Differences in delta13C and diameter growth among remnant Scots pine populations in Scotland

Published data suggest that differences in wood cellulose carbon isotope composition (delta13C) and xylem ring width among natural populations of Scots pine in Scotland (Pinus sylvestris L.) are attributable to the persistence of palaeotypes of various post-glacial migratory origins. We assessed differences in wood cellulose delta13C and ring width among Scottish Scots pine populations grown in a clone bank and in natural stands at various locations in northern and central Scotland. Ring width and wood cellulose delta13C varied significantly among natural stands. Potential water deficit was positively correlated with wood cellulose delta13C and xylem ring width in the natural stands. Neither wood cellulose delta13C nor xylem ring width of clone bank trees correlated with any climate variables at the sites from which the trees originated, indicating little adaptation to climate for these traits. Xylem ring width showed a site x population interaction for the growth sites (i.e., natural stands versus clone bank), but wood cellulose delta13C did not. These results suggest that climate variation in Scotland has not resulted in significant genetic variation in wood cellulose delta13C or xylem ring width in post-glacial populations.     

Shoot and root vulnerability to xylem cavitation in four populations of Douglas-fir seedlings

The objectives of this study were to assess the range of genotypic variation in the vulnerability of the shoot and root xylem of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) seedlings to water-stress-induced cavitation, and to assess the trade-off between vulnerability to cavitation and conductivity per unit of stem cross-sectional area (k(s)), both within a species and within an individual tree. Douglas-fir occupies a broad range of environments and exhibits considerable genetic variation for growth, morphology, and drought hardiness. We chose two populations from each of two varieties (the coastal var. menziesii and the interior var. glauca) to represent environmental extremes of the species. Vulnerability curves were constructed for shoots and roots by plotting the percentage loss in conductivity versus water potential. Vulnerability in shoot and root xylem varied genetically with source climate. Stem xylem differed in vulnerability to cavitation between populations; the most mesic population, coastal wet (CW), was the most susceptible of the four populations. In the roots, the most vulnerable population was again CW; the interior wet (IW) population was moderately susceptible compared with the two dry populations, coastal dry (CD) and interior dry (ID). Root xylem was more susceptible to cavitation than stem xylem and had significantly greater k(s). The trade-off between vulnerability to cavitation and k(s), however, was not evident across populations. The most vulnerable population (CW) had a shoot k(s) of 0.534 +/- 0.067 &mgr;mol m(-2) s(-1) MPa(-1), compared with 0.734 +/- 0.067 &mgr;mol m(-2) s(-1) MPa(-1) for the less vulnerable CD stems. In the roots, IW was more vulnerable than ID, but had the same k(s).     

Variation in foliar delta(13)C values within the crowns of Pinus radiata trees

Although herbaceous species generally show little within plant variation in delta(13)C, trees show large spatial and temporal differences. We found that the aspect of exposure and branch length accounted for up to 6 per thousand delta(13)C difference within the foliage of individual trees of Pinus radiata D. Don. The foliage on branches 0.5 m in length was as much as 4 per thousand more depleted in (13)C than foliage on 10-m long branches, and an additional 2 per thousand more depleted on the shaded side than on the exposed side. We confirmed that on clear days, relative branch hydraulic conductivity, defined as the ratio of transpiration to the water potential gradient, was much higher in short branches than in long branches. Stomatal conductance remained high in foliage on short branches during the day, whereas it declined progressively in long-branch foliage under similar conditions. These differences were sufficient to explain the observed variation in delta(13)C in foliage on long and short branches.     

Responses of foliar delta13C, gas exchange and leaf morphology to reduced hydraulic conductivity in Pinus monticola branches

We tested the hypothesis that branch hydraulic conductivity partly controls foliar stable carbon isotope ratio (delta13C) by its influence on stomatal conductance in Pinus monticola Dougl. Notching and phloem-girdling treatments were applied to reduce branch conductivity over the course of a growing season. Notching and phloem girdling reduced leaf-specific conductivity (LSC) by about 30 and 90%, respectively. The 90% reduction in LSC increased foliar delta13C by about 1 per thousand (P < 0.0001, n = 65), whereas the 30% reduction in LSC had no effect on foliar delta13C (P = 0.90, n = 65). Variation in the delta13C of dark respiration was similar to that of whole-tissues when compared among treatments. These isotopic measurements, in addition to instantaneous gas exchange measurements, suggested only minor adjustments in the ratio of intercellular to atmospheric CO2 partial pressures (ci/ca) in response to experimentally reduced hydraulic conductivity. A strong correlation was observed between stomatal conductance (gs) and photosynthetic demand over a tenfold range in gs. Although ci/ca and delta13C appeared to be relatively homeostatic, current-year leaf area varied linearly as a function of branch hydraulic conductivity (r2 = 0.69, P < 0.0001, n = 18). These results suggest that, for Pinus monticola, adjustment of leaf area is a more important response to reduced branch conductivity than adjustment of ci/ca.     

