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GENOMICS, MOLECULAR GENETICS & BIOTECHNOLOGY

Engineering Higher Yield and Herbicide Resistance in Rice by Agrobacterium-Mediated Multiple Gene Transformation

^a National Lab. of Plant Molecular Genetics, Inst. of Plant Physiology and Ecology, Shanghai Inst. for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China
^b Agrobiotech Research Center, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China
^c Crop Breeding and Cultivation Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China

^* Corresponding author (jqhuang{at}iris.sipp.ac.cn)

The Vitreoscilla hemoglobin gene (VHb), trans-zeatin secretion gene (tzs), and the modified 5-enolpyruvylshikimate-3-phosphate synthase gene (EPSPS), as linked expression cassettes, were simultaneously introduced into immature embryos of the rice (Oryza sativa L.) cultivars Xiushui-11, Qiufeng, Youfeng, and Hanfeng by Agrobacterium tumefaciens. A total of 1153 transgenic lines composed of 4222 plants were obtained through selection for hygromycin (hyg) B resistance. Genomic polymerase chain reaction (PCR), southern and northern blotting analyses, and other relative tests showed that all transgenes had been integrated into the rice genome and expressed effectively. Approximately 90.2% of the transgenic lines harbored all the transgenes. Expression analysis revealed that all transgenes coexpressed stably in transgenic plants, and the frequency of coexpression was about 85%. Statistically significant increases were observed in plant height, panicle length, total grains per panicle, and filled grains per panicle in transgenic plant lines compared with the control. Our study demonstrates a possible way to introduce different transgenes as linked expression cassettes within a single vector into the plant genome. Moreover, this transgenic approach has great potential in developing new rice cultivars with increased productivity and enhanced tolerance to the herbicide glyphosate.

Abbreviations: AS, acetosyringone • DIG, digoxigenin • EPSPS, 5-enolpyruvylshikimate-3-phosphate synthase • EPSPS, modified 5-enolpyruvylshikimate-3-phosphate synthase gene • GUS, ß-D-glucuronidase • hpt, hygromycin phosphotransferase gene • hyg, hygromycin • ipt, isopentanyl transferase gene • MAR, matrix attachment region • PCR, polymerase chain reaction • RBCS, small subunit of ribulose-bisphosphate carboxylase • SAR, scaffold attachment region • SCC, standard saline citrate • tzs, trans-zeatin secretion gene • VHb, Vitreoscilla hemoglobin gene

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Crop Science 2004 44: 1889-1892. [Full Text]  

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