| Weeds in soybean field seriously limit the increase of yield and quality of soybean.The application of glyphosate in field is limited due to its wide species’extinction effect.Biotech genetically modified crops resistant to glyphosate allow farmers to use glyphosate to control weeds.In order to obtain GM soybeans,we introduced the G10-EPSPS gene into cultivar Zhongdou 32 through Agrobacterium mediated method in this study.Then we made molecular confirmation and field glyphosate-resistant trial test,transcripts sequencing,also assessed the biodiversity influence and food safety of the transgenic soybean progeny,we made the following conclusions:1.Southern blot analysis confirmed that G10-EPSPS gene has successfully integrated into the soybean plant genome with one copy.Western blot analysis and Elisa result showed that the G10-EPSPS protein expressed in all of the testing tissues,such as root,stem,leaves,flowers,pod shell and seeds.The foreign protein content in leaves is higher than those of all tested tissues,and the content in leaves in pod period is higher than that in seedling period.The foreign protein content in seed is the lowest.There is no statistically difference of G10-EPSPS protein in leaves and seeds in different（T7、T8、T9）generation,and the foreign protein expressed stable in different generations.The ratio of resistance to sensitive plants（number of survival plant to died plant）of T1generation conformed 3:1.2.Based on the Illumina-sequencing platform,93425 transcripts（read length 2×150 bp）were obtained by assembly and splicing.The transcripts number by GO annotations were 14564,the number of annotated genes was 256.The number of transcriptomes annotated by COG is 7263,and the number of genes is 20.The number of transcriptomes annotated by KEGG is 9401,and the number of genes is249,genes are related to cellular processes,environmental information processing,genetic information processing,disease,metabolism and other pathways.According to the glyphosate resistance mechanism,15differentially expressed genes related to the glyphosate metabolic pathway were selected for expression,differential significance and expression regulation study.It was found that DAHPS1,DHQS and DHD-SDH gene in the metabolic pathway of sikimic acid were more expressed in glyphosate-resistant soybean than in the receptor control group,they are the key genes involved in the resistance of transgenic soybean to glyphosate.3.The protein,oil and other 47 nutrients of transgenic soybean were analyzed,and only 2 of them（crude fiber 7.91±0.27g/100g,potassium 1.67±0.02mg/kg）were statistically different from the recipient soybean,and difference between transgenic soybean and receptor lies in the range of the difference variation of conventional soybean.4.Field arthropod investigation and analysis showed that there were significant differences in few indexes among three treatments of transgenic soybean sprayed with glyphosate,transgenic soybean without glyphosate sprayed and recipient soybean without glyphosate sprayed.Weed investigation and analysis showed that there were few significant differences between weed indexes and receptors of transgenic soybean.Most dominant insects and weeds showed no significant difference in distribution and quantity among three treatments.The overall analysis showed that transgenic soybean had no significant effect on the population diversity of arthropods and weeds in the field.5.The results showed that there was no significant difference in seed granulation,seedling yield,and the plants from mature seed generation in field between the genetically modified soybean,receptor and local varieties.The analysis showed that the transgenic soybean had no survival competitive advantage and no weed risk. |
