Aims: Application of carbon (C) and nitrogen (N) isotopes is an essential tool to study C and N flows in plant-soil-microorganisms systems. When targeting single plants in a community the tracers need to be added via e.g., leaf-labeling or stem-feeding approaches. In this study we: (i) investigated if bicarbonate can be used to introduce 14C (or 13C) into white clover and ryegrass, and (ii) compared the patterns of 14C and 15N allocation in white clover and ryegrass to evaluate the homogeneity of tracer distribution after two alternative labeling approaches. Methods Perennial ryegrass and white clover were pulse labeled with 15N urea via leaf-labeling and 14C either via a 14CO2 atm or with 14C bicarbonate through leaf-labeling. Plants were sampled 4 days after labeling and prepared for bulk isotope analysis and for 14C imaging to identify plant parts with high and low 14C activity. Subsequently, plant parts with high and low 14C activity were separated and analyzed for 15N enrichment. Results Bicarbonate applied by leaf-labeling efficiently introduced 14C into both white clover and ryegrass, although the 14C activity in particular for white clover was found predominantly in the labeled leaf. Using 14C imaging for identification of areas with high (hotspots) and low 14C activity showed that 14C was incorporated very heterogeneously both when using bicarbonate and CO2 as expected when using pulse labeling. Subsequent analysis of 15N enrichment in plant parts with high and low 14C activity showed that 15N also had a heterogeneous distribution (up to two orders of magnitude). Conclusion Bicarbonate can efficiently be used to introduce 14C or 13C into plant via the leaf-labeling method. Both 14C and 15N showed heterogeneous distribution in the plant, although the distribution of 15N was more even than that of 14C.
Plant and Soil, 2013, Vol 371, Issue 1-2, p. 191-198