Induction and Evaluation of Inbreeding Crosses Using the Ant, <em>Vollenhovia Emeryi</em>

The feedback loop of thetransformergene results in proper feminization under multiple sex determination signals in the ant,Vollenhovia emeryi

“…To evaluate whether the HRM assay is applicable to zygosity testing of the two CSD loci, we confirmed that females produced by inbreeding were heterozygous for least one of the two CSD screening regions, and that diploid males were homozygous at both regions. In this experiment, DNA was extracted from whole adults using 5% Chelex 100, as described previously [35]. After that, DNAs (1 µL of each) were used directly in 11.7 µL reaction volumes containing 1 µL of EvaGreenDye (Cosmo Bio Co., Ltd.), 2.5 µL of 5×PrimeSTAR GXL Buffer, 0.25 µL of PrimeSTAR GXL DNA Polymerase, 1 µL of 2.5 mM dNTP, 0.5 µL of each primer (10 µM forward and reverse primers), and 4.95 µL of RNAse-free water.…”

“…Importantly, sexual intermediates never arise, even when feminization (heterozygous CSD) and masculinization (homozygous CSD) primary signals occur simultaneously [28,30]. Although empirical and theoretical studies have shown that ml-CSD systems suppress the cost of inbreeding by reducing the chance that all CSD loci become homozygous [18,32], molecular sex-determination mechanisms controlled by multiple primary signals derived from CSD loci are still unknown.Vollenhovia emeryi is particularly well suited to investigations of molecular sex determination because of its unusual reproductive system, in addition to relative ease of rearing and inbreeding (Figure 2) [33][34][35]. Queens are produced parthenogenetically and are clones of their mothers (Figure 2a,c), while males are produced androgenetically and are genetically identical to their fathers (Figure 2b,e).…”

“…Vollenhovia emeryi is particularly well suited to investigations of molecular sex determination because of its unusual reproductive system, in addition to relative ease of rearing and inbreeding (Figure 2) [33][34][35]. Queens are produced parthenogenetically and are clones of their mothers (Figure 2a,c), while males are produced androgenetically and are genetically identical to their fathers (Figure 2b,e).…”