Within-crown plasticity in leaf traits among the tallest conifers.

PREMISE OF THE STUDY: Leaves are the sites of greatest water stress in trees and a key means of acclimation to the environment. We considered phenotypic plasticity of Pseudotsuga menziesii leaves in their ecological context, exploring responsiveness to natural gradients in water stress (indicated by sample height) and light availability (measured from hemispherical photos) to understand how leaf structure is controlled by abiotic factors in tall tree crowns. METHODS: After measuring anatomy, morphology, and carbon isotope composition (δ13 C) of leaves throughout crowns of P. menziesii >90 m tall, we compared structural plasticity of leaves among the three tallest conifer species using equivalent data from past work on Sequoia sempervirens and Picea sitchensis. KEY RESULTS: Leaf mass per projected area (LMA) and δ13 C increased and mesoporosity (airspace/area) decreased along the water-stress gradient, while light did not play a detectable role in leaf development. Overall, leaves of P. menziesii were far less phenotypically responsive to within-crown abiotic gradients than either P. sitchensis, whose leaves responded strongly to light availability, or S. sempervirens, whose leaves responded equally strongly to water stress. CONCLUSIONS: P. menziesii maintain remarkably consistent leaf structure despite pronounced vertical gradients in abiotic factors. Contrasting patterns of leaf structural plasticity underlie divergent ecological strategies of the three tallest conifer species, which coexist in Californian rainforests.

Resin duct size and density as ecophysiological traits in fire scars of Pseudotsuga menziesii and Larix occidentalis.

BACKGROUND AND AIMS: Resin ducts (RDs) are features present in most conifer species as defence structures against pests and pathogens; however, little is known about RD expression in trees following fire injury. This study investigates changes in RD size and density in fire scars of Douglas fir (Pseudotsuga menziesii) and western larch (Larix occidentalis) as a means to evaluate the ecophysiological significance of traumatic resinosis for tree defence and survival. METHODS: Transverse and tangential microsections were prepared for light microscopy and image analysis in order to analyse axial and radial RDs, respectively. Epithelial cells of RDs and fusiform rays associated with radial RDs were also examined. RDs were compared between normal xylem and wound xylem at four different section heights along the fire-injured stem. KEY RESULTS: Following fire injury, P. menziesii axial RDs narrowed by 38-43 % in the first year after injury, and the magnitude of this change increased with stem height. Larix occidentalis axial RDs widened by 46-50 % in the second year after injury. Radial RDs were of equivalent size in P. menziesii, but widened by 162-214 % in L. occidentalis. Fusiform rays were larger following fire injury, by 4-14 % in P. menziesii and by 23-38 % in L. occidentalis. Furthermore, axial RD density increased in both species due to the formation of tangential rows of traumatic RDs, especially in the first and second years after injury. However, radial RD density did not change significantly. CONCLUSIONS: These results highlight traumatic resinosis as a species-specific response. Pseudotsuga menziesii produce RDs of equivalent or reduced size, whereas L. occidentalis produce wider RDs in both the axial and radial duct system, thereby increasing resin biosynthesis and accumulation within the whole tree. Larix occidentalis thus appears to allocate more energy to defence than P. menziesii.

Tree-ring stable isotopes record the impact of a foliar fungal pathogen on CO(2) assimilation and growth in Douglas-fir.

Swiss needle cast (SNC) is a fungal disease of Douglas-fir (Pseudotsuga menziesii) that has recently become prevalent in coastal areas of the Pacific Northwest. We used growth measurements and stable isotopes of carbon and oxygen in tree-rings of Douglas-fir and a non-susceptible reference species (western hemlock, Tsuga heterophylla) to evaluate their use as proxies for variation in past SNC infection, particularly in relation to potential explanatory climate factors. We sampled trees from an Oregon site where a fungicide trial took place from 1996 to 2000, which enabled the comparison of stable isotope values between trees with and without disease. Carbon stable isotope discrimination (Δ(13)C) of treated Douglas-fir tree-rings was greater than that of untreated Douglas-fir tree-rings during the fungicide treatment period. Both annual growth and tree-ring Δ(13)C increased with treatment such that treated Douglas-fir had values similar to co-occurring western hemlock during the treatment period. There was no difference in the tree-ring oxygen stable isotope ratio between treated and untreated Douglas-fir. Tree-ring Δ(13)C of diseased Douglas-fir was negatively correlated with relative humidity during the two previous summers, consistent with increased leaf colonization by SNC under high humidity conditions that leads to greater disease severity in following years.

