| Maize is an important food,feed and industrial raw material.The proportion of the cultivated area of maize in China is 25%of the total cultivated area in the world.The yield and quality of maize are of great significance to China's agriculture and economic development.There are many factors affecting the yield and quality of maize,among which fungal disease is one of the most important factor.The maize ear rot is a kind of fungus disease which is common and harmful in the world,the yield and quality of maize were seriously affected and the safety of human and livestock were seriously threatened.The main pathogen of maize ear rot is Fusarium verticillioides.The selection and breeding of disease-resistant varieties is the fundamental method to solve the hazard of F.verticillioides in the maize,which requires a deep understanding of the disease-resistant genetic mechanism and disease-resistant genes.In this study,Two genes related to resistance to F.verticillioides were selected by analysis of genome-wide association study analysis(GWAS),transcriptome and QTL mapping interval of F.verticillioides in maize,were named GRMZM2GGI8133(ZmBGLU)and GRMZM2G154628(ZmPIP2d).Two candidate genes were cloned by homologous cloning,and the vectors were constructed for genetic transformation and functional analysis of candidate genes.The results are as follows:(1)The full length of ZmBGLU and ZmPIP2d were cloned in high resistance maize inbred line BT-1 and high sensitivity inbred line N6.The transient expression vectors of ZmBGLU and ZmPIPZd were constructed and transformed into intraepidermal cells of onion,both of two encoded proteins were located on the cytoplasmic membrane;(2)The silencing vectors of ZmBGLU and ZmPIPZd were constructed through Gateway reaction method,and maize was transformed to obtain T2 generation silencing lines;(3)The silencing lines of ZmBGLU spike-seeds and spike-stalk improved the resistance to F.verticillioides in N6 and BT-1,while the resistance of transgenic maize to F.verticillioides increased in mature grains and seedlings;The silencing lines of ZmPIP2dmature grains improved the sensitivity to F.verticillioides in N6 and BT-1,while the resistance of transgenie maize to F.decreased in spike-seeds,spike-stalk and seedlings.Based on the previous results and the data analysis,two candidate genes related to the resistance of F.verticillioides were selected.The results showed that ZmBGLU and ZmPIP2d could respectively reduce and enhance the resistance of maize to F.verticillioides through the construction of transgenic silenced lines and functional verification of ZmBGLU and ZmPIP2d,which provided the good genetic resource for the cultivation of maize varieties resistant to maize ear rot. |
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