The comparison for the transcript levels of chloroplast-encoded genes between wild type (Col-0) and pbf8 mutants
We found that thylakoid-anchored protein PBF8 is a key regulator for Photosystem I (PSI) biogenesis. To explore the role of PBF8 in regulating chloroplast gene expression, we performed the RNA-seq to compare the the transcript levels of chloroplast-encoded genes between wild type (Col-0) and pbf8 mutants. To this end, we isolated the total RNA form 12-day-old wild type and pbf8 seedlings grown on the MS medium under long-day conditions (14 h light, 10 h dark) at 22 ºC and with a light intensity of 80 µmol m-2 s-1. The rRNAs were deleted using the Ribo-Zero Kit (Epicentre). The resulting rRNA-depleted RNA was used for preparing the sequencing library with NEBNext Single Cell/Low input library Prep Kit. The libraries were pooled and sequenced on an Illumina Nova 6000 system with 150-bp pair-end reads. Finally, our results show that the transcript accumulation for chloroplast-encoded PSI subunit and assembly factor genes between the wild type (Col-0) and pbf8 samples, suggesting PBF8 may not affect the transcript levels of chloroplast-encoded PSI subunits and assembly factors in chloroplasts. Overall design: To investigate the role of PBF8 in regulating chloroplast gene expression, we obtained the pbf8 T-DNA insertion mutant in which PBF8 gene has been knocked out. We then performed gene expression profiling analysis using the data form RNA-seq of wild type and pbf8 seedlings. Comparative profiling analysis of chloroplast-encoded genes of RNA-seq data for wild type and pbf8 seedlings.