Drought and water scarcity are significant factors limiting the development of China's maize industry. Deep sowing, as a method to mitigate drought and preserve soil moisture and seedlings, can effectively mitigate the adverse effects of drought stress on their growth. However, deep sowing can influence the emergence of maize seedlings and has a certain impact on their growth and development. To investigate the physiological and transcriptomic changes in maize inbred lines under deep sowing stress, and the alleviation effects of exogenous hormones, this study investigated the response of mesocotyl in deep-seeding tolerant maize inbred line Qi319 and deep-seeding sensitive maize inbred line Zi330 to deep-seeding stress under exogenous MeJA treatment. The physiological and transcriptomic mechanisms underlying the alleviation of deep-seeding stress by exogenous MeJA were elucidated. Results showed that compared to normal sowing depth, mesocotyl length (MES), mesocotyl and coleoptile length (MESCOL) significantly increased under deep-seeding stress for both inbred lines, while seedling length (SDL) was significantly inhibited. However, after adding 1.5 μmol·L-1 MeJA, MES, MESCOL, and SDL significantly increased compared to deep-seeding stress alone. Exogenous MeJA promoted cell elongation, enhanced mesocotyl length, and improved seedling emergence ability. RNA-Seq sequencing revealed differentially expressed genes (DEGs). Under deep-seeding stress compared to normal sowing depth conditions, 1248 DEGs were identified in Qi319 and 1288 DEGs were identified in Zi330. After adding exogenous MeJA under deep-seeding stress compared to normal sowing depth with exogenous MeJA treatment conditions, 538 DEGs were identified in Qi319 and 2284 DEGs were identified in Zi 330. GO enrichment analysis showed that differentially expressed genes were mainly concentrated in biological processes, cell components and molecular functions. KEGG enrichment analysis showed that differentially expressed genes were mainly concentrated in transport and catabolism, signal transduction, amino acid metabolism and lipid metabolism processes. Compared with the deep-seeded treatment, 12 differentially expressed genes were co-expressed in the two inbred lines under exogenous MeJA treatment, and 10 key genes, including ornithine decarboxylase, terpene synthase 7 and ethylene reaction transcription factor 11, were screened out. Our findings elucidate the molecular mechanism underlying maize's tolerance response to deep seeding and offer valuable insights into candidate gene resources for improving deep seeding tolerance in maize.