Seasonal changes in amino acids, protein and total nitrogen in needles of fertilized Scots pine trees

Seasonal changes in amino acids, protein and total nitrogen in needles of 30-year-old, fertilized Scots pine (Pinus sylvestris L.) trees growing in Northern Sweden were investigated over two years in field experiments. The studied plots had been fertilized annually for 17 years with (i) a high level of N, (ii) a medium level of N, or (iii) a medium level of N, P and K. Trees growing on unfertilized plots served as controls. In control trees, glutamine, glutamic acid, gamma-aminobutyric acid, aspartic acid and proline represented 50-70% of the total free amino acids determined. Arginine was present only in low concentrations in control trees throughout the year, but it was usually the most abundant amino acid in fertilized trees. Glutamine concentrations were high during the spring and summer in both years of study, whereas proline concentrations were high in the spring but otherwise low throughout the year. In the first year of study, glutamic acid concentrations were high during the spring and summer, whereas gamma-aminobutyric acid was present in high concentrations during the winter months. This pattern was less pronounced in the second year of investigation. The concentrations of most amino acids, except glutamic acid, increased in response to fertilization. Nitrogen fertilization increased the foliar concentration of arginine from < 1 micromol g(dw) (-1) in control trees to a maximum of 110 micromol g(dw) (-1). Trees fertilized with nitrogen, phosphorus and potassium had significantly lower arginine concentrations than trees fertilized with the same amount of nitrogen only. Protein concentrations were similar in all fertilized trees but higher than those in control trees. For all treatments, protein concentrations were high in winter and at a minimum in early spring. In summer, the protein concentration remained almost constant except for a temporary decrease which coincided with the expansion of new shoots. Apart from arginine, the amino acid composition of proteins was similar in all treatments.     

Growth regulation of Scots pine seedlings with different fertilizer compositions and regimes

Scots pine (Pinus sylvestris L.) seedlings were grown in containers filled with peat, using two different fertilizers and three different fertilizer regimes. Seedling shoot and root growth and shoot content of nitrogen, potassium and phosphorus were followed in the nursery and after outplanting in the field. Attempts to regulate growth rate by an exponential nutrient supply were not successful, but the root/shoot ratio was influenced by the fertilization regime. Internal nitrogen concentration was stable only for seedlings with low relative growth rate, while seedlings with high nutrient supply in the nursery showed strong nutrient dilution in the shoot after planting.     

Responses of Scots pine (Pinus sylvestris) seedlings grown in different nutrient regimes to changing root zone temperature in spring

We examined effects of nutrient availability and changing root zone temperature (RZT) on growth, gas exchange and plasma membrane H(+)-ATPase (PM-ATPase) activity of roots of 1-year-old Scots pine (Pinus sylvestris L.) seedlings during spring flushing. The 6-week growth-chamber experiment was carried out in hydroponic cultures that supplied the seedlings with low (0.5 mM N) or high (3 mM N) nutrient concentration and two rates of increase in RZT were simulated: slow warming (SW-treatment) and fast warming (FW-treatment). Air temperature, humidity, and light conditions were similar in all treatments. Growth of roots and shoots was retarded at low RZT, and fresh mass increment of roots was closely correlated with RZT sum. High nutrient availability increased nitrogen concentrations of needles and stems, but only at RZTs >13 degrees C. Low RZT and low availability of nutrients suppressed gas exchange of the seedlings. Real PM-ATPase activity was highly dependent on RZT. At high RZTs, real PM-ATPase activity was affected by nutrient availability but this effect was related to root growth. We conclude that, under conditions of high nutrient availability, Scots pine seedlings can compensate for the suppressive effects of long-term exposure to low RZT by rapidly accelerating growth, gas exchange and root metabolism, but only when RZT has increased above a threshold value, which was 13 degrees C in this study.     

Infection of Scots pine seedlings by Gremmeniella abietina during summer under different inoculum potential

The connection between natural conidia dispersal of Gremmeniella abietina (A‐type) and the disease incidence and severity in first and second year conifer seedlings of Pinus sylvestris was studied in central Finland. The seedlings where exposed to natural infection for a 3‐week period throughout the growth season, followed by a 3‐week incubation period in a growth chamber to promote symptom expression. In second year seedlings the infection periods in May–June had a similar effect as regards disease outbreak and roughly half of the successful infections occurred during these periods. However, the first year seedlings were more severely diseased after the infection period in late July/early August, although the inoculum density was lower during this time compared with the period in late June/early July. This result is consistent with earlier studies. The effect of age and growth phase of the seedlings, temperature sum and the number of conidia on the disease occurrence is discussed and compared with the results of earlier studies in which artificial inoculation was employed.     

