PJB-2009-68
AN EFFICIENT, SHORT AND COST-EFFECTIVE REGENERATION SYSTEM FOR TRANSFORMATION STUDIES OF SUGARCANE (SACCHARUM OFFICINARUM L.)
AZAD HUSSAIN SHAH*, NAEEM RASHID, M. SALEEM HAIDER, FAIZA SALEEM, M. TAHIR AND JAVED IQBAL
DOI:
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.
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