Effect Of SNP And SA On The Expression Of Exogenous Genes And Methylation Level In Transgenic Brich

In this study, Different transgenic brich lines were selected, Then transgenic callus were treated by2mmol/L sodium nitroprusside (SNP) and50mg/L salicylic acid (SA). Physiological indicators and fluorescence quantitative PCR method were used to detect MET, DRM, BGT gene expression. Methylation sensitive type of different enzymes(HpaⅡ and MspⅠ) combined with RAPD molecular marker technology was applied to study transgenic brich epigenetic variation.Transgenic brich callus were treated by2mmol/L sodium nitroprusside and50mg/L salicylic acid. DRM and MET gene expression is higher in single-copy high expression of TP46than TP74, which were first increased and then decreased. In two clones, MET express activity is high in12h,24h and3d and the highest was approximately51.1times than control. DRM express activity is high in5d and the highest was approximately50times than control.The activity of SOD, POD, PAL and CAT were first increased and then decreased, While MDA content with trend of defense enzyme contrary. In TP46, the activity of defense enzyme is high in12h,3d and5d. Most notably the activity of SOD by SA was approximately2.34times than control in3d. In TP74, the activity of defense enzyme is high in5d and7d. Most notably the activity of SOD by SA was approximately8.22times than control in7d.Transgenic brich epigenetic variation were researched by methylation sensitive type of different enzymes (HpaⅡ and MspⅠ) combined with RAPD molecular marker technology. The results showed that TP46appeared209different bands, methylation sites from14.42%to16.54%and methylation level is highest in3d. TP74appeared257different bands, met-hylation sites from18.54%to20.81%and methylation level is highest in12h. Two clones are cytosine completely methylated sites in the inside mainly.