Transforming Anti-TrxS Gene Into Wheat By Microprojectile Bombardment And Study On The Physiological Traits Of The Transgenic Plant

14 commercial wheat varieties from Huanghuai wheat district were used as materials to select the ideal acceptor in the transformation by microprojectile bombardment and to optimize the culture system of wheat transgenic regenerated plant. The result showed that four wheat varieties Yumail8-64, 00zhongl3, Zhengxin991 and Zhengxin992 had a high tissue culture ability; VBl(10mg/L) supplemented into MS media can improved the embryogenic callus development and increased the bud differentiation frequency; and 12h desiccation before differentiation culture can improved the bud differentiation frequency of calli. Based on this, we used the immature embryos (calli) of Yumai 18-64 as receptor to be bombarded with gold particles coated with the Anti-TrxS gene and bar selection gene, the two genes were co-transferred to Yumai 18-64 by bombardment. After callus induction regeneration on selective medium with 3mg/L Bialaphos, 42 resistant plantlets of TO generation were obtained. By PPT screening, there are 27 resistant wheat in the 42 regenerated plant, and the transformation frequency of bar gene is 1.27%. By PCR and PCR-Southern analysis in the 11 vigorous regenerated wheat from the 42 regenerated plant, 6 transgenic plants were identified with a transformation frequency of 2.11%. The co-transformation frequency of the two genes is 0.75%. For tracing the alien gene, the T1 and T3 plants were tested by PCR and PPT. The results demonstrated that the alien gene was integrated into the wheat genome. And the ratio of most Tl plant is 3:1 or 1:0, agreed with Mendel's Law of dominant gene with one locus, but that of T3 and some Tl plant is 1:1, which were not agreed with Mendel's Law. The reason is not so clear. Investigation also be carried out on the physiological traits which were relative to the Anti-TrxS gene. The results indicated that Trxh activity drop down significantlyin transgenic seeds comparing to CK, which led to a decrease in activity of a -amylase anddelay the germination of transgenic seeds. Anti-TrxS gene was responsible for these changes.