Using the Phosphomannose Isomerase (PMI) Gene from Saccharomyces cerevisiae for Selection in Rice Transformation

doi:10.1016/S1671-2927(00)8670

Journal of Integrative Agriculture ›› 2012, Vol. 12 ›› Issue (9): 1391-1398.DOI: 10.1016/S1671-2927(00)8670

• 论文 • 下一篇

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

收稿日期:2011-05-16 出版日期:2012-09-01 发布日期:2012-09-09
通讯作者: Correspondence ZHANG Fang-dong, Tel: +86-27-87282689, E-mail: fdzhang@mail.hzau.edu.cn
作者简介:WANG Tao, E-mail: wt2915@163.com;
基金资助:
This study was supported by the Genetically modified organisms breeding major projects，China（2011ZX08003-003）

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

Received:2011-05-16 Online:2012-09-01 Published:2012-09-09
Contact: Correspondence ZHANG Fang-dong, Tel: +86-27-87282689, E-mail: fdzhang@mail.hzau.edu.cn
About author:WANG Tao, E-mail: wt2915@163.com;
Supported by:
This study was supported by the Genetically modified organisms breeding major projects，China（2011ZX08003-003）

摘要/Abstract

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.

关键词: phosphomannose isomerase, selectable marker, Saccharomyces cerevisiae, transformation, rice

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.

Key words: phosphomannose isomerase, selectable marker, Saccharomyces cerevisiae, transformation, rice

WANG Tao, LIU Liang-yu, TANG Yong-yan, ZHANG Xiao-bo, ZHANG Mei-dong, ZHENG Yong-lian, ZHANG Fang-dong. Using the Phosphomannose Isomerase (PMI) Gene from Saccharomyces cerevisiae for Selection in Rice Transformation[J]. Journal of Integrative Agriculture, 2012, 12(9): 1391-1398.

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Using the Phosphomannose Isomerase (PMI) Gene from Saccharomyces cerevisiae for Selection in Rice Transformation

Using the Phosphomannose Isomerase (PMI) Gene from Saccharomyces cerevisiae for Selection in Rice Transformation

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