Transformation Of CBF1 Gene Into Dianthus Chinese And Estimating Cold-resistance Of Transgenic Plant With CBF1 Gene

Dianthus chinese had been successfully transformed using a binary vector plasmidpBPCBFI which contained a chimeric nopaline synthase (NOS)promoter driven neomycinphosphotransferase (NPTⅡ) gene and a cauliflower mosaic virus 35S (CaMV35S)promoter driven CBFI (CRT/DRE binding factor 1)gene, and 4 transgenic lines wereobtained. Results of molecular analysis by PCR and physiological tests of cold resistance,confirmed the integration of CBF1 gene into the genome of these cultivars.The main research results were as follows:1. 28 transformed lines were detected by PCR analysis, and 4 of them showed thepositive band same as CBF1 gene. PCR positive Lines indicated the integration of CBF1gene into the genome of Dianthus chinese.2. The relative electric conductivity increased as S-curves with the temperaturedecreased. The relative electric conductivity of 4 trandgenic lines were less than CK ineach temperatre treatment. The relative electric conductivity of CK increased most at -20℃and was up to 505.03％.The transgenic line 27 increased least to 439.84％. Differencebetween CK and line 27 was much more significiant at -16℃and -20℃. Differencebetween CK and line 18 was significiant at -16℃. Difference between CK and line 12 wassignificiant at -16℃as same as line 18.3. Proline contents of control plant increased most and was up to 975.35％.Increasingof Proline contents of the transgenic lines was less than CK's.Their increasing were823.01％,747.59％,568.59％and 531.28％.Line 27 increased least.From -4℃to-16℃,increasing of each transgenic lines were 100％. Increasing of Proline contents of thetransgenic lines decreased to 50％.4. Malondialdehyde contents of each transgrnic lines and CK increased in differentlow temperature treatment. From 0℃to -20℃increasing of Malondialdehyde contents ofcontral line was 80.23％,12.14％,40.7％,39％and33.42％. Increasing of Malondialdehydecontents of each transgrnic lines was not so sharp as CK and just lasted about 20％.5.The activities of Superoxide Dismutase of each transgenic lines and CK increadfrom 0℃to -12℃in low temperature treatment.They reached top at -12℃and decreasedat -16℃. Difference between CK and each trnsgenic lines was much more significiant,butdifference between transgenic lines was not significiant.