Abstract: A method for producing maize germplasm with low mycotoxin content and high starch content using gene editing technology, and applications thereof. The method comprises using CRISPR/Cas9 multi-gene editing technology and genetic transformation, wherein, during genetic transformation, an infection medium contains one or more of betaine and calcium chloride. Based on existing studies of functional genes related to starch synthesis and ear rot, the present invention applications high-throughput gene editing to efficiently and randomly aggregate numerous advantageous loci. By adding appropriate concentrations of betaine and calcium chloride during genetic transformation, a higher number of loci are edited in a single transgenic event with better efficiency, thereby effectively increasing diversity and producing more superior locus combination patterns.