Effects of soil-applied lead on seedling growth and ectomycorrhizal colonization of loblolly pine

Six-month-old loblolly pine (Pinus taeda) were grown for 15 weeks in two native soils amended with 0, 30, 60, 120, 240 or 480 mg kg(-1) Pb as PbCl2. Ectomycorrhizae were quantified, by morphotype, as the total number of tips per centimeter, and as the number of tips for each morphotype and for Cenococcum geophilum. Total numbers of non-ectomycorrhizal short roots and necrotic tips were recorded. Total height and biomass exhibited a non-linear response to soil-applied lead. Growth generally was greatest in the controls and higher treatments, and least in the intermediate treatments. In both soils, Pb concentrations in roots increased linearly with increasing levels of soil-applied Pb. Neither foliage nor stems exhibited significant increases in Pb concentrations with increasing levels of Pb. Significant linear decreases in total numbers of ectomycorrhizal tips and significant linear increases in non-ectomycorrhizal short roots and necrotic tips occurred with increasing levels of Pb in the soil. The majority of individual morphotypes decreased with increasing Pb. However, the number of ectomycorrhizal tips formed by C. geophilium increased with increasing soil Pb levels after 15 weeks of treatment. These results indicate that short-term loblolly pine seedling growth is not inhibited by increasing Pb levels. Ectomycorrhizal formation decreased, and alterations in species composition occurred as a result of increasing concentrations of soil-applied Pb. These effects on ectomycorrhizae may cause long-term changes in nutrient and water balances, which could reduce tree vigor.     

Effects of simulated rain acidity on ectomycorrhizae of red spruce seedlings potted in natural soil

Formation of ectomycorrhizae of red spruce (Picea rubens) grown in natural soil was measured after seedlings were exposed to 25 or 50 applications of simulated rain of pH 5.5, 3.5 or 2.5. Ectomycorrhizae were quantified as the total number of ectomycorrhizal tips per centimeter, and as the number of ectomycorrhizal tips for each morphotype and for Cenococcum geophilum. Rain solutions were applied to the soil alone, to foliage and stem alone, or to entire potted seedlings. Final soil pH was linearly related to rain solution acidity. Lower base saturation, calcium and zinc content, and higher exchangeable acidity were observed after pH 2.5 treatments if the soil was exposed. Rain solutions and the subsequent changes in soil characteristics did not affect the total numbers of ectomycorrhizal tips. Four morphotypes of ectomycorrhizae observed for these seedlings were unaffected by simulated rain. However, the numbers of ectomycorrhizal tips formed by C. geophilum tended to increase with rain solution acidity after 50 applications. Method of rain deposition did not affect ectomycorrhizae, suggesting both plant and soil mediated responses may favor certain mycobionts. The results of this study indicate that short-term acidic deposition does not induce significant changes in the frequency of ectomycorrhizae, but higher numbers of C. geophilum tips suggest there may be changes in the relative occurrence of specific morphotypes of fungus species.     

Ectomycorrhizal colonization of loblolly pine seedlings during three growing seasons in response to ozone, acidic precipitation, and soil Mg status

A three-year study was initiated in 1987 to evaluate the impact of O3, acidic precipitation, and soil Mg on ectomycorrhizal colonization of loblolly pine (Pinus taeda L.) seedlings. Thirty-six open-top chambers equipped with a rainfall exclusion/addition system were utilized to administer three levels of O3 (subambient, ambient, or twice ambient) and two precipitation acidity levels (pH 3.8 or 5.2) to seedlings growing in 24-liter plastic pots containing soil having either 35 or 15 mg kg(-1) of exchangeable Mg. Seedlings exposed to the twice ambient O3 treatment exhibited smaller percentages of total ectomycorrhizal short roots at the end of each year of the study, but trends were statistically significant in 1989 only. Changes in number of specific ectomycorrhizal morphotypes in response to O3 were not consistent from year to year. Acidic precipitation treatments had no effect on number or percent of mycorrhizal short roots, and responses of two morphotypes to soil Mg treatments were probably due to differences in the soil environment rather than a result of changes in aboveground processes. Temporal shifts in morphotype frequencies were observed for seedlings in all treatments and indicate that mycorrhizal succession occurred during the study period.     

