title: Establishment of an in vitro regeneration system for genetic transformation of selected  sugarcane genotypes.
abstract: A good  culture system provides considerable quantities of highly regenerable target  tissues. Embryogenic callus cultures are ideal for micro-projectile-mediated  transformation, because regenerable cells are not very stable. Effective  exploitation of genetic transformation requires good regeneration systems. We  selected three sugarcane genotypes for the establishment and optimization of  good in vitro regeneration systems, viz., S-2003-us-359, S-2006-sp-30,  and S-2003-us-165. Three callus induction media were investigated. These media  were composed of Murashige and Skoog (MS) medium salt plus 1, 2, and 3 mg/L  2,4-dichlorophenoxyacetic acid (2,4-D). Medium with 3 mg/L 2,4-D gave the  greatest mass of embryogenic calli. The calli produced on the three callus  induction media were transferred to 18 types of regeneration media (RM1-RM18).  They varied with respect to plant growth regulators and sucrose levels but the  basal medium was MS. Two levels of sucrose (30 and 40 g/L), three levels of  2,4-D (0.1, 0.25, 0.5 mg/L) and three levels of 6-benzylaminopurine (0, 0.25  and 0.5 mg/L) were studied in the regeneration media. The effects of callus age  on regeneration were evaluated by transferring the calli to regeneration media  after 15, 21, 28, and 35 days of culture. The 21-day-old callus of the genotype  S-2003-us-359 on RM3 yielded the largest number of plants and was selected as  the best for transformation. Six RAPD DNA primers were used to check genetic  stability; this medium did not affect the sugarcane genomes.
authors.affiliation: Centre of Agricultural Biochemistry and Biotechnology, University of Agriculture Faisalabad, Pakistan. siddraijazkhan@yahoo.com
authors.forename: S
authors.initials: S
authors.lastname: Ijaz
authors.forename: I A
authors.initials: IA
authors.lastname: Rana
authors.forename: I A
authors.initials: IA
authors.lastname: Khan
authors.forename: M
authors.initials: M
authors.lastname: Saleem
mesh_terms: D003470:Culture Media; D042822:Genomic Instability; D005838:Genotype; D018517:Plant Roots / Q000235:genetics / Q000254:growth & development; D018520:Plant Shoots / Q000235:genetics / Q000254:growth & development; D030821:Plants, Genetically Modified / Q000235:genetics / Q000254:growth & development; D012038:Regeneration; D031786:Saccharum / Q000235:genetics* / Q000254:growth & development; D014170:Transformation, Genetic
chemical_list: D003470:Culture Media