Protocols for Micropropagation of Woody Trees and Fruits

356 B.K. BISWAS AND S.C. GUPTA
Figure 4. DNA finger printing of elite mother tree and in vitro raised plants. a) Mother plant
growing as an avenue tree on Probyn Road, Delhi. b) In vitro raised plants after 1 month of
transfer to soil in greenhouse. c,d) Evaluation of genetic fidelity of micropropagated neem
plants through RAPD analysis. RAPD profiles generated using primers OPA-13 (c) and
OPC-14 (d).
3. CONCLUSIONS
This chapter describes biochemical analysis of azadirachtin to select a 35-year-old
elite tree of Azadirachta indica A. Juss., its clonal propagation protocol and
assessment of genetic fidelity of the regenerants by RAPD analysis. The neem tree,
of Indian origin, is highly valued worldwide as a source of medicine and a potent
biopesticide, whose active principle is azadirachtin – a triterpenoid. The selection of
the elite neem tree, based on azadirachtin content determination in its seed kernels
using TLC and HPLC, has increased the significance of this study.
The highest azadirachtin content (0.4%) has been reported from Myanmar while
presently a neem tree with 0.746% in its seed kernels has been identified which is
the highest ever known record in the world.
Most of the published protocols on neem involve the juvenile tissues while the
success with physiologically mature explants has been limited. However, it is essential