Impact of ozone, acid mist and soil characteristics on growth and development of fine roots and ectomycorrhiza of young clonal Norway spruce

As part of the joint 14-month exposure experiment on Norway spruce (Picea abies (L.) Karst.) sensitivity to pollution (two levels of ozone plus acid mist) of growth and development of the fine-root system and of mycorrhizae, have been investigated in two forest soils from areas showing forest decline. This study shows that differences in fine-root biomass and the occurrence of species of ectomycorrhizae were mainly due to prevailing conditions within the acid or calcareous substrate. The pollution treatment resulted in higher numbers of short root tips in only one soil, whereas the percentage of ectomycorrhizal roots with a well-developed mantle was low (10-23%) in both soils, irrespective of exposure of trees to ozone and acid mist. There was no consistent response, with the two clones examined, in terms of mycorrhizal frequency, beaded short roots and renewed growth. Regarding the effects on root growth, data cannot be used unrestrictedly for extrapolation to a more complex field situation.     

Mycorrhizal populations and fine root development on Norway spruce exposed to controlled doses of gaseous pollutants and simulated acidic rain treatments

Effects after long-term fumigation and simulated acidic rain (pH 4.0) treatment in open-top chambers (OTC) during 5 vegetation periods on ectomycorrhizal (EM) frequency, fine root structure and distribution of short roots were recorded for Norway spruce. Simultaneous application of ozone and SO(2) resulted in a significant decrease (P< or =0.01) in the proportion of EM root tips and an increase in the numbers of necrotic short roots. The observed differences were partly due to more than additive effects of ozone and SO(2). On the basis of EM and non-mycorrhizal (NM) short roots in the irrigated controls (OTC-V, pH 5.6, Ah-horizon: EM + NM/cm=0.05) the acid treatments resulted in drastic effects (P< or =0.05) on the branching density and the proportion of EM root tips and NM short roots. Relative numbers of short roots infected by indigenous EM fungi with trees in the fumigation treatments at pH 4.0 were similar: 1.5-1.9 EM tips/cm and differed noticeably from the control: 5.2 EM tips/cm. Again, increasing numbers of necrotic and NM short roots were noted. Parallel measurements by the ingrowth-core technique confirm the argument, that EM formation seems to be a reliable indicator of pollution-induced stress.     

90Sr uptake by Pinus ponderosa and Pinus radiata seedlings inoculated with ectomycorrhizal fungi

Strontium-90 ((90)Sr) is a radionuclide characteristic of fallout from nuclear reactor accidents and nuclear weapons testing. Prior studies have shown that Pinus ponderosa and P. radiata seedlings can remove appreciable quantities of (90)Sr from soil and store it in plant tissue. In this study, we inoculated P. ponderosa and P. radiata seedlings with one of five isolates of ectomycorrhizal fungi. Inoculated and noninoculated (control) seedlings were compared for their ability to remove (90)Sr from an organic growth medium. Seedlings were grown in a growth chamber in glass tubes containing 165 cm(3) of sphagnum peat moss and perlite (1 : 1 (v/v)) and, except in the controls, the fungal inoculum. After 3 months, 5978 Bq of (90)Sr in 1 ml of sterile, distilled, deionized water was added. Seedlings were grown for an additional month and then harvested. P. ponderosa seedlings with ectomycorrhizae accumulated 3.0-6.0% of the (90)Sr; bioconcentration ratios (Bq (90)Sr cm(-3) plant tissue/Bq (90)Sr cm(-3) growth medium) ranged from 98-162. Ectomycorrhizal P. radiata seedlings accumulated 6.0-6.9% of the (90)Sr; bioconcentration ratios ranged from 88-133. Nonmycorrhizal P. ponderosa and P. radiata seedlings accumulated only 0.6 and 0.7% of the (90)Sr and had bioconcentration ratios of 28 and 27, respectively. Ectomycorrhizal P. ponderosa and P. radiata seedlings are able to remove 3-5 times more (90)Sr from contaminated soil than seedlings without ectomycorrhizae.     

