In vitro propagation of Cymbidium goeringii Reichenbach fil. through direct adventitious shoot regeneration.

Abstract

The influence of 2,4-dichlorophenoxyacetic acid (2,4-D), benzyladenine (BA), and thidiazuron (TDZ) on direct rhizome induction and shoot formation from rhizome explants of Cymbidium goeringii was explored. Rhizome segments obtained from in vitro seed cultures of C. goeringii were placed on Murashige and Skoog (MS) medium incorporated with 5, 10, 20, or 40µM 2,4-D and 1, 2, 4, or 8µM BA or TDZ alone or in combination with 20µM 2,4-D. The explants developed only rhizomes on MS medium with or without 2,4-D. The highest percent of rhizome formation (100%) was obtained on MS medium incorporated with 20μM of 2,4-D. The morphology and number of rhizomes varied with the level of 2,4-D in the medium. Direct adventitious shoot formation was achieved on medium incorporated with BA or TDZ. The adventitious shoots produced per explant significantly increased with the supplementation of 2,4-D to cytokinin-containing medium. The highest mean of 21.8±1.8 shoot buds per rhizome segment was obtained in medium fortified with 20μM 2,4-D and 2μM TDZ. The greatest percent of root induction (100%) and the mean of 5.3±1.1 roots per shoot were achieved on ½ MS medium incorporated with 2μM of α-naphthaleneacetic acid. About 97% of the in vitro-produced plantlets acclimatized in the greenhouse. An efficient in vitro propagation protocol was thus developed for C. goeringii using rhizome explants.