Authors: Yuhya Wakasa Kenjirou Ozawa Fumio Takaiwa
Publish Date: 2012/07/28
Volume: 31, Issue: 11, Pages: 2075-2084
Abstract
A method for Agrobacteriummediated cotransformation of rice Oryza sativa L was developed using ricederived selection markers Two TDNAs were efficiently introduced into separate loci using selectable marker gene cassettes consisting of the mutated acetolactate synthase gene mALS under the control of the callusspecific promoter CSP CSPmALS and the ferredoxin nitrite reductase gene NiR under the control of its own promoter NiR PNiR The CSPmALS gene cassette confers sulfonylurea herbicide resistance to transgenic rice callus The NiR PNiR construct complements NiRdeficient mutant cultivars such as ‘Koshihikari’ which are defective in the regulation of nitrogen metabolism In the present study the CaMV35SGUS and CaMV35SGFP gene cassettes were cointroduced 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 coinoculated 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 cotransformation system The present results indicate that the generation of selectable antibiotic resistance marker genefree transgenic rice is possible using our ricederived selection marker cotransformation system
Keywords: