The development of varieties is a continuous process and the success of the plant breeding depends upon the selection of suitable plants to be utilized. The effectiveness of selection depends basically upon the magnitude of genetic variability in the breeding material. This study was carried out at Africa Rice Center, International Institute of Tropical Agriculture (IITA) Ibadan, Nigeria. Two hundred and thirty-nine (239) lines of anther-culture derived from South Korea with an improved variety from Nigeria as check were established for their genetic variability and diversity analysis. The experiment was conducted using Alpha lattice design with four blocks each planted in 60 entries replicated tow times. The estimation of genotypic coefficient variance and phenotypic coefficient variance was found to be high (>20%) for grain yield, grain yield per plant, biomass, number of tillers, panicle weight, effective tillers, leaf area, leaf area index and number of grains per panicle. The broad sense heritability was highest for days to 50% flowering followed by plant height, 1000 grain weight, panicle length and number of tillers. The estimation of genetic advance was found to be highest for grain yield. Cluster analysis grouped the 240 accessions into four clusters (A, B, C and D), indicate wide genetic diversity among these groups. Principal component analysis showed that the first three components accounted for 64.78% of the total variation. Therefore, indicate the presence of large genetic variability, which is important as it gives wide range of selection. Among all genotypes UPN 632 and UPN 540 showed the best performance.