A field evaluation was conducted to compare ten (10) in vitro regenerated maize genotypes obtained from NACGRAB with conventionally grown maize. The study assessed maize streak severity, root lodging, morpho-agronomic, and yield related traits. The ten in vitro regenerated maize was developed on MS medium supplemented with 0.3 mg/L NAA+3.0 mg/L BAP, while the control maize was planted using conventional methods. The results of the statistical analysis revealed that the maize genotypes aged significantly (p < 0.05) influenced growth and yield traits, except for fresh and dry root weights in the control group. Among the genotypes, POP66 SR/ACR94-YQPM exhibited the highest resistance to root lodging, whereas in vitro regenerated plants demonstrated greater resistance to maize streak severity. TZE COMP3C2 displayed superior early maturity and desirable agronomic traits comparable to TZEE-YPOP STR C4, SAMMAZ 19S-14DT DRD, and POOL ISSR QPMX genotypes. Most growth and yield traits showed strong positive correlations, with the first principal component (Prin 1) explaining 71.09% of the total variation (Eigenvalue: 7.82). In contrast, the eleventh principal component (Prin 11) contributed the least (0.23%, Eigenvalue: 0.03). DTSR-WC, POP66 SR/ACR94-YQPM, TZL COMP4C3, and SAMMAZ 19S-14DT DRD were the most promising genotypes for yield-related traits based on their dry matter content. Remarkably, POP66 SR/ACR94-YQPM produced the highest dry shoot (39.16 g) and root weight (3.81 g) while exhibiting strong resistance to root lodging and maize streak disease. These findings suggest that these genotypes could be valuable for maize breeding programs aimed at improving resistance to maize streak virus and yield.