Effects of simultaneous ozone exposure and nitrogen loads on carbohydrate concentrations, biomass, growth, and nutrient concentrations of young beech trees (Fagus sylvatica)

Beech seedlings were grown under different nitrogen fertilisation regimes (0, 20, 40, and 80 kg Nha(-1)yr(-1)) for three years and were fumigated with either charcoal-filtered (F) or ambient air (O3). Nitrogen fertilisation increased leaf necroses, aphid infestations, and nutrient ratios in the leaves (N:P and N:K), as a result of decreased phosphorus and potassium concentrations. For plant growth, biomass accumulation, and starch concentrations, a positive nitrogen effect was found, but only for fertilisations of up to 40 kg Nha(-1) yr(-1). The highest nitrogen load, however, reduced leaf area, leaf water content, growth, biomass accumulation, and starch concentrations, whereas soluble carbohydrate concentrations were enhanced. The ozone fumigation resulted in reduced leaf area, leaf water content, shoot growth, root biomass accumulation, and decreased starch, phosphorus, and potassium concentrations, increasing the N:P and N:K ratios. A combined effect of the two pollutants was detected for the leaf area and the shoot elongation, where ozone fumigation amplified the nitrogen effects.     

Shift in ectomycorrhizal community composition in Scots pine (Pinus sylvestris L.) seedling roots as a response to nickel deposition and removal of lichen cover

Scots pine seedlings were exposed to wet-deposited nickel (Ni) and removal of lichen cover in a dry heath Scots pine forest. Ni deposition affected the colonization of roots by indigenous ectomycorrhizal fungi in contrasting ways in intact and skimmed quadrats. Highest frequencies of tubercle morphotypes of ectomycorrhiza were found in quadrats exposed to 100 mg m(-2) year(-1) Ni in lichen covered treatment, while in skimmed quadrats these peaked after the treatment with 10 mg Ni m(-2) year(-1). Removal of the lichen layer increased the value of diversity index (H') of ectomycorrhizal fungal community, probably due to the increase in the evenness of the morphotype distribution. Lichen removal seemed also to improve the condition of the short roots, as the frequencies of poor and senescent short roots were decreased by the removal.     

Effects of simultaneous ozone exposure and nitrogen loads on carbohydrate concentrations, biomass, and growth of young spruce trees (Picea abies)

Spruce saplings were grown under different nitrogen fertilization regimes in eight chamberless fumigation systems, which were fumigated with either charcoal-filtered (F) or ambient air (O3). After the third growing season trees were harvested for biomass and non-structural carbohydrate analysis. Nitrogen had an overall positive effect on the investigated plant parameters, resulting in increased shoot elongation, biomass production, fine root soluble carbohydrate concentrations, and also slightly increased starch concentrations of stems and roots. Only needle starch concentrations and fine root sugar alcohol concentrations were decreased. Ozone fumigation resulted in needle discolorations and affected most parameters negatively, including decreased shoot elongation and decreased starch concentrations in roots, stems, and needles. In fine roots, however, soluble carbohydrate concentrations remained unaffected or increased by ozone fumigation. The only significant interaction was an antagonistic effect on root starch concentrations, where higher nitrogen levels alleviated the negative impact of ozone.     

Growth and physiology of Northern Red Oak: preliminary comparisons of mature tree and seedling responses to ozone

Considerable progress has been made during the past decade in the development of mechanistic models that allow complex chemical, physical, and biological processes to be evaluated in the global change context. However, quantitative predictions of the response of individual trees, stands, and forest ecosystems to pollutants and climatic variables require extrapolation of existing data sets, derived largely from seedling studies, to increasing levels of complexity with little or no understanding of the uncertainties associated with these extrapolations. Consequently, a project designed to address concerns associated with scaling from seedling to mature tree responses was initiated. During the 1990 and 1991 growing seasons, mature northern red oak (Quercus rubra L.) trees and seedlings were exposed to subambient, ambient, and twice ambient ozone (O(3)) concentrations. The initial focus of the study was to identify possible trends and obvious differences between mature trees and seedlings, both in terms of growth and physiology and in response to O(3). Generally, mature trees exhibited a greater decrease in photosynthesis rates over the growing season than did the seedlings. Ozone treatments had no consistent effect on gas exchange rates of seedlings, but the twice ambient O(3) treatment resulted in reduced photosynthesis rates in the mature tree. Despite no effect of O(3) on seedling gas exchange rates, total seedling biomass was significantly less at the end of the 1991 growing season for those seedlings exposed to twice ambient O(3) levels. Disproportionate reductions in root biomass also resulted in reduced root to shoot ratios at elevated O(3) concentrations.     

