| From 2007 to 2009,with the methods of pot experiment, the effects of N forms on the expression and Subunit composition of GS isozymes in different organs during different phase of yumai34 were studied, and the GS isozymes In Wheat were Separated and sublimated, the subunits composition of GS protein were Characterized and evaluated.the main results were as follows:By the pot experiment in field, simulating field, the effects of N forms on the expression of GS in leaves of yumai34 were studied. The results indicated: the expression of GS isozymes in leaves of the seeding stage was little affected by N forms and genotypes, but affected greatly by different growth stages. There are three types of GS isozymes in all phases of seeding stage, but compared the other three phases, the GS activity was lower in the stage of one leave and one heart. Compared to the expression of GS isozymes in the seeding stage, a new isozyme GSX2 was found in the leaves of anthesis, the ammonium has certain function in inducing the appearance of GSX2. It appears in anthesis and 7DAF only. There are also four types of isozymes expressed in the stem in the same stage, the GS1 isozyme occupy the dominant position, and only GS1 isozyme expresses in the grain.By the method of Native-PAGE, it is confirmed that the newly discovered protein hybrid GSX and GSX2, from the crude extract of flowering of flag leaf, are GS isozymes.By the pot experiment in field , simulating field, the crude extract of flowering of flag leaf is studied by the method of SDS-PAGE. It is reported that the GS isozyme subunit exist 87.8KD, 68.6KD, 37KD, 19.05KD, 17.85KD subunit except the previously reported 43.6KD and 38.4KD subunits.By the method of Native-PAGE, the proteins of GS1,GS2,GSX from the jointing stage and the proteins of GS1,GS2,GSX,GSX2 from the period of flowing are recovered respectively, the results indicated that the GS1,GS2,GSX subunits have changed at different growth stages of the wheet .Compared to the expression of GS isozymes in the jointing stage, the GS1 subunits increased the types of 43.6KD subunits and is lack of the 57.4KD and 37KD subunit in the flowering;GS2 subunits increased the 38.4KD subunits and missing the 57.4KD subunits;GSX subunits missing the types of 87.8KD subunits and increased the 37KD subunits.The constitute of GSX2 subunits in the flowing are approaching to the GS1 subunits in the same period.The effects of different N forms on the changement of GS isozyme subunits during the jointing stage of yumai 34 were studied, the result showed that nitrate can promote the content of various types of subunits of GS isozyme from yumai 34. Ammonium salt can facilitate the types of GS isozyme subunits of parts of organizations.Compared to the nitrate and ammonium, the effect of urea on the GS isozymes was relatively lower.The effects of nitrate on the composition and the changements of contents of GS isozymes in the different growth stages of yumai 34 were studied, it was found that the tendency of GS isozymes'types of subunits composition and content during different growth was first increased and then decreased. The GS isozymes were affected greately by different growth stages. In the flowering , the types and contents of GS isozymes were highest .During the different phases of yumai 34, reproductive organs of GS isozymes and content of order were leaves, sheath, section, root. The types of GS isozymes of grain and glume were more unique.By the method of two-dimensional electrophoresis, the whole proteins of flag leaves with the treatment of nitrate of yumai 34 were extracted and studied, the result showed that two kinds of GS isozymes, GS2 and GS2a, were captured and isolated. And 4 kinds of unknown proteins were captured, too. |
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