Protocols for Micropropagation of Woody Trees and Fruits

206
V. RAJESWARI AND K. PALIWAL
2.3.2. Medium Preparation
Prepare the basal medium including all additives except growth regulators as shown
in Table 1. Adjust medium to pH 5.7 with 1M NaOH and HCl and dispense in 150 × 25
mm culture tubes (15 ml per tube), 250 ml conical flasks (25 ml per flask). These
vessels are autoclaved for 1 min at 121°C and 1 × 105 Pa (1.1 Kg cm –2 ) and kept in a
cool place ready for culturing.
Table 1. Formulation of culture medium used for Albizia odoratissima micropropagation
based on MS salt augmented with culture stage-specific plant growth regulators.
Components Chemical formula Stock Medium
(g/L) (mg/L)
Macronutrients, 10 stock, use 100 ml per L medium
Ammonium nitrate NH4No3 16.5 1650
Potassium nitrate
Calcium chloride
KNO3
CaCl2
19 1900
.2H2O 4.4 440
Magnesium sulfate MgSO4 7H2O 3.7 370
Potassium orthophosphate KH2PO4 1.7
Micronutrients, 1000 stock use 1 ml per L medium
170
Boric acid
Cobalt chloride
H3BO3
COCl2
6.2 6.2
.6H2O
Cupric sulfate CuSO4
0.025 0.025
.5H2O
Manganese sulphate MnSO4
0.025 0.025
. H2O 16.9 16.9
Potassium iodide
Sodium molybdate
KI 0.83
Na2MoO4
0.83
. Zinc sulphate
2H2O 0.25
ZnSO4
0.25
. Iron sulfate
7H2O 8.6
Iron-EDTA, 100 , use 10 ml per L medium
FeSO4
8.6
. ×
×
×
7H2O 2.78 27.8
Ethylenediamine tetra
acetic acid di-sodium Na2EDTA 3.73 37.3
Myo-inositol
Vitamins, 100 ×
, use 10ml per L medium
10.0 100
Nicotinic acid 0.05 0.5
Pyridoxine hydrochloride 0.05 0.5
Thiamine hydrochloride 0.01 0.1
Glycine 0.2 2
Other additives
Sucrose 30000
Agar 8000
Plant growth regulators add according to the type of medium
Type of medium
MS-1 Half strength MS basal medium without plant growth regulators
MS-2 MS + 10 µMBAP and 1.5 µM NAA
MS-3 MS + 5 µM BAP and 1.5 µM NAA
MS-4 MS + 10 µM BAP and 10 µM 2-iP
MS-5 MS + 5 µM BAP and 10 µM kinetin
MS-6 MS + 7.5 µM BAP and 0.5 µM NAA
MS-7 MS + 25 µM IBA
MS-8 MS basal medium without plant growth regulators