Fig. 5.

Effect of BmDnmt2 on RPC and silk yield traits. a Detection of mutant alleles in the BmDnmt2-KO genomic sequence. The gene structure of BmDnmt2 is shown at the top. The two sgRNA sites (sgR1 and sgR2) of BmDnmt2 are marked in red, and the 178 bp deletion of BmDnmt2-KO between the two sites is shown in the middle. The sequencing results are shown below. At the bottom is the result of the PCR amplification of the deletion sequence in the wild-type and BmDnmt2-KO line. b Detection of mutant alleles in the BmDnmt2-KO coding sequence. There were 74 bp between the two sites in the wild-type, an 86 bp deletion in BmDnmt2-KO between the two sites in the coding sequence and a stop codon (TGA) was introduced. The results of base sequencing and gel electrophoresis are shown below. c Schematic diagram of structural changes in the BmDnmt2-KO protein. The brown box is the DNA methylase domain (PF00145.19). d Phenotypic investigation of RPC, CSW, CW, and PW in females (left) and males (right) of wild-type (WT) and BmDnmt2-KO (KO), respectively. Sample size is shown in parentheses following WT and KO, respectively. Center line, median; white dots, mean; box limits, upper and lower quartiles; whiskers, 1.5 × the interquartile range; black dots, outliers (**P < 0.01, ***P < 0.005, two-sided Wilcoxon test)