Alfalfa mosaic virus (AlMV) is one of the most conspicuous alfalfa viruses in the world. In order to identify the distribution, host range and molecular characterization of Alfalfa mosaic virus in Kerman province, 514 alfalfa samples showing mosaic, Calico, mottle and malformation symptoms as well as 66 potato samples showing calico symptoms were collected from alfalfa and potato fields in Kerman, Jiroft, Kahnooj, Shahrbabak, Baft, Sirjan and Bardsir regions, Kerman Province. Infection of samples was analyzed through, enzyme-linked immunosorbent assay. In host range studies, AlMV isolates induced different symptoms on several varieties of Nicotiana tabacum L. From among all the Iranian AlMV isolates, only two, namely: Ke. Si. Al, and Ke. Ba. Al caused different viral symptoms including enation and necrotic local lesions on Datura maxima, respectively. As Based on geographical location and biological properties, four isolates were selected for molecular investigation. The whole Coat Protein (CP) gene from four AlMV isolates (three alfalfa and one potato isolate) were PCR-amplified, cloned, sequenced and compared with the counterpart sequences available in the GeneBank. The CP gene of four Iranian isolates of AlMV showed nucleotide and amino acid sequence identities in the ranges between 92.9-98.3 and 91.9-100%, respectively. In phylogenetic analysis, all the four Iranian AlMV isolates were clustered in the Italian group. This is the first report on biology and CP sequence analysis of AlMV isolates from Kerman province in Iran.