Asian Science
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Abstract: An efficient method for in vitro propagation and cryopreservation
of Aerides odorata was established. Leaf segments were cultured on New
Dogashima (ND) mediums supplemented with various concentrations of Benzyladenine
(BA) (0-5 mg L-1) combined with Naphthaleneacetic Acid (NAA) (0-2
mg L-1). The optimal treatment for inducing Protocorm-like Bodies
(PLBs) from leaf segments was obtained from the combination of 1 or 3 mg L-1
BA and 0.5 or 1 mg L-1 NAA; whereas, the addition of BA or NAA alone
induced shoot and/or root initiation rather than PLB or callus formation. Shoots
rapidly developed on ND mediums containing 5 mg L-1 BA. Cryopreservation
of leaf segment-derived PLBs was successful using the encapsulation-dehydration
method. The maximum survival percentage of Cryopreserved (Cryp) PLBs was achieved
by encapsulating PLBs with 2% Na-alginate combined with 2 M glycerol and 0.4
M sucrose. The encapsulated PLBs were then precultured in 0.75 M sucrose for
24 h and dehydrated for 6 h before plunging into liquid nitrogen. Genetic stability
of Cryp PLBs after regrowth was assessed by flow cytometry. The findings showed
no different patterns of ploidy levels and morphology between Cryp and non-cryopreserved
(Ncryp) control plantlets.