Fig. 3: The spectrum of phenotypes seen in SUP hypermethylated diploid natural accessions.

a A representative Geg-14 accession inflorescence depicting a spectrum of sup phenotypes: superwoman (blue arrowhead), superman (red arrow), and curly siliques(red asterisk). b A flower from Geg-14 inflorescence with supersex phenotype. c Dissected third and fourth whorls of the flower b respectively showing nine anthers and three carpels. d A flower from Geg-14 inflorescence with superman phenotype e dissected third and fourth whorls of the flower shown in d, respectively, showing seven anthers and a pistillode (sterile pistil) as indicated by arrowheads. f Replum–septum skeletons of dehisced silique from Geg-14 showing multiple replums (red arrows) and abnormal fusion of septum (blue arrowheads). g A partial tricarpellary silique from IP-Trs-1 plant. h The silique shown in g is split open to reveal its anatomy. The silique is bicarpellary and normal (red arrow) at the start, however in the midway, one of the repla bifurcates (blue arrowhead) to produce a partial tricarpellary silique (three carpel valves are shown). i–k The spectrum of replum–septum skeleton phenotypes from various accessions (as indicated) that show weak superwoman phenotypes. l Replum-septum skeleton from the SUP silent Basta-2 accession showing WT phenotypes. m Chop PCR assay in the natural accessions with hypermethylated SUP locus using the McrBC enzyme. The strong lol epialleles from Wa-1, Geg-14, and the moderate lol epialleles from IP-Trs-0 and the weak lol epiallele from Got-22 are fully methylated at cytosines at McrBC recognition sites, as they are PCR negative for the primers flanking patch-2. The remaining accessions show a faint PCR signal compared to their respective undigested controls suggestive of heterogeneous methylation of cytosines at this site. n Chop PCR assay in the natural accessions using the Dde1 enzyme. The strong lol epialleles from Wa-1, Geg-14, and the weak lol epialleles from Got-22 and Sorbo contain either fully or partially methylated cytosines at Dde1 recognition sites, as they are PCR positive for the primers flanking patch-2. The remaining accessions are PCR negative suggestive of the absence of cytosine methylation at this site. DNA markers used in the gels is 50 bp NEB ladder. Scale bars: a = 1 cm, b–l = 1 mm.