Internal cycling of nitrogen, potassium and magnesium in young Sitka spruce.

Potassium (K) and magnesium (Mg) are essential macro-nutrients, but little is known about how they are cycled within plants. Stable isotope studies have shown that the internal cycling of nitrogen (N) is independent of current nutrient supply in temperate tree species. This is ecologically significant because it allows trees to produce rapid shoot growth in spring independent of current soil N uptake. We used stable isotopes to quantify N, K and Mg in new shoots of Sitka spruce (Picea sitchensis (Bong.) Carr.) seedlings and to compare the relative contributions from current uptake and internal cycling. Two-year-old Sitka spruce seedlings were labeled with (15)N, (41)K and (26)Mg in an abundant or a limited supply for one growing season. The trees were repotted in the subsequent dormant season to prevent further root uptake of enriched isotopes and provided with an abundant or a limited supply of unlabeled nutrients until they were harvested in early summer of the following year. The supply was switched for half the trees in the second year to create four nutrient regimes. Enrichment of (15)N, (41)K and (26)Mg in current-year growth was attributed to internally cycled N, K and Mg uptake from the previous year. The internal cycling of N, K and Mg in new growth was significantly affected by the first-year nutrient treatments. The second-year nutrient supply affected the growth rates of the trees, but had no effect on the amounts of N, K or Mg contributed from internal cycling. Thus, internal cycling of K and Mg in Sitka spruce are, like that of N, independent of current nutrient supply.

Stable isotopes in ecosystem science: structure, function and dynamics of a subtropical Savanna.

Stable isotopes are often utilized as intrinsic tracers to study the effects of human land uses on the structural and functional characteristics of ecosystems. Here, we illustrate how stable isotopes of H, C, and O have been utilized to document changes in ecosystem structure and function using a case study from a subtropical savanna ecosystem. Specifically, we demonstrate that: (1) delta 13C values of soil organic carbon record a vegetation change in this ecosystem from C4 grassland to C3 woodland during the past 40-120 years, and (2) delta 2H and delta 18O of plant and soil water reveal changes in ecosystem hydrology that accompanied this grassland-to-woodland transition. In the Rio Grande Plains of North America, delta 13C values of plants and soils indicate that areas now dominated by C3 subtropical thorn woodland were once C4 grasslands. delta 13C values of current organic matter inputs from wooded landscape elements in this region are characteristic of C3 plants (-28 to -25/1000), while those of the associated soil organic carbon are higher and range from -20 to -15/1000. Approximately 50-90% of soil carbon beneath the present C3 woodlands is derived from C4 grasses. A strong memory of the C4 grasslands that once dominated this region is retained by delta 13C values of organic carbon associated with fine and coarse clay fractions. When delta 13C values are evaluated in conjunction with 14C measurements of that same soil carbon, it appears that grassland-to-woodland conversion occurred largely within the past 40-120 years, coincident with the intensification of livestock grazing and reductions in fire frequency. These conclusions substantiate those based on demographic characteristics of the dominant tree species, historical aerial photography, and accounts of early settlers and explores. Concurrent changes in soil delta 13C values and organic carbon content over the past 90 years also indicate that wooded landscape elements are behaving as sinks for atmospheric CO2 by sequestering carbon derived from both the previous C4 grassland and the present C3 woody vegetation. Present day woodlands have hydrologic characteristics fundamentally different from those of the original grasslands. Compared to plants in remnant grasslands, tree and shrub species in the woodlands are rooted more deeply and have significantly greater root biomass and density than grasslands. delta 18O and delta 2H values of plant and soil water confirm that grassland species acquire soil water primarily from the upper 0.5 m of the soil profile. In contrast, trees and shrubs utilize soil water from throughout the upper 4 m of the profile. Thus, soil water that formerly may have infiltrated beyond the reach of the grassland roots and contributed to local groundwater recharge or other hydrologic fluxes may now be captured and transpired by the recently formed woodland plant communities. The natural abundances of stable isotopes revealed fundamental information regarding the impacts of human land use activities on the structure and function of this subtropical savanna. Stable isotopes provided direct, spatially explicit evidence for dramatic changes in ecosystem physiognomy and demonstrated some functional consequences for the hydrologic cycle. Furthermore, grassland-to-woodland conversion has been geographically extensive in the worlds' drylands, suggesting that these ecosystem-level changes in vegetation structure, carbon cycling, and hydrology may have implications for regional/global biogeochemistry and climate.

Estimation of energy expenditure in free-living red deer (Cervus elaphus) with the doubly-labelled water method.

Energy expenditure was estimated using the doubly-labelled water (DLW) method in summer in five free-living adult, non-pregnant, non-lactating, red deer (Cervus elaphus) hinds (weight 107.3 (SE 0.9) kg; age 6 (SE 1) years) on lowland pasture under typical farming conditions. Climatic conditions were monitored throughout the experiment. Errors due to 2H losses in CH4 and faeces were calculated from previous estimates of stoichiometries. CH4 production, fractionated water loss, urinary N and O2 consumption were estimated using an iterative approach. The water flux (rH2O) in these animals consuming only fresh grass was 12 (SE 0.5) kg/d, the CO2 production (rCO2) was 1271 (SE 40) litres/d and the mean energy expenditure was 25 (SE 0.8) MJ/d. There were no significant differences in the isotope distribution spaces and flux rates, rH2O, rCO2 or energy expenditure using the multi-point or two-point approaches to calculation. The DLW-derived energy expenditure of 25 MJ/d is approximately 20% higher than the recommended intake of 21 MJ/d for adult hinds kept outdoors (Adam, 1986) and, at 757 kJ/kg0.75 per d, one third higher than the value of 570 kJ/kg0.75 per d for stags penned indoors (Key et al. 1984).

