Cloning of tungro resistance genes in rice through Mrna differential display
2001
Romero, G.O. | Solis, R.O. | Uera, R.B. (Philippine Rice Research Inst., Maligaya, Muñoz, Nueva Ecija (Philippines). Plant Breeding and Biotechnology Div.)
Tungro continues to be the most devastating rice disease, affecting many of the modern varieties including the widely popular IR64. This study aims to identify and clone the genes for resistance against tungro spherical virus (RTSV), the primary causal viral agent, to be used in the genetic engineering of tungro resistance (R) in the modern varieties. The author have identified potential R-related genes from examining the differentially displayed messages between two near-isogenic lines namely: tn1 (susceptible) and T1-11-8 (resistant). T1-11-8 contains the resistance genes) from an Indian landrace ARC11554. At 21 days after sowing, the R and S plants were inoculated with viruliferous green leafhopper for three days under mylar cage. At 20 days after inoculation, the R and S plants were phenotyped by ELISA method. At 21 days after inoculation, RNA was extracted and subsequent cDNA synthesis, PCR amplifications and gel electrophoresis were performed. Together with oligo (dT)G, four out of 20 arbitrary primers showed differential PCR products between the T1-11-8 (R) and the TN1 (S) lines on the agarose and polyacrylamide gels. The R-specific bands may represent R-related genes and are now under intensive cloning efforts.
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