonium in the nutrient solution, GS activity (expressed on a dry weight basis) increased slightly in Suillus variegatus but not in the other fungi. Glutamine synthetase activity increased in the roots and decreased in the needles of non-mycorrhizal seedlings as the ammonium concentration in the nutrient solution was increased from 0 to 1 mM, but no response was noted with further increases from 1 to 12 mM. Interspecies differences in GS activity were noted among the fungi growing in pure culture, but no significant interspecies differences were observed among the same fungi in the mycorrhizal state.     

Growth and nutrition of nonmycorrhizal and mycorrhizal pitch pine (Pinus rigida) seedlings under phosphorus limitation

The association of ectomycorrhizal fungi with tree roots enhances the acquisition of phosphorus (P) from the soil. In addition to increasing the uptake of H(2)PO(4) (-) (P(i)), mycorrhizal fungi may increase the spectrum of P sources utilized by tree roots by mediating the dissolution of insoluble metallophosphate salts or the hydrolysis of organic P compounds. To investigate the role of ectomycorrhizal fungi in enhancing P acquisition by tree roots, pitch pine (Pinus rigida Mill.) seedlings were grown in sand culture with or without the ectomycorrhizal symbiont Pisolithus tinctorius Coker and Couch under various conditions of P limitation. Compared with nonmycorrhizal seedlings, seedlings inoculated with P. tinctorius exhibited a greater capacity to function under P limitation as evidenced by superior growth and the maintenance of normal foliar ion composition at low P(i) concentrations. Nonmycorrhizal seedlings subjected to P-limiting conditions exhibited depressed K and P and elevated Na concentrations in foliage. The association of P. tinctorius with pitch pine seedling roots maintained foliar K concentrations and prevented the accumulation of Na under P limitation. Nonmycorrhizal seedlings were unable to obtain P from either solid AlPO(4) or inositol hexaphosphate (IHP), whereas seedlings inoculated with P. tinctorius utilized AlPO(4), but not IHP as a P source. Root surface acid phosphatase (APase) activity was depressed in roots infected with the mycorrhizal symbiont and was negatively correlated with seedling growth on all P sources. Root APase activity was negatively correlated with foliar P concentrations in seedlings grown on P(i), but was not correlated with foliar P concentrations in seedlings cultured with AlPO(4) or IHP.     

Binucleate Rhizoctonia (Ceratorhiza spp.) induce adventitious root formation in hypocotyl cuttings of Pinus sylvestris L.

Adventitious rooting of Scots pine (Pinus sylvestris L.) hypocotyl cuttings was promoted by binucleate Rhizoctonia (BnR) and to a lesser extent by ectomycorrhizal fungi. Four BnR isolates (251, 266, 268 and 269) differentiated root meristems and significantly induced adventitious rooting in young derooted seedlings. Rooting rates were significantly higher in BnR treatments than in either indol-3-butyric acid (IBA) pretreatment (200 µm) or co-cultivation with the ectomycorrhizal fungi Suillus bovinus or Laccaria bicolor. In pre-IBA-treated cuttings, adventitious root numbers were higher in the treatments with isolates 251 and particularly 268, while more similar in other BnR treatments. In the 251, roots emerged from distal positions along the hypocotyl and not from the cut base as in the other three BnR isolate treatments. Loss of turgor and hypocotyl wilt was commonly detected up to the point of root initiation. Lateral roots and dichotomous short roots subsequently appeared at high frequency from primary adventitious roots. Mechanisms involved in root meristem differentiation, e.g. auxin production, wound response and oligosaccharide signals, are discussed with respect to host-fungal signalling mechanisms. These beneficial BnR and ectomycorrhizal fungi could provide promising new tools in the development of efficient clonal propagation methodology for this highly recalcitrant Pinus species.     

Influence of elevated CO2 and mycorrhizae on nitrogen acquisition: contrasting responses in Pinus taeda and Liquidambar styraciflua

