Protocols for Micropropagation of Woody Trees and Fruits

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B. PINTOS ET AL.
Table 2. Formulation of culture medium used for the induction of microspore embryogenesis
in olive. The culture medium is based on AT3 I medium (Höfer et al., 1999).
Components
Chemical formula Stock
(g/L)
Medium
(mg/L)
Macronutrients, 100 × stock, use 10 ml per L medium
Potassium nitrate KNO3 195 1950
Calcium chloride-2H2O CaCl2·2H2O 16.6 166
Magnesium sulfate-7H2O MgSO4·7H2O 18.5 185
Ammonium sulfate (NH4)2·SO4 27.7 277
Potassium bi-phosphate KH2PO4 40 400
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 MnSO4·4H2O 2.23 22.3
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 0.1 1
Pyridoxine hydrochloride 0.1 1
Thiamine hydrochloride 0.1 10
Maltose
Other additives
90000
MES 1950
Glutamine 1256
Table 3. The effect of temperature and duration of treatment on olive microspore division (%)
and development of multicellular structures per divided microspores (%) measured after 3
weeks of culture.
Treatments Microspore division
(%)
Multicellular structures/divided
microspores (%)
33°C 15 h 0.09 ± 0.00 0.09 ± 0.00
33°C 24 h 8.23 ± 1.24 15.23 ± 1.67
33°C 72 h 3.16 ± 2.00 2.54 ± 1.59
3°C 96 h 12.6 ± 1.15 32.44 ± 4.20