Oxygen Isotope Ratios Of Cellulose Research Articles

A predictive spatial model for roasted coffee using oxygen isotopes of α-cellulose.

Fraudulent region-of-origin labeling is a concern for high-value, globally traded commodities such as coffee. The oxygen isotope ratio of cellulose is a useful geographic tracer, as it integrates climate and source water signals. A predictive spatial model ("isoscape") of the δ18 O values of coffee bean cellulose is generated to evaluate coffee region-of-origin claims. The oxygen isotope ratio of α-cellulose extracted from roasted coffee beans was measured via high-temperature conversion elemental analyzer/isotope ratio mass spectrometry (TC-EA/IRMS) and used to calculate the δ18 O value of coffee bean water. The 18 O enrichment of coffee bean water relative to the δ18 O value of local precipitation was modeled as a function of local temperature and humidity. This function was incorporated into a mechanistic model of cellulose δ18 O values to predict the δ18 O values of coffee bean cellulose across coffee-producing regions globally. The δ18 O values of analyzed coffee bean cellulose ranged from approximately +22‰ to +42‰ (V-SMOW). As expected, coffees grown in the same region tended to have similar isotope ratios, and the δ18 O value of coffee bean cellulose was generally higher than the δ18 O value of modeled stem cellulose for the region. Modeled δ18 O values of coffee cellulose were within ±2.3‰ of the measured δ18 O value of coffee cellulose. The oxygen isotope ratio of coffee bean cellulose is a useful indicator of region-of-origin and varies predictably in response to climatic factors and precipitation isotope ratios. The isoscape of coffee bean cellulose δ18 O values from this study provides a quantitative tool that can be applied to region-of-origin verification of roasted coffee at the point-of-sale.

Two Decades of Experimental Manipulation Reveal Potential for Enhanced Biomass Accumulation and Water Use Efficiency in Ponderosa Pine Plantations Across Climate Gradients

AbstractIn this study, we report 20 years of data from three ponderosa pine plantations in northern California. Our sites span a natural gradient of forest productivity where climate variability and edaphic conditions delineate marked differences in baseline productivity (approximately threefold). Experimental herbicide application and fertilization significantly reduced competition and improved tree growth by 1.4‐ to 2.2‐fold across sites. At the site of lowest productivity, where soils are poorly developed and water limiting, tree growth increased strongly in response to understory suppression. Small but significant improvements in tree growth were observed in response to understory suppression at the moderate‐productivity site. At the site of highest productivity, where climate is favorable and soils well developed, fertilization increased productivity to a greater extent than did understory suppression. In most cases, the effect of understory suppression and fertilization caused an unexpected growth release, exceeding the anticipated maximum productivity by >5 m of additional height and 60–100% more basal area. At the site of highest productivity, however, understory suppression caused a weak increase on late‐season growth compared to fertilization alone, suggesting a beneficial effect of understory vegetation on long‐term growth at that site. Tree ring cellulose carbon isotopes indicate a negative relationship between intrinsic water use efficiency (iWUE) and tree growth in control stands, which shifted to a positive relationship as both iWUE and tree growth increased in response to management. Cellulose oxygen isotope ratios (δ18O) were positively correlated with iWUE and negatively correlated with vapor pressure deficit across sites, but δ18O was not a strong predictor of tree growth.

Age determination on a catastrophic rock avalanche using tree-ring oxygen isotope ratios - the scar of a historical gigantic earthquake in the Southern Alps, central Japan

Abstract Dendrochronological analysis using the oxygen isotope composition (18O/16O) of cellulose was performed to reveal the age of an ancient rock avalanche that scattered huge granitic rock clasts on the bank of the Dondokosawa River in the east of Mount Hououzan, the Akaishi Range, central Japan. The samples for dating were disc-cut wood logs or increment cores of Japanese cypress (Chamaecyparis obtusa (Sieb. & Zucc.) Endl.) and Hemlock tree (Tsuga sp.) buried in lacustrine sediments. Cellulose was extracted directly from tree-ring cross-sectional thin wood plates. Cellulose rings of each disc were sampled and their oxygen isotope ratios were measured with a pyrolysis-type elemental analyser and an isotope ratio mass spectrometer. We compared the inter-annual variations in oxygen isotope ratios of cellulose for samples with those of a 470-year-long tree-ring δ18O chronology (AD450–919) independently constructed from Japanese cypress. The estimated death year of the barkless Japanese cypress is around AD885, and that of the barked hemlock tree is AD888. If the rock avalanche was induced by strong ground motion from a large earthquake, these ages are relevant to a large historical earthquake that occurred in this area. The candidates are the AD 887 Goki-Shichido Earthquake (Ninna Earthquake), which was one of the gigantic ocean-trench earthquakes documented along the Suruga and Nankai Troughs off central Japan, and another large intraplate earthquake remotely triggered by this ocean-trench earthquake.