Height variation of transplanted seedlings of Scots pine in Finnish nurseries

The height variation of Scots pine (Pinus sylvestris L.) seedling stock was studied using data from annual inventories of seedling crops. The heights of 240 batches of 2-year-old and 3-year-old transplanted seedling from the years 1973–1986 grown at three nurseries in southern Finland were examined. From each batch, 100 clusters of three successive seedlings were systematically located and measured. The mean heights of the seedling batches ranged from 7.8 to 27.0 cm, with a mean of 14.1 cm. Within batch standard deviations ranged from 1.7 to 7.8 cm, with a mean standard deviation of 3.7 cm. There was considerable variation in the mean heights of seedling batches both within a given year and between years. However, estimated variance components of seedling height showed that year, batch and cluster effect each accounted for about 20 percent of the total variance, while seedling effect explained the remaining 40 percent. The implications of results for single tree and batch grading are discussed.     

Nitrate metabolism in Scots pine seedlings during their first growing season

Fluctuations in nitrate reductase activity (NRA), nitrate, nitrite, protein and total nitrogen content of bare-root Scots pine seedlings (Pinus sylvestris L.) raised outdoors were investigated during the first growing season. Nitrate reductase activity was higher in roots than in needles, whereas NO(3) (-) content was higher in needles than in roots and increased in both from June to October. Nitrate reductase activity in roots correlated more closely with NO(3) (-) N in the soil than did NO(3) (-) in the roots. In autumn, there was a closer correlation between foliar NRA and NO(3) (-) in the needles than with NO(3) (-)-N in the soil. Nitrite was not detected in the seedlings during the growing season. Total nitrogen content decreased toward the autumn, whereas protein content initially decreased but increased again in autumn. Acrylic netting placed above the seedlings increased both air and soil temperatures and apparently accelerated the use of nitrate.     

Effect of uninucleate Rhizoctonia on Scots pine and Norway spruce seedlings

One‐year‐old container‐grown seedlings were planted in spring on clear cut areas: the Norway spruce (Picea abies) on a moist upland site (Myrtillus‐type) and Scots pine (Pinus sylvestris) on a dryish upland site (Vaccinium‐type). While still in the nursery, half of the seedlings of each species had been inoculated during the previous summer, with a uninucleate Rhizoctonia sp., a root dieback fungus. At outplanting all the seedlings appeared healthy and had a normal apical bud, although the height of the inoculated seedlings was less than that of the uninoculated control seedlings. At the end of the first growing season after planting, the mortality of inoculated Scots pine and Norway spruce seedlings was 25 and 69%, respectively. After two growing seasons the mortality of inoculated seedlings had increased to 38% for Scots pine and 93% for Norway spruce. The mortality of control seedlings after two growing seasons in the forest was 2% for Scots pine and 13% for Norway spruce. After outplanting the annual growth of inoculated seedlings was poor compared with the growth of control seedlings. These results show that, although Rhizoctonia‐affected seedlings are alive and green in the nursery, the disease subsequently affects both their survival and growth in the forest.     

Leaf- and shoot-level plasticity in response to different nutrient and water availabilities

Phenotypic plasticity in response to environmental variation occurs at all levels of organization and across temporal scales within plants. However, the magnitude and functional significance of plasticity is largely unexplored in perennial species. We measured the plasticity of leaf- and shoot-level physiological, morphological and developmental traits in nursery-grown Populus deltoides Bartr. ex Marsh. individuals subjected to different nutrient and water availabilities. We also examined the extent to which nutrient and water availability influenced the relationships between these traits and productivity. Populus deltoides responded to changes in resource availability with high plasticity in shoot-level traits and moderate plasticity in leaf-level traits. Although shoot-level traits generally correlated strongly with productivity across fertilization and irrigation treatments, few leaf-level traits correlated with productivity, and the relationships depended on the resource examined. In fertilized plants, leaf nitrogen concentration was negatively correlated with productivity, suggesting that growth, rather than enhanced leaf quality, is an important response to fertilization in this species. With the exception of photosynthetic nitrogen-use efficiency, traits associated with resource conservation (leaf senescence rate, water-use efficiency and leaf mass per area) were uncorrelated with short-term productivity in nutrient- and water-stressed plants. Our results suggest that plasticity in shoot-level growth traits has a greater impact on plant productivity than does plasticity in leaf-level traits and that the relationships between traits and productivity are highly resource dependent.     

