Effect of mycorrhiza and nitrate nutrition on nitrate reductase activity in Scots pine seedlings

In vivo nitrate reductase (EC 1.6.6.1) activity was measured in seedlings of Scots pine ( Pinus sylvestris L.) inoculated with Cenococcum geophilum (Sow.) Ferd. & Winge, Paxillus involutus (Batsch:Fr) Fr, Piloderma croceum Erikss, & Hjortst, and Suillus variegatus (Fr.) O. Kuntze. The activity was higher in the mycorrhizal pine roots than was previously found in the fungus symbiont alone, but lower than in the roots of nonmycorrhizal pine seedlings. The differences observed in a previous study between the fungal species under pure culture conditions were not found in the present work for mycorrhiza synthezised with the same fungal species. An increase in the nitrate concentration of the nutrient solution increased the proportion of the nitrate reductase activity in the needles. The mycorrhizal root tips had higher nitrate reductase activity than nonmycorrhizal root tips in the same root system.     

Distribution of lead in mycorrhizal and non-mycorrhizal Norway spruce seedlings

Non-mycorrhizal spruce seedlings (Picea abies Karst.) and spruce seedlings colonized with Lactarius rufus (Scop.) Fr. or two strains of Paxillits involutus (Batsch) Fr. were grown in an axenic silica sand culture system with frequently renewed nutrient solution. After successful mycorrhizal colonization, the seedlings were exposed to 1 μM PbCI₂ for 19 weeks. The degree of infection in all of the mycorrhizal treatments approached 100% during the experiment and was not affected by exposure to Pb. However, the number of root tips per root dry weight and the shoot: root ratio, both in the non-mycorrhizal and the mycorrhizal seedlings, had decreased after the 19 week treatment with PbCl₂ Using X-ray microanalysis, the distribution and concentration of Pb in the tissues of mycorrhizal and non-mycorrhizal root tips were compared. In the mycorrhizae of seedlings exposed to Pb no significant accumulation of Pb in the hyphal mantle or in fungal cell walls of the Hartig net were detected. Lead accumulated primarily in the cortex cell walls both of non-mycorrhizal and mycorrhizal root tips. No significant difference of Pb concentrations in root cortex cell walls of non-mycorrhizal and mycorrhizal seedlings was found; except for seedlings colonized with Paxillus involutus strain 537. However, at the endodermis no effect of mycorrhizal fungal colonization on the Pb tissue concentration was detected. The presence of the fungal sheath did not prevent Pb from reaching the root cortex. The endodermis acted as a barrier to Pb radial transport in both mycorrhizal and non-mycorrhizal seedling roots.     

A gnotobiotic culture system with oak microcuttings to study specific effects of mycobionts on plant morphology before, and in the early phase of, ectomycorrhiza formation by Paxillus involutus and Piloderma croceum

Homogeneously developed oak ( Quercus robur L.) microcuttings were challenged in a Petri-dish system with the mycobionts Piloderma croceum J. Erikss. & Hjortst. and Paxillus involutus (Batsch) Fr. Non-destructive observations over 10 wk followed by d. wt measurements at the end of the assays served to precisely characterize root and shoot development, dynamics of mycorrhizal colonization and morphological ratio. In the system, plant development, and especially root morphogenesis, had more similarities to those of stump cuttings or of older seedlings than to those of 3-month-old seedlings. Whereas Paxillus involutus displayed early mycorrhizal colonization and had no significant morphological effects on the host Piloderma croceum modified markedly the entire plant development before a delayed mycorrhiza formation. The latter mycobiont stimulated elongation and production of the lateral root system and also increased the leaf surface. However, no corresponding weight increases were noted, which was reflected by significant increase of both specific root length and specific leaf area. These differential effects are discussed in relation to data concerning carbon requirement and auxin production of the mycobionts. The developed system was shown to be highly suitable for comparative studies with diverse mycobionts on recognition and physiological balance between partners before, and in the early stage of, formation of mycorrhizas.     

