Effect of deleption of BnaA06FAH and BnaC05FAH on gene expression during programmed cell death in brassica napus.L
Programmed cell death (PCD) is a genetically controlled pathway that plants can use to selectively eliminate redundant or damaged cells.The coordinated and sequential actions of lineage-specific transcription factors and epigenetic regulators are essential for the initiation of PCD. We used CRISPR/Cas9-mediated targeted mutagenesis to generate a series of transgene-free mutant lines either with single or double-null bnafah alleles. Among these mutant lines, the bnafah (deleption of BnaA06FAH and BnaC05FAH) double-null mutant line, rather than the bnaa06fah (deleption of BnaA06FAH) mutant line, exhibited PCD under short day (SD) condition. To gain deeper mechanistic understanding of PCD process, transcriptomic analyses of bnaa06fah and bnafah under LD(SD0), SD1 and SD2 were performed. Heatmap cluster of DEGs speculated that PCD may related to various redox regulation genes involved in antioxidant, ROS-responsive regulator and calcium signaling. Combined with the results of previous studies, our work showed that the expression levels of BnaC04CAT2, BnaA09/C09SAL1, BnaA08/C08ACO2, BnaA07/C06ERO1, BnaA08ACA1, BnaC04BIK1, BnaA09CRK36 and BnaA03CPK4 were significantly different and might be candidate hub genes of PCD. Overall design: To investigate deleption of both BnaA06FAH and BnaC05FAH or single BnaA06FAH in regulating programmed cell death, We used CRISPR/Cas9-mediated targeted mutagenesis to generate a series of transgene-free mutant lines either with single or double-null bnafah alleles. To gain deeper mechanistic understanding of PCD process, transcriptomic analyses of bnaa06fah (deleption of BnaA06FAH) and bnafah (deleption of BnaA06FAH and BnaC05FAH) under different SD stages were performed.