Figure 4
Presence and expression of the gus gene in transformed Fusarium verticillioides isolated from small interfering ( si)RNA-expressing tobacco plants. (A) Polymerase chain reaction (PCR) analysis for the presence of the gus gene in F. verticillioides isolates that were not inoculated in tobacco plants (N), which were isolated from inoculated GUS-RNA interference plants and exhibited a reduction of approximately 62% (RT1) and 96% (RT2) in the gus gene expression and RT2 isolate after reversion to the normal GUS+ phenotype after the 7th passage (R). (B) Reverse transcriptase (RT)-PCR analysis for the presence of transcripts from the endogenous gus gene in fungal cells. (C) RT-PCR analysis for the presence of transcripts from the fungal 5.8S rRNA housekeeping gene (internal control). (D) Detection of siRNA isolated from fungi isolated from transgenic (RT1, RT2 and R) and non-transgenic plants (N). The position corresponding to 18 and 24 nucleotides is indicated. C+: 100 pg of a gus gene-derived oligomer. Ethidium bromide-stained RNA serves as the loading control.