Abstract: Callus was obtained on modified PRL-4 medium supplemented with 2, 4-D from the segments of the first leaf of wheat seedling which had been germinated for 4 days. Roots and shoots were initiated and complated plantlets thus regenerated. The frequency of callus formation and its growth rate depended upon the concentration of 2, 4-D and the locations of segments on the leaf. When the concentration was 4 ppm or less, the callus could be produced only within the region from the leaf base to about 1cra apart. Both KT and 6-BA were inhibitory to callus formation. Histological examination showed that callus originated from the cells located at the place of vascular bundle sheath. The roots were frequently induced from leaf callus. During the process of callus induction, after 10–14 days incubation, the roots began to appear in the medium containing 2, 4-D of 2 ppm or less. The lower the concentration, the nearer to the base of the leaf the root-forming place was. In contrast, the higher concentration induced rooting at the higher part of the leaf but inhibited at the lower place. After 3 weeks incubation, the highest frequency of root differentiation was about 60%. The callus failed to differntiate in short-period incubation in higher concentration of 2, 4-D, but when it was transferred to lower level, roots could be initiated. Shoot and plantlets were regenerated from callus cultured in several phytohormone combinations and under different conditions. But the frequency of shooting was very low, and some seedling were morphologically abnormal. The nature of shooting is not yet clear and further studies should be carried on. In this paper, causes of failure in the protoplas culture of mesophyll tissue of wheat leaf were also discussed.