We studied the material collected from agricultural land. Four Ascomycota fungi were isolated from the soil sample. The preliminary identification based on morphological characters using taxonomic keys was done only up to the genus level, due to overlapping characters, making identification difficult. Therefore the Internal transcribed spacer (ITS) region also called as fungal DNA barcode was sequenced. The obtained results were compared with GenBank, the NCBI database, and identification was made up to species level based on the similarity search result. The phylogenetic relationship among the species and between species was studied using a neighbor-joining tree. The genetic distance between the study fungi was computed through the maximum composite likelihood model. These sequences were deposited in GenBank, NCBI, and accession numbers were allotted by GenBank to the submitted sequences. Based on the morphological characters and the DNA sequence analysis results, the isolated fungus was identified as Aspergillus terrus and Aspergillus flavus.