A novel 10 kDa protein with anti-HIV-1 reverse transcriptase (RT) inhibitory activity was isolated from leaves of Canna indica L. using a combination of native-PAGE and ammonium sulfate precipitation. HIV-1 and RT inhibitory activity was measured using a syncytium forming (deltaTat/Rev) MC99 virus in Tat/Rev transfected 1A2 cell line and ELISA technique, respectively. Edman N-terminal and internal amino acid sequence (using LC-MS-MS) determination revealed the 10 kDa Canna indica L. leaf protein as a putative plastocyanin. This is the first report of a plant plastocyanin with HIV-1 RT inhibitory property.