Ethanol synthesis, nitrogen, carbohydrates, and growth in tissues from nitrogen fertilized Pseudotsuga menziesii (Mirb.) Franco and Pinus ponderosa Dougl. ex Laws. seedlings

 Seedlings of Douglas-fir, Pseudotsuga menziesii (Mirb.) Franco, and ponderosa pine, Pinus ponderosa Dougl. ex Laws., were grown in a controlled environment and fertilized with nutrient solutions containing 150 ppm (+N), or 0 ppm nitrogen (−N). These treatments greatly altered seedling growth, and the concentrations of N and carbohydrates in their tissues. Metabolically active tissues, such as roots, incubated with a limited supply of O₂ became hypoxic faster and synthesized more ethanol than less active tissues, such as needles. All tissues that were incubated for 4 h in N₂ synthesized ethanol. Needles incubated in N₂ and light had much lower quantities of ethanol than needles in N₂ and dark, suggesting that O₂ from photosynthetsis limited internal anoxia. Most tissues from +N seedlings synthesized greater quantities of ethanol in N₂ anoxia than tissues from −N seedlings, probably because they were able to produce more enzymes with a greater availability of N. However, this increase in ethanol synthesis between N treatments was most pronounced in the phloem. Ethanol and soluble sugar concentrations were negatively related in needles and positively related in roots of N+ seedlings, but not −N seedlings. Starch concentrations had no effect on the amount of ethanol produced by any tissue. Regardless of N treatments, all tissues from ponderosa pine produced more N₂-induced ethanol than Douglas-fir, in part because its tissues contained different concentrations of soluble sugars and N as a consequence of phenological differences between the species. However, ponderosa pine tissues may also maintain greater quantities of anaerobic enzymes, or their isozymes than Douglas-fir. Received: 22 February 1998 / Accepted 23 June 1998     

Ectomycorrhizal fungi associated with ponderosa pine and Douglas-fir: a comparison of species richness in native western North American forests and Patagonian plantations from Argentina

The putative ectomycorrhizal fungal species registered from sporocarps associated with ponderosa pine and Douglas-fir forests in their natural range distribution (i.e., western Canada, USA, and Mexico) and from plantations in south Argentina and other parts of the world are listed. One hundred and fifty seven taxa are reported for native ponderosa pine forests and 514 taxa for native Douglas-fir forests based on available literature and databases. A small group of genera comprises a high proportion of the species richness for native Douglas-fir (i.e., Cortinarius, Inocybe, and Russula), whereas in native ponderosa pine, the species richness is more evenly distributed among several genera. The comparison between ectomycorrhizal species richness associated with both trees in native forests and in Patagonia (Argentina) shows far fewer species in the latter, with 18 taxa for the ponderosa pine and 15 for the Douglas-fir. Epigeous species richness is clearly dominant in native Douglas-fir, whereas a more balanced relation epigeous/hypogeous richness is observed for native ponderosa pine; a similar trend was observed for Patagonian plantations. Most fungi in Patagonian Douglas-fir plantations have not been recorded in plantations elsewhere, except Suillus lakei and Thelephora terrestris, and only 56% of the fungal taxa recorded in Douglas-fir plantations around the world are known from native forests, the other taxa being new associations for this host, suggesting that new tree + ectomycorrhizal fungal taxa associations are favored in artificial situations as plantations.     

Homeostatic gas-exchange parameters inferred from 13C/12C in tree rings of conifers

