Effects of cytokinins on secondary somatic embryogenesis of selected clone Rayong 9 of Manihot esculenta Crantz for ethanol production

Abstract

In 2005, the Rayong-FCRC released a new high-yielding cassava cultivar designated Rayong 9 (R9) for ethanol production. However, the rate of distribution to farmers has been limited by the traditional vegetative propagation method of this crop which does not always satisfy the needs in planting material. The objective was to improve secondary somatic embryogenesis of the cassava clone Rayong 9 (R9) selected in Thailand by the Rayong Field Crops Research Center (Rayong-FCRC) for its suitability to produce ethanol. Fragments of cotyledon-stage somatic embryos were subcultured onto MS medium supplemented with the auxin 2,4-dichlorophenoxyacetic acid (2,4-D) at 4 mg/l in sight of inducing secondary somatic embryogenesis. Five different cytokinins, 6-benzylaminopurine (BAP), kinetin, zeatin, isopentenyladenine (2-iP) and adenine were added at 1 mg/l to the induction and maturation media to test their efficiency. Onto cytokinin-free media, 70 to 80% of explants produced embryoids, each explant giving 2 to 4 new embryoids within 7 weeks. The conversion rate of the embryoids into plantlets ranged from 11 to 26% depending on the type of cytokinin. With the exception of adenine, the other cytokinins inhibited the intensity of somatic embryogenesis, by 75% in the case of zeatin and 30% in the case of kinetin. Addition of adenine did not significantly improve the number of embryoids per explant. However, at 10, 20 and 40 mg/l adenine tended to improve the process relatively to embryoid sizes and plantlet survival rates in the greenhouse.

Key words: Adenine, biofuel, cassava, cotyledonary-stage, embryoids, icrocuttings, micropropagation.