Table 3.
The effect of exogenous ABA (10 µM exABA), ABA biosynthesis inhibitor (5 µM NOR), inhibitor of auxin transport (10 µM TIBA) and anti-auxin (5 µM PCIB) on the effectiveness of microspore embryogenesis in anther cultures of two DH lines of triticale (× Triticosecale Wittm.) with significantly different androgenic responsiveness (‘responsive’ DH28 and ‘recalcitrant’ DH19)
| Treatment | Days of treatment | ELS/100A | R/100ELS | GR/100ELS | GR/100A | ||||
|---|---|---|---|---|---|---|---|---|---|
| DH19 | DH28 | DH19 | DH28 | DH19 | DH28 | DH19 | DH28 | ||
| Control* | 3 | 3.1 ± 2.0 | 122.8 ± 19.8 | 8.3 ± 8.3 | 13.6 ± 3.4 | 8.3 ± 8.3 | 12.1 ± 3.0 | 0.6 ± 0.6 | 14.4 ± 3.7 |
| 21 | 1.1 ± 0.7 | 97.6 ± 10.4 | 0 | 16.2 ± 3.1 | 0 | 15.4 ± 3.0 | 0 | 13.9 ± 2.6 | |
| exABA | 3 | 7.7 ± 2.8 | 57.9 ± 20.1 | 6.7 ± 6.7 | 6.0 ± 2.3 | 0 | 6.0 ± 2.3 | 0 | 4.6 ± 2.6 |
| 21 | 5.5 ± 1.8 | 98.5 ± 7.8 | 11.0 ± 6.8 | 8.2 ± 1.2 | 10.4 ± 6.9 | 7.2 ± 1.0 | 0.3 ± 0.1 | 7.5 ± 1.5 | |
| NOR | 3 | 4.7 ± 2.4 | 105.8 ± 16.3 | 2.0 ± 2.0 | 8.4 ± 1.6 | 2.0 ± 2.0 | 7.8 ± 1.2 | 0.2 ± 0.2 | 8.1 ± 1.6 |
| 21 | 5.2 ± 1.2 | 109.4 ± 11.7 | 7.1 ± 3.9 | 5.6 ± 1.1 | 6.6 ± 4.0 | 4.9 ± 1.1 | 0.4 ± 0.2 | 5.3 ± 1.3 | |
| TIBA | 3 | 2.5 ± 1.4 | 127.3 | 8.3 ± 8.3 | 11.3 | 8.3 ± 8.3 | 8.7 | 0.4 | 12.9 |
| 21 | 4.2 ± 0.9 | 117.9 ± 10.2 | 11.7 ± 5.7 | 10.7 ± 1.2 | 6.5 ± 3.7 | 9.7 ± 1.2 | 0.3 ± 0.2 | 10.7 ± 1.4 | |
| PCIB | 3 | 4.0 ± 2.0 | 116.7 ± 17.6 | 15.0 ± 12.4 | 12.6 ± 5.2 | 15.0 ± 12.4 | 12.6 ± 5.2 | 0.6 ± 0.4 | 16.7 ± 10.0 |
| 21 | 2.4 ± 1.1 | 102.9 ± 14.8 | 4.7 ± 3.9 | 10.0 ± 2.4 | 0 | 9.1 ± 2.2 | 0 | 8.6 ± 1.8 | |
| DMSO | 3 | 5.9 ± 2.4 | 112.2 ± 13.3 | 0 | 4.6 ± 1.4 | 0 | 4.6 ± 1.4 | 0 | 5.6 ± 2.0 |
| 21 | 4.8 ± 1.0 | 81.2 ± 7.3 | 12.7 ± 4.6 | 11.4 ± 0.7 | 9.1 ± 4.3 | 10.9 ± 0.9 | 0.4 ± 0.2 | 8.8 ± 0.9 | |
All compounds were supplemented to Hogland’s medium at the start of cold tillers treatment (21 days at 4 °C) or 3 days before anthers isolation. DMSO (PCIB solvent) in 0.1 % concentration was used as additional control
Data are the means of 5–7 biological replications ±SE
Each Petri dish containing 100 anthers from a different spike was assumed to be one biological replication
Statistical analysis separate for each parameter and treatment; data marked in bold differ significantly in comparison with control according to non-parametric Kolmogorov–Smirnov test (K–S), p ≤ 0.05
Final efficiency of androgenesis was estimated by GR/100A—the number of green regenerants per 100 isolated anthers
ELS/100A the number of embryo-like structures produced per 100 anthers of the donor plant, R/100ELS total number of regenerants per 100 embryo-like structures transferred to the regeneration medium, GR/100ELS the number of green regenerants per 100 embryo-like structures transferred to regeneration medium, ABA abscisic acid, NOR norflurazone, TIBA 2,3,5-triiodobenzoic acid, PCIB p-chlorophenoxyisobutyric acid
* Cultures of anthers excised from tillers kept during whole cold treatment in Hoagland’s medium (21 days of treatment) or transferred 3 days before anthers isolation to fresh Hoagland’s medium (3 days of treatment)