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.     

Effects of soil cadmium on Pinus sylvestris L. seedlings

The effects of Cd on the growth and distribution of Cd and mineral nutrients within plant tissues were investigated for Pinus sylvestris L. seedlings grown in mineral forest soil with increasing levels of Cd addition (0–100 mg kg^–1). Approximately 20% of added Cd was found to be extractable from sandy loam forest soil. Root growth was less affected by Cd than shoot growth, which showed a significant reduction in the 100 mg Cd kg^–1 treatment. Cadmium accumulated in roots up to 325 mg kg^–1. Decreased concentrations of K in needles and Ca in stems with increasing Cd levels suggest a disturbance of mineral nutrition as a result of Cd addition.     

Content of phenolics in mycorrhizal roots of Pinus sylvestris seedlings grown in vitro

The effect of Paxillus involutus, Laccaria laccata, Suillus luteus, S. bovinus, Hebeloma crustuliniforme and a strain of the ectendomycorrhizal fungus Mrg X (Ascomycotina) on the content of phenolic compounds: ortho-diphenols and total soluble phenols in roots of Pinus sylvestris seedlings grown in vitro was investigated. Mycorrhizal fungi varied significantly in the ability to influence the concentration of phenolics in Scots pine roots. Plants inoculated with S. luteus accumulated the lowest amount of the phenolics, whereas the amount of phenolics in pine roots inoculated with L. laccata was the highest.     

Zinc-tolerant Suillus bovinus improves growth of Zn-exposed Pinus sylvestris seedlings

Scots pine (Pinus sylvestris L.) seedlings inoculated or not (NM) by a Zn-sensitive or a Zn-tolerant isolate of the ectomycorrhizal fungus Suillus bovinus (L. Fr.) Roussel were exposed to 0.1 or 150 μM Zn²⁺ for 9 months. We hypothesized that inoculation with a Zn-tolerant S. bovinus isolate should result in added Zn resistance of the host plant. Plant and fungal growth as well as nutrient profiles and photosynthetic pigments in pine needles were quantified. In NM plants and in plants colonized by the Zn-sensitive isolate, plant growth, N, P, Mg and Fe assimilation were strongly inhibited under Zn stress and concurred with significantly reduced chlorophyll concentrations. In contrast, plants colonized by the Zn-tolerant isolate grew much better and remained physiologically healthier when exposed to elevated Zn. These results provide further evidence for the important role metal-adapted mycorrhizal fungi play as an effective biological barrier against metal toxicity in trees.     

The effect of ectomycorrhizal fungi on Zn uptake and distribution in seedlings of Pinus sylvestris L

Seedlings of pine, infected with two different ectomycorrhizal fungi, Suillus bovinus (Fr.) O. Kuntze and an unidentified isolate (BP), were exposed to various external Zn concentrations. An additional strain of S. bovinus, cultured on a Zn-enriched medium, was also used. The effect of mycorrhizal associations on Zn uptake and distribution within the plant was determined by AAS.The results show that nonmycorrhizal seedlings have the capability to control the uptake and translocation of Zn to the shoot until the external Zn concentration reaches a threshold level, where no limitation of uptake is possible. Excess Zn is accumulated in the root system to protect the shoot against toxic tissue concentrations. The effect of an ectomycorrhizal infection on Zn uptake and distribution depends on (1) the fungal species (2) the external concentration and (3) the Zn content of the fungal culture medium. Under conditions of low external Zn supply, especially a mycorrhizal infection with S. bovinus led to an increased Zn uptake in root and needles of Pinus sylvestris. Under high external conditions the mycobionts varied considerably in their capability to reduce the transport of Zn to the shoot. Only by an infection with S. bovinus the plant was able to maintain the shoot tissue concentration on a low level. This effect can be enhanced by pretreatment of S. bovinus with high Zn concentrations.     

Conidia of Trichoderma virens as a phosphorus source for mycorrhizal Pinus sylvestris seedlings

In this study, the mobilization and further translocation of phosphorus from conidia of saprotrophic fungus Trichoderma virens into Pinus sylvestris seedlings by nondestructive measuring of ³²P was assessed. The radioactive phosphorus flux from the conidia to the Scots pine seedlings forming mycorrhiza with Laccaria laccata and Suillus bovinus amounted up to 27.82% and 7.42%, respectively, on the 28th day of the experiment, while at the same time in nonmycorrhizal pine seedlings, the detected radioactivity reached only 0.56%. Our studies revealed that both ectomycorrhizal fungi: L. laccata and S. bovinus, mobilized the phosphorus from radioactive conidia of T. virens. On this basis, we conclude that activities of the mycosymbionts may facilitate absorption and further translocation of phosphorus from organic matter into the host plants.     

Concentrations of terpenes in mycorrhizal roots of Scots pine (Pinus sylvestris L.) seedlings grown in vitro

The effect of Paxillus involutus, Laccaria laccata, Suillus luteus, S. bovinus, Hebeloma crustuliniforme and a strain of the ectendomycorrhizal fungus Mrg X (Ascomycotina) on the content of volatile organic compounds in roots of Pinus sylvestris seedlings grown in vitro was investigated. Volatile compounds extracted with a supercritical fluid extraction were primarily terpenes and sesquiterpenes and qualitatively were the same in roots of mycorrhizal and nonmycorrhizal plants. The major monoterpenes were α-pinene, Δ³-carene and β-pinene. Inoculation of plants with the fungi resulted in statistically non-significant increases in the total amount of the volatiles. The mycorrhizal fungi showed diversified effect on the concentrations of several terpenoids.     

