Fig. 4

Determination of mitochondrial DNA content per nuclear DNA and the expression analysis of RpoT genes in tobacco organs using qRT-PCR analysis. Young tobacco leaves, fully expanded leaves, roots, flowers and floral buds were assayed for mitochondrial DNA content relative to nuclear DNA content (A). Primer combination was chosen from mitochondrial intergenic region orf112b to cob (position 34,443–40,865) and from a nuclear non-coding region from RpoTm. Data were determined by calculating the difference of mitochondrial to nuclear CT values. Bars in the figure represent standard deviations for three biological replicates. B RpoTm, and C RpoTmp messenger RNA abundance was determined by quantitative real-time PCR. Data were normalized to cytoplasmic EF1α and GAPDH levels as an internal control. RpoT gene expression is presented as 40-ΔCT values. Given values were derived from three different biological replicates; standard deviations are indicated. Different letters mark mean values that are significantly different from each other (one way ANOVA; p < 0.05 followed by Tukey test)