Production And Heat-Tolerande Identification Of Transformed Lilum Longiflorum And Cyclamen Persicum With Mn-SOD Gene

The objective of this study was to research about Lilum longiflorum and Cyclamen persicum , Mn-SOD gene was incorporated into plants through Agrobacterium tumefaciens-mediated transformation in order to over-express SOD;After selection and confirmation , transformed lily and transformed cyclamen were acquired. And also researched on physiological changes of high temperature stress of transformed plantlets and non-transformed plantlets,expecting for evaluate its effect on the endurance to high temperature stress and environmental stress,pave the way for its breeding via genetic engineering.The main results were as follows:1 Abtained resistent lily plants by Agrobacterium-mediated genetic transformationSeveral factors that affected genetic transformation of Lilum longiflorum were examined,including the period of preculture and coculture,the time of infection of leaves,selection concentration of Kan,the concentration of Cef and so on. The result showed that the suiltable conditions of transformation contained 50mg/LKan,500mg/L Cef,2d preculture,3d coculture,10min infection,0.6 OD600. Thus a simple and efficient genetic transformation system was developed,and also abtained resistent lily plants.2 Screening out the best medium of adventitious bud regeneration through tuber and the ideal rooting medium of cyclamenWith the tubers as accepter,which come from the cyclamen seeds,the best medium of adventitious bud regeneration is MS+2.0mg/L6-BA+1.0mg/LNAA,and the treatment without light can enhance the regenerate rate. The ideal rooting medium of cyclamen is 1/2MS+1.0mg/LIBA or MS+1.5mg/LNAA.3 Confirmation of Mn-SOD gene31 resistant lily plants were confirmed by PCR assay,and 9 resistant plants showed a clear band corresponding to the relevant sequence of Mn-SOD gene,but the control plants were showed no band. Results of PCR proved preliminarily that the Mn-SOD gene had been transformed to Lilium longiflorum.5 PCR-positive resistant cyclamen plants (from Zhou Lian-xia)and 9 PCR-positive resistant lily plants were confirmed by Southern hybridization analysis,the results showed that 3 resistant cyclamen plants have a single band,this indicated that the Mn-SOD gene had integrated into cyclamen;and there is no band in resistant lily plants.4 Study on high temperature stress tolerance in Mn-SOD gene transformed cyclamen and lily plantletsPhysiological changes of high temperature stress at 36℃of transformed and control cyclamen and lily were measured. The experimental data showed that the level of SOD activity in transformed cyclamen was higher than control plantlets ; the membrane permeability and MDA content in transformed cyclamen rose more slowly than control plantlets;the protein content in transformed cyclamen was higher than control plantlets. These results indicated that the tolerance of the transformed plantlets to high temperature stress was higher than control plantlets. The level of SOD activity in transformed lily was also higher than control plantlets,the membrane permeability in transformed lily rose more slowly than control plantlets;the protein content in transformed lily was higher than control plantlets,but the MDA content had no discrepancy in evidence between transformed lily and control.