Protocols for Micropropagation of Woody Trees and Fruits

MICROPROPAGATION OF ALBIZIA ODORATISSIMA
seedling 0.5 cm below the cotyledonary nodes to obtain cotyledonary nodes (1 cm
long) (Figure 2D). These two types of explants are suitable for developing protocols
for axillary shoot proliferation.
Cut the remaining hypocotyl region into small segments (1.5 cm long) (Figure
2E). This explant can be used for the development of protocol for adventitious shoot
organogenesis. Leaf nodes, cotyledonary nodes and hypocotyls obtained by using
the above procedure are shown in Figure 2F.
2.2.2. Collection and Surface Sterilization of Nodes from Two-year-old Saplings
Collect fresh, green and healthy top three branches of two-year-old saplings (Figure 3A).
Using surgical blade cut and remove the leaves attached to the nodes (Figure 3B,C). Cut
the stem portion of the branch 0.5 cm above and 0.5 cm below the nodes in order to
obtain leaf nodes (Figure 3D–F). Put the nodal explants in a non-sterile conical flask,
wash thoroughly under running tap water for 30 min. Immerse the explants in a detergent
solution (2 or 3 drops of tween-20 per 100 ml water) for 3 min and thoroughly
wash with non-sterile distilled water. Transfer the leaf nodes to a sterile conical flask.
Surface sterilize with 0.10% (w/v) mercuric chloride for 10 min. Under aseptic
condition wash thoroughly with sterile distilled water and inoculate immediately.
Figure 3. Procedure for the excision of leaf nodes from freshly collected branches of twoyear-old
saplings. A) Freshly collected young branches of two-year-old saplings. B–C)
Removal of leaves attached to the leaf nodes. D-E) Cutting the stem segment, separation of
leaf nodes. F) Leaf nodes excised from branches of two-year-old saplings.
2.3. Culture Medium
2.3.1. Basal Medium Components
The basal medium commonly used for Albizia tissue culture is MS (Murashige &
Skoog, 1962) with modifications to achieve stage dependent optimizations by particularly
manipulating the hormonal content. Table 1 lists the components of MS medium.
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