Abstract: In the paper, the suitable condition for separating and analysing chromosomes DNA of Pyricularia grisea were researched by pulsed field gel electrophoresis The results showed that proteinase E or reagent proteinase made in China can replace proteinase K In a certain electric field installation, the separated effect related to voltage, pulse time, electrophoresis time and temperature of electrophoresis system, among which voltage and pulse time were the main factor It was tested that the effective pulse time 0.5-110 sec was the electrophoresis conditions of the fungus chromosomes DNA.