Morphological and physiological differences in Scots pine seedlings of six seed origins

One-year-old seedlings of Scots pine (Pinus sylvestris L.) offour native seed origins (Loch Maree Islands, Glengarry/GlenMorriston, Glen Affric and Abernethy), a commercial Britishseedlot, and a seedlot from Hedesunda, in middle Sweden, werecompared at monthly intervals from October 1993 to April 1994.Seedling morphology, root condition, root frost hardiness andbud dry matter were determined at each date. There were clear morphological differences among seed origins.Seedlings raised from the commercial seedlot (A70) were largerbut had a poorer root:shoot ratio than the other seed origins.Of the native pines tested, the Loch Maree Islands origin allocateda larger proportion of its photosynthate to fine roots and needlesand smaller proportion to woody structures. Seedlings raised from the commercial British seedlot tendedto have poorer bud lignification than the other origins andalso, in autumn, higher electrolyte leakage rates from its fineroots. During winter, the Swedish origin had the lowest fineroot electrolyte leakage. Seedlings of all origins showed aprogressive increase in fine root hardiness towards mid-winterwith maximum hardiness (–7°C) in January. Dehardeningoccurred over subsequent months reaching –3°C in April.Differences among origins were evident. The Swedish seedlotdeveloped greater frost resistance than the other origins, hardeningbegan earlier in autumn and dehardening began later in spring.The commercial seedlot hardened later than the other originsbut reached a similar level of frost hardiness by January. Ofthe native pines, seedlings of the Loch Maree Islands originwere slowest to develop root hardiness.     

Influence of different nursery container media on rooting of Scots pine and silver birch seedlings after transplanting

Scots pine (Pinus sylvestris L.) and silver birch (Betula pendula Roth.) seedlings were grown in containers filled with growth media based on medium-textured sphagnum peat, coarse perlite and fine sand. The seedlings were then planted into fine and coarse sandy soils in 2.2 l pots, which were subjected to two water-content treatments (only one for birch). After the seedlings had grown five weeks in a greenhouse, rooting into the surrounding soil and shoot growth were measured. Addition of perlite and sand to peat medium slightly affected rooting; thus suggesting minor effects on seedling establishment. However, nitrogen concentration of the seedlings varied between growth media and correlated positively with rooting into the soil. The particle size and water content of the soil affected considerably rooting of the seedlings. Seedling height at the time of planting did not affect rooting or shoot growth. The fact that the fewest out-grown roots occurred in the dry fine sandy soil, suggests that dry soil together with high strength and resistance to root penetration reduce rooting and water uptake by container seedlings most and may thus cause water and nutrient stresses to seedlings after outplanting.     

Growth versus storage: responses of Mediterranean oak seedlings to changes in nutrient and water availabilities

We compare dry mass (DM) and storage of starch (St) and nitrogen (N) in seedlings of three Mediterranean oaks, two evergreens (Quercus coccifera L. and Q. ilex L. subsp. ballota (Desf.) Samp) and one deciduous (Q. faginea Lam.), across different scenarios of nutrient and water availabilities. Three fertilization (5, 50 and 200 mg of N per plant and growing period) and watering (28–39, 55–71 and 70–85 g H₂O 100 g _soil ^?1 gravimetric soil water) treatments were applied to current-year seedlings between May and October 2002 in two independent experiments. The three species showed a similar response to fertilization, storing nitrogen instead of increasing biomass, in agreement with adaptations to nutrient-poor habitats. However, they differed in their responses to water, reflecting the different water requirements in the field: Q. coccifera, from arid zones, showed no response to water regarding DM and St; Q. faginea, from humid zones, required higher water availability to simultaneously increase growth and storage; while Q. ilex, spanning over most of the water availability range, exhibited a balanced increase of both functions when water increased moderately. In the two evergreen species, N concentration increased with water supply, whereas the reverse occurred in Q. faginea. The latter species favoured growth over storage at moderate water supply (according to its more competitive strategy), although it was the species which accumulated more St and N at the end of the experiments (autumn).     