Ammonium and methylamine transport by the ectomycorrhizal fungus Paxillus involutus and ectomycorrhizas

Using [(14)C]methylamine as an analogue of ammonium, the kinetics and the energetics of NH(4)(+) transport were studied in the ectomycorrhizal fungus, Paxillus involutus (Batsch) Fr. The apparent half-saturation constant (K(m)) and the maximum uptake rate (V(max)) for the carrier-mediated transport derived from the Eadie-Hofstee transformation were 180 μM and 380 nmol (mg dry wt)(-1) min(-1,) respectively. Both pH dependence and inhibition by protonophores indicate that methylamine transport in P. involutus was dependent on the electrochemical H(+) gradient. Both long-term and short-term uptake experiments were consistent with regulation of ammonium/methylamine transport processes by the presence of an organic nitrogen source. Analysis of methylamine uptake by different P. involutus isolates revealed no obvious trend in the uptake capacities in relation to N deposition at the collection site. Kinetic parameters were determined in P. involutus/Betula pendula (Roth.) axenic association and in detached mycorrhizal roots isolated from forest sites. Enhanced methylamine uptake in the presence of the fungal symbiont was demonstrated. Homogeneous V(max) values were found for axenic and detached mycorrhizas, whereas K(m) values showed greater variations.     

Subcellular compartmentation of elements in non-mycorrhizal and mycorrhizal roots of Pinus sylvestris: an X-ray microanalytical study. II. The distribution of calcium, potassium and sodium

The inter- and intracellular distribution of the elements calcium, potassium and sodium in non-mycorrhizal and mycorrhizal roots of Pinus sylvestris dependent on different external nutrient supply conditions was detected by energy-dispersive X-ray microanalysis after cryofixation, freeze-drying and pressure infiltration of the material. In non-mycorrhizal and mycorrhizal roots, calcium was mainly detectable in the apoplastic regions. The levels in vacuoles and cytoplasm were near the limits of detection by X-ray microanalysis. Incubation with high concentrations of potassium and sodium, or mycorrhizal infection with Suillus bovinus and Pisolithus tinctorius reduced the amounts of calcium detectable in the roots, especially in the apoplast of cortical cells. The studies revealed that: potassium is mainly localized in cytoplasm and cell walls; the cytoplasmic content is regulated over a wide range of external potassium concentrations; potassium levels in the inner parts of roots are higher than in the outer parts. Mycorrhizal infection with Suillus bovinus had no effect on the inter- and intracellular distribution of potassium in roots but, if the external supply was low, the potassium content in shoots was reduced. In non-mycorrhizal pine roots and those infected with Paxillus involutus an increase in the sodium content of all cell compartments was observed after treatment with high external concentrations of NaH₂PO₄. However, an increase in sodium content in mycorrhizas of S. bovinus was not detected. The X-ray microanalytical results are discussed in relation to the apoplastic movement of nutrients in non-mycorrhizal and mycorrhizal fine roots of pine and to the demand for these nutrients in different intracellular compartments.     

A method for establishing ectomycorrhiza on conifer seedlings in steady-state conditions of nutrition

Scots pine ( Pinus sylvestris L.), and Norway spruce ( Picea abies (L.) Karst.) seedlings, inoculated with Suillus bovinits (L. ex Fr.) O. Kuntze and Paxillus involutus (Batsch) Fr., were grown on sloping plastic plates in growth units. A circulating nutrient solution flowed continuously over the plates. Nutrients in balanced proportions previously found to be appropriate for conifers, were added at a specified relative addition rate, in exponentially increasing amounts. The conductivity of the solution was kept low and stable (<50 μS cm⁻¹). No carbohydrates were added.
Seedling nitrogen concentration and relative growth rate attained steady states in both inoculated and uninoculated treatments. The fungi infected the short roots within a few days and formed typical mantles and Hartig nets. This occurred in all treatments, including conditions of free access to nutrients, in which the nutrient concentration of the seedlings was optimal. The growth rate of the extramatrical mycelium was very high.     

