Table 1 Composition of culture media used in this study for Arabidopsis protoplast regeneration
From: Optimization of protoplast regeneration in the model plant Arabidopsis thaliana
Medium name | Medium composition | Storage | Function | References |
|---|---|---|---|---|
Protoplast Induction Medium (PIM) | Gamborg B5 medium containing vitamins, 20 g/L sucrose, 60 g/L myo-inositol, 2 mg/L 6-BAP, and 0.5 mg/L α-NAA (pH adjusted to 5.8 using 2 M NaOH or 1 M HCl). Medium was sterilized using a 0.2 µm syringe filter | Freshly prepared | Induction of protoplast division | |
Culture Medium A (CMA) | Gamborg B5 medium containing vitamins, 72 g/L d-glucose, 1 mg/L 2,4-D, and 0.15 mg/L 6-BAP (pH adjusted to 5.8 using 2 M NaOH or 1 M HCl). Medium was sterilized using a 0.2 µm syringe filter | Freshly prepared | Induction of protoplast division | [17] |
Protoplast Culture Arabidopsis (PCA) | Gamborg B5 medium containing vitamins, 85 g/L d-glucose, 0.75 g/L MgSO4.7H2O, 0.34 g/L CaCl2, 0.05 g/L l-glutamine, 20 mL/L coconut water, 0.1 mg/L 2-IP, and 0.5 mg/L α-NAA (pH adjusted to 5.8 using 2 M NaOH or 1 M HCl). Medium was sterilized using a 0.2 µm syringe filter | Freshly prepared | Induction of protoplast division; microcallus growth | [21] |
Callus Induction Medium (CIM) | Gamborg B5 medium containing vitamins, 20 g/L sucrose, 2 mg/L 6-BAP, and 0.5 mg/L α-NAA (pH adjusted to 5.8 using 2 M NaOH or 1 M HCl). Medium was sterilized using a 0.2 µm syringe filter | Freshly prepared | Microcallus growth | |
Culture Medium C (CMC) | Murashige and Skoog (MS) medium containing vitamins, 54 g/L d-glucose, 20 g/L sucrose, 0.5 g/L myo-inositol, 0.45 g/L l-glutamine, 0.05 mg/L 2,4-D, and 1 mg/L kinetin (pH adjusted to 5.8 using 2 M NaOH or 1 M HCl). Medium was sterilized using a 0.2 µm syringe filter | Freshly prepared | Microcallus growth | [17] |
Shoot Induction Medium (SIM) | MS medium containing vitamin, 30 g/L sucrose, 0.47 g/L MES.H2O, 0.1576 mg/L IAA, and 0.501 mg/L 2-IP. The pH was adjusted to 5.8 using 2 M NaOH or 1 M HCl before the addition of plant agar (8 g/L), and the medium was sterilized by autoclaving at 121 °C for 10 min | Freshly prepared | Induction of shoot regeneration | [29] |
Shoot Regeneration Medium A (SRMA) | MS medium containing vitamins, 20 g/L sucrose, 0.47 g/L MES.H2O, 7 mg/L 2-IP, and 0.05 mg/L IAA. The pH was adjusted to 5.8 using 2 M NaOH or 1 M HCl before the addition of plant agar (8 g/L), and the medium was sterilized by autoclaving at 121 °C for 10 min | Freshly prepared | Induction of shoot regeneration | [17] |
Shoot Regeneration Arabidopsis (SRA) | Half-strength MS (1/2 MS) medium containing vitamin, 15 g/L sucrose, 0.47 g/L MES.H2O, 2 mg/L kinetin, and 0.05 mg/L IAA. The pH was adjusted to 5.8 using 2 M NaOH or 1 M HCl before the addition of plant agar (8 g/L), and the medium was sterilized by autoclaving at 121 °C for 10 min | Freshly prepared | Induction of shoot regeneration | [23] |
Murashige and Skoog Medium (MS) | 1/2 MS medium containing vitamin, 10 g/L sucrose, and 0.47 g/L MES.H2O. The pH was adjusted to 5.8 using 2 M NaOH or 1 M HCl before the addition of plant agar (8 g/L), and the medium was sterilized by autoclaving at 121 °C for 10 min | Freshly prepared | Induction of root emergence; seedling growth | [17] |
Rooting Medium (RM) | 1/2 MS medium containing vitamin, 10 g/L sucrose, 0.47 g/L MES.H2O, and 1 mg/L IBA. The pH was adjusted to 5.8 using 2 M NaOH or 1 M HCl before the addition of plant agar (8 g/L), and the medium was sterilized by autoclaving at 121 °C for 10 min | Freshly prepared | Induction of root emergence | [17] |
Root Regeneration Arabidopsis (RRA) | MS medium containing vitamin, 15 g/L sucrose, 0.47 g/L MES.H2O, 1 mg/L α-NAA, and 0.5 mg/L IBA. The pH was adjusted to 5.8 using 2 M NaOH or 1 M HCl before the addition of plant agar (8 g/L), and the medium was sterilized by autoclaving at 121 °C for 10 min | Freshly prepared | Induction of root emergence | [21] |