Molecular and biochemical analysis of transgenic plants engineered for drought tolerance
2012
Liwanag, E. | Laluz, V., International Rice Research Inst. , DAPO 7777, Metro Manila 1301 (Philippines). Plant Breeding, Genetics and Biochemistry Div. | Natividad, M., International Rice Research Inst., DAPO Box 7777, Metro Manila (Philippines). Crop and Environmental Sciences Div. | Sallan-Gonzales, R., International Rice Research Inst., DAPO 7777, Metro Manila 1301 (Philippines). Plant Breeding, Genetics and Biochemistry Div. | Gaudin, A. | Henry, A., International Rice Research Inst., DAPO Box 7777, Metro Manila (Philippines). Crop and Environmental Sciences Div. | Slamet-Loedin, I., International Rice Research Inst., DAPO 7777, Metro Manila 1301 (Philippines). Plant Breeding, Genetics and Biochemistry Div.
Transgenic technology offers new prospects to increase rice drought tolerance by modulating expression of genes involved in drought response metabolic pathways. At IRRI Genetic Transformation Laboratory, genetic constructs harboring candidate genes for drought tolerance under constitutive and inducible promoters were introduced into IR64, a popular high yielding variety. A large number of independent transformants were obtained and screened for better yield under drought. Prior to screening, each event was genotyped and molecularly characterized. Gene integration and number of copies were first determined by PCR and Southern analysis. Hygromycin leaf painting and petri plate growing assays were developed to increase the efficiency of the genotyping and homozygosity determination based on mendelian segregation. Single copy, fertile T1 events were then selected and subjected to various drought environments in rain-out screen houses and contained field trials. To elucidate the correlation between the gene expression, phenotypic characteristic and certain physiology mechanisms, the authors conducted the real time and semi quantitative PCRs to qualify transgene expression levels and abscissic acid (ABA), osmolite, carbohydrate content analyses using ELISA and colorimetric technique. The flow and protocols of these biochemical and approaches are presented.
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