The CO₂ concentration of the atmosphere has increased by almost 30% in the past two centuries, with most of the increase (>5 Pa) during the past 60 years. Controlled environment studies of crop plants dependent on the C₃ photosynthetic pathway indicate that an increase of this magnitude would enhance net photosynthesis, reduce stomatal conductance, and increase the difference in CO₂ concentration across the stomata, i.e., CO₂ concentration outside the leaf to that within (c _a-c _i). Here we report evidence, based on stable isotope composition of tree rings from three species of field-grown, native conifer trees, that the trees have indeed responded. However, rather than increasing c _a-c _i, intercellular CO₂ concentrations have shifted upward to match the rise in atmospheric concentrations, holding c _a-c _i constant. No differences were detected among Douglas-fir (Pseudotsuga menziesii), ponderosa pine (Pinus ponderosa), or western white pine (Pinus monticola). The values of c _a-c _i were inferred from stable carbon isotope ratio (¹³C) of tree ring holocellulose adjusted for the 0.6–2.6 difference between holocellulose and whole sapwood. The cellulose extraction removed contaminants deposited in the tree ring after it formed and the adjustment corrected for the enrichment of cellulose relative to whole tissue. The whole sapwood values were then adjusted for bublished estimates of past atmospheric ¹³CO₂ and CO₂ concentrations. To avoid confounding tree age with CO₂, cellulose deposited by saplings in the 1980s was compared to cellulose deposited in the inner rings of nature trees when the mature trees were saplings, between 1910–1929 and 1941–1970; thus saplings were compared to saplings. In a separate analysis, the juvenile effect, which describes the tendency for ¹³C to increase in the first decades of a tree's life, was quantified independent of source CO₂ effects. This study provides evidence that conifers have undergone adjustments in the intercellular CO₂ concentration that have maintained c _a-c _i constant. Based on these results and others, we suggest that c _a-c _i, which has also been referred to as the intrinsic water-use efficiency, should be considered a homeostatic gas-exchange set point for these conifer species.     

Pertilization and nutrition experiments with conifer seedlings in pots

Summary In conifer fertilization and nutrition experimentsPinus halepensis, P. radiata andP. maritima seedlings were grown in pots, filled with soil derived from mica schist and siliceous tertiary deposits and also in peat substrate in paperpots.Fertilization with P ofP. radiata andP. maritima seedlings growing in soil low in available P and N improved seedling height only in combination with N fertilization and fertilization with alone induced P deficiency symptoms. N fertilization with from 100 to 150 ppm (2.4 to 3.2 g N/kg, respectively) in the soil regardless of the form of N (NH₄ ⁺ or NO₃ ^–) applied in the summer or autumn together with application of 20 ppm P before sowing was the fertilization regime which produced the best seedlings.Fertilization of peat before sowingP. halepensis, P. radiata andP. maritima with omission of one of the nutrients N, P and K resulted in visible symptoms of N, P and K deficiency, respectively, in the seedlings. Comparative chemical analysis of needles from the three kinds of conifer seedlings with deficiency symptoms and healthy ones verified the visual symptoms of N and P deficiency but not so convincingly the K deficiency symptoms.     

Impact of Douglas-fir and Scots pine seedlings on plagioclase weathering under acidic conditions

The weathering of soil minerals in forest ecosystems increases nutrient availability for the trees. The rate of such weathering and its relative contribution to forest tree nutrition, is a major issue when evaluating present and potential forest stand productivity and sustainability. The current paper examines the weathering rate of plagioclase with and without Douglas-fir or Scots pine seedlings, in a laboratory experiment at pH 3–4 and 25 °C. All nutrients, with the exception of Ca, were supplied in sufficient amounts in a nutrient solution. The objective of the experiment was to evaluate the potential of trees to mobilise Ca from the mineral plagioclase that contained 12% of Ca. Amounts of nutrients supplied in the nutrient solution, amounts accumulated in the living tissue of the seedlings and amounts leached from the experimental vessels, were measured. A weathering balance, accounting for leached + accumulated − supplied amounts, was established. Bio-induced weathering, defined as the weathering increase in the presence of trees, relative to the weathering rate without trees (geochemical weathering; control vessels), under the present experimental conditions, explained on average, 40% of total weathering (biological + geochemical). These conditions appeared more beneficial to Scots pine (higher relative growth rate, higher Ca incorporation) than to Douglas-fir.     

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.     

