Maize is the most important cereal crop in the world after rice and wheat. Generally, Maize known as Queen of Cereals, because it has the highest genetic yield potential among cereals. It is nutrition rich cereal crop. But a large number of fungi can attack and release mycotoxins, causing ear rots. This leads to affect the productivity and marketability of Maize. The isolation and identification of fungi from mature kernels of Maize and soil of rhizosphere region of maize plant was demonstrated. Infected maize kernels and soil samples of maize cultivars P3501 and P3546 were collected and inoculated on modified Czapek-Dox Agar (CZA) medium, Sabouraud Dextrose Agar (SDA) medium and Potato Dextrose Agar (PDA) medium. In modified Czapek-Dox Agar (MCZA) C3H7MgO6P and K2SO4 replaced the MgSO4 and KH2PO4 of the original. This MCZA medium prevents the precipitation of C3H7MgO6P. For isolation of fungi from infected maize seeds and rhizosphere soil samples, serial dilutions of 10-3, 10-4 and 10-5 have taken and inoculated by pour plate and spread plate methods. The plates were incubated at room temperature (35oC±2) for 7days. Fungal colonies were observed and identified under microscope with 40X magnification. Identified colonies again sub-cultured on same MCZA medium, SDA medium and PDA medium and incubated at room temperature (35oC±2) for 10days. The obtained fungal colonies were observed and conidial heads were identified under 40X microscope.