Anti-adversary Relevant DREB Gene Transferring Wheat And Functional Identification

Rd29A gene from Arabidopsis, encoding LEA-like protein, was induced by stress condition such as drought, high-salt and low-temperature. In this study , Rd29A promoter was used to construct gene expression vectors to control the expression of GUS gene. The vectors were transferred into wheat immature embryo calli by biolistic particle. The results of PCR, PCR-Southern and histochemical stain showed that GUS gene could express well induced by PEG treatment. Moreover, the result also indicated that the Rd29A promoter of Arabidopsis could be induced to regulate its downstream gene's expression in the stress-treated in wheat immature embryo calli. Then, the Rd29A promoter and constitutive Ubi promoter were applied to construct plasmid expression vectors with GH-DREB gene and GM-DREB gene respectively. They were pRdGH-Bar,pRdGM-Bar,pRdGH pRdGM,pRdGUS,pUbiGH,pUbiGM. All the constructs were proved by PCR and enzymatic digestion.In total, 6,000 wheat embryo calli were bombarded with the seven recombined plasmids and 220 transgenic plants were obtained.By using molecular marker-assisted selection, 59 positive transgenic plants were picked out. Then, 8 plants were further analyzed with RT-PCR, among which the specific fragment could be amplified from 4 plants. The result showed the desired genes had been transferred into wheat genome and got transcription. Mature embryos from the T1 seeds were cultured on the medium with 0.6% Nacl, 10% PEG for functional identification. The result demonstrated that the transgenic plants of LuMai 20 showed stronger resistantance to drought and high-salt than control, also stronger than those of YangMai 10.