AN EFFICIENT, SHORT AND COST-EFFECTIVE REGENERATION SYSTEM FOR TRANSFORMATION STUDIES OF SUGARCANE (SACCHARUM OFFICINARUM L.)

Abstract

Sugarcane genetic transformation efforts are seriously hampered by the lack of an efficient and reproducible regeneration system. An efficient, short and cost-effective regeneration system, through direct embryogenesis, was developed for local cultivars and elite lines of sugarcane. Using 1-2 mm thick meristematic young leaf whirls, direct embryogenesis was achieved in Murashige & Skoog (MS) medium supplemented with 2, 4-dichlorophenoxyacetic acid (2, 4-D) under cool white fluorescent light for 16 hour/day at 25 ± 2 °C within three weeks. Of the various concentrations of 2, 4-D tested, 3 mg/L induced the highest frequency of embryogenic callus (60 %). The embryos germinated in the fourth week on the same medium. Shoot proliferation and multiplication was carried out in liquid MS medium containing benzyl aminopurine (BAP) at a concentration of 1 mg/L. The improved regeneration system will particularly be useful in our ongoing genetic transformation studies.