Molecular analysis of rice plants harboring a multi-functional T-DNA tagging system |
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Authors: | Yimian Ma Luo Liu Chengguang Zhu Changhui Sun Bo Xu Jun Fang Jiuyou Tang Anding Luo Shouyun Cao Gupo Li Qian Qian Yongbiao Xue Chengcai Chu |
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Institution: | [1]State Key Laboratory of Plant Genomics and National Plant Gene Research Center (Beijing), Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China; [2]Graduate University of the Chinese Academy of Sciences, Beijing 100039, China; [3]State Key Laboratory of Rice Biology China Institute of Rice Research, Chinese Academy of Agricultural Sciences, Hangzhou 310006, China; [4]Laboratory of Molecular and Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China |
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Abstract: | About 25,000 rice T-DNA insertional mutant lines were generated using the vector pCAS04 which has both promoter-trapping and activation-tagging function. Southern blot analysis revealed that about 40% of these mutants were single copy integration and the average T-DNA insertion number was 2.28. By extensive phenotyping in the field, quite a number of agronomically important mutants were obtained. Histochemical GUS assay with 4,310 primary mutants revealed that the GUS-staining frequency was higher than that of the previous reports in various tissues and especially high in flowers. The T-DNA flanking sequences of some mutants were isolated and the T-DNA insertion sites were mapped to the rice genome. The flanking sequence analysis demonstrated the different integration pattern of the right border and left border into rice genome. Compared with Arabidopsis and poplar, it is much varied in the T-DNA border junctions in rice. |
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Keywords: | rice mutant population promoter trap activation tag T-DNA integration |
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