TISSUE CULTURE TECHNIQUES FOR TREE SPECIES
Media for Culture
Media is one of the important component of tissue culture. The media is the source of macronutrients, micronutrients, vitamins, growth regulators, carbon etc., which are needed by cell for their growth and differentiation. The various media composition that are used commonly in tissue culture are presented (Table 1)
Table 1. Standard Basal Media Formulations Commonly used in Plant Tissue Culture
| 1. MS Basal Medium (Murashige & Skoog, 1962) |
| Compenents |
Quantity (mgl-1) |
| Macronutrients |
| NH4NO3 |
1650.0 |
| KNO3 |
1900.0 |
| CaCI22H2O |
440.0 |
| MgSO4.7H2O |
370.0 |
| KH2PO4 |
170.0 |
| Micronutrients |
| KI |
0.83 |
| H2BO3 |
6.20 |
| MnSO4.4H2O |
22.30 |
| ZnSO47H2O |
8.60 |
| Na2MoO4.2H2O |
0.25 |
| CuSO45H2O |
0.025 |
| CoCI2.6H2O |
0.025 |
| Na2EDTA |
37.30 |
| FeSO4.7H2O |
27.80 |
| Vitamins and other supplements |
| Inositol |
100.0 |
| Glycine |
2.0 |
| Thiamine |
0.1 |
| Pyridoxine HCI |
0.5 |
| Nicotinic acid |
0.5 |
| 2. B5 Basal Medium (Gamborg et al., 1968) |
| Macronutrients |
| KNO3 |
3000.0 |
| CaCI2.2H2O |
150.0 |
| MgSO4.7H2O |
500.0 |
| (NH4)2SO4 |
134.0 |
| NaH2PO4H2O |
150.0 |
| Micronutrients |
| Kl |
0.75 |
| H3BO3 |
3.00 |
| MnSO4.4H2O |
10.00 |
| ZnSO47H2O |
2.00 |
| Na2MoO4.2H2O |
0.25 |
| CuSO45H2O |
0.025 |
| CoCI2.6H2O |
0.025 |
| Na2EDTA |
37.30 |
| FeSO4.7H2O |
27.80 |
| Vitamins and other supplements |
| Inositol |
100.00 |
| Thiamine HCI |
10.0 |
| Pyridoxine HCI |
1.0 |
| Nicotinic acid |
1.0 |
| 3. White’s Basal Medium (White. 1963) |
| Macronutrients |
| KNO3 |
80.0 |
| MgSO4.7H2O |
720.0 |
| NaH2PO4. |
16.5 |
| Ca(NO3)2.4H2O |
300.0 |
| Micronutrients |
| Kl |
0.75 |
| H3BO3 |
1.50 |
| MnSO4 |
7.00 |
| ZnSO4.7H2O |
2.60 |
| Vitamins and other supplements |
| Glycine |
3.0 |
| Thiamine HCl |
0.1 |
| Pyridoxine HCl |
0.1 |
| Nicotinic acid |
0.5 |
| Calcium pantathenate |
1.0 |
| Cysteine HCl |
1.0 |
| Woody Plant Medium(Lloyd and McCown, 1980) |
| Macronutrients |
| NH4NO3 |
400.0 |
| KH2PO4 |
170.0 |
| MgSO47H2O |
370.O |
| K2SO4 |
980.0 |
| Ca(NO3)24H2O |
556.0 |
| CaCl22H2O |
96.00 |
| Micronutrients |
| H3BO3 |
6.20 |
| CuSO4.5H2O |
0.25 |
| MnSO4.4H2O |
22.30 |
| ZnSO4.7H2O |
8.80 |
| Na2MoO4.2H2O |
0.25 |
| Iron EDTA |
| Na2EDTA |
37.30 |
| FeSO47H2O |
27.80 |
| Organic nutrient |
| Pyridoxine |
0.50 |
| Myoinositol |
100.00 |
| Glycine |
2.00 |
| Thiamine HCl |
0.10 |
| Nicotinic acid |
0.50 |
|