Position-specific measurement of oxygen isotope ratios in cellulose: Isotopic exchange during heterotrophic cellulose synthesis

We describe the first reported method for the measurement of oxygen isotope ratios at each position in the glucose units of the cellulose molecule. The overall process comprises a series of synthetic organic sequences, by which α-cellulose is hydrolysed to glucose, and oxygen atoms at specific positions in the glucose molecule are removed in samples of benzoic acid for measurement of δ18O. Values of δ18O at specific positions in cellulose are calculated from these δ18O values and the overall δ18O value of the cellulose. We apply the method to determine the degree to which oxygen atoms at each position undergo isotopic exchange with water during heterotrophic cellulose synthesis, such as occurs in the cambium of trees. To do this we extract α-cellulose from wheat seedlings germinated in the dark in aqueous media of differing oxygen isotope ratios. Results indicate that oxygen atoms at positions 5 and 6 (O-5 and O-6 respectively) undergo around 80% exchange with medium water, O-3 undergoes around 50% exchange, and O-2 and O-4 do not undergo isotopic exchange. The results have important implications for extracting palaeoclimatic records from oxygen isotope time series obtained from tree ring cellulose. As O-5 and O-6 undergo significant exchange with medium water during heterotrophic cellulose synthesis, oxygen isotopes at these positions in tree ring cellulose should carry a predominantly trunk (source) water signal. On the other hand, O-2 and O-4 should retain the isotopic signature of leaf water in tree ring cellulose. Our method therefore potentially enables the separate reconstruction of past temperature and humidity data from oxygen isotope ratios of tree ring cellulose – something that has hitherto not been possible. The measured degrees of isotopic exchange are to some extent unexpected and cannot be fully explained using current biochemical mechanisms, suggesting that knowledge of these processes is incomplete.

Comparison of stable carbon and oxygen isotopes in Picea glauca tree rings and Sphagnum fuscum moss remains from subarctic Canada

Stable isotope ratios from tree rings and peatland mosses have become important proxies of past climate variations. We here compare recent stable carbon and oxygen isotope ratios in cellulose of tree rings from white spruce (Picea glauca), growing near the arctic tree line; and cellulose of Sphagnum fuscum stems, growing in a hummock of a subarctic peatland, in west-central Canada. Results show that carbon isotopes in S. fuscum correlate significantly with July temperatures over the past ~20yr. The oxygen isotopes correlate with both summer temperature and precipitation. Analyses of the tree-ring isotopes revealed summer temperatures to be the main controlling factor for carbon isotope variations, whereas tree-ring oxygen isotope ratios are controlled by a combination of spring temperatures and precipitation totals. We also explore the potential of combining high-frequency (annual) climate signals derived from long tree-ring series with low-frequency (decadal to centennial) climate signals derived from the moss remains in peat deposits. This cross-archive comparison revealed no association between the oxygen isotopes, which likely results from the varying sensitivity of the archives to different seasons. For the carbon isotopes, common variance could be achieved through adjustments of the Sphagnum age model within dating error.

High‐temperature pyrolysis/gas chromatography/isotope ratio mass spectrometry: simultaneous measurement of the stable isotopes of oxygen and carbon in cellulose

