Evaluation And Gene Cloning Of Glyphosate Resistant Wild Soybean Germplasm In Hebei Province

As a close wild species of cultivated soybean,wild soybean has abundant germplasm resources.In this experiment,wild soybean resistant to herbicide was obtained by screening different concentrations of glyphosate under different conditions;Select some materials in different habitats to determine physiological indicators,The results showed that the screening effect of 600 times liquid concentration using glyphosate was more obvious under open-air potting conditions in Hebei Province.Real-time quantitative PCR was performed on the selected resistant materials and sensitive materials to analyze the fluorescence expression of EPSPS.The trend of change;the cDNA obtained from the resistant material is subjected to a PCR reaction,and the target fragment is amplified and purified to obtain a resistance gene.It laid the foundation for the cultivation of high-quality herbicide resistant varieties in the future.The specific research results are as follows:1.Germplasm screening.According to the research results of previous years,862 wild soybeans were treated with glyphosate at a concentration of 400 and 600 times in the field.The secondary screening was potted in a greenhouse,potted in open air,and treated with 600 times liquid concentration of glyphosate.Some materials in two habitats were selected to determine physiological indexes,and some glyphosate resistant wild soybean materials were selected.2.Optimization of glyphosate screening methods.The physiological indexes of some materials were measured.The content of photosynthetic pigments in potted plants in greenhouses decreased more than that of open-air potted materials,which was seriously damaged by plants.The cost of open-air potted plants was more efficient.Therefore,the use of glyphosate for 600-fold liquid concentration screening in the open-air potting conditions in Hebei Province is more obvious.3.The analysis of the difference in expression of the materials showed that the EPSPS of wild soybean materials were expressed under glyphosate stress,and therelative expression of fluorescence showed an up-and-down and then rising trend within 0~48 h.The resistant material table can respond quickly to glyphosate stress,and EPSPS is expressed in a large amount,However,sensitive materials cann?t form a large amount of epsp synthetase,showing severe glyphosate phytotoxicity.4.The complete open reading frame CDS sequence of glyphosate resistant material 2010 Edition 17 EPSPS gene was cloned.The full-length cDNA was 1582 bp.The DNA sequence and amino acid sequence were different from the registered information...