An understanding of root system capacity to acquire nitrogen (N) is critical in assessing the long-term growth impact of rising atmospheric CO2 concentration ([CO2]) on trees and forest ecosystems. We examined the effects of mycorrhizal inoculation and elevated [CO2] on root ammonium (NH4+) and nitrate (NO3-) uptake capacity in sweetgum (Liquidambar styraciflua L.) and loblolly pine (Pinus taeda L.). Mycorrhizal treatments included inoculation of seedlings with the arbuscular mycorrhizal (AM) fungus Glomus intraradices Schenck & Smith in sweetgum and the ectomycorrhizal (EM) fungus Laccaria bicolor (Maire) Orton in loblolly pine. These plants were then equally divided between ambient and elevated [CO2] treatments. After 6 months of treatment, root systems of both species exhibited a greater uptake capacity for NH4+ than for NO3-. In both species, mycorrhizal inoculation significantly increased uptake capacity for NO3-, but not for NH4+. In sweetgum, the mycorrhizal effect on NO3- and NH4+ uptake capacity depended on growth [C02]. Similarly, in loblolly pine, the mycorrhizal effect on NO3- uptake capacity depended on growth [CO2], but the effect on NH4+ uptake capacity did not. Mycorrhizal inoculation significantly enhanced root nitrate reductase activity (NRA) in both species, but elevated [CO2] increased root NRA only in sweetgum. Leaf NRA in sweetgum did not change significantly with mycorrhizal inoculation, but increased in response to [CO2]. Leaf NRA in loblolly pine was unaffected by either treatment. The results indicate that the mycorrhizal effect on specific root N uptake in these species depends on both the form of inorganic N and the mycorrhizal type. However, our data show that in addressing N status of plants under high [CO2], reliable prediction is possible only when information about other root system adjustments (e.g., biomass allocation to fine roots) is simultaneously considered.     

Ectomycorrhizal fungi affect the physiological responses of Picea glauca and Pinus banksiana seedlings exposed to an NaCl gradient

We tested the effects of ectomycorrhizal (ECM) inoculation on greenhouse-grown white spruce (Picea glauca (Moench) Voss) and jack pine (Pinus banksiana L.) seedlings to be used for revegetation of salt-affected tailing sands resulting from the exploitation of oil sand in northeastern Alberta, Canada. White spruce and jack pine seedlings were inoculated with three ECM fungi selected for their in vitro tolerance to excess Na+ and Cl-: Hebeloma crustuliniforme (Bull) Quel. UAMH 5247, Laccaria bicolor Maire (Orton) UAMH 8232 and a Suillus tomentosus (Kauff.) Sing., Snell and Dick isolate from a salt-affected site. The physiological responses of the seedlings to a gradient of NaCl concentration (0, 50, 100 and 200 mM) were assessed over four weeks by: (1) Na+ accumulation and allocation; (2) chlorophyll a fluorescence; (3) growth, (4) water content; and (5) organic osmolyte accumulation. Jack pine seedlings were more sensitive than white spruce seedlings to increasing Na+ and Cl- concentrations. Both species showed decreasing biomass accumulation, and increasing concentrations of organic osmotica and Na with increasing NaCl concentration. White spruce seedlings inoculated with the S. tomentosus isolate had the best growth response at all NaCl concentrations tested. Although jack pine seedlings inoculated with the L. bicolor or S. tomentosus isolate exhibited the highest growth in the 50 and 100 mM NaCl treatments, both fungi increased the photochemical stress and dehydration of their hosts in the 200 mM NaCl treatment. At the latter concentration, jack pine seedlings inoculated with H. crustuliniforme showed the greatest tolerance to salt stress. Although the different fungi altered the physiological response of the host in different ways, inoculation with salt-stress-tolerant ECM fungi increased growth and reduced the negative effects of excess NaCl. Use of controlled mycorrhization may increase survival of coniferous seedlings used for revegetation of salt-affected sites.     

Prior contact of <Emphasis Type="Italic">Pinus tabulaeformis</Emphasis> with ectomycorrhizal fungi increases plant growth and survival from damping-off

Pinus tabulaeformis (Chinese pine) is a widely planted conifer species in northern China and is used for soil and water conservation on the Loess Plateau. Due to its strong reliance on ectomycorrhizae and low survival rate from damping-off during seedling cultivation and in nurseries, we explored the early influence of three ectomycorrhizal fungi (Handkea utriformis, Suillus lactifluus, and Suillus tomentosus) on the growth, root morphological characters, root vitality, and survival of P. tabulaeformis seedlings from subsequent damping-off in a pot experiment. In addition, the in vitro suppression of three ectomycorrhizal fungi on the damping-off pathogen (Fusarium solani) was evaluated and observed using a scanning electron microscope. We found that all three ectomycorrhizal fungi could colonize more than 40% of the roots of P. tabulaeformis, promote plant shoot and root growth, increase the proportion of large-diameter roots, improve root vitality, and increase survival rates 3 months after inoculation. All three ectomycorrhizal fungi suppressed the growth of F. solani to a different extent in vitro. Ectomycorrhizal fungal hyphae proliferated and wrapped around the hyphae of F. solani when the hyphae encountered each other. The colonization rate of P. tabulaeformis roots was highly correlated with root vitality and root growth parameters, while the survival rate of P. tabulaeformis seedlings was highly correlated with the colonization rate of P. tabulaeformis roots, root vitality and growth. Our results suggest that early contact between P. tabulaeformis and ectomycorrhizal fungi could improve plant growth and resistance against damping-off.     

