Mediaa | SH | CC | Stage |
|---|
I | II | III |
|---|
MSb with vitamins (g/L) | 4.4 | 4.4 | 4.4 | 4.4 | 4.4 |
Sucrose (g/L) | 30 | 20 | 20 | 20 | 30 |
pHc | 5.6 | 5.4 | 5.6 | 5.6 | 5.6 |
GA3 (mg/L) | – | – | – | 10 | – |
ZRd (mg/L) | – | 2 | 4 | 4 | – |
NAAe (mg/L) | – | 0.1 | 0.1 | – | – |
Kanamycinf (mg/L) | – | – | 100 | 100 | 100 |
Sulfadiazineg (mg/L) | – | – | – | – | 25 |
Carbenicillinh (mg/L) | – | – | 500 | 500 | 500 |
- aSH: shoot maintenance; CC: co-cultivation; Stage I: callus induction; II: shoot induction; III: root induction
- bMS: Murashige and Skoog [20] with Vitamins (M404, PhytoTechnology Laboratories). GA3: Gibberellic Acid 10 mg/ml stock in ethanol [0.29 mM]; (G745, Sigma-Aldrich) 0.29 μM final concentration
- cThe pH is adjusted to 5.4 or 5.6 with 0.1 N potassium hydroxide before addition of 0.2% Gelzan CM (G3251, PhytoTechnology Laboratories) for solidification
- dZR: Zeatin Riboside 1 mg/ml stock [4.56 mM] (Z875, PhytoTechnology Laboratories); final concentrations 9.1–18.2 μM
- eNAA: 1-napthalene acetic acid (1 mg/ml stock, [5.4 mM] (N605, PhytoTechnology Laboratories); 0.54 μM final concentration
- fKanamycin 50 mg/ml stock; (K378, PhytoTechnology Laboratories) 100 μg/ml final concentration
- gSulfadiazine 25 mg/ml stock; 25 μg/ml final concentration
- hCarbenicillin (C346, PhytoTechnology Laboratories) 500 μg/ml final concentration. Cefotaxime, timentin or vancomycin can be added to Stage I and II in addition to Carbenicillin as required for aggressive Agrobacterium strain