Stable isotope analysis of cellulose is an increasingly important aspect of ecological and palaeoenvironmental research. Since these techniques are very costly, any methodological development which can provide simultaneous measurement of stable carbon and oxygen isotope ratios in cellulose deserves further exploration. A large number (3074) of tree-ring α-cellulose samples are used to compare the stable carbon isotope ratios (δ(13)C) produced by high-temperature (1400°C) pyrolysis/gas chromatography (GC)/isotope ratio mass spectrometry (IRMS) with those produced by combustion GC/IRMS. Although the two data sets are very strongly correlated, the pyrolysis results display reduced variance and are strongly biased towards the mean. The low carbon isotope ratios of tree-ring cellulose during the last century, reflecting anthropogenic disturbance of atmospheric carbon dioxide, are thus overestimated. The likely explanation is that a proportion of the oxygen atoms are bonding with residual carbon in the reaction chamber to form carbon monoxide. The 'pyrolysis adjustment', proposed here, is based on combusting a stratified sub-sample of the pyrolysis results, across the full range of carbon isotope ratios, and using the paired results to define a regression equation that can be used to adjust all the pyrolysis measurements. In this study, subsamples of 30 combustion measurements produced adjusted chronologies statistically indistinguishable from those produced by combusting every sample. This methodology allows simultaneous measurement of the stable isotopes of carbon and oxygen using high-temperature pyrolysis, reducing the amount of sample required and the analytical costs of measuring them separately.

Comparison of whole wood and cellulose carbon and oxygen isotope series from Pinus nigra ssp. laricio (Corsica/France)

Stable isotopes in tree rings have widely been used for palaeoclimate reconstructions since tree rings record climatic information at annual resolution. However, various wood components or different parts of an annual tree-ring may differ in their isotopic compositions. Thus, sample preparation and subsequent laboratory analysis are crucial for the isotopic signal retained in the final tree-ring isotope series used for climate reconstruction and must therefore be considered for the interpretation of isotope–climate relationships. This study focuses on wood of Corsican Pine trees (Pinus nigra ssp. laricio) as this tree species allows to reconstruct the long-term climate evolution in the western Mediterranean. In a pilot study, we concentrated on methodological issues of sample preparation techniques in order to evaluate isotope records measured on pooled whole tree-ring cellulose and whole tree-ring bulk wood samples. We analysed 80-year long carbon and oxygen chronologies of Corsican Pine trees growing near the upper tree line on Corsica. Carbon and oxygen isotope records of whole tree-ring bulk wood and whole tree-ring cellulose from a pooled sample of 5 trees were correlated with the climate parameters monthly precipitation, temperature and the self-calibrating Palmer Drought Severity Index (sc-PDSI). Results show that the offsets in carbon and oxygen isotopes of bulk wood and cellulose are not constant over time. Both isotopes correlate with climate parameters from late winter and summer. The carbon and oxygen isotope ratios of cellulose are more sensitive to climatic variables than those of bulk wood. The results of this study imply that extraction of cellulose is a pre-requisite for the reconstruction of high-resolution climate records from stable isotope series of P. nigra ssp. laricio.

Oxygen stable isotope ratios of tree‐ring cellulose: the next phase of understanding

Analysis of the oxygen isotope ratio of tree-ring cellulose is a valuable tool that can be used as a paleoclimate proxy. Our ability to use this tool has gone through different phases. The first began in the 1970s with the demonstration of empirical relationships between the oxygen isotope ratio of tree-ring cellulose and climate. These empirical relationships, however, did not provide us with the confidence that they are robust through time, across taxa and across geographical locations. The second phase began with a rudimentary understanding of the physiological and biochemical mechanisms responsible for the oxygen isotope ratios of cellulose, which is necessary to increase the power of this tool. This phase culminated in a mechanistic tree-ring model integrating concepts of physiology and biochemistry in a whole-plant system. This model made several assumptions about leaf water isotopic enrichment and biochemistry which, in the nascent third phase, are now being challenged, with surprising results. These third-phase results suggest that, contrary to the model assumption, leaf temperature across a large latitudinal gradient is remarkably constant and does not follow ambient temperature. Recent findings also indicate that the biochemistry responsible for the incorporation of the cellulose oxygen isotopic signature is not as simple as has been assumed. Interestingly, the results of these challenges have strengthened the tree-ring model. There are several other assumptions that can be investigated which will improve the utility of the tree-ring model.

Expressing leaf water and cellulose oxygen isotope ratios as enrichment above source water reveals evidence of a Péclet effect.

There is an increasing ecological interest in understanding the gradients in H(2)(18)O enrichment in leaf water (i.e. a Péclet effect), because an appreciation of the significance of the Péclet effect is important for improving our understanding of the mechanistic processes affecting the (18)O composition of leaf water and plant organic material. In data sets where both source water and leaf water (18)O data are available, we can evaluate the potential contribution of a Péclet effect. As an example, we recalculate data published earlier by Roden and Ehleringer (1999, Oecologia 121:467-477) as enrichments in leaf water (Delta(L)) and cellulose (Delta(cell)) above source water. Based on these recalculations, we present support for the relevance of a Péclet effect in leaves. Further, we demonstrate that the subtle variations in Delta(L) and Delta(cell) caused by a Péclet effect may be masked in experimental systems in which variation in the source water oxygen isotope ratio is considerable.

