Screening And Identification Of Susceptibility And Fertility Related Mutants From Transgenic Rice IRBB13(P_Xa13:GFP)

Rice Bacterial Blight is one of the most harmful bacterial disease in rice production,which widely distributed in the rice planting area and result in varying degrees of rice yield reduction. Many years of production practice show that cultivating disease-resistant variety is the most effective way to overcome bacterial blight. By constructing a mutant strategy is an effective way to study genes of unknown function. Using approach of EMS mutagensis method, we constructed a rice muation population with IRBB13(Pxa13:GFP) which carrying report gene green fluorescent protein(GFP) derived with dominant Xa13gene promoter. Then screening out the mutants with the following three periods:Scedling stage, using Stereo fluorescence microscope selected mutants in which GFP gene expression decline both in root and bud or non-expression; Adult stage, inoculating rice Bacterial Blight strains Philippine physiological race6PXO99,selecting recover susceptible phenotype mutants; Booting stage, selecting reduced spikelet fertility mutants and observed the GFP expression in pollen.Through microscopic examination,3,000M2generation lines were detected,14and9mutants lines out of3,000lines have been identified for both root and bud fluorescent declined and fluorescent normal only in root at seedling stage. In yield test,we inoculated and identified3,000M2generation lines during adult stage, a total of25lines have been primarily charactered to recover the susceptible to PXO99; At booting stage, a total of7lines was identified to reduced spikelet fertility mutants with anther green fluorescence disappeared, in which one mutant was found to recover susceptible too. These mutants obtained will lay the foundation for the cloning of rice bacterial blight resistance gene research and gene regulation of signaling networks pathway. In addition,we selected5pairs of uniform distributed SSR markers each chromosome in this experiment. Respectively carried out PCR amplification use rice varieties IRBB13(PXa13:GFP) and Long grain genomic DNA as template, Amplification products tested by capillary electrophoresis.Finally obtained21pairs of polymorphic molecular markers,out of which contained11pairs of codominant markers and10pairs of completely dominant markers.The obtain of these polymorphic molecular markers will lay the foundation for the localization of mutants genes.