Somatic Embryogenesis and Genetic Transformation in Sugarcane Monzur Hossain and Rafiul Islam, Mohashweta Roy

An efficient protocol for somatic embryogenesis and subsequent plant regeneration was developed for sugarcane variety Isd-16 using leaf sheath explants of 3, 6 and 12-months-old field grown plants. Explants from 6-month-old plant showed the best response and produced highest percentage of calli on MS + 3.0 mg/l 2,4-D. L-proline 25mg/l significantly enhanced somatic embryogenesis. The embryos germinated well on half-strength MS and developed into plantlets. Somatic embryo derived plants under field condition showed considerable variation in morphological, agronomical and biochemical characters. For genetic transformation the calli were co-cultivated with A. tumefaciens strain LBA4404 harboring a binary plasmid pCAMBIA1303 containing hpt (hygromycin phosphotransferase) gene as a selectable marker and a ?-glucuronidase (gus) reporter gene in the T-DNA region. The transient expression of gus in hygromycin resistant calli and regenerated plants was confirmed by GUS flurometric assay. PCR... Это и многое другое вы найдете в книге Somatic Embryogenesis and Genetic Transformation in Sugarcane (Mohashweta Roy,Monzur Hossain and Rafiul Islam)