The Genetic Transformation Of Bt And Bt-pin Ⅱ Insect Resistance Gene In Ornamental Kale Via Agrobaterium Mediation

Ornamental kale(Brassica oleracea var.acephala)is an edible freezing tolerant excellent landscape plant,belonging to cabbge's horticulture varieties of Cruciferae Brassica.It is so prone to occur pest damage and freezing injury in its growing season that its production and ornamental value will be influenced.At present,there is no corresponding breeding resources.To endow it with pest insect resistance and increase the minus temperature tolerance further,there are a way will be operated through modern genetic engineering technology to create new resistant germplasm. However,breeding of ornamental kale is still in its infancy,and few international studies have been reported,lacking of detailed research data.Domestic improving their characters is still vacancy using genetic engineering technology.In this study,the plant expression vectors harboring insecticidal protein gene which were cloned were constructed and introduced into ornamental kale through genetic engineering technology,giving it new traits,to obtain good insect-resistant germplasm.The main results are as follows:1.A high efficient shoot regeneration system was established with different explants of eight genotypes representing the main phenotypes of ornamental kale.Two genotypes with 100%shoot induction rate on L6 or L3 medium respectively using cotyledon with petiole as explants were screened out,and the no.of shoots per explant were up to 25.The feasible rooting medium was MS+NAA 0.1 mg/L and the rooting rate reached to 100%.2.According to the studies on the relation between the infection time,the cultivation time,the sterilization of the explants,the ascertainment of ornamental kale the Basta screening pressure,the transgenic system of with Agrobacterium tumefaciens was constructed:infecting 3-5min;2d co-cultivation in dark;0.3-0.5 OD of bacterium..3.Three plant expression vectors pBAC171 and pBAC172,pBAC173,harboring a Bt...