Physiological variation among Populus fremontii populations: short- and long-term relationships between delta13C and water availability

Different populations of widely distributed species can experience dramatically different climatic conditions that may influence physiological activity, specifically carbon assimilation and water use. Populus fremontii Wats. (Fremont cottonwood) populations are found near rivers of varying size along a precipitation gradient from New Mexico to northern California. Climatic differences among populations may lead to physiological differences because P. fremontii is sensitive to water availability. To assess physiological variation among populations, we collected foliage and wood samples from 13 populations that experience different precipitation and stream flow regimes and analyzed the samples for carbon isotope composition (delta13C). Wood delta13C served as a lifetime-averaged indicator of water-use efficiency (WUE), whereas foliage delta13C provided as an estimate of WUE during the growing season of collection. We found approximately 3.4 per thousand variation in delta13C among populations for both foliage (-31.1 to -27.9 per thousand) and wood (-28.3 to -24.7 per thousand). Wood delta13C was, on average, 2.8 per thousand more enriched than foliage. Some of the variation in wood delta13C can be explained by variation in elevation of the study sites. We constructed total precipitation and mean stream flow variables based on the length of the growing season at each study site and analyzed for a relationship between delta13C, precipitation and stream flow. A significant relationship between foliage delta13C and precipitation was found, but water availability did not explain a significant fraction of the variation in wood delta13C. The data suggest that water availability can account for some of the delta13C variation among populations but, given the large residual variances, other factors are important.     

Soil compaction effects on water status of ponderosa pine assessed through 13C/12C composition

Soil compaction is a side effect of forest reestablishment practices resulting from use of heavy equipment and site preparation. Soil compaction often alters soil properties resulting in changes in plant-available water. The use of pressure chamber methods to assess plant water stress has two drawbacks: (1) the measurements are not integrative; and (2) the method is difficult to apply extensively to establish seasonal soil water status. We evaluated leaf carbon isotopic composition (delta13C) as a means of assessing effects of soil compaction on water status and growth of young ponderosa pine (Pinus ponderosa var. ponderosa Dougl. ex Laws) stands across a range of soil textures. Leaf delta13C in cellulose and whole foliar tissue were highly correlated. Leaf delta13C in both whole tissue and cellulose (holocellulose) was up to 1.0 per thousand lower in trees growing in non-compacted (NC) loam or clay soils than in compacted (SC) loam or clay soils. Soil compaction had the opposite effect on leaf delta13C in trees growing on sandy loam soil, indicating that compaction increased water availability in this soil type. Tree growth response to compaction also varied with soil texture, with no effect, a negative effect and a positive effect as a result of compaction of loam, clay and sandy loam soils, respectively. There was a significant correlation between 13C signature and tree growth along the range of soil textures. Leaf delta13C trends were correlated with midday stem water potentials. We conclude that leaf delta13C can be used to measure retrospective water status and to assess the impact of site preparation on tree growth. The advantage of the leaf delta13C approach is that it provides an integrative assessment of past water status in different aged leaves.     

Laser ablation-combustion-GC-IRMS--a new method for online analysis of intra-annual variation of delta13C in tree rings

