Table 1.
Regeneration pathway | Target material (explants) | Pre-culture | Agrobacterium tumefaciens strain | Transferred genes | Selection | Transformation efficiency | Reference |
---|---|---|---|---|---|---|---|
Adventitious shoot formation | Etiolated petioles | 6 days | AGL-0, LBA4404 | gusAint, hpt & nptII | 5 mgL−1 hyg or 100 mgL−1 kan | 9–19% (AGL-0); 0–2.5% (LBA4404) | Aida et al., 1999 |
Adventitious shoot formation | Etiolated hypocotyl segments | 1–8 days | LBA4404, EHA105 | nptII, gusAint | 50 mgL−1 kan | 15.3% (LBA4404); 2.3% (EHA105) | Boase et al., 2002 |
Secondary somatic embryogenesis | Somatic embryos | 3 months | LBA4404 | gusAint, hpt | 5 mgL−1 hyg | 0.7% hygR calluses | Terakawa et al., 2008 |
Adventitious shoot formation | Etiolated hypocotyl segments | 1–8 days | EHA105 | CpF3′5′H hpt | 5 mgL−1 hyg (12 days); 20 mgL−1 hyg (65 days); 15 mgL−1 hyg (thereafter) | Not indicated | Boase et al., 2010 |
Adventitious shoot formation | Etiolated petioles regenerated from leaf explants | – | LBA4404 | CpTCP-SRDX, hpt | 5 mgL−1 hyg | 53% | Tanaka et al., 2011 |
Adventitious shoot formation | Etiolated petioles of seedlings | 6 days | EHA105 | CpFAD7, hpt | 10 mgL−1 hyg | Not indicated | Kai et al., 2012 |
Adventitious shoot formation | Etiolated petioles regenerated from leaf explants | – | GV3101, LBA4404 | CpAG1-SRDX, CpAG2-SRDX, hpt | 5 mgL−1 hyg | Not indicated | Tanaka et al., 2013 |
Somatic embryogenesis | Embryogenic callus | 10 days | LBA4404, EHA105, GV2260, GV3101, AGL-1 | hpt, gusAint, DR5::gus, roGFP2_Orp1, | 5–10 mgL−1 hyg (14 days), 10–20 mgL−1 (thereafter) sensitivity depends on genotype | 43% (EHA105), 20% (LBA4404) | Ratjens et al., this study |
gusAint, reporter gene gusA containing an intron; hpt, hygromycin (hyg) phosphotransferase gene; nptII, neomycin phosphotransferase gene conferring kanamycin (kan) resistance; CpF3′5′H, flavonoid 3′5′-hydroxylase of Cyclamen persicum; CpFAD7, omega-3 fatty acid desaturase of Cyclamen persicum; CpTCP-SRDX, Cyclamen persicum TCP transcription factor – repressor; CpAG-SRDX, Cyclamen persicum AGAMOUS – repressor; hygR, hygromycin-resistant cells growing on hygromycin-containing medium; DR5, synthetic auxin response element; roGFP2_Orp1, redox-sensitive green fluorescent protein biosensor version 2 integrated with a yeast peroxidase gene.