Aims: The study aimed to evaluate the different growing media & polybag size for seedling growth of rough lemon rootstock. Study Design: The experiment used a Complete Randomized Design (CRD) with three replications. Place and Duration of Study: The study "Effect of Growing Media and Polybag Size on Rough Lemon Seedling Growth" took place in an insect-proof net house at CCS Haryana Agricultural University during the 2021-2022 academic year. Methodology: It comprising of 30 treatment combination i.e., ten different growing media: T1: garden soil; T2: soil: sand (1:1); T3: soil: sand: leafmold (1:1:1); T4: soil: cocopeat (1:1); T5: sand: cocopeat (1:1); T6: soil: sand: cocopeat (1:1:1); T7: soil: FYM (1:1); T8: sand: FYM (1:1); T9: soil: sand: FYM (1:1:1); T10: soil: sand: FYM: cocopeat (1:1:1:1), and the polybags S1: large (30 x 18 cm), S2: medium (21 x 18 cm) & S3: small size (17 x 16 cm) bags. Results: The study found that adding cocopeat to growing media (T4, T5, T6) resulted in earlier germination and higher rates (87-89%) Whereas, T4 to T10 media consists of either FYM, cocopeat for both resulted into more germination 87-89% as compared to controls (T1, T2, T3). The most of the root growth parameters i.e., diameter of tap root(2.45mm), fresh root weight (4.63g) & dry root weight (1.21g) & root: shoot ratio (0.61) improved with the media sand: FYM, soil: FYM, sand: soil: FYM, sand: soil: FYM: cocopeat in equal proportion. However, when media supplemented with cocopeat (T4, T5 & T6 ) also showed appreciable results as compared to soil, sand: soil & soil: sand: leafmold. However, sand: FYM, sand: cocopeat treatments were best in respect of number of primary roots (61.22), root volume (6.44ml). Whereas, sand: soil showed very poor results in all growth aspects. Among polybags sizes, large size (30 x 18cm) bags performed best in most of the germination & root growth parameters. Conclusion: Media enriched with organic matter outperformed sole media across all parameters, while larger polybags produced significantly better results than medium and small sizes.