Quantifying below-ground nitrogen of legumes -: 1.: Optimising procedures for 15N shoot-labelling

Quantifying below-ground nitrogen (N) of legumes is fundamental to understanding their effects on soil mineral N fertility and on the N economies of following or companion crops in legume-based rotations. Methodologies based on N-15 shoot-labelling with subsequent measurement of N-15 in recovered plant parts (shoots and roots) and in the root-zone soil have proved promising. We report four glasshouse experiments with objectives to develop appropriate protocols for in situ N-15 labelling of the four legumes, fababean (Vicia faba), chickpea (Cicer arietinum), mungbean (Vigna radiata) and pigeonpea (Cajanus cajan). Treatments included N-15-urea concentration (0.1-2.0% w/w), feeding technique (leaf-flap and petiole), leaflet/petiole position (top and bottom of shoot) and frequency of feeding (one and two occasions). N-15-labelling via the leaf-flap was best for fababean, mungbean and pigeonpea, whilst petiole feeding was best for chickpea, in all cases at the lower-stem nodes 3 or 4 using 0.2 mL volumes of 0.5% urea (98 atom% N-15 excess). Fed leaflets and petioles were removed within 2 weeks of labelling. Uneven N-15 enrichment of the nodulated roots because of effects of the less-enriched nodules meant that root derived N in soil would be overestimated if recovered roots were more heavily nodulated than unrecovered roots. One possible solution would be to assume crown nodulation of the plants. Thus, recovered roots would be nodulated; root-derived N remaining in soil may be without nodules. The ratios of nodulated root to unnodulated root enrichments could then be used as an adjustment in the calculations, i.e. in the case of fababean and chickpea, by dividing calculated root-derived N in soil by 1.12 (fababean) and 1.56 (chickpea).