Twenty-six upland lines of New Rice for Africa (NERICA) were tested with four Oryza sativa varieties in relation to Fe toxicity tolerance under hydroponic culture containing 1.44 mM Fe (+Fe) and 0 mM Fe (-Fe as a control). Three NERICAs, WAB450-IBP- 24-HB (P24), WAB450-IBP-82-1-1 (P82), and WAB450-IBP-163-3-1 (P163) among the 30 lines/varieties tested possessed relatively strong tolerance judging from reduction of root length and dry weight and shoot dry weight in +Fe compared to -Fe and from iron toxicity score (ITS) in +Fe. Only P24, P82, and P163 showed emerged lateral roots from pericycle at the root elongation zone, whereas in the other 23 NERICAs and four O. sativa varieties lateral root was not observed in the root elongation or differentiation zones. Less disruption of cortex on root tissues was observed in P24, P82, and P163 than in WAB450-16-2-BL2-DV2 (BL2-DV2), the most susceptible NERICA identified. P24, P82, and P163 showed significantly lower Fe content in the shoots than BL2-DV2, suggesting that the tolerant NERICAs could have some mechanism to inhibit the absorption of Fe. The emergence of lateral roots from the root elongation zone in the three tolerant NERICAs would be closely associated with reducing Fe absorption into the plants.