Transformation Of Gene AtAVP1, OsSIZ1 And FNR-FLD Into Wheat Immature Embryo By Microprojectile Bombardment And Agrobacterium Tumefaciens-Mediated Transformation

As a new method of crop breeding, genetic engineering technology has become the hotspot in gentic improvement study of wheat. The abiotic stress including salinization, drought, high temperature, low temperature and ion poisoninand is becoming more and more serious. With the harmful of abiotic factors for the growth and development of wheat, it had seriously threatted the wheat production in China. In order to achieve the aim of high and stable yield for wheat, it is great significative for cultivating a new wheat variety that it not only can resist the stress of abiotic but also can improve its own efficiency of photosynthesis. This study integrated the Arabidopsis thaliana vacuolar H+-pyrophosphatase(H+-PPase) gene(At AVP1), the ubiquitin E3 ligase gene(Os SIZ1),Ferredoxin NADP+ Reductase gene(FNR)from pea and Flavodoxin(FLD) gene from Anabaena to one plant expression vector by molecular cloning and then transformed it to the immature embryo callus. The receptor varieties include Zhengmai 9023, Huamai 2152 and Huamai 2566 and Huamai 2459. At last, we expect that new wheat materials with higher resistance and photosynthetic efficiency could be created. The results were as follows:1. We identified a total of 17 regeneration plants by the method of Agrobacterium-mediated genetic transformation, including one line Huamai 2459, 9 lines of Zhengmai 9023 and 7 lines of Huamai 2152. The regeneration plant derived from Huamai 2459 was negative, and each of Zhengmai 9023 and Huamai 2152 has only one positive line by PCR analysis.2. We also identified a total of 28 regeneration plants by biolistic method, including 15 lines of Huamai 2566, 4 lines of Bob White, 9 lines of Chinese Spring. However, we did not get positive transformant derived from Bob White, and each of Huamai 2566 and Chinese Spring has only one positive line by PCR analysis.3. Based on PCR analysis of 84 plants from Huamai 2152 T1 generation which were obtained by the Agrobacterium-mediated genetic transformation, none plants have been checked as positive lines. So the T0 generation of it may be false positive..4. The results also showed that 8 lines containing SIZ1 gene were positive from 20 T1 generation plants of Zhengmai 9023 by the method of Agrobacterium-mediated genetic transformation. two strains proved have AVP1 gene by PCR analysis,there did not detect positive FNR gene plant.