Fig. 1

Phenotypic characterization of dek54 mutant A: F2 segregating population of mature ear; B: phenotypes of kernels; C: cross section of kernels; D: longitudinally section of kernels; E: 100-kernel weight, ten-kernel length, and ten-kernel width of WT and dek54 mutant kernels from F2 segregating population. Bar: 0.5 cm. Em: embryo; En: endosperm; SE: starchy endosperm; VE: vitreous endosperm."

Fig. S1

Germination test of wildtype and dek54 mature kernels (5 days after germination) Bar: 1 cm."

Fig. 2

Histological observatin of dek54 mutant A: longitudinal paraffin sections observation of developing WT and dek54 mutant kernels at 16 DAP. Bar: 100 µm. AL: aleurone layer; SG: starch endosperm. B: scanning electron microscopy observation of the central regions of WT and dek54 mutant mature kernel endosperm. Bar: 50 µm. SG: starch granule; PB: protein body."

Fig. 3

Biochemical component analysis of dek54 mutant Protein (A), starch (B), amino acid components (C), and total nitrogen contents (D) of WT and dek54 mutant kernels."

Table 1

Sequences of SSR markers for dek54 mapping"

Table 2

Gene annotation in the mapping region of dek54 "

Fig. 4

Linkage SSR marker screening and fine mapping of dek54 A: the polymorphic PCR products amplified by umc2160 between WP and MP; B: confirmation of linkage SSR marker umc2160 by genotyping 33 dek54 mutant plants; C: fine mapping of dek54 using F2 population including 1566 individuals. dek54 was mapped to an interval about 290 kb flanked by SSR6 and SSR7. The symbols above and below the black vertical solid lines represent the molecular marker and the number of recombinants, respectively."

Fig. 5

Mutation site and spatio-temporal expression analysis of Zm00001d019294 A: gene structure diagram of Zm00001d019294. The red arrow indicates mutation sites. B: the relative expression level of Zm00001d019294 in different tissues and developing immature kernel."

Fig. 6

Phenotypic characterization of CRISPR/Cas9 targeted mutation of Zm00001d019294 A: the sequence in the Zm00001d019294 locus targeted using CRISPR/Cas9. The sgRNA target sequence is green. Protospacer-adjacent motif (PAM) is blue. Red letters and dashes represent insertions and deletions, respectively. B: mature F2 ear of dek54-cas9-1xMo17. C: mature WT and dek54-cas9-1 kernels. D: longitudinal paraffin sections of mature WT and dek54-cas9-1 kernels. Em: embryo; En: endosperm. Bars: 0.5 cm."

Fig. 7

Conserved domain (A) and phylogenetic (B) analysis of Dek54 protein"

Fig. 8

Observation of subcellular localization of Dek54 in maize protoplasts Dek54-GFP was co-localized with a mCherry-labeled plasma membrane marker (mCherry-PM; CD3-1007). Bar: 20 μm."