The Establishment Of Genetic Transformation System For The Salt Tolerance Transgenic Soybean

In this study,10soybean varieties of Jidou series were used as the experimentmaterial. The appropriate genotypes of transgenic experiments were obtainedthrough a series of research on disinfection method, the comparison on salt toleranceof various varieties, as well as the concentration of6-BA. The references for genetictransformation of salt tolerance gene were provided by PCR and productssequencing for GmGT-2A on27soybean varieties. Finally, based on thecomprehensive analysis of these conditions, suitable for salt tolerance of transgenicsoybean system were established.Bacterial contamination rate, hypocotyl length, radicle length and the radiclenumber of the varieties treated with mercuric chloride and chlorine disinfectantswere compared. The results showed that chlorine disinfection was better thanmercuric chloride disinfection.Statistics on salt tolerance of the10soybean varieties were conducted by settingthe ladder concentration at0mmol/L,50mmol/L,100mmol/L and150mmol/L forsalt stress medium. Results indicated salt-tolerant ability of Jidou4and Jidou8wasstronger than the others;100mmol/L of salt stress concentration was a suitableconcentration to screen transgenic soybeans of salt tolerance.0.5mg/L6-BA was better for the regeneration of Jidou varieties. Jidou5, Jidou7and Jidou9were relatively high bud regeneration rate and moderate rootregeneration rate; they were suitable to do the subsequent genetic transformation.In the transformation of GmGT-2A genetic experiment, germination mediumadded0.5mg/L6-BA played a promotion role of regeneration; Km had obviousinhibitory effect on bud inducement; Plant regeneration also existed differencebetween Jidou9and Jidou7, and the former was better than the latter.Specific PCR results of target gene GmGT-2A showed that SNPs were found indifferent soybean varieties. ShanxiA301had thirteen mutations, while Jidou8hadfive mutations. All tested varieties had five common mutations in DNA sequence ofGmGT-2A, they were respectively three GC into AT in the location of866,910,992,AT into CG in the location of982, and CG into AT in the location of1001.