Pulse-labelling of trees with stable or radioactive carbon (C) isotopes offers the unique opportunity to trace the fate of labelled CO₂ into the tree and its release to the soil and the atmosphere. Thus, pulse-labelling enables the quantification of C partitioning in forests and the assessment of the role of partitioning in tree growth, resource acquisition and C sequestration. However, this is associated with challenges as regards the choice of a tracer, the methods of tracing labelled C in tree and soil compartments and the quantitative analysis of C dynamics. Based on data from 47 studies, the rate of transfer differs between broadleaved and coniferous species and decreases as temperature and soil water content decrease. Labelled C is rapidly transferred belowground—within a few days or less—and this transfer is slowed down by drought. Half-lives of labelled C in phloem sap (transfer pool) and in mature leaves (source organs) are short, while those of sink organs (growing tissues, seasonal storage) are longer. ¹³C measurements in respiratory efflux at high temporal resolution provide the best estimate of the mean residence times of C in respiratory substrate pools, and the best basis for compartmental modelling. Seasonal C dynamics and allocation patterns indicate that sink strength variations are important drivers for C fluxes. We propose a conceptual model for temperate and boreal trees, which considers the use of recently assimilated C versus stored C. We recommend best practices for designing and analysing pulse-labelling experiments, and identify several topics which we consider of prime importance for future research on C allocation in trees: (i) whole-tree C source–sink relations, (ii) C allocation to secondary metabolism, (iii) responses to environmental change, (iv) effects of seasonality versus phenology in and across biomes, and (v) carbon–nitrogen interactions. Substantial progress is expected from emerging technologies, but the largest challenge remains to carry out in situ whole-tree labelling experiments on mature trees to improve our understanding of the environmental and physiological controls on C allocation.

The CO₂ respired by leaves is ¹³C‐enriched relative to leaf biomass and putative respiratory substrates (Ghashghaie et al., Phytochemistry Reviews 2, 145–161, 2003), but how this relates to the ¹³C content of root, or whole plant respiratory CO₂ is unknown. The C isotope composition of respiratory CO₂ (δ_R) from shoots and roots of sunflower (Helianthus annuus L.), alfalfa (Medicago sativa L.), and perennial ryegrass (Lolium perenne L.) growing in a range of conditions was analysed. In all instances plants were grown in controlled environments with CO₂ of constant concentration and δ¹³C. Respiration of roots and shoots of individual plants was measured with an open CO₂ exchange system interfaced with a mass spectrometer. Respiratory CO₂ from shoots was always ¹³C‐enriched relative to that of roots. Conversely, shoot biomass was always ¹³C‐depleted relative to root biomass. The δ‐difference between shoot and root respiratory CO₂ was variable, and negatively correlated with the δ‐difference between shoot and root biomass (r² = 0.52, P = 0.023), suggesting isotope effects during biosynthesis. ¹³C discrimination in respiration (R) of shoots, roots and whole plants (e_Shoot, e_Root, e_Plant) was assessed as e = (δ_Substrate − δ_R)/(1 + δ_R/1000), where root and shoot substrate is defined as imported C, and plant substrate is total photosynthate. Estimates were obtained from C isotope balances of shoots, roots and whole plants of sunflower and alfalfa using growth and respiration data collected at intervals of 1 to 2 weeks. e_plant and e_Shoot differed significantly from zero. e_plant ranged between −0.4 and −0.9‰, whereas e_Shoot was much greater (−0.6 to −1.9‰). e_Root was not significantly different from zero. The present results help to resolve the apparent conflict between leaf‐ and ecosystem‐level ¹³C discrimination in respiration.

Leaf growth in monocotyledons results from the flux of newly born cells out of the division zone and into the adjacent elongation-only zone, where cells reach their final length. We used a kinematic method to analyze the effect of phosphorus nutrition status on cell division and elongation parameters in the epidermis of Lolium perenne. Phosphorus deficiency reduced the leaf elongation rate by 39% due to decreases in the cell production rate (−19%) and final cell length (−20%). The former was solely due to a lower average cell division rate (0.028 versus 0.046 cell cell−1 h−1) and, thus, a lengthened average cell cycle duration (25 versus 15 h). The number of division cycles of the initial cell progeny (five to six) and, as a result, the number of meristematic cells (32–64) and division zone length were independent of phosphorus status. Accordingly, low-phosphorus cells maintained meristematic activity longer. Lack of effect of phosphorus deficiency on meristematic cell length implies that a lower division rate was matched to a lower elongation rate. Phosphorus deficiency did not affect the elongation-only zone length, thus leading to longer cell elongation duration (99 versus 75 h). However, the substantially reduced postmitotic average relative elongation rate (0.045 versus 0.064 mm mm−1 h−1) resulted in shorter mature cells. In summary, phosphorus deficiency did not affect the general controls of cell morphogenesis, but, by slowing down the rates of cell division and expansion, it slowed down its pace.