Grass blades as tree rings: environmentally induced changes in the oxygen isotope ratio of cellulose along the length of grass blades.

• In this study, we tested the hypothesis that environmentally induced changes in the oxygen isotope ratio of leaf water are recorded in grass blade cellulose during leaf-blade expansion. • Grasses were grown hydroponically in chambers that allowed for control of relative humidity while keeping isotopic inputs (namely source water) constant. • In experiments where relative humidity was changed from 35% to 93% during grass blade expansion, a 10‰ shift in cellulose δ 18 O was observed along single grass blades of Lolium multiflorum . However, statistically significant changes were not detectable with relative humidity of 93% to 70%. • It is shown that grass blades, analogously to tree rings, record environmental change on an interseasonal basis. In light of this, care must be taken to compare leaves of the same developmental stage to avoid confusion of environmental effects with physiological effects in interpretations of leaf organic material δ 18 O. The results presented here confirm the ability of the current empirical models to predict the oxygen isotope ratio of cellulose in both grass blades that expanded in constant and variable growth conditions.

Oxygen isotope ratio of leaf and grain material correlates with stomatal conductance and grain yield in irrigated wheat

Theory (Craig and Gordon 1965; Dongmann et al. 1974; Sternberg et al. 1986; Farquhar and Lloyd 1993) suggests that the oxygen isotope ratio (d 18 O) of plant material should reflect the evaporative conditions under which the material was formed, so that differences in stomatal conductance should show up in plant d 18 O. To test this theory we measured the oxygen isotope ratio of organic matter from flag leaves at anthesis and grain at harvest from eight cultivars of spring wheat (Triticum aestivum L.) grown under irrigation in each of three seasons in Mexico. The cultivars ranged widely in stomatal conductance and in average grain yield, with which conductance was positively correlated. Supporting theory, the oxygen isotope ratio of flag leaves (d 18 Ol) was found to correlate negatively with stomatal conductance for two of the three seasons. The significant correlations are consistent with high conductance cultivars having lower leaf temperatures and kinetic fractionation factors, and higher vapour pressure fractionation factors and Péclet numbers, all of which combine to result in less enriched d 18 Ol. Yield (grain weight per square meter) was also found to be significantly negatively correlated with d 18Ol in two of the three seasons. d 18 Ol was as good a predictor of yield as stomatal conductance, and significantly better than carbon isotope discrimination. Correlations between grain d 18 O (d 18 Og) and physiological parameters were less clear. Significant negative correlations between d 18 Og and stomatal conductance, leaf temperature and yield were found only during the first season. By measuring the oxygen isotope ratio of cellulose extracted from leaf samples, the difference in fractionation factors (ecp) for cellulose and whole leaf tissue was assessed. ecp was found to be variable, and more negative when d 18 Oc and d 18 Ol were lower. Cultivar means for d 13 C and d 18 O of whole leaf material were found to be significantly positively related, and the factors required to produce such a relationship are discussed.

Climatic implications of δ13C and δ18O ratios from C3 and C4 plants growing in a tropical montane habitat in southern India

The stable carbon and oxygen isotope ratios in cellulose of C3 and C4 plants growing on the surface of a montane peat bog in the Nilgiri hills, southern India, were measured. The mean monthly δ13C values in cellulose of both C3 and C4 plants are found to be significantly related to rainfall, while the δ18O values are sensitive to changes in maximum temperature and relative humidity of the region. Further, higher δ18O values were observed in C4 plants compared to C3 plants, suggesting that C4 plants are probably less sensitive to relative humidity as compared to C3 plants and are able to photosynthesize even during drier conditions. The plant isotope-climate correlations thus established can be used for reconstructing the past temperature and rainfall conditions of the tropics from the isotopic ratios of peat deposits, derived from a mixture of C3 and C4 plants in the region.

