| Glyceraldehyde-3-phosphate dehydrogenase(GAPDH)is a key enzyme involved in the glycolysis and gluconeogenesis which plays a vital role in the energy supply for cells,and there are many researches revealed that the protein plays important roles in stress resistance in plants. In this study,two wheat cultivars,Changwu 134(CW134,drought-resistant) and Zhengyin 1(ZY1,drought-sensitive) were employed,and the RNA interference or the overexpression vector flanking open reading frame(ORF) of TaGAPDH8 was transformed into the callus induced by immature embryos via particle bombardment,respectively. After screening using hygromycin and identification by PCR,four down-regulated lines of CW134(CW134-3,CW134-6,CW134-12,and CW134-13) and eight up-regulated lines of ZY1(ZY1-1,ZY1-3,ZY1-4,ZY1-9,ZY1-10,ZY1-14,ZY1-15,and ZY1-17) were obtained. The growing characteristics of these T2 lines at milk-ripe stage in the field were measured to evaluate the influence of the exogenous gene on wheat plants. The lines that showed significant differences compared with CK lines were obtained. Subsequently,quantitative RT-PCR(qRT-PCR) analysis revealed that gene expression levels in the CW134 lines were 0.53,0.75,0.21,and 0.78-fold,and 3.02,1.22,2.15,1.36,4.02,1.87,1.48,and 1.97-fold in ZY1 lines,compared with that in the CK line,respectively. Our results indicated that we successfully obtained the T3 transgenic progeny with stable inheritance of the TaGAPDH8 gene,and these lines can be used for further study. |
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