We have described a new unstable mutant of the vestigial locus isolated from a natural population. From this mutant, vestigial(almost) (vg(al)), wild-type (vg(al+)), and extreme (vg(ext)), alleles arose spontaneously. The molecular analysis of vg(al) shows that the mutation is due to a 1874 bp hobo element inserted in a vestigial intron. Two distinct kinds of events lead a wild-type phenotype. Three independent vg(al+) alleles result from an excision of the hobo element and two other vg(al+) alleles have further deletions of hobo sequence. The sequence of one of them shows a 1516 bp hobo insertion at the same place and in the same orientation as the 1874 bp insertion. In the vg(ext) alleles, we found a 5' or 3' variably sized deletion of vg sequences. One of them, which has been cloned and sequenced, has a deletion finishing exactly at the left terminal repeat' hobo element. The genetic implications of these different genetic structures are discussed.