Shoot water status and ABA responses of transgenic hybrid larch Larix kaempferi x L. decidua to ectomycorrhizal fungi and osmotic stress

It has been postulated that osmotic effects on plant tissue are mediated by abscisic acid (ABA). Hybrid larch (Larix kaempferi (Lambert) Carr. x L. decidua Mill.) plantlets, transformed with the ABA-inducible wheat Em promoter associated with the Gus reporter gene, were axenically inoculated with two ectomycorrhizal fungi: Cenococcum geophilum Fr., considered tolerant to water stress, and Laccaria bicolor (Marie) Orton, considered less tolerant to drought. The mycorrhizal and non-mycorrhizal transgenic plantlets were subjected to osmotic stress by adding polyethylene glycol (PEG) to the culture medium. In the presence of PEG, L. bicolor and C. geophilum reduced shoot water potential and turgor potential, but increased host osmotic potential. Treatment of plantlets with PEG induced a significant increase in endogenous ABA concentrations. Laccaria bicolor and C. geophilum behaved similarly and significantly decreased the ABA response of plantlets to PEG treatment. Moreover, inoculation with either fungus regulated the ABA response of the plantlets even when the fungus was separated from the host by a cellophane sheet that prevented mycorrhiza formation. Although the wheat Em promoter was inducible in larch plantlets, it was not regulated by endogenous ABA. Induction of the wheat Em promoter in larch plantlets depended on organ type, with maximum induction in the root apex. Induction of the Em promoter was significantly decreased by mycorrhizal inoculation.     

Field survival of containerized red and jack pine seedlings inoculated with mycelial slurries of ectomycorrhizal fungi

Red pine and jack pine seedlings growing in styroblocks were inoculated 8 wk after sowing with mycelium/agar slurries of 3 mycorrhizal fungi (Laccaria bicolor, Scleroderma citrinum, and an unidentified basidiomycete), and one suspected mycorrhizal fungus (Cantharellula umbonata). Seedlings inoculated with L. bicolor developed mycorrhizae earlier and in greater numbers than the other inoculation treatments, with red pine out-performing jack pine in both respects. At 34 wk following sowing, seedlings were outplanted on a cleared xeric site in Baraga Co., in Michigan's Upper Peninsula. Seedlings inoculated with C. umbonata failed to form mycorrhizae and were not outplanted. Inoculation treatments did not affect shoot or root weight at outplanting. Red pine inoculated with L. bicolor averaged 21% and 19% greater survival compared with control seedlings after one and two years in the field, respectively. Other inoculation treatments failed to increase seedling survival for either tree species. Jack pine demonstrated higher overall survival than did red pine for both years in all corresponding inoculation treatments.     

Nutritional and pathogenic fungi associated with the pine engraver beetle trigger comparable defenses in Scots pine

Conifer bark beetles are often associated with fungal complexes whose components have different ecological roles. Some associated species are nutritionally obligate fungi, serving as nourishment to the larvae, whereas others are pathogenic blue-stain fungi known to be involved in the interaction with host defenses. In this study we characterized the local and systemic defense responses of Scots pine (Pinus sylvestris L.) against Ophiostoma brunneo-ciliatum Math. (a blue-stain pathogen) and Hyalorhinocladiella macrospora (Franke-Grosm.) Harr. (a nutritional fungus). These fungi are the principal associates of the pine engraver beetle, Ips acuminatus (Gyll.). Host responses were studied following inoculation with the fungi, singly and as a fungal complex, and by identifying and quantifying terpenoids, phenolic compounds and lignin. Although the length of the necrotic lesions differed between control (wound) and fungal treatments, only two compounds (pinosylvin monomethyl ether and (+)-α-pinene) were significantly affected by the presence of the fungi, indicating that Scots pine has a generic, rather than specific, induced response. The fact that both nutritional and blue-stain fungi triggered comparable induced defense responses suggests that even a non-pathogenic fungus may participate in exhausting host plant defenses, indirectly assisting in the beetle establishment process. Our findings contribute to the further development of current theory on the role of associated fungal complexes in bark beetle ecology.     

