title: Agrobacterium-mediated co-transformation of rice using two selectable marker genes derived from rice genome components.
abstract: A method for Agrobacterium-mediated co-transformation of rice (Oryza sativa L.) was developed using rice-derived selection markers. Two T-DNAs were efficiently introduced into separate loci using selectable marker gene cassettes consisting of the mutated acetolactate synthase gene (mALS) under the control of the callus-specific promoter (CSP) (CSP:mALS) and the ferredoxin nitrite reductase gene (NiR) under the control of its own promoter (NiR P:NiR). The CSP:mALS gene cassette confers sulfonylurea herbicide resistance to transgenic rice callus. The NiR P:NiR construct complements NiR-deficient mutant cultivars such as 'Koshihikari', which are defective in the regulation of nitrogen metabolism. In the present study, the CaMV35S:GUS and CaMV35S:GFP gene cassettes were co-introduced into the 'Koshihikari' genome using our system. Approximately 5-10 independent transgenic lines expressing both the GUS and GFP reporters were obtained from 100 Agrobacterium co-inoculated calli. Furthermore, transgenic 'Koshihikari' rice lines with reduced content of two major seed allergen proteins, the 33 and 14-16 kDa allergens, were generated by this co-transformation system. The present results indicate that the generation of selectable antibiotic resistance marker gene-free transgenic rice is possible using our rice-derived selection marker co-transformation system.                 Key message An improved rice transformation method was developed based on Agrobacterium-mediated co-transformation using two rice genome-derived selectable marker gene cassettes.
authors: Functional Transgenic Crops Research Unit, Genetically Modified Organism Research Center, National Institute of Agrobiological Sciences, Kannondai 2-1-2, Tsukuba, Ibaraki, 305-8602, Japan. ywakasa@nias.affrc.go.jp
authors: Yuhya
authors: Y
authors: Wakasa
authors: Kenjirou
authors: K
authors: Ozawa
authors: Fumio
authors: F
authors: Takaiwa
mesh_terms: D000095:Acetolactate Synthase / Q000235:genetics; D060054:Agrobacterium / Q000502:physiology*; D000485:Allergens / Q000235:genetics; D004269:DNA, Bacterial; D050900:Ferredoxin-Nitrite Reductase / Q000235:genetics; D017930:Genes, Reporter; D005819:Genetic Markers; D053209:Herbicide Resistance; D009154:Mutation; D012275:Oryza / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012038:Regeneration; D012639:Seeds / Q000201:enzymology / Q000235:genetics; D013453:Sulfonylurea Compounds / Q000494:pharmacology; D014170:Transformation, Genetic
chemical_list: D000485:Allergens; D004269:DNA, Bacterial; D005819:Genetic Markers; D010940:Plant Proteins; D013453:Sulfonylurea Compounds; C036483:T-DNA; D050900:Ferredoxin-Nitrite Reductase; D000095:Acetolactate Synthase