Does far-red light affect growth and mycorrhizas of Scots pine seedlings grown in forest soil?

We studied the response of Scots pine (Pinus sylvestris L.) to supplementary far-red sidelight in seedlings grown in a forest soil substrate without additional nutrient supply. Our aim was to determine possible changes in the accumulation and allocation of dry weight and mineral nutrients and the presence of mycorrhizas. Half of the seedlings were grown in light conditions simulating reflected far-red light (FR) from neighbouring plants and the other half were controls not receiving additional FR. PAR irradiance was kept constant in both treatments. At the first harvest (41 d of treatment), FR+ had no effect on stem height, biomass accumulation or allocation. However, at the end of the experiment (93 d of treatment), an increase in stem extension rate and stem dry weight was observed in FR+ seedlings when compared to controls. Both control and treated plants had several morphological types of ectomycorrhizas, but no effect of FR+ on the frequency of these morphotypes was observed. Nor was the concentration of ergosterol or estimated mycorrhizal fungal biomass affected. Nutrients were more responsive to the light quality treatment: P concentration in roots and N and P contents in stems and roots were higher in FR+ plants than in control seedlings. These results are in contrast to those of a previous study in which the root system of pine seedlings, which were fertilised and had less developed mycorrhizas, was reduced by FR+. This revised version was published online in June 2006 with corrections to the Cover Date.     

Mycelial production, spread and root colonisation by the ectomycorrhizal fungi Hebeloma crustuliniforme and Paxillus involutus under elevated atmospheric CO2

Effects of elevated atmospheric carbon dioxide (CO₂) levels on the production and spread of ectomycorrhizal fungal mycelium from colonised Scots pine roots were investigated. Pinus sylvestris (L.) Karst. seedlings inoculated with either Hebeloma crustuliniforme (Bull:Fr.) Quél. or Paxillus involutus (Fr.) Fr. were grown at either ambient (350 ppm) or elevated (700 ppm) levels of CO₂. Mycelial production was measured after 6 weeks in pots, and mycelial spread from inoculated seedlings was studied after 4 months growth in perlite in shallow boxes containing uncolonised bait seedlings. Plant and fungal biomass were analysed, as well as carbon and nitrogen content of seedling shoots. Mycelial biomass production by H. crustuliniforme was significantly greater under elevated CO₂ (up to a 3-fold increase was observed). Significantly lower concentrations and total amounts of N were found in plants exposed to elevated CO₂.     

Apatite as a P source in mycorrhizal and non-mycorrhizal Pinus sylvestris seedlings

The objectives of the study are firstly to test the ability of ectomycorrhizal pine seedlings to use apatite as a P source in comparison with non-mycorrhizal pine seedlings and secondly, to determine if there is a relation between exudation of organic acids and the ability to use apatite as a P source. Non-mycorrhizal Pinus sylvestris (L.) seedlings and seedlings ectomycorrhizal with 4 different isolates of ectomycorrhizal fungi were grown for 220 days in sand/peat filled pots with apatite (Ca₅(F,OH)(PO₄)₃) as the sole P source. In an additional experiment, non-mycorrhizal Pinus sylvestris (L.) seedlings and seedlings ectomycorrhizal with 2 different isolates of ectomycorrhizal fungi were grown without any P source for 250 days. All other nutrients were supplied in a balanced nutrient solution.Ectomycorrhizal seedlings grew less than non-mycorrhizal seedlings but ectomycorrhizal seedlings produced a large external mycelium not included in the biomass estimates. All seedlings in the present study had low shoot:root ratios compared to seedlings growing under optimal conditions. All seedlings grown with apatite as P source had higher foliar P concentrations (0.71–2.11 mg/g) than seedlings growing without any P source (0.57–0.75 mg/g) indicating a significant ability to use apatite as a P source. Seedlings colonized by Suillus variegatus and Paxillus involutus had higher concentrations and total contents of P in shoots compared with non-mycorrhizal seedlings, indicating significant improvement of P uptake by these fungi in comparison with non-mycorrhizal seedlings or seedlings colonized Piloderma croceum.No clear relationship between exudation of organic acids and uptake of P was found. Seedlings colonized by S. variegatus reduced the pH of the soil more than seedlings colonized by P. involutus or non-mycorrhizal seedlings. It is suggested that S. variegatus colonization improves the P uptake by reducing the pH of the soil while P. involutus improves P uptake by having a greater ability to absorb dissolved phosphate than non-mycorrhizal roots or roots colonized by the other fungi used in the study.     

