On the relationship between C and N fixation and amino acid synthesis in nodulated alfalfa (Medicago sativa)

Legumes such as alfalfa (Medicago sativa L.) are vital N2-fixing crops accounting for a global N2 fixation of ~35 Mt N year–1. Although enzymatic and molecular mechanisms of nodule N2 fixation are now well documented, some uncertainty remains as to whether N2 fixation is strictly coupled with photosynthetic carbon fixation. That is, the metabolic origin and redistribution of carbon skeletons used to incorporate nitrogen are still relatively undefined. Here, we conducted isotopic labelling with both 15N2 and 13C-depleted CO2 on alfalfa plants grown under controlled conditions and took advantage of isotope ratio mass spectrometry to investigate the relationship between carbon and nitrogen turn-over in respired CO2, total organic matter and amino acids. Our results indicate that CO2 evolved by respiration had an isotopic composition similar to that in organic matter regardless of the organ considered, suggesting that the turn-over of respiratory pools strictly followed photosynthetic input. However, carbon turn-over was nearly three times greater than N turn-over in total organic matter, suggesting that new organic material synthesised was less N-rich than pre-existing organic material (due to progressive nitrogen elemental dilution) or that N remobilisation occurred to sustain growth. This pattern was not consistent with the total commitment into free amino acids where the input of new C and N appeared to be stoichiometric. The labelling pattern in Asn was complex, with contrasted C and N commitments in different organs, suggesting that neosynthesis and redistribution of new Asn molecules required metabolic remobilisation. We conclude that the production of new organic material during alfalfa growth depends on both C and N remobilisation in different organs. At the plant level, this remobilisation is complicated by allocation and metabolism in the different organs.

Supplying nitrate before bud break induces pronounced changes in nitrogen nutrition and growth of young poplars

Tree nutrient research concentrated on endogenous C and N remobilisation in spring has neglected to acknowledge the possibilities of significant effects of N uptake before bud break, especially on the quality of regrowth and N reserve remobilisation. To investigate this subject, experimental studies were performed on young poplars (Populus tremula × Populus alba, clone INRA 717–1B4) grown with a controlled nutrient supply: (i) without N, ‘control’; (ii) N supplied throughout the course of the experiment, ‘N-supply’; and (iii) N supplied only before bud break, ‘N-pulse’. Results confirm the hypothesis that poplar scions can significantly take up nitrate before bud break, amounting to ~34% of the total N stored the previous year. After bud break, emerging leaves restart the sap flow, which increased nitrate uptake to support the regrowth. N-pulse and N-supply treatments were found to have significant effects shortly after a growth period, i.e. by increasing N content of all tissues (e.g. 37 and 81% in new shoots respectively), leaf area (18 and 29%) and specific leaf area (20 and 35%). Therefore, results confirm the hypothesis that early N supply plays a significant role in the N status and N remobilisation involved in the spring regrowth of young trees.

Effects of a barley (Hordeum vulgare) chromosome 6 grain protein content locus on whole-plant nitrogen reallocation under two different fertilisation regimes

A large fraction of protein N harvested with crop seeds is derived from N remobilisation from senescing vegetative plant parts, while a smaller fraction stems from de novo N assimilation occurring after anthesis. This study contrasts near-isogenic barley (Hordeum vulgare L.) germplasm, varying in the allelic state of a major grain protein content (GPC) locus on chromosome 6. Plant material was grown under both low- and high-N fertilisation levels. The analyses indicated that leaf N remobilisation occurred earlier in high-GPC germplasm under both fertilisation regimes, as indicated by an earlier decrease of total leaf N, chlorophylls, soluble- and membrane-proteins. At the same time, kernel free amino acid levels were enhanced, while leaf free amino acid levels were lower in high-GPC barleys, suggesting enhanced retranslocation of organic N to the developing sinks. Enhanced or longer availability of leaf nitrates was detected in high-GPC varieties and lines, at least under high N fertilisation, indicating that the GPC locus profoundly influences whole-plant N allocation and management. Results presented here, together with data from a recent transcriptomic analysis, make a substantial contribution to our understanding of whole-plant N storage, remobilisation and retranslocation to developing sinks.

The impact of defoliation on nitrogen translocation patterns in the woody invasive plant, Buddleia davidii

The influence of defoliation on nitrogen (N) re-translocation and the source for N remobilisation by the invasive shrub, Buddleia davidii Franch. (buddleia) was determined. Eighty plants were grown over two growing seasons, and half were repeatedly defoliated by removing 66% of their leaf area. During the second season, the N supply was labelled with 15N (10 atom% enrichment), to distinguish the use of stored N (unlabelled) from N taken up by roots (labelled) for growth. Defoliation significantly decreased root (39%) and total biomass (26%). Old leaves were the main source of N for remobilisation which was accelerated and increased (by 50% in the second season) in response to defoliation. In spring, root uptake of N increased by 57% in defoliated plants. Thus, defoliation induced changes in N remobilisation and uptake as compensatory growth increased the demand for N. Continued leaf removal decreased the pool of stored N and caused a significant decline in biomass production, especially in roots (39%) and flowers (31%). This has important implications for the efficacy of defoliation as a control measure, as smaller roots suggest a reduced capacity for uptake of nutrients from the soil and reduced flower production may assist in reducing the invasive spread of the species. These findings clearly show that, although the success of B. davidii is associated, in part, with efficient remobilisation of N from storage, this advantage can be overcome by continued defoliation.

Allocation and Retranslocation of 15N by Maize (Zea mays L.) Hybrids Under Field Conditions of Low and High N Fertility

A field experiment was conducted with high and low N fertility regimes to evaluate root-absorbed 15N, and its subsequent remobilisation in four maize hybrids. In previous experiments, potential yield and patterns of senescence had varied among these hybrids. Individual plants were supplied with 15N at the 12-leaf stage and the uptake of the labelled N and its subsequent partitioning during successive harvests was followed. Grain yield and N accumulation at physiological maturity (PM) were reduced 20 and 40%, respectively, under low N compared with high N fertility. Neither hybrid type nor N fertility level initially affected the amount of lSN absorbed. Maximum 15N content was observed at mid grain-fill and a significant 10% loss of 15N from plants was found at PM. Compared with those under low N, plants cultured under high N maintained higher percentages of 15N in the lower stalk and roots over hybrids and across harvests. Kernels were a sink for remobilised 15N from all vegetative and 'other' reproductive tissues. Patterns of N remobilisation varied among different vegetative tissues. N was remobilised from stalks earlier than from leaves, while N from roots was remobilised throughout the season. Stalk and leaves were the major sources of 15N for kernels (about 45% each) while roots contributed much less. Physiological differences between the hybrid with the highest yield at both fertility levels, and with other hybrids, are discussed.