We present a new, rapid method for high-resolution online determination of delta13C in tree rings, combining laser ablation (LA), combustion (C), gas chromatography (GC) and isotope ratio mass spectrometry (IRMS) (LA-C-GC-IRMS). Sample material was extracted every 6 min with a UV-laser from a tree core, leaving 40-microm-wide holes. Ablated wood dust was combusted to CO2 at 700 degrees C, separated from other gases on a GC column and injected into an isotope ratio mass spectrometer after removal of water vapor. The measurements were calibrated against an internal and an external standard. The tree core remained intact and could be used for subsequent dendrochronological and dendrochemical analyses. Cores from two Scots pine trees (Pinus sylvestris spp. sibirica Lebed.) from central Siberia were sampled. Inter- and intra-annual patterns of delta13C in whole-wood and lignin-extracted cores were indistinguishable apart from a constant offset, suggesting that lignin extraction is unnecessary for our method. Comparison with the conventional method (microtome slicing, elemental analysis and IRMS) indicated high accuracy of the LA-C-GC-IRMS measurements. Patterns of delta13C along three parallel ablation lines on the same core showed high congruence. A conservative estimate of the precision was +/- 0.24 per thousand. Isotopic patterns of the two Scots pine trees were broadly similar, indicating a signal related to the forest stand's climate history. The maximum variation in delta13C over 22 years was about 5 per thousand, ranging from -27 to -22.3 per thousand. The most obvious pattern was a sharp decline in delta13C during latewood formation and a rapid increase with spring early growth. We conclude that the LA-C-GC-IRMS method will be useful in elucidating short-term climate effects on the delta13C signal in tree rings.     

Roles of soil chemistry and water availability in site-related delta(13)C variations in French beech forests

The carbon isotopic composition (delta(13)C) of wood and leaf cellulose of beech trees (Fagus sylvatica L.) was studied at 80 sites in northeastern France. We sampled sites with contrasting water balance, depending on soil type and precipitation. We tested the hypothesis that inter-site variations in plant delta(13)C reflect the spatial distribution of soil water availability, and we assessed whether delta(13)C could be used as a bioindicator of soil water availability. Patterns of variation in delta(13)C were compared with estimates of monthly water balance and with other soil characteristics. Between-site variability in delta(13)C was high (2.9 per thousand range in wood cellulose, 2.1 per thousand in leaf cellulose), but variation in water availability appeared to be only a minor factor contributing to this variation in delta(13)C. Unexpectedly, spatial variations in wood and leaf cellulose delta(13)C were significantly and positively related to soil fertility expressed by soil pH (r = 0.42 and 0.43, respectively) and cation content. On average, trees growing on acidic soils displayed 0.5 per thousand lower delta(13)C in both wood and leaf material than trees growing on neutral or calcareous soils. Our initial hypothesis of a strong negative relationship between delta(13)C and site water availability was not confirmed. In the study zone, neither wood nor leaf delta(13)C appeared to be a reliable bioindicator of spatial variations in water availability. Possible causes for the lack of a relationship are discussed. Our findings confirm, under natural conditions, the strong effect of soil fertility on water-use efficiency previously observed in experiments. This effect needs to be considered in isotopic studies involving different sites.     

Analyses of delta(13)C and delta(18)O in tree rings of Callitris columellaris provide evidence of a change in stomatal control of photosynthesis in response to regional changes in climate

We examined relationships between stable isotopes of carbon (delta(13)C) and oxygen (delta(18)O) in tree rings of Callitris columellaris F. Muell in the semi-arid Pilbara region of north-western Australia. To test the hypothesis that stomatal control of photosynthesis decreases during drier periods, we developed delta(13)C and delta(18)O chronologies spanning 1919-1999, and used a permutation regression approach to relate a 21-year running correlation between delta(13)C and delta(18)O to rainfall and temperature at Marble Bar and our study site. The relationship between delta(13)C and delta(18)O switched from being always negative before 1955 to being consistently positive after 1976, suggesting an increase in stomatal control of photosynthesis in recent decades. Changes in the delta(13)C-delta(18)O relationship reflected changes in rainfall, which has increased in the region by 30% since 1976. The correlation between delta(13)C and delta(18)O was positively related to the 21-year running mean of normalized rainfall anomalies at both the study site (P = 0.045, Adj. r(2) = 0.47) and Marble Bar (P = 0.046, Adj. r(2) = 0.48). In addition, the delta(13)C-delta(18)O correlation was negatively related (P = 0.047, Adj. r(2) = 0.61) to temperatures at Marble Bar. Our interpretation of the role of changes in climate affecting the relationship between tree-ring delta(13)C and delta(18)O is supported by evidence from the isotope composition of foliage samples: foliar delta(13)C and delta(18)O were negatively correlated with log stomatal conductance (delta(13)C, r = -0.41; delta(18)O, r = -0.42), whereas the correlation between foliar delta(13)C and delta(18)O was positive (r = 0.63, P = 0.027) after the summer wet period. Our data indicate that stomatal control of photosynthesis in Callitris adjusts to region-wide changes in climate and that, in a warmer and drier world, trees might adapt by increasing non-stomatal control of photosynthesis.     