Condition of Scots pine fine roots and mycorrhiza after fungicide application and low-level ozone exposure in a 2-year field experiment

 In a 2-year experiment at an open-air ozone fumigation field, the effects of fungicide application and low-level ozone exposure, single and combined, on fine root and mycorrhiza condition of Scots pine (Pinus sylvestris) seedlings were studied. Two different fungicides, copper oxychloride and propiconazole, with different modes of actions, were used. Propiconazole treatment reduced mycorrhizal infection in both years while copper oxychloride treatment and ozone exposure slightly stimulated mycorrhizal infection after the first year. Different mycorrhizal morphotypes showed different kinds of responses to the two fungicides. Light brown morphotype appeared to be the most sensitive one to propiconazole treatment. After the second year, ectendomycorrhizas disappeared in propiconazole treatment while in control treatment ectendomycorrhizas formed the majority of the light brown morphotype. The root biomass was not affected by fungicide treatments, but ozone exposure increased the total amount of short roots and the fresh weight of propiconazole treated roots. No significant differences in the concentrations of ergosterol, starch and total phenolics in pine roots between treatments were found. However, ergosterol concentration correlated positively with the mycorrhizal infection level. Both fungicides reduced the soil respiration compared to controls. At the ultrastructural level, both fungicides caused increased transparency and gradual granulation and degeneration of cytoplasm in the fungal symbiont of mycorrhizal short roots. Slightly elevated ozone did not have harmful effects on root ultrastructure. These results suggest that fungicides have deleterious effects on the quantity and quality of mycorrhizas in Scots pine roots and also side-effects on non-target soil fungi. Some of these deleterious effects were noticeable only at the ultrastructural level. Received: 23 June 1997 / Accepted: 11 December 1997     

Dynamics of cytoskeletal proteins in developing pine ectomycorrhiza

 Mycorrhizal short roots of Pinus contorta Dougl. ex Loud colonized by Suillus variegatus (Sow. ex Fr.) O. Kuntze or Paxillus involutus (Batsch) Fr. were collected 1–>60 days after fungal contact. The proteins of the inoculated roots were extracted, electrophoretically separated, blotted and immunostained for α-tubulin and actin. The development of the mycorrhiza was also followed microscopically. The signal of plant α-tubulin was stronger than the signal of fungal α-tubulin during the first 5 days in S. variegatus mycorrhiza and was then exceeded by fungal α-tubulin. This correlated well with the increase of fungal mycelium in the mycorrhiza. A transient drop in both plant and fungal α-tubulin signals was observed 20 days after fungal contact, suggesting a change in the metabolism of the mycorrhiza. The signals for plant and fungal actins in the mycorrhiza increased steadily during early infection and then remained at a high level as the mycorrhiza matured. Similar trends were observed in P. contorta–P. involutus mycorrhiza. The data from P. contorta–S.variegatus mycorrhizas suggests that α-tubulin is a growth-related protein, subject to changes, while the amount of actin reflects the general metabolic activity of the mycorrhiza. In both mycorrhizal systems clear α-tubulin and actin signals were detected 60 days after colonization, which indicates that the mycorrhizas were metabolically active in spite of their withered appearance. Accepted: 6 May 1996     