Cd-tolerant Suillus luteus: A fungal insurance for pines exposed to Cd

Soil metal pollution can trigger evolutionary adaptation in soil-borne organisms. An in vitro screening test showed cadmium adaptation in populations of Suillus luteus (L.: Fr.) Roussel, an ectomycorrhizal fungus of pine trees. Cadmium stress was subsequently investigated in Scots pine (Pinus sylvestris L.) seedlings inoculated with a Cd-tolerant S. luteus, isolated from a heavy metal contaminated site, and compared to plants inoculated with a Cd-sensitive isolate from a non-polluted area. A dose-response experiment with mycorrhizal pines showed better plant protection by a Cd-adapted fungus: more fungal biomass and a higher nutrient uptake at high Cd exposure. In addition, less Cd was transferred to aboveground plant parts. Because of the key role of the ectomycorrhizal symbiosis for tree fitness, the evolution of Cd tolerance in an ectomycorrhizal partner such as S. luteus can be of major importance for the establishment of pine forests on Cd-contaminated soils.     

Effect of ectomycorrhizae and ammonium on 134Cs and 85Sr uptake into Picea abies seedlings

Microorganisms play an important role in the fixation of radionuclides in forest soils. In particular, fungi have the capacity to absorb and translocate radionuclides. The role of the ectomycorrhizal fungus Hebeloma crustuliniforme in the uptake of radiocaesium (134Cs) and radiostrontium (85Sr) into seedlings of Norway spruce (Picea abies) was investigated in a pouch test system. Inoculated and non-inoculated seedlings; seedlings inoculated during 8 and 15 weeks; seedlings exposed during 2 and 3 weeks to the radioactive solution; and seedlings grown under low and high ammonium conditions prior to the application of the radionuclides were compared. The final 134Cs and 85Sr activity was determined in fine-roots, main-roots, stems and needles. The results showed that ectomycorrhizae reduced the uptake of 134Cs and 85Sr. The degree of ectomycorrhization was of crucial importance and seemed to be governed by the period during which ectomycorrhizae were allowed to develop and by the ammonium concentration in the nutrient solution. The radionuclide uptake increased with increasing exposure time. Both radionuclides were predominantly accumulated in fine-roots. However, needles proved to describe best the result of net root uptake and translocation to the shoot. The uptake-and and translocation-rates of 85Sr were smaller than those of 134Cs. It is assumed that the translocation is coupled with the intensity of water fluxes through the xylem and that 85Sr is more readily adsorbed into mycelium or plant tissue relative to 134Cs. The effect of high ammonium growth conditions was overcome by the effect of ectomycorrhization, except in needles with a very large biomass which behaved as a strong sink and led to a high accumulation of 134Cs.     

Effects of acid rain on growth and nutrient concentrations in Scots pine and Norway spruce seedlings grown in a nutrient-rich soil

The effects of artificially applied acid precipitation on growth and nutrient concentrations of Scots pine (Pinus sylvestris L.) and Norway spruce (Picea abies [L.] Karst.) seedlings were investigated in a long-term acid irrigation experiment in field conditions. Seedlings of northern and southern origin were planted in boxes containing peat and composted soil rich in nutrients, and sprinkler irrigated with water acidified with nitric and sulphuric acids to pH 3 or pH 4 for periods varying from two to three and a half growing seasons during 1986-1989. Water irrigated (pH 5.4-7.6) and non-irrigated groups of seedlings were also included in the experiment. At the end of the experiment needles, main and lateral shoots and roots were collected from the seedlings for the determination of height growth and biomass partitioning, and for the analysis of S, N, Mg, P, K, Ca, Mn and Fe concentrations. The treatment effects compared to the irrigated control were studied using multivariate analyses of variance and covariance. In the pine seedlings the total dry matter production increased by 25-70% compared with the irrigated controls when the total wet deposition to the seedlings exceeded 67 kg S ha(-1) and 36 kg N ha(-1) (e.g. after two growing seasons' exposure of the pH 3 treatment). The increase was mainly due to an increase in needle dry weight (54-72% greater at pH 3) and root weight (20-65% greater at pH 3), whereas the height growth or shoot weight growth were less affected. The northern provenance pine seedlings responded more clearly to the pH 3 irrigation than the southern ones. The treatments had no consistent effects on any of the growth variables studied in the spruce seedlings, however. The pines had higher root and foliage Ca concentrations as a result of the acid irrigation, whereas in spruce, acid rain decreased the Ca concentration in needles and shoots. Root Mn and Fe concentrations were higher in both species as a result of the pH 3 treatment. A higher soil conductivity and Ca concentration resulted from the prolonged pH 3 treatment. The results strongly support the hypothesis that the long-term growth and nutrient allocation response of conifers to acid precipitation is dependent both on the tree species and on the nutritional status of the soil.     

