Rice (Oryza sativa) plants acquired excess photosynthates in the form of nonstructural carbohydrates (NSCs) in their stems and grain. Despite keen interest in rice NSC, the dynamics of NSC accumulation, translocation and re-accumulation have not yet been well investigated. AG1 and AG2 QTLs associated with flooding tolerance through catalyzing starch into soluble sugar in germinating seeds. Here we conducted three experiments, greenhouse and field to lay the groundwork for large-scale diversity studies on grain NSC and some agronomic traits under direct-seeded rice (DSR) system, using elite lines incorporating AG1, AG2 and AG1-AG2 QTLs into the popular varieties PSB Rc82 and Ciherang-Sub1 along with the donors Kho Hlan On (AG1) and Ma-Zhan Red (AG2). In germinating seedlings, soluble sugars increased, while starch concentration decreased gradually especially in the tolerant checks and AG1-AG2 introgression lines under flooded soil. Soluble sugar accumulation in stem started to increase from the vegetative stage and peaked at the panicle initiation stage then gradually decreased towards the maturity stage. But Sub1-AG lines had higher sugar and starch concentrations at different growth stages than other genotypes in wet season 2016 and dry season 2017. Plant survival rate was positively correlated with the stem NSC at the early vegetative stage (21 days after sowing), and stem NSC was positively associated with plant height at different growth stages. Among the tested seeding rate, the most suitable seeding rate, 4 g/m2 with shallow burial depth (0.5 cm), resulted in better seedling establishment, relatively higher seedling vigor index and higher leaf area index under flooding in DSR system. Introgression of AG1-AG2 QTLs had no any negative impact on nonstructural carbohydrate, germination rate, and growth and biomass production.