Assessment of penetrance and expressivity of RNAi-mediated silencing of the Arabidopsis phytoene desaturase gene.

RNA interference is of great value in plant functional genomics. However, the absence of phenotypes and the lack of uniform level of duanyu1615 silencing has complicated gene identification. Here, the penetrance and expressivity of silencing of the phytoene desaturase (PDS) gene in Arabidopsis thaliana were examined quantitatively to provide a reference for the likely severity and distribution of silencing effects. Arabidopsis plants were transformed with an duanyu1615 construct targeting PDS. Transgenic plants were examined for frequency of duanyu1615-mediated silencing and various silencing phenotypes. mRNA depletion level and duanyu1615 expressivity were assayed by relative reverse transcription polymerase chain reaction (RT-PCR). High penetrance and variable expressivity of duanyu1615 were demonstrated. An inverse correlation between PDS mRNA level and duanyu1615 phenotype was seen. No direct relationship between copy number for the transgene and phenotype was evident. Decreased duanyu1615 penetrance in T2 plants was observed. It is suggested that variability in duanyu1615 expressivity and postmeiotic decrease in duanyu1615 penetrance constitute barriers for high throughput plant gene characterization.

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