Changes in the concentrations of phenolics and photosynthates in Scots pine (Pinus sylvestris L.) seedlings exposed to nickel and copper

Studies were done on the effects of elevated soil concentrations of copper (Cu) and (Ni) on foliar carbohydrates and phenolics in Scots pine (Pinus sylvestris L.). Four year-old seedlings were planted in pots filled with metal-treated mineral forest soil in early June. The experimental design included all combinations of four levels of Cu (0, 25, 40 and 50 mg kg(-1) soil dw) and Ni (0, 5, 15 and 25 mg kg(-1) soil dw). Current year needles were sampled for soluble sugar, starch and phenolics at the end of September. Ni increased sucrose concentration in the needles, indicating disturbances in carbohydrate metabolism. Trees exposed to Ni had higher concentrations of condensed tannins compared with controls. In contrast, concentrations of several other phenolic compounds decreased when seedlings were exposed to high levels of Cu or to a combination of Ni and Cu. The results suggest that concentrations of phenolics in Scots pine needles vary in their responses to Ni and Cu in the forest soil.     

Urban polluted forest soils induce elevated root peroxidase activity in Scots pine (Pinus sylvestris L.) seedlings.

Plant biomass. mycorrhizal status and root peroxidase activity were measured in ectomycorrhizal Scots pine (Pinus sylvestris L.) seedlings grown in urban polluted and native, non-polluted forest soils with added ammonium or potassium sulphates simulating N and S deposition of urban areas. Peroxidase activity in the fine roots of seedlings planted in polluted forest soils was higher than in those planted in non-polluted soils and correlated positively with the activities measured in an earlier study in the roots of mature Scots pines growing at the sites from where the soils were collected. Growth of seedlings and mycorrhizal status were not affected by the origin of soil. Exposing the seedlings to winter acclimation conditions for 6 weeks elevated peroxidase activity in the roots. The addition of ammonium or potassium sulphate to non-polluted soils did not induce elevated root peroxidase activity, although at the levels of 0.5 and 1.0 g of ammonium sulphate a slight increasing trend was observed. We suggest, that indirect biotic factors, i.e. changes in the community structure of soil fungi, early stages of recognition, and defence reactions of pine roots against saprophytic and pathogenic fungi may be participating in the elicitation of peroxidase (POD) activity, although the possible role of heavy metals cannot be excluded.     

Interaction of ozone exposure and Fusarium subglutinans inoculation on growth and disease development of loblolly pine seedlings

Seedlings from ten half-sib families of loblolly pine (Pinus taeda) were exposed in open-top chambers to carbon-filtered air (CF), non-filtered air (NF), or air amended with ozone to 1.7 or 2.5 times ambient. After 105 days of exposure, half the seedlings within each family were wounded but not inoculated and half were wounded and inoculated with the pitch canker fungus, Fusarium subglutinans, to which five families were relatively resistant. After an additional 50 days of ozone treatment, seedling growth and canker development were recorded. Cankers were significantly (sigma < or = 0.05) smaller among resistant compared to susceptible families, and were significantly larger among seedlings receiving the highest (2.5) compared to the ambient (NF) ozone treatment. The wound scars of non-inoculated seedlings were also significantly larger among seedlings receiving the 2.5 compared to the NF treatment, but these dimensions did not differ significantly with seedling family or resistance. The weights of needles and large roots were significantly smaller at the 2.5 compared to the 1.7 ozone treatment for inoculated but not for non-inoculated seedlings; this resulted in a significant interaction for ozone and inoculation effects. Among resistant families, root weights were significantly smaller for inoculated seedlings. Diameter growth and dry weights of needles were significantly smaller among inoculated compared to non-inoculated seedlings, but did not differ between NF and 2.5 ozone treatments.     