Nitrogen metabolism of Douglas fir and Scots pine as affected by optimal nutrition and water supply under conditions of relatively high atmospheric nitrogen deposition

Nitrogen metabolism of the needles of 40-year-old Douglas fir and Scots pine trees, growing in two forest stands on cation-poor and acidic sandy soil with a relatively high atmospheric nitrogen deposition was studied. The composition of the free amino acid (FAA) pool, the concentrations of total nitrogen and soluble protein and the activities of glutamine synthetase (GS) and glutamate dehydrogenase (GDH) were determined in the needles. An excessive nitrogen supply by a high atmospheric nitrogen deposition in both forest stands was indicated by the high concentrations of total nitrogen and the amino acids arginine, glutamic acid, glutamine and aspartic acid in control trees. In addition the effect of optimal nutrition and water supply (fertigation) on the needle nitrogen metabolism was evaluated. The total concentration of the FAA pool in needles of both tree species was lower in the fertigated than in the non-fertigated (control) trees, except for 1-year-old needles of Scots pine, in which the concentration after fertigation did not differ from the control. The lower total FAA concentration in the fertigated trees could be attributed to arginine, the concentration of which was on average 60% lower than in the control. Neither the concentration of soluble protein nor the activity of GS were influenced by fertigation. The activity of GDH in fertigated trees only differed significantly from the control in October. Scots pine needles had higher concentrations of protein (50%) and higher activities of GS (44%) and GDH (25%) than Douglas fir needles. Possible explanations for the lower vitality of Douglas fir compared to Scots pine are given.     

Response of Douglas fir seedlings to phosphorus fertilization and influence of temperature on this response

Summary Effects of P fertilizers on growth of Douglas fir (Pseudotsuga menziesii var.menziesii (Mirb.) Franco.) seedlings were examined in pots and nursery beds. In pot experiments levels of P equivalent to 300 kg/ha were adequate for maximum growth over 14–18 weeks and resulted in available soil P levels of 80 ppm after 15 weeks' growth. Maximum growth in pots was obtained with shoot P concentrations of 0.18%–0.20%, with higher values at lower temperatures, but the optimum concentration for one-year old (1-0) nursery seedlings was 0.16% P. Growth of seedlings was greatly restricted at a soil temperature of 5°C and an air temperature of 12°C. At a soil temperature of 10°C and an air temperature of 14°C seedling P requirement was greater than at soil and air temperatures of 20°C.Comparison showed that monammonium phosphate was more effective than calcium superphosphate in stimulating growth in pots and nursery. Triple superphosphate was also effective in the nursery. Diammonium phosphate, potassium dihydrogen phosphate and phosphoric acid had no advantages as P sources in the nursery. Available P levels of 100–130 ppm, in the loamy sand and sandy loam nurseries studied, and needle P concentrations of 0.18%, when sampled in October, were associated with maximum growth of two-year old (2-0) seedlings.P fertilization decreased root/shoot ratio, but did not alter the allometric relationship of shoot to root. Improving P status from a low level increased root growth capacity in 2-0 seedlings and P fertilization of potted seedlings increased dry weight/height ratio. Uptakes per seed bed ha of 236 kg N, 31 kg P, 81 kg K and 73 kg Ca by 2-0 seedlings were comparable with, or greater than, uptake rates of agricultural crops. Recoveries of 6–11% of P from fertilizer were recorded in the nursery.     

Effect of moisture stress on the growth ofPinus ponderosa Dougl. ex. Laws seedlings in relation to their field performance

Forty wind-pollinated families from eight provenances, four from xeric and four from mesic site types in the Sierra Nevada of California, were examined in the greenhouse for differential response to soil moisture stress. Analysis was made of shoot-root allometric growth coefficients, absolute and relative growth rates and the relationship among these response measures and growth in three field plantations. The results showed significant differences among families for allometric growth (k) coefficients under moderate stress, but no differences among provenances or between site types. Absolute and relative growth rate differences between site types were not significant under any treatment; however provenance differences were observed. Correlation coefficients between plantation performance and greenhouse growth estimates were sometimes significant but significance was plantation and treatment specific. Research performed while author was on sabbatical leave at the Institute of Forest Genetics, USDA-USFS, Placerville, California.     