Water availability and branch length determine delta(13)C in foliage of Pinus pinaster

The stable carbon isotope composition (delta(13)C) of foliage integrates signals resulting from environmental and hydraulic constraints on water movement and photosynthesis. We used branch length as a simple predictor of hydraulic constraints to water fluxes and determined the response of delta(13)C to varying water availability. Foliage up to 6 years old was taken from Pinus pinaster Ait. trees growing at four sites differing in precipitation (P; 414-984 mm year(-1)) and potential evaporation (ET; 1091-1750 mm year(-1)). Branch length was the principal determinant of temporal trends in delta(13)C. The strong relationship between delta(13)C and branch length was a function of hydraulic conductance, which was negatively correlated with branch length (r(2) = 0.84). Variation in P and ET among sites was reflected in delta(13)C, which was negatively correlated with P/ET (r(2) = 0.66). However, this analysis was confounded by differences in branch length. If the effects of branch length on delta(13)C were first removed, then the 'residual' delta(13)C was more closely related to P/ET (r(2) = 0.99), highlighting the importance of accounting for variation in hydraulic constraints to water flux between sites and years. For plant species that exhibit considerable phenotypic plasticity in response to changes in environment (e.g., variation in leaf area, branch length and number, or stem form), the environmental effects on delta(13)C in foliage can only be reliably assessed if deconvoluted from hydraulic constraints.     

Decreased needle longevity of fertilized Douglas-fir and grand fir in the northern Rockies

Changes in nutrient availability significantly affect canopy dynamics in conifers. To elucidate these effects, we experimentally fertilized mixed conifer stands at several sites across the northern Rocky Mountains. We measured needle longevity, total branch length and foliated length along the main branch axis, and determined mean retained cohort length on mid-canopy branches of shade-intolerant Douglas-fir (Pseudotsuga menziesii var. glauca (Beissn.) Franco) and shade-tolerant grand fir (Abies grandis Lindl.). Needle longevity ranged from 6 to 8 years in Douglas-fir and from 7 to 8 years in grand fir on unfertilized plots. Fertilization significantly decreased needle longevity by 26 and 27% in Douglas-fir and grand fir, respectively. However, the foliated branch length remained unchanged following fertilization and was similar for both species, indicating a 33% increase in mean branch length per needle cohort in Douglas-fir and a 27% increase in grand fir. These data are consistent with the theory that foliated branch length and needle longevity are a result of the ecological light compensation point (ELCP), which links the inherent physiology of the leaf with the availability of resources in the leaf environment. Mid-canopy ELCP was approximately 74 and 71 cm from the branch terminus in Douglas-fir and grand fir, respectively, regardless of fertilization. We hypothesize that fertilization-enhanced needle production and annual shoot growth resulted in a higher rate of shading of older needles. The shaded needles were unable to maintain a positive carbon balance and abscised. The results demonstrate that foliated branch length of Douglas-fir and grand fir in the northern Rocky Mountains can be treated as a homeostatic response to fertilization, whereas foliar turnover is plastic.     

