Jowar (Sorghum bicolorL.) plant belongs to family poaceaeis cultivated in warm climates worldwide and it is inhabitant to the tropics and subtropics region. The present investigation was studied physicochemical characterization and rhizosphere soil mycoflora in the field of jowar charcoal rot caused by Macrophomina phaseolina. Mycoflora of infected field of jowar, need to improve the knowledge of diversity of fungi associated with infected rhizosphere soil. Physico-chemical analyses of infected rhizosphere soil were studied by standard soil testing methods andfungal population dynamicsby using soil dilution technique. Sixteen physicochemical parameters were analyzed. It was foundthat,soil has alkaline pH but EC, Ca, Na, S and Mo contents were found least,whereas OC, P and Kwere high in infected soil as compared to standard range. Due to rots infection, chemical contents of soilalso changed. Eleven samples of soil were analyzedduring January-March, 2015 and 2016. In all tested soilsamples, 13 genera and 20 species of fungi were detected. Aspergillus, Mucor, Rhizopus, Penicillium, Fusarium specieswere found dominant in all tested study sites followed by Trichoderma, Monilia, Pythium, Phytophthora, Humicolaetc. Total number of fungal species colony wasfound dominant in Sangola (S) and Mohal (Moh) sites.Percentage of frequency and % of contribution was found more in Aspergillus nigerand Fusarium oxysporum.