Table 1. Segregation and back-cross segregation analysis of TPXL3 insertion lines.
Genotyping analysis for both TPXL3 insertion lines failed to recover homozygous mutants and segregation analysis showed that both tpxl3-1 and tpxl3-2 lines segregate in a 2:1 (Resistant/Sensitive [R/S]) ratio on antibiotic selection, hinting toward embryo lethality. Back-cross experiments to wild type revealed transmission of the mutant allele via both the male and female gametes, and a reduction in transmission of the tpxl3-1 mutant allele via the female gametes was observed.
| Segregation Analysis | ||||||
|---|---|---|---|---|---|---|
| Line | Insertion | Sensitive | Resistant | Total | Ratio R/S | χ2 |
| tpxl3-1 | SAIL_356B08 | 323 | 680 | 1,003 | 2.1 | |
| Expected (3:1) | 250.8 | 752.3 | 1,003 | 3 | 27.76 | |
| Expected (2:1) | 334.3 | 668.7 | 1,003 | 2 | 0.58 | |
| tpxl3-2 | GK_480B12 | 272 | 546 | 818 | 2.0 | |
| Expected (3:1) | 204.5 | 613.5 | 818 | 3 | 29.71 | |
| Expected (2:1) | 272.7 | 545.3 | 818 | 2 | 0.002 | |
| Back-Cross Segregation Analysis Wild-Type♀ × Mutant♂ | ||||||
| Line | Insertion | Mutant | Wild-Type | Total | Ratio Wild-Type/Mutant | χ2 |
| tpxl3-1 (+/−) | SAIL_356B08 | 73 | 85 | 158 | 1.16 | |
| Expected (1:1) | 79 | 79 | 158 | 1 | 0.91 | |
| tpxl3-2 (+/−) | GK_480B12 | 24 | 18 | 42 | 0.75 | |
| Expected (1:1) | 21 | 21 | 42 | 1 | 0.86 | |
| Back-Cross Segregation Analysis Wild-Type♂ × Mutant♀ | ||||||
| Line | Insertion | Mutant | Wild-Type | Total | Ratio Wild-Type/Mutant | χ2 |
| tpxl3-1 (+/−) | SAIL_356B08 | 44 | 101 | 145 | 2.30 | |
| Expected (1:1) | 72.5 | 72.5 | 145 | 1 | 22.41 | |
| tpxl3-2 (+/−) | GK_480B12 | 46 | 56 | 102 | 1.22 | |
| Expected (1:1) | 51 | 51 | 102 | 1 | 0.98 | |