Ectomycorrhizas and cadmium toxicity in Norway spruce seedlings

We studied the effects of ectomycorrhizal colonization by Laccaria bicolor (Maire) Orton S238 and Paxillus involutus (Batsch) Fr. 533 on cadmium (Cd) toxicity in Norway spruce seedlings (Picea abies (L.) Karst.). Both mycorrhizal and nonmycorrhizal seedlings were exposed to 0 (control), 0.5 or 5 &mgr;M CdSO(4) for 9 weeks in a sand culture system with frequent addition of nutrient solutions. In pure culture, P. involutus and L. bicolor showed similar Cd tolerance. However, in symbiosis, the Cd treatments decreased colonization by L. bicolor but not by P. involutus. Paxillus involutus ameliorated the negative effects of 0.5 &mgr;M Cd on shoot and root growth and chlorophyll content of old needles, whereas L. bicolor did not. Mycorrhizal colonization did not affect Cd concentrations of old needles and roots of seedlings. Despite differences between the ectomycorrhizal fungi in colonization and ability to alleviate Cd toxicity of seedlings, both species reduced Cd concentrations of young needles to a similar degree compared with nonmycorrhizal seedlings. However, in the 0.5 &mgr;M Cd treatment, the Cd content of needles of seedlings colonized by P. involutus was increased, whereas the Cd content of needles of seedlings colonized by L. bicolor was similar to that of needles of nonmycorrhizal seedings. When the amount of Cd translocated to needles was expressed on a root length basis to account for differences in the size of the root systems, the amount of Cd translocated to the needles was similar in seedlings mycorrhizal with P. involutus and in nonmycorrhizal seedlings. All mycorrhizal seedlings were similarly affected by 5 &mgr;M Cd, indicating that the amelioration efficiency of ectomycorrhizal fungi is dependent on the metal concentration to which the roots are exposed. Concentrations of P, K, Ca, Mg and Mn were decreased by 5 &mgr;M Cd to a similar extent in both nonmycorrhizal and mycorrhizal seedlings. In contrast to L. bicolor, P. involutus increased P uptake and altered patterns of root branching. We conclude that mycorrhizas alleviate Cd-induced reductions in growth of Picea abies seedlings. Although the two mycorrhizal fungi examined differed in their ability to alleviate Cd toxicity, these differences were not related to differences in Cd uptake or translocation to the shoot of the mycorrhizal seedlings. We suggest that amelioration of Cd toxicity by P. involutus may be a result of improved P nutrition or changes in root morphology, or both.     

Response of Compensatory-Fertilized Pinus sylvestris to Infection by Heterobasidion annosum

The effects of compensatory fertilization on the infection and growth rate of Heterobasidion annosum were studied in a 40-yr-old, naturally regenerated Scots pine (Pinus sylvestris L.) stand in south-eastern Finland. The treatments were: (1) unfertilized control, (2) a compound fertilizer containing P, K, Ca, Mg, S, Cu, Zn and B, (3) as treatment 2 but with nitrogen, (4) as treatment 3 but with limestone, (5) a mixture of nitrogen and limestone composed on the basis of needle analysis, and (6) wood ash. Pine roots were inoculated on two occasions four and five growing seasons after fertilization using four different strains of the P type of H. annosum. Two years after inoculation, the fungus was alive in 28.9% of the 135 inoculated roots. Fungus survival was highest in the control trees (50%) and poorest in the treatments 2 and 3 (10.5% and 16.7%, respectively). These two treatments differed significantly from the control. Total mycelial extension during the 2-yr study period was 16.4, 2.1, 4.5, 18.1, 15.2 and 5.3 cm in treatments 1, 2, 3, 4, 5 and 6, respectively. Only treatment 2 differed significantly from the control (p<0.05, df=23). The result suggests that the application of slow-release compound fertilizers without supplementary limestone may increase the resistance of Scots pine to infection by H. annosum. The nitrogen-free compound fertilizer may also retard the spread of annosum root rot in infected pine stands.     

Boron and ectomycorrhizal influences on indole-3-acetic acid levels and indole-3-acetic acid oxidase and peroxidase activities of Pinus echinata Mill. roots

The level of indole-3-acetic acid (IAA) in shortleaf pine (Pinus echinata Mill.) roots was increased by inoculation with the ectomycorrhizal fungus Pisolithus tinctorius (Pers.) Coker and Couch. Indole-3-acetic acid oxidase activity was also increased suggesting that the increased level of IAA was the result of increased synthesis. Boron fertilization reduced IAA levels in roots inoculated with P. tinctorius but not in noninoculated roots.     