Infection experiments with Gremmeniella abietina on seedlings of Norway spruce and Scots pine

Conidia of Gremmeniella abietina infected and caused disease symptoms in annual shoots of both Scots pine (Pinus sylvestris) and Norway spruce (Picea abies) seedlings. In Norway spruce shoots the infection remained largely latent, with only a few seedlings showing symptoms. Mycelial growth inside the shoots was faster in Scots pine than in Norway spruce and was favoured by low temperature in both hosts. The shoots of Norway spruce seedlings had higher endophyte populations than those of Scots pine, and the populations were decreased by low temperatures. Reductions in the normal epiphytic or endophytic flora by acid mist treatments seemed to favour the development of G. abietina.     

Survival of Scots pine seedlings after biological and chemical control of damping-off in plastic greenhouses

Survival of pine seedlings was tested in greenhouses where substrates were infected with Rbizoctonia, Fusarium or Rbizoctonia with Fusarium. To reduce the pathogen menace biological (sterile sawdust, Tricboderma or sawdust with Tricboderma) or chemical (seed protection) treatments were performed. Incorporation of sawdust or sawdust with Tricboderma 20 days before sowing increased the survival of seedlings and it was comparable to protection with Nemispor for Fusarium-menace, with Funaben T for Rbizoctonia menace and Oxafun T or Karbochin for both pathogens.     

Regulation of organic and inorganic nitrogen uptake in Scots pine (Pinus sylvestris) seedlings

Plants possess regulatory mechanisms that enhance nitrogen (N) uptake under conditions of spatial and temporal variation in N availability. Study of regulatory mechanisms has focused almost exclusively on the uptake of inorganic N sources (i.e., ammonium (NH4+), nitrate (NO3-). Several lines of evidence, however, suggest that amino acids may constitute a potential source of N for a number of plant species, including conifers. In the present study, we investigated the uptake of amino acids and inorganic N in Scots pine (Pinus sylvestris L.) seedlings grown at different N concentrations. We compared the uptake rate of the individual N sources using U-[13C2], [15N]-glycine, U-[13C6], [15N4]-arginine, 15NH4, or 15NO3, and tested the short-term effect of N supply on the uptake rate of glycine, arginine and in field-grown Scots pine seedlings. Our data indicate that Scots pine seedlings can absorb substantial amounts of N in the form of intact arginine and glycine molecules. The data also suggest that Scots pine seedlings down-regulate their uptake of NH4+-N and arginine-N, but not of glycine-N in response to increased endogenous N concentrations.     

Changes in nitrogen level and mycorrhizal structure of Scots pine seedlings inoculated with Thelephora terrestris

? We investigated the influence of Thelephora terrestris, an ectomycorrhizal fungus, on the concentration of nitrogen in needles of Pinus sylvestris L. seedlings and their mycorrhizal structure within four seasons of growth.

? Seedlings were grown in four treatments: (I) inoculated + 0.030 g N, (II) inoculated + 0.045 g N, (III non-inoculated + 0.030 g N, (IV) non-inoculated + 0.045 g N under laboratory conditions and later planted in post-agricultural land.

? For inoculated treatments, statistically significant differences in N concentration of needles were observed for two- and four-year-old seedlings. The number of ectomycorrhizae and concentration of N in needles were negatively correlated. After four seasons of growth the greatest numbers of ectomycorrhizae were present on seedlings from treatment II, and the smallest on seedlings from treatment IV.

? Two-year-old seedlings had been colonized by at least six different fungal taxa, i.e. Cenococcum geophilum, Hebeloma crustuliniforme, Rhizopogon sp., Suillus bovinus, Thelephora terrestris and a non-identified one. Treatments I and II were dominated by the ectomycorrhizae of T. terrestris and their number was significantly greater than in treatments III or TV. On four-year-old seedlings ectomycorrhizae of T. terrestris were dominated in all treatments, while ectomycorrhizae of Hebeloma sp. were not found.

     

Growth patterns from different forest generations of Scots pine in Estonia

There is strong evidence that climate change alters tree growth in boreal forests. In Estonia, the analysis of ring measurements has been a common method to study growth changes. In this study, annual height growth data from dominant or co-dominant Scots pine (Pinus sylvestris L.) trees were used to develop a growth model for three forest generations. Stem analysis was applied and annual height growth was measured as the distance between whorls, as detected by branch knots of whorls on the split stem surface. Retrospective analysis of height growth produced comparative trends for three different age groups. Statistical analyses were used to estimate the impact of different factors on growth. Annual height growth was considered the best indicator for detecting possible trends in the growth potential of trees. Study results indicate that three generations (separated by time periods of 30–40 years) showed significant differences in growth patterns caused by shifts in climatic factors and management regimes (anthropogenic and natural disturbances).     

