Figure 6

Validation of excision and reinsertion events of mPing by locus-specific PCR amplification in 32 random S1 individuals derived from multiple etoposide-treated S0 plants of RZ1 (in which no evidence for either excision or reinsertion events of mPing was detected) and confirmation of mPing stability in two generations (S0 and S1) of the wild-type control plants of RZ1. (a) The five mPing-containing loci in untreated control of RZ1 showing excisions in the various S1 individuals (depending on locus). Representatives of both the upper- and lower-bands were sequenced, which verified that length difference between the two bands for a given locus was exactly 430, i.e., the full length of mPing, as indicated on the right side of the figure. (b) Five of the 20 characterized mPing-empty loci in untreated control of RZ1 showing reinsertions in the various S1 individuals (depending on locus). Representatives of both the upper- and lower-bands were sequenced, which verified that the length difference between the two bands for a given locus was exactly 430, i.e., the full length of mPing, as indicated on the right side of the figure. (c) and (d) are amplification results by the same five mPing-containing loci and five mPing-devoiding loci as in (a) and (b), respectively, on 40 individual plants of the wild-type control plants of RZ1 at two generations (S0 and S1). Labeling of the band sizes is the same as in (a) and (b).