Study On Protein Hereditary Property Transferring Soybean Total DNA Into Wheat By Low Energy Ion Beam

The method of low energy ion beam -mediated transformation was adopted to transfer the total DNA of soybean into yumai52 (It is the main planting breed of wheat in Henan province) in 1999. The offsprings of transformed wheat had been constantly analyzed and selected for four years. It was found that the protein content in seeds changed like the following trend: Tl had a wide range of protein content, an increase of 4.5% in average protein content than CK and many high-protein mutants; T2 had a larger range of protein content, an increase of 2.23% in average protein content than CK and few mutants, lots of high-protein individuals returned to the level of CK, but most of lines still left some high-protein mutants, the high-protein character separated obviously; T3 has an obvious increase of 7.84% in average protein content than CK, even among 58 lines the average protein content of 15 lines was higher than 17%, it began to take on the high-protein character and heredity in the unit of line; T4 has a more obvious increase of 15% in average protein content than CK, the protein distributed centrally and some high-protein lines were homologous, especially for 8 lines of T4 from 9063 to 9078 the average protein content were more than 16% and increased 24.2% than CK at least, and in the same line the protein of individual distributed centrally, only changed in a small range and took on the stable heredity.We set up the greenhouse generation-adding culture system to fit our breeding aims. The process was as fellows: The young embryos after pollination for 12~16d grew directly into seedlings, was moved into greenhouse after iarovization in 4-8 C for 25d, we controlled the temperature and humidity, made them blossom and pollination within 35~40d and then went on another circle of culture. Through thismethod we could finish the wheat's growth duration of one generation within about 70d. Culture the wheat continuously and we could finish five generations in a year. It cut the growth duration short obviously. Through mensurating the protein quality of seeds, we could forecast and guide the screening work in the farm, wash out revertant as soon as possible, and search into the stable high-protein mutants. The good result of T4's harvest in 2003 owed to this method.We respectively analyzed the gliadin in both the high-protein mutants of T4 and CK using A-PAGE. The band analysis of a , 3 and Y gliadin suggested that there was no difference of bands' addition or absence, but the bands' color of the high-protein wheat is deeper than CK's. It was the most different for w -gliadin. In the ?area, the number of protein bands increased obviously and the relative mobility of each protein band was also different. Analyzed the electrophoretic bands and we concluded that the heredity and stability of individuals in the 9065 line are better than in the 9106 line.We respectively analyzed the amino acid contents and the proportion of every kind of amino acid in both the high-protein mutants of T4 and CK. Judging by the proportion of essential amino acids, both high-protein lines is lower than CK, but in the whole, 17 kinds of amino acid contents (based on dry weight) increased obviously in the high-protein individuals of T4, especially Glu and Phe increased most. Furthermore Lys (the first restricted amino acid of wheat) respectively increased 48.10% and 37.00%.Analyzed the genome DNA of high-protein wheat of T4 and CK by 34 random primers using RAPD. 29 primers could produce stable and clear bands, while the bands produced by 18 primers were different in different degree. On the RAPD amplification illustration, the comparability between the high-protein wheat and CK was very high, and there were some addition, absence and different depth of the bands and so on. The individual's RAPD amplification from different high-protein lines took on the donor's (soybean) DNA bands that didn't exist in the receptor (wheat).The genome DNA of high-protein wheat of T4 and CK were analyzed by 64 couples of selective amplification primers' combin...