Fig. 2: Cloning and identification of OsSRF8.
From: OsSRF8 interacts with OsINP1 and OsDAF1 to regulate pollen aperture formation in rice
a Fine-mapping of OsSRF8. The molecular markers and numbers of recombinants are noted. OsSRF8 is located on chromosome 2 with seventeen predicted ORFs in the interval between the markers DY-9 and DY-15. LOC_Os02g10110 is identified as the candidate gene. Mutant Ossrf8 harbors (G-A) single substitution results in premature translation termination. ATG, start codon; TAA, stop codon. Confocal images of tetrads of WT (b), the Ossrf8 mutant rescued by genomic OsSRF8 sequence or OsSRF8 fused with eYFP (c, d), Cr-Ossrf8-1 (e) and Cr-Ossrf8-2 (f). APMP is marked with the dashed square. Experiments in (b–f) were repeated three times with similar results. Scale bars, 10 μm. g–k The images of the enlarged APMP regions corresponding to the dashed areas in a-e, respectively. The red dotted lines indicate the boundary of the plasma membrane and the blue dotted lines indicate the outer boundary of the callose wall of microspore at the tetrad stage. Scale bars, 3 μm. Scanning electron microscopy observation of WT (l), the Ossrf8 mutant rescued by genomic OsSRF8 sequence or OsSRF8 fused with eYFP (m, n), Cr-Ossrf8-1 (o) and Cr-Ossrf8-2 (p) mature pollen grains. ≥30 pollen grains were imaged in (l–p) with similar results. Scale bars, 10 μm. q The display of target sites of CRISPR/Cas9. At the target site 1, Cr-Ossrf8-1 sequences exhibit a T (red) base insertion mutation; at the target site 2, Cr-Ossrf8-2 sequences have a 26-bp (red) deletion mutation. Green indicates the 20-bp CRISPR-Cas9 target sequence.