Variations in the natural abundance of oxygen‐18 and deuterium in plant carbohydrates

ABSTRACTVariations in the natural abundance of 18O and 2H in plant cellulose are influenced by the isotopic composition of the water directly involved in metabolism—the metabolic water fraction. The isotopic distinction between the metabolic source water and total tissue water must reflect the formation of isotopic gradients within the tissue that are influenced by the rate of water turnover, by properties of the water conducting system and by environmental conditions. It seems that the 18O abundance in the metabolic water is conserved in cellulose with a relatively constant isotope effect. The relationship of the 2H abundance between metabolic water and cellulose is more complex. Hydrogen incorporated into photosynthetic products during primary reduction steps is highly depleted in 2H. However, a large proportion of these hydrogens are subsequently replaced by exchange with water, leading to 2H enrichment during heterotrophic metabolism. Deciphering the oxygen isotope ratio of cellulose could help in providing insights into the carbon and oxygen fluxes exchanged between plants and the atmosphere. This is because the 18O abundance in cellulose records the 18O abundance in the metabolic water, which in turn, controls the oxygen isotopic signatures of the CO2 and O2 released by plants into the atmosphere. The hydrogen isotope effects associated with carbohydrate metabolism provide insights into the autotrophic state of a plant tissue. This is because the hydrogen isotope ratio of carbohydrates must reflect the net effects of the two opposing isotope effects associated with photosynthesis and heterotrophic metabolism.

Isotopic fractionation during cellulose synthesis in two mangrove species: Salinity effects

Carbon, non-exchangeable hydrogen, and oxygen isotope ratios of cellulose of Avicennia germinans and Rhizophora mangle plants hydroponically grown under different salinities (0, 18, 45% sea water, but with irrigation waters having the same isotopic ratios) were measured to determine the possibility of using isotopic ratios of plant tissues as biological recorders of sea level rise. There was a large variability in the δD values of leaf nitrated cellulose between different treatments and even within a single treatment for both A. germinans and R. mangle. Thus, δD values of non-exchangeable hydrogens of cellulose cannot be used as a historical tracer for utilization of ocean water or freshwater by mangroves. In contrast, δ 18O values of cellulose were not significantly different between different salinity treatments for both mangroves, indicating that δ 18O of cellulose can be used as a sea water tracer. δ 13C values of cellulose did not vary directly with salinity as has been observed with other plants. δ 13C values of cellulose from A. germinans were the lowest for plants growing at 18% sea water, with cellulose from plants growing in 0 and 45% sea water having significantly higher δ 13C values. δ 13C values of cellulose from R. mangle were the highest for plants grown in 45% sea water, with plants grown in 0 and 18% sea water having equally lower δ 13C values.

D/H ratios of environmental water recorded by D/H ratios of plant lipids

Isotope ratios of chemical components are powerful tools in the interpretation of palaeoenvironments, particularly carbonates from foraminiferans1,2, desert caliche3, and desert pavements4. Isotope ratios of plant cellulose are also indicators of environmental variables such as temperature and relative humidity5–7. Several workers have reported climatic fluctuations based on hydrogen and oxygen isotope ratios of cellulose from tree trunks8–10 and peats11,12. Here I present measurements that demonstrate that for submerged aquatic plants δD values of lipids record D/H ratios of environmental water, whereas cellulose does not. I further demonstrate that δD values of lipids in conjunction with δ18O values of cellulose provide significant information on isotope ratios of environmental water.

On the development of a novel method for the determination of stable oxygen isotope ratios in cellulose

We describe a new method for the determination of the 18O/ 16O ratio in tree ring cellulose, that differs greatly from recently published methods which involve the pyrolysis of cellulose or whole wood. By a series of specific chemical reactions, selected oxygen atoms from the cellulose molecule are incorporated into benzoic acid, which upon decarboxylation yields carbon dioxide for mass spectrometric analysis.

Isotopic analysis of archaeobotanicals to reconstruct past climates: Effects of activities associated with food preparation on carbon, hydrogen and oxygen isotope ratios of plant cellulose

