Genetic Transformation Of Single AtGolS2 Gene And SRK2C Gene In Populus×Euramericana Cv. 'Nanlin895'

Abiotic stresses such as drought,extreme temperature,salt injury which can affect the growth and quantity of forest trees sharply.In China,there are three million hectares are saline land,and arid area is increasing.So it is urgent to solve the abiotic stresses in forest research.At least seven GolS-related genes are found in the Arabidopsis genome.AtGolS2 is a member of GolS gene family in Arabidopsis thaliana.AtGolS2 were induced by drough and high-salinity stresses,but not by cold stress.SRK2C gene is a member of SnRK gene family.SnRK genes are capable of mediating signals initiated during drought,salt and cold stress,play important roles in transducing stress signals,increasing protein synthesization and regulating signal transduction.In this study,Populus×euramericana cv.'Nanlin895'was used as experiment material,adapting to Agrobacterium-mediated to introduce separately two resistance gene of AtGolS2 gene and SRK2C gene.The main conclusion of this experiment was as following:1.Determination of selective pressure of transformation:the selectable marker gene of AtGolS2 and SRK2C were both htp , so adopting Hyg as selective agent in transformation.Through contrast experiment,it is 2.5mg/L Hyg as selective pressure.2.Establishment of the efficient system of transformation:we studied several factors affecting Agrobacterium-mediated transformation,drawing the best transform condition: pre-culture time were 3 days, bacterial concentration OD600 0.8,infection time were 15～20 minute,coculture time were 4 days.3.Transgenic poplars were selected to survive on the culture medium with antibiotics,which includes Populus×euramericana cv.'Nanlin895'with single AtGolS2 gene and SRK2C gene.The molecular biological tests and salt stress experiment were carried out to verity these transgenic poplar.4.The two genes had been integrated into poplar genome and could express,which verified by PCR and RT-PCR.The result of salt treatment showed that salt-resistance ability of transgenic poplars with single AtGolS2 gene and SRK2C gene were enhanced.