Fig. 6
NMR19-4m controls leaf senescence. a Examination of methylation status of NMR19-4 based on Chop-PCR in F1 hybrids and F2 recombinant lines. The arrows at the right side of gel picture indicate the allele that corresponds to the band in the gel. The numbers of A1 to A12 and B1 to B12 in a–c indicate different individual plants and each of them was used for three different assays including Chop-PCR, allele-specific qPCR and darkness-induced leaf senescence. b Examination of expression levels of PPH in NMR19-4m and NMR19-4u alleles by allele-specific qPCR. Error bars are defined as s.e.m (n = 3). c Quantification of chlorophyll content after 5 days of dark treatment in F1 hybrids and F2 recombinant lines. p-value was calculated by a two-tailed t-test, the same below. Error bars are defined as s.e.m (n = 3). d NMR19-4m represses the expression of PPH compared with NMR19-4u in C24 WT background. The different lines correspond to those in Fig. 4c and Supplementary Fig. 9, the same below. We confirmed (Fig. 4; Supplementary Fig. 9) that lines 2, 6, and 13 had unmethylated NMR19-4, and lines 1, 3, and 4 had methylated NMR19-4 in C24 WT background. Error bars are defined as s.e.m. (n = 3). e NMR19-4m delays leaf senescence under dark treatment. Scale bars, 1 cm. f Quantification of the phenotype from e. Total chlorophyll levels were determined before (upper) and after (bottom) 3 d dark treatment. Error bars are defined as s.e.m. (n = 3)