Ectomycorrhizal mycelia reduce bacterial activity in a sandy soil

Abstract: Bacterial activity was studied in a growth system containing Pinus contorta seedlings inoculated with different mycorrhizal fungi. Nylon nets enabled separation of soil compartments with extramatrical mycorrhizal hyphae from soil compartments with roots and mycelium. In three separate experiments bacterial activity, estimated as thymidine incorporation, was reduced in soils with Paxillus involutus hyphae compared to controls without mycorrhizal hyphae. This effect was found irrespective of compartments with and without roots were compared. Laccaria bicolor only reduced the activity in one of these three experiments. Thelephora terrestris (tested in two experiments), Laccaria proxima, Suillus variegatus and Hebeloma crustuliniforme (one experiment), also reduced the thymidine and leucine incorporation rates of bacteria. The reduction for these fungi varied between 20% and 50% in all experiments. Numbers of viable bacteria appeared to be reduced by T. terrestris, L. proxima, S. variegatus and H. crustuliniforme in one experiment, while no effect was seen in the other experiments.     

Rooting and acclimatization of micropropagated cuttings of Pinus pinaster and Pinus sylvestris are enhanced by the ectomycorrhizal fungus Hebeloma cylindrosporum

The effect of different strains of the ectomycorrhizal fungus Hebeloma cylindrosporum on rooting in vitro and acclimatization of micropropagated cuttings of Pinus pinaster and Pinus sylvestris was studied. Two clones of P. pinaster and one of P. sylvestris were unable to root in the absence of auxin, but were induced to root on a medium devoid of auxin by all the fungal strains. Wild-type and indoleacetic acid (IAA)-overproducing mutant strains of the fungus stimulated rooting of clones showing a good reactivity to auxin to the same extent. In contrast, with a clone of P. sylvestris that showed low reactivity to auxin, IAA-overproduction by the fungus was advantageous for the induction of rooting of cuttings. Adventitious roots formed in the presence of a fungal strain were completely surrounded by a loosely packed network of hyphae which formed mycorrhizas as soon as roots grew outside the agar medium. During acclimatization, fungal inoculation improved the survival of rooted cuttings. At the end of acclimatization, fungal mycelia could be easily detected in the culture substrate of cuttings inoculated with dikaryotic strains and most of the pines' short roots were mycorrhizal. Monokaryotic mycelia, which have a lower growth rate and a lower infectivity, displayed poor ability to colonize the substrate and to form mycorrhizas. Two months after the end of acclimatization, fungal inoculation frequently depressed the growth of acclimatized cuttings of the clone J of P. pinaster . No depressive effect was observed with clone 78 and growth stimulation could even be observed with the infective dikaryon D1 which formed numerous mycorrhizas. From these studies, it was concluded that ectomycorrhizal fungi could be a suitable tool for improving rooting in vitro and survival at acclimatization of micropropagated conifer cuttings.     

Media formulation influences in vitro ectomycorrhizal synthesis on the European aspen Populus tremula L.

The effect of various media formulations on in vitro ectomycorrhizal synthesis of identified fungal strains with European aspen (Populus tremula L.) was tested in Petri dishes. Pre-grown seedlings were transferred to various nutrient media and inoculated with Paxillus involutus isolates using modified sandwich techniques. Mycorrhiza formation was evaluated macroscopically and further confirmed by microscopic examination of semi-thin sections for anatomical features of the mantle and the Hartig net. Standard media formulations did not support successful ectomycorrhiza formation because of either very poor plant survival (below 20%) or impaired fungal growth. The inclusion of micronutrients and vitamins in a Melin Norkrans (MMN)-based medium increased plant survival rate to above 60% and supported successful mycorrhizal synthesis. P. involutus isolates formed mycorrhizas with a characteristic Hartig net restricted to the epidermis. Mantle density and thickness varied depending on the isolate. In a follow-up experiment, the adapted medium supported successful ectomycorrhiza formation by various Laccaria and Hebeloma isolates. Our results show that an exogenous supply of vitamins and micronutrients in the medium was a prerequisite for successful mycorrhization of P. tremula in vitro in Petri dishes.     

