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<p><em>Rubus fraxinifolius</em> belonging to the wild raspberry group has not been developed in Indonesia. Mutation breeding using gamma-ray as mutagen which was combined with in vitro culture is one of acceleration effort to obtain superior characteristics of the fruit crops, such as larger fruit size, higher nutrition content, plant with less of spines and fruit storage ability. The <em>R. fraxinifolius </em>seeds were irradiated with different doses of gamma-ray ranging from 0 to 500 Gy using the ⁶⁰Co (Cobalt). This research aimed to determine in vitro growth of <em>R. fraxinifolius</em> mutant (M1) after irradiation using gamma-ray. The results showed that the highest percentage of seed germination was obtained on the doses of 100 and 200 Gy. Furthermore, the subculture of <em>R. fraxinifolius </em>mutant (M1) on MS medium with the addition of BA showed the different growth on number of shoots, number of leaves, and plantlet height. Moreover, <em>R. fraxinifolius</em> control showed higher value of shoots, number of leaves, and plantlet height at 4 and 8 weeks after subculture compared to <em>R. fraxinifolius </em>mutant. MS medium with the addition of IBA showed that the number of roots of <em>R. fraxinifolius</em> control (5.75) was higher than that of <em>R. fraxinifolius </em>mutant (M1) (4.83).</p>