The impacts of water stress on phloem transport in Douglas-fir trees.

Despite the critical role that phloem plays in a number of plant functional processes and the potential impact of water stress on phloem structural and phloem sap compositional characteristics, little research has been done to examine how water stress influences phloem transport. The objectives of this study were to develop a more accurate understanding of how water stress affects phloem transport in trees, both in terms of the short-term impacts of water stress on phloem sap composition and the longer-term impacts on sieve cell anatomical characteristics. Phloem sieve cell conductivity (kp) was evaluated along a gradient of tree height and xylem water potential in Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) trees in order to evaluate the influence of water stress on phloem transport capacity. The Hagen-Poiseuille equation was used with measurements of sieve cell anatomical characteristics, water content of phloem sap, non-structural carbohydrate content of phloem sap and shoot water potential (Ψl) to evaluate impacts of water stress on kp. Based on regression analysis, for each 1 MPa decrease in mean midday Ψl, sieve cell lumen radius decreased by 2.63 µm MPa(-1). Although there was no significant trend in sucrose content with decreasing Ψl, glucose and fructose content increased significantly with water stress and sieve cell relative water content decreased by 13.5% MPa(-1), leading to a significant increase in sugar molar concentration of 0.46 mol l(-1) MPa(-1) and a significant increase in viscosity of 0.27 mPa s MPa(-1). Modeled kp was significantly influenced both by trends in viscosity as well as by water stress-related trends in sieve cell anatomy.

Do ray cells provide a pathway for radial water movement in the stems of conifer trees?

PREMISE OF THE STUDY: The pathway of radial water movement in tree stems presents an unknown with respect to whole-tree hydraulics. Radial profiles have shown substantial axial sap flow in deeper layers of sapwood (that may lack direct connection to transpiring leaves), which suggests the existence of a radial pathway for water movement. Rays in tree stems include ray tracheids and/or ray parenchyma cells and may offer such a pathway for radial water transport. This study investigated relationships between radial hydraulic conductivity (k(s-rad)) and ray anatomical and stem morphological characteristics in the stems of three conifer species whose distributions span a natural aridity gradient across the Cascade Mountain range in Oregon, United States. METHODS: The k(s-rad) was measured with a high-pressure flow meter. Ray tracheid and ray parenchyma characteristics and water transport properties were visualized using autofluorescence or confocal microscopy. KEY RESULTS: The k(s-rad) did not vary predictably with sapwood depth among species and populations. Dye tracer did not infiltrate ray tracheids, and infiltration into ray parenchyma was limited. Regression analyses revealed inconsistent relationships between k(s-rad) and selected anatomical or growth characteristics when ecotypes were analyzed individually and weak relationships between k(s-rad) and these characteristics when data were pooled by tree species. CONCLUSIONS: The lack of significant relationships between k(s-rad) and the ray and stem morphologies we studied, combined with the absence of dye tracer in ray tracheid and limited movement of dye into ray parenchyma suggests that rays may not facilitate radial water transport in the three conifer species studied.

Water stress, shoot growth and storage of non-structural carbohydrates along a tree height gradient in a tall conifer.

We analysed concentrations of starch, sucrose, glucose and fructose in upper branch wood, foliage and trunk sapwood of Douglas-fir trees in height classes ranging from ~2 to ~57 m. Mean concentrations of non-structural carbohydrates (NSC) for all tissues were highest in the tallest height class and lowest in the lowest height class, and height-related trends in NSC were most pronounced in branches. Throughout a 17-month sampling period, mean values of branch NSC from the 57 m trees ranged between 30 and 377% greater than the 2 m trees. Branch NSC was inversely correlated with midday shoot water potential (Ψ(l)), shoot osmotic potential at full turgor (Ψ) and shoot extension. Temporal fluctuation in branch NSC was inversely correlated with height, and positively correlated with midday Ψ(l) , Ψ and shoot extension. The positive correlation between height and storage of NSC, and the negative correlation between NSC storage and shoot extension provide evidence that size-related growth decline in trees is not strongly associated with constraints on photosynthesis. The negative correlation between height and fluctuation in NSC suggests that mobilization of photosynthate in taller trees is constrained by some factor such as reductions in turgor-driven cell expansion or constraints on phloem transport.