Relationship between temperature, respiratory loss of sugar and premature dehardening in dormant Scots pine seedlings

Increased intracellular sugar concentration is an important contributor to the increased cold tolerance of conifers in winter. This study examines the extent to which wintertime respiratory loss of sugars leads to premature dehardening. Two-year-old seedlings of Scots pine (Pinus sylvestris L.), grown and cold-hardened in the field, were exposed to different temperature regimes for 16 weeks while dormant. To minimize short-term carry-over effects, after the temperature treatments, all seedlings were conditioned to 5.5 degrees C and watered before the assessment of non-structural carbohydrates and cold tolerance. Needle sugar concentration was decreased by 54, 32, 21 and 9% following treatment at 5.5, 0, -1.5 and -8.5 degrees C, respectively. Sugar concentration did not decrease as much in root tissues as in needles because starch was mobilized in roots. Cold tolerance of needles was analyzed by controlled freezing, and the temperature causing an initial 10% damage (LT(10)) was plotted as a function of needle sugar concentration, revealing a strong, linear relationship. When one-third of the initial sugars had been consumed, LT(10) had increased from -24.5 to -16.5 degrees C, and when one half had been consumed, LT(10) had increased to -10 degrees C. Consequences of these findings for the field performance of conifers are discussed in relation to climatic variation and change.     

Inter-specific competition and management modify the morphology, nutrient content and resorption in Scots pine needles

Influence of tree-to-tree competition on nutrient resorption is still not well understood. To contribute filling this gap, we assessed the effects of thinning (0, 20 and 30 % extraction of basal area) and canopy type (beech–pine vs. pine subplots) on needle dry weight, needle length, nutrient content and nutrient resorption (N, P and K) in Scots pine (Pinus sylvestris) needles of different cohorts, 8–9 years after thinning. Thinning and canopy type often concurrently affected needle morphology (e.g. lighter and shorter needles in 30 % thinning, heavier and longer needles in pine canopy on the first year of study) and nutrient content (e.g. decrease in N, P and K in 30 %; N, P and K higher in pine canopy on the first year of study). However, effects of thinning appeared only in older cohorts for N and P but were found in old and new cohorts for needle dry weight, needle length and K, indicating that some thinning effects remained after 8–9 years. Canopy effects on morphology and nutrient content were more frequent in recent cohorts, in relation to an increase in beech cover over time. While no clear effects of thinning on the nutrient resorption were observed, higher values were observed in the pine than in the mixed canopy, which could be related to a higher Scots pine stem growth in those patches. The observed differences between treatments will likely increase as the stand develops, probably leading to beech trees being dominant, and as future thinnings are carried out.     

Diversity of ophiostomatoid fungi associated with the large pine weevil, Hylobius abietis, and infested Scots pine seedlings in Poland

Context

Bark beetles are known to be associated with fungi, especially the ophiostomatoid fungi. However, very little is known about role of pine weevils, e.g., Hylobius abietis, as a vector of these fungi in Europe.

Aims

The aims of our study were to demonstrate the effectiveness of H. abietis as a vector of ophiostomatoid fungi in Poland and to identify these fungi in Scots pine seedlings damaged by weevil maturation feeding.

Methods

Insects and damaged Scots pine seedlings were collected from 21 reforestation sites in Poland. The fungi were identified based on morphology, DNA sequence comparisons for two gene regions (ITS, β-tubulin) and phylogenetic analyses.

Results

Sixteen of the ophiostomatoid species were isolated and identified. In all insect populations, Leptographium procerum was the most commonly isolated fungus (84 %). Ophiostoma quercus was also found at a relatively high frequency (16 %). Other ophiostomatoid fungi were found only rarely. Among these rarer fungi, four species, Leptographium lundbergii, Ophiostoma floccosum, Ophiostoma piliferum and Sporothrix inflata, were isolated above 3 %. L. procerum was isolated most frequently and was found in 88 % of the damaged seedlings. S. inflata was isolated from 26 %, while O. quercus occurred in 10 % of the seedlings.

Conclusion

This study confirmed that L. procerum and O. quercus were common associates of H. abietis, while others species were found inconsistently and in low numbers, indicating causal associations. H. abietis also acted as an effective vector transmitting ophiostomatoid species, especially L. procerum and S. inflata, to Scots pine seedlings.