To elucidate the physiological and metabolic mechanism of perennial grass responses to alkali stress, we selected Leymus chinensis (L. chinensis), a salt-tolerant perennial rhizomatous species of the family Poaceae as experimental material. We conducted a pot experiment in a greenhouse and measured the biomass, physiological characteristics, metabonomic, and corresponding metabolites. Our results showed that alkali stress significantly inhibited seedling growth and photosynthesis, which caused ion imbalance and carbon deficiency, but the alkali stress significantly increased the nitrogen and ATP contents. The metabolic analysis indicated that alkali stress markedly enhanced the contents of nucleotides, amino acids, and organic acids, but it decreased soluble sugar contents. Pathway enrichment analysis showed that the glutamine synthetase/glutamate synthase (GS/GOGAT) cycle, which was related to nitrogen metabolism, was most significantly affected by alkali stress. The contents of glutamine synthetase (GS) and glutamate synthetase (GOGAT) involved in this pathway were also significantly increased. Our results not only verified the important roles of some amino acids and organic acids in resisting alkali stress, but also further proved that nucleotides and the GS/GOGAT cycle related to nitrogen metabolism played critical roles for seedlings in response to alkali stress.