Table 1

Primers used in this study"

Fig. 1

Schematic diagram of OsCDF1 gRNA target sites and screening for positive clones containing the gRNAs of OsCDF1 A: OsCDF1 target site in schematic diagram. The grey, blue rectangles, and black lines represent 5' or 3' UTRs, exons and introns, respectively. Bar: 200 bp. PAM: protospacer adjacent motif. The sequences underlined indicate PAM sequences. B: the amplification of the fragments containing gRNAs in pRGEB32 vector by PCR. M: DNA marker III; 1-2: PCR products. C: the sequencing results of the positive clones containing gRNAs of OsCDF1."

Fig. 2

Screening and analysis of oscdf1 homozygous mutants without T-DNA A: PCR amplification of Hyg fragments. M: DNA marker III; 1-48: PCR products; 1: template is ddH2O. B: mutation type analysis in T1 generation. C: the mutation type analysis in T2 generation. Red arrows show the mutation sites. D: the change of the target sites. The grey, blue rectangles, and black lines represent 5' or 3' UTRs, exons, and introns, respectively. PAM: protospacer adjacent motif. The sequences underlined represent PAM. Bar: 200 bp."

Fig. 3

Schematic representations of amino acid sequence change and tertiary structure analysis of OsCDF1 mutant proteins A: the amino acid change of the target sites. Bar: 50 aa. B: the blast analysis of the mutant proteins. C: the tertiary structure."

Fig. 4

Flowering time phenotype of oscdf1 mutants A: phenotype of oscdf1 mutants. Bar: 10 cm in white line. B: flowering time of oscdf1 mutants (n ≥ 15). *: P < 0.05."

Fig. 5

Comparison of agronomic traits between oscdf1 mutants and wild-type grown in paddy field A: the morphology of panicle between WT and oscdf1 mutant; B: the morphology of branch between WT and oscdf1 mutant; C: panicle number; D: panicle length; E: primary branch number; F: secondary branch number; G: grain length. H: grain width; I: grain length; J: grain width; K: grain thickness; L: the filled grain number per panicle; M: seed-setting rate; N: 1000-grain weight; O: yield weight per plant. Data in C-F and I-J are means ± SDs (n ≥ 15). *: P < 0.05; **: P < 0.01; n.s.: no significant difference."

Fig. 6

Expression pattern of the OsCDF1 gene A: tissue-specific analysis of OsCDF1. B: the relative expression level of OsCDF1 different developmental stages. Data in A and B are means ± SDs."

Fig. 7

Genetic variation and haplotype network analysis of OsCDF1 A: OsCDF1 gene structure and location of the six genomic variations. Black rectangles represent the two exons; Grey rectangles represent the five' untranslated region (5' UTR) and 3' UTR; Black lines represent the intron. B: RFT1 protein structure and location of the six genomic variations. Triangles in different colors indicate the locations of the genetic variations. C: the haplotype network of the six genomic variations. The red lines represent the number of mutations between two haplotypes."