The extraction and quantification of ergosterol from ectomycorrhizal fungi and roots

Recently, ergosterol analysis has been used to quantify viable fungal biomass in resynthesized ectomycorrhizae. An objective of our study was to quantify ergosterol in a range of ectomycorrhizal isolates under differing growth conditions. In addition, we tested the applicability of the method on field-collected roots of ectomycorrhizal and vesicular-arbuscular (VA) mycorrhizal plants. Quantification of sitosterol as a biomass indicator of plant roots was also undertaken. Ergosterol was not detected in roots of uninoculated Betula populifolia seedlings, and sitosterol was not detected in an ectomycorrhizal fungal isolate but was present in birch roots. Ergosterol was produced in all isolates examined, which represented the major orders of ectomycorrhizal fungi. The range of values obtained, from 3 to nearly 18 g ergosterol mg^-1 dry mass, agrees well with reported values for other mycorrhizal and decomposer fungi. Hyphal ergosterol was the same during growth on phytic acid and KH₂PO₄. Reduction of growth temperature from 25° C to 15° C had little effect on ergosterol content of cultures harvested at similar growth stages. Ergosterol and sitosterol were detected in field-collected ectomycorrhizae of B. populifolia and Pinus sylvestris and VA mycorrhizae of Acer rubrum and Plantago major. Both ergosterol content and ergosterol to sitosterol ratios were significantly lower in VA mycorrhizae than ectomycorrhizae. Calculations of viable fungal biomass associated with field-collected roots were in agreement with those reported by others using the method on resynthesized ectomycorrhizae. Estimates of total mass could be obtained for field-collected B. populifolia roots by a simultaneously using ergosterol to estimate fungal biomass and sitosterol to estimate root mass. Some potential applications and limitations of sterol quantification in studies of mycorrhizal physiology and ecology are discussed.     

The effects of heavy metals,content of nutrients and inoculation with mycorrhizal fungi on the level of terpenoids in roots of <Emphasis Type="Italic">Pinus sylvestris</Emphasis> seedlings

The effect of two substrates differing in the amount of toxic metals and nutrients on the content of volatile organic compounds in roots of Pinus sylvestris seedlings inoculated with Paxillus involutus, Laccaria laccata, Suillus luteus and a strain of the ectendomycorrhizal fungus Mrg X (Ascomycotina) in pot experiment was investigated. Volatile compounds extracted with a supercritical fluid extraction were primarily terpenes and sesquiterpenes, and they were qualitatively the same in roots of mycorrhizal and nonmycorrhizal plants grown on both substrates. The major monoterpenes were α-pinene, Δ³-carene and β-pinene. On the unpolluted substrate, inoculation of plants with the fungi resulted in a similar amount of the volatiles, while on the polluted substrate the mycorrhizal roots showed significantly lower content of terpenoids than nonmycorrhizal roots. The mycorrhizal fungi showed different effects on the concentrations of several terpenoids on both substrates.     

Induced defence responses limit Hartig net formation in ectomycorrhizal birch roots

Roots of clonal birches ( Betula pendula ) were inoculated with the ectomycorrhizal fungi Paxillus involutus (isolates P0 and Mi) and Hebeloma cylindrosporum (strains D1 and D105). These fungi showed different rates of mycorrhiza formation in vitro . Mature mycorrhizas were obtained after only 2–4 d with H. cylindrosporum , whereas 6–8 d were necessary with P. involutus isolate P0, and P. involutus isolate Mi was not able to form mature mycorrhiza during the 10 d of the experiment. Temporal changes in PAL activity and the expression of genes encoding intracellular pathogenesis-related proteins were followed after inoculating birch roots with these fungi. Transient increase of PAL activity, and transient induction of expression of the wound-inducible Bet v1-SC1 gene, were observed in roots challenged with both H. cylindrosporum strains and the P. involutus isolate P0. These changes were found to coincide with hyphal penetration between root cells during Hartig net formation, and were never observed in roots inoculated with the poorly aggressive P. involutus isolate Mi. Examination of mycorrhizal root sections under u.v. light indicated the presence of phenolic compounds in the host cell walls at the vicinity of the Hartig net. These results strongly suggest that hyphal penetration between the root cells triggers a transient defence response which, in turn, could limit Hartig net formation to the outer layer of the root cortex.     