Altered root growth and plant chemistry ofPinus sylvestris seedlings subjected to aluminum in nutrient solution

One-year-old Scots pine (Pinus sylvestris L.) seedlings were grown for 9 weeks in nutrient solutions containing 0, 0.5, 1, 2 and 4 mM aluminum nitrate (Al(NO3)3) at pH 4.2. Nine weeks exposure to Al significantly reduced total plant, shoot and root mass and caused a linear decline in proportional allocation of biomass to roots. Relative growth rate of roots declined to as low as zero. Aluminum treatment decreased calcium and magnesium uptake and increased Al content in roots and needles. After 3 weeks of exposure a 10–60% increase in total phenols in roots and a 20–40% increase in o-diphenols in roots and needles were noted. Roots affected by Al showed degeneration of meristematic cells, fewer cell divisions, deformation in cell walls and higher lignification and suberization. The majority of root apices were structurally similar to dormant roots, and a premature senescence of the entire root system was observed. Net photosynthetic rate after 6 weeks of treatment was negatively correlated with needle Al content and Al/Ca ratio (r < -0.9, P < 0.1). The results suggest that Scots pine may be more susceptible to Al than was expected based on previous experiments.     

Survival of inoculated Laccaria bicolor in competition with native ectomycorrhizal fungi and effects on the growth of outplanted Douglasfir seedlings

The survival, development and mycorrhizal efficiency of a selected strain of Laccaria bicolor along with naturally occurring ectomycorrhizal fungi in a young plantation of Douglas fir was examined. Symbionts were identified and their respective colonization abilities were determined. Eight species of symbiotic fungi, which may have originated in adjacent coniferous forests, were observed on the root systems. Mycorrhizal diversity differed between inoculated (5 taxa) and control (8 taxa) seedlings. Ectomycorrhizal fungi which occurred naturally in the nursery on control seedlings (Thelephora terrestris and Suillus sp.) did not survive after outplanting. Both inoculated and naturally occurring Laccaria species, as well as Cenococcum geophilum, survived on the old roots and colonized the newly formed roots, limiting the colonization by other naturally occurring fungi. Other fungi, such as Paxillus involutus, Scleroderma citrinum and Hebeloma sp. preferentially colonized the old roots near the seedling's collar. Russulaceae were found mainly in the middle section of the root system. Mycorrhizal colonization by Laccaria species on inoculated seedlings (54%) was significantly greater than on controls (13%) which were consequently dominated by the native fungi. Significant differences (up to 239%) were found in the growth of inoculated seedlings, especially in root and shoot weight, which developed mainly during the second year after outplanting. Seedling growth varied with the species of mycorrhizae and with the degree of root colonization. Competitiveness and effectiveness of the introduced strain on improving growth performances of seedlings are discussed.     

Effects of elevated CO2 and nitrogen on nutrient uptake in ponderosa pine seedlings

This paper reports on the results of a controlled-environment study on the effects of CO₂ (370, 525, and 700 mol mol^-1) and N [0, 200, and 400 g N g soil^-1 as (NH₄)SO₄] on ponderosa pine (Pinus ponderosa) seedlings. Based upon a review of the literature, we hypothesized that N limitations would not prevent a growth response to elevated CO₂. The hypothesis was not supported under conditions of extreme N deficiency (no fertilizer added to a very poor soil), but was supported when N limitations were less severe but still suboptimal (lower rate of fertilization). The growth increases in N-fertilized seedlings occurred mainly between 36 and 58 weeks without any additional N uptake. Thus, it appeared that elevated CO₂ allowed more efficient use of internal N reserves in the previously-fertilized seedlings, whereas internal N reserves in the unfertilized seedlings were insufficient to allow this response. Uptake rates of other nutrients were generally proportional to growth. Nitrogen treatment caused reductions in soil exchangeable K⁺, Ca²⁺, and Mg²⁺ (presumably because of nitrification and NO₃ ^- leaching) but increases in extractable P (presumably due to stimulation of phosphatase activity).The results of this and other seedling studies show that elevated CO₂ causes a reduction in tissue N concentration, even under N-rich conditions. The unique response of N is consistent with the hypothesis that the efficiency of Rubisco increases with elevated CO₂. These results collectively have significant implications for the response of mature, N-deficient forests to evevated CO₂.     

Sensitivity to Cd or Zn of host and symbiont of ectomycorrhizal Pinus sylvestris L. (Scots pine) seedlings

Pinus sylvestris seedlings infected with either the ectomycorrhizal (ECM) fungus Paxillus involutus or Suillus variegatus were exposed to a range of Cd or Zn concentrations. This was done to investigate the relationship between the sensitivity of ECM fungi and their host plants over a wide range of concentrations. P. involutus ameliorated the toxicity of Cd and Zn to P. sylvestris with respect to root length, despite significant inhibition of ECM infection levels by Cd (Cd EC₅₀ [effective concentration which inhibits ECM infection by 50%] values were: P. involutus 3.7 μg g^-1 Cd; S. variegatus 2.3 μg g^-1 Cd). ECM infection by P. involutus also decreased Cd and Zn transport to the plant shoots at potentially toxic concentrations and also influenced the proportion of Zn transported to the roots and shoots, with a higher proportion retained in the roots of the seedlings. ECM infection did increase host biomass production, but this was not affected by the presence of Cd or Zn. Root and shoot biomass production by P. sylvestris, in both the presence and absence of ECM fungi, was unaffected by Cd and Zn at all concentrations tested. This revised version was published online in August 2006 with corrections to the Cover Date.     

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.