Compensation processes of Aleppo pine (Pinus halepensis Mill.) to ozone exposure and drought stress

A long-term experiment was performed to study the effects of O3 and drought-stress (DS) on Aleppo pine seedlings (Pinus halepensis Mill.) exposed in open-top chambers. Ozone reduced gas exchange rates, ribulose-1,5-biphosphate carboxylase/oxygenase activity (Rubisco), aboveground C and needle N concentrations and C/N ratio and Ca concentrations of the twigs under 3 mm (twigs<3) and the aerial biomass. Also it increased phosphoenolpyruvate carboxylase (PEPc) and N and K concentrations of the twigs<3. Water stress decreased gas exchange rates, predawn needle water potential (PsiPd), C/N ratio, twigs<3 Ca, plant growth, aerial biomass and increased N, twigs with a diameter above 3 mm P and Mg concentrations. The combined exposure to both stresses increased N concentrations of twigs<3 and roots and aboveground biomass K content and decreased root C, maximum daily assimilation rate and instantaneous water use efficiency. The sensitivity of Aleppo pine to both stresses is determined by plant internal resource allocation and compensation mechanisms to cope with stress.     

Shift in ectomycorrhizal community composition in Scots pine (Pinus sylvestris L.) seedling roots as a response to nickel deposition and removal of lichen cover

Scots pine seedlings were exposed to wet-deposited nickel (Ni) and removal of lichen cover in a dry heath Scots pine forest. Ni deposition affected the colonization of roots by indigenous ectomycorrhizal fungi in contrasting ways in intact and skimmed quadrats. Highest frequencies of tubercle morphotypes of ectomycorrhiza were found in quadrats exposed to 100 mg m⁻² year⁻¹ Ni in lichen covered treatment, while in skimmed quadrats these peaked after the treatment with 10 mg Ni m⁻² year⁻¹. Removal of the lichen layer increased the value of diversity index (H′) of ectomycorrhizal fungal community, probably due to the increase in the evenness of the morphotype distribution. Lichen removal seemed also to improve the condition of the short roots, as the frequencies of poor and senescent short roots were decreased by the removal.     

Conifer aphids in an air-polluted environment. II. Host plant quality

Scots pine (Pinus sylvestris L.) and Norway spruce (Picea abies (L.) Karst.) seedlings were transported to five forest sites at increasing distances from a pulp mill emitting mainly SO(2). Levels of compounds which may have nutritional or defensive value for aphids on pine and spruce seedlings were studied. Glucose and fructose concentrations were significantly increased in pine and spruce needles near the pulp mill. There were no changes in sucrose and starch concentrations. In pine shoots, total free amino acid concentration and the concentrations of ornithine, lysine, histidine and arginine were significantly negatively correlated with the distance from the pulp mill, while in spruce only the individual amino acids glycine, ornithine, lysine and histidine showed a significant negative correlation with distance. There were no changes in total phenolic, catechin, total monoterpene and total resin acid concentrations. However, in pine seedlings monoterpenes beta-pinene and sabinene and in spruce seedlings resin acid palustric acid were significantly correlated with the distance from the pulp mill. The results indicate that SO(2) disturbs carbohydrate metabolism in spruce and pine seedlings. The elevated concentrations of arginine may be the result of the combinations of SO(2), NO(3) and NH(3) emissions of the pulp mill. The emissions did not have any impact on total amounts of defensive substances in trees. Thus, the possible susceptibility of conifers to herbivores appears to be due to changes in nutritive value rather than to reduced chemical resistance.     

Influence of seedling roots, environmental factors and soil characteristics on soil CO2 efflux rates in a 2-year-old loblolly pine (Pinus taeda L.) plantation in the Virginia Piedmont

To understand the role of managed forests in carbon sequestration an understanding of factors controlling soil CO2 efflux will be necessary. This study examined the influence of seedling roots, environmental factors, nutrient availability, and soil characteristics on soil CO2 efflux patterns in a 2-year-old pine plantation in the Virginia Piedmont. Efflux rates were measured both near the base of seedlings and midway between rows in plots that had received fertilization and mulch treatments in a factorial combination. Soil CO2 efflux rates were consistently higher near the base of seedlings, fertilization increased seedling growth with no significant effect on rates. and mulching increased winter efflux rates. In a regression analysis of seasonal soil CO2 efflux, soil temperature explained 42.2% of the variance followed by the interaction of soil temperature and moisture and of soil temperature and plot position, which together explained an additional 9.8% of the observed variance in seasonal rates. During March 2000 measurements, the spatial pattern of soil CO2 efflux between plots was most influenced by differences in soil nitrogen and pine root biomass. Furthermore, spatial differences observed in mean annual efflux rates were found to be highly influenced by the amount of soil coarse fragments in the upper soil profile.