Propolis has strong antimicrobial activity, and is often used as drugs or health products. Twenty nine different propolis from 23 provinces of China were selected and studied in this paper, which were further classified into five groups according to the climate zone where collected. The size of inhibition zone and minimum inhibitory concentration (MIC) of 9 different strains (Staphylococcus aureus, Bacillus subtilis, Aerobacter aerogenes, Escherichia coli, Proteus, Penicillium, Aspergillus, Mucor, Rhizopus) by the ethanol extract of different propolis (EEP) were measured. The results showed that EEP had good antimicrobial activity against Gram-positive bacteria, Gram-negative bacteria and fungi. The average inhibition zone diameter of Gram-positive bacteria, Gram-negative bacteria and fungi were 988mm±147mm, 875mm±156mm and 1001mm±065mm, respectively. EEP exhibited the highest antimicrobial activity against fungi, while the lowest activity against Gram-negative bacteria. The results indicated that the antimicrobial activity of EEP had no significant correlation with their climate zone.