Effects of past logging and grazing on understory plant communities in a montane Colorado forest

Throughout Pinus ponderosa–Pseudotsuga menziesii forests of the southern Colorado Front Range, USA, intense logging and domestic grazing began at the time of Euro-American settlement in the late 1800s and continued until the early 1900s. We investigated the long-term impacts of these settlement-era activities on understory plant communities by comparing understory composition at a historically logged and grazed site to that of an environmentally similar site which was protected from past use. We found that species richness and cover within functional groups rarely differed between sites in either upland or riparian areas. Multivariate analyses revealed little difference in species composition between sites on uplands, though compositional differences were apparent in riparian zones. Our findings suggest that settlement-era logging and grazing have had only minor long-term impacts on understories of upland Front Range P. ponderosa–P. menziesii forests, though they have had a greater long-term influence on riparian understories, where these activities were likely the most intense. This article was written and prepared by US Government employees on official time, and therefore it is in the public domain and not subject to copyright.     

Influence of induced water stress in ponderosa pine on pine sawflies

Summary Three levels of water stress were induced on pole-size ponderosa pine (Pinus ponderosa) to determine the influence of plant moisture stress on oviposition, survival, and growth of two species of pine sawfly (Neodiprion fulviceps and N. autumnalis). It was found that water stress affected oviposition and subsequent egg survival but not larval development or survival. Stress had a negative effect on early season oviposition (N. fulviceps) and a positive effect on late season oviposition (N. autumnalis). Egg hatch was different between species and years and among treatment levels. Larval development, feeding, and survival were not affected by water stress. Overall, the effect of stress was not sufficient to explain population outbreaks of sawflies. Several hypotheses are presented as possible explanations for the observed experimental results.     

Effect of phosphorus fertilization on water stress in Douglas fir seedlings during soil drying

A growth chamber experiment was conducted to determine if P fertilization to enhance the P nutrition of otherwise N and P deficient Douglas fir [Pseudotsuga menziesii (Mirb.) Franco] seedlings reduces water stress in the seedlings during drought periods. Seedlings were grown in pasteurized mineral soil under well-watered conditions and fertilized periodically with a small amount of nutrient solution containing P at either of three levels: 0, 20, or 50 mg P L^-1. By age 6 mo, leaf nutrient analysis indicated that N and P were deficient in control (0 mg P L^-1) seedlings. The highest level of P fertilization, which doubled leaf P concentration, did not affect plant biomass, suggesting that N deficiency was limiting growth. When these seedlings were subjected to drought, there was no effect of P fertilization on leaf water potential or osmotic potential. Furthermore, P fertilized seedlings had lower stomatal conductance and net photosynthesis rate. These results indicate that enhanced P nutrition, in the presence of N deficiency, does not reduce water stress in Douglas fir seedlings during drought periods.     

Effect of bryophytes on survival of conifer seedlings in subalpine forests of central Japan

Bryophytes form the major seedbed for coniferous trees in the subalpine forests of central Japan. Field experiments were conducted on the floor of a closed stand dominated byTsuga on Mt Fuji to examine the seedling survival ofAbies veitchii andTsuga diversifolia in seven substrate types in relation to the morphological characteristics of the seeds and seedlings. NeitherAbies norTsuga seedlings could survive on dwarf-bamboo litter beyond 2 years after the seed rain.Abies seedlings survived not only in all types of bryophyte communities but also in larch litter. In contrast, the survival ofTsuga seedlings was restricted to communities of smaller-statured bryophytes found on logs. The seeds and 1st year seedlings ofA. veitchii were larger than those ofT. diversifolia. Abies seedlings produced hypocotyls taller than any bryophyte community and radicles which were long enough to penetrate into the moist humus layer, whereasTsuga seedling radicles presumably penetrate into humus only in bryophyte communities forming a thin mat on logs. The effect of bryophytes on survival of tree seedlings therefore depends on the relationships between the morphology of seeds and seedlings and the structure of bryophyte communities.     

