Protocols for Micropropagation of Woody Trees and Fruits

MICROPROPAGATION OF PLUM USING MATURE SEEDS 375
Figure 1. Explant excision. A) Seed without seed coat. B) Cotyledon with embryonic axis attached.
C) Embryonic axis dissected into segments (the epicotyl, the three slices of hypocotyl and the
radicle) as indicated.
Table 1. Composition of media used for plum micropropagation.
Component Final concentration (mg/l)
Induction medium Rooting medium
MgSO4·7H2O 370.0 185.0
MnSO 4·H2O 16.9
8.45
CaCl2·2H2O 444.0 222.0
NH4NO3 1650.0 825.0
KNO3 1900.0 950.0
KH2PO4 170.0 85.0
H3BO3 6.2 3.1
ZnSO4·7H2O 8.5 4.25
CuSO4·5H2O 0.025 0.0125
KI 0.88 0.44
CoCl2·6H2O 0.025 0.0125
Na2MoO4·2H2O 0.25 0.125
Fe(III)EDTA 0.0425 0.0213
Myo-Inositol 100 100.0
Thiamine·HCl 0.4 0.4
Nicotinic Acid 0.5 0.5
Pyridoxine·HCl 0.5 0.5
Indole-3-butyric Acid (IBA) 0.5 —
Naphthaleneacetic Acid (NAA) — 1.0
Kinetin — 0.002
Sucrose 25.0 10.0
Agar 7.0 7.0
Thidiazuron (TDZ) 1.66 —
Rooting medium (Table 1) consists of ½ strength MS salts (Murashige & Skoog,
1962), vitamins as above, 1.0 mg/L naphthaleneacetic acid (NAA), 0.002 mg/L
kinetin, 10 g/L sucrose, 7 g/L Bactoagar (Difco) with the pH adjusted to 5.9
(Gonzalez Padilla et al., 2003). After autoclaving, the medium may be poured into
25 × 100 mm Petri plates (35 ml) or into 10 cm Magenta containers (60 ml).
Murashige & Skoog basal salt mixture may be purchased from Phytotechnology Laboratories
(Shawnee Mission, KS, Catalogue # M524). Vitamins and hormones are prepared
as stock solutions (Table 2) to accurately add small quantities to the final media.