Defining how aging Pseudotsuga and Abies compensate for multiple stresses through multi-criteria assessment of a functional-structural model.

Many hypotheses have been advanced about factors that control tree longevity. We use a simulation model with multi-criteria optimization and Pareto optimality to determine branch morphologies in the Pinaceae that minimize the effect of growth limitations due to water stress while simultaneously maximizing carbohydrate gain. Two distinct branch morphologies in the Pareto optimal space resemble Pseudotsuga menziesii (Mirb.) Franco and Abies grandis (Dougl. ex D. Don) Lindl., respectively. These morphologies are distinguished by their performance with respect to two pathways of compensation for hydraulic limitation: minimizing the mean path length to terminal foliage (Pseudotsuga) and minimizing the mean number of junction constrictions to terminal foliage (Abies). Within these two groups, we find trade-offs between the criteria for foliage display and the criteria for hydraulic functioning, which shows that an appropriate framework for considering tree longevity is how trees compensate, simultaneously, for multiple stresses. The diverse morphologies that are found in a typical old-growth conifer forest may achieve compensation in different ways. The method of Pareto optimization that we employ preserves all solutions that are successful in achieving different combinations of criteria. The model for branch development that we use simulates the process of delayed adaptive reiteration (DAR), whereby new foliage grows from suppressed buds within the established branch structure. We propose a theoretical synthesis for the role of morphology in the persistence of old Pseudotsuga based on the characteristics of branch morphogenesis found in branches simulated from the optimal set. (i) The primary constraint on branch growth for Pseudotsuga is the mean path length; (ii) as has been previously noted, DAR is an opportunistic architecture; and (iii) DAR is limited by the number of successive reiterations that can form. We show that Pseudotsuga morphology is not the only solution to old-growth constraints, and we suggest how the model results should be used to guide future empirical investigation based on the two contrasting morphologies and how the morphological contrast may relate to physiological processes. Our results show that multi-criteria optimization with Pareto optimality has promise to advance the use of models in theory development and in exploration of functional-structural trade-offs, particularly in complex biological systems with multiple limiting factors.

Height-related trends in stomatal sensitivity to leaf-to-air vapour pressure deficit in a tall conifer.

Stomatal responses to leaf-to-air vapour pressure deficit (LVPD), leaf water potential components, and cuticular properties were characterized for Douglas-fir (Pseudotsuga menziesii) foliage collected from tree tops along a height gradient from 5 m to 58 m in order to explore height-related trends in stomatal sensitivity to LVPD and to investigate the role of bulk leaf turgor and leaf cuticle thickness in determining stomatal behaviour. There were three distinct phases in the response of stomatal conductance (g(s)) to changes in LVPD. At low LVPD, g(s) increased with increasing LVPD (phase one). During the second phase, g(s) was maximal at low to intermediate LVPD and during the third phase g(s) declined steadily as LVPD increased. The responsiveness of g(s) to LVPD exhibited a height-related pattern such that maximum g(s) (g(s-max)) occurred at progressively greater LVPD with increasing height (r2=0.55, P=0.006). Bulk leaf osmotic potential at full turgor decreased with height (r2=0.77, P=0.00016), and LVPD at g(s-max) and at maximum crown conductance (g(c-max)) in the field were significantly correlated with leaf turgor (r2=0.92, P=0.0093). Cuticle thickness increased by 0.044 microm for every metre increase in height (r2=0.78, P=0.00015). The observed trends in the response of g(s) to LVPD along a height gradient, and their consistency with height-related trends in foliar osmotic potential suggest that osmotic adjustment at the tops of tall trees influences the relationship between g(s) and LVPD.

Architecture of the wood-wide web: Rhizopogon spp. genets link multiple Douglas-fir cohorts.

*The role of mycorrhizal networks in forest dynamics is poorly understood because of the elusiveness of their spatial structure. We mapped the belowground distribution of the fungi Rhizopogon vesiculosus and Rhizopogon vinicolor and interior Douglas-fir trees (Pseudotsuga menziesii var. glauca) to determine the architecture of a mycorrhizal network in a multi-aged old-growth forest. *Rhizopogon spp. mycorrhizas were collected within a 30 x 30 m plot. Trees and fungal genets were identified using multi-locus microsatellite DNA analysis. Tree genotypes from mycorrhizas were matched to reference trees aboveground. Two trees were considered linked if they shared the same fungal genet(s). *The two Rhizopogon species each formed 13-14 genets, each colonizing up to 19 trees in the plot. Rhizopogon vesiculosus genets were larger, occurred at greater depths, and linked more trees than genets of R. vinicolor. Multiple tree cohorts were linked, with young saplings established within the mycorrhizal network of Douglas-fir veterans. A strong positive relationship was found between tree size and connectivity, resulting in a scale-free network architecture with small-world properties. *This mycorrhizal network architecture suggests an efficient and robust network, where large trees play a foundational role in facilitating conspecific regeneration and stabilizing the ecosystem.

