Abstract: The karyotype analysis of fungi refers to the analysis of the morphological characteristics and characterization of the chromosomes of fungal cells, including the study of the number and length of chromosomes and the position of centromeres, etc., which can provide an important basis for the classification, genetic evolution and chromosome variation of fungi. The common methods of fungal karyotype analysis, its principle and sample preparation were introduced in this paper, and the advantages and disadvantages of each method were compared, so as to provide reference for the selection and improvement of methods for the study of fungal karyotype.