Strains | G0 | G1 | G2 |
|---|
Eggs | Hatched (%)# | Adults (%)& | Reciprocal crosses* | Fertile progeny (%)†| Heterozygous (%)‡ | Pupae | Homozygous (%)¶ |
|---|
C2-3KO | 450 | 152/450 (34) | 68/152 (45) | 25 × 25 | 20/50 (40) | 30/70 (43) | 100 | 18/100 (18) |
N1-3aKO | 468 | 173/468 (37) | 75/173 (43) | 25 × 25 | 22/50 (44) | 35/80 (44) | 96 | 24/96 (25) |
- #Among the injected eggs, approximately 34% (152/450) and 37% (173/468) hatched to larvae in C2-3KO and N1-3aKO groups
- &Among the hatched larval, about 45% (68/152) and 43% (75/173) of the larvae developed into adults in G0
- *25 single-pair reciprocal crosses between the G0 progeny and the susceptible DBM1Ac-S were performed to produce G1 offspring
- †Among the 50 single pairs, 20/22 single pairs generate fertile progeny and one or two of the 20/22 single pairs showed mutagenesis in PxABCC3 and PxABCC2 or PxAPN3a and PxAPN1 locus by the specific PCR amplification
- ‡The other larvae from the single pair group with the positive mutation were reared to pupation, the gDNA samples from 70/80 exuviates of the fourth-instar larvae in C2-3KO/N1-3aKO group were used for nondestructive detection, and 30/35 of them were heterozygotes
- ¶18/24 homozygous individuals were determined by direct sequencing using the gDNA samples from 100/96 exuviates of the final fourth-instar larvae in C2-3KO or N1-3aKO group
