Penicillium is one of the most important postharvest pathogens of citrus fruits worldwide. It induces blue or green mold disease, a decay that can lead to significant economic losses during storage. Based on internal transcribed spacer (ITS) sequences, seven Penicillium species and one closely related Talaromyces variabilis were identified from 30 rotten samples of citrus fruits marketed in Qena. Penicillium expansum was the most common species, recovered from 16.7% of the samples, followed by P. chrysogenum (10%) and P. polonicum (10%). Sixteen isolates were tested through inoculation on healthy citrus fruits; the data exhibited that 68.7% of isolates were highly virulent. A "Specific Gene Random Primer Polymerase Chain Reaction (SGRP-PCR)" marker technique indicated that the genetic similarity among P. expasum ranged from 49.4 to 85.7%, and a relatively correlation was found between SGRP band profile and species origin. Patulin was detected in 40% of P. expansum isolates. This study provided a useful molecular approach to identify different Penicillium species by sequencing ITS region, focus on the pathogenicity, compare between P. expansum isolates and their ability in patulin production.