Coordination of leaf structure and gas exchange along a height gradient in a tall conifer.

The gravitational component of water potential and frictional resistance during transpiration lead to substantial reductions in leaf water potential (Psi(l)) near the tops of tall trees, which can influence both leaf growth and physiology. We examined the relationships between morphological features and gas exchange in foliage collected near the tops of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) trees of different height classes ranging from 5 to 55 m. This sampling allowed us to investigate the effects of tree height on leaf structural characteristics in the absence of potentially confounding factors such as irradiance, temperature, relative humidity and branch length. The use of cut foliage for measurement of intrinsic gas-exchange characteristics allowed identification of height-related trends without the immediate influences of path length and gravity. Stomatal density, needle length, needle width and needle area declined with increasing tree height by 0.70 mm(-2) m(-1), 0.20 mm m(-1), 5.9 x 10(-3) mm m(-1) and 0.012 mm(2) m(-1), respectively. Needle thickness and mesophyll thickness increased with tree height by 4.8 x 10(-2) mm m(-1) and 0.74 microm m(-1), respectively. Mesophyll conductance (g(m)) and CO(2) assimilation in ambient [CO(2)] (A(amb)) decreased by 1.1 mmol m(-2) s(-1) per m and 0.082 micromol m(-2) s(-1) per m increase in height, respectively. Mean reductions in g(m) and A(amb) of foliage from 5 to 55 m were 47% and 42%, respectively. The observed trend in A(amb) was associated with g(m) and several leaf anatomic characteristics that are likely to be determined by the prevailing vertical tension gradient during foliar development. A linear increase in foliar delta(13)C values with height (0.042 per thousand m(-1)) implied that relative stomatal and mesophyll limitations of photosynthesis in intact shoots increased with height. These data suggest that increasing height leads to both fixed structural constraints on leaf gas exchange and dynamic constraints related to prevailing stomatal behavior.

Height-related trends in leaf xylem anatomy and shoot hydraulic characteristics in a tall conifer: safety versus efficiency in water transport.

Hydraulic vulnerability of Douglas-fir (Pseudotsuga menziesii) branchlets decreases with height, allowing shoots at greater height to maintain hydraulic conductance (K shoot) at more negative leaf water potentials (Psi l). To determine the basis for this trend shoot hydraulic and tracheid anatomical properties of foliage from the tops of Douglas-fir trees were analysed along a height gradient from 5 to 55 m. Values of Psi l at which K shoot was substantially reduced, declined with height by 0.012 Mpa m(-1). Maximum K shoot was reduced by 0.082 mmol m(-2) MPa(-1) s(-1) for every 1 m increase in height. Total tracheid lumen area per needle cross-section, hydraulic mean diameter of leaf tracheid lumens, total number of tracheids per needle cross-section and leaf tracheid length decreased with height by 18.4 microm(2) m(-1), 0.029 microm m(-1), 0.42 m(-1) and 5.3 microm m(-1), respectively. Tracheid thickness-to-span ratio (tw/b)2 increased with height by 1.04 x 10(-3) m(-1) and pit number per tracheid decreased with height by 0.07 m(-1). Leaf anatomical adjustments that enhanced the ability to cope with vertical gradients of increasing xylem tension were attained at the expense of reduced water transport capacity and efficiency, possibly contributing to height-related decline in growth of Douglas fir.

Partitioning variation in Douglas-fir xylem properties among multiple scales via a Bayesian hierarchical model.

