title: Agrobacterium-mediated genetic transformation of commercially elite rice restorer line using nptII gene as a plant selection marker.
abstract: Transformation of commercially important indica cultivars remains challenging for the scientific community even though Agrobacterium-mediated transformation protocols for a few indica rice lines have been well established. We report successful transformation of a commercially important restorer line JK1044R of indica rice hybrid JKRH 401. While following existing protocol, we optimized several parameters for callusing, regeneration and genetic transformation of JK1044R. Calli generated from the rice scutellum tissue were used for transformation by Agrobacterium harboring pCAMBIA2201. A novel two tire selection scheme comprising of Geneticin (G418) and Paramomycin were deployed for selection of transgenic calli as well as regenerated plantlets that expressed neomycin phosphotransferase-II gene encoded by the vector. One specific combination of G418 (30 mg l(-1)) and Paramomycin (70 mg l(-1)) was very effective for calli selection. Transformed and selected calli were detected by monitoring the expression of the reporter gene uidA (GUS). Regenerated plantlets were confirmed through PCR analysis of nptII and gus genes specific primers as well as dot blot using gus gene specific as probe.
authors.affiliation: Department of Biotechnology, Jawaharlal Nehru Technological University (JNTU), Hyderabad, Telangana 500072 India ; Biotechnology Division, J.K Agri. Genetics Pvt. Ltd., Hyderabad, Telangana 500016 India.
authors.forename: M
authors.initials: M
authors.lastname: Chakraborty
authors.affiliation: Biotechnology Division, J.K Agri. Genetics Pvt. Ltd., Hyderabad, Telangana 500016 India.
authors.forename: P
authors.initials: P
authors.lastname: Sairam Reddy
authors.affiliation: Department of Biotechnology, Jawaharlal Nehru Technological University (JNTU), Hyderabad, Telangana 500072 India.
authors.forename: M
authors.initials: M
authors.lastname: Laxmi Narasu
authors.affiliation: Biotechnology Division, J.K Agri. Genetics Pvt. Ltd., Hyderabad, Telangana 500016 India ; Jacob School of Biotechnology & Bioengineering, Sam Higginbottom Institute of Agriculture, Technology & Sciences (Formerly Allahabad Agricultural Institute), Deemed University, Allahabad, 211007 Uttar Pradesh India.
authors.forename: Gaurav
authors.initials: G
authors.lastname: Krishna
authors.affiliation: Biotechnology Division, J.K Agri. Genetics Pvt. Ltd., Hyderabad, Telangana 500016 India ; Bayer CropScience-Seeds, Bayer (South East Asia) Pte Ltd, Singapore, Singapore.
authors.forename: Debashis
authors.initials: D
authors.lastname: Rana
keywords: Agrobacterium tumefaciens; Genetic transformation; embryonic callus; Geneticin (G418); NptII selection; Paramomycin; Restorer line; Rice (Oryza sativa); UidA (GUS)