Establishment Of Transformation System In High Oleic Soybean Via Agrobacterium Tumefaciens

The genetic transformation of soybean is the main barrier which is defines to theresearch of transgenic soybean. Although this work has been studied for many years, andseveral transformation systems were established, none of their improving is of the essence.For the sake of better and faster development of genetically engineered soybean, it ismeaningful to establish a high efficiency transformation method.A soybean species named Jilin 35, the main soybean cultivars in Jilin province, was usedin this study. The gmFAD2-1 gene which is seed-specifically-expressed was transferred intosoybean mediated by Agrobacterium tumefaciens EHA105, based on the dsRNA-mediatedgene silencing technology. According to recent research advance of soybean transformation,three genetic transformation systems were established and optimized: cotyledon nodestransformation system refers to the method of Iowa University of USA, embryonic tiptransformation system refers to Liu's newest report;shoot apex transformation systemrefers to a maize shoot apex transformation method of Zhang. In this study, we investigatedthe efficiency and advantages of these three transformation systems, furthermore, sometransgenic plants were obtained.Our experiment results showed that the embryonic tips transformation system had higherregeneration frequency, more sensitive to selected agent (Kanamycin), shorter regenerationcycle, and easier to get resistant plants compared to the cotyledon nodes transformationsystem. The embryonic tips transformation has a lot of advantage over cotyledon nodestransformation system, and it will be one of best choice of soybean genetic transformation.Contrast with the other two systems the shoot apex transformation system is easier tooperate and get resistant plants, but the one of most disadvantage is it needs a great deal ofselection work. The soybean genetic transformation via Agrobacterium tumefaciens was improving andinnovative, and a set of soybean transformation systems were optimized and established inthis research. Our experimental work is mainly aim at to select a transgenic high oleicsoybean lines. Meanwhile, we hope to establish a set of success soybean transformationalsystem to provide groundwork for the research of transgenic soybean and transformation.