Regulation of olfactory-based sex behaviors in the silkworm by genes in the sex-determination cascade
Fig 5
Loss of BmOR3 expression extends mating time.
(A) Structure of the BmOR3 gene with nine exons indicated by boxes (black boxes, 5’- and 3’-UTRs; white boxes, coding exons). Target sites 1 and 2 are binding sites for sgRNAs. (B) The mean transcript levels (± SEM) of BmOR3 are down-regulated significantly compared to wild-type levels in the three BmOR3 mutant male (M) and female (F) lines. At least five males with mixed antenna were examined for each line. *** indicates p < 0.001 compared with the relevant control using one-way ANOVA. (C) Somatic mutations were induced in the F1 founder animals following crosses of nos-Cas9 with U6-sgRNA strains. PCR analyses with primers to amplify a region of 600 bp revealed deletion mutations in the G0 mutants. The red arrowhead indicates the deleted region. (D) Deletion mutation in the heterozygous offspring after crossing nos-Cas9 and U6-BmOR3sgRNA transgenic silkworm lines. The targeting sequence is shown in black, and the PAM sequence is in red. The deletion size in nucleotides is indicated above the red arrow at the site of the deletion. (E and F) Courtship and mating behavior indexes of BmOR3 mutant and WT males. Data are shown as percentage from 150 pairs tests with chi-squared test. ***p < 0.001. (G) Percentage of autosegregated WT and mutant males *** indicates significant difference at the 0.001 level with chi-squared test.
