Construction Of Binary Expression Vectors Containing Lyz./Chi. Genes And Organogenesis Regenerating System Of Licorice

Glycyrrhiza uralensis Fisch is Glycyrrhiza leguminous perennial herbs, which are widely used in cosmetics, food, tobacco, pharmaceutical and other industries. Meanwhile, Glycyrrhiza uralensis Fisch is an important Sand-fixing plants of western desert and semi-desert areas of china.In recent years, due to Glycyrrhiza uralensis Fisch diseases and People devastating excavation, wild Glycyrrhiza uralensis Fisch resources almost extinguished. Therefore, the cultivation of high quality disease-resistant varieties of Glycyrrhiza uralensis Fisch for its production is of great significance.In recent years, great achievements have been obtained in genetically engineering plants resistant to fungal and bacteria disease. Several genes for resistance to fungal and bacteria disease have been cloned and demonstrated, leading to many disease resistant plants with stable heredity. Plant gene engineering techniques offer new and efficient methods for breeding cultivars resistant to disease.In this study, we have finished the two aspects of work, we selected the Glycyrrhiza uralensis Fisch hypocotyls as explants, and study a variety of physiological, physical,chemicalby change impact on the shape of callus induced by the hypocotyl by controlling hormone levels, sucrose concentration, temperature, light, active carbon levels.then we use the seed embro as explants,induced callus,and determined the best medium,eventually we got dividing seedings. (2)This study successfully constructed a binary-expression vectors containing Lysozyme and Chitinase, the test results of PCR reaction and enzyme identification proved Chi gene and Lyz gene were positively inserted into PBI-Lyz-Chi and target gene linked end-to-end . After sequencing,the PBI-Lyz-Chi vector was used transformed tumefacience LBA4404 to abtain the engineering Agrobacterium strains harboring the target genes.All strains were tested by PCR reaction and enzyme identification. (3)Transformation of Glycyrrhiza uralensis Fisch mediated by A.tumefacience were performed.some data have been abtained, it would provide basal data for next work.