Establishment Of Micropropagation Protocol For Banana (Musa acuminata x M. balbisiana) Cv. Pisang Awak (Abb Genome) Via Temporary Immersion System
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Date
2011-02
Authors
Au, Vun Hui
Journal Title
Journal ISSN
Volume Title
Publisher
Universiti Sains Malaysia
Abstract
Two surface-sterilization protocols were established to obtain aseptic shoot
explants of Pisang Awak for micropropagation purpose. Aseptic explants could be
established from one day old suckers by using one-stage surface-sterilization method
with 20% (v/v) Clorox® for 20 minutes. Two-stage surface-sterilization technique
could be used for more than one day old suckers. This protocol involved surfacesterilization
with 20% (v/v) Clorox® for 20 minutes at the first stage followed by
10% (v/v) Clorox® for 10 minutes at the second stage, then inoculated onto solid MS
medium supplemented with 150 mg/L Kanamycin for seven days. Shoot proliferation
was achieved by culturing the aseptic half shoot explants onto solid or liquid MS
medium supplemented with 5.0 mg/L BA to induce multiple shoots formation. The
multiple shoots were separated individually and best rooting could be achieved using
solid basic MS medium. Plantlets with a minimum height of 2.0 cm and well
developed root system could be acclimatized very well in the greenhouse.
Application of 0.75 g/L NPK fertilizer (15:15:15) during secondary hardening
process enhanced the growth of Pisang Awak plantlets. A mass propagation protocol
using a modified temporary immersion system (TIS) was established. Optimum
number of plantlets could be achieved in TIS by inoculating eight half shoots in 100
mL MS medium supplemented with 5.0 mg/L BA and 30 g/L sucrose at pH 5.7
immersed for 10 minutes every 24 hours under continuous light condition. The
optimized TIS protocol for the mass production of Pisang Awak plantlets was also
suitable for mass production of Pisang Raja plantlets. TIS was proven to be more
efficient than gelled medium and shake flask system for shoot proliferation of Pisang
Awak. Rooted micro-shoots established from TIS could be acclimatized directly to
greenhouse condition with 93% survival percentage. While micro-shoots without
root established from gelled medium and TIS needed to undergo rooting in basic MS
medium and 89 - 92% of the plantlets survived after acclimatized in the greenhouse.
Description
Keywords
Two surface-sterilization protocols were established to obtain , aseptic shoot explants of Pisang Awak for micropropagation purpose