Co-occurring species differ in tree-ring delta(18)O trends

The stable oxygen isotope ratio (delta(18)O) of tree-ring cellulose is jointly determined by the delta(18)O of xylem water, the delta(18)O of atmospheric water vapor, the humidity of the atmosphere and perhaps by species-specific differences in leaf structure and function. Atmospheric humidity and the delta(18)O of water vapor vary seasonally and annually, but if the canopy atmosphere is well mixed, atmospheric characteristics should be uniform among co-occurring trees. In contrast, xylem water delta(18)O is determined by the delta(18)O of water being drawn from the soil, which varies with depth. If co-occurring trees draw water from different soil depths, this soil-water delta(18)O signal would be manifest as differences in delta(18)O among the trees. We examined the variation in tree ring delta(18)O, over eight decades during the 20th Century, among three species co-occurring in natural forest stands of the northern Rocky Mountains in the USA. We sampled 10 Douglas-firs (Pseudotsuga menziesii (Mirb.) Franco var. glauca), 10 ponderosa pines (Pinus ponderosa Laws.) and seven western white pines (Pinus monticola Dougl.). As expected, variation in atmospheric conditions was recorded in the delta(18)O of the cellulose produced in a given year, but observed climatic correlations with delta(18)O were weak. Significant correlations with June climate data included: daily maximum temperature (r = 0.29), daily minimum temperature (r = -0.25), mean temperature (r = 0.20), mean daily precipitation (r = -0.54), vapor pressure deficit (r = 0.32) and solar radiation (r = 0.44). Lagged effects were observed in Douglas-fir and western white pine. In these species, the delta(18)O of a given annual ring was correlated with the delta(18)O of the previous ring. Ponderosa pine showed no significant autocorrelation. Although the species means were correlated among years (r = 0.67 to 0.76), ponderosa pine was consistently enriched in delta(18)O relative to the other species; differences were close to 2 per thousand and they are steadily increasing. Relative to the mean for the three species, ponderosa pine is becoming steadily more enriched (-1.0 per thousand). In contrast, Douglas-fir is being steadily depleted and western pine is intermediate, with an enrichment of 0.5 per thousand. Because all trees were exposed to the same atmospheric conditions, the differences in delta(18)O observed between species are likely due either to differences in the depth of water extraction or leaf function. If the former, presumably ponderosa pine has steadily taken up more water from near the soil surface and Douglas-fir has shifted uptake to a greater depth. If the latter, we suggest the pronounced changes in leaf-water delta(18)O are a result of changes in leaf structure and function with tree size and age.     

Environmental and physiological controls over oxygen and carbon isotope composition of Tasmanian blue gum, Eucalyptus globulus

We measured oxygen isotope ratios (delta18O) of xylem sap, phloem sap, leaves, wood and bark of Eucalyptus globulus Labill. growing in southwestern Australia. Carbon isotope ratios (delta13C) were measured in the dry matter of phloem sap, leaves and wood. Results were used to test several aspects of a mechanistic model of 18O enrichment and provided insights into post-photosynthetic variations in dry matter delta13C. Xylem water delta18O varied little within the tree crown, whereas variation at the landscape-level was more pronounced, with plantations near the coast being enriched by up to 3 per thousand compared with plantations less than 100 km inland. Phloem water was significantly enriched in 18O compared with xylem water in two of three sampling campaigns; mean enrichments were 0.5 and 0.8 per thousand. Phloem sap sugars exported from E. globulus leaves closely reflected observed leaf water enrichment when diurnal variation in photosynthesis was taken into account. Photosynthetic rates were higher in the morning than in the afternoon, whereas leaf water 18O enrichment increased to maximum values in the afternoon. A non-steady-state model of leaf water 18O enrichment accurately predicted observed values through a full diel cycle. Mean estimates of the proportion of organic oxygen effectively exchanging with xylem water during cellulose synthesis were close to 0.40 for both leaves and wood. Carbon isotope ratios of nascent xylem tissues did not differ from those of phloem sap sugars collected concurrently, whereas nascent leaf tissues were depleted in 13C by 2 per thousand compared with phloem sap sugars, suggesting that, in E. globulus, 13C enrichment of sink tissues compared with source leaves does not result from an enriching process within the sink tissue.     

Correlations between stable carbon-isotope abundance and hydraulic conductivity in Douglas-fir across a climate gradient in Oregon, USA

Stomatal conductance in trees is related to both foliar carbon-isotope abundance and stem hydraulic properties. By combining these relationships, I hypothesized that carbon-isotope abundance in foliage should vary with limitations to water movement through supporting branches. I sampled Douglas-fir branches (Pseudotsuga menziesii (Mirb.) Franco) from six sites across a climate gradient in Oregon, USA for foliar carbon-isotope abundance and stem hydraulic properties. I used a forest growth model to quantify climate-induced stomatal limitations, expressed as reduced potential transpiration, across the gradient. Foliar stable carbon-isotope abundance showed a strong inverse relationship with branch specific conductivity (hydraulic conductivity per unit functional sapwood area) and leaf-specific conductivity (hydraulic conductivity per unit leaf area). Foliar stable carbon-isotope abundance was correlated with modeled reductions in potential transpiration; however, the inclusion of leaf-specific conductivity improved the correlation by more than 30%. Combined, leaf-specific conductivity and climate-induced stomatal constraints explained 84% of the variation in foliar isotope abundance in 1994 foliage. This model was confirmed on foliage classes 1990-1993.     

