Protocols for Micropropagation of Woody Trees and Fruits

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B. PINTOS ET AL.
Table 1. Formulation of culture medium used for doubled haploid micropropagation in
Quercus suber L. based on modified Sommer et al. (1975) macronutrients and Murashige &
Skoog (1962) micronutrients and augmented with culture stage-specific plant growth regulators.
Components
Chemical formula Stock
(g/L)
Macronutrients, 100× stock, use 10 ml per L medium
Medium
(mg/L)
Potassium chloride KCl 30 300
Potassium nitrate KNO3 100 1000
Calcium chloride-2H2O CaCl2·2H2O 15 150
Magnesium sulfate-7H2O MgSO4·7H2O 25 250
Ammonium sulfate (NH4)2·SO4 20 200
Sodium phosphate NaH2PO4·2H2O 12.950 129.50
Micronutrients, 100 × stock, use 10 ml per L medium
Potassium iodide KI 0.083 0.83
Boric acid H3BO3 0.62 6.2
Manganese sulfate-H2O MnSO4·H2O 1.69 16.9
Zinc sulfate-7H2O ZnSO4·7H2O 0.86 8.6
Sodium molybdate-2H2O Na2MoO4·2H2O 0.025 0.25
Cupric sulfate-5H2O CuSO4·5H2O 0.0025 0.025
Cobalt chloride-6H2O CoCl2·6H2O 0.0025 0.025
Iron –EDTA, 100 × stock, use 10 ml per L medium
Iron sulfate-7H2O FeSO4·7H2O 2.78 27.8
Ethylenediamine tetraacetic
acid disodium
Na2EDTA 3.72 37.2
Vitamins, 100 ×
stock, use 10 ml per L medium
Myo-Inositol 10 100
Nicotinic acid 0.05 0.5
Pyridoxine hydrochloride 0.05 0.5
Thiamine hydrochloride 0.01 0.1
Glycine 0.2 2
Ascorbic acid 0.2 2
Sucrose
Other additives
30000
Agar 8000
Plant growth regulators, glutamine and activated charcoal,
add according to culture stage
Final concentrations (mg/L)
Culture stage Glutamine Charcoal BAP IBA
Induction medium – 10000 – –
Proliferation medium 500 – – –
Maturation medium – 10000 0.05 –
Germination medium – – – 0.1