Nitrogen acquisition, fixation and transfer in maize/alfalfa intercrops are increased through root contact and morphological responses to interspecies competition

Nitrogen (N) fixation by legumes and nitrogen transfer to cereals have been considered as important pathways for overyielding and higher N use efficiency in cereal/legume intercropping systems. However, the extent to which root morphology contributes to N fixation and transfer is unclear. A two-factorial greenhouse experiment was conducted to quantify the N fixation, transfer and root morphology characteristics of the maize/alfalfa intercropping system in two consecutive years using the 15N-urea leaf labeling method, and combining two N levels with three root separation techniques. N application could inhibit N fixation and transfer in a maize/alfalfa intercropping system. Irrespective of the N application level, compared with plastic sheet separation (PSS), no separation (NS) and nylon mesh separation (NNS) significantly increased the total biomass (36%) and total N content (28%), while the N fixation rate also sharply increased by 75 to 134%, and the amount of N transferred with no root barrier was 1.24–1.42 times greater than that with a mesh barrier. Redundancy analysis (RDA) showed that the crown root dry weight (CRDW) of maize and lateral root number (LRN) of alfalfa showed the strongest associations with N fixation and transfer. Our results highlight the importance of root contact for the enhancement of N fixation and transfer via changes in root morphology in maize/alfalfa intercropping systems, and the overyielding system was achieved via increases in maize growth, at the cost of smaller decreases in alfalfa biomass production.