Foliar nitrogen concentrations and natural abundance of (15)N suggest nitrogen allocation patterns of Douglas-fir and mycorrhizal fungi during development in elevated carbon dioxide concentration and temperature

Pseudotsuga menziesii (Mirb.) Franco (Douglas-fir) seedlings were grown in a 2 x 2 factorial design in enclosed mesocosms at ambient temperature or 3.5 degrees C above ambient, and at ambient CO2 concentration ([CO2]) or 179 ppm above ambient. Two additional mesocosms were maintained as open controls. We measured the extent of mycorrhizal infection, foliar nitrogen (N) concentrations on both a weight basis (%N) and area basis (Narea), and foliar delta15N signatures (15N/14N ratios) from summer 1993 through summer 1997. Mycorrhizal fungi had colonized nearly all root tips across all treatments by spring 1994. Elevated [CO2] lowered foliar %N but did not affect N(area), whereas elevated temperature increased both foliar %N and Narea. Foliar delta15N was initially -1 per thousand and dropped by the final harvest to between -4 and -5 per thousand in the enclosed mesocosms, probably because of transfer of isotopically depleted N from mycorrhizal fungi. Based on the similarity in foliar delta15N among treatments, we conclude that mycorrhizal fungi had similar N allocation patterns across CO2 and temperature treatments. We combined isotopic and Narea data for 1993-94 to calculate fluxes of N for second- and third-year needles. Yearly N influxes were higher in second-year needles than in third-year needles (about 160 and 50% of initial leaf N, respectively), indicating greater sink strength in the younger needles. Influxes of N in second-year needles increased in response to elevated temperature, suggesting increased N supply from soil relative to plant N demands. In the elevated temperature treatments, N effluxes from third-year needles were higher in seedlings in elevated [CO2] than in ambient [CO2], probably because of increased N allocation below ground. We conclude that N allocation patterns shifted in response to the elevated temperature and [CO2] treatments in the seedlings but not in their fungal symbionts.     

Leaf and wood carbon isotope ratios, specific leaf areas and wood growth of Eucalyptus species across a rainfall gradient in Australia

Leaves and samples of recent wood of Eucalyptus species were collected along a rainfall gradient parallel to the coast of Western Australia between Perth in the north and Walpole in the south and along a southwest to northeast transect from Walpole in southwestern Australia, to near Mount Olga in central Australia. The collection included 65 species of Eucalyptus sampled at 73 sites and many of the species were collected at several sites along the rainfall gradient. Specific leaf area (SLA) and isotopic ratio of 13C to 12C (delta 13C) of leaves that grew in 2002, and tree ring growth and delta 13C of individual cell layers of the wood were measured. Rainfall data were obtained from the Australian Bureau of Meteorology for 29 locations that represented one or a few closely located collection sites. Site-averaged data and species-specific values of delta 13C decreased with decreasing annual rainfall between 1200 and 300 mm at a rate of 1.63 per thousand per 1000 mm decrease in rainfall. Responses became variable in the low rainfall region (< 300 mm), with some species showing decreasing delta 13C with rainfall, whereas delta 13C increased or remained constant in other species. The range of delta 13C values in the low rainfall region was as large as the range observed at sites receiving > 300 mm of annual rainfall. Specific leaf area varied between 2 and 6 m2 kg(-1) and tended to increase with decreasing annual rainfall in some species, but not all, whereas delta 13C decreased with SLA. The relationship between delta 13C and SLA was highly species and soil-type specific. Leaf-area-based nitrogen (N) content varied between 2 and almost 6 g m(-2) and decreased with rainfall. Thus, thicker leaves were associated with higher N content and this compensated for the effect of drought on delta 13C. Nitrogen content was also related to soil type and species identity. Based on a linear mixed model, statistical analysis of the whole data set showed that 27% of the variation in delta 13C was associated with changes in SLA, 16% with soil type and only 1% with rainfall. Additionally, 21% was associated with species identity. For a subset of sites with > 300 mm rainfall, 43% of the variation was explained by SLA, 13% by soil type and only 3% by rainfall. The species effect decreased to 9% because there were fewer species in the subset of sites. The small effect of rainfall on delta 13C was further supported by a path analysis that yielded a standardized path coefficient of 0.38 for the effect of rainfall on SLA and -0.50 for the effect of SLA on delta 13C, but an insignificantly low standardized path coefficient of -0.05 for the direct effect of rainfall on delta 13C. Thus, in contrast to our hypothesis that delta 13C decreases with rainfall independent of soil type and species, we detected no statistically significant relationship between rainfall and delta 13C in leaves of trees growing at sites receiving < 300 mm of rainfall annually. Rainfall affected delta 13C indirectly through soil type (a surrogate for water-holding capacity) across the rainfall gradient. Annual tree rings are not clearly visible in evergreen Eucalyptus species, even in the seasonally cool climate of SW Australia. Generally, visible density transitions in the wood are related not to a strict annual cycle but to periods of growth associated mainly with rainfall. The relationship between delta 13C of leaves and the width of these stem increments was not statistically significant. Analysis of stem growth periods showed that delta 13C in wood responded to rainfall events, but carbohydrate storage and reallocation also affected the isotopic signature. Although delta 13C in wood of any one species varied over a range of 2 to 4 per thousand, there was a general relationship between delta 13C of the leaves and the annual range of delta 13C in wood. We conclude that species-specific traits are important in understanding the response of Eucalyptus to rainfall and that the diversity of the genus may reflect its response to the large climatic gradient in Australia and to the large annual and interannual variations in rainfall at any one location.     

