Fig. 3

Map-based cloning of snfl1. (a) Positional cloning of snfl1. The snfl1 gene was mapped to a 48-kb genomic region between the markers Ind5–3 and RM19157. The marker names are above the vertical lines; the genetic distance and the number of recombinants are displayed below the vertical lines. (b) Ten candidate genes localized between Ind5–3 and RM19157. (c) Gene structure and mutation site of the candidate gene LOC_Os05g50270 (SNFL1). Black boxes indicate exons and grey boxes represent the intron. snfl1 contains one point mutation (G to A) in the last base of the single intron. (d,e) Verification of the mutation site by enzyme digestion. (d) PCR verification of the mutation site before enzyme digestion. (e) PCR verification of the mutation site after enzyme digestion. (f–i) Complementation test of snfl1. (f) Gross morphology of the WT, snfl1 mutant, and complemented transgenic plant (COM) at the grain filling stage. (Scale bar, 20 cm.) (g) Flag leaf of the WT, snfl1 mutant, and COM at the grain filling stage. (Scale bar, 5 cm.) (h,i) Flag leaf length (h) and flag leaf width (i) in the WT, snfl1 mutant, and COM at the grain filling stage. (j) Alignment of DNA and cDNA between the WT and snfl1. Red boxes indicate exons, black box indicates the intron. Black arrows represent the mutation site. The alignment was generated using Vector NTI 10 software