Fig. 6: SlSGR1 and SlGgpps2 are directly regulated by NOR-like1, carotenoid accumulation is reduced, and chlorophyll degradation is inhibited in CRISPR/Cas9 nor-like1 mutants.
From: A NAC transcription factor, NOR-like1, is a new positive regulator of tomato fruit ripening
a Carotenoid accumulation was reduced in tomato fruits of nor-like1#1 mutant at B + 3 and B + 10 ripening stages compared with that in WT. b Total chlorophyll content in nor-like1#1 and nor-like1#11 mutants was greater than that of in WT at MG and B + 3 stages. c Expression of SlGgpps2 and SlSGR1 were significantly downregulated both in nor-like1#1 and nor-like1#11 mutants compared with that in WT. Asterisks indicate p < 0.01 (Student’s t-test). d Binding of NOR-like1 to the promoters of downstream target genes SlGgpps2 and SlSGR1. The sequences of the wild-type probes containing the NACRS were biotin-labeled. Competition for NOR-like1 binding was performed with 50× and 500 ×  cold probes containing the wild-type NACRS (indicated with red letters) or mutated NACRS (indicated with green letters). The symbols + or – represent presence or absence respectively. +++ indicates increasing amounts. e ChIP-qPCR assay showing the direct binding of NOR-like1 to the promoter of SlGgpps2 and SlSGR1. Values represent the percentage of DNA fragments in WT fruits that co-immunoprecipitated with anti-NOR-like1 or IgG, or DNA fragments in nor-like1#1 fruit that co-immunoprecipitated with anti-NOR-like1 relative to the input DNA. Bars represent ± SD of three independent replicates. Significant differences (Duncan’s multiple range test, p < 0.05) are indicated in lowercase. f Transient expression assay of NOR-like1 strongly activated the SlGgpps2 and SlSGR1 promoters. Each value is the mean of three biological replicates. Asterisks indicate p < 0.01 (Student’s t-test)
