Introduction Of OsNAC1、GAFP And ThpⅠ Genes Into Wheat By Gene Gun Bombardment

In recent years, China’s agriculture development very quickly, and the cultivation of wheat is the second place in all crops of China, The annual total production is basically about one billion tons. But at present China’s cultivated land decreased quickly, and the salinization of arable land is more and more serious, at the same time, high temperature, drought affect the high and stable production of wheat. So how to improve the resilience, frost resistance, resistance to pests and diseases of wheat has been the hot topic.The related stress resistance, disease and insect resistance genes from different organisms can be transfered by into wheat transgenic methods, to improve the resistance of wheat, resistance to diseases and insect pests. Currently, three commonly used methods of wheat transformation is:mieroprojectile bombardment, agrobacterium-mediated and pollen tube pathway. Because of no host restrictions, a wide range of receptors, high rate of plant regeneration and simple operation etc, The mieroprojectile bombardment become the main method of wheat transformation.There are a lot of research dedicated to finding a good target gene into wheat to improve resistance, disease and insect resistance and quality of wheat. In our research there are three target genes:OsNACl, GAFP, Thpl. OsNACl is a drought resistance and salt tolerant transcri-ption factor gene cloned from rice, can improve the drought resistance and salt tolerant of plant; GAFP is isolated and purified from Gastrodia, and the protein able to strongly inhibit fungal growth, can improve the disease resistance of wheat; The protein that Thpl encods which is an antifreeze protein isolated from Choristoneura fumigerana, it can improve the frost resistance of wheat.In this research, we used N13as receptor, put the OsNACl, GAFP and Thpl transferred to wheat by mieroprojectile bombardment. The main results are as follows:1. OsNACl, GAFP and ThpⅠ transferred to wheat by mieroprojectile bombardment(1) Put the OsNAC1, GAFP and ThpⅠ transferred into wheat by mieroprojectile bombardment, all the genes have gotten regenerated plants, the regeneration frequency was7.9%,7.3%and10.5%.(2) PCR analysis of To regeneration plants found that the OsNACl transcription factor obtained14positive plants, GAFP gene obtained1positive plant, ThpⅠ gene obtained4positive plants. The conversion rates were:0.82%,0.21%,0.85%.2. Detection and identification of the OsNAC1T1generation(1) We used PCR to detect the T1plants, ran out of seven positive bands, That the target gene has been transferred into wheat. Further evidence of target gene expression was increased in the T1transgenic plants came out by RT-PCR;(2) Using the250mmol/L NaCl to deal with the transgenic T1plants and the control group, The results showed that the Pro, SOD and POD of transgenic plants had significantly improved compared with N13.