Grafting improves nitrogen efficiency and stabilizes yield and quality of cucumber by enhancing the NO3 - uptake.

Grafting can promote the growth and nitrogen use efficiency (NUE) of cucumber seedlings under reduced nitrogen (N) application, however, its underlying mechanisms and effects on mature plants remain unknown. For this purpose, self-grafted and rootstock-grafted cucumber plants were treated with two N levels (7 and 4 mM) throughout the entire growth period. The long-term reduced-N treatment significantly limited the growth, root morphology, nitrate (NO₃ ^-) uptake, NUE traits, photosynthesis, phenylalanine ammonia-lyase (PAL) activity, yield, and fruit quality of self-grafted plants but had no influence on rootstock-grafted plants, it even improved their NUE traits, total phenolic and flavonoid contents, and PAL activity. Furthermore, the expression of the NRT1.2, NRT1.5, NRT2.2, and NRT2.5 genes were significantly down-regulated in self-grafted plant roots, while they and the transcription factors NLP6 and LBD38 were up-regulated in rootstock-grafted plant roots under reduced-N environments. Correlation analysis showed that plant growth, root surface area, N-accumulation, N-uptake efficiency (NUpE), NUE, photosynthesis, PAL activity, yield, and fruit quality were all positively correlated with each other; meanwhile, the root morphology, NRT1.2 and NRT2.1 gene expression were all positively correlated with NUpE and NUE. The results demonstrate that under reduced-N application, rootstock grafting can enhance NO₃ ^- uptake and N accumulation to improve the NUE of cucumber plants and resist reduced-N environment through secondary metabolism, maintaining growth, photosynthesis, yield, and fruit quality without adverse effects. The up-regulation of NRT genes and related transcription factors regulates the NO₃ ^- uptake in rootstock-grafted plants. Rootstock grafting will be beneficial for fertilizer conservation and efficient cucumber production. yield and fruit quality.