The maize Ac/Ds transposon family was the first transposable element system identified and characterized by Barbara McClintock. Ac/Ds transposons belong to the hAT family of class II DNA transposons. We and others have shown that Ac/Ds elements can undergo a process of alternative transposition in which the Ac/Ds transposase acts on the termini of two separate, nearby transposons. Because these termini are present in different elements, alternative transposition can generate a variety of genome alterations such as inversions, duplications, deletions, and translocations. Moreover, Ac/Ds elements transpose preferentially into genic regions, suggesting that structural changes arising from alternative transposition may potentially generate chimeric genes at the rearrangement breakpoints. Here we identified and characterized 11 independent cases of gene fusion induced by Ac alternative transposition. In each case, a functional chimeric gene was created by fusion of two linked, paralogous genes; moreover, each event was associated with duplication of the 70-kb segment located between the two paralogs. An extant gene in the maize B73 genome that contains an internal duplication apparently generated by an alternative transposition event was also identified. Our study demonstrates that alternative transposition-induced duplications may be a source for spontaneous creation of diverse genome structures and novel genes in maize.KEYWORDS Ac/Ds alternative transposition; translocation; segmental duplication; chimeric gene; exon shuffling T HE complex architecture of the modern maize genome was shaped by ancient whole-genome duplication and subsequent diploidization events (Messing et al. 2004;Haberer et al. 2005). Transposable elements proliferated during the diploidization process (Gaut et al. 2000) and today comprise 85% of the maize genome (Schnable et al. 2009). For example, multiple insertions of high-copy-number families of class I retrotransposons greatly expanded the intergenic regions of the maize genome. Comparisons of maize and its close relative sorghum indicate that gene number and colinearity are largely conserved (Hulbert et al. 1990), whereas the intergenic regions differ greatly due to frequent retrotransposon insertions in maize. A good example is the adh1 region of maize inbred B73. Nineteen nested LTR retroelements and two solo LTRs together make up 74% of the intergenic sequences between adh1 and the nearest adjacent gene (Tikhonov et al. 1999). Transposition and proliferation of retroelements thus can largely explain genome size variations, the C-value paradox, and discordant evolutionary history among different grass species (SanMiguel and Vitte 2009). Class II DNA transposons generally do not amplify to such high copy numbers as the class I retroelements, but they can induce structural changes in various ways. For example, the maize Ac/Ds transposon family, class II transposable elements, can undergo a process of alternative transposition that can generate large-scale structural changes such a...