A multi-species comparison of delta13C from whole wood, extractive-free wood and holocellulose

The stable carbon (C) isotope composition (delta13C) of tree rings is a powerful metric for reconstructing past physiological responses to climate variation. However, accurate measurement and interpretation are complicated by diagenesis and the translocation of compounds with distinct isotopic signatures. Isolation and analysis of cellulose minimizes these complications by eliminating variation due to biosynthetic pathways; however, isolation of cellulose is time-consuming and has no clear endpoint. A faster and better-defined analytical method is desirable. Our objectives were to determine if there is a direct relationship between the isotopic compositions of whole wood (WW), whole wood treated with solvents to remove mobile extractives (extractive-free wood; EF) and holocellulose (HC) isolated by extractive removal and subsequent bleaching. We also determined if total C concentration could explain the isotopic composition and variation among these three wood components of each sample. A set of wood samples of diverse phylogeny, anatomy and chemical composition, was examined. The mean offset or difference between HC and EF delta13C was 1.07 +/- 0.09 per thousand and the offset between HC and WW was 1.32 +/- 0.10 per thousand. Equivalence tests (with alpha = 0.05) indicated that the relationship between EF delta13C and HC delta13C had a slope significantly similar to 1 +/- 5.5%, whereas for the WW delta13C: HC delta13C relationship, the slope was significantly similar to 1 +/- 10.08%. A regression model using EF delta13C to predict HC delta13C had a slope of 0.97, which was not significantly different from unity (P = 0.264), whereas the regression for WW had a slope of 0.92 which was significantly different from unity (P = 0.0098). Carbon concentration was correlated with HC:WW offset and cellulose:EF offset (P = 0.0501 and 0.007, respectively), but neither relationship explained much of the variation (r2 = 0.12 and 0.14, respectively). We suggest that HC extraction is unnecessary for most analyses of tree-ring delta13C; a simple solvent extraction is a suitable alternative for many applications.     

Long-term trends in cellulose delta13 C and water-use efficiency of tropical Cedrela and Swietenia from Brazil

Elevated CO(2) concentrations ([CO(2)]) affect plant water relations and photosynthesis, and the increase in atmospheric [CO(2)] over the past 100-200 years has been related to changes in stomatal density and the carbon isotope ratio (delta(13)C) in tree rings and leaves from herbarium specimens. Because many tropical trees do not produce annual growth rings and their wood is therefore difficult to date, no trends in delta(13)C of tropical trees have been reported. Wood from Cedrela odorata L. (tropical cedar) and Swietenia macrophylla King (bigleaf mahogany), which do produce annual rings, was collected from a primary rain forest in Aripuan?, Brazil (10 degrees 09' S, 59 degrees 26' W). We measured wood cellulose delta(13)C in 10-year growth increments from 37 Cedrela trees (between 11 and 151 years old in 2001) and 16 Swietenia trees (48-126 years old). A comparison of delta(13)C in cellulose of trees from different decades and of trees of different cambial ages showed that the amount of delta(13)C was largely related to the decade the wood was produced in, and not, or only to a minor extent, to tree age. Cellulose delta(13)C decreased from -26.0 to -27.3 per thousand in Cedrela and from -25.7 to -27.1 per thousand in Swietenia, with the largest changes occurring during the past 50 years. Based on these data and the trends in atmospheric [CO(2)] and delta(13)CO(2), we calculated that the internal [CO(2)] increased from about 220 to 260 ppm and that intrinsic water-use efficiency increased by 34% in Cedrela and by 52% in Swietenia. This may have implications for the water cycle and may explain the trend toward increased tree growth and turnover observed in some tropical forests.     

