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| Using the Phosphomannose Isomerase (PMI) Gene from Saccharomyces cerevisiae for Selection in Rice Transformation |
| WANG Tao, LIU Liang-yu, TANG Yong-yan, ZHANG Xiao-bo, ZHANG Mei-dong, ZHENG Yong-lian, ZHANG Fang-dong |
| National Key Laboratory of Crop Genetic Improvement, Ministry of Science and Technology/Huazhong Agricultural University, Wuhan 430070,P.R.China |
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摘要 The phosphomannose isomerase (PMI) gene from Saccharomyces cerevisiae acted as selectable marker and mannose acted as selective agent for the production of transgenic plants of rice (Oryza sativa L.) via Agrobacterium-mediated transformation. The concentration of mannose during the selection was stepwise increased, 5 g L-1 mannose combined with 15 g L-1 sucrose and 500 mg L-1 cefotaxime was used in the initial selection stage, then the concentration of mannose was increased to 11 g L-1, the highest transformation rate was 20.0%. The integration of PMI gene was confirmed by PCR, and the result of RT-PCR assay proved that the intron of PMI gene can be excised correctly during RNA splicing. β- Glucuronidase (GUS) activity analysis confirmed the expression of GUS gene. All those means the PMI gene from yeast can be used as a selectable marker in rice transformation.
Abstract The phosphomannose isomerase (PMI) gene from Saccharomyces cerevisiae acted as selectable marker and mannose acted as selective agent for the production of transgenic plants of rice (Oryza sativa L.) via Agrobacterium-mediated transformation. The concentration of mannose during the selection was stepwise increased, 5 g L-1 mannose combined with 15 g L-1 sucrose and 500 mg L-1 cefotaxime was used in the initial selection stage, then the concentration of mannose was increased to 11 g L-1, the highest transformation rate was 20.0%. The integration of PMI gene was confirmed by PCR, and the result of RT-PCR assay proved that the intron of PMI gene can be excised correctly during RNA splicing. β- Glucuronidase (GUS) activity analysis confirmed the expression of GUS gene. All those means the PMI gene from yeast can be used as a selectable marker in rice transformation.
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Received: 16 May 2011
Accepted:
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| Fund: This study was supported by the Genetically modified organisms breeding major projects，China（2011ZX08003-003） |
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Corresponding Authors:
Correspondence ZHANG Fang-dong, Tel: +86-27-87282689, E-mail: fdzhang@mail.hzau.edu.cn
E-mail: fdzhang@mail.hzau.edu.cn
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| About author: WANG Tao, E-mail: wt2915@163.com; |
Cite this article:
WANG Tao, LIU Liang-yu, TANG Yong-yan, ZHANG Xiao-bo, ZHANG Mei-dong, ZHENG Yong-lian, ZHANG Fang-dong.
2012.
Using the Phosphomannose Isomerase (PMI) Gene from Saccharomyces cerevisiae for Selection in Rice Transformation. Journal of Integrative Agriculture, 12(9): 1391-1398.
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