Wang Tao;Liu Liang-yu;Tang Yong-yan;Zhang Xiao-bo;Zhang Mei-dong;Zheng Yong-lian;Zhang Fang-dong

Huazhong Agr Univ, Natl Key Lab Crop Genet Improvement, Minist Sci & Technol, Wuhan 430070, Peoples R China

phosphomannose isomerase;selectable marker;Saccharomyces cerevisiae;transformation;rice

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. beta-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.