Journal: PLoS ONE
Article Title: Control of organelle gene expression by the mitochondrial transcription termination factor mTERF22 in Arabidopsis thaliana plants
Figure Lengend Snippet: Transcript abundance of mitochondrial mRNAs in Arabidopsis wild-type and mterf22 plants grown at 28°C. The accumulation of various mtRNAs in wild-type and mterf22 plants grown at 28°C was analyzed by quantitative reverse transcription PCR (RT-qPCR). Total RNA (depleted from DNA) was extracted from 3-week-old wild-type (Col-0) and mutant plants, reverse-transcribed, and the relative steady-state levels of different organellar transcripts were evaluated by qPCR with specific oligonucleotides, after normalization to the actin2 ). The histogram shows the relative mRNAs levels in mutant lines versus those of wild-type plants (i.e., log2 ratios of mutant to wild-type mtRNAs). Transcripts analyzed in these assays include the NADH dehydrogenase (i.e., complex I) subunits nad1 exons a-b, b-c, c-d, d-e, nad2 exons a-b, b-c, c-d, d-e, nad3 , nad4 exons a-b, b-c, c-d, nad4L , nad5 exons a-b, b-c, c-d, d-e, the intronless nad6 subunit, nad7 exons a-b, b-c, c-d, d-e, and nad9 , the complex III cob subunit, cytochrome oxidase (complex IV) cox1 , cox2 exons a-b and cox3 subunits, the ATP synthase (i.e., complex V) subunits atp1 , atp4 , atp6 , atp8 and atp9 , genes encoding the cytochrome c maturation ( ccm ) factors, ccmB , ccmC , ccmFn1 , ccmFn2 and ccmFc exons a-b, the ribosomal subunits rpl2 exons a-b, rps3 exons a-b, rps4 , rps7 , rps12 , rpl16 , rrn26 , and the mttB gene. The values are means of five biological replicates (error bars indicate one standard deviation).
Article Snippet: Quantitative PCR (qPCR) reactions were run on a 384-well LightCycler 480® Instrument II (Roche, Product no. 05015243001), using 2.5 μl of LightCycler 480 SYBR Green I Master mix and 2.5 μM forward and reverse primers in a final volume of 5 μl.
Techniques: Polymerase Chain Reaction, Quantitative RT-PCR, Mutagenesis, Real-time Polymerase Chain Reaction, Standard Deviation