Molecular characterization of transgenic rice (Oryza sativa L.) plants overexpressing mesophyll-specific carbonic anhydrase gene
2018
Mercado, M.A. | Bagunu, E. | Anonuevo, J.J. | Montecillo, F. | Reyes, J. | Balahadia, C.P. | Lin, H. | Xin, Y. | Coushoff, S. | Quick, W.P.
Carbonic anhydrase is an important enzyme in C4 photosynthesis, catalyzing the conversion of carbon dioxide to bicarbonate in the mesophyll cytosol. In this study, IR64 rice plants transformed with Maize carbonic anhydrase 1 gene driven with ZmPEPC promoter for mesophyll cytosol-specific expression and tagged with AcV5 epitope tag (ZmCA1-AcV5) were evaluated using different molecular techniques. Genomic PCR results showed 24 positive plants for the transformed gene. Immunoblotting showed protein expression in 16 out of 24 PCR positive plants. Varying copy number of genes was also observed in the Southern blot analysis. Cytological immunolocalization of cross-section of rice leaf samples showed strong protein expression of ZmCA1-AcV5 in cytosol of both mesophyll and bundle sheath cells of rice. T0 rice plants expressing the ZmCA1-AcV5 will be advanced for further screening and will later on be used as parent material for the gene-stacking of C4-photosynthesis related genes in rice.
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