Study On The Transformation System Of Indica Rice With Antisense Lox-3 Gene

Rice is one of the most important grain crops in the world. Almost half of the world's population takes it as the main source of food and energy. So the stable increasing of rice yield ensures the world's food supply. However there is a large loss caused by aging deterioration, pests and mildew during rice storage. Therefore, carrying out a series of related researches on rice storable characteristics and breeding new storable rice varieties, both are very important.This study was designed to transform Cre/Lox-3 gene into indica restorer line"Hang No.1", expecting to select lipoxygenase low-expression lines from transgenic plants, making preparation for breeding new varieties with high storage resistance, and achieving the purpose of improving rice resistance to storage. Aiming at this goal, this study obtained the following conclusions:1. Analyzed the growth state of"Hang No.1"callus in six different media and their gain weight over different culture time, definited the optimum media for"Hang No.1".2. Plasmid pHG1PL, pHG4PL and pHG4P500L were transformed into indica restorer line"Hang No.1"respectively by Agrobacterium-mediated transformation, and 124 transgenic plants were obtained, the PCR results showed that the marker gene bar can be detected in 96 regenerated plants, preliminary indicating that the Cre/lox-3 gene had been integrated into indica restorer line"Hang No.1".3. Of altogether 96 PCR-positive transgenic plants, five ones were selected to perform Southern blot and dot blot analysis, and the result showed four transgenic plants detected the signal, indicating that the Cre/lox-3 gene had been integrated into their genomes; while one of them failed to detect any signal, indicating that the Cre/lox-3 gene had not been integrated into its genome, and the PCR result may be false positive.4. Selected 7 transgenic plants at random, the result of RT-PCR analysis showed that all of them with expression on mRNA lever, further indicated that the Cre/lox-3 gene had been integrated into indica restorer line"Hang No.1.5. Compared normal Agrobacterium-mediated transformation with early infection of agrobacterium, it was found that the transformation efficiency of the normal one was higher than the early infection, but it took much more time.6. Compared the microstructure of the root, stem and leaf from the normal and albino seedlings which were differentiated from the same clone transformed with Cre/lox-3 gene. It was found that there were some differences between the root, stem and leaf.