The cellulose fraction of botanicals recovered from archaeologically defined strata may be suitable for stable isotope-based palaeoclimatology in much the same manner as cellulose from tree rings has been utilized. However, manipulation by humans is a distinct possibility with such specimens and constitutes a potential source of isotopic alteration early in the post-mortem history of the samples. We studied the susceptibility of plant cellulose to isotopic alteration caused by manipulations related to food processing to determine if discarded plant foods recovered from archaeological contexts are suitable substrates for isotopic palaeoclimatic studies. The carbon and hydrogen isotope ratios of cellulose nitrate and the oxygen isotope ratios of cellulose were determined for Zea mays cobs, Helianthus annus seeds, Agave americana leaves, Pachyrrhizus erosus tubers and a cellulose standard before and after a variety of treatments, including boiling, roasting, fermentation, liming, molding and carbonization. The oxygen and hydrogen isotope ratios (which are climatically sensitive in all plants) and the carbon isotope ratios (which reflect climate only in certain plants) of all samples, except for Agave, were conserved during various food-processing steps. Thus, it appears that the isotope ratios of cellulose and cellulose nitrate prepared from many archaeological food plants, whether processed or uncooked, will reflect in vivo values and can be used to reconstruct aspects of palaeoclimate. Such isotopic palaeoclimatic studies will extend the utility and value of existing collections of ecofacts by opening a unique window for the study of the relationships among climate, plants and man.

Compensation Point and Isotopic Characteristics of C3/C4 Intermediates and Hybrids in Panicum

Leaf CO(2) compensation points and stable hydrogen, oxygen and carbon isotope ratios were determined for Panicum species including C(3)/C(4) intermediate photosynthesis plants, hybrids between C(3)/C(4) intermediates and C(3) plants, C(3) and C(4) plants in the Panicum genus as well as several other C(3) and C(4) plants. C(3) plants had the highest compensation points, followed by hybrids, C(3)/C(4) intermediates, and C(4) plants. delta(13)C values of cellulose nitrate and saponifiable lipids from C(4) plants were about 10 per thousand higher than those observed for cellulose nitrate and saponifiable lipids of C(3)/C(4) intermediates, hybrids, and C(3) plants. Oxygen isotope ratios of cellulose as well as those of leaf water were similar for all plants. There was substantial variability in the deltaD values of cellulose nitrate among the plants studied. In contrast, such variability was not observed in deltaD values of water distilled from the leaves, nor in the deltaD values of the saponifiable lipids. Variability in deltaD values of cellulose nitrate from C(3)/C(4) intermediates, hybrids, C(3), and C(4) plants is due to fractionations occurring during biochemical reactions specific to leaf carbohydrate metabolism.

Isotope Ratios of Cellulose from Plants Having Different Photosynthetic Pathways

Hydrogen and carbon isotope ratios of cellulose nitrate and oxygen isotope ratios of cellulose from C(3), C(4), and Crassulacean acid metabolism (CAM) plants were determined for plants growing within a small area in Val Verde County, Texas. Plants having CAM had distinctly higher deuterium/hydrogen (D/H) ratios than plants having C(3) and C(4) metabolism. When hydrogen isotope ratios are plotted against carbon isotope ratios, each photosynthetic mode separates into a distinct cluster of points. C(4) plants had many D/H ratios similar to those of C(3) plants, so that hydrogen isotope ratios cannot be used to distinguish between these two photosynthetic modes. Portulaca mundula, which may have a modified photosynthetic mode between C(4) and CAM, had a hydrogen isotope ratio between those of the C(4) and CAM plants. When oxygen isotope ratios are plotted against carbon isotope ratios, no distinct clustering of the C(4) and CAM plants occurs. Thus, oxygen isotope ratios are not useful in distinguishing between these metabolic modes. A plot of hydrogen isotope ratios versus oxygen isotope ratios for this sample set shows considerable overlap between oxygen isotope ratios of the different photosynthetic modes without a concomitant overlap in the hydrogen isotope ratios of CAM and the other two photosynthetic modes. This observation is consistent with the hypothesis that higher D/H ratios in CAM plants relative to C(3) and C(4) plants are due to isotopic fractionations occurring during biochemical reactions.

Carbon, Hydrogen, and Oxygen Isotope Ratios of Cellulose from Plants Having Intermediary Photosynthetic Modes

Carbon and hydrogen isotope ratios of cellulose nitrate and oxygen isotope ratios of cellulose from species of greenhouse plants having different photosynthetic modes were determined. When hydrogen isotope ratios are plotted against carbon isotope ratios, four clusters of points are discernible, each representing different photosynthetic modes: C(3) plants, C(4) plants, CAM plants, and C(3) plants that can shift to CAM or show the phenomenon referred to as CAM-cycling. The combination of oxygen and carbon isotope ratios does not distinguish among the different photosynthetic modes. Analysis of the carbon and hydrogen isotope ratios of cellulose nitrate should prove useful for screening different photosynthetic modes in field specimens that grew near one another. This method will be particularly useful for detection of plants which show CAM-cycling.