The Dynamic Study Of Resistance Regulatory Genes' In Plant Treated By Signaling Molecules Nitric Oxide And Salicylic Acid

Based on signaling transduction theory in plant cell,the dynamics of resistance signaling regulatory gene NtAC0,NPR1 and GST expression levels were studied in this paper by means of Gene Engineering,Molecular Biology etc.The sample leaves from the nicotiana tabacum cv xanthinc of 8~10 leaf period (with 8~10 leaves fully expanded)were divided into 3 groups.The1 and 2 group were treated by spraying 100ml 0.5 mmol/L NO donor sodium nitroprusside(SNP) and salicylic acid(SA)respectively . The 3 group was kept as the control group(CK)sprayed by Distilled Water.The samples of each group was tested at 0,12,24,36 hours to extract RNA and reverse transcript to cDNA. Using quantitative real time PCR and relative standard curve quantification to draw house keeping gene EF1-a and target genes NtACO,NPR1 and GST standard curve, The house keeping gene EF1-a and target genes NtACO,NPR1 and GST standards and samples were copied all together in 96 wells. Analysing the house keeping gene EF1-a and target genes NtACO,NPR1 and GST standard PCR Amp/Cycle Graph, standard Curve Graph, standard Melt Curve Graph, samples PCR Amp/Cycle Graph and samples Melt Curve Graph to discuss the relative expression levels of NtACO,NPR1 and GST . The results indicated that:1,During 12~36 h exogenous NO and SA dramatically inhibited the NtACO gene expression(P<0.01).Differently, the effect of NO was more than SA. At 12 h,24 h SNP treated samples the NtACO gene relative expression level decreased respectively 96.65 %,94.18%(P<0.01) compared to CK;But in SA treated samples the NtACO gene expression level decreased respectively 85.39 %,95.12 %(P<0.01) at 12 h,24 h compared to CK.2,During 12~36 h both NO and SA accelerated NPR1 gene expression, however, their trends were different.SNP treated samples the NPR1 gene expression first rised later declined, and the maximum expression level was 12 h,its relative expression increased 34.92 % (P<0.01)compared to 0 h ,the minimum expression level was 36 h ,its relative expression decreased 29.37 % (P<0.01)compared to 0 h;In SA treated samples the trend of the NPR1 gene expression level rised straight during the treated time(P<0.01).This indicated that the expression level increased along with the time extending,at12 h,24 h,36 h the NPR1gene relative expression increased respectively 43.14 %,91.45 %,214.33 %(P<0.01)compared to 0 h.3,During 12~36 h NO and SA had different regulatin to GST gene.NO inhibited GST gene expression(P<0.01),SA accelerated GST gene expression.At 12 h,24 h the SNP treated samples GST gene expression decreased respectively 46.36 %,73.37 %(P<0.01)compared to CK;SA treated samples the GST gene expression were all higher than CK, and the maximum expression level was 36 h,its expression increased 369.33 %(P<0.01) compared to 0 h,during 12~36 h GSTgene expression were higher than CK,at 12 h its expression increased 132.95 %(P<0.05),24 h increased 74.21 %,36 h increased 3645.90 %(P<0.01)compared to CK。...