The complete nucleotide sequence of the gene 4 of rice yellow stunt rhabdovirus (RYSV) was determined from cDNAs corresponding to the viral genomic RNA. Gene 4 is 913 nucleotides (nt) long, comprising a 17-nt untranslated 5' region, a 786-nt open reading frame encoding a polypeptide with a molecular mass of 29,125 Da, and a 110-nt untranslated 3' region. Western blot analysis of the RYSV proteins using the antiserum raised against the protein expressed from the cloned gene in Escherichia coli indicates that gene 4 encodes the M protein of RYSV. Comparisons of the deduced amino acid sequence of the M protein of RYSV with those of other rhabdoviruses revealed no significant homologies. However, it shared a similar basic property and a similar distribution of charges with the other rhabdovirus matrix proteins and showed a relatively closer relationship to the sonchus yellow net virus (SYNV) M1 protein.