The Analysis Of Nitrogenous Compounds And Genes Expression Of GS And GLba Among Different Protein Content Materials Of Glycine Max, Glycine Max Of High Protein Content And Glycine Soja Of High Protein Content

Nowadays, soybean is ideal protein sources. Protein content is an important indicator forquality of soybean, it is also one of the main factors which determine the economic value for thecrop. Soybean was planted widely in northeast China and improving protein content of soybeanwill be very important to increase economic efficiency and farmers’ income. In this experiment,in order to understang the genetic characteristics of the formation of protein of G.soja with highprotein content, the changes of nitrogen metabolism-related physiological indicators, GS genesexpression quantity of roots, stems, leaves and nodules and Glba genes expression quantity ofnodules in the whole growth stage were studied using3different protein content materials ofGlycine max （GM）, Glycine soja with high protein content （GSHP） and Glycine max with highprotein content （GMHP） which will also provide an important theoretical guidance toformulating strategies of cultivation and breeding of soybean.1. Soluble protein, free amino acids and nitrate content and nitrate reductase activity （NRA）and glutamine synthetase activity （GSA） of the roots, stems and leaves during the whole growthstage were compared and analyzed among3soybeans with different protein content. TheResults showed that: the leaf soluble protein content of GMHP decreased sharper than that ofGSHP after R3. Leaf NRA of GSHP was higher than that of CMHP during the whole growthperiod. GSA of root, stem and leaf was variation as a single peak curve and R3is the highest ofcurve for3soybeans. GSA of root, stem and leave of GSHP increased obviously at R2-R3stagethen descended slowly after R3and were higher than that of GMHP and GM in later growthperiod.GSA of root was higher than that of stem and leaves, and that of leaves is the lowest fordifferent3soybeans.2. GS and Glba were cloned respectively from nodules of GSHP and GMHP. The Glbagene is578bp which is99%similarity with sequence （NM₀01248999.1） published inGenBank. GSγ1gene is1215bp and98.68%similarity with GenBank sequence （X81700.1）. Thesequence of2genes were similar between GSHP and GMHP. The Prokaryotic expression resultsshowed that the2sequences could be expressed in E. coli and the molecular weight of proteinwere about16KDa and44KD that were same to the reported. The result will contribute toprotein purified, antibodies preparation and the function analysis of proteins.3. The qPCR was carried out for RNA expression quantity of Glba of nodules, GS2andGS1gene family members of roots, stems, leaves and nodules during the whole growth period of3soybeans. The results showed that Glba expression of GSHP was higher than that of GMHPat V3stage but the Glba expression of GMHP nodule was significantly higher than that in GSHP.The expression of each GS member fom root, stem, leaf and nodules of3soybeans was ofobvious characteristics. The same gene had significantly different expression in different organs,GSβ1and GSβ2expressed in the root, stem, leaf and nodules in all periods, however, theexpression of GSβ1was dominant in all4organs. The expression quantity of GS2in leaf wassignificantly higher than that of any other organ. GSγ1expression of nodule increased but theexpressions of GSγ1in roots, stems, leaves were very low. Expressions of GS members werealso significant differences of different protein content of soybean.The expression quantities of GSβ1of high protein content soybean were higher than that ofGM in more growth period. The expression quantity of GS2in leaves and GSγ1in nodulesincreased significantly and from V3to R3they showed the same law, GSHP> GMHP>GM,with the the expression quantities of GSβ1and the total gene expression quantity among3soybeans. The law of leaf and nodules was more obvious than that of other organs and the lawwas coincident with the rule of GSA of three types of soybean Hence, the soybeans of highprotein content could more effectively regulate the expression of every gene of GS throughoutthe growth period than that of GM and obtain high GSA, which is one of physiologicalcharacteristics of high protein content soybean.