Summary: Carbon (C) allocation strongly influences plant and soil processes. Short‐term C allocation dynamics in ecosystems and their responses to environmental changes are still poorly understood. Using in situ ¹³CO₂ pulse labeling, we studied the effects of 1 wk of shading on the transfer of recent photoassimilates between sugars and starch of above‐ and belowground plant organs and to soil microbial communities of a mountain meadow. C allocation to roots and microbial communities was rapid. Shading strongly reduced sucrose and starch concentrations in shoots, but not roots, and affected tracer dynamics in sucrose and starch of shoots, but not roots: recent C was slowly incorporated into root starch irrespective of the shading treatment. Shading reduced leaf respiration more strongly than root respiration. It caused no reduction in the amount of ¹³C incorporated into fungi and Gram‐negative bacteria, but increased its residence time. These findings suggest that, under interrupted C supply, belowground C allocation (as reflected by the amount of tracer allocated to root starch, soil microbial communities and belowground respiration) was maintained at the expense of aboveground C status, and that C source strength may affect the turnover of recent plant‐derived C in soil microbial communities.

Patterns of synthesis and breakdown of carbon (C) and nitrogen (N) stores are relatively well known. But the role of mobilized stores as substrates for growth remains less clear. In this article, a novel approach to estimate C and N import into leaf growth zones was coupled with steady-state labeling of photosynthesis (¹³CO₂/¹²CO₂) and N uptake (¹⁵NO₃⁻/¹⁴NO₃⁻) and compartmental modeling of tracer fluxes. The contributions of current C assimilation/N uptake and mobilization from stores to the substrate pool supplying leaf growth were then quantified in plants of a C₃ (Lolium perenne) and C₄ grass (Paspalum dilatatum Poir.) manipulated thus to have contrasting C assimilation and N uptake rates. In all cases, leaf growth relied largely on photoassimilates delivered either directly after fixation or short-term storage (turnover rate = 1.6–3.3 d⁻¹). Long-term C stores (turnover rate < 0.09 d⁻¹) were generally of limited relevance. Hence, no link was found between the role of stores and C acquisition rate. Short-term (turnover rate = 0.29–0.90 d⁻¹) and long-term (turnover rate < 0.04 d⁻¹) stores supplied most N used in leaf growth. Compared to dominant (well-lit) plants, subordinate (shaded) plants relied more on mobilization from long-term N stores to support leaf growth. These differences correlated well with the C-to-N ratio of growth substrates and were associated with responses in N uptake. Based on this, we argue that internal regulation of N uptake acts as a main determinant of the importance of mobilized long-term stores as a source of N for leaf growth.

The practice of inoculating forage legumes with rhizobia strains is widespread. It is assumed that the inoculated strain determines the performance of the symbiosis and nitrogen fixation rates. However, native-naturalized strains can be competitive, and actual nodule occupancy is often scarcely investigated. In consequence, failures in establishment, and low productivity attributed to poor performance of the inoculant may merely reflect the absence of the inoculated strain in the nodules. This study lays out a strategy followed for selecting a Rhizobium leguminosarum sv. trifolii strain for white clover (Trifolium repens) with competitive nodule occupancy. First, the competitiveness of native-naturalized rhizobia strains selected for their efficiency to fix N₂ in clover and tagged with gusA was evaluated in controlled conditions with different soils. Second, three of these experimental strains with superior nodule occupancy plus the currently recommended commercial inoculant, an introduced strain, were tested in the field in 2 years and at two sites. Plant establishment, herbage productivity, fixation of atmospheric N₂ (¹⁵N natural abundance), and nodule occupancy (ERIC-PCR genomic fingerprinting) were measured. In both years and sites, nodule occupancy of the native-naturalized experimental strains was either higher or similar to that of the commercial inoculant in both primary and secondary roots. The difference was even greater in stolon roots nodules, where nodule occupancy of the native-naturalized experimental strains was at least five times greater. The amount of N fixed per unit plant mass was consistently higher with native-naturalized experimental strains, although the proportion of N derived from atmospheric fixation was similar for all strains. Plant establishment and herbage production, as well as clover contribution in oversown native grasslands, were either similar or higher in white clover inoculated with the native-naturalized experimental strains. These results support the use of our implemented strategy for developing a competitive inoculant from native-naturalized strains.