Microcosm-based analyses of Scots pine seedling growth, ectomycorrhizal fungal community structure and bacterial carbon utilization profiles in boreal forest humus and underlying illuvial mineral horizons

We report on the identity of indigenous mycorrhiza forming fungi and rhizosphere/mycorrhizosphere bacterial community carbon source utilization profiles of Scots pine (Pinus sylvestris L.) seedlings grown in boreal forest humus (O) or illuvial (B) mineral horizon containing microcosm growth systems. Based on rDNA (ITS)-RFLP analyses, a total of 10 fungal RFLP taxa were identified from pre-morphotyped mycorrhizas on 7-month-old seedling roots. Hierarchical cluster analysis, including corresponding RFLPs of known fungal species, confirmed root colonization by eight mycorrhizal species. In the O horizon, roots were colonized by e.g. Suillus bovinus, Suillus variegatus, Cenococcum geophilum, Piloderma croceum, Thelephora terrestris and Russula vinicolor. Mycobiont diversity in the mineral B horizon was lower but included Piceirhiza bicolorata and both Suillus species which produced extensive extramatrical mycelium. In comparison to non-colonized soils, rhizosphere and mycorrhizosphere compartments supported significantly higher numbers of bacteria (mean range 10(8)-10(11) cells g(-1) fresh weight (fw)). Specific rhizosphere/mycorrhizosphere 'niche'-linked bacterial communities were detected following multivariate analyses (PCA and CA) of bacterial carbon utilization profiles (Biolog(R) GN microplate). Distinct preferences for amino and carboxylic acids were identified in mineral B horizon rhizospheres whereas a wider range of carbon sources were utilized in the fungal-dominated mycorrhizospheres irrespective of soil types.     

Paxillus involutus Strains MAJ and NAU mediate K(+)/Na(+) homeostasis in ectomycorrhizal Populus x canescens under sodium chloride stress

Salt-induced fluxes of H(+), Na(+), K(+), and Ca(2+) were investigated in ectomycorrhizal (EM) associations formed by Paxillus involutus (strains MAJ and NAU) with the salt-sensitive poplar hybrid Populus × canescens. A scanning ion-selective electrode technique was used to measure flux profiles in non-EM roots and axenically grown EM cultures of the two P. involutus isolates to identify whether the major alterations detected in EM roots were promoted by the fungal partner. EM plants exhibited a more pronounced ability to maintain K(+)/Na(+) homeostasis under salt stress. The influx of Na(+) was reduced after short-term (50 mm NaCl, 24 h) and long-term (50 mm NaCl, 7 d) exposure to salt stress in mycorrhizal roots, especially in NAU associations. Flux data for P. involutus and susceptibility to Na(+)-transport inhibitors indicated that fungal colonization contributed to active Na(+) extrusion and H(+) uptake in the salinized roots of P. × canescens. Moreover, EM plants retained the ability to reduce the salt-induced K(+) efflux, especially under long-term salinity. Our study suggests that P. involutus assists in maintaining K(+) homeostasis by delivering this nutrient to host plants and slowing the loss of K(+) under salt stress. EM P. × canescens plants exhibited an enhanced Ca(2+) uptake ability, whereas short-term and long-term treatments caused a marked Ca(2+) efflux from mycorrhizal roots, especially from NAU-colonized roots. We suggest that the release of additional Ca(2+) mediated K(+)/Na(+) homeostasis in EM plants under salt stress.