Expression Analysis Of Gapdh In Wheat Changwu 134 Under Three Kinds Of Abiotic Stree

GAPDH gene expression product plays an important role in catalyzing a carbohydrate metabolism which involves in plants defense reaction. Therefore, study on GAPDH gene expression and regulation is critical to increasing stress resistance and improve the yield of wheat. In this research, full length GAPDH gene sequences in drought-resistant wheat Changwu 134 and its 1071bp-length nucleotide sequences upstream were cloned, and the expression pattern of GAPDH gene was analyzed by real-time fluorescent quantitative PCR technology. To study the upstream promoter sequences responded to three kinds of abiotic stress transient expression of upstream promoters in Tobacco leaves, transfected with Agrobacterium strain (EHA105), was investigated. Furthermore, RNAi plants GAPDH gene recombinant expression vector was constructed and transformed to Arabidopsis to preliminary study the changwu143 GAPDH gene expression under abiotic stress to provide theoretical basis for Triticum application. Results are as follows:1. GAPDH gene of drought-resistant changwu143 wheat was unregulated under drought and low temperature stress while zhengyin 1which is much sensitive had no response to PEG600 treatment and low temperature. PEG6000 has the higtest induction effect. There is no significant change of GAPDH gene expression quantity in these two varieties under treatment of 250mM NaCl.2. Histochemical staining analysis indicated that GAPDH gene promoter was of high efficient and initiated the expression of GUS. The GAPDH promoter showed positive regulation pattern in drought, low temperature and ABA stresses. In addition, its activity was increased and was 3.43 times,2.13 times,1.75 times to the control respectively.3. RNAi expression vector was constructed successfully. Positive clone was identified by electrophoresis, digestion and DNA sequence analysis. Targeted gene was constructed with proper plant expression vectors.4. Eight strains are obtained by Kan positive and GUS positive treatment after the constructed RNAi transfected Arabidopsis.