A study of 15N transfer between legumes and grasses

The overuse of classical N fertilisers contributes substantially to environmental degradation by pollution of groundwater by nitrates. This leaching of N in waters is also an economic flaw for farmers because only a part of the fertiliser is used by the plants. Here, systems involving mixtures of legumes and grasses represent a sustainable alternative because legumes can fix atmospheric N-2 using symbiotic microbes. N transfer in those mixtures has been thoroughly investigated but little is known concerning the effect of N fertiliser on N transfer between N-fixing legumes and companion grasses. In white clover (Trifolium repens L.) - perennial ryegrass (Lolium perenne L.) associations, N is transferred mostly through rhizodeposition into the soil by clover followed by re-uptake by ryegrass. Rhizodeposition of N occurs through senescence and decomposition of legume tissue or through exudation of N compounds by living cells. Ammonium and amino acids are the main compounds exuded and their exudation is thought to occur by passive diffusion attributed to a concentration gradient from root to soil. In this study, we test the hypothesis that greater N transfer from clover to grass, as seen in N-rich soils or nutrient solutions, is due to greater N rhizodeposition brought about by higher ammonium and amino acid content of roots. The relations between N input, root N content, N net exudation and N transfer between legumes and grasses were investigated using N-15 by growing white clover and perennial ryegrass with increasing N application in axenic microlysimeters or in pots. Ammonium and amino acid concentrations were measured in root tissues, in root bathing solutions and in soils. We found that mineral N application strongly reduced atmospheric N fixation by clover, from 3.0 to 0.9 mg per plant, and root amino acid content, from 164 to 49 nmoles per g dry weight, but had no effect on ammonium and amino acid concentrations in sterile exudates, showing for the first time that amino acid net exudation is independent of root content. In contrast, ammonium and amino acid concentrations in clover soils increased with N fixation, showing the link between N fixation and N rhizodeposition in soils. Nitrate application increased ryegrass root growth by 7 -8 times, and transfer of N between clover and ryegrass (by 3 times). It is concluded that N fertiliser does not modify N exudation but decreases N fixation and ammonium rhizodeposition in soil by clover. N fertiliser increases N transfer between clover and ryegrass by increasing soil exploration by ryegrass and giving a better access to different available N sources, including the N compounds exuded from clover.