Validation in sheep of the doubly labeled water method for estimating CO2 production.

Carbon dioxide production (rCO2) was estimated in four sheep over a period of 10 days using doubly labeled water (2H and 18O) and was compared with simultaneous respiration chamber measurements of CO2. The excess 2H and 18O measurements were corrected for the empirically determined effects of isotope rebreathing within the confines of the chambers. A weighted monoexponential curve was then fitted to the data from which isotope flux rates and ultimately rCO2 and water turnover (rH2O) estimates were made. The curve fits were weighted assuming a Poisson model. Selection of this weighting policy did not bias the results, and curvature in the data also appeared to have little effect on the rCO2 estimates. Fractionated evaporative water loss expressed as a fraction of rH2O (X) was estimated from water balance and breath water production estimates; the mean X was 0.145 and ranged from 0.108 to 0.183. Corrections for 2H loss in fecal solids reduced the mean rH2O (4,746 g/day) by 35.5 g/day and increased the mean rCO2 (332.3 l/day) by 21.2 l/day. Further corrections to account for 2H loss in methane (mean production rate 27.2 l/day) reduced rH2O by 33.8 g/day and increased rCO2 by 20.3 l/day. The final isotopic estimates of rH2O were 14.6 +/- 6.59% (n = 4) lower than direct measurements and the mean rCO2 was 3.5 +/- 14.48% (n = 4) lower than the chamber measured rCO2. However, in one of the animals studied the rCO2 deviated markedly from the chamber-derived value, and this discrepancy has yet to be explained. When this animal was excluded from the comparisons, the standard deviation was greatly reduced (+/- 3.6, n = 3) and the mean overall error on rCO2 was +3.6%.

The doubly labeled water method: errors due to deuterium exchange and sequestration in ruminants.

The doubly labeled water (DLW) technique allows the CO2 production (rCO2) of free living animals to be estimated from the difference between the turnover of 2H2O and H218O in the body water. A fundamental assumption of this technique is that neither of the isotopes used are lost in products other than CO2 and H2O. We found, however, that 2H was lost in both exchangeable and nonexchangeable positions in the feces of sheep. Negligible amounts of 18O were lost in exchangeable positions. 2H losses led to a 0.75% (SE 0.06, n = 4) overestimation of the measured 2H2O flux, leading to an average error in rCO2 estimates of 20.3 l/day. For a typical rCO2 rate of 370 l/day, this would amount to an error of approximately 5% (range -7.0 to -4.3%, n = 4). Correction factors to account for this loss were presented. The error in rCO2 due to 2H sequestration into fat was calculated to be at most 2.1 l/day or about -0.66% in lambs with a rCO2 of 320 l/day. In a triply labeled water (TLW) study the maximum error in the estimation of fractionated evaporative water loss (X) would lead to a 0.81% underestimation of rCO2. We recommend that during a DLW study involving ruminant animals the correction factors presented here be used to compensate for 2H loss in feces. This may be particularly important where the diet has a high roughage content leading to a significant fecal dry matter production.

Stable carbon isotope analysis of soil organic matter illustrates vegetation change at the grassland/woodland boundary in southeastern Arizona, USA.

In southeastern Arizona, Prosopis juliflora (Swartz) DC. and Quercus emoryi Torr. are the dominant woody species at grassland/woodland boundaries. The stability of the grassland/woodland boundary in this region has been questioned, although there is no direct evidence to confirm that woodland is encroaching into grassland or vice versa. We used stable carbon isotope analysis of soil organic matter to investigate the direction and magnitude of vegetation change along this ecotone. δ13C values of soil organic matter and roots along the ecotone indicated that both dominant woody species (C3) are recent components of former grasslands (C4), consistent with other reports of recent increases in woody plant abundance in grasslands and savannas throughout the world. Data on root biomass and soil organic matter suggest that this increase in woody plant abundance in grasslands and savannas may increase carbon storage in these ecosystems, with implications for the global carbon cycle.

Methane production in ruminants: its effect on the doubly labeled water method.

The doubly labeled water (DLW) technique for measuring CO2 production (rCO2) in free-living animals requires an assessment of the elimination of both 2H and 18O from the body over a long period of time. To calculate rCO2, it is necessary to calculate water turnover (rH2O) from the 2H flux rate. In ruminant animals, the accuracy of this calculation is affected by the loss of 2H in methane. We have quantified the effect of methane production (rCH4) on the 2H flux rate, determined in four sheep given 2H2O. The methane produced was depleted in 2H relative to the urine. A relationship between the enrichment of the methane and urine was established. The ratio of urine to methane enrichment was found on average to be 0.6536, and this value was unaffected by the level of rCH4 but showed some dependence on the absolute concentration of 2H in urine. For this reason, the ratio value obtained from four sheep not given 2H2O was different, a mean of 0.6886 was measured, this ratio was unaffected by changes in the diet supplied to the animals. Computer modeling was used to illustrate the dependence of the isotopically derived value for rCO2 on not only rCH4 but also the magnitude of rCO2 itself. The effect of rCH4 on the DLW method can be predicted from the observed ratio of rCO2 to rCH4 and the value of 0.6536 obtained for the ratio of methane to urine enrichment. With the use of data from several studies at this Institute, a limited range of 10 to 20 was found for rCO2/rCH4 in animals fed at or above maintenance.(ABSTRACT TRUNCATED AT 250 WORDS)