Growth and nutrient uptake of ectomycorrhizal Pinus sylvestris seedlings in a natural substrate treated with elevated Al concentrations

Models of the effects of elevated concentrations of aluminum (Al) on growth and nutrient uptake of forest trees frequently ignore the effects of mycorrhizal fungi. In this study, we present novel data indicating that ectomycorrhizal mycelia may prevent leaching of base cations and Al. Mycorrhizal and non-mycorrhizal Pinus sylvestris L. seedlings were grown in sand obtained from the B-horizon of a local forest. In Experiment 1, non-mycorrhizal seedlings and seedlings inoculated with Hebeloma cf. longicaudum (Pers.: Fr.) Kumm. ss. Lange or Laccaria bicolor (Maire) Orton were provided with nutrient solution containing 2.5 mM Al. Aluminum did not affect growth of non-mycorrhizal seedlings or seedlings inoculated with L. bicolor. Seedlings colonized by H. cf. longicaudum had the highest biomass production of all seedlings grown without added Al, but the fungus did not tolerate Al. Shoots of seedlings colonized by L. bicolor had the lowest nitrogen (N) concentrations but the highest phosphorus (P) concentrations of all seedlings. The treatments had small but significant effects on shoot and root Al concentrations. In Experiment 2, inoculation with L. bicolor was factorially combined with the addition of a complete nutrient solution, or a solution lacking the base cations K, Ca and Mg, and solutions containing 0 or 0.74 mM Al. Seedling growth decreased in response to 0.74 mM Al, but the effect was significant only for non-mycorrhizal seedlings. Mycorrhizal seedlings generally had higher P concentrations than non-mycorrhizal seedlings. Aluminum reduced P uptake in non-mycorrhizal plants but had no effect on P uptake in mycorrhizal plants. Mycorrhizal colonization increased the pH of the soil solution by about 0.5 units and addition of Al decreased the pH by the same amount. We conclude that the presence of ectomycorrhizal mycelia decreased leaching of base cations and Al from the soil.     

Anatomical-based defense responses of Scots pine (Pinus sylvestris) stems to two fungal pathogens

We investigated the cellular responses of stem tissues of mature Scots pine (Pinus sylvestris L.) trees to inoculations with two fungal pathogens. The bark beetle vectored fungus, Leptographium wingfieldii Morelet, induced longer lesions in the bark, stronger swelling of polyphenolic parenchyma cells, more polyphenol accumulation and increased ray parenchyma activity compared with the root rot fungus, Heterobasidion annosum (Fr.) Bref., or mechanical wounding. Axial resin ducts in the xylem are a general feature of the preformed defenses of Scots pine, but there was no clear induction of additional traumatic axial resin ducts in response to wounding or fungal infection. The anatomical responses of Scots pine to pathogen infection were localized to the infection site and were attenuated away from bark lesions. The responses observed in Scots pine were compared with published studies on Norway spruce (Picea abies (L.) Karst.) for which anatomically based defense responses have been well characterized.     

Light sources with different spectra affect root and mycorrhiza formation in Scots pine in vitro

We studied the effects of broad-spectrum light quality on the interaction between the ectomycorrhizal fungus Pisolithus tinctorius (Pers.) Coker and Couch and Scots pine (Pinus sylvestris L.) seedlings and hypocotyl cuttings cultured in vitro. The light sources were cool white (CW), warm white (WW) and red-rich daylight (RD) fluorescent lamps. Inoculation with P. tinctorius enhanced adventitious root formation of the cuttings in all light treatments. Rooting of the inoculated cuttings was highest in WW light (89%), followed by CW (73%) and RD light (66%). During 6 weeks of in vitro culture, rooted cuttings formed only a few lateral roots. The fungus grew over lateral roots, but the Hartig net was absent in all light treatments. In non-inoculated cuttings, neither root formation nor subsequent root growth was affected by light quality. In the seedling experiment, inoculation in the WW treatment resulted in a significantly (P < 0.05) greater number of lateral roots than inoculation in the RD treatment. The percentage of lateral roots covered with fungal hyphae was also highest in WW light (62%), followed by CW (50%) and RD (27%) light. A similar pattern was observed in the intensity of Hartig net formation. We conclude that effects of broad-spectrum light quality on the ectomycorrhizal fungus-root interaction are dependent on the developmental stage of the root.