Biological effects of ion beams with different ranges and linear energy transfers (LETs) were investigated in Nicotiana tabacum L. Mature pollen was exposed to doses of 1400 Gy of 6 MeV4He2+ or 1000 Gy of 50 MeV 4He2+, 15 MeV 12C4+, or 460 MeV 40Ar13+ and used to pollinate non-irradiated flowers (I0 generation), resulting in I, seeds. I1 plants were self-pollinated, and mutations were analyzed in the I2 generation. Seed formation in the I0 generation was greatly reduced in all treatments except where 12C4+ irradiated pollen was used. Germination rate and survival rate in the I1 generation were also reduced in the seeds generated from crosses with pollen irradiated with 6 MeV 4He2+ and 12C4+ beams. Furthermore, chromosome and morphological aberrations in I1 plants were observed. However, in 50 MeV 4He2+ and 40Ar13+ regimes, no seeds germinated. In the I2 generation, chlorophyll mutants were very scarce. Morphological mutants were obtained at a frequency of 5.7 × 10−3 and 7.9 × 10−3 in the progeny of 6 MeV 4He2+ and 12C4+ regimes, respectively. It is concluded that the penetration depth is important for inducing mutation and also that pollen can be used for obtaining mutations induced by ion beams with very short penetration depths. Key words: ion beam, mutation, Nicotiana tabacum, pollen.