Hierarchical biological scales permeate research in tree physiology and represent multiple sources of variation. We discuss the importance of matching the sampling and analysis scales to biological scales in the data. The advantages of statistical hierarchical modeling are demonstrated using the relationship between specific conductivity and tracheid diameter of secondary xylem as an example. The structure and results of three statistical models were compared within a Bayesian context: a simple linear regression (SLR); a repeated measures analysis (REP); and a hierarchical model (HM). The models share similar mean structures but differ in how variation is partitioned among scales: the SLR model assumes independence among observations (variation came from only a single scale); the REP allows multiple observations of each tree to be correlated; and the HM incorporates features of the REP with an additional variance structure that partitions variation across a broader scale. Our data included hierarchical scales of position on the tree, tree, fertilization treatment and species (Pseudotsuga menziesii (Mirb.) Franco). The HM gave more precise estimates for model parameters, was more robust to outliers, provided a more detailed description of covariances within the data at multiple scales compared with the SLR and REP and increased our ability to detect differences among positions on the tree. The proper statistical analyses increase the value of research by allowing the most exact interpretation.

Biophysical constraints on leaf expansion in a tall conifer.

The physiological mechanisms responsible for reduced extension growth as trees increase in height remain elusive. We evaluated biophysical constraints on leaf expansion in old-growth Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) trees. Needle elongation rates, plastic and elastic extensibility, bulk leaf water (Psi(L)) and osmotic (Psi(pi)) potential, bulk tissue yield threshold and final needle length were characterized along a height gradient in crowns of > 50-m-tall trees during the period between bud break and full expansion (May to June). Although needle length decreased with increasing height, there was no height-related trend in leaf plastic extensibility, which was highest immediately after bud break (2.9%) and declined rapidly to a stable minimum value (0.3%) over a 3-week period during which leaf expansion was completed. There was a significant positive linear relationship between needle elongation rates and plastic extensibility. Yield thresholds were consistently lower at the upper and middle crown sampling heights. The mean yield threshold across all sampling heights was 0.12 +/- 0.03 MPa on June 8, rising to 0.34 +/- 0.03 MPa on June 15 and 0.45 +/- 0.05 MPa on June 24. Bulk leaf Psi(pi) decreased linearly with increasing height at a rate of 0.004 MPa m(-1) during the period of most rapid needle elongation, but the vertical osmotic gradient was not sufficient to fully compensate for the 0.015 MPa m(-1) vertical gradient in Psi(L), implying that bulk leaf turgor declined at a rate of about 0.011 MPa m(-1) increase in height. Although height-dependent reductions in turgor appeared to constrain leaf expansion, it is possible that the impact of reduced turgor was mitigated by delayed phenological development with increasing height, which resulted in an increase with height in the temperature during leaf expansion.

Hydraulic architecture and photosynthetic capacity as constraints on release from suppression in Douglas-fir and western hemlock.

We compared hydraulic architecture, photosynthesis and growth in Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco), a shade-intolerant species, and western hemlock (Tsuga heterophylla (Raf.) Sarg.), a shade-tolerant species, to study the temporal pattern of release from suppressive shade. In particular, we sought to determine whether hydraulic architecture or photosynthetic capacity is most important in constraining release. The study was conducted at two sites with mixed stands of 10- to 20-year-old Douglas-fir and western hemlock. At one site, the stand had been thinned allowing release of the understory trees, whereas at the other site, the stand remained unthinned. Douglas-fir had lower height growth (from 1998-2003) and lower relative height growth (height growth from 1998 to 2003/height in 1998) than western hemlock. However, relative height growth of released versus suppressed trees was higher in Douglas-fir (130%) than in western hemlock (65%), indicating that, although absolute height growth was less, Douglas-fir did release from suppression. Release seemed to be constrained initially by a limited photosynthetic capacity in both species. Five years after release, Douglas-fir trees had 14 times the leaf area and 1.5 times the leaf nitrogen concentration (N (area)) of suppressed trees. Needles of released western hemlock trees had about twice the maximum assimilation rate (A (max)) at ambient [CO(2)] as needles of suppressed trees and exhibited no photoinhibition at the highest irradiances. After release, trees increased in leaf area, leaf N concentration and overall photosynthetic capacity. Subsequently, hydraulic architecture appeared to constrain release in Douglas-fir and, to a lesser extent, in western hemlock. Released trees had significantly less negative foliar delta(13)C values than suppressed trees and showed a positive relationship between leaf area:sapwood area ratio (A (L)/A (S)) and delta(13)C, suggesting that trees with more leaf area for a given sapwood area experienced a stomatal limitation on carbon gain. Nonetheless, these changes had no significant effects on leaf specific conductivities of suppressed versus released trees of either species, but leaf specific root conductance was significantly lower in released Douglas-fir.

