An improved method for rapid propagation of Phalaenopsis hybrids via culture of longitudinally bisected shoot tips.

L.K. Preetha, K.S. Shylaraj, P.C. Rohini

Abstract


A new procedure for in vitro propagation of orchids belonging to the genus Phalaenopsis was developed. In contrast to the commonly employed micropropagation methodsthat make use of dormant flower stalk buds or intact shoot tips, we have used longitudinally bisected shoot tips derived from 3 – 3.5 month old axenic plantlets of Phalaenopsis hybrids for protocorm-like-bodies (PLB) induction. This procedure in combination with the use of New Dogashima Medium for PLB proliferation resulted in large-scale production of plantlets. Culturing the bisected shoot tips in half strength solid Murashige and Skoog medium supplemented with Thidiazuron (TDZ) (4.54μM–9.08 μM) and 10% coconut water resulted in early PLB induction in 20-25 days and callus-free formation of 13.6 – 15.2 shoot buds in 8 weeks. Intact shoot tips took 6-8 weeks to PLB induction and proliferated to a mean of 5.46 - 7 shoot buds per shoot tip in 12 - 16 weeks in different TDZ concentrations. Continuous multiplication of PLBs and shoot buds was maintained in New Dogashima Medium devoid of hormones by bimonthly subcultivation. About 50,000 hardened plantlets could be produced from a single flowerstalkwith 10 dormant buds, within 2 years from initiation of axenic shoot tip cultures.This protocol can be used to produce genetically stable Phalaenopsis orchids on a commercial scale.


Keywords


Longitudinally bisected, Micropropagation, Phalaenopsis, Physiological age, Protocorm-likebody, Shoot tip

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References


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