Creation Of New Herbicide-tolerant Transgenic Soybean Material With AM79-EPSPS

Compared with the development situation and scientific research situation of soybean industry at home and abroad,it can be seen that cultivating herbicide-tolerant gm soybean and expanding the planting area of gm soybean have become an effective way to achieve the balance between supply and demand of soybean and ensure food security in ChinaGlyphosate as a highly effective and low-toxic broad-spectrum herbicide,has always been favored.AM79-EPSPS Protein not only has a high glyphosate tolerance,but also maintains a strong affinity with PEP,which can give plants resistance to herbicide glyphosate,making it an excellent choice for cultivating glyphosate-resistant transgenic crops.In this study,Soybean Tianlong NO.1 was used as the material to explore and establish an efficient conversion system of AM79-EPSPS herbicide-tolerant soybean from several factors,such as chlorine fumigation time of soybean seed,AS concentration used during infection,the glyphosate concentration of selection agent,the time when IBA was dipped in the rooting process.The results show that the best change was achieved when the chlorine fumigation time of soybean seeds was 6-8 hours,the AS concentration was 200 ?M/L during the infection,the glyphosate concentration of selection agent was 100 ?M/L,and 30 seconds was taken to dip in IBA in the rooting process.According to the transformation system,a batch of transformation was carried out and we got six positive seedlings,The two plants with the highest expression levels,A6 and A8,were screened by ELISA.For these two plants with the highest expression levels,The expression of AM79-EPSPS in different tissues of their offspring at different stages was confirmed by RT-PCR,and the normal expression of exogenous target protein in their offspring was proved by Western blot.Furthermore,Southern blot confirmed that the target gene AM79-EPSPS were inserted into the genome of A6 and A8 in a single copy,and the integration was stable among different generations.This study also analyzed the characteristics of AM79-EPSPS Protein,which has no sequence similarity with known allergens,toxic proteins or anti-nutrition factors that are toxic to humans and mammals,and it can be speculated that AM79-EPSPS Protein,does not have potential sensitization,toxicity and antinutritional effects.