Distinct developmental mechanisms reflect the independent origins of leaves in vascular plants.

Vascular plants diverged more than 400 million years ago into two lineages, the lycophytes and the euphyllophytes . Leaf-like organs evolved independently in these two groups . Microphylls in lycophytes are hypothesized to have originated as lateral outgrowths of tissue that later became vascularized (the enation theory) or through the sterilization of sporangia (the sterilization hypothesis) . Megaphylls in euphyllophytes are thought to represent modified lateral branches . The fossil record also indicates that the seed plant megaphyll evolved uniquely in the ancestor of seed plants, independent of megaphylls in ferns, because seed plants evolved from leafless progymnosperm ancestors . Surprisingly, a recent study of KNOX and ARP gene expression in a lycophyte was reported to indicate recruitment of a similar mechanism for determinacy in both types of leaves . We examined the expression of Class III HD-Zip genes in the lycophyte Selaginella kraussiana and in two gymnosperms, Ginkgo and Pseudotsuga. Our data indicate that mechanisms promoting leaf initiation, vascularization, and polarity are quite different in lycophytes and seed plants, consistent with the hypotheses that megaphylls originated as lateral branches whereas microphylls originated as tissue outgrowths.

Uptake and translocation of manganese in seedlings of two varieties of Douglas fir (Pseudotsuga menziesii var. viridis and glauca).

Douglas fir (Pseudotsuga menziesii) variety glauca (DFG) but not the variety viridis (DFV) showed symptoms of manganese (Mn) toxicity in some field sites. We hypothesized that these two varieties differed in Mn metabolism. To test this hypothesis, biomass partitioning, Mn concentrations, subcellular localization and 54Mn-transport were investigated. Total Mn uptake was three-times higher in DFG than in DFV. DFV retained > 90% of 54Mn in roots, whereas > 60% was transported to the shoot in DFG. The epidermis was probably the most efficient Mn barrier since DFV contained lower Mn concentrations in cortical cells and vacuoles of roots than DFG. In both varieties, xylem loading was restricted and phloem transport was low. However, sieve cells still contained high Mn concentrations. DFV displayed higher biomass production and higher shoot : root ratios than DFG. Our results clearly show that both varieties of Douglas fir differ significantly in Mn-uptake and allocation patterns rendering DFG more vulnerable to Mn toxicity.

Storage versus substrate limitation to bole respiratory potential in two coniferous tree species of contrasting sapwood width.

Two coniferous tree species of contrasting sapwood width (Pinus ponderosa L., ponderosa pine and Pseudotsuga menziesii Mirb., Douglas-fir) were compared to determine whether bole respiratory potential was correlated with available storage space in ray parenchyma cells and/or respiratory substrate concentration of tissues (total nitrogen content, N; and total non-structural carbohydrate content, TNC). An increment core-based, laboratory method under controlled temperature was used to measure tissue-level respiration (termed respiratory potential) from multiple positions in mature boles (>100-years-old). The most significant tissue-level differences that occurred were that N and TNC were two to six times higher for inner bark than sapwood, TNC was about two times higher in ponderosa pine than Douglas-fir and there was significant seasonal variation in TNC. Ray cell abundance was not correlated with sapwood respiratory potential, whereas N and TNC often were, implying that respiratory potential tended to be more limited by substrate than storage space. When scaled from cores to whole boles (excluding branches), potential net CO2 efflux correlated positively with live bole volume (inner bark plus sapwood), live bole ray volume, N mass, and TNC mass (adjusted R2 > or =0.4). This relationship did not differ between species for N mass, but did for live bole volume, live bole ray volume, and TNC mass. Therefore, N mass appeared to be a good predictor of bole respiratory potential. The differences in net CO2 efflux between the species were largely explained by the species' relative amounts of whole-bole storage space or substrate mass. For example, ponderosa pine's inner bark was thinner than Douglas-fir's, which had the greater concentration of ray cells and TNC compared with the sapwood. This resulted in ponderosa pine boles having 30-60% less ray volume and 10-30% less TNC mass, and caused ponderosa pine net CO2 efflux/ray volume and net CO2 efflux/TNC mass to be 20-50% higher than Douglas-fir. In addition, because inner bark respiratory potential was 2-25 times higher than that of sapwood, ponderosa pine's thinner inner bark and deeper sapwood (relative to Douglas-fir) caused its bole net CO2 efflux/live bole volume to be 20-25% lower than that of similarly-sized Douglas-fir trees.

