Application Of The Gene OrfH79 In The Creation Of Male Sterile Materials In Wheat

Wheat is an important grain crop in our country and the world grain production plays a pivotal role.In recent years,the yield and quality of wheat increase slowly due to the continuous reduction of cultivated land area and the constant change of environment and climate.In order to meet people’s growing demand for food security and production of high-quality varieties,increasing wheat yield and improving wheat quality will remain important breeding goals,which requires continuous innovation in breeding techniques and germplasm resources.In recent years,the continuous development of gene editing technology has accelerated the pace of functional gene research in wheat,and also provided a new opportunity for the improvement of wheat disease resistance and stress resistance.SPT seed production technology is a two-line seed production technology based on sterile line seed production,it realizes the dual use of sterile line and maintainer line,effectively and conveniently distinguish sterile line and fertile line,sterile line production and hybrid production without transgenic,avoiding the controversy of transgenic food safety issues.The creation of male sterile lines is the key to seed production of sterile lines.In this study,rice mitochondrial cytoplasmic male sterility gene orfH79,driven by maize pollen specific polygalacturonates gene promoter PG47,was connected with mitochondrial signal peptide to form inverters.and the expression vector of wheat Taigu male sterility gene Ms2,driven by PG47,was constructed to convert common wheat Fielder.In order to test whether the specific expression of rice mitochondrial cytoplasmic male sterility gene orfH79 and wheat Taigu nuclear sterility gene Ms2 in anther can achieve wheat male sterility,and obtain new wheat sterility materials,so as to provide theoretical and material basis for hybrid wheat breeding.The main research results are as follows:(1)orfH79 gene expression vector driven by PG47 was constructed,and genetic transformation of common wheat Fielder was carried out,and 7 positive transgenic strains were obtained.(2)A PG47-driven Ms2 gene expression vector was constructed and three positive transgenic strains of wheat Fielder were obtained.Ms2 gene driven by PG47 promoter was not expressed in roots,stems and leaves of wheat,but only in anthers.The anthers of transgenic materials showed obvious characteristics of sterilization.(3)In order to further clarify the expression characteristics of maize PG47 promoter in wheat,we also constructed GUS reporter expression vectors driven by PG47,and carried out genetic transformation of common wheat Fielder.No corresponding transgenic materials have been obtained yet.(4)Tissue specific expression analysis and pollen activity investigation of Ms2 transgenic wheat confirmed that PG47 promoter driven Ms2 expression box could achieve anther specific expression and cause male sterility in wheat.