Ion implantation, as a new biophysically mutagenic technique, has shown a great potential for horticultural plant breeding. Up to date, little is known about the mutation mechanism of ion implantation at the DNA level. To reveal the mutation effect of Fe+ ion implantation on Baiyangdian red lotus, the random amplified polymorphic DNA (RAPD) was used, and then the bands of mutants and the control in the radiation-sensitive sites were cloned to be sequenced for comparing their DNA sequences. The results indicated that the total base mutation rate of mutants was 0.87%, and there was different in the six mutants. The types of base changes included base transition, transversion, deletion, and insertion. Among the 159 base changes detected, the frequency of single base substitutions (61.01%) was higher than that of base deletions and insertions (38.99%), and the frequency of base transitions (44.65%) was 2.7 times of that of the base transversions (16.35%). The transitions between C and T accounted for largest proportion, A→G transitions and A→T transversions were also present at high frequency. Adenine, thymine, guanine or cytosine could be replaced by any of other three bases, except that there was no C → G substitution. However, thymine was more sensitive to the irradiation than other bases. In our study, we found many purine bases around the purine mutational sites, and many pyrimidine bases around the pyrimidine mutational sites. These will further help us to understand the mechanism of mutagenesis by ion implantation.