Water-soluble organic matter in forest soils

By applying a modified gel permeation technique, the molecular-size distribution (MSD) and complexing properties of water-soluble organic matter (WSOM), isolated from the A_h horizon under stands with either Douglas-fir, European beech or Scots pine were established. Both with respect to MSD and complexing properties, the dissolved organic matter was highly similar. WSOM was comprised of compounds apparently high in molecular weight (>1 kDa) and with a complexing capacity of 1.0±0.1 mol mg^–1 carbon as determined for Cu(II) at pH 5.5 and 0.01 M ionic strength. The effect of WSOM on the partitioning of cations between soil solid phase and soil solution was evaluated in several soil batch experiments using loamy sand or sandy soil material. Although a large part of WSOM was sorbed to the soil matrix, Al, Cu, Fe and Pb were solubilized in considerable amounts by complexation. The Mn concentration in the soil solution was also significantly increased but this probably resulted from a redox reaction, with certain constituents of WSOM serving as electron donor. With a decrease in soil pH, cation mobilization by WSOM was significantly lower as a result of increased sorption and a decrease in complexing capacity of the soluble organics. Application of several low MW aliphatic and phenolic acids gave results similar to the results obtained with WSOM.     

Seasonal variation of growth and development of the roots of five second year conifer species in the nursery

Every other week over their second growing season, stem height, collar diameter, shoot and root dry masses, number of lateral roots and length of the tap root were measured on nursery grown seedlings ofAbies balsamea L. Mill.,Pinus banksiana Lamb.,Pinus resinosa Ait.,Picea mariana Mill. BSP andPicea glauca Moench Voss. Root elongation, branching and mycorrhizal development were also recorded.Given species showed distinct seasonal growth patterns. The rate and timing of maximum root growth (mg/dry weight/week) differed markedly between species.Except for the increase in height ofPinus banksiana, root and shoot growth were not negatively correlated.The results are discussed in relation to the performance of tree seedlings in the nursery.     

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.     

Environmental sensitivity of gas exchange in different-sized trees

The carbon isotope signature (δ¹³C) of foliar cellulose from sunlit tops of trees typically becomes enriched as trees of the same species in similar environments grow taller, indicative of size-related changes in leaf gas exchange. However, direct measurements of gas exchange in common environmental conditions do not always reveal size-related differences, even when there is a distinct size-related trend in δ¹³C of the very foliage used for the gas exchange measurements. Since δ¹³C of foliage predominately reflects gas exchange during spring when carbon is incorporated into leaf cellulose, this implies that gas exchange differences in different-sized trees are most likely to occur in favorable environmental conditions during spring. If gas exchange differs with tree size during wet but not dry conditions, then this further implies that environmental sensitivity of leaf gas exchange varies as a function of tree size. These implications are consistent with theoretical relationships among height, hydraulic conductance and gas exchange. We investigated the environmental sensitivity of gas exchange in different-sized Douglas-fir (Pseudotsuga menziesii) via a detailed process model that specifically incorporates size-related hydraulic conductance [soil–plant–atmosphere (SPA)], and empirical measurements from both wet and dry periods. SPA predicted, and the empirical measurements verified, that differences in gas exchange associated with tree size are greatest in wet and mild environmental conditions and minimal during drought. The results support the hypothesis that annual net carbon assimilation and transpiration of trees are limited by hydraulic capacity as tree size increases, even though at particular points in time there may be no difference in gas exchange between different-sized trees. Maximum net ecosystem exchange occurs in spring in Pacific Northwest forests; therefore, the presence of hydraulic limitations during this period may play a large role in carbon uptake differences with stand-age. The results also imply that the impacts of climate change on the growth and physiology of forest trees will vary depending on the age and size of the forest.