Hydraulic properties of Douglas-fir (Pseudotsuga menziesii) branches and branch halves with reference to compression wood

Douglas-fir (Pseudotsuga menziesii var. menziesii (Mirb.) Franco) branch segments were used to test the hypothesis that compression wood reduces xylem transport efficiency. Whole 3-year-old segments were first measured for specific conductivity (k(s), m(2) s(-1) MPa(-1)), then split lengthwise into upper and lower halves, the latter containing all or most of the compression wood in the segment. Halves were then remeasured for k(s) using a new technique that prevents leakage of permeating fluid during measurements. Lower branch halves had significantly lower k(s) than upper halves (6.4 +/- 0.3 versus 9.3 +/- 0.3 m(2) s(-1) MPa(-1) x 10(-4), respectively; n = 36), and despite their larger size, significantly lower hydraulic conductivity (k(h), m(4) s(-1) MPa(-1)) than upper halves. Lower branch halves had higher specific gravity (0.51 +/- 0.01 versus 0.45 +/- 0.01; n = 36), lower water content (123 +/- 2% versus 155 +/- 3%; n = 36), and larger proportions of volume occupied by both cell wall and air than upper halves. Lower halves had more tracheids per annual ring than upper halves (73 +/- 3 versus 63 +/- 2 per radial transect, respectively; n = 36), but tracheids were shorter and had narrower lumens than those of upper branch halves. Differences in hydraulic properties between upper and lower halves suggest that compression wood does reduce xylem transport efficiency. In contrast, the amount of compression wood in each sample did not explain any variation in whole unsplit sample hydraulic properties.     

Age-related variations in delta(13)C of ecosystem respiration across a coniferous forest chronosequence in the Pacific Northwest

We tested the hypothesis that forest age influences the carbon isotope ratio (delta13C) of carbon reservoirs and CO2 at local and regional levels. Carbon isotope ratios of ecosystem respiration (delta13C(R)), soil respiration (delta13C(R-soil)), bulk needle tissue (delta13C(P)) and soil organic carbon (delta(13)C(SOC)) were measured in > 450-, 40- and 20-year-old temperate, mixed coniferous forests in southern Washington, USA. Values of delta13C(R), delta13C(R-soil), delta13C(P) and delta13C(SOC) showed consistent enrichment with increasing stand age. Between the youngest and oldest forests there was an approximately 1 per thousand enrichment in delta13C(P) (at similar canopy levels), delta13C(SOC) (throughout the soil column), delta13C(R-soil) (during the wet season) and delta13C(R) (during the dry season). Mean values of delta13C(R) were -25.9, -26.5 and -27.0 per thousand for the 450-, 40- and 20-year-old forests, respectively. Both delta13C(R-soil) and the difference between delta13C(R) and delta13C(R-soil) were more 13C enriched in older forests than in young forest: delta13C(R) - delta13C(R-soil) = 2.3, 1.1 and 0.5 per thousand for the 450-, 40- and 20-year-old forests, respectively. Values of delta(13)C(P) were proportionally more depleted relative to delta13C(R): delta13C(R) - delta13C(P) = 0.5, 2.2 and 2.5 per thousand for the 450-, 40- and 20-year-old forests, respectively. Values of delta13C(P) were most 13C-enriched at the top of the canopy and in the oldest forest regardless of season (overall values were -26.9, -28.7 and -29.4 per thousand for the 450-, 40- and 20-year-old forests, respectively). Values of delta13C(SOC) from shallow soil depths were similar to delta13C(P) values of upper- and mid-canopy needles. All delta13C data are consistent with the hypothesis that a decrease in stomatal conductance associated with decreased hydraulic conductance leads to increased CO2 diffusional limitations in older coniferous trees. The strong associations between delta13C(P) in needles with delta13C(R) and delta13C(R-soil) at the forest level suggest that 13C observations scale between leaf and ecosystem levels.