Changes in anatomy and terpene chemistry in roots of Douglas-fir seedlings following treatment with methyl jasmonate.

Replicated trials were conducted on two full-sibling families of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) seedlings. In response to the application of a 0.01% solution of methyl jasmonate (MeJA) to the soil of potted seedlings, numerous anatomical and chemical changes were observed in the roots, stem and foliage. These changes were, for the most part, similar for both sib groups. Methyl jasmonate induced traumatic resin duct formation in roots and stems. Chemical differences between MeJA-treated and control seedlings were mainly limited to the roots and stem, though some changes also occurred in the foliage. A total of 35 terpenoids were observed in the P. menziesii seedlings. In response to MeJA treatment, several of the 22 detected monoterpenoids (linalool, bornyl acetate, camphene, myrcene, alpha- and beta-pinene, tricyclene and beta-phellandrene) increased significantly in roots and stems, whereas (E)-beta-ocimene decreased significantly in the foliage. Four of the five detected sesquiterpenoids (alpha-humulene, germacrene D, longifolene and (E)-caryophyllene) increased significantly following MeJA application, mainly in the root and stem. Four of the eight detected diterpenoids (abietate, levopimarate, palustrate and sandaracopimarate) increased in response to MeJA treatment, but only in root and stem tissue. This study provides the first description of the effects of MeJA applied to roots through the soil on the anatomy and terpene chemistry of a gymnosperm. This comprehensive inventory of terpenoids in P. menziesii, with and without MeJA treatment, may facilitate identification of terpenoid-related resistance traits. Potential practical applications of MeJA treatment of conifer roots as a pest management strategy are discussed.

Reliance on stored water increases with tree size in three species in the Pacific Northwest.

In tall old forests, limitations to water transport may limit maximum tree height and reduce photosynthesis and carbon sequestration. We evaluated the degree to which tall trees could potentially compensate for hydraulic limitations to water transport by increased use of water stored in xylem. Using sap flux measurements in three tree species of the Pacific Northwest, we showed that reliance on stored water increases with tree size and estimated that use of stored water increases photosynthesis. For Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco), water stored in xylem accounted for 20 to 25% of total daily water use in 60-m trees, whereas stored water comprised 7% of daily water use in 15-m trees. For Oregon white oak (Quercus garryana Dougl. ex Hook.), water stored in xylem accounted for 10 to 23% of total daily water use in 25-m trees, whereas stored water comprised 9 to 13% of daily water use in 10-m trees. For ponderosa pine (Pinus ponderosa Dougl. ex Laws.), water stored in xylem accounted for 4 to 20% of total daily water use in 36-m trees, whereas stored water comprised 2 to 4% of daily water use in 12-m trees. In 60-m Douglas-fir trees, we estimated that use of stored water supported 18% more photosynthesis on a daily basis than would occur if no stored water were used, whereas 15-m Douglas-fir trees gained 10% greater daily photosynthesis from use of stored water. We conclude that water storage plays a significant role in the water and carbon economy of tall trees and old forests.

Needle anatomy changes with increasing tree age in Douglas-fir.

Morphological differences between old-growth trees and saplings of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) may extend to differences in needle anatomy. We used microscopy with image analysis to compare and quantify anatomical parameters in cross sections of previous-year needles of old-growth Douglas-fir trees and saplings at the Wind River Canopy Crane site in Washington and at three sites in the Cascade Mountains of Oregon. We also compared needle anatomy across a chronosequence of 10-, 20-, 40- and 450-year-old Douglas-fir trees from the Wind River site. Anatomy differed significantly between needles of old-growth trees and saplings at all sites, suggesting a developmental change in needle anatomy with increasing tree age. Compared with needles of old-growth trees, needles of saplings were longer and had proportionately smaller vascular cylinders, larger resin canals and few hypodermal cells. Astrosclereids, which sequester lignin in their secondary cell walls and occupy space otherwise filled by photosynthetic cells, were scarce in needles of saplings but abundant in needles of old-growth trees. Needles of old-growth trees had an average of 11% less photosynthetic mesophyll area than needles of saplings. The percentage of non-photosynthetic area in needles increased significantly with increasing tree age from the chronosequence of 10-, 20-, 40- and 450-year-old trees at the Wind River site. This reduction in photosynthetic area may contribute to decreased growth rates in old trees.